Comparative Parasitology 68(2) 2001 - Peru State College
Comparative Parasitology 68(2) 2001 - Peru State College
Comparative Parasitology 68(2) 2001 - Peru State College
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Volume <strong>68</strong> July <strong>2001</strong> Number 2<br />
Formerly the<br />
Journal of the Helminthological Society of Washington<br />
A semiannual journal of research devoted to<br />
Helminthology and all branches of <strong>Parasitology</strong><br />
CONTENTS<br />
CRISCIONE, C. D., AND W. F. FONT. Development and Specificity of Oochoristica<br />
javaensis (Eucestoda: Cyclophyllidea: Anoplocephalidae: Linstowiinae) . 149<br />
CRISCIONE, C. D., AND W. F. FONT. Artifactual and Natural Variation of Oochoristica<br />
javaensis: Statistical Evaluation of In Situ Fixation 156<br />
BOLEK, M. G., AND J. R. COGGTNS. Seasonal Occurrence and Community Structure of<br />
Helminth Parasites in Green Frogs, Rana damitans melanota, from Southeastern<br />
Wisconsin, U. S.A „ 164<br />
FORRESTER, D. J., G. W. FOSTER, AND J. E. THUL. Blood Parasites of the Ring-Necked<br />
Duck (Aythya collaris) on Its Wintering Range in Florida, U.S.A 173<br />
CARKENO, R. A., L. A. DURDEN, D. R. BROOKS, A. ABRAMS, AND E. P. HOBERG.<br />
Parelaphostrongylus tennis (Nematoda: Protostrongylidae) and Other Parasites of<br />
White-Tailed Deer (Odocoileus virginianus) in Costa Rica 177<br />
JOY, J. E., AND R. B. TUCKER. Cepedietta michiganensis (Protozoa) and Batracholandros<br />
magnavulvaris (Nematoda) from Plethodontid Salamanders in West Virginia,<br />
U.S.A x 185<br />
AGUIRRE-MACEDO, M. L., T. SCHOLZ, D. GoNzALEZ-SoLfs, V. M. VIDAL-MARTINEZ, P.<br />
POSEL, G. ARJANO-TORRES, S. DUMAILO, AND E. SIU-ESTRADA. Some Adult<br />
Endohelminths Parasitizing Freshwater Fishes from the Atlantic Drainages of<br />
Nicaragua .. 190<br />
SALGADO-MALDONADO, G., G. CABANAS-CARRANZA, J. M. CASPETA-MANDUJANO, E.<br />
SOTO-GALERA, E. MAYEN-PENA, D. BRAILOVSKY, AND R. BAEZ-VALE. Helminth<br />
Parasites of Freshwater Fishes of the Balsas River Drainage Basin of Southwestern<br />
Mexico 196<br />
SALGADO-MALDONADO, G., G. CABANAS-CARRANZA, E. SOTO-GALERA, J. M. CASPETA-<br />
MANDUJANO, R. G. MORENO-NAVARRETE, P. SANCHEZ-NAVA, AND R. AGUILAR-<br />
AGUILAR. A Checklist of Helminth Parasites of Freshwater Fishes from the<br />
Lerma- Santiago River Basin, Mexico . 204<br />
CURRAN, S. S., R. M. OVERSTREET, T. T. DANG, AND T. L. NGUYEN. Singhiatrema vietnamensis<br />
sp. n. (Digenea: Ommatobrephidae) and Szidatia taiwanensis (Fischthal<br />
and Kuntz, 1975) comb. n. (Digenea: Cyathocotylidae) from Colubrid Snakes in<br />
Vietnam . ; 219<br />
KUZMIN, Y., V V TKACH, AND S. D. SNYDER. Rhabdias ambystomae sp. n. (Nematoda:<br />
Rhabdiasidae) from the North American Spotted Salamander Ambystoma maculaturn<br />
(Amphibia: Ambystomatidae) 228<br />
(Continued on Outside Back Cover)<br />
Copyright © 2011, The Helminthological Society of Washington
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Executive Committee Members-at-Large: ALLEN L. RICHARDS, <strong>2001</strong><br />
BENJAMIN M. ROSENTHAL, <strong>2001</strong><br />
PETER J. HOTEZ, 2002<br />
RONALD C. NEAFIE, 2002<br />
Immediate Past President: ERIC P. HOBERG<br />
COMPARATIVE PARASITOLOGY<br />
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EDITORIAL BOARD<br />
<strong>2001</strong><br />
WALTER A BOEGER<br />
WILLIAM F. FONT<br />
DONALD FORRESTER<br />
J. RALPH LICHTENFELS<br />
JOHN S. MACKIEWICZ<br />
BRENT NICKOL<br />
WILLIS A. REID, JR. AND JANET W. REID, EDITORS<br />
2002<br />
DANIEL R. BROOKS<br />
HIDEO HASEGAWA<br />
SHERMAN S. HENDRIX<br />
JAMES E. JOY<br />
DAVID MARCOGLIESE<br />
DANTE S. ZARLENGA<br />
© The Helminthological Society of Washington <strong>2001</strong><br />
ISSN 1525-2647<br />
2003<br />
ROY C. ANDERSON<br />
DIANE P. BARTON<br />
LYNN K. CARTA<br />
RALPH P. ECKERLIN<br />
FUAD M. NAHHAS<br />
DANNY B. PENCE<br />
This paper meets the requirements of ANSI/NISO Z39.48-1992 (Permanence of Paper).<br />
Copyright © 2011, The Helminthological Society of Washington
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 149-155<br />
Development and Specificity of Oochoristica javaensis (Eucestoda:<br />
Cyclophyllidea: Anoplocephalidae: Linstowiinae)<br />
CHARLES D. CRISCIONE' AND WILLIAM F. FONT2<br />
Department of Biological Sciences, Southeastern Louisiana University, Hammond, Louisiana 70402, U.S.A.<br />
(2 e-mail: wffont@selu.edu)<br />
ABSTRACT: Because assumptions of strict host specificity and geographic isolation apparently have been used<br />
as criteria in determining species of Oochoristica, studies were conducted to address the effects that these<br />
assumptions could have on resolving the taxonomy of Oochoristica. Experimental infections of native fence<br />
lizards, Sceloporus undulatus imdulatus, and Indo-Pacific geckos, Hemidactylus garnotii, demonstrated that the<br />
exotic tapeworm Oochoristica javaensis lacked host specificity. In addition, tapeworms with gravid proglottids,<br />
a stage of development that has not been previously reported for any species of Oochoristica, were obtained<br />
from both hosts. Evidence against the assumption of geographic isolation stems from the fact that lizard species<br />
known to harbor Oochoristica spp. have been introduced beyond their native ranges, and in some cases, these<br />
introductions predate the species descriptions. Lack of support for either assumption indicates a need for more<br />
rigorous analyses and experimentation to determine species of Oochoristica.<br />
KEY WORDS: Oochoristica javaensis, Cestoda, Hemidactylus turcicus, Hemidactylus garnotii, Sceloporus undulatus<br />
undulatus, Mediterranean geckos, Indo-Pacific geckos, fence lizards, development, host specificity, biogeography,<br />
taxonomy.<br />
Approximately 80 species have been described<br />
in the cosmopolitan genus Oochoristica<br />
Liihe, 1898 (Bursey and Goldberg, 1996a, b;<br />
Bursey et al., 1996, 1997; Brooks et al., 1999).<br />
These anoplocephalid cestodes predominantly<br />
parasitize lizards, but also snakes, turtles, and a<br />
few marsupials (Schmidt, 1986; Beveridge,<br />
1994). Development has been examined for<br />
Oochoristica vacuolata Hickman, 1954, Oochoristica<br />
osheroffi Meggitt, 1934, and Oochoristica<br />
anolis Hardwood, 1932 (Hickman, 1963;<br />
Widmer and Olsen, 1967; Conn, 1985). Although<br />
larval and adult development has been<br />
examined for 3 species of Oochoristica, only<br />
Conn (1985) tried to determine host specificity<br />
experimentally. In his experiments, however, he<br />
was unable to infect wall lizards, Podarcis muralis<br />
(Laurenti, 17<strong>68</strong>) and mice, Mm musculus<br />
Linnaeus, 1758. Curiously, no other attempts<br />
have been made to determine the specificity of<br />
a species of Oochoristica. This is unfortunate in<br />
that, as Brooks et al. (1999) pointed out, one of<br />
the major criteria used in resolving the taxonomy<br />
of Oochoristica has been the assumption of<br />
a high degree of host specificity exhibited by<br />
species in this genus. They also mentioned re-<br />
1 Corresponding author. Current address: Department<br />
of Zoology, 3029 Cordley Hall, Oregon <strong>State</strong><br />
University, Corvallis, Oregon 97331, U.S.A. (e-mail:<br />
crischar@bcc.orst.edu).<br />
149<br />
striction to particular geographic regions as a<br />
criterion that has ostensibly been used in the past<br />
to identify species of Oochoristica.<br />
In a survey of helminths of the Mediterranean<br />
gecko, Hemidactylus turcicus (Linnaeus, 1758),<br />
from Louisiana, U.S.A., a species of Oochoristica<br />
was recovered (C. D. Criscione, unpublished<br />
data); however, there were difficulties in<br />
identifying this species. These problems were<br />
associated in part with the assumptions of strict<br />
host specificity and geographic isolation. It became<br />
apparent that in addition to the lack of<br />
specificity experiments, introduced and native<br />
host distributions were often ignored when identifying<br />
species of Oochoristica. Neither assumption<br />
has been properly addressed, and in order<br />
to validate the identification of any species of<br />
Oochoristica, these assumptions should be tested.<br />
In light of these problems with the taxonomy<br />
of Oochoristica, the primary objective of this<br />
study was to examine the development of Oochoristica<br />
javaensis Kennedy, Killick, and Beverley-Burton,<br />
1982, and to test the assumption<br />
of specificity via experimental infections. In addition,<br />
we comment on the assumption of geographic<br />
isolation.<br />
Materials and Methods<br />
To minimize variation among hosts, gravid proglottids<br />
of O. javaensis were obtained from 15 worms recovered<br />
from the small intestine of a single female<br />
Copyright © 2011, The Helminthological Society of Washington
150 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
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CRISCIONE AND FONT—DEVELOPMENT OF OOCHORISTICA JAVAENSIS 151<br />
Table 2. Measurements of Oochoristica javaensis obtained from experimentally infected Hemidactylus<br />
garnotii (HEGA), and Sceloporus u. undulatus (SCUN) for day 105 postexposure; measurements in (xm<br />
unless noted otherwise.<br />
Variable<br />
Total<br />
Proglottid number<br />
Neck<br />
Scolex<br />
Sucker<br />
Immature proglottid<br />
Genital pore position||<br />
Mature proglottid<br />
Cirrus sac<br />
Ovary<br />
Vitellaria<br />
Testis<br />
Testes number<br />
Gravid proglottid<br />
Oncosphere<br />
Hook<br />
L± (mm)<br />
W<br />
L (mm)<br />
W<br />
L<br />
W<br />
L<br />
W<br />
L<br />
W<br />
L<br />
W<br />
L<br />
W<br />
L<br />
W<br />
L<br />
W<br />
L<br />
W<br />
L (mm)<br />
W<br />
L<br />
L<br />
Sample<br />
size*<br />
5<br />
5<br />
5<br />
5<br />
5<br />
5<br />
5<br />
5<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
HEGA<br />
n = 5f<br />
SCUN<br />
Sample<br />
size n = 1<br />
61.6-86.9 (67.7 ± 48.3)§ 1 54.5<br />
109-144 (128 ± 5.7)<br />
1 140<br />
166-229 (191 ± 10.7) 1 221<br />
1.16-1.79 (1.52 ± 0.12) 1 1.12<br />
90-191 (152 ± 17.7) 1 218<br />
78-277 (140 ± 36.3) 1 164<br />
35-90 (61.8 ± 9.5) 1 82<br />
47-82 (66.4 ± 6.4) 1 105<br />
482-561 (520 ± 6.6)<br />
3 379-403 (390 ± 7.1)<br />
300-387 (348 ± 7)<br />
3 427-466 (443 ± 11.9)<br />
0.24-0.28 (0.27 ± 0.004) 3 0.24-0.28 (0.26 ± 0.01)<br />
593-695 (643 ± 9.1) 3 403-411 (406 ± 2.7)<br />
616-790 (712 ± 12.4) 3 648-695 (679 ± 15.7)<br />
43-51 (46.7 ± 0.47) 3 47-51 (49.7 ± 1.3)<br />
137-164 (146 ± 1.99) 3 109-121 (117 ± 4.0)<br />
265-351 (314 ± 6.0)<br />
3 1<strong>68</strong>-211 (91 ± 12.6)<br />
195-265 (230 ± 5.1)<br />
3 133-187 (159 ± 15.7)<br />
137-191 (169 ± 4.5) 3 86-105 (96.3 ± 5.6)<br />
82-144 (118 ± 4.3) 3 82-101 (89.7 ±5.8)<br />
39_47 (43.5 ± 0.77) 3 35-39 (36.3 ± 1.3)<br />
35-55 (46.2 ± 1.3) 3 39-43(41.7 ± 1.3)<br />
23-35 (30.8 ± 0.78) 3 21-30 (26.3 ± 2.7)<br />
571-749 (629 ± 12.6) 3 433-473 (453 ± 11.6)<br />
1.50-2.28 (1.92 ± 0.05) 3 1.48-1.52 (1.49 ± 0.01)<br />
20-34 (25.9 ± 1.04) 3 22 (22 ± 0.0)<br />
16-28 (21.5 ± 0.79) 3 18-20(19.3 ± 0.67)<br />
10-12 (11.6 ± 0.21) 3 12 (12 ± 0.0)<br />
* Sample size refers to the total number of each character that was measured.<br />
t Number of tapeworms measured.<br />
± L = length, W = width.<br />
§ Range followed by mean ± 1 SE in parentheses.<br />
|| Genital pore position was calculated as a ratio of the position along the length of the mature proglottid from the anterior end<br />
(length to the center of the genital pore -^ length of proglottid).<br />
mediate host for the genus Oochoristica. Oochoristica<br />
javaensis became established in 7 individuals<br />
of the experimental definitive hosts.<br />
Successful infection occurred in only 1 of 16 H.<br />
turciciis; this Mediterranean gecko was examined<br />
on day 1 PE and had an intensity of 4.<br />
Oochoristica javaensis was recovered from 3 of<br />
10 H. garnotii on days 7, 28, and 105 PE and<br />
had intensities of 1, 7, and 6, respectively. Three<br />
of 5 S. u. undulatus were infected with 2, 3, and<br />
1 tapeworms on days 10, 30, and 105 PE, respectively.<br />
The 10 A. carolinensis and 5 R.<br />
sphenocephala were negative for infections.<br />
Measurements for specimens from experimental<br />
infections are in Tables 1 and 2.<br />
Proglottid formation did not occur prior to<br />
day 28 PE (Figs. 1 and 2). Terminal proglottids<br />
of tapeworms recovered on day 28 PE from H.<br />
garnotii had developing ovaries, testes, vitellaria,<br />
and cirrus sacs (Fig. 3). A median terminal<br />
excretory pore was present in each terminal proglottid,<br />
and in 1 worm there was a developed<br />
genital atrium. Terminal proglottid length ranged<br />
from 514 to 822 (mean = 659, SE = 38.7, n =<br />
7) and width from 174 to 277 (mean = 216, SE<br />
= 12.4, n — 7). Two of the worms from S. u.<br />
undulatus examined on day 30 PE also had developing<br />
reproductive organs (Fig. 4). One measured<br />
442 X 158 (L X W); the other had been<br />
torn. The third tapeworm recovered from day 30<br />
PE had been damaged in the mounting process;<br />
however, prior examination had shown that no<br />
more than 15 proglottids had formed, sexual primoridia<br />
had just begun to develop, and total<br />
Copyright © 2011, The Helminthological Society of Washington
152 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Copyright © 2011, The Helminthological Society of Washington
length did not exceed 1.2 mm. By day 105 PE,<br />
development of O. javaensis progressed to strobilas<br />
with gravid proglottids in both H. garnotii<br />
(Fig. 5) and 5. u. undulatus (Fig. 6). Although<br />
it was possible that tapeworms from day 105 PE<br />
were natural infections because experimental definitive<br />
hosts were not laboratory-raised, prior<br />
fecal examinations were negative for all hosts<br />
used in experiments.<br />
Discussion<br />
Development of Oochoristica javaensis<br />
Prior to the present study, the most developed<br />
stage experimentally obtained for a species of<br />
Oochoristica was a terminal mature proglottid<br />
(Conn, 1985). Our experimental infections with<br />
O. javaensis were successful in obtaining gravid<br />
specimens. Susceptible definitive hosts for O. javaensis<br />
included H. turcicus, H. garnotii, and 5.<br />
u. undulatus; however, the fact that only 1 of 16<br />
control hosts, H. turcicus, became infected suggests<br />
that exposure techniques may have been<br />
flawed. Possible problems may have been the<br />
temperature at which experimental hosts were<br />
housed or the inoculation method. When dealing<br />
with small hosts, the stomach tube may not be<br />
the best method, and another, such as placing<br />
metacestodes in gel capsules, may prove to be<br />
more efficient. Despite the scarcity of infections,<br />
however, the developing worms obtained from<br />
H. garnotii and S. u. undulatus indicated a lack<br />
of specificity for O. javaensis.<br />
On day 105 PE, Conn (1985) recovered a single<br />
specimen of O. anolis from A. carolinensis<br />
that had mature proglottids with fully formed<br />
male and female reproductive systems. The terminal<br />
proglottid, however, still had a median excretory<br />
pore, suggesting that this specimen had<br />
not yet shed any proglottids. Specimens of O.<br />
javaensis from H. garnotii on day 28 PE also<br />
had median excretory pores in the terminal proglottids.<br />
Although most of the specimens that we<br />
recovered from day 28 PE did not have fully<br />
developed reproductive organs, their stage of development<br />
had greatly surpassed the develop-<br />
CRISCIONE AND FONT—DEVELOPMENT OF OOCHORISTICA JAVAENSIS 153<br />
mental stage of O. anolis from day 28 PE (Conn,<br />
1985). Oochoristica anolis from green anoles on<br />
day 28 PE had just begun to form proglottids<br />
with genital anlages and had a maximum total<br />
length of 3.25 mm (Conn, 1985). Widmer and<br />
Olsen (1967) reported immature O. osheroffi<br />
with a maximum total length of 4.14 mm on day<br />
28 PE in the prairie rattlesnake, Crotalus viridis<br />
Rafinesque, 1818, but the mean total worm<br />
length from our study for day 28 PE was 8.32<br />
mm (Table 1). These comparisons suggest a<br />
more rapid development in the definitive host<br />
for O. javaensis than for O. anolis or O. osheroffi.<br />
However, small sample sizes in our study<br />
and infection techniques differing from previous<br />
life cycle studies of Oochoristica spp. prevented<br />
definitive comparison of developmental patterns<br />
among species. Likewise, the low number of infections<br />
precluded examination of host-induced<br />
variation for O. javaensis. Although development<br />
in S. u. undulatus appeared slightly slower<br />
than in H. garnotii up to day 30 PE (Table 1),<br />
measurements of gravid specimens from H. garnotii<br />
and S. u. undulatus on day 105 PE (Table<br />
2) may suggest plasticity for some characters.<br />
Host specificity and geographic isolation<br />
Results from our study experimentally demonstrated<br />
for the first time that a single species<br />
of Oochoristica can infect more than 1 species<br />
of host. Oochoristica javaensis infected hosts<br />
representing 2 unrelated lizard families, Phrynosomatidae<br />
for S. u. undulatus and Gekkonidae<br />
for H. garnotii (Estes et al., 1988; Pough et al.,<br />
1998). It is not known if the ecology of either<br />
S. it. undulatus or H. garnotii would predispose<br />
natural populations of these hosts to the establishment<br />
of O. javaensis. Development in different<br />
hosts demonstrated via our laboratory experiments,<br />
however, raises questions with regard<br />
to the use of host specificity as a taxonomic criterion<br />
for species of Oochoristica. Specificity in<br />
the laboratory and in the field should be examined<br />
for other species of Oochoristica in light of<br />
these results for 2 reasons. First, lack of speci-<br />
Figures 1-6. Development of Oochoristica javaensis from Hemidactylus garnotii (HEGA) and Sceloporus<br />
u. undulatus (SCUN) at different days postexposure. Photomicrographs were taken with differential<br />
interference contrast. Bars = 200 jjim. 1. Day 7 in HEGA. 2. Day 10 in SCUN. 3, 4. Terminal proglottids<br />
from day 28 in HEGA and day 30 in SCUN, respectively. 5, 6. Mature proglottids from day 105 in HEGA<br />
and SCUN, respectively. CS = cirrus sac, GA = genital atrium, O = ovary, T = testis, and V = vitellaria.<br />
Copyright © 2011, The Helminthological Society of Washington
154 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
ficity means that tapeworms of the same species<br />
will be exposed to different environments in different<br />
species of hosts, thus presenting opportunities<br />
for host-induced variation. If variation is<br />
induced, then this may affect the current morphometrically<br />
based taxonomy of species of<br />
Oochoristica. Second, a better understanding of<br />
the degree to which species of Oochoristica can<br />
switch hosts is imperative in light of the conservation<br />
implications associated with introduced<br />
organisms and their parasites (see Barton,<br />
1997).<br />
Introduced lizards will have consequences not<br />
only for conservation, but also for parasite taxonomy.<br />
If in the past, species of Oochoristica<br />
have been transmitted with their exotic lizard<br />
hosts, then current assumptions of biogeographic<br />
isolation may be incorrect. This is not to say that<br />
there was never a biogeographic pattern that paralleled<br />
Oochoristica speciation, but especially<br />
because of anthropogenic effects, species of<br />
Oochoristica may have colonized new areas before<br />
many of them were ever described (see Bursey<br />
et al. [1996] for a list of authority dates).<br />
This possibility exists because records of some<br />
introduced geckos, i.e., H. turcicus in Florida<br />
(Stejneger, 1922) and Hemidactylus mabouia<br />
(Moreau de Jonnes, 1818) in South America and<br />
the Caribbean (Kluge, 1969), predate many<br />
Oochoristica species descriptions.<br />
It is interesting to note that several authors<br />
(Bursey and Goldberg, 1996b; Bursey et al.,<br />
1996; Brooks et al., 1999) listed O. vanzolinii as<br />
a Neotropical species of Oochoristica from Brazil<br />
without regard to the fact that it was described<br />
from the introduced house gecko, H. mabouia<br />
(Rego and Oliveira Rodrigues, 1965). It<br />
has been hypothesized that H. mabouia naturally<br />
colonized the New World via rafting or was<br />
transported during the slave trades over 400 yr<br />
ago (Kluge, 1969). In either case, H. mabouia<br />
colonized the New World from Africa, thus presenting<br />
the opportunity for parasite transport. It<br />
is also interesting to note that in describing O.<br />
javaensis, Kennedy et al. (1982) listed O. vanzolinii<br />
as the species that most resembled their<br />
specimens.<br />
Brooks et al. (1999) stated that both assumptions,<br />
specificity and geographic isolation, were<br />
not evidence for new species, and suggested that<br />
in describing new species many morphological<br />
characters should be provided. We strongly support<br />
their contention; however, the full extent to<br />
Copyright © 2011, The Helminthological Society of Washington<br />
which certain features are plastic is still unknown<br />
for this genus. As indicated by Criscione<br />
and Font (<strong>2001</strong>), proglottid morphology of O.<br />
javaensis exhibited a high degree of plasticity<br />
that may have resulted from a crowding effect<br />
(Read, 1951). Providing more characters may alleviate<br />
some problems, but it will not solve the<br />
underlying difficulties associated with the taxonomy<br />
of Oochoristica. The morphologically<br />
based taxonomy of Oochoristica will only be<br />
validated upon experimentation establishing the<br />
variation of characters within the genus and/or<br />
the use of molecular data.<br />
Acknowledgments<br />
We express our gratitude to Dr. Richard Seigel,<br />
Caroline Kennedy, and Tom Lorenz for collecting<br />
the fence lizards and leopard frogs, and<br />
to Amanda Vincent and Jonathan Willis for their<br />
help with experiments and care of lizards.<br />
Thanks are also extended to Dr. Murray Kennedy<br />
and Dr. Bruce Conn for loaning us specimens<br />
of Oochoristica from their private collections,<br />
and to Judith Price at the Canadian Museum<br />
of Nature (CMNPA) and Pat Pilitt, Dr. Eric<br />
Hoberg, and Dr. J. Ralph Lichtenfels at the<br />
USNPC for their assistance and loan of specimens.<br />
Literature Cited<br />
Barton, D. P. 1997. Introduced animals and their parasites:<br />
the cane toad, Bufo marinus, in Australia.<br />
Australian Journal of Ecology 22:316-324.<br />
Beveridge, I. 1994. Family Anoplocephalidae Cholodkovsky,<br />
1902. Pages 315-366 in L. F. Khalil, A.<br />
Jones, and R. A. Bray, eds. Keys to the Cestode<br />
Parasites of Vertebrates. Commonwealth Agricultural<br />
Bureau, Wallingford, U.K.<br />
Brooks, D. R., G. Perez-Ponce de Leon, and L.<br />
Garcia-Prieto. 1999. Two new species of Oochoristica<br />
Liihe, 1898 (Eucestoda: Cyclophyllidea:<br />
Anoplocephalidae: Linstowiinae) parasitic in<br />
Ctenosaura spp. (Iguanidae) from Costa Rica and<br />
Mexico. Journal of <strong>Parasitology</strong> 85:893-897.<br />
Bursey, C. R., and S. R. Goldberg. 1996a. Oochoristica<br />
macallisteri sp. n. (Cyclophyllidea: Linstowiidae)<br />
from the side-blotched lizard, Ufa stansburiana<br />
(Sauria: Phrynosomatidae), from California,<br />
USA. Folia Parasitologica 43:293-296.<br />
, and . 1996b. Oochoristica maccoyi n.<br />
sp. (Cestoda: Linstowiidae) from Anolis gingivinus<br />
(Sauria: Polychortidae) collected in Anguilla,<br />
Lesser Antilles. Caribbean Journal of Science 32:<br />
390-394.<br />
-, and D. N. Woolery. 1996. Oochoristica<br />
piankai sp. n. (Cestoda: Linstowiidae) and<br />
other helminths of Moloch horridus (Sauria:
Agamidae) from Australia. Journal of the Helminthological<br />
Society of Washington 63:215-221.<br />
-, C. T. McAllister, and P. S. Freed. 1997.<br />
Oochoristica jonnesi sp. n. (Cyclophyllidea: Linstowiidae)<br />
from the house gecko, Hemidactylus<br />
mabouia (Sauria: Gekkonidae), from Cameroon.<br />
Journal of the Helminthological Society of Washington<br />
64:55-58.<br />
Conn, D. B. 1985. Life cycle and postembryonic development<br />
of Oochoristica anolis (Cyclophyllidea:<br />
Linstowiidae). Journal of <strong>Parasitology</strong> 71:10—<br />
16.<br />
Criscione, C. D., and W. F. Font. <strong>2001</strong>. Artifactual<br />
and natural variation of Oochoristica javaensis'.<br />
statistical evaluation of in situ fixation. <strong>Comparative</strong><br />
<strong>Parasitology</strong> <strong>68</strong>:156-163.<br />
Estes, R., K. De Queiroz, and J. A. Gauthier. 1988.<br />
Phylogenetic relationships within Squamata. Pages<br />
119-281 in R. Estes and G. Pregill, eds. Phylogenetic<br />
Relationships of the Lizard Families, Essays<br />
Commemorating Charles L. Camp. Stanford<br />
University Press, Stanford, California, U.S.A.<br />
Hicknian, J. L. 1963. The biology of Oochoristica<br />
vacuolata Hickman (Cestoda). Papers and Proceedings<br />
of the Royal Society of Tasmania 97:81-<br />
104.<br />
Kennedy, M. J., L. M. Killick, and M. Beverley-<br />
Burton. 1982. Oochoristica javaensis n. sp. (Eu-<br />
CRISCIONE AND FONT—DEVELOPMENT OF OOCHORISTICA JAVAENSIS 155<br />
Editors' Acknowledgments<br />
cestoda: Linstowiidae) from Gehyra mutilata and<br />
other gekkonid lizards (Lacertilia: Gekkonidae)<br />
from Java, Indonesia. Canadian Journal of Zoology<br />
60:2459-2463.<br />
Kluge, A. G. 1969. The evolution and geographical<br />
origin of the New World Hemidactylus mabouiabrookii<br />
complex (Gekkonidae, Sauria). Miscellaneous<br />
Publications, Museum of Zoology, University<br />
of Michigan 138:1-78.<br />
Pough, F. H., R. M. Andrews, J. E. Cadle, M. L.<br />
Crump, A. H. Savitzky, and K. D. Wells. 1998.<br />
Herpetology. Prentice-Hall, Upper Saddle River,<br />
New Jersey, U.S.A. 577 pp.<br />
Read, C. P. 1951. The "crowding effect" in tapeworm<br />
infections. Journal of <strong>Parasitology</strong> 37:174-178.<br />
Rego, A. A., and H. Oliveira Rodrigues. 1965. Sobre<br />
duas Oochoristica parasitas de lacertilios (Cestoda,<br />
Cyclophillidea). Revista Brasileira de Biologia<br />
25:59-65.<br />
Schmidt, G. D. 1986. Handbook of Tapeworm Identification.<br />
CRC Press, Boca Raton, Florida, U.S.A.<br />
675 pp.<br />
Stejneger, L. 1922. Two new geckos to the fauna of<br />
the United <strong>State</strong>s. Copeia 1922:56.<br />
Widmer, E. A., and O. W. Olsen. 1967. The life<br />
history of Oochoristica osheroffi Meggitt, 1934<br />
(Cyclophyllidea: Anoplocephalidae). Journal of<br />
<strong>Parasitology</strong> 53:343-349.<br />
We acknowledge, with thanks, the following persons for providing valuable help and insights in reviewing<br />
manuscripts for <strong>Comparative</strong> <strong>Parasitology</strong>: Roy Anderson, Prema Arasu, Carter Atkinson, Scott<br />
Baird, James Baldwin, Diane Barton, Ian Beveridge, Reginald Blaylock, Walter Boeger, Rodney Bray,<br />
Daniel Brooks, Michael Burt, Goran Bylund, Charles Bursey, Ronald Campbell, Melanie Chapman,<br />
Anindo Choudhury, James Coggins, William Coil, David Cone, Bruce Conn, Thomas Cribb, John Cross,<br />
Lawrence Curtis, Murray Dailey, William Davidson, Emmet Dennis, Marie-Claude Durette-Desset, Ralph<br />
Eckerlin, Burton Endo, Alan Fedynich, Stephen Feist, Jacqueline Fernandez, David Fitch, Donald Forrester,<br />
Scott Gardner, Lynda Gibbons, David Gibson, Tim Goater, Stephen Goldberg, Robert Goldstein, Hideo<br />
Hasegawa, Richard Heard, Sherman Hendrix, John Hnida, Eric Hoberg, John Holmes, Patrick Hudson,<br />
David Huffman, Jean-Pierre Hugot, Kym Jacobson, Francisco Jimenez-Ruiz, James Joy, Michael Kent,<br />
Mike Kinsella, Greg Klassen, Ronald Ko, Delane Kritsky, Kevin Lafferty, Murray Lankester, Ralph Lichtenfels,<br />
Eugene Lyons, John Mackiewicz, David Marcogliese, Jean Mariaux, Chris McAllister, Donald<br />
McAlpine, Frantisek Moravec, Janice Moore, Patrick Muzzall, Fuad Nahhas, Robin Overstreet, Raphael<br />
Payne, Thomas Platt, Robert Poulin, Annie Prestwood, Robert Rausch, Amilcar Rego, Mark Rigby, John<br />
Riley, Klaus Rohde, Benjamin Sacks, William Samuel, Gerhard Schad, Thomas Scholz, Maria Sepiilveda,<br />
Scott Seville, Takeshi Shimazu, John Sullivan, Stephen Taft, Dennis Thoney, Tellervo Valtonen, William<br />
Wardle, Patricia Wilber, and Darwin Wittrock.<br />
Copyright © 2011, The Helminthological Society of Washington
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 156-163<br />
Artifactual and Natural Variation of Oochoristica javaensis:<br />
Statistical Evaluation of In Situ Fixation<br />
CHARLES D. CRISCIONE' AND WILLIAM F. FONT2<br />
Department of Biological Sciences, Southeastern Louisiana University, Hammond, Louisiana 70402, U.S.A.<br />
(e-mail: 2wffont@selu.edu)<br />
ABSTRACT: Lack of knowledge of the extent of natural morphological variation can undermine proper taxonomic<br />
decisions. Confounding this problem is artifactual variation that arises from improper fixation techniques. For<br />
the morphologically based taxonomy of the cestode genus Oochoristica, little information exists on the plasticity<br />
of important taxonomic characters. In addition, paratypes of several species of Oochoristica are highly contracted<br />
and contorted. These paratypes were recovered from preserved hosts; thus, the tapeworms were killed and fixed<br />
inside the host (in situ fixation). Experiments demonstrated that in situ fixation of Oochoristica javaensis results<br />
in highly contracted specimens, and statistical comparisons between relaxed and in situ-fixed tapeworms revealed<br />
significant differences for proglottid measurements. Natural variation for the paratypes recovered from preserved<br />
hosts is likely misrepresented by the artificial variation induced by in situ fixation. Lastly, data from natural<br />
infections suggested that proglottid characters of O. javaensis are plastic and may be subject to crowding effects.<br />
KEY WORDS: Oochoristica javaensis, Cestoda, Hemidactylus turcicus, Mediterranean gecko, fixation techniques,<br />
crowding effects, morphological variation, taxonomy.<br />
The taxonomy of the cestode genus Oochoristica<br />
Liihe, 1898, has been based solely on morphology<br />
without knowledge of the extent of natural<br />
intraspecific morphological variation. Parasite<br />
morphological variation may be the result of<br />
genetic determinants, host-induced effects, parasite<br />
intensity effects, or external habitat influences.<br />
Stunkard (1957) and Haley (1962) discussed<br />
the importance of environmental and<br />
host-induced variation for the systematics of helminth<br />
parasites, citing such factors as different<br />
host species, host age, host diet, or infection intensity<br />
as causes of variation. They also emphasized<br />
the need to assess experimentally the stability<br />
of taxonomic characters when identifying<br />
a species.<br />
In addition to the lack of knowledge on intraspecific<br />
variation, natural variation of some species<br />
of Oochoristica may be masked by artificial<br />
morphological variation induced by fixation<br />
techniques. Several paratype specimens of<br />
Oochoristica examined from the U.S. National<br />
Parasite Collection (USNPC), Beltsville, Maryland,<br />
U.S.A. were highly contracted and contorted.<br />
Examination of the respective species descriptions<br />
revealed that these paratypes (listed<br />
below) were obtained from formalin-fixed hosts.<br />
1 Corresponding author. Current address: Department<br />
of Zoology, 3029 Cordley Hall, Oregon <strong>State</strong><br />
University, Corvallis, Oregon 97331, U.S.A. (e-mail:<br />
crischar@bcc.orst.edu).<br />
156<br />
Copyright © 2011, The Helminthological Society of Washington<br />
That is, they were removed from host specimens<br />
deposited in museum collections without regard<br />
to the effects of host fixation on internal parasites.<br />
Bakke (1988) and others have qualitatively<br />
illustrated the distorting effects of improper fixation<br />
techniques on the morphology of soft-bodied<br />
helminths, but comparisons of fixation techniques<br />
have not been tested statistically to examine<br />
for quantitative differences in the measurements<br />
of important taxonomic characters.<br />
The purpose of our report was to provide a<br />
quantitative assessment of the artifactual morphological<br />
variation induced by killing and fixing<br />
tapeworms within a host, i.e., in situ fixation.<br />
In order to address the effects that improper fixation<br />
methods may have on the morphologically<br />
based taxonomy of Oochoristica, statistical<br />
comparisons of in situ-fixed tapeworms to ones<br />
that were collected alive and killed in a relaxed<br />
state were conducted with specimens of Oochoristica<br />
javaensis Kennedy, Killick, and Beverley-Burton,<br />
1982. In addition, we provide data<br />
regarding the effects of intensity on O. javaensis<br />
morphology.<br />
Materials and Methods<br />
Fixation experiments<br />
To mimic lizard fixation techniques used for museum<br />
collections, 15 Mediterranean geckos, Hemidactylus<br />
turcicus (Linnaeus, 1758), were collected from the<br />
campus of Louisiana <strong>State</strong> University (LSU), Baton<br />
Rouge, Louisiana, U.S.A. (30°24.92'N; 91010.81'W),<br />
where they were known to have a high prevalence of
infection with O. javaensis (C. D. Criscione, unpublished<br />
data). Geckos were killed using an overdose of<br />
ether and immediately fixed via subcutaneous, oral<br />
cavity, and body cavity injections of unheated 10%<br />
formalin. Oral cavity injections insured that the tapeworms<br />
were killed immediately. After 6 days in 10%<br />
formalin, geckos were soaked in water for 24 hr to<br />
remove the formalin. Geckos were then transferred to<br />
70% ethanol for storage until dissection 4 days later.<br />
In situ-fixed tapeworms recovered upon necropsy were<br />
stored in 70% ethanol, stained in Semichon's acetocarmine,<br />
dehydrated in ethanol, cleared in xylene, and<br />
mounted in Canada balsam. Comparisons were made<br />
with relaxed tapeworms that were killed with hot water<br />
(90°C) and fixed and stored in alcohol-formalin-acetic<br />
acid solution (AFA). Relaxed worms were obtained in<br />
a helminth survey of//, turcicus from LSU in the summer<br />
of 1998 (C. D. Criscione, unpublished data); staining<br />
and mounting techniques were the same as for the<br />
in situ-fixed specimens.<br />
Quantitative analyses included measurements of mature<br />
proglottids from 5 relaxed and 5 in situ-fixed specimens<br />
of O. javaensis. Three mature proglottids, located<br />
just anterior to the first proglottid displaying evidence<br />
of egg production, were selected from each individual.<br />
Length and width were measured for each<br />
mature proglottid and for the ovary, vitellaria, and 1<br />
testis within each proglottid. One testis was randomly<br />
selected from each proglottid, ovary length was measured<br />
for the ovary lobe opposite the genital atrium,<br />
and ovary width was measured across both lobes. Although<br />
multiple testes are present within a single proglottid,<br />
only 1 was chosen in order to facilitate the use<br />
of appropriate statistical tests. In order to test for in<br />
situ-fixation effects, a nested ANOVA design was used<br />
to control the pseudoreplication of measuring 3 proglottids<br />
from 1 tapeworm. Tapeworms nested within<br />
type of fixation constituted the experimental units, i.e.,<br />
true replicates, with the error term being the proglottids,<br />
i.e., pseudoreplicates, nested within individual<br />
tapeworms. Principal components analysis (PCA) with<br />
Varimax rotation was used as a data reduction technique<br />
and to examine latent relationships among the<br />
variables. A variable was considered to load on a factor<br />
if its correlation to the factor was >|0.5| (Hair et<br />
al., 1999). The resulting factors with their standardized<br />
factor scores were then tested for differences between<br />
relaxed and in situ-fixed tapeworms in the nested design.<br />
Statistical significance was determined at P <<br />
0.05.<br />
Analysis of natural morphological variation<br />
The analysis of intensity effects on the morphology<br />
of O. javaensis included 5 tapeworms from each of 3<br />
Mediterranean geckos that were naturally infected with<br />
15, 28, and 64 tapeworms. Criteria and morphological<br />
characters used for measurements were the same as<br />
those used in the fixation experiments. Experimental<br />
design using factor scores from a PCA with Varimax<br />
rotation was also the same, in that a nested ANOVA<br />
was used to test for intensity effects. A priori contrasts<br />
of 15 versus 28 and 28 versus 64 were computed. In<br />
order to conduct this analysis, the 5 tapeworms from<br />
CRISCIONE AND FONT—VARIATION OF OOCHORISTICA JAVAENSIS 157<br />
each intensity level were treated as true replicates,<br />
when in fact they were pseudoreplicates.<br />
In addition, 10 relaxed tapeworms that were killed<br />
with hot water (90°C) were selected to provide measurements<br />
representative of O. javaensis recovered in<br />
a helminth survey of H. turcicus (C. D. Criscione, unpublished<br />
data). This representative data set included<br />
specimens from geckos with intensities ranging from<br />
1 to 64, and had at least 1 tapeworm from each of 5<br />
collection locations in southeastern Louisiana, U.S.A.<br />
[Bayou Segnette <strong>State</strong> Park in Westwego (29°53.18'N;<br />
90°09.80'W); Fairview-Riverside <strong>State</strong> Park in Madisonville<br />
(30°24.55'N; 90°08.41'W); a residential<br />
neighborhood in Metairie (30°00.76'N; 90°08.90'W);<br />
Southeastern Louisiana University in Hammond<br />
(30°30.67'N; 90°27.98'W); and LSU]. PCA with Varimax<br />
rotation was applied to this data set to examine<br />
for latent relationships among the same variables used<br />
in the fixation and intensity analyses.<br />
Specimens examined<br />
Museum specimens examined from the USNPC and<br />
the Canadian Museum of Nature (CMNPA), Ottawa,<br />
Ontario, Canada included the following: O. javaensis,<br />
2 paratypes (CMNPA nos. 1982-0693, 1982-0695);<br />
Oochoristica anolis Hardwood, 1932, 1 voucher<br />
(USNPC no. 75748) and the holotype (USNPC no.<br />
30898); Oochoristica bezyi Bursey and Goldberg,<br />
1992, 2 paratypes (USNPC no. 81874); Oochoristica<br />
bresslaui Fuhrmann, 1927, 1 voucher (USNPC no.<br />
89087); Oochoristica chinensis Jensen, Schmidt, and<br />
Kuntz, 1983, 2 paratypes (USNPC no. 0771<strong>68</strong>); Oochoristica<br />
islandensis Bursey and Goldberg, 1992, 1<br />
paratype (USNPC no. 82225); Oochoristica macallisteri<br />
Bursey and Goldberg, 1996, 1 voucher (USNPC<br />
no. 89267) and 2 paratypes (USNPC no. 86196); Oochoristica<br />
mccoyi Bursey and Goldberg, 1996, 2<br />
vouchers (USNPC nos. 85403, 85408) and 1 paratype<br />
(USNPC no. 86343); Oochoristica novaezealandae<br />
Schmidt and Allison, 1985, 5 paratypes (USNPC no.<br />
78407); Oochoristica osheroffi Meggitt, 1934, 1<br />
voucher (USNPC no. 80433); Oochoristica parvula<br />
(Stunkard, 1938), 3 vouchers (USNPC no. 84397);<br />
Oochoristica piankai Bursey, Goldberg, and Woolery,<br />
1996, 1 voucher (USNPC no. 88189) and 1 paratype<br />
(USNPC no. 84589); Oochoristica scelopori Voge and<br />
Fox, 1950, 2 vouchers (USNPC nos. 84234, 87529).<br />
We deposited voucher specimens of O. javaensis from<br />
H. turcicus in the USNPC (nos. 90344-90348).<br />
Results<br />
For the fixation analysis, PCA revealed 3 latent<br />
variables from the 8 measured (Table 1).<br />
Examination of the variable factor loadings revealed<br />
that factor 1 consisted of all the vertical<br />
measurements, while factors 2 and 3 were characterized<br />
by horizontal measures. Factors 1, 2,<br />
and 3 were renamed vertical, horizontal-1, and<br />
horizontal-2, respectively. All 3 factors showed<br />
significant worm-to-worm variation within each<br />
type of fixation (Fvcrtical = 9.20, FhorizontaM =<br />
Copyright © 2011, The Helminthological Society of Washington
158 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Table 1. Oochoristica javaensis: variable factor loadings, factor eigenvalues, and percent total variance<br />
accounted for by each factor from the Varimax rotated correlation matrix of the fixation data set.<br />
Testis length<br />
Ovary length<br />
Vitellaria length<br />
Proglottid length<br />
Ovary width<br />
Proglottid width<br />
Testis width<br />
Vitellaria width<br />
Eigenvalues<br />
Percent of total variance explained by the factor<br />
* Bold print shows loadings where variable loaded onto factor.<br />
42.20, Fhorizontal_2 = 5.31, df = 8, 20, P < 0.001);<br />
thus, the fixation main effect was tested with the<br />
mean-square values of the subgroups, tapeworms<br />
nested within fixation method. The vertical<br />
factor showed a significant effect between in<br />
situ-fixed and relaxed tapeworms (Fig. 1) (F]>8 =<br />
11.927, P = 0.009); however, neither horizontal<br />
factor was significant. Table 2 provides the raw<br />
measurements of the variables used in the analysis.<br />
PCA revealed that the 8 variables used in the<br />
intensity data set constituted only 1 factor (Table<br />
•3 0<br />
CJ<br />
1 -1<br />
-2<br />
In situ Relaxed<br />
Form of Fixation<br />
Figure 1. Plot of the mean factor scores and<br />
95% confidence intervals for the vertical factor of<br />
relaxed and in situ-fixed specimens of Oochoristica<br />
javaensis.<br />
Factor 1<br />
(vertical)<br />
0.917*<br />
0.875<br />
0.863<br />
0.805<br />
-0.203<br />
-0.236<br />
-0.092<br />
0.463<br />
3.320<br />
41.495<br />
Varimax rotated loading matrix<br />
Copyright © 2011, The Helminthological Society of Washington<br />
Factor 2<br />
(horizontal- 1)<br />
-0.112<br />
-0.176<br />
-0.237<br />
-0.497<br />
0.912<br />
0.869<br />
0.469<br />
0.177<br />
2.184<br />
27.296<br />
Factor 3<br />
(horiz.ontal-2)<br />
0.209<br />
0.209<br />
0.262<br />
0.023<br />
-0.082<br />
-0.096<br />
-0.835<br />
0.793<br />
1.498<br />
18.723<br />
3), thus showing that all 8 characters from relaxed<br />
specimens varied together. Even though<br />
there was significant worm-to-worm variation<br />
for this factor (F12 30 = 12.62, P < 0.001), there<br />
was still a significant intensity effect (F2 12 =<br />
13.42, P < 0.001) (Fig. 2). The a priori contrast<br />
between 15 and 28 was significant (F, 12 =<br />
10.58, P = 0.007), but 28 versus 64 was not.<br />
Figure 3A-C displays mature proglottid variation<br />
for tapeworms recovered from intensities of<br />
15, 28, and 64, and Table 4 gives the raw measurements<br />
of the variables. Measurements of 10<br />
O. javaensis tapeworms from Louisiana are given<br />
in Table 5. Based on results from the fixation<br />
experiments, only tapeworms exhibiting little to<br />
no wrinkling (i.e., contraction) were used to provide<br />
the representative measurements of O. javaensis<br />
collected in our survey. The fact that the<br />
8 variables in the representative data emerged as<br />
only 1 factor from the PCA (Table 3) again demonstrated<br />
that all 8 characters from relaxed specimens<br />
varied together.<br />
Discussion<br />
Variation resulting from different fixation<br />
techniques or conditions only confounds taxonomic<br />
problems in which the range of natural<br />
morphological variation is not known. Experimental<br />
data revealed 2 quantitative problems<br />
with using in situ-fixed tapeworms. The first was<br />
that a significant reduction in the vertical factor<br />
without a significant change in horizontal factors<br />
produced an accordion effect. High vertical factor<br />
scores were determined by large length measurements<br />
of the proglottid and its ovary, vitel-
CRISCIONE AND FONT—VARIATION OF OOCHORISTICA JAVAENSIS 159<br />
Table 2. Measurements of mature proglottids from specimens of Oochoristica javaensis fixed in a relaxed<br />
state and specimens fixed in situ; all measurements are in jxm.<br />
Proglottid width<br />
Proglottid length<br />
Ovary width<br />
Ovary length<br />
Vitellaria width<br />
Vitellaria length<br />
Testis width<br />
Testis length<br />
Sample size*<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
Relaxed<br />
n = 5t<br />
482-648 (589 ± 15.8)±<br />
356-640 (493 ± 25.8)<br />
246-35 1 (288 ± 8.06)<br />
152-238 (201 ± 6.63)<br />
125-195 (161 ± 5.11)<br />
90-137 (104 ± 3.83)<br />
31-43 (38.7 ± 0.83)<br />
27-47 (39.5 ± 1.23)<br />
* Sample size refers to the total number of each character that was measured.<br />
t Number of tapeworms used in measurements.<br />
:j: Range followed by mean ± 1 SE in parentheses.<br />
laria, and testes (Table 2); thus, relaxed tapeworms<br />
had a higher mean factor score (Fig. 1).<br />
Contraction from in situ fixation resulted in mature<br />
proglottids wider than long (Fig. 3D-F), but<br />
completely relaxed tapeworms from hot-fixed<br />
specimens yielded mature proglottids longer<br />
than wide (Fig. 3A-C).<br />
The second quantitative effect pertained to the<br />
correlative relationships among the mature proglottid<br />
characters and was revealed by the PCA<br />
itself. If only relaxed specimens are incorporated<br />
into the PCA, i.e., the intensity and representative<br />
data sets, all 8 characters vary together as<br />
Table 3. Oochoristica javaensis: variable factor<br />
loadings, factor eigenvalues, and percent total variance<br />
accounted for by each factor from the correlation<br />
matrix of the intensity data set and representative<br />
data set.<br />
Ovary width<br />
Vitellaria width<br />
Ovary length<br />
Proglottid width<br />
Vitellaria length<br />
Testis width<br />
Testis length<br />
Proglottid length<br />
Eigenvalues<br />
Percentage of total variance<br />
explained by the factor<br />
Loading matrices ;_<br />
for the O<br />
Intensity<br />
data set<br />
Factor 1<br />
0.924*<br />
0.921<br />
0.916<br />
0.893<br />
0.866<br />
0.841<br />
0.753<br />
0.751<br />
5.929<br />
74.116<br />
Representative<br />
Q<br />
data set "g<br />
Factor 1 W<br />
r~<br />
0.909 a<br />
0.854 ^<br />
0.814 ^<br />
0.851 ^<br />
0.784<br />
0.825<br />
0.749<br />
0.443<br />
4.996<br />
62.445<br />
Bold print shows loadings where variable loaded onto factor.<br />
In situ-fixed<br />
n = 5<br />
403-845 (635<br />
213-490 (312<br />
261-355 (310<br />
109-183 (144<br />
113-148 (130<br />
43-94 (70.5<br />
35-47 (43.5<br />
20-35 (27.1<br />
± 34.1)<br />
± 23.0)<br />
± 7.01)<br />
± 5.69)<br />
± 2.86)<br />
± 4.11)<br />
± 0.95)<br />
± 1.07)<br />
1 factor (Table 3); but when in situ-fixed tapeworms<br />
are incorporated into the PCA, i.e., the<br />
fixation data set, vertical measurements become<br />
independent of horizontal measurements (Table<br />
1). Contraction of the in situ-fixed tapeworms<br />
altered the correlative nature of the 8 variables<br />
and divided 1 factor into 3 factors.<br />
Contraction of helminth parasites resulting<br />
from improper fixation has been documented<br />
many times in the parasite literature (Bakke,<br />
1988); however, our study may be the first to<br />
quantify the effects of different forms of fixation<br />
and to analyze the data statistically. The empirical<br />
evidence provided in the current study not<br />
only supported the conclusions of Bakke (1988)<br />
1.5<br />
0.5<br />
-0.5<br />
-1.5<br />
15 28 64<br />
Intensity of Infection<br />
Figure 2. Plot of the mean factor scores and<br />
95% confidence intervals for tapeworms collected<br />
at intensities of 15, 28, and 64.<br />
Copyright © 2011, The Helminthological Society of Washington
160 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Figure 3. Mature proglottid variation of Oochoristica javaensis from Hemidactylus turcicus. Photomicrographs<br />
were taken with differential interference contrast. A-C. Natural variation of specimens from<br />
intensities of 15, 28, and 64, respectively; all were fixed in a relaxed state. D-F. Artificial variation showing<br />
contraction that resulted from in situ fixation. Bars = 200 u.m. CS = cirrus sac, GA = genital atrium, O<br />
= ovary, T = testis, and V = vitellaria.<br />
Table 4. Mature proglottid measurements of Oochoristica javaensis from Hemidactylus turcicus with intensities<br />
of 15, 28, and 64; all measurements are in |xm.<br />
Level of<br />
intensity<br />
Proglottid width<br />
Proglottid length<br />
Ovary width<br />
Ovary length<br />
Vitellaria width<br />
Vitellaria length<br />
Testis width<br />
Testis length<br />
Sample<br />
size*<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
15<br />
n<br />
15<br />
= 5t<br />
506-616 (560 ± 7.06):j:<br />
545-75 1 (624 ± 14.5)<br />
238-293 (2<strong>68</strong> ± 3.89)<br />
156-226 (189 ± 4.48)<br />
129-203 (161 ± 5.78)<br />
74-137 (105 ± 4.95)<br />
39-47 (42.7 ± 0.61)<br />
39-51 (43 d:<br />
0.87)<br />
28<br />
n = 5<br />
387-450 (409<br />
395-553 (470<br />
195-254 (225<br />
125-179 (160<br />
86-133 (117<br />
70-113 (90.3<br />
* Sample size refers to the total number of each character that was measured.<br />
t Number of tapeworms used in measurements.<br />
± Range followed by mean ± 1 SE in parentheses.<br />
± 5)<br />
± 14.7)<br />
± 4.85)<br />
± 3.83)<br />
± 3.37)<br />
± 3.3)<br />
35-43 (39 ± 0.78)<br />
31-47 (39 ± 1.03)<br />
Copyright © 2011, The Helminthological Society of Washington<br />
n<br />
269-545<br />
419-545<br />
121-281<br />
78-183<br />
59-117<br />
43-101<br />
27-43<br />
27-47<br />
64<br />
= 5<br />
(387<br />
(463<br />
(198<br />
(126<br />
± 31.7)<br />
± 8.7)<br />
± 14.2)<br />
± 7.9)<br />
(90.4 ± 5.19)<br />
(73.5 ± 4.91)<br />
(35 ± 1.29)<br />
(36.6 ± 1.4)
CRISCIONE AND FONT—VARIATION OF OOCHORISTICA JAVAENSIS 161<br />
Table 5. Measurements of Oochoristica javaensis from naturally infected Hemidactylus turcicus in southeastern<br />
Louisiana; measurements in jxin unless noted otherwise.<br />
Variable<br />
Total<br />
Proglottid number<br />
Neck<br />
Scolex<br />
Sucker<br />
Immature proglottid<br />
Genital pore position*<br />
Mature proglottid<br />
Cirrus sac<br />
Ovary<br />
Vitellaria<br />
Testis<br />
Testes number<br />
Gravid proglottid<br />
Oncosphere<br />
Hook<br />
Li (mm)<br />
W<br />
L (mm)<br />
W<br />
L<br />
W<br />
L<br />
W<br />
L<br />
W<br />
L<br />
W<br />
L<br />
W<br />
L<br />
W<br />
L<br />
W<br />
L<br />
W<br />
L (mm)<br />
W<br />
L<br />
L<br />
Sample size*<br />
10<br />
10<br />
10<br />
10<br />
10<br />
10<br />
10<br />
10<br />
30<br />
30<br />
30<br />
30<br />
30<br />
30<br />
30<br />
30<br />
30<br />
30<br />
30<br />
30<br />
30<br />
30<br />
30<br />
30<br />
30<br />
30<br />
30<br />
n = 10t<br />
22.2-105 (53.4 ± 7.4)§,||<br />
86-164 (131 ± 7.8)||<br />
158-237 (205 ± 8.1)||<br />
1.12-1.58 (1.38 ± 0.05)||<br />
148-246 (195 ± 9.1)||<br />
98-183 (140 ± 8.9)<br />
51-90 (74.1 ± 3.9)||<br />
62-117 (89 ± 5.3)||<br />
261-506 (408 ± 14.5)<br />
237-371 (297 ± 7.6)<br />
0.24-0.33 (0.28 ± 0.004)||<br />
277-648 (490 ± 19.5)||<br />
395-751 (509 ± 17.8)||<br />
43-55 (47.7 ± 0.58)<br />
86-144 (116 ± 2.8)||<br />
133-390 (2<strong>68</strong> ± 11.8)||<br />
78-316 (184 ± 0.6)||<br />
62-269 (144 ± 8.9)||<br />
43-221 (106 ± 6.6)||<br />
27-51 (40.2 ± 0.98)||<br />
31-47 (40.9 ± 0.9)<br />
17-46 (26.6 ± 1.4)||<br />
158-650 (492 ± 29.4)||<br />
0.85-1.99 (1.26 ± 0.05)||<br />
20-34 (25.3 ± 0.69)||<br />
18-28 (23.2 ± 0.52)||<br />
8-12 (11.5 ± 0.18)||<br />
* Sample size refers to the total number of each character that was measured.<br />
t Number of tapeworms measured.<br />
± L = length, W = width.<br />
§ Range followed by mean ± 1 SE in parentheses.<br />
|| Indicates that the range of the character extends values reported in the original description (Kennedy et al., 1982).<br />
# Genital pore position was calculated as a ratio of the position along the length of the mature proglottid from the anterior<br />
end (length to the center of the genital pore + length of proglottid).<br />
but also quantitatively demonstrated the misleading<br />
representation of morphological characters<br />
used in the taxonomy of Oochoristica resulting<br />
from in situ fixation. We believe that our<br />
more rigorous analysis is important because, despite<br />
the elegant studies of Bakke (1988) and<br />
previous workers, the practice of describing improperly<br />
fixed specimens is still widespread<br />
among parasite taxonomists.<br />
In addition to the quantitative changes resulting<br />
from in situ fixation, 2 qualitative effects further<br />
demonstrated the inappropriateness of using<br />
in situ-fixed specimens in species descriptions.<br />
Distortion of the scolex (Fig. 4A, B) and proglottids<br />
(Fig. 4C, D) prevented accurate measurements<br />
of these characters. This is not to say<br />
that every scolex and mature proglottid fixed in<br />
situ will be rendered useless for species identi-<br />
fication, but the number of appropriate characters<br />
available for analysis will be greatly reduced.<br />
As seen in Figure 4A-D, one would have<br />
difficulty in finding a true scolex width, and contortion<br />
in the strobila would limit the number of<br />
proglottids suitable for examination.<br />
Species descriptions based on contracted or<br />
disfigured specimens will misrepresent the true<br />
natural variation by decreasing the means of vertical<br />
characters and artificially inflating the dispersion<br />
of measurements if used in conjunction<br />
with relaxed specimens. Such may be the case<br />
with several paratype (O. bezyi, O. islandensis,<br />
O. macallisteri, O. mccoyi, O. piankai} and<br />
voucher (O. mccoyi, O. par~vula, O. piankai, O.<br />
scelopori) specimens examined in our study.<br />
These specimens may represent true species, but<br />
their reported natural variation is more than like-<br />
Copyright © 2011, The Helminthological Society of Washington
162 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Figure 4. Comparisons of relaxed and in situ-fixed specimens of Oochoristicajavaensis. A-D. Distorting<br />
effects of in situ fixation on scolices and proglottids. E-H. Relaxed specimens. Bars = 200 (xm for A, B,<br />
E, and F (differential interference contrast). Bars = 500 (Jim for C, D, G, and H (brightfield).<br />
ly masked within the artificial variation induced<br />
by in situ fixation. Characters that do not reflect<br />
their natural variation should not be used to describe<br />
species. Oochoristica spp. recovered from<br />
fixed museum hosts may provide historical<br />
abundance data, but identification of these specimens<br />
should be made with extreme caution, and<br />
ideally, in conjunction with specimens fixed appropriately.<br />
Intensity effects were examined because a<br />
crowding effect has been documented as a cause<br />
of variation in the size of tapeworms (Read,<br />
1951), and because of the occurrence of different<br />
intensities in naturally infected H. turcicus.<br />
PCA for the intensity data set produced 1 factor<br />
in which all 8 variables loaded high (Table 3);<br />
therefore, individual proglottids with large measurement<br />
values received high factor scores. Although<br />
not quantified, there was no apparent<br />
crowding effect observed for natural infections<br />
with intensities between 1 and 15. Tapeworms<br />
from an intensity of 15 had significantly greater<br />
factor scores than specimens from 28 and 64<br />
(Fig. 2), thus indicating that crowding reduces<br />
the size of the respective morphological characters<br />
(Table 4) in O. javaensis. Brooks and<br />
Copyright © 2011, The Helminthological Society of Washington<br />
Mayes (1976) reported similar crowding effects<br />
for O. bivitellobata recovered from the prairie<br />
racerunner, Cnemidophorus sexlineatus Lowe,<br />
1966. Their results and ours, however, should be<br />
considered only preliminary for 2 reasons. First,<br />
data were obtained from natural infections; thus<br />
other factors that induce variation were not controlled.<br />
Second, the intensity levels were not<br />
replicated. Ideally, one would wish to sample<br />
tapeworms from multiple hosts harboring all<br />
possible intensity levels. Both reports, however,<br />
suggest that morphological characters for species<br />
of Oochoristica can be variable and may be<br />
subject to intensity levels.<br />
Measurements of the 10 0. javaensis specimens<br />
given in Table 5 extend the ranges of several<br />
characters provided in the original description<br />
of O. javaensis (Kennedy et al., 1982).<br />
These measurements were based on specimens<br />
with little or no contraction and are provided to<br />
give a representation of O. javaensis collected<br />
from H. turcicus in southeastern Louisiana.<br />
Means of several characters (Table 5) do not<br />
match those provided by Kennedy et al. (1982);<br />
however, based on the lack of host specificity<br />
displayed in laboratory experiments (Criscione
and Font, <strong>2001</strong>), the indication of plasticity in<br />
morphological characters (Table 4), and the examination<br />
of O. javaensis paratypes, it was determined<br />
that the specimens from H. turcicus in<br />
southeastern Louisiana were O. javaensis.<br />
In summary, statistical analyses demonstrated<br />
that measurements of in situ-fixed tapeworms,<br />
i.e., specimens recovered from preserved hosts,<br />
distorted the true natural variation of O. javaensis.<br />
The intraspecific variation of several species<br />
of Oochoristica may be misrepresented because<br />
they were described from highly contracted, in<br />
situ-fixed specimens. Additionally, morphological<br />
characters used in the taxonomy of Oochoristica<br />
have not been examined for their stability<br />
when exposed to different environmental<br />
or host-induced conditions. Our analyses indicated<br />
that proglottid morphology was highly<br />
variable and that this plasticity may have resulted<br />
from crowding.<br />
Acknowledgments<br />
We extend our thanks to Dr. Murray Kennedy<br />
and Dr. Bruce Conn for loaning us specimens of<br />
Oochoristica from their private collections, and to<br />
Judith Price at the CMNPA and Pat Pilitt, Dr. Eric<br />
Hoberg, and Dr. Ralph Lichtenfels at the USNPC<br />
for their assistance and loan of specimens.<br />
CRISCIONE AND FONT—VARIATION OF OOCHORISTICA JAVAENSIS 163<br />
Notice of Dues Increase<br />
Literature Cited<br />
Bakke, T. A. 1988. Morphology of adult Phyllodistomum<br />
umblae (Fabricius) (Platyhelminthes, Gorgoderidae):<br />
the effect of preparation, killing and<br />
fixation procedures. Zoologica Scripta 17:1-13.<br />
Brooks, D. R., and M. A. Mayes. 1976. Morphological<br />
variation in natural infections of Oochoristica<br />
bivitellobata Loewen, 1940 (Cestoidea: Anoplocephalidae).<br />
Transactions of the Nebraska Academy<br />
of Sciences 3:20-21.<br />
Criscione, C. D., and W. F. Font. <strong>2001</strong>. Development<br />
and specificity of Oochoristica javaensis (Eucestoda:<br />
Cyclophyllidea: Anoplocephalidae: Linstowiinae).<br />
<strong>Comparative</strong> <strong>Parasitology</strong> <strong>68</strong>:149-155.<br />
Hair, J. F., Jr., R. E. Anderson, R. L. Tatham, and<br />
W. C. Black. 1999. Multivariate Data Analysis,<br />
5th ed. Prentice-Hall, Upper Saddle River, New<br />
Jersey, U.S.A. 730 pp.<br />
Haley, A. J. 1962. Role of host relationships in the<br />
systematics of helminth parasites. Journal of <strong>Parasitology</strong><br />
48:671-678.<br />
Kennedy, M. J., L. M. Killick, and M. Beverley-<br />
Burton. 1982. Oochoristica javaensis n. sp. (Eucestoda:<br />
Linstowiidae) from Gehyra mutilata and<br />
other gekkonid lizards (Lacertilia: Gekkonidae)<br />
from Java, Indonesia. Canadian Journal of Zoology<br />
60:2459-2463.<br />
Read, C. P. 1951. The "crowding effect" in tapeworm<br />
infections. Journal of <strong>Parasitology</strong> 37:174-178.<br />
Stunkard, H. W. 1957. Intraspecific variation in parasitic<br />
flatworms. Systematic Zoology 6:7-18.<br />
At the January <strong>2001</strong> meeting of the Helminthological Society of Washington, the Society voted to<br />
increase the membership dues to US$30.00 for individual U.S. members and to US$33.00 for individual<br />
foreign members. Institution subscription rates will increase as follows: US$55.00 (U.S.A.), US$57.00<br />
(Canada and Mexico), US$60.00 (all other countries). This is the first dues increase in six years and is<br />
necessitated by increased management costs to the Society. The increases will take effect in January, 2002.<br />
Copyright © 2011, The Helminthological Society of Washington
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 164-172<br />
Seasonal Occurrence and Community Structure of Helminth<br />
Parasites in Green Frogs, Rana clamitans melanota, from<br />
Southeastern Wisconsin, U.S.A.<br />
MATTHEW G. BOLEK' AND JAMES R. COGGINS<br />
Department of Biological Sciences, University of Wisconsin-Milwaukee, Milwaukee, Wisconsin 53201,<br />
U.S.A. (e-mail: coggins@csd.uwm.edu)<br />
ABSTRACT: From April to October 1996, 75 green frogs, Rana clamitans melanota, were collected from Waukesha<br />
County, Wisconsin, U.S.A. and examined for helminth parasites. Seventy-one (94%) of 75 green frogs<br />
were infected with 1 or more helminth species. The component community consisted of 12 species: 5 trematodes,<br />
2 cestodes, and 5 nematodes. Approximately 2,790 (72%) trematodes, 447 (11.5%) cestodes, and 636 (16.5%)<br />
nematodes were found. A significant correlation existed between wet weight and helminth species richness.<br />
Helminth populations and communities were seasonally variable and/or did not show significant differences<br />
during the year. Haematoloechus varioplexus showed seasonal variation in size during the year that was related<br />
to recruitment period. The helminth fauna of green frogs was depauperate and dominated by indirect-life-cycle<br />
parasites. Host diet and aquatic habitat were important in the transmission dynamics of these species. Host size,<br />
sex, and time of collection were also important factors in structuring helminth communities of green frogs and<br />
may mask any simple explanations.<br />
KEY WORDS: Rana clamitans, Haematoloechus varioplexus, Halipegus eccentricus, Glyptheltnins quieta, Cosmocercoides<br />
sp., Oswaldocruzia pipiens, Waltonella sp., Mesocestoides sp., metacercariae, Trematoda, Nematoda,<br />
Cestoda, Amphibia, seasonal study, Wisconsin, U.S.A.<br />
Studies by Kennedy et al. (1986) on freshwater<br />
fish, birds, and a mammal have developed<br />
predictions in determining helminth community<br />
structure, particularly that ectotherm and endotherm<br />
helminth communities are fundamentally<br />
different: the former are species poor and noninteractive,<br />
while the latter are diverse and interactive.<br />
A review by Aho (1990) indicated that<br />
helminth communities of amphibians are highly<br />
variable, depauperate, and noninteractive in<br />
structure, but there is a need to examine and<br />
reexamine more species from different locations.<br />
To date there are few studies that utilized helminth<br />
community measures in amphibian hosts<br />
(Goater et al., 1987; Aho, 1990; Muzzall, 1991a,<br />
b; Goldberg et al., 1995; Barton, 1996; Yoder<br />
and Coggins, 1996; McAlpine, 1997; Bolek and<br />
Coggins, 2000). A number of helminth surveys<br />
of green frogs, Rana clamitans melanota Rafinesque,<br />
1820, have been published (Campbell,<br />
19<strong>68</strong>; Williams and Taft, 1980; Coggins and Sajdak,<br />
1982; McAlpine and Burt, 1998), but there<br />
are few studies on the helminth infracommunity<br />
and component community structure of this spe-<br />
1 Corresponding author. Current address: School of<br />
Biological Sciences, University of Nebraska-Lincoln,<br />
Lincoln, Nebraska <strong>68</strong>588, U.S.A. (e-mail: mbolek@<br />
unlserve.unl.edu).<br />
164<br />
Copyright © 2011, The Helminthological Society of Washington<br />
cies (Muzzall, 199la; McAlpine, 1997), and<br />
none that incorporated a seasonal component.<br />
Green frogs are large, semiaquatic frogs inhabiting<br />
freshwater ponds, lakes, swamps, and<br />
slow-moving streams in North America. They<br />
spend most of their time around the water's<br />
edge. They occur from Newfoundland to western<br />
Ontario, and south to eastern Oklahoma,<br />
southern Illinois, northern Georgia, and eastern<br />
North Carolina. In Wisconsin, these frogs overwinter<br />
buried in the mud and are active from<br />
early April through October (Vogt, 1981). Green<br />
frogs are largely sit-and-wait gape-limited predators,<br />
feeding on any accessible prey of appropriate<br />
size, including aerial, aquatic, and terrestrial<br />
invertebrates, primarily insects (Scale,<br />
1987; Werner et al., 1995). Here we report on<br />
the seasonal helminth community structure of<br />
green frogs from southeastern Wisconsin. Specifically,<br />
we were interested in how host habitat,<br />
age and/or size, diet, sex, and seasonality were<br />
important in determining helminth populations<br />
and communities of green frogs.<br />
Materials and Methods<br />
A total of 75 green frogs, R. clamitans melanota,<br />
was collected from April to October 1996 at a small<br />
spring-fed permanent pond located at the Carroll <strong>College</strong><br />
field station in Waukesha County, Wisconsin,
U.S.A. (42°59'N; 88°21'W). Ten to 15 frogs were collected<br />
monthly around the periphery of the pond by a<br />
dip-net. Animals were placed in plastic containers,<br />
transported to the laboratory, stored at 4°C, and euthanized<br />
in MS 222 (ethyl m-aminoben/.oate methane sulfonic<br />
acid) within 72 hr of capture. Snout-vent length<br />
(SVL) and wet weight (WW) were recorded for each<br />
individual. Frogs were individually toe-clipped and<br />
frozen. At necropsy, the digestive tracts, limbs and<br />
body wall musculature, and internal organs were examined<br />
for helminth parasites. Each organ was placed<br />
individually in a petri dish and examined under a stereomicroscope.<br />
The body cavities were rinsed with distilled<br />
water into a petri dish and the contents examined.<br />
All individuals were sexed by gonad inspection during<br />
necropsy. Worms were removed and fixed in alcohol—<br />
formaldehyde-acetic acid or formalin. Trematodes and<br />
cestodes were stained with acetocarmine, dehydrated<br />
in a graded ethanol series, cleared in xylene, and<br />
mounted in Canada balsam. Nematodes were dehydrated<br />
to 70% ethanol, cleared in glycerol, and identified<br />
as temporary mounts. Prevalence, mean intensity,<br />
and abundance are according to Bush et al. (1997).<br />
Mean intensity was not calculated for the unidentified<br />
kidney metacercariae because they could not be counted<br />
accurately, and overall abundance was reported as<br />
an estimate of encysted metacercariae counted on the<br />
surface of the kidneys. Mean helminth species richness<br />
is the sum of helminth species per individual frog, including<br />
noninfected individuals, divided by the total<br />
sample size. All values are reported as the mean ± 1<br />
standard deviation. Undigested stomach contents were<br />
identified to class or order following Borror et al.<br />
(1989). Stomach contents are reported as a percent =<br />
the number of prey items in a given class or order,<br />
divided by the total number of prey items recovered<br />
X 100. Voucher specimens have been deposited in the<br />
H. W. Manter Helminth Collection, University of Nebraska,<br />
Lincoln, Nebraska, U.S.A. (accession numbers<br />
HWML 15354, Haematoloechus varioplexus Stafford,<br />
1902; 15355, Glypthelmins quieta Stafford, 1900;<br />
15356, kidney metacercariae; 15357, Mesocestoides<br />
sp.; 15358, diplostomid metacercariae; 15359, Halipegus<br />
eccentricus Thomas, 1939; 15360, unidentified<br />
adult tapeworm; 15361, Cosmocercoides sp.; 15362,<br />
unidentified larval nematode; 15363, unidentified species<br />
of Waltonella Schacher, 1974; 15364, Oswaldocruzia<br />
pipiens Walton, 1929).<br />
The chi-square test for independence was calculated<br />
to compare differences in prevalence among host sex.<br />
Yates' adjustment for continuity was used when sample<br />
sizes were low. A single-factor, independent-measures<br />
ANOVA and Scheffe's posthoc test were used to<br />
compare among seasonal differences in mean intensity<br />
and mean helminth species richness. When variances<br />
were heteroscedastic, the Kruskal-Wallis test and the<br />
Kolmogorov—Smirnov two-sample test were used. Student's<br />
?-test was used to compare differences in mean<br />
intensity and mean helminth species richness between<br />
sex of hosts. Approximate Mests were calculated when<br />
variances were heteroscedastic (Sokal and Rohlf,<br />
1981). Pearson's correlation was used to determine relationships<br />
among host SVL and WW and abundance<br />
of helminth parasites, excluding larval platyhelminths.<br />
BOLEK AND COGGINS—HELMINTH COMMUNITIES IN GREEN FROGS 165<br />
Pearson's correlation was calculated for host SVL and<br />
WW and helminth species richness per individual frog.<br />
Because WW gave a stronger correlation than SVL in<br />
each case, it is the only parameter reported. Because<br />
of low sample size during certain collection periods,<br />
data were pooled on a bimonthly basis to form samples<br />
of 15—20 frogs per season. Larval helminths were not<br />
included in the seasonal analysis, because they can<br />
also accumulate throughout the amphibian's life and<br />
thus mask monthly recruitment dynamics in adult<br />
frogs.<br />
Results<br />
A total of 75 adult green frogs, 43 males and<br />
32 females, was collected during April through<br />
October 1996. No significant difference existed<br />
in the number of male and female frogs collected<br />
throughout the year (x2 = 7.01, P > 0.05).<br />
The overall means of SVL and WW of green<br />
frogs were <strong>68</strong>.8 ± 10.8 mm (range 39.8-89.4<br />
mm) and 35.8 ± 15.5 g (5.4-75.6 g), respectively.<br />
There was no significant difference in<br />
mean SVL (t = 0.10, P > 0.05) or mean WW<br />
(t = 0.24, P > 0.05) in male and female frogs.<br />
Stomach content analyses of green frogs revealed<br />
a broad range of aerial, terrestrial, and<br />
aquatic invertebrates. Sixteen different groups of<br />
invertebrates were recovered from stomach contents<br />
of green frogs, with coleopterans, gastropods,<br />
and diplopodans making up the largest<br />
percentage.<br />
Seventy-one (94%) of 75 R. clamitans melanota<br />
were infected with helminth parasites. The<br />
component community consisted of 12 species<br />
(5 trematodes, 2 cestodes, and 5 nematodes). Of<br />
these, 8 have indirect life cycles, 1 has a direct<br />
life cycle, and the life cycles of 3 are unknown.<br />
Overall mean helminth abundance, excluding<br />
larval platyhelminths, was 16.5 ± 38 with most<br />
frog infracommunities having 10 or fewer<br />
worms. In terms of abundance, digeneans dominated<br />
adult helminth communities (61.5% of total<br />
adult helminths). Prevalence ranged from<br />
80% for kidney metacercariae to 1.3% for an<br />
unidentified adult cestode, a filarid nematode of<br />
the genus Waltonella, and an unidentified encysted<br />
nematode. Values for overall prevalence,<br />
mean intensity, mean abundance, and total number<br />
of helminths recovered are summarized in<br />
Table 1.<br />
Statistically significant differences in prevalence<br />
or mean intensity existed between male<br />
and female frogs for H. varioplexus, H. eccentricus,<br />
kidney metacercariae, and unidentified<br />
species of Mesocestoides Valunt, 1863 and Cos-<br />
Copyright © 2011, The Helminthological Society of Washington
166 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Table 1. Prevalence, mean intensity (MI), mean abundance (MA), and total helminths found in 75 specimens<br />
of Rana clamitans melanota in Wisconsin.<br />
Trematoda<br />
Species<br />
Haematoloechus varioplexus<br />
Halipegus eccentricus<br />
Glypthelmins quieta<br />
Unidentified metacercariae:i:<br />
Diplostomid metacercariae*<br />
Cestoda<br />
Unidentified adult cestode<br />
Mesocestoides sp.*<br />
Nematoda<br />
Oswaldocruzia pipiens<br />
Cosmoccrcoides sp.<br />
Waltonclla sp.t<br />
Larval nematode<br />
Encysted nematode<br />
* Underestimate.<br />
t New host record.<br />
± Not counted.<br />
Prevalence: MI ± 1 SD<br />
Number of<br />
worms<br />
number (%) (range) MA ± 1 SD recovered<br />
33 (44)<br />
17 (22.6)<br />
11 (14.6)<br />
60 (80)<br />
12 (16.0)<br />
1 (1.3)<br />
14 (18.6)<br />
21 (28)<br />
19 (25)<br />
1 (1.3)<br />
9 (12)<br />
1 (1.3)<br />
5.3 ± 7<br />
d-30)<br />
1.9 ± 1.7<br />
(1-8)<br />
35.5 ± 34.7<br />
(1-110)<br />
NC:i:<br />
35 ± 41<br />
(1-100)<br />
1<br />
31.9 ± 22<br />
(12-86)<br />
14.7 ± 23<br />
(1-90)<br />
3.3 ± 4.3<br />
(1-19)<br />
4<br />
25.9 ± 61<br />
(1-200)<br />
1<br />
mocercoides Harwood, 1930 (Table 2). Both H.<br />
varioplexus, a lung trematode, and H. eccentricus,<br />
a trematode of the eustachian tubes, had significantly<br />
higher mean intensities in male frogs,<br />
while the kidney metacercariae occurred at a<br />
higher prevalence in males. Female frogs had<br />
significantly higher mean intensities of Mesocestoides<br />
sp. and Cosmocercoides sp.<br />
Mean helminth species richness was 2.<strong>68</strong> ±<br />
1.29 species per frog. Infections with multiple<br />
species were common, with 0, 1,2, 3, 4, and 5<br />
species occurring in 4, 8, 23, 20, 13, and 7 frogs,<br />
respectively. No statistically significant differences<br />
in mean helminth species richness were<br />
found between male (2.76 ± 1.01) and female<br />
frogs (2.56 ± 1.52, t = 0.66, P > 0.05). A nonsignificant<br />
positive correlation was found between<br />
overall helminth abundance, excluding<br />
larval platyhelminths, and WW (/• = 0.04, P ><br />
0.05). Nonsignificant relationships were also observed<br />
for most helminth species: unidentified<br />
adult cestode (r = -0.01, P > 0.05), H. vario-<br />
2.3 ± 5.4<br />
0.4 ± 1.1<br />
5.2 ± 18<br />
NC<br />
5.7 ± 20.7<br />
0.01 ± 0.1<br />
5.9 ± 15.3<br />
4.1 ± 13.7<br />
0.8 ± 2.6<br />
0.05 ± 0.5<br />
3.5 ± 23<br />
0.01<br />
± 0.1<br />
176<br />
33<br />
391<br />
> 1,770<br />
420<br />
1<br />
446<br />
310<br />
62<br />
4<br />
259<br />
Copyright © 2011, The Helminthological Society of Washington<br />
1<br />
Lungs<br />
Location<br />
Eustachian tubes<br />
Small intestine<br />
Kidneys, body cavity<br />
Leg muscles<br />
Small intestine<br />
Leg muscles, lungs<br />
Small intestine, stomach<br />
Large intestine, small intestine<br />
Body cavity<br />
Large intestine<br />
Small intestine<br />
plexus (r = 0.10, P > 0.05), H. eccentricus (r<br />
= 0.19, P > 0.05), G. quieta (r = -0.02, P ><br />
0.05), O. pipiens (r = -0.12, P > 0.05), unidentified<br />
larval nematode (r = -0.04, P ><br />
0.05), Waltonella sp. (r = 0.20, P > 0.05), and<br />
encysted nematodes (r = -0.19, P > 0.05). The<br />
nematode Cosmocercoides sp. had a significant<br />
positive correlation with WW (/• = 0.31, P <<br />
0.01). A significant positive Pearson's correlation<br />
also existed between species richness and<br />
WW (r = 0.31, P < 0.01). However, correlations<br />
between frog WW and species richness<br />
were not significant in May—June (/• = 0.31, P<br />
> 0.05), July-August (r = 0.01, P > 0.05), and<br />
September-October (/- = 0.29, P > 0.05) but<br />
were significant for the April collection (r =<br />
0.60, P < 0.02).<br />
The trematodes H. varioplexus and H. eccentricus<br />
occurred throughout the year, with highest<br />
prevalences observed during the fall (September-October)<br />
collection, 65% and 30%, respectively.<br />
The intestinal trematode, G. quieta, was
Table 2. Prevalence (Pr) and<br />
Rana clamitans melanota.<br />
Trematoda<br />
Species<br />
Haematoloechus varioplexus<br />
Halipegns eccentricus<br />
Glypthelmins quieta<br />
Unidentified metacercariae*<br />
Diplostomid metacercariae*<br />
Cestoda<br />
Unidentified adult cestode<br />
Mesocestoides sp.*<br />
Nematoda<br />
Oswaldocruzia pipiens<br />
Cosmocercoides sp.<br />
Waltonella sp.<br />
Larval nematode<br />
Encysted nematode<br />
* Underestimate.<br />
± Not counted.<br />
BOLEK AND COGGINS—HELMINTH COMMUNITIES IN GREEN FROGS 167<br />
mean intensity (MI) of helminth parasites in male and female green frogs,<br />
Measure of<br />
parasitism<br />
Pr<br />
MI ±<br />
Pr<br />
MI ±<br />
PI-<br />
MI ±<br />
Pr<br />
MI ±<br />
Pr<br />
MI ±<br />
Pr<br />
MI ±<br />
Pr<br />
MI ±<br />
Pr<br />
MI ±<br />
Pr<br />
MI ±<br />
Pr<br />
MI ±<br />
Pr<br />
MI ±<br />
PI-<br />
MI ±<br />
1 SD<br />
1 SD<br />
1 SD<br />
1 SD<br />
1 SD<br />
1 SD<br />
1 SD<br />
1 SD<br />
1 SD<br />
1 SD<br />
1 SD<br />
1 SD<br />
first observed during midspring (May-June)<br />
with a prevalence of 5%. Prevalence for this<br />
species reached its maximum (30%) during summer<br />
(July-August) and decreased during the fall<br />
collection (20%). Seasonal mean intensity of<br />
adult platyhelminths followed similar patterns as<br />
prevalence, but no significant differences existed<br />
for any of the adult platyhelminths recovered, H.<br />
eccentricus (adjusted H = 2.02, P > 0.05), H.<br />
varioplexus (F = 0.34, P > 0.05), or G. quieta<br />
(t = 0.43, P > 0.05).<br />
Although prevalence and intensity of H. varioplexus<br />
did not vary significantly throughout<br />
the collection period, mean length of worms did<br />
(Fig. 1). Greatest mean length of worms (4.1<br />
mm) was recorded in early spring (April), when<br />
all individuals were gravid adults, and reached<br />
a minimum during midspring (1.84 mm), when<br />
immature individuals were common. Statistically<br />
significant differences in mean length were<br />
observed for April and May—June collections,<br />
Males<br />
N = 43<br />
46.5 (20/43)<br />
7 ± 8.6<br />
23.3 (10/43)<br />
2.3 ± 2.1<br />
1 1 .6 (5/43)<br />
21.8 ± 27.9<br />
90.7 (39/43)<br />
NC:!:<br />
18.6 (8/43)<br />
45.3 ± 46.1<br />
2.3 (1/43)<br />
1<br />
23.3 (10/43)<br />
24.9 ± 16.2<br />
27.9 (12/43)<br />
16.6 ± 25.8<br />
20.9 (9/43)<br />
2+1.7<br />
0 (0/43)<br />
0<br />
9.3 (4/43)<br />
11.5 ± 11.2<br />
0 (0/43)<br />
0<br />
Females<br />
N = 32<br />
40.6 ( 1 3/32)<br />
2.8 ± 2.2<br />
21.9 (7/32)<br />
1.4 ± 0.5<br />
18.8 (6/32)<br />
47 ± 38.1<br />
65.6 (21/32)<br />
NC<br />
12.5 (4/32)<br />
14.5 ± 23.7<br />
0 (0/32)<br />
0<br />
12.5 (4/32)<br />
49.3 ± 24.8<br />
28.1 (9/32)<br />
12.2 ± 19.6<br />
31.3 (10/32)<br />
4.4 ± 5.6<br />
3.1 (1/32)<br />
4<br />
18.8 (6/32)<br />
35.5 ± 80.6<br />
3.1 (1/32)<br />
1<br />
Statistic<br />
X2 = 0.24<br />
f',= 3.07<br />
X2 = 0.02<br />
f',= 2.71<br />
X2 = 0.46<br />
t = 1 .35<br />
X2 = 5.72<br />
X2odj = 0.16<br />
t = 1.23<br />
X2ailJ = 0.02<br />
X2adj = 0.78<br />
t = 6.69<br />
x2 = o.oo<br />
/ = 0.44<br />
X2 = LOO<br />
r',= 3.81<br />
X2ailj = 0.02<br />
X2:,Ji = 1-67<br />
f',= 1.67<br />
X2a(lj = 0.02<br />
P<br />
>0.05<br />
0.05<br />
0.05<br />
>0.05<br />
0.05<br />
>0.05<br />
>().05<br />
>0.05<br />
0.05<br />
>0.05<br />
>0.05<br />
0.05<br />
>0.05<br />
>0.05<br />
>0.05<br />
April and July-August collections, May-June<br />
and September—October collections, and July-<br />
August and September-October collections (F =<br />
11.8, P < 0.05, single-factor, independent-measure<br />
ANOVA; P < 0.05 for all pair-wise comparisons,<br />
Scheffe's test).<br />
The nematodes Waltonella sp. and an unidentified<br />
encysted larva were recovered infrequently<br />
as single infections during midspring and fall<br />
collections, respectively. Prevalence and intensity<br />
values for O. pipiens, Cosmocercoides sp.,<br />
and unidentified larval nematodes were low and/<br />
or erratic over the 7-mo period. The nematodes<br />
O. pipiens, Cosmocercoides sp., and unidentified<br />
larval nematodes were first observed during<br />
midspring and persisted until fall, with prevalence<br />
being highest in summer for O. pipiens<br />
(62%) and midspring and summer for Cosmocercoides<br />
sp. (40%) and larval nematodes<br />
(20%). However, only O. pipiens exhibited statistically<br />
significant differences (adjusted H =<br />
Copyright © 2011, The Helminthological Society of Washington
1<strong>68</strong> COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
7 -r<br />
N = 20 N = 27 N = 28 N = 79<br />
Apr May-Jun Jul- Aug Sep-Oct<br />
Date<br />
Figure 1. Mean lengths of Haematoloechus varioplexus<br />
from Rana clamitans melanota. N = number<br />
of worms measured from all frogs recovered in<br />
each sampling period.<br />
9.45, P < 0.05) in mean intensity. The two-sample<br />
Kolmogorov-Smirnov test revealed significant<br />
differences in mean intensities during the<br />
May-June (27 ± 30) and July-August (9.8 ±<br />
17) collections.<br />
Mean helminth species richness fluctuated<br />
seasonally (Fig. 2) and was lowest (1.53) during<br />
early spring and highest (3.1) during the summer<br />
collections. Statistically significant differences<br />
in mean helminth species richness were observed<br />
for April and May—June collections,<br />
April and July—August collections, and April<br />
and September-October collections (F = 6.01,<br />
P < 0.05, single-factor, independent-measure<br />
ANOVA; P < 0.05 for all pairwise comparisons,<br />
Scheffe's test). No statistically significant differences<br />
in frog WW were observed during the<br />
year (F = 0.37, P > 0.05).<br />
Discussion<br />
Wisconsin green frogs had high overall helminth<br />
prevalence, with parasite infracommunities<br />
being dominated by indirect life-cycle parasites.<br />
Of the identified parasites, only 1 direct<br />
life-cycle nematode, O. pipiens, was present,<br />
with most helminth species displaying a prevalence<br />
below 50% and/or low mean intensities<br />
below 30.<br />
Most green frogs contained identifiable stom-<br />
5-r<br />
Copyright © 2011, The Helminthological Society of Washington<br />
= 15 N = 20 N = 20 N = 20<br />
May-Jim Jul-Aug Sep-Oct<br />
Date<br />
Figure 2. Mean helminth species richness of<br />
Rana clamitans melanota during April, May-June,<br />
July-August, and September-October 1996. N =<br />
number of frogs collected on each date.<br />
ach contents containing predominantly beetles,<br />
gastropods, and diplopods. In total, 16 different<br />
groups of terrestrial, aerial, and aquatic invertebrates<br />
comprised their stomach contents.<br />
These results appear similar to those of other<br />
investigators (Hamilton, 1948; Stewart and Sandison,<br />
1972; McAlpine and Dilworth, 1989;<br />
Werner et al., 1995). Hamilton (1948) found that<br />
the principal foods of green frogs collected in<br />
New York, U.S.A. consisted of beetles, flies, and<br />
grasshoppers, with a total of 15 different items<br />
recovered from adult frogs and 20 different prey<br />
items recovered from various sized individuals.<br />
The most common helminth recovered was an<br />
unidentified kidney metacercaria. This larval<br />
trematode had an overall prevalence of 80% and<br />
mean intensity of over 30 worms per frog. Four<br />
other digeneans were recovered from green<br />
frogs: a diplostomid metacercaria encysted in<br />
the musculature, and 3 adult trematodes: H. varioplexus,<br />
H. eccentricus, and G. quieta. Frogs<br />
become infected with H. varioplexus, a lung<br />
trematode, and H. eccentricus, a eustachian tube<br />
trematode, by eating infected odonates (Krull,<br />
1931; Dronen, 1975, 1978; Wetzel and Esch,<br />
1996). Glypthelmins quieta, a trematode of the<br />
small intestine, is acquired when frogs ingest<br />
prey such as tadpoles, frogs, and/or shed skin<br />
infected with metacercariae (Prudhoe and Bray,<br />
1982). Therefore, diet was important in the
transmission dynamics of these 3 trematode species<br />
in this study.<br />
Haematoloechus varioplexus and H. eccentricus<br />
were recovered from frogs throughout the<br />
year, increasing, although not significantly, in<br />
both prevalence and mean intensity during the<br />
fall collection. Recently, Wetzel and Esch (1997)<br />
have shown that the life span of H. eccentricus<br />
may be variable, with trematodes capable of maturing<br />
in as little as 1 wk and being lost the<br />
following week. Because of the small number of<br />
these flukes recovered in our study, little can be<br />
said about their recruitment throughout the year.<br />
Krull (1931) estimated that the life-span of Haematoloechus<br />
medioplexus Stafford, 1902, averaged<br />
1 yr, while studies by Kennedy (1980) on<br />
species of Haematoloechus have shown that<br />
trematodes can reach full length in only 60 days.<br />
The size differences observed for H. varioplexus<br />
during the year (Fig. 1) may be significant in<br />
understanding recruitment of this species. The<br />
seasonal variation in length of H. varioplexus<br />
suggests that adult worms are lost during early<br />
spring and that new infections begin during<br />
midspring and continue throughout the year.<br />
These results are similar to those of Ward<br />
(1909), who observed lung flukes of Rana pipiens<br />
Schreber, 1782, being lost during breeding,<br />
and recruitment occurring throughout the year.<br />
Two cestode species were recovered from<br />
green frogs during this study, the larval tetrathyridium<br />
of Mesocestoides sp. and a single<br />
specimen of an adult cestode that could not be<br />
identified because the scolex was lost. The complete<br />
life cycles of Mesocestoides spp. are currently<br />
unknown; however, a number of mammals,<br />
amphibians, and reptiles are known to<br />
serve as second intermediate hosts, while carnivorous<br />
mammals serve as definitive hosts. The<br />
tetrathyridian stage has been reported from a variety<br />
of mammals and reptiles but is rare in amphibians<br />
(McAllister and Conn, 1990). The life<br />
cycles of these 2 species of cestode are unknown,<br />
although frog diet may be important in<br />
their transmission dynamics.<br />
Nematodes in the genus Waltonella typically<br />
are found in the body cavity of species of Rana.<br />
Adult worms release microfilaria into the bloodstream,<br />
and mosquitoes serve as vectors, infecting<br />
frogs while feeding (Witenberg and Gerichter,<br />
1944). The only report of filarial worms in<br />
R. clamitans is of microfilaria recovered from 1<br />
frog in Ontario, Canada (Barta and Desser,<br />
BOLEK AND COGGINS—HELMINTH COMMUNITIES IN GREEN FROGS 169<br />
1984). Therefore, R. clamitans melanota is apparently<br />
a new host record for Waltonella sp.<br />
(Esslinger, 1986; Baker, 1987). Waltonella<br />
americana was previously reported and described<br />
in Wisconsin leopard frogs by Walton<br />
(1929).<br />
The nematode Cosmocercoides sp. was recovered<br />
from the large intestine of green frogs.<br />
Confusion exists in the literature on the identification<br />
of species of Cosmocercoides in amphibians<br />
and reptiles (Baker, 1987; Vanderburgh<br />
and Anderson, 1987). The major difference in<br />
species identification is the number of rosette<br />
papillae per subventral row in males, with male<br />
Cosmocercoides dukae Holl, 1928 (gastropod<br />
parasite) having 9—21 rosette papillae, averaging<br />
13-14, and C. variabilis (amphibian parasite)<br />
having 15-25, averaging 20 or 21. Because of<br />
this overlap and the presence of only 5 damaged<br />
males out of 62 nematodes recovered, species<br />
identification was not possible. Interestingly, no<br />
worms were found in the lungs or body cavity<br />
of any green frogs, and Cosmocercoides sp. occurred<br />
in frogs in months when gastropods were<br />
commonly found in the stomach contents. We<br />
suspect that specimens of Cosmocercoides sp.<br />
recovered are C. dukae, although this cannot be<br />
confirmed.<br />
Differential infection between host sex and<br />
prevalence or mean intensity was observed for<br />
a number of helminth species. Male frogs had a<br />
significantly higher prevalence of kidney metacercariae<br />
and significantly higher mean intensities<br />
of H. varioplexus and H. eccentricus than<br />
female frogs. Male green frogs are territorial<br />
during the breeding season and defend their<br />
aquatic breeding site from potential competitors<br />
(Martof, 1953; Oldham, 1967). Thus, unlike the<br />
females, they remain in the water for longer periods<br />
of time and may be exposed to cercariae<br />
of the kidney trematode for longer periods. Because<br />
males remain in a relatively small area of<br />
the pond during the breeding season, they may<br />
occur in a microhabitat conducive to becoming<br />
infected with digeneans. Recently, Wetzel and<br />
Esch (1997), in a seasonal study of Halipegus<br />
occidualis Stafford, 1905 and H. eccentricus in<br />
green frogs, suggested that certain areas of a<br />
pond may be "hot spots" for infection with digenetic<br />
trematodes. Therefore, male frogs in<br />
these "hot spots" may feed more often on<br />
emerging odonates containing metacercariae of<br />
species of Haematoloechus and Halipegus, ex-<br />
Copyright © 2011, The Helminthological Society of Washington
170 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
plaining the higher mean intensities of these<br />
trematodes observed in male frogs (Wetzel and<br />
Esch, 1996).<br />
Female frogs had significantly greater mean<br />
intensities of Cosrnocercoides sp. and Mesocestoides<br />
sp. than males. Although Cosmocercoides<br />
sp. could not be identified to species,<br />
both C. variabilis and C. dukae occur in terrestrial<br />
habitats. Female frogs spend more time<br />
on the ground and have a higher probability<br />
of encountering these nematodes in a terrestrial<br />
habitat, either by skin-penetrating C. variabilis<br />
or by feeding on terrestrial mollusks,<br />
hosts for C. dukae. Unfortunately, nothing can<br />
be stated about the transmission dynamics of<br />
Mesocestoides sp., and no conclusions can be<br />
drawn from this difference. The observed differences<br />
in host sex are probably due to ecological<br />
differences in their habitat preference<br />
throughout the year.<br />
Significant positive relationships between<br />
WW and species richness were observed in<br />
green frogs. In this study, frogs in the later<br />
collections had greater species richness than in<br />
early collections (Fig. 2); therefore, time of<br />
exposure was more important in developing<br />
richer helminth communities than was frog<br />
weight during the May-June, July-August,<br />
and September—October collections. This is<br />
supported by the results showing significant<br />
differences in species richness over time and<br />
nonsignificant correlations between WW and<br />
species richness. Observations linking higher<br />
species richness with larger host size have<br />
been reported in green frogs and other species<br />
of Rana by Muzzall (199 la) and Me Alpine<br />
(1997). These investigators suggested that older<br />
individuals may have a longer exposure<br />
time and possess more surface area for colonization<br />
by skin-penetrating nematodes and<br />
digenean metacercariae. Also, larger frogs<br />
possess a greater gape size and may feed on<br />
larger, and a wider number, of intermediate<br />
hosts than smaller individuals. As in their<br />
studies, our data also support the island size<br />
hypothesis, which predicts that larger host individuals<br />
should support higher species richness<br />
than smaller individuals (Holmes and<br />
Price, 1986). McAlpine (1997) also stated that<br />
aspects of host ecology, such as diet and habitat,<br />
and parasite transmission may confound<br />
any simple relationship between the diversity<br />
of helminth communities and size of hosts.<br />
Copyright © 2011, The Helminthological Society of Washington<br />
Data from the present study suggest that time<br />
of transmission may also have a similar confounding<br />
effect.<br />
The depauperate helminth community structure<br />
of Carroll <strong>College</strong> green frogs was similar<br />
to the community structure of green frogs examined<br />
from Michigan, U.S.A. and New<br />
Brunswick, Canada by Muzzall (199la) and<br />
McAlpine (1997), respectively. Of the 120<br />
frogs examined from Michigan, 108 (90%)<br />
were infected with a total of 13 species of helminths<br />
(8 trematodes, 1 cestode, and 4 nematodes)<br />
while 164 of 234 (70.1%) green frogs<br />
examined from Canada were infected with 18<br />
species of helminths (10 trematodes, 3 cestodes,<br />
and 5 nematodes). As in our study, in<br />
terms of abundance, digeneans dominated the<br />
adult helminth communities of frogs in Michigan<br />
(96.5%) and New Brunswick (60.8%), indicating<br />
that diet and a semiaquatic habitat<br />
were also important in structuring helminth<br />
communities dominated by indirect-life-cycle<br />
parasites at those locations. Similarly, in both<br />
of those studies, adult frogs had higher species<br />
richness than young-of-the-year individuals,<br />
indicating that size and/or age were also important<br />
in acquiring new species of helminths<br />
into the infracommunity of these hosts. Data<br />
from our study and the Michigan and New<br />
Brunswick frogs suggest that although helminth<br />
species composition, richness, and prevalence<br />
may be variable depending on collection<br />
site and the ecological factors influencing<br />
variation in life history traits in local populations<br />
at those locations, green frog helminth<br />
communities are dominated by digenetic trematodes<br />
acquired in a semiaquatic habitat and/<br />
or through the frogs' diet.<br />
Acknowledgments<br />
We thank Dr. Susan Lewis, Department of Biology,<br />
Can-oll <strong>College</strong> for allowing access to the<br />
field station and permission to collect frogs, and<br />
Melissa Ewert and Luke Bolek for help in collecting<br />
frogs. We also thank 2 anonymous reviewers<br />
and the editors Drs. Willis A. Reid, Jr.<br />
and Janet W Reid for improvements on an earlier<br />
draft of the manuscript.<br />
Literature Cited<br />
Aho, J. M. 1990. Helminth communities of amphibians<br />
and reptiles: comparative approaches to understanding<br />
patterns and processes. Pages 157-
196 in G. W. Esch, A. O. Bush, and J. W. Aho,<br />
eds. Parasite Communities: Patterns and Processes.<br />
Chapman and Hall, New York, New York,<br />
U.S.A. 335 pp.<br />
Baker, M. R. 1987. Synopsis of the Nematoda parasitic<br />
in amphibians and reptiles. Memorial University<br />
of Newfoundland Occasional Papers in Biology<br />
11:1-325.<br />
Barta, J. R., and S. S. Desser. 1984. Blood parasites<br />
of amphibians from Algonquin Park, Ontario.<br />
Journal of Wildlife Diseases 20:180-189.<br />
Barton, D. P. 1996. Helminth infracommunities in Litoria<br />
genimaculata (Amphibia: Anura) from<br />
Birthday Creek, an upland rainforest stream in<br />
northern Queensland, Australia. International<br />
Journal for <strong>Parasitology</strong> 26:381-385.<br />
Bolek, M. G., and J. R. Coggins. 2000. Seasonal occurrence<br />
and community structure of helminth<br />
parasites from the eastern American toad, Bufo<br />
americanus americanus, from southeastern Wisconsin,<br />
U.S.A. <strong>Comparative</strong> <strong>Parasitology</strong> 67:202-<br />
209.<br />
Borror, D. J., C. A. Triplehorn, and N. F. Johnson.<br />
1989. An Introduction to the Study of Insects, 6th<br />
ed. Harcourt Brace <strong>College</strong> Publishers, Philadelphia,<br />
Pennsylvania, U.S.A. 875 pp.<br />
Bush, A. O., K. D. Lafferty, J. M. Lotz, and A. W.<br />
Shostak. 1997. <strong>Parasitology</strong> meets ecology on its<br />
own terms: Margolis et al. revisited. Journal of<br />
<strong>Parasitology</strong> 83:575-583.<br />
Campbell, R. A. 19<strong>68</strong>. A comparative study of the<br />
parasites of certain Salientia from Pocahontas<br />
<strong>State</strong> Park, Virginia. Virginia Journal of Science<br />
19:13-29.<br />
Coggins, J. R., and R. A. Sajdak. 1982. A survey of<br />
helminth parasites in the salamanders and certain<br />
anurans from Wisconsin. Proceedings of the Helminthological<br />
Society of Washington 49:99-102.<br />
Dronen, N. O. 1975. Studies on the life cycle of Haematoloechus<br />
coloradensis Cort, 1915 (Digenea:<br />
Plagiorchiidae), with emphasis on host susceptibility<br />
to infection. Journal of <strong>Parasitology</strong> 61:657-<br />
660.<br />
. 1978. Host-parasite population dynamics of<br />
Haematoloechus coloradensis Cort, 1915 (Digenea:<br />
Plagiorchiidae). American Midland Naturalist<br />
99:330-349.<br />
Esslinger, J. H. 1986. Redescription of Foleyellid.es<br />
striatus (Ochoterena and Caballero, 1932) (Nematoda:<br />
Filarioidea) from a Mexican frog, Rana<br />
montezumae, with reinstatement of the genus Foleyellides<br />
Caballero, 1935. Proceedings of the Helminthological<br />
Society of Washington 53:218-223.<br />
Goater, T. M., G. W. Esch, and A. O. Bush. 1987.<br />
Helminth parasites of sympatric salamanders: ecological<br />
concepts at the infracommunity, component,<br />
and compound community levels. American<br />
Midland Naturalist 118:289-299.<br />
Goldberg, S. R., C. R. Bursey, and I. Ramos. 1995.<br />
The component parasite community of three sympatric<br />
toad species, Bufo cognatus, Bufo debilis<br />
(Bufonidae), and Spea multiplicata (Pelobatidae)<br />
from New Mexico. Journal of the Helminthological<br />
Society of Washington 62:57—61.<br />
BOLEK AND COGGINS—HELMINTH COMMUNITIES IN GREEN FROGS 171<br />
Hamilton, W. J., Jr. 1948. The food and feeding behavior<br />
of the green frog, Rana clamitans Latreille,<br />
in New York <strong>State</strong>. Copeia 1948:203-207.<br />
Holmes, J. C., and P. W. Price. 1986. Communities<br />
of parasites. Pages 187-213 in J. Kikkawa and D.<br />
J. Anderson, eds. Community Ecology: Pattern<br />
and Process. Blackwell Scientific Publications,<br />
Boston, Massachusetts, U.S.A. 432 pp.<br />
Kennedy, M. J. 1980. Host-induced variations in Haematoloechus<br />
buttensis (Trematoda: Haematoloechidae).<br />
Canadian Journal of Zoology 58:427-<br />
442.<br />
Kennedy, R. C., A. O. Bush, and J. M. Aho. 1986.<br />
Patterns in helminth communities: why are birds<br />
and fish different? <strong>Parasitology</strong> 93:205-215.<br />
Krull, W. H. 1931. Life history studies on two frog<br />
lung flukes, Pneumonoeces mediopiexus and<br />
Pneumobites parviplexus. Transactions of the<br />
American Microscopical Society 50:215—277.<br />
Martof, B. S. 1953. Territoriality in the green frog,<br />
Rana clamitans. Ecology 34:165-167.<br />
McAllister, C. T., and D. B. Conn. 1990. Occurrence<br />
of tetrathyridia of Mesocestoides sp. (Cestoidea:<br />
Cyclophyllidea) in North American anurans (Amphibia).<br />
Journal of Wildlife Diseases 26:540-543.<br />
McAlpine, D. F. 1997. Helminth communities in bullfrogs<br />
(Rana catesbeiand), green frogs (Rana<br />
clamitans), and leopard frogs (Rana pipiens) from<br />
New Brunswick, Canada. Canadian Journal of Zoology<br />
75:1883-1890.<br />
, and M. B. Burt. 1998. Helminths of bullfrogs,<br />
Rana catesbeiana, green frogs, R. clamitans,<br />
and leopard frogs, R. pipiens in New Brunswick.<br />
Canadian Field-Naturalist 112:50-<strong>68</strong>.<br />
-, and T. G. Dilworth. 1989. Microhabitat and<br />
prey size among three species of Rana (Anura:<br />
Ranidae) sympatric in eastern Canada. Canadian<br />
Journal of Zoology 67:2244-2252.<br />
Muzzall, P. M. 1991 a. Helminth infracommunities of<br />
the frogs Rana catesbeiana and Rana clamitans<br />
from Turkey Marsh, Michigan. Journal of <strong>Parasitology</strong><br />
77:366-371.<br />
. 1991b. Helminth communities of the newt,<br />
Notophthalrnus viridescens, from Turkey Marsh,<br />
Michigan. Journal of <strong>Parasitology</strong> 77:87-91.<br />
Oldham, R. S. 1967. Orienting mechanisms of the<br />
green frog, Rana clamitans. Ecology 48:477-491.<br />
Prudhoe, O. B. E., and R. A. Bray. 1982. Platyhelminth<br />
parasites of the Amphibia. British Museum<br />
(Natural History)/Oxford University Press, London,<br />
U.K. 217 pp.<br />
Scale, D. B. 1987. Amphibia. Pages 467-552 in T. J.<br />
Pandian and F. J. Vanberg, eds. Animal Energetics.<br />
Vol. 2. Academic Press, San Diego, California,<br />
U.S.A. 631 pp.<br />
Sokal, R. R., and J. F. Rohlf. 1981. Biometry, 2nd<br />
ed. W. H. Freeman and Company, New York, New<br />
York, U.S.A. 859 pp.<br />
Stewart, M. M., and P. Sandison. 1972. <strong>Comparative</strong><br />
food habits of sympatric mink frogs, bullfrogs and<br />
green frogs. Journal of Herpetology 6:241-244.<br />
Vanderburgh, D. J., and R. C. Anderson. 1987. The<br />
relationship between nematodes of the genus Cosmocercoides<br />
Wilkie, 1930 (Nematoda: Cosmocer-<br />
Copyright © 2011, The Helminthological Society of Washington
172 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
coidea) in toads (Bufo americanus) and slugs<br />
(Deroceras laeve). Canadian Journal of Zoology<br />
65:1650-1661.<br />
Vogt, R. C. 1981. Natural History of Amphibians and<br />
Reptiles of Wisconsin. The Milwaukee Public<br />
Museum and Friends of the Museum, Inc., Milwaukee,<br />
Wisconsin, U.S.A. 205 pp.<br />
Walton, A. C. 1929. Studies on some nematodes of<br />
North American frogs. I. Journal of <strong>Parasitology</strong><br />
15:227-249.<br />
Ward, H. B. 1909. The influence of hibernation and<br />
migration on animal parasites. Proceedings of the<br />
7th International Zoological Congress (Boston).<br />
12pp.<br />
Werner, E. E., G. A. Wellborn, and M. A. McPeek.<br />
1995. Diet composition in postmetamorphic bullfrogs<br />
and green frogs: implications for interspecific<br />
predation and competition. Journal of Herpetology<br />
29:600-607.<br />
Wetzel, E. J., and G. W. Esch. 1996. Influence of<br />
odonate intermediate host ecology on the infection<br />
dynamics of Halipegus spp., Haematoloechus longiplexus,<br />
and Haematoloechus complexus (Trematoda:<br />
Digenea). Journal of the Helminthological<br />
Society of Washington 63:1-7.<br />
-, and . 1997. Infrapopulation dynamics<br />
of Halipegus occidualls and Halipegus eccentricus<br />
(Digenea: Hemiuridae): temporal changes<br />
within individual hosts. Journal of <strong>Parasitology</strong><br />
83:1019-1024.<br />
Williams, D. D., and S. J. Taft. 1980. Helminths of<br />
anurans from NW Wisconsin. Proceedings of the<br />
Helminthological Society of Washington 47:278.<br />
Witenberg, G., and C. Gerichter. 1944. The mor<br />
phology and life history of Foleyella duboisis<br />
with remarks on allied filarids of amphibians.<br />
Journal of <strong>Parasitology</strong> 30:245-256.<br />
Yoder, H. R., and J. R. Coggins. 1996. Helminth<br />
communities in the northern spring peeper, Pseudacris<br />
c. crucifer Wied, and the wood frog, Rana<br />
sylvatica Le Conte, from southeastern Wisconsin.<br />
Journal of the Helminthological Society of Washington<br />
63:211-214.<br />
Announcement of the<br />
FOURTH INTERNATIONAL CONGRESS OF NEMATOLOGY<br />
at the<br />
Tenbel Resort, Tenerife, Canary Islands, Spain<br />
June 8-13, 2002<br />
Sponsor: The International Federation of Nematology Societies (IFNS).<br />
Scientific Program: 4 full days of scientific sessions with an opening plenary session, symposia, colloquia,<br />
discussion sessions and offered papers arranged in 4 poster sessions. The themes will include the systematics,<br />
molecular biology, genomics, genetics, management, ecology, and physiology of parasitic, entomophagous,<br />
and free-living nematodes. Abstracts of offered papers will be accepted between December<br />
1, <strong>2001</strong> and March 1, 2002.<br />
Accommodations: The Tenbel resort is a large hotel complex in an extensive tropical garden setting. A<br />
range of accommodations will be available to Congress participants. Special interest and scenic tours will<br />
be arranged during and after the FICN. Details of available accommodations and tourism opportunities<br />
will be posted on the IFNS web site at http:\\www.ifns.org.<br />
Registration: Registrations for the FICN will be accepted after December 1, <strong>2001</strong>. Registration forms<br />
and details regarding regular, student, spouse, and accompanying person registrations are available from<br />
the IFNS web site at www.ifns.org, or from Dr. Maria Arias, FICN Local Arrangements Chair, CSIC<br />
Centre de Ciencias Medioambiantales, Serrano 115 DPDO, Madrid 28006, Spain.<br />
Copyright © 2011, The Helminthological Society of Washington
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 173-176<br />
Blood Parasites of the Ring-Necked Duck (Ay thy a collaris} on Its<br />
Wintering Range in Florida, U.S.A.<br />
DONALD J. FORRESTER, 1
174 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Voucher specimens have been deposited in the International<br />
Reference Centre for Avian Haematozoa at<br />
the Queensland Museum, Brisbane, Australia (accession<br />
nos. G462509-G462640) and the U.S. National<br />
Parasite Collection in Beltsville, Maryland, U.S.A. (accession<br />
nos. 88248-88254). Prevalence data for each<br />
parasite were evaluated by using PROC FREQ for 2tailed<br />
Fisher's exact tests with regard to locality, gender,<br />
and age (SAS Institute, 1989). Significance was<br />
taken at P < 0.05. Terminology used was according to<br />
Bush et al. (1997).<br />
Results<br />
Five species of blood parasites were identified,<br />
2 protozoans (Haemoproteus nettionis<br />
(Johnson and Cleland, 1909) and Leucocytozoon<br />
simondi Mathis and Leger, 1910) and microfilariae<br />
of 3 nematodes (Splendidofilaria fallisensis<br />
(Anderson, 1954) and 2 unidentified species).<br />
The microfilariae of 5. fallisensis (n = 1,096)<br />
were 75.1 (41-130) in length and 4.9 (4.1-6.0)<br />
in width, with a rounded anterior, and a tapering<br />
tail that ended bluntly. Sheaths were lacking.<br />
Microfilariae of 2 unidentified filaroid species<br />
were observed in 36 ring-necked ducks. Microfilariae<br />
of Species I were long and slender with<br />
a bluntly rounded anterior and an overall uniform<br />
body width that tapered slightly to a rounded<br />
posterior. These microfilariae (n = 94) were<br />
150 (110-190) in length and 5.8 (5.0-6.0) in<br />
width. They had a well-defined sheath that could<br />
be seen at 1 or both ends of the body. The cephalic<br />
space was 4.8 (3-7, n = 10). Fixed points<br />
(n — 10), expressed as percentages of body<br />
length, were as follows: excretory cell 36.4 (35—<br />
38), inner body 64.9 (62-67), anal pore 86.6<br />
(83-90). Microfilariae of Species II (n = 287)<br />
were similar in appearance to Species I, except<br />
they were 150 (120-210) in length and 5.8 (5.0-<br />
6.0) in width and lacked a sheath. The cephalic<br />
space was 5.2 (3-6, n = 10). Fixed points (n =<br />
10) were as follows: excretory cell 36.5 (35-39),<br />
inner body 64.7 (63-67), anal pore 86.6 (85-<br />
90).<br />
The overall prevalences were H. nettionis<br />
(5.3%), L. simondi (9.2%), S. fallisensis<br />
(39.2%), filaroid Species I (6.0%), and filaroid<br />
Species II (13.8%). Of the sample, 97 ducks<br />
were infected with 1 species of parasite, 43 with<br />
2, and 8 with 3. The most common multiple infection<br />
was a combination of L. simondi and S.<br />
fallisensis (n = 15), followed by combinations<br />
of the 2 unidentified filaroids (n = 10).<br />
There were no significant differences in prevalences<br />
of the 5 species of parasites among the<br />
Copyright © 2011, The Helminthological Society of Washington<br />
collection periods, and therefore data for all<br />
years were combined. Prevalences of H. nettionis<br />
in Regions 1 (7%) and 2 (7%) were significantly<br />
higher (P = 0.03) than in Region 3 (0%).<br />
The prevalence of L. simondi was significantly<br />
higher (P = 0.0005) in Region 1 (19%) than in<br />
Regions 2 (8%) and 3 (1%). Microfilariae of<br />
Species II were more prevalent (P = 0.016) in<br />
Regions 2 (14%) and 3 (22%) than in Region 1<br />
(6%). There were no regional differences in the<br />
prevalences of the microfilariae of S. fallisensis<br />
and Species I.<br />
Microfilariae of S. fallisensis, Species I, and<br />
Species II were more prevalent in female ducks<br />
(46%, 9%, and 23%) than in males (33%, 3%,<br />
and 5%) (P = 0.028, 0.042, and 0.00002). The<br />
prevalences of H. nettionis and L. simondi were<br />
similar.<br />
Leucocytozoon simondi was more prevalent<br />
(P = 0.021) in juveniles (16%) than in adults<br />
(6%). On the other hand, filaroid Species I and<br />
II were both more prevalent (P = 0.029 and<br />
0.001) in adults (8% and 18%) than in juveniles<br />
(1% and 4%). There were no age differences in<br />
the prevalences of H. nettionis and S. fallisensis.<br />
Discussion<br />
Although this is the first report of H. nettionis<br />
and L. simondi from ring-necked ducks in Florida,<br />
both have been identified in ring-necked<br />
ducks overwintering in Texas, U.S.A. (Loven et<br />
al., 1980). Prevalences in the 20 ducks examined<br />
in Texas were similar to those of our Florida<br />
birds, i.e., 5% for H. nettionis and 10% for L.<br />
simondi. No microfilariae were reported in the<br />
Texas study. Unidentified microfilariae were<br />
seen in 1 of 6 ring-necked ducks examined in<br />
Maine, U.S.A.; the authors stated that their microfilariae<br />
were 45—65 long and 4-6 wide with<br />
a short, narrow, and slightly twisted tail (Nelson<br />
and Gashwiler, 1941). In a study of the blood<br />
parasites of 178 ring-necked ducks in the Maritime<br />
Provinces of Canada (New Brunswick,<br />
Nova Scotia, and Prince Edward Island), the<br />
prevalence of H. nettionis was higher (10%), and<br />
of L. simondi was lower (5%), than those in our<br />
birds from Florida (Bennett et al., 1975). No microfilariae<br />
were reported in the Maritime study.<br />
In another study, Bennett and Inder (1972)<br />
found microfilariae in 1 of 10 ring-necked ducks<br />
from Newfoundland, Canada, but provided no<br />
descriptions or measurements.<br />
Anderson's (1956) description of the micro-
filariae of S. fallisensis was similar to our specimens,<br />
with the exception of ranges of the<br />
lengths and the lack of sheaths; however, he noted<br />
that the sheaths of S. fallisensis were extremely<br />
delicate, and he was unable to see them<br />
in most specimens stained with Giemsa. Anderson<br />
(1956) also described a microfilaria from a<br />
European teal (Anas crecca Linnaeus, 1758) that<br />
he called Type D. His Type D microfilariae were<br />
similar to those of 5. fallisensis except for the<br />
length (range = 110-138) and the lack of a<br />
sheath. The microfilariae in our ring-necked<br />
ducks that we are calling 5. fallisensis could actually<br />
represent 2 species. However, it is possible<br />
that because we measured twice as many microfilariae<br />
as did Anderson (1956), that we have<br />
determined that the range of lengths for the microfilariae<br />
of this species is more extensive than<br />
previously recognized. Therefore, we are calling<br />
those microfilariae that fell in the range of 41-<br />
130 in length, but otherwise conformed to Anderson's<br />
(1956) description, S. fallisensis. The<br />
high number of combined infections of S. fallisensis<br />
and L. simondi in the same bird (n = 15)<br />
was probably due to the fact that both parasites<br />
utilize the same species of simuliid blackflies as<br />
vectors (Fallis et al., 1951; Anderson, 19<strong>68</strong>).<br />
Of the 36 ducks that had unidentified microfilariae,<br />
2 had Species I only, while 17 had Species<br />
II only. Seventeen of 36 ducks had both<br />
Species I and II microfilariae. The fact that Species<br />
I usually occurred with Species II and only<br />
twice by itself and that the range of lengths and<br />
several fixed points were almost equal may support<br />
the idea that these are variations of a single<br />
species. In many ways (morphologic and metric)<br />
our microfilariae resemble those of Chandlerella<br />
bus hi, described by Bartlett and Anderson<br />
(1987) from American coots (Fulica americana<br />
Gmelin, 1789) in Manitoba, Canada. They were<br />
not able to see the sheaths on microfilariae of C.<br />
bushi in blood films made from fresh heart blood<br />
and stained with Giemsa. However, they were<br />
able to see the sheaths on Giemsa-stained blood<br />
films of heart blood taken from thawed carcasses<br />
or specimens teased from lung tissue. The lack<br />
of visible sheaths of microfilariae in our ringnecked<br />
duck blood films might be because they<br />
were made from fresh heart blood. The identification<br />
of the microfilariae from ring-necked<br />
ducks will have to await the discovery of adult<br />
worms and further study and comparison of their<br />
FORRESTER ET AL.—BLOOD PARASITES OF RING-NECKED DUCKS 175<br />
intrauterine microfilariae with those from the<br />
blood.<br />
The regional differences in the prevalences of<br />
H. nettionis and L. simondi may have been a<br />
reflection of the location of the breeding grounds<br />
and flyways used by different subpopulations of<br />
ring-necked ducks. Because transmission of the<br />
blood parasites of waterfowl does not occur in<br />
Florida (Thul et al., 1980; Thul and O'Brien,<br />
1990; Forrester et al., 1994), the ducks must become<br />
infected either on the breeding grounds or<br />
during migration. The types and numbers of arthropod<br />
vectors found on various breeding<br />
grounds might differ and thereby influence the<br />
acquisition of these blood parasites in various<br />
segments of the North American population.<br />
Ring-necked ducks that overwinter in Florida<br />
are known to breed during the summer months<br />
in various prairie provinces of Canada across to<br />
Ontario and the eastern U.S.A. (Bellrose, 1976).<br />
Some ducks that breed in the more western regions<br />
of Canada migrate eastward and then<br />
move southward. Others migrate southward and<br />
pass through Wisconsin, Indiana, Tennessee, and<br />
Georgia. Most of the ring-necked ducks in Florida<br />
originate from Ontario, Manitoba, and the<br />
District of Mackenzie (Bellrose, 1976). The lower<br />
prevalence of infections of L. simondi in<br />
adults may be due to age-related immunity. Reasons<br />
for the gender differences in prevalences<br />
are unknown.<br />
Acknowledgments<br />
We thank H. F. Percival, T C. Hines, C. W.<br />
Jeske, and A. R. Woodward for assistance in collecting<br />
ducks, R. C. Littell for helping with statistical<br />
analyses, and R. C. Anderson, E. C. Greiner,<br />
and M. G. Spalding for reviewing the manuscript<br />
and offering useful suggestions. This research<br />
was supported in part by the Florida Fish<br />
and Wildlife Conservation Commission and is a<br />
contribution of Federal Aid to Wildlife Restoration,<br />
Florida Pitman—Robertson Project W-41.<br />
This is Florida Agricultural Experiment Station<br />
Journal Series No. R-07749.<br />
Literature Cited<br />
Anderson, R. C. 1956. Ornithofilaria fallisensis n. sp.<br />
(Nematoda: Filarioidea) from the domestic duck<br />
with descriptions of microfilariae in waterfowl.<br />
Canadian Journal of Zoology 32:125-137.<br />
. 19<strong>68</strong>. The simuliid vectors of Splendidofilaria<br />
fallisensis of ducks. Canadian Journal of Zoology<br />
46:610-611.<br />
Copyright © 2011, The Helminthological Society of Washington
176 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Bartlett, C. M., and R. C. Anderson. 1987. Chandlerella<br />
bushi n. sp. and Splendidofilaria caperata<br />
Hibler, 1964 (Nematoda: Filarioidea) from Fulica<br />
americana (Gruiformes: Rallidae) in Manitoba,<br />
Canada. Canadian Journal of Zoology 65:2799-<br />
2802.<br />
Bellrose, F. C. 1976. Ducks, Geese & Swans of North<br />
America. Stackpole Books, Harrisburg, Pennsylvania,<br />
U.S.A. 543 pp.<br />
Bennett, G. F., and J. G. Inder. 1972. Blood parasites<br />
of game birds from insular Newfoundland. Canadian<br />
Journal of Zoology 50:705-706.<br />
, A. D. Smith, W. Whitman, and M. Cameron.<br />
1975. Hematozoa of the Anatidae of the Atlantic<br />
Flyway. II. The Maritime Provinces of Canada.<br />
Journal of Wildlife Diseases 11:280-289.<br />
-, M. Whiteway, and C. B. Woodworth-Lynas.<br />
1982. Host-parasite catalogue of the avian<br />
Haematozoa. Memorial University of Newfoundland<br />
Occasional Papers in Biology 5:1-243.<br />
Bishop, M. A., and G. F. Bennett. 1992. Host-parasite<br />
catalogue of the avian Haematozoa, Supplement<br />
1, and Bibliography of the avian blood-inhabiting<br />
Haematozoa, Supplement 2. Memorial<br />
University of Newfoundland Occasional Papers in<br />
Biology 15:1-244.<br />
Bush, A. O., K. D. Lafferty, J. M. Lotz, and A. W.<br />
Shostak. 1997. <strong>Parasitology</strong> meets ecology on its<br />
own terms: Margolis et al. revisited. Journal of<br />
<strong>Parasitology</strong> 83:575-583.<br />
Fallis, A. M., D. M. Davies, and M. A. Vickers.<br />
1951. Life history of Leucocytozoon simondi<br />
Mathis and Leger in natural and experimental infections<br />
and blood changes produced in the avian<br />
host. Canadian Journal of Zoology 29:305-328.<br />
New Book Available<br />
Forrester, D. J., J. M. Kinsella, J. W. Mertins, R.<br />
D. Price, and R. E. Turnbull. 1994. Parasitic helminths<br />
and arthropods of fulvous whistling-ducks<br />
(Dendrocygna bicolor) in southern Florida. Journal<br />
of the Helminthological Society of Washington<br />
61:84-88.<br />
Loven, J. S., E. G. Bolen, and B. W. Cain. 1980.<br />
Blood parasitemia in a South Texas wintering waterfowl<br />
population. Journal of Wildlife Diseases<br />
16:25-28.<br />
Martin, E. M. 1996. Tables of 1981-1990 average<br />
U.S. waterfowl harvest by species and county.<br />
Unpublished data from the Division of Migratory<br />
Bird Management, U.S. Fish and Wildlife Service,<br />
Laurel, Maryland, U.S.A.<br />
Nelson, E. C., and J. S. Gashwiler. 1941. Blood parasites<br />
of some Maine waterfowl. Journal of Wildlife<br />
Management 5:199-205.<br />
Robertson, W. B., and G. E. Woolfenden. 1992.<br />
Florida Bird Species: An Annotated List. Florida<br />
Ornithological Society Special Publication Number<br />
6, Gainesville, Florida, U.S.A. 260 pp.<br />
SAS Institute, Inc. 1989. SAS/STAT User's Guide,<br />
Version 6, 4th ed., Vol. 1. SAS Institute, Inc.,<br />
Gary, North Carolina, U.S.A. 943 pp.<br />
Thul, J. E., D. J. Forrester, and E. C. Greiner. 1980.<br />
Hematozoa of Wood Ducks (Aix sponsd) in the<br />
Atlantic Flyway. Journal of Wildlife Diseases 16:<br />
383-389.<br />
, and T. O'Brien. 1990. Wood duck hematozoan<br />
parasites as biological tags: Development of<br />
a population assessment model. Pages 323-334 in<br />
Proceedings of the 1988 North American Wood<br />
Duck Symposium, St. Louis, Missouri, U.S.A.<br />
Metazoan Parasites in the Neotropics: A Systematic and Ecological Perspective. Editors Guillermo<br />
Salgado-Maldonado, Alfonso N. Garcia-Aldrete, and Victor M. Vidal-Martinez. 2000. Published by the<br />
Universidad Nacional Autonoma de Mexico (UNAM) to celebrate the 70th Anniversary of the Institute<br />
de Biologia. 310 pp. ISBN 9<strong>68</strong>-36-8827-6. 6%" X 8Vi" soft cover. Cost: US$20.00 or MXP$200.00 per<br />
copy plus shipping and handling. Available from Institute de Biologia, UNAM, Secretaria Tecnica, Apartado<br />
Postal 70-153, CP 04510 Mexico D.F., Mexico or on-line at the internet web-page http://<br />
www.ibiologia.unam.mx. Payment by check only, made payable to "Institute de Biologia, UNAM". This<br />
volume is a compilation of 11 original contributions by an internationally diverse group of 18 researchers<br />
addressing a broad range of parasite systematics and biogeographical issues in South and Central American<br />
vertebrates.<br />
Copyright © 2011, The Helminthological Society of Washington
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 177-184<br />
Parelaphostrongylus tennis (Nematoda: Protostrongylidae) and Other<br />
Parasites of White-Tailed Deer (Odocoileus virginianus} in Costa Rica<br />
RAMON A. CARRENO,1'5 LANCE A. DuRDEN,2 DANIEL R. BROOKS,3 ARTHUR ABRAMS,4 AND<br />
ERIC P. HOBERG4<br />
1 Department of Environmental Biology, University of Guelph, Guelph, Ontario, Canada NIG 2W1 (e-mail:<br />
rcarreno@uoguelph.ca), .<br />
2 Institute of Arthropodology and <strong>Parasitology</strong>, Georgia Southern University, <strong>State</strong>sboro, Georgia 30460,<br />
U.S.A. (e-mail: Idurden@gsvms2.cc.gasou.edu),<br />
3 Department of Zoology, University of Toronto, Toronto, Ontario, Canada M5S 3G5 (e-mail: dbrooks@<br />
zoo.utoronto.ca), and<br />
4 Biosystematics and National Parasite Collection Unit, USDA, Agricultural Research Service, BARC East<br />
1180, Beltsville, Maryland 20715, U.S.A.<br />
ABSTRACT: Parasites were collected from 2 female white-tailed deer (Odocoileus virginianus) in the Area de<br />
Conservacion Guanacaste, Costa Rica, in early June 1999. Both deer were parasitized by the ticks Amblyomma<br />
parvum and Haemaphysalis juxtakochi as well as the hippoboscid fly, Lipoptena mazamae. One deer also hosted<br />
the ticks Boophilus microplus, Ixodes qffinis, and Anocentor nitens. Both deer were infected by larvae of the<br />
nasopharyngeal botfly Cephenemyia jellisoni, and the helminths Eucyathostomum webbi, Gongylonema pulchrum,<br />
Parelaphostrongylus tennis, and Paramphistomum liorchis, whereas Setaria yehi, an undescribed species<br />
of Ashworthius, and Onchocerca cervipedis occurred in single hosts. A cysticercus of Taenia oinissa was found<br />
encapsulated in the lung parenchyma of 1 host. This is the first report of these endoparasites from Central<br />
America.<br />
KEY WORDS: Ashworthius sp., biodiversity, ticks, Boophilus microplus, Gongylonema pulchrum, Haemaphysalis<br />
juxtakochi, Ixodes affinis, Odocoileus virginianus, white-tailed deer, helminth parasites, Parelaphostrongylus<br />
tenuis, cysticercus, Costa Rica.<br />
The white-tailed deer Odocoileus virginianus<br />
(Zimmermann, 1780) has a widespread Nearctic<br />
and Neotropical range, extending from southern<br />
Canada and the United <strong>State</strong>s through Mexico<br />
and Central America to Bolivia, the Guianas,<br />
and northern Brazil (Reid, 1997). The subspecies<br />
described from Costa Rica, Odocoileus virginianus<br />
truei Merriam, 1898, ranges from the<br />
southeastern edge of Mexico to northeastern<br />
Panama (Whitehead, 1972; Mendez, 1984). The<br />
parasite fauna of O. virginianus and other cervids<br />
is well documented in North America<br />
(Walker and Becklund, 1970; Davidson et al.,<br />
1981). However, very little information is available<br />
on parasites of cervids in the southern parts<br />
of their range, including Central America. This<br />
is significant because white-tailed deer are hosts<br />
to several serious pathogens and parasites of cervids<br />
and other animals, including the tick Ixodes<br />
scapularis Say, 1821, which is the main North<br />
American vector of the agent of Lyme borreli-<br />
5 Corresponding author. Present address: Department<br />
of Nematology, University of California, Davis, California<br />
95616, U.S.A. (e-mail: racarreno@<br />
ucdavis.edu).<br />
177<br />
osis. Additionally, one of the most important<br />
parasites in O. virginianus is Parelaphostrongylus<br />
tenuis (Dougherty, 1945), the meningeal<br />
worm. This species is not pathogenic in O. virginianus,<br />
but when snails infected with its larvae<br />
are ingested by other ruminants such as moose<br />
(Alces alces (Linnaeus, 1758)), fallow deer<br />
(Dama dama (Linnaeus, 1758)), reindeer (Rangifer<br />
tarandus (Linnaeus, 1758)), and llamas<br />
(Lama spp.), severe neurologic disease can result<br />
from adult worms in the brain and central<br />
nervous system (Anderson, 1964, 1970; Nettles<br />
et al., 1977; Krogdahl et al., 1987; Rickard et<br />
al., 1994).<br />
The following report is part of a biodiversity<br />
inventory of eukaryotic parasites of vertebrates<br />
in the Area de Conservacion Guanacaste (ACG)<br />
in northwestern Costa Rica.<br />
Materials and Methods<br />
We collected 2 adult female O. virginianus within<br />
the ACG, Guanacaste, Costa Rica (10°57'N; 85°48'W)<br />
in early June 1999. Ectoparasites were collected within<br />
1 hr postmortem. Internal organs were then removed,<br />
following procedures suggested by Nettles (1981), and<br />
examined for endoparasites. In addition to onsite ex-<br />
Copyright © 2011, The Helminthological Society of Washington
178 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
amination, several aliquots of abomasal and intestinal<br />
contents were fixed and later searched for parasites.<br />
Contents from the abomasum and small intestine were<br />
suspended in 2 liter of water. Duplicate aliquots of 200<br />
ml were removed, fixed in 5% formalin, and subsequently<br />
examined using a stereomicroscope. Digeneans<br />
were fixed in hot 10% formalin and stored in 70%<br />
ethanol. Ectoparasites and nematodes were stored in<br />
70% ethanol without fixation in formalin. Prior to examination,<br />
nematodes were cleared in either glycerine<br />
or phenol-alcohol. Tissues examined for sarcocysts<br />
were embedded in paraffin, sectioned, and stained with<br />
both hematoxylin and eosin and periodic acid-Schiff<br />
stains for diagnosis. Amphistome digeneans were prepared<br />
using the same methods and identified using descriptions<br />
by Kennedy et al. (1985) and Sey (1991).<br />
Voucher tick specimens are deposited in the U.S. National<br />
Tick Collection (USNTC), Institute of Arthropodology<br />
and <strong>Parasitology</strong>, Georgia Southern University,<br />
<strong>State</strong>sboro, Georgia, U.S.A. Voucher specimens<br />
of other parasites are deposited in the U.S. National<br />
Parasite Collection (USNPC), United <strong>State</strong>s Department<br />
of Agriculture, Beltsville, Maryland, U.S.A. Accession<br />
numbers are listed in Table 1.<br />
Results<br />
A total of 18 parasite species was found in<br />
the 2 deer (Table 1). Both deer hosted the hippoboscid<br />
louse fly Lipoptena mazamae and the<br />
ixodid ticks Amblyomma parvum and Haemaphysalis<br />
juxtakochi. We also found second- and<br />
third-stage larvae of Cephenemyia jellisoni<br />
(identified using keys in Bennett and Sabrosky<br />
[1962]) in the nasal sinuses of both deer. One<br />
deer also hosted the ticks Anocentor nitens, Boophilus<br />
microplus, and Ixodes affinis.<br />
Both deer hosted the digenean trematode Paramphistomum<br />
liorchis in their rumens. They also<br />
were infected by 3 species of nematodes: Gongylonema<br />
pulchrum in the submucosa of the<br />
esophagus, Eucyathostomum webbi in the large<br />
intestine, and Parelaphostrongylus tennis in the<br />
inner surface of the dura and from cranial sinuses<br />
and nerves. Meristic data for the latter<br />
species (Table 2) did not differ from those reported<br />
for P. tennis in O. virginianus from North<br />
America (Anderson, 1963; Carreno and Lankester,<br />
1993), although the esophageal lengths of<br />
the 2 males were greater than those reported in<br />
North American specimens.<br />
A cysticercus of Taenia omissa was found encapsulated<br />
in fibrous connective tissue in the<br />
lung parenchyma. Identification of the cysticercus<br />
was based on structure and measurements of<br />
rostellar hooks and the occurrence in deer; morphology<br />
of the intact cysticercus was consistent<br />
with observations by Rausch (1981).<br />
Two specimens of Setaria yehi were collected<br />
from 1 deer, 1 in the rectum, and the other in<br />
the posterior region of the body cavity. From the<br />
abomasal intestinal aliquots, few nematodes in<br />
the abomasa and none in the small intestines<br />
were found. A female specimen of Mazamastrongylus<br />
sp. occurred in the abomasum of 1<br />
deer. It could not be identified to species based<br />
on the diagnostic features within the genus.<br />
Hoberg (1996) demonstrated polymorphism in<br />
vulvar anatomy, and no males were found. In<br />
the abomasum of the second deer, specimens of<br />
an undescribed species of Ashworthius were<br />
found.<br />
Protozoan infections were not obvious in<br />
these animals (blood smears and fecal examinations<br />
were negative), except for the presence<br />
of unidentified sarcocysts in the hind leg muscles<br />
of 1 host.<br />
Discussion<br />
Parasites in white-tailed deer<br />
Copyright © 2011, The Helminthological Society of Washington<br />
This report constitutes the first data on parasites<br />
of O. virginianus in Costa Rica. The ranges<br />
of C. jellisoni, P. liorchis, E. webbi, O. cervipedis,<br />
and, most importantly, P. tennis are now<br />
extended south of North America. These range<br />
extensions suggest that the parasites may also be<br />
present in Mexico, other parts of Central America,<br />
and perhaps South America, coinciding with<br />
the distribution of this cervid.<br />
There is little information on the parasites of<br />
cervids south of the United <strong>State</strong>s. Captive O.<br />
virginianus from the Yucatan Peninsula have<br />
been reported as hosts of nematodes (Haemonchus<br />
sp., Cooperia spp., Isospora spp., Eimeria<br />
spp., Trichuris spp., Strongyloides spp.) and cestodes<br />
(Moniezia spp.), based on examination of<br />
fecal samples and fecal culture of nematode L3<br />
larvae (Montes-Perez et al., 1998). Several parasites<br />
have been listed from O. virginianus from<br />
Mexico and Central America, including arthropods,<br />
Cephenemyia sp., L. mazamae, A. nitens,<br />
H. juxtakochi, I. affinis, and nematodes, Haemonchus<br />
conforms (Rudolphi, 1802) Cobb,<br />
1898, and Gongylonema pulchrum, as well as<br />
several other species not found in the present<br />
study (Mendez, 1984). In South America, O. virginianus<br />
has been reported as a host for the<br />
nematodes H. contortus, Setaria sp., Oesophagostomum<br />
asperum Railliet and Henry, 1913,<br />
and Mecistocirrus sp., as well as the arthropods
CARRENO ET AL.—DEER PARASITES IN COSTA RICA 179<br />
Table 1. Parasites recovered from Odocoileus virginianus in Guanacaste, Costa Rica.<br />
Protozoa<br />
Sarcocystis sp.<br />
Parasite Deer Deer 2 Accession number<br />
Arthropoda: Acari<br />
Arnblyomma parvum Aragao, 1908<br />
Anocentor nitens (Neumann, 1897)<br />
Boophilus microplus (Canestrini, 1887)<br />
Haemaphysalis juxtakochi (Cooley, 1946)<br />
Ixodes qffinis Neumann, 1899<br />
Amblyomma sp. (immature stages)<br />
Arthropoda: Diptera<br />
Lipoptena mazamae Rondani, 1878<br />
Cephenemyia jellisoni Townsend, 1941<br />
Trematoda<br />
Paramphistomum liorchis Fischoeder,<br />
1901<br />
Cestoda<br />
Taenia omissa Liihe, 1910<br />
Nematoda<br />
Setaria yehi (Desset, 1966)<br />
Ashworthius sp.<br />
Gongylonenm pulchmm Molin, 1857<br />
Eucyathostomum webbi Pursglove, 1976<br />
Parelaphostrongylus tennis (Dougherty,<br />
1945)<br />
Mazamastrongylus sp.<br />
Onchocerca cervipedis Wehr and Dikmans,<br />
1935<br />
+ USNPC 90056<br />
+ + RML 122837, RML 122838<br />
+ RML 122838<br />
+ - RML 122838<br />
+ + RML 122837, RML 122838<br />
+ - RML 122838<br />
+ + RML 122837, RML 122838<br />
+ USNPC 90062, USNPC 90063<br />
10 USNPC 90053, USNPC 90054<br />
+ (> 1,000) + (
180 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Table 2. Morphometric measurements for P. tennis<br />
recovered from O. virginianus in Guanacaste, Costa<br />
Rica; measurements are given in microns (u.m) unless<br />
otherwise noted.<br />
Males<br />
Length (mm)<br />
Width (posterior to esophagus)<br />
Esophagus length<br />
Nerve ring from anterior<br />
Excretory pore from anterior<br />
Gubernaculum length<br />
Crura length<br />
Spicules<br />
Spicule branch<br />
Females<br />
Length (mm)<br />
Width (posterior to esophagus)<br />
Nerve ring from anterior<br />
Excretory pore from anterior<br />
Vulva from posterior<br />
Anus from posterior<br />
N Mean Range<br />
1<br />
2<br />
2<br />
2<br />
1<br />
3<br />
3<br />
6<br />
2<br />
1<br />
2<br />
2<br />
2<br />
8<br />
8<br />
47.00<br />
170.00<br />
807.50<br />
120.00<br />
110.00<br />
117.00<br />
36.17<br />
225.00<br />
7 1 .50<br />
105.50<br />
210.00<br />
105.50<br />
108.50<br />
185.50<br />
56.96<br />
—<br />
140-200<br />
795-820<br />
107-133<br />
—<br />
110-131<br />
25.7-44.0<br />
211-236<br />
70-73<br />
—<br />
190-230<br />
102-109<br />
106-111<br />
165-214<br />
45.5-79.0<br />
gated geographically and seasonally. For example,<br />
in Alabama, Durden et al. (1991) recovered<br />
4 species of ticks from 537 O. virginianus examined<br />
during winter (November-January), but<br />
only 2 of these species (/. scapularis and Dermacentor<br />
albipictus (Packard, 1869)) were common.<br />
In Alberta, Samuel et al. (1980) recorded<br />
just 1 species of tick (D. albipictus) from 148<br />
O. virginianus examined in all months, whereas<br />
Smith (1977) recorded 7 tick species from this<br />
host from 12 southeastern states, again with just<br />
2 species (/. scapularis and Amblyomma americanum<br />
(Linnaeus, 1758)) being common. In<br />
southern Texas, Samuel and Trainer (1970) recovered<br />
6 species of ticks from 404 O. virginianus<br />
examined, with 3 of these species (A.<br />
americanum, Amblyomma inornatum Banks,<br />
1909, and Amblyomma maculatum Koch, 1844)<br />
being prevalent.<br />
The known geographical range of A. parvutn<br />
extends from Mexico to Argentina. Throughout<br />
this range it parasitizes a wide variety of mammals<br />
(Fairchild et al., 1966; Jones et al., 1972).<br />
In Panama, Fairchild et al. (1966) recorded A.<br />
parvum from white-tailed deer, cattle, domestic<br />
cats, sloth, human, anteater (Tamandua sp.), and<br />
cotton rats (Sigmodon spp.). There is an unpublished<br />
USNTC record of this tick from a horse<br />
in Costa Rica.<br />
The tropical horse tick, A. nitens, is a pest of<br />
Copyright © 2011, The Helminthological Society of Washington<br />
equines in the neotropics and is the main vector<br />
of Babesia equi (Laveran, 1901), the protozoan<br />
that causes equine piroplasmosis (Strickland et<br />
al., 1976). In adjoining Panama, Fairchild et al.<br />
(1966) reported this tick from horses, cattle, and<br />
deer. The USNTC contains 8 collections of A.<br />
nitens from Costa Rica: 6 from horses, 1 from<br />
a domestic cat, and 1 from a rabbit (Sylvilagus<br />
sp.).<br />
The southern cattle tick, B. microplus, was<br />
formerly widespread throughout the New World<br />
as a major pest of domestic cattle, where it<br />
caused tremendous economic damage through<br />
its role as a vector of Babesia bigemina (Smith<br />
and Kilborne, 1893), an agent of bovine piroplasmosis<br />
(=Texas cattle fever) (Strickland et<br />
al., 1976; Bram and George, 2000). This tick<br />
also is a vector of Anaplasma marginale Theiler,<br />
1910 and Babesia bovis (Babes, 1888) Starcovici,<br />
1893. In Costa Rica, Hermans et al. (1994)<br />
reported high infection rates for these 3 hemoparasites<br />
in B. microplus and high seroprevalences<br />
to them in cattle. In Panama, Fairchild et<br />
al. (1966) reported this tick from cattle, horses,<br />
pigs, and dogs, with single collections from a<br />
goat and a deer. The USNTC contains 23 collections<br />
of B. microplus from Costa Rica: 15<br />
from cattle, 2 from horses, 2 from vegetation,<br />
and 1 each from a human, a tapir (Tapirus sp.),<br />
a gray fox (Urocyon cinereoargenteus (Schreber,<br />
1775)), and a fringe-lipped bat (Trachops<br />
cirrhosus (Spix, 1823)).<br />
Deer are the preferred hosts of H. juxtakochi,<br />
which is widely distributed from Mexico to Argentina<br />
(Jones et al., 1972). However, this tick<br />
has occasionally also been collected from rodents,<br />
humans, tapirs, coatimundis (Nasua sp.),<br />
peccaries (Tayassu spp.), porcupines (Coendou<br />
spp.), and lagomorphs (Fairchild et al., 1966;<br />
Jones et al., 1972). The USNTC contains 1 other<br />
Costa Rican collection of H. juxtakochi, also<br />
from a white-tailed deer.<br />
Ixodes affinis has a disjunct geographical distribution,<br />
with 1 focus in the southeastern<br />
U.S.A. (coastal Florida, Georgia, and South Carolina),<br />
and the other focus extending from Mexico<br />
to Brazil (Fairchild et al., 1966; Durden and<br />
Keirans, 1996). Because it is a member of the<br />
Ixodes ricinus complex, several members of<br />
which are vectors of the Lyme disease spirochete,<br />
Borrelia burgdorferi Johnson, Schmid,<br />
Hyde et al., 1984, this tick could be an enzootic<br />
vector of this zoonotic pathogen. It parasitizes a
ange of host species, but most collections, especially<br />
of adults, are from deer and larger carnivores<br />
(Fairchild et al., 1966; Durden and Keirans,<br />
1996). The USNTC includes 5 additional<br />
Costa Rican collections of /. qffinis: 2 from ocelots<br />
(Leopardus pardalis (Linnaeus, 1758)), and<br />
1 each from a human, a horse, and a long-tailed<br />
weasel (Mustela frenata Lichtenstein, 1831).<br />
Digeneans<br />
Brokx (1984) and Mendez (1984) reported<br />
amphistome digeneans, Cotylophoron sp. and<br />
Paramphistomum cervi (Zeder, 1790), respectively,<br />
in the stomach of O. virginianus. Unfortunately<br />
no voucher specimens exist from those<br />
accounts, so we cannot confirm their identifications.<br />
The only amphistomes we found were P.<br />
liorchis, the species most commonly reported<br />
from O. virginianus in North America (Kennedy<br />
et al., 1985).<br />
Cestodes<br />
Taenia omissa has a broad geographic distribution<br />
in the Western Hemisphere, coinciding<br />
with the range of the cougar, Puma concolor<br />
(Linnaeus, 1771), and deer intermediate hosts<br />
including Odocoileus and Mazama in North and<br />
South America (Rausch, 1981; Rausch et al.,<br />
1983). Consistent with the current study, cysticerci<br />
generally are found in the thoracic cavity,<br />
including the lungs and pericardium (Forrester<br />
and Rausch, 1990). Although cysticerci have<br />
been reported in brocket deer (Mazama cf. gouazoubira<br />
(Fischer, 1814)) from eastern Colombia<br />
(Rausch, 1981), there are apparently no prior records<br />
from Central America. Prevalence and intensity<br />
of infection in deer may be influenced by<br />
differences in population density of cougars<br />
across the range of this parasite—host assemblage<br />
(Forrester and Rausch, 1990).<br />
Nematodes<br />
This is the first record of P. tennis south of<br />
the United <strong>State</strong>s. The presence of elaphostrongyline<br />
nematodes in cervids is a major concern<br />
in the translocation of these animals in wildlife<br />
projects and the game ranching industry (Lankester<br />
and Fong, 1989; Samuel et al., 1992;<br />
Miller and Thorne, 1993; Davidson et al., 1996).<br />
Parelaphostrongylus tennis is of great concern<br />
in future wildlife management and conservation<br />
practices in Central America. An overall decline<br />
of O. virginianus populations in Mexico and<br />
CARRENO ET AL.—DEER PARASITES IN COSTA RICA 181<br />
Central America due to overhunting and habitat<br />
loss (Mendez, 1984) raises the possibility of reintroducing<br />
deer to areas where they have been<br />
extirpated. The effects of P. tennis on the only<br />
other Central American cervid, the brocket deer<br />
(Mazama americana (Erxleben, 1777)), are unknown.<br />
As P. tennis is highly pathogenic in<br />
most cervids other than O. virginianus, it may<br />
also be pathogenic in Mazama spp. Until the<br />
pathogenic significance (if any) of P. tennis to<br />
M. americana has been determined, the translocation<br />
of both it and Central American O. virginianus<br />
may be problematic in areas inhabited<br />
by other cervids that may be susceptible to parelaphostrongylosis.<br />
The translocation of infected O. virginianus<br />
to areas in which P. tennis is absent may result<br />
in the establishment of the parasite in other areas.<br />
The importation of deer from Pennsylvania<br />
to an island off the Georgia coast may have resulted<br />
in the establishment of P. tennis in an area<br />
outside its normal range (Davidson et al., 1996).<br />
Similarly, the translocation of reindeer (Rangifer<br />
tarandus) from Norway to Newfoundland has<br />
led to the establishment of Elaphostrongylus<br />
rangiferi Mitskevitch, 1960, another pathogenic<br />
species, in this region of North America (Lankester<br />
and Fong, 1989). Other cervids such as<br />
red deer (Cervus elaphus Linnaeus, 1758) and<br />
moose (Alces alces (Linnaeus, 1758)) may also<br />
harbor Elaphostrongylus species, and North<br />
American elk have been shown to have potential<br />
for surviving infection with and passing larvae<br />
of P. tennis (Samuel et al., 1992). These studies<br />
indicate a need for reliable diagnosis of P. tennis<br />
in ungulates and the need to determine the full<br />
distribution of this parasite. This information can<br />
help to prevent the spread of the parasite to uninfected<br />
host populations. Cervids of western<br />
North America are of particular concern, as P.<br />
tennis has not been recorded in western states<br />
and provinces (Miller and Thorne, 1993).<br />
The presence of P. tennis in Central American<br />
deer is also of evolutionary significance. As yet,<br />
we do not know if M. americana or any of the<br />
South American cervids such as Pudu spp. and<br />
Ozotoceros spp. are hosts for protostrongylids.<br />
Phylogenetic analysis of the family Protostrongylidae<br />
and comparison with host distribution,<br />
however, indicates that cervids are the basal<br />
hosts of these parasites (Carreno and Hoberg,<br />
1999). Discovery of new or already described<br />
protostrongyles in these hosts will contribute<br />
Copyright © 2011, The Helminthological Society of Washington
182 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
further to an understanding of the evolution of<br />
the Protostrongylidae.<br />
The presence of an undescribed species of<br />
Ashworthius is also of interest. Species of this<br />
genus of the Haemonchiinae have never been<br />
reported from hosts in the Western Hemisphere,<br />
although both Haernonchus and Mecistocirrus<br />
are known in Central and South America. In Africa<br />
and Eurasia, species of Ashworthius have<br />
been reported from cervids and members of 2<br />
subfamilies of the Bovidae, but not species of<br />
Bos (Pike, 1969; Drozdz et al., 1998). The presence<br />
of Ashworthius sp. in a wild cervid in the<br />
Western Hemisphere may have been the result<br />
of introduction and colonization from bovine<br />
hosts following European settlement since the<br />
1500s (Hoberg, 1997). The host distribution for<br />
this genus and its apparent absence in domestic<br />
bovids, however, suggest otherwise. Alternatively,<br />
the distribution of Ashworthius spp. in Central<br />
America may be relatively archaic, reflecting<br />
an extended history with endemic cervids. Historical<br />
reconstruction of the biogeography of<br />
Ashworthius in the New World is currently hampered<br />
by the paucity of survey data regarding<br />
parasitism in wild ruminants in Central and<br />
South America. Additionally, it is important to<br />
note that superficially some species of Ashworthius<br />
may resemble and be confused with Haemonchus<br />
spp.<br />
Conclusions<br />
The purpose of parasite inventories is 2-fold.<br />
First, the information obtained from parasite<br />
faunas can contribute valuable integrative information<br />
on our knowledge of the biosphere that<br />
serves as an indicator of biodiversity (Brooks<br />
and Hoberg, 2000). Secondly, in the case of the<br />
white-tailed deer, it is important to develop an<br />
understanding of the distribution of potential<br />
pathogens of wildlife. Many wildlife pathogens<br />
pose a serious threat to global biodiversity and<br />
also include several zoonoses (Daszak et al.,<br />
2000), and it is thus necessary to assess carefully<br />
the global distribution of potentially pathogenic<br />
parasites (Hoberg, 1997). The results of this<br />
study are an important contribution to biodiversity<br />
initiatives in Costa Rica. They have important<br />
implications in conservation projects in the<br />
region, as some parasites of white-tailed deer,<br />
such as P. tennis, may be pathogenic in other,<br />
less common endemic hosts such as M. arneri-<br />
Copyright © 2011, The Helminthological Society of Washington<br />
The uncertainty of species identifications in<br />
other reports, such as the amphistome digeneans<br />
(Brokx, 1984; Mendez, 1984), demonstrates the<br />
need for deposition of suitable voucher specimens<br />
in documenting parasite fauna. Are we<br />
dealing with a widespread and relatively uniform<br />
parasite fauna of white-tailed deer throughout<br />
their range, or one that is highly localized<br />
depending on habitat? A lack of voucher specimens<br />
to confirm identifications, and a lack of<br />
sufficient expert taxonomists available to provide<br />
those identifications (the taxonomic impediment:<br />
Brooks and Hoberg, 2000), prevents us<br />
from making such assessments. And such assessments,<br />
in turn, are critical for management<br />
policy, including game farming, sport hunting,<br />
and the interface between wildlands and agroscape.<br />
Acknowledgments<br />
We thank the scientific and technical staff of<br />
the ACG for support of this study, in particular<br />
Sigifredo Marin, Roger Blanco, Alejandro Masis,<br />
Maria Marta Chavarria, Felipe Chavarria,<br />
Guillermo Jimenez, Carolina Cano, Elda Araya,<br />
Fredy Quesada, Dunia Garcia, Roberto Espinoza,<br />
Elba Lopez, and Petrona Rios. Special thanks<br />
to Dr. Dan Janzen, technical adviser to the ACG,<br />
and to Calixto Moraga for their timely and accurate<br />
assistance. We are also grateful to Dr.<br />
Murray Lankester, Lakehead University, Thunder<br />
Bay, Canada, for advice on necropsy and<br />
collection procedures. Funds for this study were<br />
provided in part by an operating grant from the<br />
Natural Sciences and Engineering Research<br />
Council (NSERC) of Canada to D.R.B. Study of<br />
tick specimens was funded in part by National<br />
Institutes of Health grant Al 40729 to L.A.D.<br />
Literature Cited<br />
Anderson, R. C. 1963. The incidence, development,<br />
and experimental transmission of Pneiunostrongylus<br />
tenuis Dougherty (Metastrongyloidea: Protostrongylidae)<br />
of the meninges of the white-tailed<br />
deer (Odocoileus virginianus borealis) in Ontario.<br />
Canadian Journal of Zoology 41:775-792.<br />
. 1964. Neurologic disease in moose infected<br />
experimentally with Pneiimostrongyliis tennis<br />
from white-tailed deer. Pathologia Veterinaria 1:<br />
289-322.<br />
-. 1970. Neurologic disease in reindeer (Rangifer<br />
tarandus tarandus) introduced into Ontario.<br />
Canadian Journal of Zoology 49:159-166.<br />
Bennett, G. F., and C. W. Sabrosky. 1962. The Ne<br />
arctic species of the genus Cephenemyla (Diptera,
Oestridae). Canadian Journal of Zoology 40:431-<br />
448.<br />
Bram, R. A., and J. E. George. 2000. Introduction<br />
of nonindigenous arthropod pests of animals.<br />
Journal of Medical Entomology 37:1-8.<br />
Brokx, P. A. 1984. South America. Pages 525-546 in<br />
L. K. Halls, ed. White-Tailed Deer: Ecology and<br />
Management. Wildlife Management Institute.<br />
Stackpole Books, Harrisburg, Pennsylvania,<br />
U.S.A. 870 pp.<br />
Brooks, D. R., and E. P. Hoberg. 2000. Triage for<br />
the biosphere: the need and rationale for taxonomic<br />
inventories and phylogenetic studies of parasites.<br />
<strong>Comparative</strong> <strong>Parasitology</strong> 67:1—25.<br />
Capelle, K. J. 1971. Myiasis. Pages 279-305 in J. W.<br />
Davis and R. C. Anderson, eds. Parasitic Diseases<br />
of Wild Mammals. Iowa <strong>State</strong> University Press,<br />
Ames, Iowa, U.S.A.<br />
Carreno, R. A., and E. P. Hoberg. 1999. Evolutionary<br />
relationships among the Protostrongylidae<br />
(Nematoda: Metastrongyloidea) as inferred from<br />
morphological characters, with consideration of<br />
parasite—host coevolution. Journal of <strong>Parasitology</strong><br />
85:638-648.<br />
, and M. W. Lankester. 1993. Additional information<br />
on the morphology of the Elaphostrongylinae<br />
(Nematoda: Protostrongylidae) of North<br />
American Cervidae. Canadian Journal of Zoology<br />
71:592-600.<br />
Daszak, P., A. A. Cunningham, and A. D. Hyatt.<br />
2000. Emerging infectious diseases of wildlife—<br />
threats to biodiversity and human health. Science<br />
287:443-449.<br />
Davidson, W. R., G. L. Doster, and R. C. Freeman.<br />
1996. Parelaphostrongylus tennis on Wassaw Island,<br />
Georgia: a result of translocating whitetailed<br />
deer. Journal of Wildlife Diseases 32:701-<br />
703.<br />
, F. A. Hayes, V. F. Nettles, and F. E. Kellogg<br />
(eds.). 1981. Diseases and Parasites of White-<br />
Tailed Deer. Miscellaneous Publication No. 7, Tall<br />
Timbers Research Station, Tallahassee, Florida,<br />
U.S.A. 458 pp.<br />
Drozdz, J., A. W. Demiaszkiewicz, and J. Lachowicz.<br />
1998. Ashworthius sidemi (Nematoda, Trichostrongylidae)<br />
a new parasite of the European bison<br />
Bison bonasus (L.) and the question of independence<br />
of A. gagarini. Acta Parasitologica 43:75-<br />
80.<br />
Durden, L. A., and J. E. Keirans. 1996. Nymphs of<br />
the genus Ixodes (Acari: Ixodidae) of the United<br />
<strong>State</strong>s: taxonomy, identification key, distribution,<br />
hosts, and medical/veterinary importance. Thomas<br />
Say Publications in Entomology: Monographs,<br />
Entomological Society of America, Lanham,<br />
Maryland, U.S.A. 95 pp.<br />
, S. Luckhart, G. R. Mullen, and S. Smith.<br />
1991. Tick infestations of white-tailed deer in Alabama.<br />
Journal of Wildlife Diseases 27:606-614.<br />
Fairchild, G. B., G. M. Kohls, and V. J. Tipton.<br />
1966. The ticks of Panama (Acarina: Ixodoidea).<br />
Pages 167-219 in R. L. Wenzel and V. J. Tipton,<br />
eds. Ectoparasites of Panama. Field Museum of<br />
Natural History, Chicago, Illinois, U.S.A.<br />
CARRENO ET AL.—DEER PARASITES IN COSTA RICA 183<br />
Forrester, D. J., and R. L. Rausch. 1990. Cysticerci<br />
(Cestoda: Taeniidae) from white-tailed deer, Odocoileus<br />
virginianus, in southern Florida. Journal of<br />
<strong>Parasitology</strong> 76:583-585.<br />
Hermans, P., R. H. Dwinger, G. M. Buening, and<br />
M. V. Herrero. 1994. Seasonal incidence and hemoparasite<br />
infection rates of ixodid ticks (Acari,<br />
Ixodidae) detached from cattle in Costa Rica. Revista<br />
de Biologia Tropical 42:623-632.<br />
Hoberg, E. P. 1996. Emended description of Mazamastrongylus<br />
peruvianus (Nematoda: Trichostrongylidae),<br />
with comments on the relationships of<br />
the genera Mazamastrongylus and Spiculopteragia.<br />
Journal of <strong>Parasitology</strong> 82:470-477.<br />
. 1997. Parasite biodiversity and emerging<br />
pathogens: a role for systematics in limiting impacts<br />
on genetic resources. Pages 71-83 in K. E.<br />
Hoagland and A. Y. Rossman, eds. Global Genetic<br />
Resources: Access, Ownership and Intellectual<br />
Property Rights. Association of Systematics Collections,<br />
Washington, D.C., U.S.A.<br />
Jones, E. K., C. M. Clifford, J. E. Keirans, and G.<br />
M. Kohls. 1972. The ticks of Venezuela (Acarina:<br />
Ixodoidea) with a key to the species of Amblyornma<br />
in the western hemisphere. Brigham Young<br />
University Science Bulletin, Biological Series<br />
17(4): 1-40.<br />
Kennedy, M. J., M. W. Lankester, and J. B. Snider.<br />
1985. Paramphistomum cervi and Paramphistomum<br />
liorchis (Digenea: Paramphistomatidae) in<br />
moose, Alces alces, from Ontario, Canada. Canadian<br />
Journal of Zoology 63:1207-1210.<br />
Krogdahl, D. W., J. P. Thilstead, and S. K. Olsen.<br />
1987. Ataxia and hypermetria caused by Parelaphostrongylus<br />
tennis infection in llamas. Journal<br />
of the American Veterinary Medical Association<br />
190:191-193.<br />
Lankester, M. W., and D. Fong. 1989. Distribution<br />
of elaphostrongyline nematodes (Metastrongyloidea:<br />
Protostrongylidae) in Cervidae and possible<br />
effects of moving Rangifer spp. into and within<br />
North America. Alces 25:133-145.<br />
Maa, T. C. 1963. Genera and species of Hippoboscidae<br />
(Diptera): types, synonymy, habitats and natural<br />
groupings. Pacific Insects Monograph 6:1-<br />
186.<br />
Mendez, E. 1984. Mexico and Central America. Pages<br />
513-524 in L. K. Halls, ed. White-Tailed Deer:<br />
Ecology and Management. Wildlife Management<br />
Institute. Stackpole Books, Harrisburg, Pennsylvania,<br />
U.S.A. 870 pp.<br />
Miller, M. W., and E. T. Thorne. 1993. Captive cervids<br />
as potential sources of disease for North<br />
America's wild cervid populations: avenues, implications,<br />
and preventive management. Transactions<br />
of the North American Wildlife and Natural<br />
Resources Conference 58:460-467.<br />
Montes-Perez, R. C., R. I. Rodriguez-Vivas, J. F. de<br />
J. Torres-Acosta, and L. G. Ek-Pech. 1998. Seguimiento<br />
anual de la parasitosis gastrointestinal<br />
de venados cola blanca Odocoileus virginianus<br />
(Artiodactyla: Cervidae) en cautiverio en Yucatan,<br />
Mexico. Revista de Biologia Tropical 46:821-827.<br />
Nettles, V. F. 1981. Necropsy procedures. Pages 6-16<br />
Copyright © 2011, The Helminthological Society of Washington
184 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
in W. R. Davidson, F. A. Hayes, V. F. Nettles, and<br />
F. E. Kellogg, eds. Diseases and Parasites of<br />
White-Tailed Deer. Miscellaneous Publication no.<br />
7, Tall Timbers Research Station. Southeastern<br />
Cooperative Wildlife Disease Study, Athens,<br />
Georgia, U.S.A. 458 pp.<br />
, A. K. Prestwood, and R. D. Smith. 1977.<br />
Cerebrospinal parelaphostrongylosis in fallow<br />
deer. Journal of Wildlife Diseases 13:440-444.<br />
Pike, A. 1969. A revision of the genus Ashworthius<br />
Le Roux, 1930 (Nematoda: Trichostrongylidae).<br />
Journal of Helminthology 43:135-144.<br />
Rausch, R. L. 1981. Morphological and biological<br />
characteristics of Taenia rileyi Loewen, 1929<br />
(Cestoda: Taeniidae). Canadian Journal of Zoology<br />
59:653-666.<br />
, C. Maser, and E. P. Hoberg. 1983. Gastrointestinal<br />
helminths of the cougar, Felis concolor<br />
L., in northeastern Oregon. Journal of Wildlife<br />
Diseases 19:14-19.<br />
Reid, F. A. 1997. A Field Guide to the Mammals of<br />
Central America and Southeast Mexico. Oxford<br />
University Press, Oxford, U.K. 334 pp.<br />
Rickard, L. G., B. B. Smith, E. J. Gentz, A. A.<br />
Frank, E. G. Pearson, L. L. Walker, and M. J.<br />
Pybus. 1994. Experimentally induced meningeal<br />
worm (Parelaphostrongylus tennis) infection in<br />
the llama (Lama glama): clinical evaluation and<br />
implications for parasite translocation. Journal of<br />
Zoo and Wildlife Medicine 25:390-402.<br />
Samuel, W. M., E. R. Grinnell, and A. J. Kennedy.<br />
1980. Ectoparasites (Mallophaga, Anoplura, Acari)<br />
on mule deer, Odocoileus hemionus, and whitetailed<br />
deer, Odocoileus virginianus, of Alberta,<br />
Canada. Journal of Medical Entomology 17:15-<br />
17.<br />
, M. J. Pybus, D. A. Welch, and C. J. Wilke.<br />
1992. Elk as a potential host for meningeal worm:<br />
Copyright © 2011, The Helminthological Society of Washington<br />
implications for translocation. Journal of Wildlife<br />
Management 56:629-639.<br />
-, and D. O. Trainer. 1970. Amblyomma (Ac-<br />
arina: Ixodidae) on white-tailed deer, Odocoileus<br />
virginianus (Zimmermann), from South Texas<br />
with implications for theileriasis. Journal of Medical<br />
Entomology 7:567-574.<br />
Sey, O. 1991. CRC Handbook of the Zoology of Amphistomes.<br />
CRC Press Inc., Boca Raton, Florida,<br />
U.S.A. 480 pp.<br />
Smith, J. S. 1977. A survey of ticks infesting whitetailed<br />
deer in 12 southeastern states. M.S. Thesis,<br />
University of Georgia, Athens, Georgia, U.S.A. 61<br />
pp.<br />
Strickland, R. K., R. R. Gerrish, J. L. Hourrigan,<br />
and G. O. Schubert. 1976. Ticks of Veterinary<br />
Importance. USD A, APHIS Handbook No. 485.<br />
U.S. Government Printing Office, Washington.<br />
D.C., U.S.A. 122 pp.<br />
, , and J. S. Smith. 1981. Arthropods.<br />
Pages 363-389 in W. R. Davidson, F. A. Hayes,<br />
V. F. Nettles, and F. E. Kellogg, eds. Diseases and<br />
Parasites of White-Tailed deer. Miscellaneous<br />
Publication no. 7, Tall Timbers Research Station.<br />
Southeastern Cooperative Wildlife Disease Study,<br />
Athens, Georgia, U.S.A. 458 pp.<br />
Tonn, R. J., G. M. Kohls, and K. Arnold. 1963.<br />
Ectoparasites of birds and mammals of Costa<br />
Rica. 2. Ticks. Revista de Biologia Tropical 11:<br />
217-220.<br />
Walker, M. L., and W. W. Becklund. 1970. Checklist<br />
of the internal and external parasites of deer,<br />
Odocoileus hemionus and O. virginianus, in the<br />
United <strong>State</strong>s and Canada. Special Publication No.<br />
1, Index Catalogue of Medical and Veterinary Zoology,<br />
U.S. Government Printing Office, Washington,<br />
D.C., U.S.A. 45 pp.<br />
Whitehead, G. K. 1972. Deer of the World. Constable<br />
and Company, London, U.K. 194 pp.
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 185-189<br />
Cepedietta michiganensis (Protozoa) and Batracholandros<br />
magnavulvaris (Nematoda) from Plethodontid Salamanders in West<br />
Virginia, U.S.A.<br />
JAMES E. JOY' AND ROBERT B. TUCKER<br />
Department of Biological Sciences, Marshall University, Huntington, West Virginia 25755,<br />
U.S.A. (e-mail: joy@marshall.edu)<br />
ABSTRACT: The gastrointestinal tracts of 38 plethodontid salamanders (25 Plethodon punctatus and 13 Plethodon<br />
wehrlei), collected at high elevation sites in Pendleton and Randolph counties, West Virginia, U.S.A., were<br />
examined for parasites in 1996. Sixty percent of P. punctatus and 23% of P. wehrlei were infected by the ciliate<br />
Cepedietta michiganensis, while prevalences of the nematode Batracholandros magnavulvaris were 52% and<br />
30% for P. punctatus and P. wehrlei, respectively. Mean intensities were 3.4 nematodes per infected host for<br />
P. punctatus and 4.0 for P. wehrlei. Only 2 of 61 B. magnavulvaris collected were males. This is the first report<br />
of parasites from these plethodontid species and the first record of C. michiganensis from West Virginia.<br />
KEY WORDS: Batracholandros magnavulvaris, Cepedietta michiganensis, Plethodon punctatus, Plethodon<br />
wehrlei, Ciliata, Nematoda, West Virginia, U.S.A.<br />
The Cow Knob salamander, Plethodon punctatus<br />
Highton, 1972, is a large plethodontid salamander<br />
known only from higher elevations<br />
(>730 m) of the Shenandoah and Great North<br />
mountains in Augusta, Rockingham, and Shenandoah<br />
counties of Virginia (Buhlmann et al.,<br />
1988; Conant and Collins, 1991). The range of<br />
this rare species in West Virginia is restricted to<br />
higher elevations (>730 m) of Hardy and Pendleton<br />
counties in the eastern panhandle. All 25<br />
P. punctatus examined for this study were collected<br />
on Shenandoah Mountain in Pendleton<br />
County, West Virginia, from June through August<br />
1996, under a permit granted by the West<br />
Virginia Division of Natural Resources<br />
(WVDNR) and written permission from the U.S.<br />
Fish and Wildlife Service.<br />
Wehrle's salamander, Plethodon wehrlei<br />
Fowler and Dunn, 1917, is considered a near<br />
sibling of P. punctatus, and has been recorded<br />
from a wide range of elevations in 28 of West<br />
Virginia's 55 counties (Green and Pauley, 1987).<br />
The geographic range of P. wehrlei extends<br />
from southwestern New York to northwestern<br />
North Carolina (Conant and Collins, 1991). All<br />
13 P. wehrlei individuals used in this study were<br />
collected from Shaver's Mountain in Randolph<br />
County, West Virginia, from May through August<br />
1996, under a permit from the WVDNR.<br />
The original purpose of collecting these plethodontid<br />
species was to obtain reproductive and<br />
Corresponding author.<br />
185<br />
ecological data for use in forest and wildlife<br />
management plans. Because there are no published<br />
reports of parasites from either species,<br />
these collections also offered the opportunity to<br />
examine them for parasites.<br />
Materials and Methods<br />
All salamanders were anesthetized in Chloretone®<br />
within 48 hr of collection. Snout-to-vent lengths (SVL)<br />
were measured with vernier calipers to the nearest 0.1<br />
mm. Salamanders were killed by decapitation, sexed,<br />
and the small and large intestines were removed for<br />
examination. The SVL for P. punctatus (ft/mean in mm<br />
± 1 SD) was 18/63.8 ± 6.9 for males and 7/65.3 ±<br />
7.9 for females. Because the difference in mean SVL<br />
for males versus females was not significant (£0.05,23 =<br />
0.469), individuals of both host sexes were combined<br />
to calculate prevalence of ciliate infection and prevalence<br />
and mean intensity of nematode infection. The<br />
SVL for P. wehrlei (/z/mean in mm ± 1 SD) was 6/<br />
59.8 ± 9.3 for males and 7/59.9 ± 11.7 for females.<br />
Again the difference in mean SVL for males versus<br />
females was not significant (?oo5,ii = 0.017), and the<br />
data for both host sexes were combined for calculations<br />
of prevalence and mean intensity.<br />
During necropsy it was evident that both salamander<br />
species harbored astomatous ciliates and nematodes.<br />
Whole mounts of these ciliates and nematodes were<br />
prepared by staining in Semichon's acetic carmine, dehydrating<br />
in an ethanol series, clearing in xylene, and<br />
mounting in Permount®. Voucher specimens have been<br />
deposited in the U.S. National Parasite Collection,<br />
Beltsville, Maryland 20705, U.S.A., under accession<br />
numbers USNPC 89838 (Cepedietta michiganensis)<br />
and USNPC 89839 (Batracholandros magnavulvaris).<br />
Results<br />
The astomatous ciliate, C. michiganensis<br />
(Woodhead, 1928) Corliss, de Puytorac, and<br />
Copyright © 2011, The Helminthological Society of Washington
186 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Table 1. Published reports of amphibian hosts harboring Cepedietta michiganensis, with prevalences (P)<br />
by locality; host scientific names, taxonomic authorities, and dates after Frost (1985).<br />
Host common name and species<br />
Arnbystoma jeffersonianum (Green, 1 827) (Jefferson salamander)<br />
Ambystoma opacum (Gravenhorst, 1807) (marbled salamander)<br />
Desmognathus fitscus (Green, 1818) (northern dusky<br />
salamander)<br />
Desmognathus phoca (Matthes, 1855) (seal salamander)<br />
Eurycea bislineata (Green, 1818) (northern two-lined<br />
salamander)<br />
Eurycea cirrigera (Green, 1830) (southern two-lined<br />
salamander)<br />
Eurycea guttolineata (three-lined salamander):!:<br />
Hemidactylium scutatum (Temminck and Schlegel,<br />
1838) (four-toed salamander)<br />
Plethodon albagula Grobman, 1944 (western slimy salamander)<br />
Plethodon cinereus (Green, 1818) (red-backed salamander)<br />
Plethodon fourchensis Duncan and Highton, 1979<br />
(Fourche Mountain salamander)<br />
Plethodon glutinosus (Green, 1818) (northern slimy salamander)<br />
Plethodon jordani Blatchley, 1901 (Jordan's salamander)<br />
Plethodon ouachitae Dunn and Heinze, 1933 (Rich<br />
Mountain salamander)<br />
Plethodon punctatus Highton, 1972 (Cow Knob salamander)<br />
Plethodon wehrlei Fowler and Dunn, 1917 (Wehrle's<br />
salamander)<br />
Pseudotriton moiitanus Baird, 1849 (midland mud salamander)<br />
Rana clamitans Latreille, 1801 (southern green frog)<br />
Rana sylvatica LeConte, 1825 (wood frog)<br />
P(%)<br />
1/NI*<br />
7<br />
0/12|<br />
6<br />
9<br />
6<br />
18<br />
16<br />
0/2 It<br />
70<br />
1/NI*<br />
9<br />
18<br />
33<br />
5<br />
1 l/7t<br />
41-72§<br />
20<br />
14<br />
0-49<br />
48<br />
60<br />
23<br />
33<br />
—<br />
20<br />
Michigan<br />
North Carolina<br />
North Carolina<br />
Locality<br />
North Carolina<br />
North Carolina<br />
New Hampshire<br />
North Carolina<br />
North Carolina<br />
North Carolina<br />
Michigan<br />
Reference<br />
Woodhead, 1928<br />
Rankin, 1937a<br />
Rankin, 1937a<br />
Rankin, 1937a<br />
Rankin, 1937a<br />
Muzzall et al., 1997<br />
Mann, 1932<br />
Mann, 1932<br />
Rankin, 1937a<br />
Woodhead, 1928<br />
Arkansas McAllister et al., 1993<br />
Ohio<br />
New Hampshire<br />
Michigan<br />
Arkansas<br />
North Carolina<br />
North Carolina<br />
Tennessee<br />
Arkansas<br />
Great Smoky Mts. of Tennessee-<br />
North Carolina<br />
Tennessee<br />
Arkansas<br />
West Virginia<br />
West Virginia<br />
North Carolina<br />
Ohio<br />
Ohio<br />
Hazard, 1937<br />
Muzzall et al., 1997<br />
Muzzall, 1990<br />
Winter et al., 1986<br />
Mann, 1932<br />
Rankin, 1937a<br />
Powders, 1970<br />
Winter et al., 1986<br />
Powders, 1967<br />
Powders, 1970<br />
Winter et al., 1986<br />
Present study<br />
Present study<br />
Rankin, 1937a<br />
Odlaug, 1954<br />
Hazard, 1937<br />
* Single individual infected/sample size not included.<br />
t Prevalence in Durham, North Carolina, U.S.A. area of the central Piedmont/prevalence in the mountains.<br />
:!: Eurycea guttolineata, the three-lined salamander, is considered a subspecies of the long-tailed salamander, E. longicauda<br />
(Green, 1818).<br />
§ Prevalences inversely related to altitude and higher in fall months.<br />
Lorn, 1965, was found in 60% (15/25) and 23%<br />
(3/13) of P. punctatus and P. wehrlei, respectively<br />
(Table 1). This ciliate species was primarily<br />
aggregated in the duodenum of both host<br />
species, either free in the lumen or attached to<br />
the intestinal epithelium. Cepedietta michiganensis<br />
individuals were often so numerous that<br />
they appeared to occlude the duodenum; however,<br />
because there was no gross distention of<br />
this organ it was unlikely that any blockage of<br />
Copyright © 2011, The Helminthological Society of Washington<br />
food materials actually occurred. No damage to<br />
cells of the host's intestinal epithelium was evident.<br />
There were a few instances where small<br />
numbers of ciliates were found in the middle and<br />
posterior small intestine, large intestine, or gall<br />
bladder of P. punctatus, as well.<br />
The oxyurid nematode, Batracholandros<br />
magnavulvaris (Schad, 1960) Fetter and Quentin,<br />
1976, was found in the large intestine of<br />
52% (13/25) P. punctatus and 31% (4/13) P.
JOY AND TUCKER—SALAMANDER PARASITES 187<br />
Table 2. Published reports of hosts harboring Batracholandros magnavulvaris, with prevalences (P) and<br />
mean intensities (x) by locality; host scientific names, taxonomic authorities, and dates after Frost (1985).<br />
Host common name and species<br />
Aneides flavipunctatus (Strauch, 1870) (black salamander)<br />
Aneides aeneus Cope and Packard, 1881 (green salamander)<br />
Desmognathus brimleyorum Stejneger, 1895 (Ouachita<br />
dusky salamander)<br />
Desmognathus fuscus (Green, 1818) (northern dusky salamander)<br />
Desmognathus monticola Dunn, 1916 (seal salamander)<br />
Desmognathus ochrophaeus Cope, 1959 (Allegheny<br />
Mountain dusky salamander)<br />
Desmognathus phoca (Matthes, 1855) (seal salamander)<br />
Desmognathus quadrimaculatus Holbrook, 1840 (blackbellied<br />
salamander)<br />
Eurycea bislineata (Green, 1818) (northern two-lined<br />
salamander)<br />
Eurycea guttolineata (three-lined salamander)<br />
Eurycea lucifuga Rafinesque, 1822 (cave salamander)<br />
Leiirognathus marmoratus Moore, 1 899 (shovel-nosed<br />
salamander)<br />
Notophthalmus viridescens (Rafinesque, 1820) (red-spotted<br />
newt, red eft)<br />
Notophthalmus viridescens (red-spotted newt, adult)<br />
Plethodon caddoensis Pope and Pope, 1951 (Caddo<br />
Mountain salamander)<br />
Plethodon cinereus (Green, 1818) (red-backed salamander)<br />
Plethodon fourchensis Duncan and Highton, 1 979<br />
(Fourche Mountain salamander)<br />
Plethodon glutinosus (Green, 1818) (northern slimy salamander)<br />
Plethodon ouachitae Dunn and Heinze, 1933 (Rich<br />
Mountain salamander)<br />
Plethodon punctatus Highton, 1 972 (Cow Knob salamander)<br />
Plethodon serratus Grobman, 1944 (southern red-backed<br />
salamander)<br />
Plethodon wehrlei Fowler and Dunn, 1917 (Wehrle's salamander)<br />
Plethodon yonahlossee Dunn, 1917 ( Yonahlossee salamander)<br />
P (%)<br />
50<br />
24<br />
77<br />
30<br />
48<br />
—<br />
10<br />
6<br />
27<br />
—<br />
48<br />
60<br />
50<br />
23<br />
14<br />
40<br />
14<br />
25<br />
15<br />
7<br />
31<br />
27<br />
11<br />
2<br />
0/64±<br />
100<br />
6<br />
100<br />
8<br />
12<br />
0/2*<br />
50<br />
28<br />
—<br />
9<br />
33<br />
0/3$<br />
14<br />
52<br />
22<br />
31<br />
33<br />
A'<br />
3<br />
5<br />
3<br />
188 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
wehrlei individuals (Table 2). Mean intensities<br />
(±1 SD) were 3.4 (±1.9) and 4.0 (±4.0) for P.<br />
punctatus and P. wehrlei, respectively. Of the 61<br />
B. magnavulvaris individuals collected, only 2<br />
(both in P. wehrlei) were males.<br />
Discussion<br />
Prevalences of 60% and 23% for C. michiganensis<br />
in P. punctatus and P. wehrlei from<br />
West Virginia fall within ranges cited by previous<br />
investigators for this species (Table 1). Finding<br />
C. michiganensis species throughout the intestinal<br />
tract, with heavy aggregations in the duodenum,<br />
is consistent with the observations of<br />
Winter et al. (1986), who noted that C. michiganensis<br />
could be found throughout the intestine<br />
of Plethodon ouachitae Dunn and Heinze, 1933,<br />
but was concentrated in the anterior third of this<br />
organ. There was a single case of C. michiganensis<br />
infection in the gallbladder of P. punctatus,<br />
but ciliates were not attached to the epithelium<br />
of the gall bladder. Winter et al. (1986)<br />
also reported C. michiganensis from the gall<br />
bladder of P. ouachitae, adding that the ciliates<br />
were not attached to the epithelium. Three other<br />
reports mentioned occurrence of this ciliate in<br />
the host's gall bladder: Muzzall (1990) for Plethodon<br />
cinereus (Green, 1818); McAllister et al.<br />
(1993) for Plethodon albagula Grobman, 1944;<br />
and Muzzall et al. (1997) for Eurycea bislineata<br />
(Green, 1818) and P. cinereus. Previous reports<br />
of amphibian hosts harboring species of C. michiganensis<br />
are summarized in Table 1.<br />
Prevalences of 52% and 31% recorded in the<br />
present study for B. magnavulvaris in P. punctatus<br />
and P. wehrlei, respectively, are not unusual.<br />
This nematode species exhibits little host<br />
specificity and is found in widely varying prevalences<br />
(McAllister et al., 1995), an observation<br />
supported by the findings of other investigators<br />
summarized in Table 2. McAllister et al. (1995)<br />
also reported that prevalence for B. magnavulvaris<br />
in Desmognathus brimleyorum Stejneger,<br />
1895 varies seasonally, being highest (34%) in<br />
mid-March versus only 17% for late May. While<br />
we had relatively few salamanders in our sample,<br />
all B. magnavulvaris individuals were collected<br />
from 13 of 21 P. punctatus in June and<br />
July. Similarly, 4 of the 9 P. wehrlei examined<br />
in May through July were infected. None of the<br />
8 plethodontids (4 P. punctatus and 4 P. wehrlei)<br />
sampled in August were infected, suggesting<br />
that the variations in prevalence by season noted<br />
Copyright © 2011, The Helminthological Society of Washington<br />
by McAllister et al. (1995) may be a normal<br />
pattern. Mean intensities of infection at 3.4 and<br />
4.0 for P. punctatus and P. wehrlei, respectively,<br />
were relatively high compared with previous reports<br />
(Table 2). Only 2 B. magnavulvaris individuals<br />
collected in the present study were<br />
males. This heavily female-biased sex ratio for<br />
B. magnavulvaris is similar to the observations<br />
of previous investigators (Dyer et al., 1980;<br />
Muzzall, 1990; Joy et al., 1993; McAllister et<br />
al., 1995).<br />
Acknowledgments<br />
We are grateful to William Tolin, U.S. Fish<br />
and Wildlife Service, for granting the requisite<br />
collection permit for P. punctatus, and to Thomas<br />
Pauley for reviewing this manuscript.<br />
Literature Cited<br />
Buhlmann, K. A., C. A. Pague, J. C. Mitchell, and<br />
R. B. Glascow. 1988. Forestry operations and terrestrial<br />
salamanders: techniques in a study of the<br />
Cow Knob salamander, Plethodon punctatus. Pages<br />
38-44 in R. C. Szaro, K. E. Severson, and D.<br />
R. Patton, eds. Management of Amphibians, Reptiles<br />
and Mammals in North America. USDA Forest<br />
Service, Rocky Mountain Forest and Range<br />
Experiment Station, Fort Collins, Colorado,<br />
U.S.A. Technical Report RM-166.<br />
Bursey, C. R., and D. R. Schibli. 1995. A comparison<br />
of the helminth fauna of two Plethodon cinereus<br />
populations. Journal of the Helminthological Society<br />
of Washington 62:232-236.<br />
Conant, R., and J. T. Collins. 1991. A Field Guide<br />
to Reptiles and Amphibians of Eastern and Central<br />
North America. Houghton Mifflin Company,<br />
Boston, Massachusetts, U.S.A. 450 pp.<br />
Dunbar, J. R., and J. D. Moore. 1979. Correlations<br />
of host specificity with host habitat in helminths<br />
parasitizing the plethodontids of Washington<br />
County, Tennessee. Journal of the Tennessee<br />
Academy of Science 54:106-109.<br />
Dyer, W. G. 1991. Helminth parasites of amphibians<br />
from Illinois and adjacent midwestern states.<br />
Transactions of the Illinois <strong>State</strong> Academy of Science<br />
84:1-19.<br />
, R. A. Brandon, and R. L. Price. 1980. Gas<br />
trointestinal helminths in relation to sex and age<br />
of Desmognathus fuxcus (Green, 1818) from Illinois.<br />
Proceedings of the Helminthological Society<br />
of Washington 47:95-99.<br />
-, and S. B. Peck. 1975. Gastrointestinal parasites<br />
of the cave salamander, Eurycea lucifuga Rafinesque,<br />
from the southeastern United <strong>State</strong>s. Canadian<br />
Journal of Zoology 53:52-54.<br />
Ernst, E. M. 1974. The parasites of the red-backed<br />
salamander, Plethodon cinereus. Bulletin of the<br />
Maryland Herpetological Society 10:108-1 14.<br />
Fischthal, J. H. 1955. Ecology of worm parasites in
south-central New York salamanders. American<br />
Midland Naturalist 53:176-183.<br />
Frost, D. R. 1985. Amphibian Species of the World:<br />
A Taxonomic and Geographic Reference. Allen<br />
Press, Inc. and The Association of Systematics<br />
Collections, Lawrence, Kansas, U.S.A. 732 pp.<br />
Goater, T. M., G. W. Esch, and A. O. Bush. 1987.<br />
Helminth parasites of sympatric salamanders: ecological<br />
concepts at infracommunity, component<br />
and compound community levels. American Midland<br />
Naturalist 118:289-300.<br />
Green, N. B., and T. K. Pauley. 1987. Amphibians<br />
and Reptiles in West Virginia. University of Pittsburgh<br />
Press, Pittsburgh, Pennsylvania, U.S.A. 241<br />
pp.<br />
Hazard, F. O. 1937. Two new host records for the<br />
protozoan Haptophyra michiganensis Woodhead.<br />
Journal of <strong>Parasitology</strong> 23:315-316.<br />
Joy, J. E., T. K. Pauley, and M. L. Little. 1993.<br />
Prevalence and intensity of Thelandros magncivulvaris<br />
and Omeia papillocaucia (Nematoda) in<br />
two species of desmognathine salamanders from<br />
West Virginia. Journal of the Helminthological<br />
Society of Washington 60:93-95.<br />
Lehmann, D. L. 1954. Some helminths of West Coast<br />
urodeles. Journal of <strong>Parasitology</strong> 40:231.<br />
Mann, D. R. 1932. The ecology of some North Carolina<br />
salamanders with special reference to their<br />
parasites. M.A. Thesis, Duke University, Durham,<br />
North Carolina, U.S.A. 52 pp.<br />
McAllister, C. T., C. R. Bursey, S. J. Upton, S. E.<br />
Trauth, and D. B. Conn. 1995. Parasites of' Desmognathus<br />
briinleyorum (Caudata: Plethodontidae)<br />
from the Ouachita Mountains of Arkansas<br />
and Oklahoma. Journal of the Helminthological<br />
Society of Washington 62:150-156.<br />
, S. J. Upton, and S. E. Trauth. 1993. Endoparasites<br />
of western slimy salamanders, Plethodon<br />
albagula (Caudata: Plethodontidae), from<br />
JOY AND TUCKER—SALAMANDER PARASITES 189<br />
Arkansas. Journal of the Helminthological Society<br />
of Washington 60:124-126.<br />
Muzzall, P. M. 1990. Endoparasites of the red-backed<br />
salamander, Plethodon c. cinereus, from southwestern<br />
Michigan. Journal of the Helminthological<br />
Society of Washington 57:165-167.<br />
, C. R. Peebles, and T. M. Burton. 1997. Endoparasites<br />
of plethodontid salamanders from Paradise<br />
Brook, New Hampshire. Journal of <strong>Parasitology</strong><br />
83:1193-1195.<br />
Odlaug, T. O. 1954. Parasites of some Ohio Amphibia.<br />
Ohio Journal of Science 54:126-128.<br />
Powders, V. N. 1967. Altitudinal distribution of the<br />
astomatous ciliate Cepedietta michiganensis<br />
(Woodhead) in a new host, Plethodon jordani<br />
Blatchley. Transactions of the American Microscopical<br />
Society 86:336—338.<br />
. 1970. Altitudinal distribution of the protozoan<br />
Cepedietta michiganensis in the salamanders<br />
Plethodon glutinosus and Plethodon jordani in<br />
eastern Tennessee. American Midland Naturalist<br />
83:393-403.<br />
Rankin, J. S., Jr. 1937a. An ecological study of parasites<br />
of some North Carolina salamanders. Ecological<br />
Monographs 7:169-270.<br />
. 1937b. New helminths from North Carolina<br />
salamanders. Journal of <strong>Parasitology</strong> 23:29-42.<br />
Schad, G. A. 1963. Thelandros magnavidvaris (Rankin,<br />
1937) Schad, 1960 (Nematoda: Oxyuroidea)<br />
from the green salamander, Aneides aeneus. Canadian<br />
Journal of Zoology 41:943-946.<br />
Walton, A. C. 1940. Some nematodes from Tennessee<br />
Amphibia. Journal of the Tennessee Academy of<br />
Science 15:402-405.<br />
Winter, E. A., W. M. Zawada, and A. A. Johnson.<br />
1986. Comparison of the symbiotic fauna of the<br />
family Plethodontidae in the Ouachita Mountains<br />
of western Arkansas. Proceedings of the Arkansas<br />
Academy of Sciences 40:82-85.<br />
Woodhead, A. F. 1928. Haptophyra michiganensis sp.<br />
nov., a protozoan parasite of the four-toed salamander.<br />
Journal of <strong>Parasitology</strong> 14:177-182.<br />
Copyright © 2011, The Helminthological Society of Washington
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 190-195<br />
Some Adult Endohelminths Parasitizing Freshwater Fishes from the<br />
Atlantic Drainages of Nicaragua<br />
M. LEOPOLDINA AGUIRRE-MACEDO, M TOMAS ScHOLZ,1-2 DAVID GONZALEZ-SOUS,'<br />
VICTOR M. VIDAL-MARTINEZ,' PETR PosEL,3 GREGORY ARJONA-TORRES,'<br />
SVETLANA DUMAILO,3 AND EDUARDO SlU-ESTRADA3<br />
1 Laboratorio de Parasitologfa, Centre de Investigacion y Estudios Avanzados del Institute Politecnico<br />
Nacional (CINVESTAV-IPN), Unidad Merida, Carretera Antigua a Progreso Km 6, A.P. 73 Cordemex, C.P.<br />
97310, Merida, Yucatan, Mexico (e-mail: leo@mda.cinvestav.mx),<br />
2 Institute of <strong>Parasitology</strong>, Academy of Sciences of the Czech Republic, Branisovska 31, 370 05 Ceske<br />
Budejovice, Czech Republic (e-mail: tscholz@paru.cas.cz), and<br />
3 Bluefields Indian and Caribbean University, P.O. Box 88, Bluefields, Nicaragua (e-mail: bicu@ibw.com.ni)<br />
ABSTRACT: Adults of 12 endoparasitic helminths were recorded from 8 freshwater fish species from the South<br />
Atlantic Autonomous Region, Nicaragua: 8 digeneans Crassicutis cichlasomae, Magnivitellinum simplex, Oligogonotylus<br />
manteri, Prosthenhystera obesa, Saccocoelioides sogandaresi, Saccocoelioides sp. 1, Saccocoelioides<br />
sp. 2, and Allocreadiidae gen. sp. ("Crepidostomum" sp.); 3 nematodes Procamallanus (Spirocamallanus)<br />
rebecae, Procamallanus (Spirocamallanus) neocaballeroi, and Rhabdochona kidderi kidderi; and 1 acanthocephalan<br />
Neoechinorhynchus golvani. <strong>Comparative</strong> measurements among 5. sogandaresi from Poecilia velifera,<br />
Saccocoelioides sp. 1 from Cichlasoma maculicaiida, and Saccocoelioides sp. 2 from Astyanax fasciatus, as well<br />
as drawings of the 2 latter species are given for future reference. All but C. cichlasomae and O. manteri are<br />
reported from Nicaragua for the first time, and most taxa also represent new geographical records for Central<br />
America. The majority of species have previously been found in freshwater fishes from southeastern Mexico,<br />
which indicates a close similarity of the helminth faunas of both regions, in accordance with previous data on<br />
the larval stages of endohelminths and gill monogeneans.<br />
KEY WORDS: Helminths, parasites, Digenea, Nematoda, Acanthocephala, freshwater fishes, Nicaragua.<br />
During a short visit by 4 of the authors<br />
(M.L.A.M., T.S., V.M.V.M., and G.A.T.) to Nicaragua<br />
in March 1999, brackish and freshwater<br />
fishes from the Autonomous Region of the<br />
South Atlantic were examined for helminth parasites.<br />
Because information on helminths parasitizing<br />
freshwater fishes in Nicaragua is limited<br />
to the report by Watson (1976), in which several<br />
species of trematodes from Lake Nicaragua were<br />
reported, a list of adult endohelminths is provided,<br />
with morphological data on some taxa. The<br />
results of a survey of larval stages of endohelminths<br />
found in the same fish hosts and ancyrocephaline<br />
monogeneans from the gills of cichlids,<br />
have already been published (Aguirre-Macedo<br />
et al., <strong>2001</strong>; Vidal-Martinez, Scholz, and<br />
Aguirre-Macedo, <strong>2001</strong>).<br />
Materials and Methods<br />
A total of 56 fish of the following 8 species was<br />
examined: tetra Astyanax fasciatus (Cuvier, 1819) (8<br />
specimens examined) (family Characidae); pastel cichlid<br />
Amphilophus alfari (Meek, 1907) (3); convict cichlid<br />
4 Corresponding author.<br />
190<br />
Copyright © 2011, The Helminthological Society of Washington<br />
Archocentrus nigrofasciatus (Giinther, 1869) (3); blackbelt<br />
cichlid Cichlasoma maculicaiida (Regan, 1805)<br />
(12); jaguar cichlid Cichlasoma managuense (Giinther,<br />
1867) (13); butterfly cichlid Herotilapia multispinosa<br />
(Giinther, 1867) (8) (Cichlidae); molly Poecilia velifera<br />
(Regan, 1814) (5) (Poeciliidae); and long-whiskered catfish<br />
Rhamdia nicaraguensis (Giinther, 1864) (4) (Pimelodidae).<br />
Fish were collected by hook and line and<br />
throw nets from 7 localities of the Atlantic drainages of<br />
Nicaragua in the Autonomous Region of the South Atlantic<br />
(Region Autonoma del Atlantico Sur—RAAS):<br />
Torsuani River (11°47'06"N; 83°52'38"W); Mahogany<br />
River (12°03'22"N; 83°59'07"W); Cano Negro Stream<br />
(12°00'55"N; 84°01'10"W), in Bluefields City; Walpatara<br />
Bridge (12°00'14"N; 83°45'58"W); Loonku Creek<br />
(11°59'05"N; 83°46'48"W); Cano Marandn Stream<br />
(12°00'10"N; 83°46'39"W); and Puente Chino<br />
(12°00'30"N; 83°46'13"W). The number offish sampled<br />
in individual localities and map locations are given in<br />
Aguirre-Macedo et al. (<strong>2001</strong>).<br />
Fish were transported alive to the laboratory of the<br />
Bluefields Indian and Caribbean University (BICU),<br />
where they were examined by routine helminthological<br />
procedures outlined by Vidal-Martinez, Aguirre-Macedo<br />
et al. (<strong>2001</strong>). All helminths were studied in fresh<br />
preparations and counted in situ. Adult helminths were<br />
considered those with fully developed reproductive organs<br />
regardless of the presence of eggs. Eventually,<br />
some digeneans were fixed with a glycerin-ammonium<br />
picrate (GAP) mixture following the methodology out-
AGUIRRE-MACEDO ET AL.—ENDOHELMINTHS FROM NICARAGUAN FISHES 191<br />
Table 1. Some endoparasitic helminths collected from Nicaraguan freshwater fishes.<br />
Digcnea<br />
Helminth species<br />
Crassicutis cichlasomae Manter, 1936<br />
Magnivitellinum simplex Kloss, 1966<br />
Oligogonotylus manteri Watson, 1976<br />
IProsthenhystera obesa (Diesing, 1850)<br />
Saccocoelioides sogandaresi Lumsden,<br />
1964<br />
Saccocoelioides sp. 1<br />
Saccocoelioides sp. 2<br />
Allocreadiidae gen. sp.<br />
Nematoda<br />
Procamallanus (Spirocamallanus) rebecae<br />
Andrade-Salas, Pineda-Lopez, and<br />
Osorio-Sarabia, 1994<br />
Procamallanus (Spirocamallanus) neocaballeroi<br />
(Caballero-Deloya, 1977)<br />
Rhabdochona kidderi kidderi Pearsc,<br />
1936<br />
Acanthocephala<br />
Neoechinorhynchus golvani Salgado-Maldonado,<br />
1978<br />
Hosts<br />
(no. infected/examined)<br />
Cichlasoma maculicauda (5/7)<br />
Astyanax fasciatus ( 1 /4)<br />
(1/2)<br />
C. maculicauda (2/7)<br />
Cichlasoma managuense (1/2)<br />
A. fasciatus (1/2)<br />
Poet-ilia velifera (1/1)<br />
C. maculicauda (1/3)<br />
(6/7)<br />
A. fasciatus (2-4)<br />
A. fasciatus ( 1 /4)<br />
(1/2)<br />
Intensity<br />
range<br />
1-8<br />
1<br />
6<br />
3-6<br />
1<br />
1<br />
1<br />
17<br />
4-42<br />
1-3<br />
1<br />
4<br />
Amphilophus alfari (1/3) 29<br />
C. maculicauda (21 \) 1—2<br />
Herotilapia rnultispinosa (3/8) 1—3<br />
A. fasciatus (1/9) 3<br />
C. maculicauda (1/11) 1<br />
C. maculicauda (1/11) 1<br />
A. alfari (1/1)<br />
C. managuense (1/2)<br />
C. managuense (2/4)<br />
H. rnultispinosa (1/4)<br />
H. rnultispinosa (1/3)<br />
lined by Ergens (1969). Measurements are given in<br />
micrometers. Drawings were made with the aid of a<br />
drawing tube. Reference specimens were deposited in<br />
the Coleccion Nacional de Helmintos (CNHE), Mexico<br />
City, Mexico, and the Laboratory of <strong>Parasitology</strong>,<br />
CINVESTAV-IPN (CHCM), Merida, Mexico.<br />
Results<br />
A total of 12 helminth species was found.<br />
Data on the hosts, localities, and infection range<br />
are provided in Table 1. All but 1 species<br />
(IProsthenhystera obesa) were located in the intestine;<br />
IP. obesa inhabited the gall blader.<br />
Among the species found, 3 trematodes were<br />
not identified to species level: 2 species of Saccocoelioides<br />
and a trematode of the subfamily<br />
Allocreadiinae (Allocreadiidae). Measurements<br />
of the first 2 species (Figs. 1-4), together with<br />
those of a congeneric species {Saccocoelioides<br />
sogandaresi) from Poecilia velifera are presented<br />
in Table 2.<br />
1<br />
1<br />
1-8<br />
3<br />
2<br />
Locality<br />
Cano Maranon<br />
Torsuani River<br />
Loonku Creek<br />
Torsuani River<br />
Puente Chino<br />
Loonku Creek<br />
Cano Maranon<br />
Torsuani River<br />
Cano Maranon<br />
Torsuani River<br />
Torsuani River<br />
Loonku Creek<br />
Torsuani River<br />
Loonku Creek<br />
Deposit<br />
accession nos.<br />
CNHE/CHCM<br />
4193/<br />
4 1 95/<br />
4196/<br />
4194/<br />
/383<br />
/384<br />
/380<br />
4142/<br />
/393, 393-1<br />
Mahogany River 4143/<br />
Torsuani River /396<br />
Torsuani River<br />
Loonku Creek<br />
Puente Chino<br />
Cano Negro<br />
Loonku Creek<br />
Puente Chino<br />
41911<br />
Discussion<br />
The number of species of adult endohelminths<br />
recorded was lower than that of larval stages, in<br />
particular metacercariae of digeneans, found in<br />
the same fish hosts (Aguirre-Macedo et al.,<br />
<strong>2001</strong>). Adult trematodes represented the dominant<br />
group (8 species) in this study, whereas<br />
nematodes were fewer (only 3 species). Only 1<br />
acanthocephalan occurred in fishes examined.<br />
No adult cestodes were found, even in the pimelodid<br />
catfish Rhamdia nicaraguensis, but<br />
only 4 fish of this species were examined. Thus,<br />
it is probable that more fish and localities need<br />
to be sampled to find adult cestodes, especially<br />
considering the low prevalence (
192 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Copyright © 2011, The Helminthological Society of Washington
AGUIRRE-MACEDO ET AL.—ENDOHELMINTHS FROM NICARAGUAN FISHES 193<br />
Table 2. Measurements of species of Saccocoelioides from Nicaraguan freshwater fishes (n = number of<br />
specimens measured).<br />
Host<br />
Body shape<br />
Body length<br />
Maximum width<br />
Oral sucker<br />
Ventral sucker<br />
Sucker ratio<br />
Position of acctabulum<br />
Prepharynx<br />
Pharynx<br />
Oral sucker/pharynx ratio<br />
Extent of ceca<br />
Tcstis<br />
Hermaphroditic sac<br />
Ovary<br />
Extent of vitellarium<br />
Eggs<br />
Saccocoelioides<br />
sogandaresi<br />
(n = 1)<br />
Poecilia velifera*<br />
Widely oval<br />
995<br />
310<br />
100 X 122<br />
123 X 130<br />
1.14:1<br />
48% of body length<br />
38<br />
90 X 98<br />
1.18:1<br />
Anterior line of testis<br />
102 X 82<br />
150 X 76<br />
—<br />
—<br />
—<br />
Specimens fixed with GAP under pressure.<br />
Ponce de Leon, 1996) from fish of the genus<br />
Rhamdia in the Yucatan Peninsula (see Scholz<br />
et al., 1996).<br />
In South America, cestodes appear to be the<br />
dominant component of the fauna of endohelminths<br />
in freshwater fishes, in terms of the number<br />
of species and genera (Thatcher, 1991; Rego<br />
et al., 1999). These cestodes belong almost exclusively<br />
to the order Proteocephalidea, and they<br />
occur most frequently in siluriform fishes, including<br />
pimelodids (de Chambrier and Vaucher,<br />
1999; Rego, 2000).<br />
The endohelminth fauna of fishes from the<br />
Atlantic coastal drainages of Nicaragua closely<br />
resembles that of southeastern Mexico. Similar<br />
to the larval stages of endohelminths (Aguirre-<br />
Macedo et al., <strong>2001</strong>), a majority of species<br />
found occur in congeneric fish hosts from the<br />
Yucatan Peninsula (Moravec et al., 1995; Scholz<br />
et al., 1995, 1996; Salgado-Maldonado et al.,<br />
1997; Scholz and Vargas-Vazquez, 1998; Vidal-<br />
Martinez, Aguirre-Macedo et al., <strong>2001</strong>). This<br />
similarity indicates close relationships between<br />
Saccocoelioides sp. 1<br />
(n = 3)<br />
Astyanax fasciatus<br />
Elongate, with tapering ends<br />
1,070-1,210<br />
290-320<br />
90-112 X 100-120<br />
105-115 X 113-125<br />
0.83-1.01:1<br />
33-35%<br />
45-50<br />
70-78 X 63-69<br />
1.35-1.61:1<br />
About midline of testis<br />
274-290 X 140-173<br />
245-280 X 143-160<br />
98-105 X 80-104<br />
Far postacetabular<br />
73-75 X 46-50<br />
Saccocoelioides sp. 2<br />
(« = 15)<br />
Cichlasoma maculicauda<br />
Oval to elongate<br />
470-<strong>68</strong>0<br />
150-335<br />
67-105 X 75-125<br />
56-120 X 50-125<br />
0.63-1.30:1<br />
31-39%<br />
36-72<br />
50-82 X 47-75<br />
1.01-1.46:1<br />
About midline to % of testis<br />
72-175 X 58-145<br />
77-162 X 62-135<br />
37-87 X 35-87<br />
About midline of acetabulum<br />
67-81 X 36-47<br />
the helminth faunas of freshwater fishes in Central<br />
America and southeastern Mexico, in accordance<br />
with the analysis of Vidal-Martinez and<br />
Kennedy (2000) and the general biogeography<br />
of the neotropics (Briggs, 1984). Vidal-Martinez,<br />
Scholz and Aguirre-Macedo (<strong>2001</strong>) also<br />
found a marked resemblance between gill monogeneans<br />
of cichlids from Nicaragua and those<br />
from Yucatan.<br />
Three species of trematodes, Magnivitellinum<br />
simplex, IP. obesa, and S. sogandaresi, all nematodes,<br />
and the acanthocephalan Neoechinorhynchus<br />
golvani, previously found in North and<br />
South America (Travassos et al., 1969; Thatcher,<br />
1991; Perez-Ponce de Leon et al., 1996; Salgado-Maldonado<br />
et al., 1997; Moravec, 1998), are<br />
reported from Central America for the first time.<br />
With the exception of the trematodes Oligogonotylus<br />
manteri and Crassicutis cichlasomae reported<br />
from Lake Nicaragua by Watson (1976),<br />
all species also represent new geographical records<br />
from Nicaragua. This reflects the shortage<br />
Figures 1—4. 1, 2. Saccocoelioides sp. 1 from Cichlasoma maculicauda. 1. Total view from the ventral<br />
aspect. 2. Tegumental spines at pharyngeal level. 3, 4. Saccocoelioides sp. 2 from Astyanax fasciatus. 3.<br />
Total view ventral from the ventral aspect. 4. Tegumental spines at pharyngeal level. Scale bars in millimeters.<br />
Copyright © 2011, The Helminthological Society of Washington
194 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
of data on helminths of freshwater fishes in this<br />
country and in Central America in general.<br />
Three species of the genus Saccocoelioid.es<br />
Szidat, 1954, were found in this study, differing<br />
from each other in the size and shape of the<br />
body, their spination, relative size of the pharynx<br />
and hermaphroditic sac, extent of the vitellaria,<br />
egg size, etc. However, 2 of them (Figs.<br />
1-4) remain unidentified to species level because<br />
they differ from all hitherto described taxa<br />
(see Szidat, 1954; Lunaschi, 1984). They are not<br />
described as new species because of the unsatisfactorily<br />
resolved taxonomy of the genus, with<br />
numerous taxa having been inadequately described.<br />
To provide data for subsequent species<br />
identification, measurements of all 3 species are<br />
provided in Table 2.<br />
The allocreadiid trematode found in Astyanax<br />
fasciatus most resembles in its morphology the<br />
species Crepidostomum platense Szidat, 1954,<br />
and Crepidostomum stenopteri Mane-Garzon<br />
and Gascon, 1973, described from the intestine<br />
of several pimelodid catfishes from Argentina<br />
and from the characid fish "dentudo transparente"<br />
Charax (=Asiphonichthys) stenopterus<br />
(Cope, 1894) from Uruguay, respectively (see<br />
Szidat, 1954; Mane-Garzon and Gascon, 1973).<br />
However, there are marked differences among<br />
the present specimens and both species of Crepidostomum<br />
in the extent of the vitelline follicles,<br />
position of the ventral sucker, and in the<br />
shape and relative positions of the testes. It is,<br />
therefore, probable that the trematode from Nicaragua<br />
represents at least a new species. Nevertheless,<br />
it is not described formally in this paper<br />
because it differs, as do both species from<br />
South America, in its morphology from members<br />
of Holarctic species of the genus Crepidostomum<br />
Braun, 1900, and may even represent a<br />
different genus.<br />
The present report is the second on adults of<br />
helminth parasites from Nicaragua, following<br />
that of Watson (1976). It is evident that much<br />
more data on the fish parasites from larger samples<br />
of fish hosts must be obtained for better<br />
understanding of their species composition and<br />
relationships to those from other areas of the<br />
Neotropical region.<br />
Acknowledgments<br />
The authors are indebted to Dr. Anindo<br />
Choudhury, National Wildlife Health Center,<br />
Madison, Wisconsin, U.S.A., for valuable advice<br />
Copyright © 2011, The Helminthological Society of Washington<br />
and helpful suggestions. Thanks are also due to<br />
the students of the School of Marine Biology of<br />
BICU for help in collecting fish, and to the authorities<br />
of this university for enabling us to use<br />
its facilities. A short visit by M.L.A.M. and<br />
V.M.V.M. to the Czech Republic in August 1999<br />
was supported by the Institute Mexicano de<br />
Cooperacion Internacional de la Secretaria de<br />
Relaciones Exteriores, Mexico.<br />
Literature Cited<br />
Aguirre-Macedo, M. L., T. Scholz, D. Gonzalez-Solis,<br />
V. M. Vidal-Martinez, P. Posel, G. Arjona-<br />
Torres, E. Siu-Estrada, and S. Dumailo. <strong>2001</strong>.<br />
Larval helminths parasitizing freshwater fishes<br />
from the Atlantic coast of Nicaragua. <strong>Comparative</strong><br />
<strong>Parasitology</strong> <strong>68</strong>:42-51.<br />
Briggs, J. C. 1984. Freshwater fishes and biogeography<br />
of Central America and the Antilles. Systematic<br />
Zoology 33:428-435.<br />
de Chambrier, A., and C. Vaucher. 1999. Proteocephalidae<br />
et Monticelliidae (Eucestoda: Proteocephalidea)<br />
parasites des poissons d'eau douce au<br />
Paraguay, avec descriptions d'un genre nouveau<br />
et de dix especes nouvelles. Revue Suisse de<br />
Zoologie 106:165-240.<br />
Ergens, R. 1969. The suitability of ammonium-picrate<br />
in preparing slides of lower Monogenoidea. Folia<br />
Parasitologica 16:320.<br />
Lunaschi, L. I. 1984. Helmintos parasitos de peces de<br />
agua dulce de la Argentina I. Tres nuevas especies<br />
del genero Saccocoelioides Szidat, 1954 (Trematoda<br />
Haploporidae). Neotropica 30(83):31-42.<br />
Mane-Garzon, F., and A. Gascon. 1973. Digenea de<br />
peces de agua dulce del Uruguay, I. Una nueva<br />
especie del genero Crepidostomum Braum, 1900<br />
del intestine de Asiphonichthys stenopterus. Revista<br />
de Biologia del Uruguay 1:11-14.<br />
Moravec, F. 1998. Nematodes of Freshwater Fishes<br />
of the Neotropical Region. Academia, Prague,<br />
Czech Republic. 464 pp.<br />
, C. Vivas-Rodriguez, T. Scholz, J. Vargas-<br />
Vazquez, E. Mendoza-Franco, and D. Gonzalez-SoIis.<br />
1995. Nematodes parasitic in fishes of<br />
cenotes (=sinkholes) of the Peninsula of Yucatan,<br />
Mexico. Part 1. Adults. Folia Parasitologica 42:<br />
115-129.<br />
Perez-Ponce de Leon, G., L. Garcia-Prieto, D. Osorio-Sarabia,<br />
and V. Leon-Regagnon. 1996. Listados<br />
Faunisticos de Mexico. VI. Helmintos Parasitos<br />
de Peces de Aguas Continentales de Mexico.<br />
Institute de Biologia, Universidad Nacional<br />
Autonoma de Mexico, Mexico, D.F., Mexico. 100<br />
pp.<br />
Rego, A. A. 2000. Cestode parasites of neotropical<br />
teleost freshwater fishes. Pages 135-154 in A. N.<br />
Garcfa-Aldrete, G. Salgado-Maldonado, and V. M.<br />
Vidal-Martmez, eds. Metazoan Parasites in the<br />
Neotropics: Ecological, Taxonomic and Evolutionary<br />
Perspectives. Commemorative Volume of<br />
the 70th Anniversary of the Institute de Biologia,
AGUIRRE-MACEDO ET AL.—ENDOHELMINTHS FROM NICARAGUAN FISHES 195<br />
Universidad Nacional Autonoma de Mexico.<br />
Mexico, D.F., Mexico.<br />
J. C. Chubb, and G. C. Pavanelli. 1999.<br />
Cestodes in South American freshwater teleost<br />
fishes: keys to the genera and brief descriptions.<br />
Revista Brasileira de Zoologia 16:299-367.<br />
Salgado-Maldonado, G., R. Pineda-Lopez, V. M.<br />
Vidal-Martinez, and C. R. Kennedy. 1997. A<br />
checklist of metazoan parasites of cichlid fish<br />
from Mexico. Journal of the Helminthological Society<br />
of Washington 64:195-207.<br />
Scholz, T., and J. Vargas-Vazquez. 1998. Trematodes<br />
parasitizing fishes of the Rio Hondo River<br />
and freshwater lakes of Quintana Roo, Mexico.<br />
Journal of the Helminthological Society of Washington<br />
65:91-95.<br />
, , F. Moravec, C. Vivas-Rodriguez,<br />
and E. F. Mendoza-Franco. 1995. Cenotes (sinkholes)<br />
of the Yucatan Peninsula of Yucatan, Mexico,<br />
as habitat of adult trematodes of fish. Folia<br />
Parasitologica 42:37-47.<br />
, , , , and . 1996.<br />
Cestoda and Acanthocephala of fishes from cenotes<br />
( = sinkholes) of Yucatan, Mexico. Folia Parasitologica<br />
43:141-152.<br />
Szidat, L. 1954. Trematodos nuevos de peces de agua<br />
dulce de la Republica Argentina y un intento para<br />
alcarar su caracter marino. Revista del Institute de<br />
Investigacion de las Ciencias Naturales y Museo<br />
Argentine de Ciencias Naturales "Bernardino Rivadavia"<br />
3(1): 1-85.<br />
Thatcher, V. E. 1991. Amazon fish parasites. Amazoniana<br />
11:263-572.<br />
Travassos, L., J. F. Teixeira de Freitas, and A.<br />
Kohn. 1969. Trematodeos do Brasil. Memorias do<br />
Institute Oswaldo Cruz 67:1-886.<br />
Vidal-Martinez, V. M., M. L. Aguirre-Macedo, T.<br />
Scholz, D. Gonzalez-Solis, and E. F. Mendoza-<br />
Franco. <strong>2001</strong>. Atlas of the Helminth Parasites of<br />
Cichlid Fish of Mexico. Academia, Prague, Czech<br />
Republic. 165 pp.<br />
, and C. R. Kennedy. 2000. Zoogeographical<br />
determinants of the helminth fauna composition<br />
of neotropical cichlid fish. Pages 227-290 in A.<br />
N. Garcfa-Aldrete, G. Salgado-Maldonado, and V.<br />
M. Vidal-Martinez, eds. Metazoan Parasites in the<br />
Neotropics: Ecological, Taxonomic and Evolutionary<br />
Perspectives. Commemorative Volume of<br />
the 70th Anniversary of the Institute de Biologia,<br />
Universidad Nacional Autonoma de Mexico.<br />
Mexico, D.F., Mexico.<br />
T. Scholz, and M. L. Aguirre-Macedo.<br />
<strong>2001</strong>. Dactylogyridae of cichlid fishes from Nicaragua,<br />
Central America, with descriptions of<br />
Gussevia herotilapiac and three new species of<br />
Sciadicleithrum (Monogenea: Ancyrocephalinae).<br />
<strong>Comparative</strong> <strong>Parasitology</strong> <strong>68</strong>:76-86.<br />
Watson, D. E. 1976. Digenea of fishes from Lake Nicaragua.<br />
Pages 251-260 in T. B. Thorson, ed. Investigations<br />
of the Ichthyofauna of Nicaraguan<br />
Lakes. School of Life Sciences, University of Nebraska,<br />
Lincoln, Nebraska, U.S.A.<br />
Relocation of the Onderstepoort Helminthological Collection<br />
We have been notified that the Onderstepoort Helminthological Collection has moved and been renamed,<br />
and is now under the curatorship of Professor J. Boomker, Department of Veterinary Tropical Diseases,<br />
and Dr. E. van den Berg, Plant Protection Unit, University of Pretoria, Private Bag X04, Onderstepoort,<br />
0110 South Africa. The collection is now known as the National Collection of Animal Helminths and is<br />
fully accessible. Prospective lenders, or those seeking further information, can notify Professor Boomker<br />
at the above address, phone: +27-12-529-8166, fax: +27-12-529-8312, or e-mail: jboomker@op.up.ac.za.<br />
Copyright © 2011, The Helminthological Society of Washington
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 196-203<br />
Helminth Parasites of Freshwater Fishes of the Balsas River<br />
Drainage Basin of Southwestern Mexico<br />
GUILLERMO SALGADO-MALDONADO,1'4 GUILLERMINA CABANAS-CARRANZA,1<br />
JUAN M. CASPETA-MANDUJANO,2 EDUARDO SoTO-GALERA,3 ELIZABETH MAYEN-PENA,'<br />
DAVID BRAILOVSKY,' AND RAFAEL BAEZ-VALE'<br />
1 Institute de Biologia, Universidad Nacional Autonoma de Mexico, Apartado Postal 70-153, CP 04510,<br />
Mexico (e-mail: gsalgado@mail.ibiologia.unam.mx),<br />
2 Centre de Investigaciones Biologicas, Universidad Autonoma del Estado de Morelos, Av. Universidad 1001,<br />
CP 62210, Cuernavaca, Morelos, Mexico, and<br />
3 Laboratorio de Ictiologfa y Limnologia, Escuela Nacional de Ciencias Biologicas, Institute Politecnico<br />
Nacional, Carpio y Plan de Ayala, Santo Tomas, Mexico, D.F., Mexico<br />
ABSTRACT: This study presents the results of the first survey of the helminth parasites in fishes in the Balsas<br />
River drainage, southwestern Mexico. A total of 25 species of helminth parasites in 13 freshwater fish species<br />
(n = 1,045) was collected between December 1995 and September 1998. The most prevalent and widespread<br />
helminth species was the Asian tapeworm Bothriocephalus acheilognathi. Two features characterize the helminth<br />
fauna of the Balsas River basin fishes: (1) a predominance of nematode and trematode species coupled with a<br />
scarcity of monogeneans and acanthocephalans; and (2) all helminths found had previously been reported from<br />
other regions of Mexico; therefore the composition of the helminth fauna of the fishes of the Balsas River<br />
drainage is not very distinct from that of fishes from other previously studied freshwater basins in Mexico.<br />
KEY WORDS: Monogenea, Digenea, Cestoda, Nematoda, Acanthocephala, freshwater fish, Balsas River, southwestern<br />
Mexico, survey.<br />
The Balsas River basin is the largest river<br />
drainage in southwestern Mexico. The Balsas<br />
River has its source at about 3,660 m altitude in<br />
the Sierra Madre del Sur. It flows generally eastwest<br />
through the states of Tlaxcala, Puebla,<br />
Guerrero, and Michoacan and receives several<br />
major inflows from the states of Oaxaca, Mexico,<br />
Morelos, and Jalisco before emptying into<br />
the Pacific Ocean. This river has a fish fauna<br />
composed of 37 species of 26 genera in 10 families.<br />
In addition to native fishes, exotic species<br />
such as Asian cyprinids (carps) and African<br />
cichlids (tilapias) have been introduced into<br />
many areas.<br />
Little information exists about the occurrence<br />
of helminth parasites of fishes from the Balsas<br />
River (Osorio-Sarabia, 1982, 1984; Salgado-<br />
Maldonado et al., 1998; Caspeta-Mandujano and<br />
Moravec, 2000; Caspeta-Mandujano et al., 2000;<br />
Moravec, 2000; Moravec et al., 2000), and the<br />
present report is the first survey of the helminth<br />
parasites of fishes of this drainage system. The<br />
aim of this work is to report the survey results,<br />
and the distribution and intensity data for these<br />
helminth parasites.<br />
Corresponding author.<br />
196<br />
Copyright © 2011, The Helminthological Society of Washington<br />
Materials and Methods<br />
A total of 1,045 fish was collected from 28 sites,<br />
mostly rivers, in the Balsas River basin between December<br />
1995 and September 1998 (Table 1, Fig. 1).<br />
Fish at each site were captured by electrofishing or<br />
by gill nets. Live fish were brought to the laboratory<br />
and examined within 48 hr after capture using standard<br />
procedures. The following fishes were examined (* indicates<br />
species endemic to the Balsas River basin): Cyprinidae—*Hybopis<br />
boucardi (Giinther, 18<strong>68</strong>) (Balsas<br />
shiner, n= 111); Characidae—Astyanax fasciatus (Cuvier,<br />
1819) (Mexican tetra, n = 166); Ictaluridae—*Ictalurus<br />
balsanus (Jordan and Snyder, 1899) (Balsas<br />
catfish, n = 1); Ictalurus punctatus (Rafinesque, 1818)<br />
(channel catfish, n = 2); Goodeidae—Goodca atripinnis<br />
Jordan, 1880 (blackfin goodea, n = 6); *Ilyodon<br />
whitei (Meek, 1904) (Balsas splitfin, n = 59); Poeciliidae—Heterandria<br />
bimaculatiis (Heckel, 1848) (spottail<br />
killifish, n = 88), Poecilia reticulata Peters, 1860<br />
(guppy, n — 20), Poecilia sphenops Valenciennes in<br />
Cuvier and Valenciennes, 1846 (Mexican molly, n =<br />
261), Poeciliopsis gracilis (Heckel, 1848) (porthole<br />
livebearer, n = 156), Poeciliopsis infans (Woolman,<br />
1894) (Lerma livebearer, n = 20); Cichlidae—*Cichlasoma<br />
istlanum (Jordan and Snyder, 1899) (redside<br />
cichlid, n = 32), Cichlasoma nigrofasciatum (Giinther,<br />
1867) (convict cichlid, n = 123). Fish sample sizes per<br />
site are given in Table 1.<br />
All helminths recovered from each fish were counted.<br />
Digeneans (adults and metacercariae), cestodes,<br />
and nematodes were fixed in hot 10% neutral formalin.<br />
Acanthocephalans were placed in distilled water and<br />
refrigerated overnight (6-12 hr) to evert the proboscis,
Table 1. Helminths found in freshwater fishes from the Balsas River basin, Mexico.<br />
^Locality (no<br />
total no. of helm<br />
Species Site Hosts<br />
Monogenea<br />
Gyrodactylus sp. Gills<br />
P. gracilis PL (2/18; 7; 0.4 ± 1.1 [3-4])<br />
P. infans VJ (1/20; 1)<br />
A.fasciatus PL (3/13; 12; 0.9 ± 1.8 [3-5]),<br />
[1-2]), PT (5/10; 30; 3.0 ± 3<br />
Urocleidoides cf. costaricensis (Price Gills<br />
and Bussing, 1967) Kritsky and<br />
Leiby, 1972<br />
/. whitei CH (1/22; 3)<br />
P. sphenops AR (1/7; 1), CH (2/43; 2; 0.05<br />
P. gracilis AR (9/11; 17; 1.5 ± 1.1 [1-4])<br />
A. fasciatus CU (1/11; 1)<br />
P. reticulata AC (3/20; 3; 0.2 ± 0.4 [1-1])<br />
P. sphenops HA (2/15; 16; 1.1 ± 3.9 [1-15]<br />
H. boucardi JU (1/8; 1), IX (5/10; 18; 1.8 ±<br />
Trematoda<br />
Saccocoelioides sogandaresi Lums- Intestine<br />
den, 1961<br />
Magnivitellinum simplex Kloss, 1966 Intestine<br />
Diplostomum cf. compaction (Lutz, Eyes, brain, body cavity<br />
1928)<br />
Posthodiplostomum minimum Muscle, liver, eyes,<br />
(MacCallum, 1921) mesentery, body cavity<br />
G. atripinnis JU (6/6; 801; 134.0 ± 37.5 [83<br />
H. bimacidata HJ (1/25; 1)<br />
P. sphenops HA (5/15; 39; 2.6 ± 6.0 [1-22]<br />
1), AM (1/16; 1); OT (1/2; 1<br />
P. infans VJ (19/20; 307; 15.4 ± 14.3 [3<br />
C. istlanum AM (1/4; 2)<br />
C. nigrofasciatum CO (5/44; 5; 0.1 ± 0.3 [1-1]),<br />
AM (8/20; 25; 1.2 ± 2.3 [1-9])<br />
H. boucardi CU (5/14; 10; 0.7 ± 1.3 [1-4])<br />
A.fasciatus PL (1/13; 1), HJ (1/5; 1)<br />
P. sphenops CO (1/10; 1), HJ (10/40; 14; 0.<br />
P. gracilis PL (1/18; 2), HJ (2/3; 3; 1.0 ±<br />
C. istlanum AM (3/4; 157; 39.2 ± 53.3 [16<br />
C. nigrofasciatum CH (17/22; 125; 5.7 ± 5.7 [1-2<br />
16.6 ± 13.1 [1-55]), AM (18<br />
A. fasciatus AM (4/26; 5; 0.2 ± 0.5 [1-2])<br />
Uvulifer sp. Skin, fins<br />
A. fasciatus AM (2/26; 5; 0.2 ± 0.7 [2-3])<br />
/. whitei CH (18/22; 1926; 87.5 ± 97.4<br />
P. sphenops CH (6/43; 28; 0.6 ± 2.3 [1-13]<br />
P. gracilis CH (3/15; 4: 0.3 ± 0.6 [1-2]),<br />
C. nigrofasciatum CH (2/22; 2; 0.1 ± 0.3 [1-1]),<br />
Clinostomum complanatum (Rudol- Fins, opercula, body<br />
phi, 1814) cavity<br />
Centrocestus formosanus (Nishigori, Gills<br />
1924)<br />
Copyright © 2011, The Helminthological Society of Washington
Table 1. Continued.<br />
'•'Locality (no.<br />
total no. of helmi<br />
Species Site Hosts<br />
H. boucardi CU (15/19; 103; 5.4 ± 10.8 [1-4<br />
[1-6]), IX (1/10; 1), PT (7/7;<br />
(5/15; 8; 0.5 ± 1.0 [1-4]), AM<br />
A. fasciatus CU (1/11; 1)<br />
H. bimacuiata CO ((1/13; 1), HJ (5/25; 7; 0.3 ±<br />
P. reticulata AC (1/20; 1)<br />
P. sphenops PT (1/5; 1), MI (1/15; 2), XO (3<br />
0.4 ± 0.9 [1-3]), PL (2/13; 2;<br />
5; 1)<br />
P. gracilis CO (2/19; 2; 0.1 ± 0.3 [1-1]), T<br />
C. istlamim TE (1/11; 2)<br />
C. nigrofasciatum HJ (3/21; 5; 0.2 ± 0.7 [1-3])<br />
P. sphenops HA ((2/15; 2; 0.1 ± 0.4 [1-1]), T<br />
XO (3/16; 6; 0.4 ± 0.8 [2-2])<br />
P. gracilis TE (9/16; 15; 0.9 ± 1.1 [1-3])<br />
A. fasciatus CU (1/11; 1)<br />
P. gracilis TE (1/16; 1)<br />
C. istlamim TE (3/11; 11; 1.0 ± 2.6 [1-9])<br />
Intestine<br />
Cestoda<br />
Bothriocephalus archeilognathi Yamaguti,<br />
1934<br />
Body cavity, mesentery,<br />
liver<br />
Glossocercus auritus (Rudolphi,<br />
1819) Bona, 1994<br />
P. sphenops TE (2/6; 18; 3.0 ± 6.0 [3-15])<br />
P. gracilis TE (4/16; 19; 1.2 ± 2.4 [2-8])<br />
Parvitaenia cochlearii Coil, 1955 Liver<br />
Parvitaenia macropeos (Wedl, 1855) Liver<br />
Baer and Bona, 1960<br />
Valipora minuta (Coil, 1950) Baer Liver, gall bladder<br />
and Bona, 1960<br />
A. fasciatus PL (1/13; 5)<br />
P. sphenops CH (7/43; 42; 1.0 ± 2.7 [1-10]),<br />
± 4.5 [1-13]), HJ (5/40; 116;<br />
C. nigrofasciatum CH (1/22; 1)<br />
H. boucardi JU (5/8; 15; 1.9 ± 2.1 ([1-6]), C<br />
1.7 [2-4]), RP (6/7; 80; 11.4 ±<br />
0.3 ± 0.5 [1-1]), MI (7/10; 34<br />
C. istlamim CO (1/1; 4), AM (4/4; 143; 35.8<br />
C. nigrofasciatum AR (8/16; 35; 2.2 ± 3.6 [1-14]),<br />
1.0 [1-6]), HJ (17/21; 99; 4.7<br />
C. atripinnis JU (5/6; 33; 5.5 ± 4.9 [1-13])<br />
Nematoda<br />
Capillaria cyprinodonticola Huffman Intestine,<br />
and Bullock, 1973 liver<br />
Rhabdochona canademis Moravec Intestine<br />
and Arai, 1971<br />
Rhabdochona kidderi Pearse, 1936 Intestine<br />
A. fasciatus CO (2/13; 2; 0.1 ± 0.4 [0-1]), P<br />
0.2 ± 0.4 [1-1]), PT (1/10; 1)<br />
Rhabdochona lichtenfelsi Sanchez- Intestine<br />
Alvarez et al., 1998<br />
Rhabdochona mexicana Caspeta- Intestine<br />
Mandujano et al., 2000<br />
Copyright © 2011, The Helminthological Society of Washington
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200 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
MEXICO ,-•* ! \TLAXCAl_A<br />
Figure 1. The Balsas River drainage basin of southwestern Mexico, showing the fish collection sites.<br />
Double circles indicate sites where no infected fish were collected.<br />
then fixed in hot 10% formalin. Digeneans, cestodes,<br />
and acanthocephalans were stained with Mayer's paracarmine<br />
or Ehrlich's hematoxylin, dehydrated through<br />
a graded alcohol series, cleared in methyl salicylate,<br />
and whole-mounted. Nematodes were cleared with<br />
glycerine for light microscopy and stored in 70% ethanol.<br />
Voucher specimens of all taxa have been deposited<br />
in the Coleccion Nacional de Helmintos, Institute<br />
de Biologfa, Universidad Nacional Autonoma de Mexico.<br />
Infection parameters utilized are those proposed<br />
by Margolis et al. (1982), that is, prevalence (% infected)<br />
and abundance of infection (number of parasites<br />
per examined fish), expressed as mean ± standard<br />
deviation, followed by the range of intensity.<br />
Results<br />
The parasites encountered, their hosts, collection<br />
locations, infection sites, and prevalence,<br />
abundance, and range of intensity of helminth<br />
species are summarized in Table 1.<br />
Only 18 of the 1,045 host fish examined harbored<br />
monogeneans. Eight Gyrodactylus sp.<br />
were collected from 2 P. gracilis and 1 P. infans.<br />
Fifteen A. fasciatus were found to harbor<br />
66 Urocleidoides cf. costaricensis.<br />
Only 36 (3.4%) of the necropsied fish (3 species)<br />
were parasite-free. These fish included both<br />
ictalurid species, and 33 /. whitei from CR. Fish<br />
at 8 of the 28 collection sites sampled were not<br />
Copyright © 2011, The Helminthological Society of Washington<br />
infected at all: HU, PE, CI, PA, XA, TL, AH,<br />
and CR (Fig. 1).<br />
The most prevalent and widespread helminth<br />
parasite was the cestode Bothriocephalus acheilognathi<br />
that was recorded in 8 of the 13 Balsas<br />
River fish species, at infection intensities<br />
from 1 to 46.<br />
Discussion<br />
Data from this survey provide further evidence<br />
to support Moravec's (1998) contention<br />
that nematodes represent a significant component<br />
of helminth faunas in tropical freshwater<br />
fishes. They also corroborate the statement of<br />
Salgado-Maldonado and Kennedy (1997) that<br />
richness in digenean species is a characteristic<br />
of these helminth communities. In contrast,<br />
acanthocephalans were found to be very rare in<br />
the Balsas River survey, supporting the claim of<br />
Salgado-Maldonado et al. (1992) that adult<br />
acanthocephalans are generally very rare parasites<br />
in Mexican freshwater fishes. Adult cestodes<br />
are not common parasites in Mexican<br />
freshwater fishes; however, this survey found 4<br />
metacestode species. Previous surveys from<br />
most other geographical areas in Mexico (Pine-
da-Lopez et al., 1985; Leon, 1992; Jimenez-<br />
Garcia, 1994; Salgado-Maldonado et al., 1997)<br />
did not reveal a rich fauna of cestodes (but see<br />
Scholz et al., 1996). Monogeneans have only exceptionally<br />
been reported from freshwater fishes<br />
in Mexico (Lamothe-Argumedo, 1981), but a<br />
number of species have been found recently, in<br />
particular in southeastern Mexico (Kritsky et al.,<br />
1994, 2000; Mendoza-Franco et al., 1997,<br />
1999).<br />
Most of the parasites recorded in this survey<br />
are shared with freshwater fishes inhabiting other<br />
Mexican drainage basins (see Pineda-Lopez<br />
et al., 1985; Jimenez-Garcia, 1994; Moravec,<br />
Vivas-Rodriguez, Scholz, Vargas-Vazquez,<br />
Mendoza-Franco, and Gonzalez-Soli's, 1995;<br />
Moravec, Vivas-Rodriguez, Scholz, Vargas-Vazquez,<br />
Mendoza-Franco, Schmitter-Soto, and<br />
Gonzalez-Soli's, 1995; Scholz et al., 1995, 1996;<br />
Salgado-Maldonado et al., 1997; Moravec,<br />
1998; Moravec et al., 2000; Scholz and Vargas-<br />
Vazquez, 1998; Scholz and Salgado-Maldonado,<br />
2000). Six adult species are of neotropical origin:<br />
Urocleidoid.es cf. costaricensis, M. simplex,<br />
R. kidderi, R. lichtenfelsi, R. mexicana, and N.<br />
golvani. Saccocoelioides sogandaresi, Rhabdochona<br />
canadensis, and C. cyprinodonticola have<br />
been recorded in various freshwater fishes in<br />
Canada and southern North America (Lumsden,<br />
1963; Moravec and Aral, 1971; Moravec, 1998).<br />
Twelve of 25 helminth species recorded during<br />
this survey were larval forms that utilized<br />
small freshwater fishes as intermediate hosts. All<br />
these allogenic species are widespread taxa, with<br />
wide distributions within Mexico and broad host<br />
specificity. Thus, they can be regarded as an<br />
ecological component of the fish parasite communities<br />
in the Balsas River basin. The metacercariae<br />
of C. complanatum, P. minimum, and<br />
Diplostomum cf. compactum, as well as the larvae<br />
of nematodes Eustrongylides sp., Contracaecum<br />
sp., and Acuariidae gen. sp. have been<br />
commonly recorded in cichlids, poeciliids, characiids,<br />
pimelodids, and other fish families from<br />
southern Mexico (Pineda-Lopez, 1985; Pineda-<br />
Lopez et al., 1985; Osorio-Sarabia et al., 1987;<br />
Jimenez-Garcia, 1994; Moravec, Vivas-Rodriguez,<br />
Scholz, Vargas-Vazquez, Mendoza-Franco,<br />
Schmitter-Soto, and Gonzalez-Solis, 1995;<br />
Scholz et al., 1995; Salgado-Maldonado et al.,<br />
1997). They have also been reported in atherinids,<br />
goodeids, and other fish families from the<br />
Lerma Santiago River basin in the highland pla-<br />
SALGADO-MALDONADO ET AL.—HELMINTHS OF MEXICAN FISHES 201<br />
teau of central Mexico (Osorio-Sarabia et al.,<br />
1986; Salgado-Maldonado and Osorio-Sarabia,<br />
1987; Leon, 1992; Peresbarbosa et al., 1994).<br />
All these helminth species are widely distributed<br />
in North America, and some are worldwide<br />
(Hoffman, 1967; Yamaguti, 1971; Gibson,<br />
1996).<br />
Some of the helminths found have been introduced<br />
to Mexico with exotic fish or other animals.<br />
The Asian fish tapeworm (B. acheilognathi)<br />
has been disseminated globally in association<br />
with Asian cyprinids (grass and common<br />
carp) introduced to several countries for use in<br />
aquaculture (Salgado-Maldonado et al., 1986).<br />
This tapeworm has broad host specificity and<br />
now occurs in more than 15 freshwater fish species<br />
in Mexico (Garcia and Osorio-Sarabia,<br />
1991). We found B. acheilognathi widely distributed<br />
within the Balsas River basin, parasitizing<br />
8 fish species, mainly poeciliids.<br />
Another example is the heterophyid trematode<br />
C. formosanus that was introduced into Mexico<br />
most probably with the imported thiarid snail<br />
Melanoides tuberculata (Miiller, 1774) serving<br />
as the first intermediate host. This trematode has<br />
rapidly spread to an extensive area, including<br />
central Mexico and both the Atlantic and Pacific<br />
coasts, apparently aided by the previous expansion<br />
of M. tuberculata within Mexico. The<br />
metacercariae of C. formosanus are encysted in<br />
the gills of a wide spectrum of native fishes including<br />
members of Atherinidae, Cichlidae, Cyprinidae,<br />
Eleotridae, Goodeidae, Ictaluridae, and<br />
Poeciliidae (see Scholz and Salgado-Maldonado,<br />
2000). The adults are parasites in piscivorous<br />
birds and mammals. An increasing number of<br />
recent records of C. formosanus in numerous<br />
new hosts and regions, including the Balsas River<br />
drainage, suggests that this helminth is continuing<br />
to expand its distribution (Scholz and<br />
Salgado-Maldonado, 2000).<br />
Too few studies have been undertaken to draw<br />
conclusions about the zoogeographic characteristics<br />
of the helminth communities in the freshwater<br />
fish species of the Balsas River basin.<br />
However, two general statements can be made<br />
about these faunas. The first is that nematode<br />
and trematode species predominate, with only a<br />
few monogeneans and acanthocephalans being<br />
present. Second, all helminths found had previously<br />
been reported from other regions of Mexico;<br />
therefore the taxonomic composition of the<br />
helminth fauna of the fishes of the Balsas River<br />
Copyright © 2011, The Helminthological Society of Washington
202 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
drainage is not very distinct from that seen in<br />
other previously studied freshwater basins in<br />
Mexico.<br />
Acknowledgments<br />
This study was supported by project no.<br />
400355-5-276<strong>68</strong>N from the Consejo Nacional<br />
para la Ciencia y la Tecnologia (CONACyT),<br />
Mexico and by the Comision Nacional para el<br />
Conocimiento y Uso de la Biodiversidad (CON-<br />
ABIO), Mexico, project nos. H007 and L051.<br />
We are indebted to Drs. Frantisek Moravec and<br />
Tomas Scholz for identification of nematodes<br />
and cestodes. Thanks are also due Aitzane Delgado-Yoshino,<br />
Erick Alcantara, Alejandra Hernandez-Rodriguez,<br />
Nancy Minerva Lopez-Flores,<br />
Isabel Cristina Caneda-Guzman, Norman<br />
Mercado-Silva, and Felipe Villegas-Marquez for<br />
their assistance in the field and laboratory.<br />
Literature Cited<br />
Caspeta-Mandujano, J. M., and F. Moravec. 2000.<br />
Two new intestinal nematodes of Profundulus labialis<br />
(Pisces, Cyprinodontidae) from fresh waters<br />
in Mexico. Acta Parasitologica 45:332-339.<br />
, , M. A. Delgado-Yoshino, and G.<br />
Salgado-Maldonado. 2000. Seasonal variations<br />
in the occurrence and maturation of the nematode<br />
Rhabdochona kidderi in Cichlasoma nigrofasciatum<br />
of the Amacuzac River, Mexico. Helminthologia<br />
37:29-33.<br />
Garcia, P. L., and D. Osorio-Sarabia. 1991. Distribucion<br />
actual de Bothriocephalus acheilognathi<br />
en Mexico. Anales del Institute de Biologia, Universidad<br />
Nacional Autonoma de Mexico, Serie<br />
Zoologia 62:523-526.<br />
Gibson, D. I. 1996. Trematoda. In L. Margolis and Z.<br />
Kabata, eds. Guide to the Parasites of Fishes of<br />
Canada. Part IV. Canadian Special Publications of<br />
Fisheries and Aquatic Sciences 124. 373 pp.<br />
Hoffman, G. L. 1967. Parasites of North American<br />
Freshwater Fishes. University of California Press,<br />
Berkeley, California, U.S.A. 486 pp.<br />
Jimenez-Garcia, M. I. 1994. Fauna helmintologica de<br />
Cichlasoma fenestratum (Pisces: Cichlidae) del<br />
lago de Catemaco, Veracruz, Mexico. Anales del<br />
Institute de Biologia, Universidad Nacional Autonoma<br />
de Mexico, Serie Zoologia 64:75-78.<br />
Kritsky, D. C., E. F. Mendoza-Franco, and T.<br />
Scholz. 2000. Neotropical Monogenoidea. 36.<br />
Dactylogyrids from the Gills of Rhamdia guatemalensis<br />
(Siluriformes: Pimelodidae) from Cenotes<br />
of the Yucatan Peninsula, Mexico, with Proposal<br />
of Ameloblastella gen. n. and Aphanoblastella<br />
gen. n. (Dactylogyridae: Ancyrocephalinae).<br />
<strong>Comparative</strong> <strong>Parasitology</strong> 67:76-84.<br />
, V. M. Vidal-Martinez, and R. Rodriguez-<br />
Canul. 1994. Neotropical Monogenoidea 19. Dactylogyridae<br />
of cichlids (Perciformes) from the Yucatan<br />
Peninsula, with descriptions of three new<br />
Copyright © 2011, The Helminthological Society of Washington<br />
species of Sciadicleithriim Kritsky, Thatcher and<br />
Boeger, 1989. Journal of the Helminthological Society<br />
of Washington 61:26-33.<br />
Lamothe-Argumedo, R. 1981. Monogeneos parasitos<br />
de peces. VIII. Descripcion de una nueva especie<br />
del genero Octomacrum Mueller, 1934 (Monogenea:<br />
Discocotylidae). Anales del Instituto de Biologia,<br />
Universidad Nacional Autonoma de Mexico,<br />
Serie Zoologia 51:51—60.<br />
Leon, R. V. 1992. Fauna helmintologica de algunos<br />
vertebrados acuaticos de la cienega de Lerma, Estado<br />
de Mexico. Anales del Instituto de Biologia,<br />
Universidad Nacional Autonoma de Mexico, Serie<br />
Zoologia 63:151-153.<br />
Lumsden, R. D. 1963. Saccocoelioides sogandaresi<br />
sp. n., a new haploporid trematode from the sailfin<br />
molly Mollienisia latipinna Le Sueur in Texas.<br />
Journal of <strong>Parasitology</strong> 49:281-284.<br />
Margolis, L., G. W. Esch, J. C. Holmes, A. M. Kuris,<br />
and G. A. Schad. 1982. The use of ecological<br />
terms in parasitology (report of an ad hoc committee<br />
of the American Society of Parasitologists).<br />
Journal of <strong>Parasitology</strong> <strong>68</strong>:131-133.<br />
Mendoza-Franco, E. F., T. Scholz, and V. M. Vidal-<br />
Martinez. 1997. Sciadicleithriim meekii sp. n.<br />
(Monogenea: Ancyrocephalinae) from the gills of<br />
Cichlasoma meeki (Pisces: Cichlidae) from cenotes<br />
(=sinkholes) of the Yucatan Peninsula, Mexico.<br />
Folia Parasitologica 44:205-208.<br />
, , C. Vivas-Rodriguez, and J. Vargas-<br />
Vazquez. 1999. Monogeneans of freshwater fishes<br />
from cenotes (sinkholes) of the Yucatan Peninsula,<br />
Mexico. Folia Parasitologica 46:267-273.<br />
Moravec, F. 1998. Nematodes of Freshwater Fishes<br />
of the Neotropical Region. Academia, Prague,<br />
Czech Republic. 464 pp.<br />
—. 2000. Systematic status of Laurotravassoxyuris<br />
bravoae Osorio-Sarabia, 1984 (Nematoda:<br />
Pharyngodonidae) [=Atractis bravoae (Osorio-<br />
Sarabia, 1984) n. comb.: Cosmocercidae]. Systematic<br />
<strong>Parasitology</strong> 46:117-122.<br />
, and H. P. Arai. 1971. The North and Central<br />
American species of Rhabdochona Railliet, 1916<br />
(Nematoda: Rhabdochonidae) of fishes, including<br />
Rhabdochona canadensis sp. nov. Journal of the<br />
Fisheries Research Board of Canada 28:1645-<br />
1662.<br />
, G. Salgado-Maldonado, and J. M. Caspeta-<br />
Mandujano. 2000. Rhabdochona mexicana sp. n.<br />
(Nematoda: Rhabdochonidae) from the intestine<br />
of characid fishes in Mexico. Folia Parasitologica<br />
47:211-215.<br />
, C. Vivas-Rodriguez, T. Scholz, J. Vargas-<br />
Vazquez, E. Mendoza-Franco, and D. Gonzalez-Solis.<br />
1995. Nematodes parasitic in fishes of<br />
cenotes (=sinkholes) of the Peninsula of Yucatan,<br />
Mexico. Part 1. Adults. Folia Parasitologica 42:<br />
115-129.<br />
, , , , , J- J-<br />
Schmitter-Soto, and D. Gonzalez-Solis. 1995.<br />
Nematodes parasitic in fishes of cenotes ( = sinkholes)<br />
of the Peninsula of Yucatan, Mexico. Part<br />
2. Larvae. Folia Parasitologica 42:199-210.<br />
Osorio-Sarabia, D. 1982. Descripcion de una nueva
especie del genero Goezici Zeder, 1800 (Nematoda:<br />
Goeziidae) en peces de agua dulce de Mexico.<br />
Anales del Instituto de Biologfa, Universidad Nacional<br />
Autonoma de Mexico, Serie Zoologia 52:<br />
71-87.<br />
. 1984. Descripcion de una especie nueva del<br />
genero Laurotravassoxyuris Vigueras, 1938<br />
(Nematoda: Syphaciidae) en peces de agua dulce<br />
de Mexico. Anales del Instituto de Biologfa, Universidad<br />
Nacional Autonoma de Mexico, Serie<br />
Zoologia 54:23-33.<br />
, G. Perez, and G. Salgado-Maldoiiado.<br />
1986. Helmintos de peces del lago de Patzcuaro,<br />
Michoacan I: Helmintos de Chirostoma estor el<br />
"pescado bianco." Taxonomfa. Anales del Instituto<br />
de Biologfa, Universidad Nacional Autonoma<br />
de Mexico, Serie Zoologfa 57:61-92.<br />
-, R. Pineda-Lopez, and G. Salgado-Maldonado.<br />
1987. Fauna helmintologica de peces dulceacufcolas<br />
de Tabasco. Estudio preliminar. Universidad<br />
y Ciencia 4:5-31.<br />
Peresbarbosa, R. E., G. Perez, and L. Garcia-Prieto.<br />
1994. Helmintos parasites de tres especies de<br />
peces (Goodeidae) del lago de Patzcuaro, Michoacan.<br />
Anales del Instituto de Biologfa, Universidad<br />
Nacional Autonoma de Mexico, Serie<br />
Zoologfa 65:201-204.<br />
Pineda-Lopez, R. 1985. Infeccion por metacercarias<br />
(Platyhelminthes: Trematoda) en peces de agua<br />
dulce de Tabasco. Universidad y Ciencia 2:47-60.<br />
, V. M. Carballo-Cruz, M. G. Fucugauchi,<br />
and L. Garcia-Magana. 1985. Metazoarios parasitos<br />
de peces de importancia comercial en la region<br />
de Los Rfos, Tabasco, Mexico. Pages 197-<br />
270 in Usumacinta: Investigacion Cientffica en la<br />
Cuenca del Usumacinta. Gobierno del Estado de<br />
Tabasco, Mexico.<br />
Salgado-Maldonado, G., G. Cabanas-Carranza,<br />
and J. M. Caspeta-Mandujano. 1998. Crcptotrema<br />
agonostomi n. sp. (Trematoda: Allocreadiidae)<br />
from the intestine of freshwater fish of Mexico.<br />
Journal of <strong>Parasitology</strong> 84:431-434.<br />
, S. Guillen-Hernandez, and D. Osorio-Sarabia.<br />
1986. Presencia de Bothriocephalus acheilognathi<br />
Yamaguti, 1934 (Ccstoda: Bothriocephal-<br />
SALGADO-MALDONADO ET AL.—HELMINTHS OF MEXICAN FISHES 203<br />
idae) en peces de Patzcuaro, Michoacan, Mexico.<br />
Anales del Instituto de Biologfa, Universidad Nacional<br />
Autonoma de Mexico, Serie Zoologfa 57:<br />
213-218.<br />
, M. I. Jimenez-Garcia, and V. Leon-Regagnon.<br />
1992. Presence of Octospiniferoides chandleri<br />
Bullock, 1957 in Heterandria bimaculata<br />
from Catemaco Veracruz, and considerations<br />
about the acanthocephalans of freshwater fishes of<br />
Mexico. Memorias do Instituto Oswaldo Cruz<br />
87(supplement 1):239-240.<br />
, and C. R. Kennedy. 1997. Richness and similarity<br />
of helminth communities in the tropical<br />
cichlid fish Cichlasoma umphthalmus from the<br />
Yucatan Peninsula, Mexico. <strong>Parasitology</strong> 114:<br />
581-590.<br />
, and D. Osorio-Sarabia. 1987. Helmintos de<br />
algunos peces del lago de Patzcuaro. Ciencia y<br />
Desarrollo 13:41-57.<br />
, R. Pineda-Lopez, V. M. Vidal-Martinez,<br />
and C. R. Kennedy. 1997. A checklist of metazoan<br />
parasites of cichlid fish from Mexico. Journal<br />
of the Helminthological Society of Washington<br />
64:195-207.<br />
Scholz, T., and G. Salgado-Maldonado. 2000. The<br />
introduction and dispersal of Centrocestus formosanus<br />
(Nishigori, 1924) (Digenea: Heterophyidae)<br />
in Mexico: a review. American Midland Naturalist<br />
143:185-200.<br />
, and J. Vargas-Vazquez. 1998. Trematodes<br />
from fishes of the Rio Hondo River and freshwater<br />
lakes of Quintana Roo, Mexico. Journal of the<br />
Helminthological Society of Washington 65:91-<br />
95.<br />
, , F. Moravec, C. Vivas-Rodriguez,<br />
and E. Mendoza-Franco. 1995. Metacercariae of<br />
trematodes of fishes from cenotes ( = sinkholes) of<br />
the Yucatan Peninsula, Mexico. Folia Parasitologica<br />
42:173-192.<br />
, , , , and . 1996.<br />
Cestoda and Acanthocephala of fishes from cenotes<br />
( = sinkholes) of Yucatan, Mexico. Folia Parasitologica<br />
43:141-152.<br />
Yamaguti, S. 1971. Synopsis of Digenetic Vertebrates<br />
of Vertebrates. Vol. I. Keigaku Publishing Company,<br />
Tokyo, Japan. 1,074 pp.<br />
Copyright © 2011, The Helminthological Society of Washington
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 204-218<br />
A Checklist of Helminth Parasites of Freshwater Fishes from the<br />
Lerma-Santiago River Basin, Mexico<br />
GUILLERMO SALGADO-MALDONADO,1-4 GUILLERMINA CABANAS-CARRANZA,1 EOUARDO SOTO-<br />
GALERA,2 JUAN M. CASPETA-MANDUJANO,3 R. GRISELDA MORENO-NAVARRETE,1<br />
PETRA SANCHEZ-NAVA,' AND ROGELIO AGUILAR-AGUILAR'<br />
1 Institute de Biologia, Universidad Nacional Autonoma de Mexico, Apartado Postal 70-153, CP 04510,<br />
Mexico, D.F., Mexico (e-mail: gsalgado@mail.ibiologia.unam.mx),<br />
2 Laboratorio de Ictiologfa y Limnologfa, Escuela Nacional de Ciencias Biologicas, Institute Politecnico<br />
Nacional, Carpio y Plan de Ayala, Santo Totnas, CP 11340, Mexico D.F., Mexico, and<br />
3 Centre de Investigaciones Biologicas, Universidad Autonoma del Estado de Morelos, Avenida Universidad<br />
1001, CP 62210, Cuernavaca, Morelos, Mexico<br />
ABSTRACT: A checklist based on previously published records and original data is presented for the helminth<br />
parasites reported from 33 freshwater fish species from the Lerma-Santiago river basin, west-central Mexico.<br />
The checklist contains 43 helminth species, 6 (14%) of which are endemic to the basin. Fourteen of the 43 are<br />
allogenic species, mostly Nearctic in origin. Three species are anthropogenically introduced colonizers, of which<br />
the Asian fish tapeworm Bothriocephalus acheilognathi is the most widely distributed species in the basin. The<br />
checklist includes 75 new host records, and records of 12 localities where no previous surveys had been conducted.<br />
KEY WORDS: Digenea, Monogenea, Cestoda, Nematoda, Acanthocephala, freshwater fishes, Lerma-Santiago<br />
river basin, west-central Mexico, survey.<br />
At least 375 freshwater fish species, of which<br />
approximately 60% are endemic, occur in Mexico,<br />
and over 500 species occur if those living<br />
in estuaries and coastal lagoons are included<br />
(Miller, 1982; Espinosa-Perez, 1993). The Lerma-Santiago<br />
river basin in west-central Mexico<br />
has the highest percentage of endemism of any<br />
major river basin in Mexico, with 30 of its 42<br />
(72%) fish species found nowhere else (Espinosa-Perez,<br />
1993; Soto-Galera et al., 1998).<br />
This river basin (Fig. 1) drains much of westcentral<br />
Mexico and consists of 2 major rivers, the<br />
Lerma and the Santiago. The Lerma River basin<br />
is the most important hydrologic system of the<br />
Mexican Central Highland Plateau. It originates in<br />
the <strong>State</strong> of Mexico at an elevation of 3,000 m and<br />
flows for 700 km through the states of Queretaro,<br />
Guanajuato, Michoacan, and Jalisco before emptying<br />
into Chapala Lake at 1,500 m elevation. The<br />
Santiago River drains from Chapala Lake, flowing<br />
through the state of Jalisco to the Pacific Ocean.<br />
The fish fauna of the Lerma-Santiago river basin<br />
has long been studied by ichthyologists (Diaz-<br />
Pardo et al., 1993; Soto-Galera et al., 1998). No<br />
regional survey of the parasite fauna of these fish<br />
has been published, and the literature is scattered<br />
4 Corresponding author.<br />
204<br />
Copyright © 2011, The Helminthological Society of Washington<br />
in taxonomic papers (Flores-Barroeta, 1953; Lamothe-Argumedo,<br />
1970, 1981, 1988; Lamothe-Argumedo<br />
and Cruz-Reyes, 1972; Osorio-Sarabia et<br />
al., 1986; Salgado-Maldonado et al., 1986; Salgado-Maldonado<br />
and Osorio-Sarabia, 1987; Alarcon,<br />
1988; Alarcon and Castro-Aguirre, 1988; Garcfa-<br />
Prieto et al., 1988; Garcfa-Prieto and Osorio-Sarabia,<br />
1991; Leon-Regagnon, 1992; Peresbarbosa-<br />
Rojas et al., 1994; Perez-Ponce de Leon et al.,<br />
1994; Espinosa-Huerta et al., 1996; Mendoza-<br />
Garfias et al., 1996; Astudillo-Ramos and Soto-<br />
Galera, 1997; Pineda-Lopez and Gonzalez, 1997;<br />
Sanchez-Alvarez et al., 1998; Guzman-Cornejo<br />
and Garcfa-Prieto, 1999; Caspeta-Mandujano et<br />
al., 1999; Moravec et al., 2000, <strong>2001</strong>; Scholz and<br />
Salgado-Maldonado, 2000, <strong>2001</strong>). This paper<br />
compiles the extant information on the helminth<br />
parasites of freshwater fishes in the Lerma-Santiago<br />
river basin and includes original data derived<br />
from our own research. The species referred to in<br />
theses and scientific meetings do not constitute formal<br />
publications and are consequently not considered<br />
herein. This checklist should facilitate future<br />
research on the ecology, zoogeography, and biodiversity<br />
of this important river basin.<br />
Materials and Methods<br />
As part of an ongoing parasitological investigation into<br />
the helminth fauna of the freshwater fishes of Mexico, a
JALISCO<br />
SALGADO-MALDONADO ET AL.—HELMINTHS OF MEXICAN FISHES 205<br />
Chap '<br />
LAKE'CHAPALA<br />
Figure 1. The Lerma-Santiago River drainage basin of west-central Mexico, showing the fish collection<br />
sites. Locality codes as in Table 1.<br />
review of the literature dealing with freshwater fish helminth<br />
parasites in the entire Lerma-Santiago river basin<br />
was made. In addition, a total of 1,177 fish of 18 species<br />
(Table 1), from 11 localities in the Lerma-Santiago river<br />
basin (Table 2, Fig. 1) was examined for the presence of<br />
helminths from January to October 1997 and from January<br />
to March 1998.<br />
At each site, fish were captured using electrofishing or<br />
gill nets. The numbers of fish examined at each locality<br />
and collection data are given in the parasite-host list (Table<br />
3). After capture, the fish were taken live to the laboratory<br />
and examined within 48 hr using standard procedures.<br />
Briefly, all the external surfaces, viscera, and<br />
musculature of each fish host were examined under a<br />
stereomicroscope, and all the helminths encountered in<br />
each fish were counted. Digeneans (adults and larvae),<br />
cestodes, and nematodes were fixed in hot 4% neutral<br />
formalin. Acanthocephalans were placed in distilled water,<br />
refrigerated overnight (6—12 hr) to evert the proboscis,<br />
and then fixed in hot 10% formalin. Digeneans, cestodes,<br />
and acanthocephalans were stained with Mayer's paracarmine<br />
or Ehrlich's hematoxylin, dehydrated using a<br />
graded alcohol series, cleared in methyl salicylate, and<br />
whole mounted. Nematodes were cleared with glycerine<br />
for light microscopy and stored in 70% ethanol. Voucher<br />
specimens of all taxa have been deposited in the National<br />
Helminth Collection (Coleccion Nacional de Helmintos<br />
[CNHE]), Institute of Biology, National Autonomous<br />
University of Mexico (UNAM), Mexico City. Infection<br />
parameters utilized are those proposed by Margolis et al.<br />
(1982), that is, prevalence (% infected) and mean intensity<br />
of infection (number of parasites per examined fish).<br />
Voucher specimens of the following species, found in<br />
fish from the Lerma-Santiago river basin and deposited<br />
in the CNHE, were examined: Posthodiplostomum minimum<br />
(MacCallum, 1921) (nos. 001253, 001476,<br />
001748); Bothriocephalus acheilognathi Yamaguti, 1934<br />
(no. 000434); Proteocephalus pusillus Ward, 1910 (nos.<br />
000383-000386); Proteocephalus sp. (no. 000425); Ligula<br />
intestinalis (Linnaeus, 1758) [nos. 448(F) and<br />
449(F)], and Contracaeciim sp. [nos. 002508(F) and<br />
002253(F)].<br />
Results<br />
A host-parasite checklist is presented herein<br />
as Table 3. In this study, 43 helminth species are<br />
reported from 33 species of freshwater fishes of<br />
the Lerma-Santiago river basin, west-central<br />
Mexico. Six (14%) of the 43 species are endemic<br />
to the basin: A. mexicanum, M. bravoae, O.<br />
mexicanum, R. lichtenfelsi, Spinitectus sp., and<br />
B. nayaritensis. Fourteen are allogenic species<br />
that mature in, and are transported by, birds: C.<br />
Copyright © 2011, The Helminthological Society of Washington
206 COMPARATIVE PARASITOLOOY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Table 1. Fish species from the Lerma-Santiago river basin of west-central Mexico that were examined<br />
for helminths in 1997 and 1998.<br />
Fish species Common name<br />
Sample<br />
size (n)<br />
Cyprinidae<br />
*Algansea tincella (Valenciennes in Cuvier and Valenciennes, 1844) Spottail chub 17<br />
jCyprinux carpio Linnaeus, 1758 Common carp 45<br />
'•'"Notropis sallei (Giinther, 18<strong>68</strong>) Azteca chub 37<br />
*Yuriria alta (Jordan, 1880) Lerma chub 49<br />
Goodeidae<br />
''Girardinichthys multiradiatiis (Meek, 1904) Darkedged splitfin 503<br />
'•'•Goodea atripinnis Jordan, 1880 Blackfin goodea 143<br />
*Xcnotoca variants (Bean, 1887) Jeweled splitfin 56<br />
Poeciliidae<br />
Poecilia sphenops Valenciennes //; Cuvier and Valenciennes, 1846 Mexican molly 23<br />
*Poeciliopxis infans (Woolman, 1894) Lerma livebearer 16<br />
Poeciliopsis sp. 13<br />
Atherinidae<br />
*Atherinella crystallina (Jordan and Culver ;'/; Jordan, 1895) Blackfin silverside 48<br />
*Chirostoma humboldtianitm (Valenciennes in Cuvier and Valenciennes,<br />
1835) Shortfin silverside 46<br />
*Chirostoma jordani: Woolman, 1894 Mesa silverside 64<br />
*Chirostoma labarcae Meek, 1902 Sharpnose silverside 3<br />
*Chirostoma riojai Solorzano and Lopez, 1966 Toluca silverside 78<br />
Cichlidae<br />
Cichlasoma beani (Jordan, 1889) Sinaloan cichlid 32<br />
Centrarchidae<br />
t Lepomis macrochirus Rafinesque, 1819 Bluegill 2<br />
Gobiidac<br />
Awaous tajasica (Lichtenstein, 1822) River goby 2<br />
* Species endemic to the Lerma-Santiago river basin.<br />
t Species introduced to the Lerma-Santiago river basin.<br />
complanatum, Diplostomum sp., P. minimum, C.<br />
formosanus, L. intestinalis, C. cf. ralli, P. caballeroi,<br />
P. cf. urseus, P. cochlearii, V. campylane<br />
ristrota, V. mutabills, Eustrongylides sp.,<br />
Contracaecum sp., and P. brevis. Three species<br />
are recent, anthropogenically introduced colonizers:<br />
C. formosanus, P. tornentosa, and B. acheilognathi,<br />
which is the most widely distributed<br />
species in the basin. Twelve of the 33 fish species<br />
examined have not previously been surveyed<br />
for parasites, and present data expand the<br />
spectrum of fish hosts, to the effect that the list<br />
provides 75 new records for hosts and locations.<br />
Discussion<br />
Only 8 fish species have been examined in<br />
sufficient numbers to enable evaluation of helminth<br />
community composition and structure: Algansea<br />
lacustris, Chirostoma estor, C. attenu-<br />
Copyright © 2011, The Helminthological Society of Washington<br />
atutn, Goodea atripinnis, Alloophorus robustus,<br />
Allotoca diazi, Micropterus salmoides, and Cyprinus<br />
carpio. Patzcuaro Lake has been systematically<br />
sampled, while other localities have only<br />
been sampled occasionally and with few fish examined.<br />
From large areas of the basin no data<br />
on fish parasites exist at all. There is also limited<br />
information on the parasites of fish in rivers and<br />
other water bodies. Parasitological knowledge<br />
for the Lerma-Santiago river basin is fragmentary,<br />
as many studies did not record all the helminth<br />
species because they were prepared for<br />
taxonomic ends, such as the description of a single<br />
species. As a result, most of the research in<br />
the basin merely indicates where data are most<br />
needed.<br />
A notable aspect of the present data is a highly<br />
characteristic endemic helminth component in<br />
the Lerma-Santiago river basin. Of the 43 re-
SALGADO-MALDONADO ET AL.—HELMINTHS OF MEXICAN FISHES 207<br />
Table 2. Codes and features of the localities sampled or reported in the literature from which hosts were<br />
collected.<br />
Code<br />
Bata<br />
Bizn<br />
Chap<br />
Chic<br />
Coin<br />
Cons<br />
Cuit<br />
Igna<br />
Lagu<br />
Lerm<br />
Patz<br />
Rami<br />
Rsan<br />
Sala<br />
Taza<br />
Tila<br />
Trin<br />
Viet<br />
Zira<br />
Locality name<br />
Presa El Batan<br />
Presa La Biznaga<br />
Lago de Chapala<br />
Lago de Chicnahuapan<br />
("Almoloya del Rio")<br />
Presa Cointzio<br />
Presa Constitucion de 1917<br />
Lago de Cuitzeo<br />
Presa Ignacio Allende<br />
La Lagunilla<br />
Cienega de Lerma<br />
Lago de Patzcuaro<br />
Presa Ignacio Ramirez<br />
Rio Santiago (Aguamilpa)<br />
Lago de Salazar<br />
Las Tazas<br />
Santiago Tilapa, Laguna de<br />
Guadalupe Victoria<br />
Trinidad Fabela<br />
Villa Victoria<br />
Lago de Zirahuen<br />
Habitat<br />
type<br />
AR*<br />
AR<br />
NL<br />
NL<br />
AR<br />
AR<br />
NL<br />
AR<br />
WL<br />
WL<br />
NL<br />
AR<br />
RI<br />
NL<br />
AR<br />
NL<br />
AR<br />
AR<br />
NL<br />
<strong>State</strong> (coordinates)<br />
Queretaro (20°13'13"N; 100°24'39"W)<br />
Guanajuato (21°25'30"N; 100°52'52.7"W)<br />
Jalisco (20°08'-20°22'N; 102°42'-103°25'W)<br />
=|: AR = Artificial reservoir; NL = natural lake; WL = wetland; RI = river.<br />
corded helminth species, 6 (14%) are endemic<br />
to the basin: the digeneans A. mexicanum and<br />
M. bravoae, from atherinids and the goodeid G.<br />
multiradiatus, respectively; the monogenean O.<br />
mexicanum, a parasite of the cyprinid A. lacustris',<br />
and the nematodes R. lichtenfelsi, from the<br />
goodeids A. robustus, A. diazi, and G. atripinnis,<br />
and a species of Spinitectus, previously referred<br />
to as S. carolini, from the atherinids C. attenuatum<br />
and C. estor. Additionally, the nematode<br />
species B. nayaritensis, a parasite of C. beani in<br />
the Santiago River, may be endemic to this basin,<br />
because there is no other record of this species<br />
in Mexico (Moravec, 1998), and cichlids<br />
are the best studied fish family from a parasitological<br />
point of view (Salgado-Maldonado et<br />
al., 1997; Vidal-Martfnez and Kennedy, 2000).<br />
It is thought that the present hydrological configuration<br />
of the Lerma-Santiago river basin was<br />
created during the Pliocene Age by orogenic activity<br />
that isolated it from the ocean (Barbour,<br />
1973; Echelle and Echelle, 1984). The fish fauna<br />
of the basin consists of the descendants of marine<br />
ancestors that invaded the freshwater bodies,<br />
as well as Nearctic components such as cyprinids.<br />
It is assumed that by at least 5 million yr<br />
ago the fish species in the basin had established<br />
themselves, evolving and diversifying from their<br />
Estado de Mexico (19°11'N; 99°30'W)<br />
Michoacan (19°36'46"N; 101°17'58"W)<br />
Queretaro (20°25'00"N; 100°05'00"W)<br />
Guanajuato-Michoacan (20°04'34"- 1 9°53'25"N; 101° 1 9'34"-l 00°50'20"W)<br />
Guanajuato (20°55'N; 100°50'W)<br />
Estado de Mexico (19°08'30"N; 99°30'12"W)<br />
Estado de Mexico (19°22'41"N; 99°59'39"W)<br />
Michoacan (19°41'-19°32'N; 101°27'-101°53'W)<br />
Estado de Mexico (19°26'54"N; 99°59'32"W)<br />
Nayarit (21°46'42"N; 104°55'36"W)<br />
Estado de Mexico (19°21'5"N; 99°21'55"W)<br />
Estado de Mexico (not located)<br />
Estado de Mexico (19°1 1'15"N; 99°23'56"W)<br />
Estado de Mexico (19°48'N; 99°46'W)<br />
Estado de Mexico (19°26'28"N; 100°4'33"W)<br />
Michoacan (19°21'14"-19°29'32"N; 101°30'33"-101°46'15"W)<br />
original marine ancestors. The parasite fauna<br />
must also have evolved and diversified during<br />
this period of isolation, the current assemblage<br />
of endemic helminth species being the product<br />
of these evolutionary processes. To the extent to<br />
which the fish species adapted to these environments<br />
and speciated within them, so did their<br />
helminth communities, with some being lost and<br />
others developing in the new hosts. In other<br />
words, both the fish of the Lerma-Santiago river<br />
basin, and their parasites developed in isolation.<br />
The fish parasite fauna of this basin is also<br />
enriched through colonization by allogenic species<br />
transported by birds. As a result, the fish<br />
helminth communities in the basin have an<br />
abundant (14 of the total 43 species) component<br />
of allogenic species that mature in, and are<br />
transported by, birds: C. complanatum, Diplostomum<br />
sp., P. minimum, C. formosanus, L. intestinalis,<br />
C. cf. ralli, P. caballeroi, P. cf. urseus,<br />
P. cochlearii, V. campylancristrota, V. mutabilis,<br />
Eustrongylid.es sp., Contracaecum sp.,<br />
and P. brevis, most of which occur throughout<br />
the American continent or are cosmopolitan.<br />
Many factors may have favored this colonization.<br />
They include the small size of the fish in<br />
this basin, their gregarious habits, their shallow<br />
water habitat, their status in the food web, and<br />
Copyright © 2011, The Helminthological Society of Washington
Table 3. Parasite-host list of helminths collected from fish of the Lerma-Santiago river basin of west-c<br />
Number<br />
of<br />
hosts<br />
exam- Preva<br />
ined mean in<br />
Locality<br />
Host/infection site(s)*<br />
Parasite<br />
Adult Trematoda<br />
Family Allocreadiidae Stossich, 1903<br />
AUocreadium mexicanurn Osorio-Sarabia, Perez and Salgado-Maldonado,<br />
1986<br />
6%<br />
4%<br />
23%<br />
13%<br />
24%<br />
"Low<br />
40%<br />
216<br />
195<br />
48<br />
8<br />
209<br />
64<br />
5<br />
Patz<br />
Patz<br />
Rsan<br />
Tila<br />
Patz<br />
Lagu<br />
Viet<br />
C. estorll<br />
C. attenuatum/l<br />
A. crystallina/I<br />
C. riojaill<br />
M. salmoides/Pc, I<br />
G. midtiradiatusl\I<br />
Crepidostomum cooperi Hopkins, 1931<br />
Margotrema bravoae Lamothe-Argumedo, 1970<br />
9<br />
Chap<br />
/. dugesill<br />
Family Gorgoderidae (Looss, 1901)<br />
Phyllodistomum lacustris (Loewen, 1929)<br />
6%<br />
17<br />
Igna<br />
A. tincella/l<br />
Larval Trematoda<br />
Family Cryptogonimidae Ciurea, 1933<br />
Cryptogonimidae gen. sp.<br />
Family Proterodiplostomidae Dubois, 1936<br />
Proterodiplostomurn sp.<br />
6%<br />
67%<br />
45%<br />
20%<br />
15%<br />
33%<br />
3%<br />
20%<br />
61%<br />
30%<br />
3%<br />
28%<br />
36%<br />
17<br />
15<br />
94<br />
50<br />
75<br />
3<br />
31<br />
5<br />
18<br />
20<br />
29<br />
46<br />
14<br />
Igna<br />
Rami<br />
Chic<br />
Lagu<br />
Rami<br />
Sala<br />
Trin<br />
Viet<br />
Bizn<br />
Igna<br />
Trin<br />
Viet<br />
Rami<br />
A. tincella/Bc<br />
N. sallei/Bc<br />
G. multiradiatus/Bc<br />
/Be<br />
/Be<br />
/Be<br />
/Be<br />
/Be<br />
G. atripinnis/Bc<br />
/Be<br />
/Be<br />
C. humboIdtianum/L,<br />
C. riojai/Bc<br />
90%<br />
9<br />
9<br />
Family Clinostomidae Liihe, 1901<br />
Clinostomum complanatum (Rudolphi, 1814)<br />
30<br />
41<br />
31<br />
Cuit<br />
Patz<br />
Patz<br />
A. robustusll<br />
/L, M<br />
A. diazi/L, M<br />
Copyright © 2011, The Helminthological Society of Washington
Table 3. Continued.<br />
Number<br />
of<br />
hosts<br />
exam- Preva<br />
ined mean i<br />
Locality<br />
Host/infection site(s)*<br />
Parasite<br />
13%<br />
0.6%<br />
5%<br />
27%<br />
30<br />
178<br />
22<br />
41<br />
Cuit<br />
Patz<br />
Igna<br />
Cuit<br />
G. atripinnisll<br />
/L<br />
/L<br />
X. variatusH<br />
Family Diplostomidae Poirier, 1886<br />
Diplostomum sp.<br />
10%<br />
79%<br />
9%<br />
11%<br />
7%<br />
14%<br />
30%<br />
5%<br />
216<br />
38<br />
22<br />
9<br />
30<br />
42<br />
30<br />
390<br />
6<br />
30<br />
41<br />
31<br />
9<br />
30<br />
178<br />
35<br />
41<br />
30<br />
195<br />
42<br />
216<br />
30<br />
30<br />
17<br />
15<br />
1<br />
8<br />
10<br />
118<br />
13<br />
Patz<br />
Bizn<br />
Rsan<br />
Igna<br />
Cuit<br />
Zira<br />
Cuit<br />
Patz<br />
Lerm<br />
Cuit<br />
Patz<br />
Patz<br />
Lerm<br />
Cuit<br />
Patz<br />
Patz<br />
Cuit<br />
Patz<br />
Patz<br />
Zira<br />
Patz<br />
Cuit<br />
Cuit<br />
Igna<br />
Rami<br />
Lagu<br />
Chic<br />
Igna<br />
Chic<br />
Rami<br />
C. estor/B<br />
C. jor'danifM.<br />
P. sphenops/Bc<br />
Y. alta/M<br />
G. atripinnisll<br />
C. attenuatum/1<br />
C. jordanill<br />
A. lacustris/M<br />
N. sallei/1<br />
A. robustusll<br />
/L, M, Mu, E<br />
A. diazi/L, M, Mu, E<br />
G. multiradiatusH<br />
G. atripinnisll<br />
/L, Mu<br />
/L, M, Mu, E<br />
X. variants!!<br />
C. attenuatum/1^, M, Mu<br />
/L, M, Mu, E, B<br />
/L, M, Mu<br />
C. estor/L, Mu, E, B<br />
C. jordani/1<br />
O. aureusll<br />
A. tincella/M<br />
N. sallei/M<br />
/M<br />
/M<br />
Y. altall, L, M<br />
G. multiradiatus/M<br />
/M<br />
Diplostomum (Tylodelphys) sp.<br />
Posthodiplostomum minimum (MacCallum, 1921) Dubois,<br />
1936<br />
93%<br />
87%<br />
62%<br />
80%<br />
100%<br />
98%<br />
81%<br />
95%<br />
67%<br />
7%<br />
82%<br />
7%<br />
100%<br />
22%<br />
80%<br />
22%<br />
8%<br />
Copyright © 2011, The Helminthological Society of Washington
Table 3. Continued.<br />
Preval<br />
mean in<br />
Number<br />
of<br />
hosts<br />
examined<br />
Locality<br />
Host/infection site(s)*<br />
Parasite<br />
60%<br />
55%<br />
25%<br />
57%<br />
5%<br />
100%<br />
100%<br />
48%<br />
52%<br />
100%<br />
26%<br />
8%<br />
25<br />
22<br />
4<br />
35<br />
22<br />
9<br />
2<br />
46<br />
23<br />
2<br />
23<br />
13<br />
Bizn<br />
Igna<br />
Trin<br />
Igna<br />
Rsan<br />
Igna<br />
Rami<br />
Viet<br />
Igna<br />
Igna<br />
Rami<br />
Tila<br />
G. atripinnis/M<br />
/L, M<br />
/M<br />
X. variatusfL, M<br />
P. sphenops/M<br />
P. infans/L, M<br />
/M<br />
C. humboldtianumfL<br />
C. jordani/L, M<br />
C. labarcaefL, M<br />
C. riojaifL, M<br />
/L, M<br />
?<br />
31<br />
Patz<br />
A. diazill<br />
Family Plagiorchiidae Liihe, 1901<br />
Ochetosorna sp.<br />
Family Heterophyidae Odhner, 1914<br />
Centrocestus formosanus (Nishigori, 1924)<br />
18%<br />
50%<br />
27%<br />
100%<br />
20%<br />
62%<br />
38%<br />
50%<br />
17<br />
14<br />
11<br />
1<br />
5<br />
13<br />
48<br />
2<br />
Igna<br />
Igna<br />
Igna<br />
Rsan<br />
Igna<br />
Rsan<br />
Rsan<br />
Rsan<br />
A. tincella/G<br />
Y. alta/G<br />
G. atripinnis/G<br />
P. sphenops/G<br />
P. infans/G<br />
Poeciliopsis sp./G<br />
A. crystallina/G<br />
L. macrochirus/G<br />
L2%<br />
25<br />
Rsan<br />
C. beani/G<br />
Monogenea<br />
Family Dactylogyridae Bychowsky, 1933<br />
Sciadicleithrum sp.<br />
28%<br />
23%<br />
46<br />
22<br />
Chic<br />
Rsan<br />
G. multiradiatuslG<br />
P. sphenopslG<br />
Family Gyrodactylidae Cobbold, 1864<br />
Gyrodactylus elegans Nordmann, 1832<br />
Gyrodactyhis sp.<br />
62%<br />
390<br />
Patz<br />
A. lacustrislG<br />
Family Discocotylidae Price, 1936<br />
Octomacrum mexicanum Lamothe-Argumedo, 1981<br />
Copyright © 2011, The Helminthological Society of Washington
Table 3. Continued.<br />
Number<br />
of<br />
hosts<br />
exam- Preva<br />
ined mean i<br />
Locality<br />
Host/infection site(s)*<br />
Parasite<br />
0.3%<br />
390<br />
Patz<br />
A. lacustns/l<br />
Adult Cestoda<br />
Order Caryophyllidea van Beneden in Carus, 1863<br />
Caryophyllidea gen. sp.<br />
9<br />
Family Bothriocephalidae Blanchard, 1849<br />
Bothriocephalus acheilognathi Yamaguti, 1934<br />
5%<br />
13%<br />
390<br />
6<br />
178<br />
5<br />
41<br />
31<br />
9<br />
41<br />
9<br />
36<br />
30<br />
195<br />
42<br />
216<br />
234<br />
9<br />
9<br />
25<br />
43<br />
209<br />
40<br />
17<br />
15<br />
17<br />
3<br />
43<br />
2<br />
63<br />
50<br />
75<br />
Chap<br />
Patz<br />
Lerm<br />
Patz<br />
Lerm<br />
Patz<br />
Patz<br />
Lerm<br />
Bata<br />
Chap<br />
Cons<br />
Patz<br />
Patz<br />
Zira<br />
Patz<br />
Coin<br />
Chap<br />
Patz<br />
Bata<br />
Cons<br />
Patz<br />
Cons<br />
Igna<br />
Rami<br />
Igna<br />
Rami<br />
Rami<br />
Trin<br />
Chic<br />
Lagu<br />
Rami<br />
A. mbescensll<br />
A. lacustrisll<br />
N. sallei/I<br />
C. carpioll<br />
l\ robustus/l<br />
12%<br />
8%<br />
13%<br />
7%<br />
24%<br />
2%<br />
2%<br />
24%<br />
40%<br />
l%<br />
8%<br />
6%<br />
13%<br />
24%<br />
33%<br />
2%<br />
100%<br />
3%<br />
26%<br />
3%<br />
A. diazifl<br />
G. multiradiatusll<br />
G. atripinnisll<br />
/I<br />
X. variatusll<br />
C. attenuatum/l<br />
/I<br />
/I<br />
C. estor/l<br />
C. humboldtianumll<br />
C. ocotlanell<br />
C. grandocule/l<br />
Cliirostoma sp./I<br />
/I<br />
M. salmoidesll<br />
O. niloticiis/l<br />
A. tincella/l<br />
N. sallei/l<br />
Y. alta/l<br />
/I<br />
C. carpioll<br />
11<br />
G. miiltiracliatus/l<br />
l\<br />
Copyright © 2011, The Helminthological Society of Washington
212 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
_| H * < -l<br />
O O O O O O<br />
=: £ £ 5: =: 5<br />
a! a
Table 3. Continued.<br />
Number<br />
of<br />
hosts<br />
exam- Prev<br />
ined mean<br />
Locality<br />
Host/infection site(s)*<br />
Parasite<br />
8<br />
3%<br />
22%<br />
3<br />
2%<br />
2%<br />
33%<br />
10<br />
4%<br />
9%<br />
3%<br />
4%<br />
24<br />
38<br />
23<br />
38<br />
48<br />
46<br />
3<br />
20<br />
50<br />
75<br />
31<br />
25<br />
Igan<br />
Bizn<br />
Igna<br />
Bizn<br />
Rsan<br />
Viet<br />
Rami<br />
Rami<br />
Lagu<br />
Rami<br />
Trin<br />
Rsan<br />
X. variatus/M<br />
C. jordani/M, L<br />
C. jordani/L<br />
C. jordanifL<br />
A. crystallinafL<br />
C. humboldtianum/Gb<br />
C. jordani/Gb<br />
C. riojai/Gb<br />
G. miiltiradiatus/Gb<br />
Paradilepis caballeroi Rysavy and Macko, 1973<br />
Paradilepis cf. urceus (Wedl, 1855)<br />
Paradilepis sp.<br />
Parvitaenia cochlearii Coil, 1955<br />
Valipora campy lancristrota (Wedl, 1855)<br />
C. beani/L, Gb<br />
Valipora mutabilis Linton, 1927<br />
0.5%<br />
195<br />
Patz<br />
C. attenuatum/l<br />
Order Cyclophyllidea<br />
Cyclophyllidea gen. sp.<br />
Adult Nematoda<br />
Family Capillariidae Neveau-Lemaire, 1936<br />
Pseudocapillaria tomentosa (Dujardin, 1843)<br />
5%<br />
10<br />
0.5%<br />
2%<br />
1<br />
7%<br />
8%<br />
5%<br />
33%<br />
20<br />
178<br />
195<br />
216<br />
110<br />
43<br />
75<br />
184<br />
3<br />
Patz<br />
Patz<br />
Patz<br />
Patz<br />
Patz<br />
Patz<br />
Patz<br />
Patz<br />
Rami<br />
A. robustiis/l<br />
G. atripinnis/I<br />
C. attenuatum/I<br />
C. estorfl<br />
/I<br />
C. carpio/l<br />
N. sallei/l<br />
(1986) but Moravec et al<br />
5%<br />
4%<br />
20<br />
25<br />
Igna<br />
Bizn<br />
Remarks: This species was originally described as Capillaria patzcuarensis by Osorio-Sarabia et al.<br />
from Europe with cyprinids.<br />
Capillariidae gen. sp.<br />
G. atripinnis/I<br />
/I<br />
14%<br />
7<br />
Rsan<br />
C. beani/I<br />
Family Cucullanidae Cobbold, 1864<br />
Dichelyne mexicanus Caspeta-Mandujano, Moravec and<br />
Salgado-Maldonado, 1999<br />
Copyright © 2011, The Helminthological Society of Washington
Table 3. Continued.<br />
Number<br />
of<br />
hosts<br />
exam- Preva<br />
ined mean in<br />
* Locality<br />
Parasite Host/infection site(s)<br />
0.5%<br />
390<br />
Patz<br />
Family Philometridae Baylis and Daubney, 1926<br />
Philometridae gen. sp. A. lacustrisfBc<br />
7<br />
7<br />
7<br />
40%<br />
8%<br />
7<br />
44%<br />
360<br />
41<br />
31<br />
20<br />
178<br />
35<br />
25<br />
Cuit<br />
Patz<br />
Patz<br />
Cuit<br />
Patz<br />
Patz<br />
Rsan<br />
Family Rhabdochonidae Travassos, Artigas, and Pereira, 1928<br />
Rhabdochona lichtenfelsi Sanchez-Alvarez, Garcia, and A. robustusfl<br />
Perez, 1998 /I<br />
A. diazi/i<br />
G. atripinnis/l<br />
n<br />
n<br />
Beaninema nayaritense Caspeta-Mandujano, Moravec, and C. beani/l<br />
Salgado-Maldonado, 2000<br />
10%<br />
14%<br />
43%<br />
12%<br />
30<br />
195<br />
42<br />
216<br />
Patz<br />
Patz<br />
Zira<br />
Patz<br />
Family Cystidicolidae Skrjabin, 1846<br />
Spinitectus sp. C. attenuatum/l<br />
11<br />
l\ estor/I<br />
Remarks: The nematodes were originally reported as Spinitectus carolini Holl, 1928, but they in fact belong to a separate spec<br />
of the Czech Republic, personal communication).<br />
Larval Nematodes<br />
Family Dioctophymatidae Railliet, 1915<br />
?<br />
2%<br />
2%<br />
13%<br />
1%<br />
10%<br />
41<br />
178<br />
195<br />
30<br />
209<br />
10<br />
Patz<br />
Patz<br />
Patz<br />
Patz<br />
Patz<br />
Bizn<br />
A. robustus/M, Be<br />
G. atripinnis/Mu<br />
C. attenuatum/M<br />
n<br />
M. salmoidesfMu<br />
G. atripinnis/Mu<br />
Eustrongylides sp.<br />
Family Anisakidae Railliet and Henry, 1912<br />
Contracaeciim sp.<br />
0.3%<br />
6%<br />
13%<br />
1 1 %<br />
5%<br />
31%<br />
390<br />
17<br />
15<br />
9<br />
22<br />
35<br />
Patz<br />
Igna<br />
Rami<br />
Igna<br />
Igna<br />
Igna<br />
A. lacustris/l<br />
A. tincella/M<br />
N. sallei/M<br />
Y. alta /M<br />
G. atripinnis/M, Be<br />
X. variatus/M<br />
Copyright © 2011, The Helminthological Society of Washington<br />
COMPARA1
Table 3. Continued.<br />
Number<br />
of<br />
hosts<br />
exam- Prev<br />
ined mean<br />
Locality<br />
Host/infection site(s)*<br />
Parasite<br />
119<br />
3<br />
17<br />
12<br />
50%<br />
9<br />
38<br />
23<br />
25<br />
2<br />
Igna<br />
Bizn<br />
Igna<br />
Rsan<br />
Rsan<br />
P. infansIM<br />
C. joi'dani/M<br />
/M<br />
C. beani/L,, M<br />
A. tajasica/M<br />
5%<br />
0.8%<br />
2%<br />
Patz<br />
Patz<br />
Patz<br />
Patz<br />
Patz<br />
Patz<br />
Patz<br />
Patz<br />
Rami<br />
Rami<br />
Bizn<br />
Trin<br />
Igna<br />
A. robustiisfL<br />
A. laciistris/l<br />
C. carpioll<br />
A. ?-obnstns/M, I<br />
A. diazi/M, I<br />
G. atripinnis/M, I<br />
/I<br />
M. salmoidesfl<br />
N. salleifl<br />
G. miiltiradiatiis/I<br />
G. atripinnisfl<br />
fl<br />
X. variatus/l<br />
Family Gnathostomatidae Railliet, 1895<br />
Gnathostoma sp.<br />
Spiroxys sp.<br />
1<br />
l%<br />
33<br />
15<br />
6<br />
3<br />
5<br />
20<br />
390<br />
184<br />
41<br />
31<br />
35<br />
178<br />
209<br />
3<br />
13<br />
18<br />
29<br />
21<br />
20%<br />
25<br />
Rsan<br />
C. beanifl<br />
Acanthocephala Adult<br />
Family Neoechinorhynchidae Ward, 1953<br />
Neoechinorhynchus golvani Salgado-Maldonado, 1978<br />
Acanthocephala Larvae<br />
Family Polymorphidae Meyer, 1931<br />
Polymorphic brevis Van Cleave, 1916<br />
0.3%<br />
2%<br />
3%<br />
4%<br />
8%<br />
3%<br />
6%<br />
390<br />
184<br />
41<br />
31<br />
178<br />
195<br />
216<br />
209<br />
35<br />
Patz<br />
Patz<br />
Patz<br />
Patz<br />
Patz<br />
Patz<br />
Patz<br />
Patz<br />
Igna<br />
A. lacustris/M<br />
C. carpio/L., M<br />
A. robustus/M, Mu<br />
A. diazi/M, Mu<br />
G. atripinnis/L,, M<br />
C. attenuatum/L., M<br />
C. estor/L, M<br />
M. salmoides/L,, M<br />
X. variatus/L,<br />
* B = brain; Be = body cavity; E = eyes; G = gills; Gb = gall bladder; I = intestine; L = liver; M = mesentery; Mu =<br />
Copyright © 2011, The Helminthological Society of Washington
216 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
their situation along the annual migratory routes<br />
of Nearctic birds. Additionally, the low number<br />
of helminth species in the basin may have readily<br />
allowed invasion of these communities by<br />
allogenic species.<br />
Three helminth species on the list are recent,<br />
anthropogenically introduced colonizers. The<br />
first is the cestode B. acheilognathi, which is the<br />
most widely distributed species in the basin and<br />
is found in 22 host species. The second is the<br />
heterophyid trematode C. formosanus. The actual<br />
distribution of this helminth within the basin<br />
has not been evaluated, because the intermediate<br />
host, the thiarid snail Melanoides tuberculata<br />
Miiller, 1774, has established along riverbanks<br />
and in riverbeds, where few fish have been sampled.<br />
Both these species were introduced recently<br />
into Mexico; the cestode together with<br />
Asian carp (Salgado-Maldonado et al., 1986),<br />
and the trematode most probably with the intermediate<br />
snail host (Scholz and Salgado-Maldonado,<br />
2000). The third species is the capillariid<br />
nematode P. tomentosa, reported from atherinids<br />
and goodeids, as well as from cultured carp, C.<br />
carpio, from Mexico, where it was probably introduced<br />
along with its fish host from Europe<br />
(Moravec, 1998; Moravec et al., <strong>2001</strong>).<br />
The proportions among the helminth groups<br />
that constitute the communities in the fish of the<br />
Lerma-Santiago river basin are also distinctive.<br />
The dominance in species numbers of nematodes<br />
and trematodes (principally metacercariae)<br />
is a pattern characteristic of the fish helminth<br />
communities of southeastern Mexico (Scholz et<br />
al., 1995; Salgado-Maldonado and Kennedy,<br />
1997; Scholz and Vargas-Vazquez, 1998) and<br />
the Balsas River basin in central Mexico (Salgado-Maldonado<br />
et al., <strong>2001</strong>). However, data in<br />
the present study show that cestodes, both adults<br />
and metacestodes, are almost as important in the<br />
Lerma-Santiago river basin in terms of numbers<br />
as the nematodes and trematodes. Most cestodes<br />
found, such as V. campylancristrota, occur<br />
throughout the American continent or are cosmopolitan<br />
(see Scholz and Salgado-Maldonado,<br />
<strong>2001</strong>). The presence of 5 monogenean species<br />
in the basin is also notable, as it is a higher<br />
number than recorded in other drainages in central<br />
Mexico. However, the monogenean fauna of<br />
freshwater fishes in southeastern Mexico is even<br />
richer (Kritsky et al., 1994, 2000; Mendoza-<br />
Franco et al., 1997, 1999, 2000).<br />
It is still not possible to form conclusions<br />
Copyright © 2011, The Helminthological Society of Washington<br />
about the zoogeographic characteristics of the<br />
fish helminth parasite communities in the Lerma-Santiago<br />
river basin, as very few studies<br />
have been done. However, the data that do exist<br />
suggest that the proportion of endemic parasites<br />
is high, and thus very distinctive, as compared<br />
for example to the lack of endemic species<br />
among the helminth parasites of fishes from the<br />
Balsas River drainage (Salgado-Maldonado et<br />
al., <strong>2001</strong>). The helminth communities were<br />
probably initially poor, and have been invaded<br />
by allogenic, Nearctic species transported by<br />
birds that have enriched these multispecific assemblages.<br />
Research into fish helminth parasites in the<br />
Lerma-Santiago river basin has been restricted to<br />
descriptions of some species, and more detailed<br />
studies have been carried out only in Patzcuaro<br />
Lake. Obviously, more complete inventories of<br />
the fish parasites in this basin are urgently required.<br />
Almost 7% of the fish species that originally<br />
inhabited the basin are extinct, and an additional<br />
23% are classified as endangered or vulnerable<br />
because of population decline associated<br />
with continuous habitat degradation and introduction<br />
of competing and predatory species that are<br />
added to the natural predation pressures in these<br />
ecosystems (Soto-Galera et al., 1998).<br />
Acknowledgments<br />
This study was supported by project no.<br />
276<strong>68</strong>N from the Consejo Nacional de Ciencia<br />
y Tecnologia (CONACyT), Mexico, and by project<br />
no. H007 from the Comision Nacional para<br />
el Estudio y Uso de la Biodiversidad (CONA-<br />
BIO), Mexico. We are indebted to Dr. Frantisek<br />
Moravec for confirmation of identification of<br />
nematodes and Dr. Tomas Scholz for identification<br />
of cestodes. We also thank Nancy Minerva<br />
Lopez-Flores, Isabel Jimenez-Garcia, Cris Caneda-Guzman,<br />
Rafael Baez-Vale, Norman Mercado-Silva,<br />
and Felipe Villegas-Marquez for<br />
their assistance in the field and laboratory.<br />
Literature Cited<br />
Alarcon, G. C. 1988. Diagnostico e identificacion de<br />
una parasitosis helmmtica en Carassius carassius<br />
en un centre piscicola. Revista Latinoamericana<br />
de Microbiologia 30:297.<br />
, and J. L. Castro-Aguirre. 1988. Tratamiento<br />
experimental con Mebendazol para bothriocefalosis<br />
en Carassius carassius. Revista Latinoamericana<br />
de Microbiologfa 30:298.<br />
Astudillo-Ramos, L., and E. Soto-Galera. 1997. Es-
tudio helmintologico de Chirostoma humboldtianum<br />
y Girardinichthys multiradiatus capturados en<br />
el Lerma. Zoologia Informa 35:53-59.<br />
Barbour, C. D. 1973. A biogeographical history of<br />
Chirostoma (Pisces: Atherinidae): a species flock<br />
from the Mexican Plateau. Copeia 1973:533-556.<br />
Caspeta-Mandujano, J. M., F. Moravec, and G. Salgado-Maldonado.<br />
1999. Observations on cucullanid<br />
nematodes from freshwater fishes in Mexico,<br />
including Dichelyne mexicanus sp. n. Folia Parasitologica<br />
46:289-295.<br />
, , and . <strong>2001</strong>. Two new species<br />
of rhabdochonids (Nematoda: Rhabdochonidae)<br />
from freshwater fishes in Mexico, with a description<br />
of a new genus. Journal of <strong>Parasitology</strong> 87:<br />
139-143.<br />
Diaz-Pardo, E., M. A. Godinez-Rodriguez, E. Lopez-Lopez,<br />
and E. Soto-Galera. 1993. Ecologia<br />
de los peces de la cuenca del rio Lerma, Mexico.<br />
Anales de la Escuela Nacional de Ciencias Biologicas<br />
39:103-127.<br />
Echelle, A. A., and A. F. Echelle. 1984. Evolutionary<br />
genetics of a "species flock": atherinid fishes on<br />
the Mesa Central of Mexico. Pages 93-110 in A.<br />
A. Echelle and I. Kornfield, eds. Evolution of Fish<br />
Species Flocks. University of Maine at Orono<br />
Press, Orono, Maine, U.S.A.<br />
Espinosa-Huerta, E., L. Garcia-Prieto, and G.<br />
Perez. 1996. Helminth community structure of<br />
Chirostoma attemiatiim (Osteichthyes: Atherinidae)<br />
in two Mexican lakes. Southwestern Naturalist<br />
41:288-292.<br />
Espinosa-Perez, H. 1993. Riqueza y diversidad de peces.<br />
Ciencias, Mexico 7:77-84.<br />
Flores-Barroeta, L. 1953. Cestodos de vertebrados I.<br />
Ciencia 13:31-36.<br />
Garcia-Prieto, L., H. Mejia, and G. Perez. 1988.<br />
Hallazgo del plerocercoide de Ligula intestinalis<br />
(Cestoda) en algunos peces dulceacuicolas de<br />
Mexico. Anales del Institute de Biologia, Universidad<br />
Nacional Autonoma de Mexico, Serie<br />
Zoologia 58:887-888.<br />
, and D. Osorio-Sarabia. 1991. Distribucion<br />
actual de Bothriocephalus acheilognathi en Mexico.<br />
Anales del Institute de Biologia, Universidad<br />
Nacional Autonoma de Mexico, Serie Zoologia<br />
62:523-526.<br />
Guzman-Cornejo, M. C., and L. Garcia-Prieto.<br />
1999. Trematodiasis en algunos peces del lago de<br />
Cuitzeo, Michoacan, Mexico. Revista de Biologia<br />
Tropical 47:593=596.<br />
Kritsky, D. C., E. F. Mendoza-Franco, and T.<br />
Scholz. 2000. Neotropical Monogenoidea. 36.<br />
Dactylogyrids from the gills of Rhamdia guatemalcnsis<br />
(Siluriformes: Pimelodidae) from cenotes<br />
of the Yucatan Peninsula, Mexico, with proposal<br />
of Ameloblastella gen. n. and Aphanoblastella<br />
gen. n. (Dactylogyridae: Ancyrocephalinae).<br />
<strong>Comparative</strong> <strong>Parasitology</strong> 67:76-84.<br />
, V. M. Vidal-Martinez, and R. Rodriguez-<br />
Canul. 1994. Neotropical Monogenoidea 19. Dactylogyridae<br />
of cichlids (Perciformes) from the Yucatan<br />
Peninsula, with descriptions of three new<br />
species of Sciadicleithrum Kritsky, Thatcher, and<br />
SALGADO-MALDONADO ET AL.—HELMINTHS OF MEXICAN FISHES 217<br />
Boeger, 1989. Journal of the Helminthological Society<br />
of Washington 61:26-33.<br />
Lamothe-Argumedo, R. 1970. Trematodos de peces<br />
VI. Margotrema bravoae gen. nov. sp. nov.<br />
(Trematoda: Allocreadiidae) parasito de Lermichthys<br />
multiradiatus Meek. Anales del Institute de<br />
Biologia, Universidad Nacional Autonoma de<br />
Mexico, Serie Zoologia 41:87-92.<br />
. 1981. Monogeneos parasitos de peces VIII.<br />
Descripcion de una nueva especie del genero Octomacrum<br />
Miiller, 1934 (Monogenea: Discocotylidae).<br />
Anales del Institute de Biologia, Universidad<br />
Nacional Autonoma de Mexico, Serie Zoologia<br />
51:56-60.<br />
. 1988. Trematodos de peces VIII. Primer registro<br />
de Phyllodistomum lacustri (Loewen, 1924)<br />
parasito de Ictalurus dugesi en Mexico. Anales<br />
del Institute de Biologia, Universidad Nacional<br />
Autonoma de Mexico, Serie Zoologia 58:487-<br />
496.<br />
-, and A. Cruz-Reyes. 1972. Hallazgo de Ligula<br />
intestinalis (Goeze, 1782) Gmelin, 1790 en<br />
Lermichthys multiradiatus (Meek) (Pisces: Goodeidae).<br />
Revista de la Sociedad Mexicana de Historia<br />
Natural 33:99-100.<br />
Leon-Regagnon, V. 1992. Fauna helmintologica de<br />
algunos vertebrados acuaticos de la Cienega de<br />
Lerma, Mexico. Anales del Institute de Biologia,<br />
Universidad Nacional Autonoma de Mexico, Serie<br />
Zoologia 63:151-153.<br />
Margolis, L., G. W. Esch, J. C. Holmes, A. M. Kuris,<br />
and G. A. Schad. 1982. The use of ecological<br />
terms in parasitology (report of an ad hoc committee<br />
of the American Society of Parasitologists).<br />
Journal of <strong>Parasitology</strong> <strong>68</strong>:131-133.<br />
Mendoza-Franco, E. F., T. Scholz, and V. M. Vidal-<br />
Martinez. 1997. Sciadicleithrum meekii sp. n.<br />
(Monogenea: Ancyrocephalinae) from the gills of<br />
Cichlasoma meeki (Pisces: Cichlidae) from cenotes<br />
( = sinkholes) of the Yucatan Peninsula, Mexico.<br />
Folia Parasitologica 44:205-208.<br />
, , C. Vivas-Rodriguez, and J. Vargas-<br />
Vazquez. 1999. Monogeneans of freshwater fishes<br />
from cenotes (sinkholes) of the Yucatan Peninsula,<br />
Mexico. Folia Parasitologica 46:267-273.<br />
-, V. M. Vidal-Martinez, M. L. Aguirre-Macedo,<br />
R. Rodriguez-Canul, and T. Scholz. 2000.<br />
Species of Sciadicleithrum (Dactylogyridae: Ancyrocephalinae)<br />
of cichlid fishes from southeastern<br />
Mexico and Guatemala: new morphological<br />
data and host and geographical records. <strong>Comparative</strong><br />
<strong>Parasitology</strong> 67:85-91.<br />
Mendoza-Garfias, B., L. Garcia-Prieto, and G.<br />
Perez. 1996. Helmintos de la "acumara" Algansea<br />
lacustris en el lago de Patzcuaro, Michoacan,<br />
Mexico. Anales del Instituto de Biologia, Universidad<br />
Nacional Autonoma de Mexico, Serie<br />
Zoologia 67:77=88.<br />
Miller, R. R. 1982. Pisces. Pages 486-581 in S. H.<br />
Hurlbert and A. Villalobos, eds. Aquatic Biota of<br />
Mexico, Central America and the West Indies. San<br />
Diego <strong>State</strong> University, San Diego, California,<br />
U.S.A.<br />
Moravec, F. 1998. Nematodes of Freshwater Fishes<br />
Copyright © 2011, The Helminthological Society of Washington
218 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
of the Neotropical Region. Academy of Sciences<br />
of the Czech Republic, Prague, Czech Republic.<br />
464 pp.<br />
, R. Aguilar-Aguilar, and G. Salgado-Maldonado.<br />
<strong>2001</strong>. Systematic status of Capillaria<br />
patzcuarensis Osorio-Sarabia, Perez-Ponce de<br />
Leon et Salgado-Maldonado, 1986 (Nematoda:<br />
Capillariidae) from freshwater fishes in Mexico.<br />
Acta Parasitologica 46:8-11.<br />
-, G. Salgado-Maldonado, and D. Osorio-Sarabia.<br />
2000. Records of the bird capillariid, Ornithocapillaria<br />
appendiculata (Freitas, 1933)<br />
comb, n., from freshwater fishes in Mexico, with<br />
remarks on Capillaria patzcuarensis Osorio-Sarabia,<br />
Perez-Ponce de Leon and Salgado-Maldonado,<br />
1986. Systematic <strong>Parasitology</strong> 45:53-59.<br />
Osorio-Sarabia, D., G. Perez, and G. Salgado-Maldonado.<br />
1986. Helmintos de peces del lago de<br />
Patzcuaro, Michoacan, I. Helmintos de Chirostoma<br />
estorel "pescado bianco." Taxonomfa. Anales<br />
del Instituto de Biologfa, Universidad Nacional<br />
Autonoma de Mexico, Serie Zoologfa 57:61-92.<br />
Peresbarbosa-Rojas, R. E., G. Perez, and L. Garcia.<br />
1994. Helmintos parasites de tres especies de peces<br />
(Goodeidae) del lago de Patzcuaro, Michoacan.<br />
Anales del Instituto de Biologia, Universidad<br />
Nacional Autonoma de Mexico, Serie Zoologfa<br />
65:201-204.<br />
Perez-Ponce de Leon, G., B. Mendoza G., and G.<br />
Pulido F. 1994. Helminths of the charal prieto<br />
Chirostoma atteniiatum (Osteichthes: Atherinidae),<br />
from Patzcuaro Lake, Michoacan, Mexico.<br />
Journal of the Helminthological Society of Washington<br />
61:139-141.<br />
Pineda-Lopez, R., and C. Gonzalez. 1997. Bothriocephalus<br />
acheilognathr. presencia e importancia<br />
de un invasor asiatico infectando peces de Queretaro.<br />
Zoologfa Informa 35:5-12.<br />
Salgado-Maldonado, G., G. Cabanas-Carranza, J.<br />
M. Caspeta-Mandujano, E. Soto-Galera, E.<br />
Mayen-Pena, D. Brailovsky, and R. Baez-Vale.<br />
<strong>2001</strong>. Helminth parasites of freshwater fishes of<br />
the Balsas River drainage, southwestern Mexico.<br />
<strong>Comparative</strong> <strong>Parasitology</strong> <strong>68</strong>:196-203.<br />
, S. Guillen-Hernandez, and D. Osorio-Sarabia.<br />
1986. Presencia de Bothriocephalus acheilognathi<br />
Yamaguti, 1934 (Cestoda: Bothriocephalidae)<br />
en peces de Patzcuaro, Michoacan, Mexico.<br />
Anales del Instituto de Biologfa, Universidad Nacional<br />
Autonoma de Mexico, Serie Zoologfa 57:<br />
213-218.<br />
Copyright © 2011, The Helminthological Society of Washington<br />
, and C. R. Kennedy. 1997. Richness and similarity<br />
of helminth communities in the tropical<br />
cichlid fish Cichlasoma iirophthalmus from the<br />
Yucatan Peninsula, Mexico. <strong>Parasitology</strong> 1 14:<br />
581-590.<br />
, and D. Osorio-Sarabia. 1987. Helmintos de<br />
algunos peces del lago de Patzcuaro. Ciencia y<br />
Desarrollo 113:41-57.<br />
-, R. Pineda-Lopez, V. M. Vidal-Martinez,<br />
and C. R. Kennedy. 1997. A checklist of mctazoan<br />
parasites of cichlid fish from Mexico. Journal<br />
of the Helminthological Society of Washington<br />
64:195-207.<br />
Sanchez-Alvarez, A., L. Garcia-Prieto, and G.<br />
Perez. 1998. A new species of Rhahdochona Railliet,<br />
1916 (Nematoda: Rhabdochonidae) from endemic<br />
goodeids (Cyprinodontiformes) from two<br />
Mexican lakes. Journal of <strong>Parasitology</strong> 84:840-<br />
845.<br />
Scholz, T., and G. Salgado-Maldonado. 2000. The<br />
introduction and dispersal of Centrocestus forinosanus<br />
(Nishigori, 1924) (Digenea: Heterophyidae)<br />
in Mexico: a review. American Midland Naturalist<br />
143:185-200.<br />
, and . <strong>2001</strong>. Metacestodes of the family<br />
Dilepididae (Cestoda: Cyclophyllidea) parasitizing<br />
fishes in Mexico. Systematic <strong>Parasitology</strong><br />
49:23-39.<br />
, and J. Vargas-Vazquez. 1998. Trematodes<br />
from fishes of the Rio Hondo River and freshwater<br />
lakes of Quintana Roo, Mexico. Journal of the<br />
Helminthological Society of Washington 65:91-<br />
95.<br />
-, F. Moravec, C. Vivas-Rodriguez,<br />
and E. Mendoza-Franco. 1995. Metacercariae of<br />
trematodes of fishes from cenotes ( = sinkholes) of<br />
the Yucatan Peninsula, Mexico. Folia Parasitologica<br />
42:173-192.<br />
Soto-Galera, E., E. Diaz-Pardo, E. Lopez-Lopez,<br />
and J. Lyons. 1998. Fish as indicators of environmental<br />
quality in the Rio Lerma Basin, Mexico.<br />
Aquatic Ecosystem Health and Management<br />
1:267-216.<br />
Vidal-Martinez, V. M., and C. R. Kennedy. 2000.<br />
Zoogeographical determinants of the helminth<br />
fauna composition of Neotropical cichlid fish.<br />
Pages 250-278 in G. Salgado-Maldonado, A. N.<br />
Garcfa-Aldrete, and V. M. Vidal-Martfnez, eds.<br />
Metazoan Parasites in the Neotropics: A Systematic<br />
and Ecological Perspective. Instituto de Biologfa,<br />
Universidad Nacional Autonoma de Mexico,<br />
Mexico City, Mexico.
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 219-227<br />
Singhiatrema vietnamensis sp. n. (Digenea: Ommatobrephidae) and<br />
Szidatia taiwanensis (Fischthal and Kimtz, 1975) comb. n. (Digenea:<br />
Cyathocotylidae) from Colubrid Snakes in Vietnam<br />
STEPHEN S. CURRAN,' ROBIN M. OVERSTREET,U DANG TAT THE,2 AND NGUYEN THI LE2<br />
1 The University of Southern Mississippi, Gulf Coast Research Laboratory, P.O. Box 7000, Ocean Springs,<br />
Mississippi 39566, U.S.A. (e-mail: robin.overstreet@usm.edu and stephen.curran@usm.edu) and<br />
2 Institute of Ecology and Biological Resources, Nghia do, Cau Giay, Hanoi, Vietnam (ntminh@ncst.ac.vn)<br />
ABSTRACT: A new digenean is described and a second species is redescribed from the colubrid rear-fanged<br />
water snakes Enhydris chinensls (Gray) and Enhydris phunbea (Boie) captured from several regions in Vietnam<br />
during 1996-1998. Singhiatrema vietnamensis sp. n. (Ommatobrephidae) from the small intestine of both snakes<br />
is characterized by the extent of the ceca, the position of the vitellaria, the size of the eggs, and the host. Szidatia<br />
taiwanensis (Fischthal and Kuntz, 1975) comb. n. (Cyathocotylidae) is redescribed from the holotype and specimens<br />
from the gallbladder of both snakes. The species is transferred from the genus Mesostephanoides primarily<br />
because it does not have a large cirrus that is spined; it is characterized by the shape of the seminal vesicle,<br />
length of the ceca, body size relative to the forebody, number of testes in hindbody, egg size, size of tribocytic<br />
organ, and the infection site in the host. Concerning the classification of Singhiatrema Simha within Echinostomatiformes,<br />
we consider Singhiatrematinae Simha a junior synonym of Ommatobrephinae Poche. We discuss<br />
the classification of Gogatea Lutz and Szidatia Dubois and consider Gogatinae Mehra a junior synonym of<br />
Szidatiinae Dubois. The use of different fixation methods can produce artifacts characteristic at the generic level.<br />
KEY WORDS: Singhiatrema vietnamensis sp. n., Ommatobrephidae, Szidatia taiwanensis comb, n., Cyathocotylidae,<br />
fixation artifacts, Enhydris chinensis, Enhydris plumbea, Colubridae, snakes, Vietnam.<br />
Two unrelated digeneans, a new intestinal ommatobrephid<br />
and a gallbladder cyathocotylid,<br />
each infecting 2 species of rear-fanged colubrid<br />
water snakes (Enhydris chinensis (Gray, 1842)<br />
and Enhydris plumbea (Boie, 1827)) from Vietnam,<br />
are herein described. The life history of<br />
neither parasite is known. The new ommatobrephid<br />
is most similar to species of Singhiatrema<br />
Simha, 1954, which are usually found in the intestines<br />
of water snakes. Exceptions include<br />
Singhiatrema najai Chattopadhyaya, 1967, from<br />
the intestine of the Indian cobra Naja naja (Linnaeus,<br />
1758), and Singhiatrema lali Chakrabarti,<br />
1967, from the intestine of a freshwater emydid<br />
turtle (Hardella thurgii (Gray, 1831)) (Chattopadhyaya,<br />
1967). All prior known species are<br />
reported from the Indian subcontinent only.<br />
The gallbladder cyathocotylid that we found<br />
in Vietnam also occurs in one of the same hosts<br />
in Taiwan. It exhibits a relationship with Szidatia<br />
joyeuxi (Hughes, 1929), which infects the intestine<br />
of the colubrid water snake Matrix maura<br />
(Linnaeus, 1758) (as Tropidonotus viperinus<br />
(Sonnini and Latreille, 1802)) in an oasis in Tunis,<br />
Morocco, and has a cercaria that is shed<br />
from the freshwater snail Melanopsis sp. and de-<br />
Corresponding author.<br />
219<br />
velops in the leg muscles of the frog Rana rudibunda<br />
Pallas, 1771 (as Rana esculenta rudibunda;<br />
see Langeron, 1924; Hughes, 1929; Dubois,<br />
1938).<br />
Materials and Methods<br />
The second and third authors captured a total of 43<br />
specimens of E. chinensis and 51 specimens of E.<br />
plumbea in Ha Noi, Nam Ha, Thai Binh, Na Nam,<br />
Nam Dinh, Hai Duong, and Hai Phong provinces (Red<br />
River Delta) in Vietnam. They collected digeneans live<br />
from the intestines and gallbladders of both snakes<br />
from all the provinces. For the first collections, they<br />
relaxed the specimens in distilled water, then placed<br />
them in physiological saline, and finally fixed them in<br />
a cold 5% formalin solution. However, specimens collected<br />
later were used for the descriptions. These were<br />
placed directly into saline, then fixed with near boiling<br />
water without coverslip pressure, and finally pipetted<br />
into 5% formalin. Additional unmeasured specimens<br />
were fixed under pressure for examination of specific<br />
features. Whole mounts of worms were prepared by<br />
staining specimens with either carmine or Van<br />
Cleave's hematoxylin with additional Ehrlich's hematoxylin.<br />
These were dehydrated, cleared with clove oil,<br />
and mounted on slides with Canada balsam. Measurements<br />
are given for the holotype, followed in parentheses<br />
by the range of measurements of each feature<br />
derived from specimens heat-fixed without pressure.<br />
Measurements are given in micrometers unless otherwise<br />
stated. Specimens were deposited in the United<br />
<strong>State</strong>s National Parasite Collection (USNPC), Belts-<br />
Copyright © 2011, The Helminthological Society of Washington
220 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
ville, Maryland, U.S.A., and the Harold W. Manter<br />
Laboratory (HWML) of the University of Nebraska<br />
<strong>State</strong> Museum, Lincoln, Nebraska, U.S.A.<br />
Description<br />
Results<br />
Ommatobrephidae Poche, 1926<br />
Singhiatrema vietnamensis sp. n.<br />
(Figs. 1 and 2)<br />
Based on 5 specimens: Ommatobrephinae<br />
Poche, 1926. Body pyriform, elongate, 2.56 mm<br />
(2.41-2.61 mm) long, 0.76 mm (0.76-0.91 mm)<br />
wide at maximum width near posterior end of<br />
body, lacking tegumental spines but possessing<br />
minute interrupted longitudinal striae. Oral sucker<br />
with subterminal mouth, 170 (170—207) long,<br />
189 (189-217) wide. Head crown having single<br />
row of 22—23 spines 31—56 long, arranged 11—<br />
12 per side; row interrupted dorsally and ventrally.<br />
Prepharynx short, less than % length of<br />
pharynx; pharynx oval, 95 (95-106) long, 78<br />
(78-106) wide. Esophagus 640 (547-640) long,<br />
89 (80—105) wide. Ceca extending beyond posterior<br />
extent of testes. Acetabulum 290 (285-<br />
329) long, 379 (363-407) wide, lying between<br />
anterior % and anterior J/2 of body. Ratio of<br />
widths of oral sucker to acetabulum 1:1.8-2.0.<br />
Testes 2, weakly (incompletely) lobed, lying<br />
opposite, located near terminal end of body, with<br />
anterior margins diverging from each other; left<br />
testis 292 (285-340) long, 217 (200=234) wide;<br />
right testis 279 (251-335) long, 234 (195-234)<br />
wide. Cirrus sac oval, 179 (179-227) long, 102<br />
(102—110) wide, medial, lying between cecal bifurcation<br />
and acetabulum, containing large bipartite<br />
seminal vesicle, short pars prostatica, and<br />
short muscular ejaculatory duct; pars prostatica<br />
a short duct surrounded by prostatic cells at anterior<br />
of sac; external seminal vesicle lacking.<br />
Ovary pretesticular, spherical, 73 (73-95) in<br />
diameter, slightly dextral in 4 of 5 specimens,<br />
slightly sinistral in 1 specimen. Oviduct communicating<br />
with Laurer's canal, then forming<br />
ootype; Laurer's canal straight, directed dorsally<br />
from region of ootype, opening on dorsal surface<br />
at level of ovary; ootype surrounded by<br />
Mehlis' gland; Mehlis' gland compact, usually<br />
larger than ovary, consisting of relatively small<br />
cells; ootype receiving relatively long common<br />
vitelline duct from vitelline reservoir, communicating<br />
with uterus; uterus intercecal, postacetabular,<br />
with proximal portion a uterine seminal<br />
Copyright © 2011, The Helminthological Society of Washington<br />
receptacle and with distal portion a metraterm;<br />
metraterm thick walled, approximately length of<br />
cirrus sac, sinistral to cirrus sac, entering genital<br />
atrium anteriorly; genital atrium relatively small,<br />
ventral to anterior portion of cirrus sac in heatkilled<br />
specimens (directed anteriorly in coldkilled<br />
specimens when fixed under pressure with<br />
cirrus sac displaced anteriorly); genital pore<br />
opening medially or submedially near anterior<br />
region of cirrus sac in heat-killed specimens<br />
(near anterior acetabular margin in contracted<br />
specimens). Vitellarium consisting of 2 lateral<br />
bands of irregularly shaped follicles; bands lying<br />
ventral to ceca between posterior level of acetabulum<br />
and middle of testes, communicating to<br />
vitelline reservoir by left and right transverse<br />
main collecting channel; left channel about 195<br />
long, right channel about 223-280 long; vitelline<br />
reservoir approximately 45—56 long, 111-195<br />
wide. Eggs 95-117 long, 61-75 wide, with<br />
thickened knob at posterior end, with eyespots<br />
visible in miracidia of some specimens; eyespots<br />
2, lightly pigmented in developing miracidia,<br />
darkly pigmented and fused in developed miracidia.<br />
Excretory vesicle Y-shaped, with triangular<br />
posterior bladder; main stem slender, medial,<br />
concealed by overlapping lobes of testes, extending<br />
anteriorly from bladder and bifurcating<br />
at anterior margins of testes; arms reaching anteriorly<br />
to approximate level just posterior to<br />
pharynx; excretory pore subterminal, opening<br />
on dorsal surface.<br />
Taxonomic summary<br />
TYPE HOST: Enhydris chinensis (Gray,<br />
1842), rear-fanged water snake or Chinese water<br />
snake (Colubridae). Other host: Enhydris plumbea<br />
(Boie, 1827), rear-fanged water snake, rice<br />
paddy snake, or plumbeous water snake (Colubridae).<br />
TYPE LOCALITY: Ha Noi Province, Vietnam.<br />
Other localities: throughout Red River Delta, Vietnam,<br />
in Nam Ha, Thai Binh, Nam Ha, Nam<br />
Dinh, Hai Duong, and Hai Phong provinces.<br />
INFECTION SITE: Small intestine.<br />
PREVALENCE AND INTENSITY OF INFECTION: Ten<br />
of 43 specimens of E. chinensis (23%) each<br />
hosted 1-5 individual worms; 9 of 51 specimens<br />
of E. plumbea (18%) each hosted 1—3 worms.<br />
SPECIMENS DEPOSITED: Holotype USNPC No.<br />
90037; paratypes USNPC No. 90038, HWML<br />
Nos. 15388 (E. chinensis), 15389 (E. plumbea).
CURRAN ET AL.—TWO VIETNAMESE SNAKE DIGENEANS 221<br />
Figures 1-6. 1. Ventral view of holotype of Singhiatrema vietnamensis sp. n.; scale bar = 300 u.m. 2.<br />
Ventral view of specimen of 5. vietnamensis exposed to fresh water and slight pressure prior to and during<br />
cold fixation; scale bar = 300 (mm. 3. Ventral view of Szidatia taiwanensis; scale bar = 200 \nm. 4. Ventral<br />
view of S. taiwanensis showing detail of tribocytic organ; scale bar = 200 (Jim. 5. Lateral view of 5.<br />
taiwanensis; scale bar = 200 (Jim. 6. Ventral view of 5. taiwanensis exposed to fresh water prior to cold<br />
fixation; scale bar = 275 (xm.<br />
Copyright © 2011, The Helminthological Society of Washington
222 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
ETYMOLOGY: This species is named for its<br />
type locality, Vietnam.<br />
Remarks<br />
Singhiatrema vietnamensis is consistent with<br />
members of Ommatobrephidae because it has a<br />
preacetabular cirrus sac with an internal seminal<br />
vesicle; large embryonated eggs, with some containing<br />
an oculate miracidium; and opposite incompletely<br />
lobed testes located in the posterior<br />
region of the body, often with axes diverging<br />
anteriorly. The species belongs in Ommatobrephinae<br />
rather than Parorchiinae Lai, 1936, because<br />
the collar of spines is interrupted dorsally<br />
and ventrally as opposed to being arranged in a<br />
continuous row. In addition, the tegument is<br />
smooth rather than spinous, an external seminal<br />
vesicle is absent, spines do not occur on the intromittent<br />
organ, and the specimens are parasites<br />
in reptiles rather than birds. The 2 genera within<br />
Ommatobrephinae, Singhiatrema and Ommatobrephus<br />
Nicoll, 1914, are distinguished by the<br />
presence of a dorsally interrupted row of collar<br />
spines in the former and the absence of the collar<br />
spines in the latter. The presence of a dorsally<br />
and ventrally interrupted row of 22-23 collar<br />
spines in 5. vietnamensis enables us to assign<br />
the worms to the genus Singhiatrema. Singhiatrema<br />
was originally unveiled to science in an<br />
oral presentation and abstract at the Forty-first<br />
Session of the Indian Science Congress (Simha,<br />
1954). The genus was described in more detail<br />
by Simha (1958) and placed in Echinostomatidae<br />
Poche, 1926, by the author without a subfamily<br />
designation. Singhiatrema singhia Simha,<br />
1954, from a colubrid ratsnake (Ptyas mucosus<br />
(Linnaeus, 1758)) from Hyderabad in southern<br />
India, was designated the type species (Simha,<br />
1954). Two other species, Singhiatrema longifurca<br />
Simha, 1958, and Singhiatrema hyderabadensis<br />
Simha, 1958, parasitize another colubrid<br />
water snake, the checkered keelback Xenochrophis<br />
piscator (Schneider, 1799) (as Tropidonotus<br />
piscator}, from the same locality<br />
(Simha, 1958). Three other Indian species have<br />
since been added to the genus. Singhiatrema najai<br />
parasitizes the Indian cobra (N. najd) in Hyderabad,<br />
S. lali parasitizes the turtle H. thurgii<br />
in Lucknow, and Singhiatrema piscatora Dwivedi,<br />
19<strong>68</strong>, parasitizes the water snake X. piscator<br />
in Chhindwara (Chakrabarti, 1967; Chattopadhyaya,<br />
1967; Dwivedi, 19<strong>68</strong>).<br />
Singhiatrema vietnamensis differs from its<br />
Copyright © 2011, The Helminthological Society of Washington<br />
congeners, with the exception of S. lali, by having<br />
ceca that extend to the posterior region of<br />
the body and vitelline follicles that lie ventral<br />
and lateral to the ceca in bands stretching from<br />
the posterior margin of the acetabulum to the<br />
midlevel of the testes. Singhiatrema vietnamensis<br />
differs from S. lali by having larger eggs<br />
(103-119 |xm long by 45-56 jxm wide vs. 65-<br />
87 |xm long by 34-39 |xm wide), ceca that reach<br />
beyond the posterior level of the testes rather<br />
than to their midlevel, longer collar spines (31-<br />
56 (Jim rather than 7-9 jxm), and a snake rather<br />
than a turtle definitive host. Yamaguti (1971) reported<br />
24 collar spines for S. lali; however,<br />
Chakrabarti (1967) did not report the number of<br />
collar spines in the description of S. lali, and the<br />
illustration of the species did not permit the collar<br />
spines to be counted. Specimens of S. lali<br />
were apparently never deposited in a lending<br />
museum. We report 22-23 collar spines in our<br />
specimens of 5. vietnamensis. Three of 5 of our<br />
specimens had 22 collar spines and the remaining<br />
2 had 23 collar spines. It is possible that<br />
some specimens could lose spines in life or handling,<br />
but biological variation probably exists.<br />
Initially, we obtained 4 specimens of S. vietnamensis<br />
that had been placed in fresh water<br />
prior to their fixation with slight pressure in unheated<br />
formalin (see Fig. 2). The overall shape<br />
of these specimens, as well as their measurements,<br />
varied dramatically from those specimens<br />
fixed with heat and used in the above description.<br />
Specimens subjected to fresh water and<br />
pressure had an oval rather than pyriform body<br />
shape, and their width was greater (1.0-1.6 mm<br />
vs. 0.75-0.91 mm). The pharynx was swollen<br />
(131-158 |xm long by 127-140 jxm wide vs. 95-<br />
106 (Jim long by 78-106 |xm wide), and the<br />
esophagus was contracted in length but swollen<br />
in width (285-415 (xm long by 77-203 |xm wide<br />
vs. 547-640 (xm long by 80-105 |xm wide). In<br />
addition, the ceca were contracted slightly,<br />
reaching to the posterior level of the testes rather<br />
than beyond the posterior level of the testes, and<br />
the eggs were swollen or collapsed and therefore<br />
larger in whole mounts (109—117 |xm long by<br />
81-95 urn wide vs. 103-119 |xm long by 45-56<br />
(xm wide). The long side of the oval cirrus sac<br />
was oriented laterally rather than vertically, with<br />
the pore on the sinistral end rather than at the<br />
anterior end, and the testes were larger (411-560<br />
|xm long by 258-339 |xm wide compared with<br />
250-340 |xm long by 195-234 |xm wide). If the
methods of fixation had not been known, these<br />
differences would be great enough to suspect<br />
different species.<br />
Cyathocotylidae Poche, 1926<br />
Szidatia taiwanensis<br />
(Fischthal and Kuntz, 1975) comb. n.<br />
(Figs. 3-6)<br />
Redescription<br />
Based on holotype and 9 specimens from Vietnam:<br />
Szidatiinae Dubois, 1938. Body bipartite;<br />
forebody pyriform, 0.85 mm (0.81-1.15 mm)<br />
long, not accounting for slight curl; hindbody<br />
conical; entire body 1.20 mm (1.03-1.57 mm)<br />
long, 0.57 mm (0.47-0.93 mm) wide at maximum<br />
width just posterior to midforebody. Ratio<br />
of forebody to total body length 1:1.39 (1:1.27-<br />
1.36). Tegument with spines over entire forebody,<br />
with few or no spines on hindbody, with<br />
numerous minute papillae near posterior end.<br />
Oral sucker with subterminal mouth, (90-119)<br />
long, 119 (99-144) wide. Prepharynx very<br />
short. Pharynx oval, (60-75) long, (60-80)<br />
wide. Esophagus 94 (67-149) long, 31 (23-40)<br />
wide. Ceca bifurcating at about anterior Vs of<br />
forebody, extending to level of posterior margin<br />
of ovary. Acetabulum 82 (62-95) long, 98 (85-<br />
1 12) wide, situated between anterior % and anterior<br />
l/2 of forebody, always smaller than oral<br />
sucker. Tribocytic organ oval, protruding slightly<br />
from ventral surface, 327 (283-447) long, -250<br />
(246-423) wide, with medial-longitudinal slit<br />
with basal layer of dark-staining cells.<br />
Testes 2, oblique; anterior testis spanning both<br />
forebody and hindbody, -80 (149-213) long,<br />
124 (114-169) wide; posterior testis in hindbody<br />
91 (144-199) long, 142 (129-229) wide. Cirrus<br />
sac in posterior end of body, club-shaped, 410<br />
(332-362) long, 65 (60-73) wide, extending anteriorly<br />
to middle of anterior testis, containing<br />
internal seminal vesicle, pars prostatica with<br />
small prostatic cells, and short unspined ejaculatory<br />
duct (not true cirrus); internal seminal<br />
vesicle 241 (206-339) long, 45 (57-110) wide<br />
at thickest portion; pars prostatica 1 1 1 (43-198)<br />
long, —30 (23-25) wide, sinuous in some specimens,<br />
surrounded externally by free gland cells;<br />
ejaculatory duct 45 (24-74) long, 20 (8-14)<br />
wide, thin walled, eversible, surrounded by<br />
gland cells.<br />
Ovary nearly spherical, 85 (65-129) long, 75<br />
(70—99) wide. Vitellarium comprised of 2 pairs<br />
CURRAN ET AL.—TWO VIETNAMESE SNAKE DIGENEANS 223<br />
of lateral fields of follicles underlying tribocytic<br />
organ, with the 2 on each side overlying each<br />
other, ventral to ovary and testes; fields not confluent<br />
anteriorly; right field totaling —12-14<br />
(16-20) follicles; left field totaling -14 (16-23)<br />
follicles; follicles spherical to ovate, ranging 37—<br />
43 (50-99) long, >50 (55-124) wide. Vitelline<br />
reservoir lying posterior to ovary and between<br />
testes, 145 (119-149) long, 80 (78-144) wide,<br />
(159-167) thick (thickness measured from 2 laterally<br />
mounted specimens). Uterus reaching anteriorly<br />
to near level of anterior extent of vitellarium;<br />
distal portion an indistinct metraterm;<br />
metraterm slightly longer than cirrus sac (coiled<br />
in holotype and some other specimens), demarcated<br />
by transverse muscular band, surrounded<br />
by lining of free gland cells, emptying into genital<br />
atrium separate from ejaculatory duct; genital<br />
atrium an open funnel in distal portion of<br />
body, surrounded by gland cells. Eggs 5 (4-12)<br />
in number, 150 (140-169) long, 77 (80-99)<br />
wide, with indistinct operculum.<br />
Excretory vesicle V-shaped; arms united both<br />
anteriorly and medially; anterior junction at level<br />
of and dorsal to midesophagus; medial junction<br />
at level of anterior extent of tribocytic organ,<br />
ventral to ceca, leading to small bladder<br />
associated with base of acetabulum; excretory<br />
pore subterminal, opening ventral to and separate<br />
from constricted genital atrium.<br />
Taxonomic summary<br />
HOSTS: Enhydris chinensis (Gray, 1842),<br />
rear-fanged water snake or Chinese water snake<br />
(Colubridae), and Enhydris plumbea (Boie,<br />
1827), rear-fanged water snake, rice paddy<br />
snake, or plumbeous water snake (Colubridae).<br />
LOCALITIES: Ha Noi, Nam Ha, Thai Binh,<br />
Ha Nam, Nam Dinh, Hai Duong, and Hai Phong<br />
provinces in Vietnam.<br />
INFECTION SITE: Gallbladder (holotype USNPC<br />
No. 73148 from Taiwan in E. chinensis from<br />
intestine).<br />
PREVALENCE AND INTENSITY OF INFECTION: Sixteen<br />
of 43 specimens from intestine of E. chinensis<br />
(37%) each hosted 1-9 individual worms;<br />
24 of 51 specimens of E. plumbea (47%) each<br />
hosted 1-9 worms.<br />
SPECIMENS DEPOSITED: Voucher specimens<br />
USNPC Nos. 90039 and 90040 (E. chinensis),<br />
HWML No. 15390 (2 slides, E. chinensis).<br />
Copyright © 2011, The Helminthological Society of Washington
224 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Remarks<br />
Fischthal and Kuntz (1975) described Mesostephanoid.es<br />
taiwanensis from the small intestine<br />
of E. chinensis from Taipei Prefecture in<br />
Taiwan on the basis of a single specimen. We<br />
examined that specimen, USNPC No. 73148,<br />
and, even though it was not from the gallbladder,<br />
we considered our specimens from Vietnam<br />
conspecific with it. Data for the holotype fit<br />
those for our specimens. We, however, interpret<br />
differently the terminal genitalia described and<br />
illustrated by Fischthal and Kuntz (1975) in their<br />
Figure 15. Perhaps they misinterpreted the nonillustrated<br />
muscular looping of the metraterm as<br />
spines. In any event, the cirrus sac of the holotype<br />
contained a club-shaped seminal vesicle<br />
measuring about 260 u,m long, a straight pars<br />
prostatica 113 u,m long, and a short ejaculatory<br />
duct 45 fjim long. The metraterm coiled once<br />
prior to descending to the genital atrium. We observed<br />
spines in none of these features. Because<br />
of the lack of a spined cirrus and lack of anterior<br />
confluence of the bands of vitelline follicles, we<br />
consider that the species belongs to Szidatia Dubois,<br />
1938, as Szidatia taiwanensis (Fischthal and<br />
Kuntz, 1975) comb. n.<br />
Dubois (1951) differentiated the monotypic<br />
genus Mesostephanoides Dubois, 1951, from<br />
other cyathocotylid genera on the basis of Mesostephanoides<br />
burmanicus (Chatterji, 1940),<br />
having a forebody to total body length ratio that<br />
we estimate as 1:1.14-1.18, a spined cirrus 300<br />
(Jim long, and vitelline follicles confluent anteriorly.<br />
We consider the first 2 features of questionable<br />
generic significance, and an examination<br />
of M. burmanicus and related species may<br />
show that Mesostephanoides is a synonym of<br />
Gogatea Lutz, 1935, a genus with species having<br />
anteriorly confluent vitelline follicles.<br />
Szidatia taiwanensis resembles its 2 congeners<br />
in that the vitelline follicles are in separate<br />
lateral fields, as opposed to being in a single<br />
confluent field underlying the tribocytic organ as<br />
reported for species of Gogatea, the only other<br />
genus in Szidatiinae. Szidatia taiwanensis most<br />
closely resembles Szidatia nemethi Dollfus,<br />
1953, from the water snake TV. maura (as N. viperina)<br />
from Charrat, Morocco, but differs from<br />
this species by having a much smaller tribocytic<br />
organ (283—447 jjim long by 246-423 |xm wide<br />
vs. 885 (Jim long by 765 u,m wide), larger eggs<br />
(140-169 jxm long by 80-99 (Jim wide vs. 105<br />
Copyright © 2011, The Helminthological Society of Washington<br />
(xm long by 55 (Jim wide), and a club-shaped<br />
rather than a coiled seminal vesicle. Szidatia taiwanensis<br />
differs from S. joyeuxi, the type and<br />
remaining species in the genus, by having a<br />
longer esophagus (67-149 |JLm compared with<br />
40 |xm), ceca that reach only to the level of the<br />
anterior testis rather than beyond the posterior<br />
testis, an acetabulum that lies between the middle<br />
and anterior % of the forebody rather than<br />
exactly in the middle, and oblique rather than<br />
tandem testes; the anterior testis is almost entirely<br />
within the forebody rather than in the<br />
hindbody, the seminal vesicle is club-shaped<br />
rather than coiled, and the eggs are larger (140-<br />
169 (xm long by 80-99 (Jim wide vs. 100 (Jim<br />
long by 70 (xm wide). The minute papillae on<br />
the tegument at the posterior end, too small to<br />
illustrate to scale, probably serve a sensory<br />
function during mating or egg deposition. The<br />
intestine, the infection site of the specimen reported<br />
by Fischthal and Kuntz (1975), may have<br />
resulted from postmortem migration from the<br />
gallbladder. A few specimens of other species<br />
normally found in the gallbladder occasionally<br />
occur in the intestine normally. We do not think<br />
the difference in sites based on 1 specimen is<br />
significant.<br />
As with the echinostomate specimens of S.<br />
vietnamensis, the initial specimens of S. taiwanensis<br />
had been placed in fresh water prior to<br />
fixation in unheated formalin (see Fig. 6). Unlike<br />
those of S. vietnamensis, these specimens<br />
were not fixed under any pressure; nevertheless,<br />
their measurements and features varied dramatically<br />
from those obtained from specimens fixed<br />
with heat and used for the above description.<br />
Hindbody length was generally shorter (146-<br />
506 (Jim vs. 326—762 (Jim), and both the anterior<br />
and posterior testes occurred entirely in the forebody<br />
of specimens exposed to water, whereas<br />
the posterior testis was always in the hindbody<br />
of heat-killed specimens. The cirrus sac was<br />
generally straighter in osmotically stressed specimens<br />
and more bent or curled in the region of<br />
the pars prostatica in heat-killed specimens. In<br />
addition, the tribocytic organ was greatly enlarged<br />
(492-650 (Jim long by 520-<strong>68</strong>5 |xm wide<br />
vs. 283-447 fxm long by 245-423 ujn wide) in<br />
the stressed specimens. The most important difference<br />
due to fixation techniques pertained to<br />
the configuration of the vitellarium. In the<br />
stressed specimens, vitelline follicles appeared<br />
confluent anteriorly (horseshoe-shaped distribu-
tion) but were in independent lateral bands in<br />
heat-killed specimens. Because the presence or<br />
absence of an anterior vitelline confluence differentiates<br />
Gotatea from Szidatia, and the appearance<br />
of confluence can be influenced in<br />
cold-killed or freshwater-soaked specimens, the<br />
methods of fixation clearly have a bearing on<br />
taxonomic interpretations.<br />
Discussion<br />
Classification of Singhiatrema<br />
The position of Singhiatrema within Echinostomatiformes<br />
La Rue, 1957, continues to be debated<br />
by taxonomists. Ommatobrephinae Poche,<br />
1926, was created for Ommatobrephus and<br />
Singhiatrema, and Parorchiinae Lai, 1936, was<br />
included in Ommatobrephidae on the basis of<br />
the generic level character of the collar spines<br />
(see Simha and Chattopadhyaya, 1967). Singhiatrematinae<br />
Simha, 1962, was later created to<br />
house Singhiatrema, presumably because the author<br />
considered the presence of collar spines to<br />
be of taxonomic importance at the subfamiliar<br />
rather than generic level. These changes resulted<br />
in Ommatobrephidae containing Ommatobrephinae,<br />
Parorchiinae, and Singhiatrematinae, each<br />
containing a single genus. Members of Ommatobrephinae<br />
and Singhiatrematinae differ only in<br />
the presence or absence of a single row of collar<br />
spines. We consider the presence or absence of<br />
collar spines to represent a generic feature only<br />
and, therefore, consider Singhiatrematinae a junior<br />
subjective synonym of Ommatobrephinae.<br />
Yamaguti (1971) considered Parorchiinae a subfamily<br />
in Philophthalmidae Travassos, 1918. We<br />
agree with Yamaguti in returning Parorchiinae<br />
to Philophthalmidae because members of that<br />
subfamily do not possess characters consistent<br />
with Ommatobrephidae. Most notably, parorchiines<br />
have tegumental spines, a conspicuous prepharynx,<br />
an external seminal vesicle, a spined<br />
cirrus, opposite testes that do not diverge anteriorly,<br />
and life cycles that involve birds and estuarine<br />
molluscs. Although no knowledge of the<br />
larval stages of species in Singhiatrema or Ommatobrephus<br />
is available, all known adults of<br />
species in these genera infect freshwater or terrestrial<br />
reptilian hosts, indicating a freshwater<br />
rather than an estuarine life cycle.<br />
Classification of Szidatia<br />
Contrary opinions regarding the status of key<br />
generic features of Gogatea and Szidatia have<br />
CURRAN ET AL.—TWO VIETNAMESE SNAKE DIGENEANS 225<br />
formed the basis for debates over whether the 2<br />
cyathocotylid subfamilies Szidatiinae and Gogatinae<br />
Mehra, 1947, should be synonymized<br />
(see Dubois, 1938, 1951, 1953; Mehra, 1947;<br />
Sudarikov, 1962; Yamaguti, 1971). Both subfamilies<br />
contain only a single genus, but the<br />
above discussion of Mesostephanoides should<br />
be noted.<br />
Lutz (1935) erected Gogatea and created the<br />
combination Gogatea serpentum (Gogate, 1932)<br />
for Prohemistomum serpentum Gogate, 1932, a<br />
parasite in the intestine of the colubrid snake X.<br />
piscator (as Natrix piscator) from the Union of<br />
Myanmar (as Burma). Lutz (1935) included Gogatea<br />
in his new subfamily Prohemistominae<br />
Lutz, 1935. Szidat (1936) added the new combination<br />
Gogatea joyeuxi (Hughes, 1929) for the<br />
cyathocotylid previously considered Prohemistomum<br />
joyeuxi (Hughes, 1929) from the colubrid<br />
water snake Natrix natrix scutata Pallas,<br />
1771 (as Tropidontus natrix persa), by Joyeux<br />
and Baer (1934). That species developed in the<br />
snake when fed diplostomula consistent with diplostomula<br />
described by Hughes (1929) from<br />
Tunis, Morocco (Joyeux and Baer, 1934). Gogatea<br />
indicum Mehra, 1947, was subsequently<br />
described from X. piscator in India. Dubois<br />
(1938) observed that the vitellarium of G. joyeuxi<br />
consisted of 2 distinct rows of vitelline follicles,<br />
1 on either side of the tribocytic organ.<br />
The vitellarium of other species in Gogatea consists<br />
of a confluent arch of follicles, lying dorsal<br />
to the tribocytic organ. Dubois (1938) erected<br />
Szidatia Dubois, 1938, on the basis of the differing<br />
configuration of the vitellaria and created<br />
the new combination S. joyeuxi for G. joyeuxi.<br />
Dubois (1938) also created the subfamily Szidatinae,<br />
emended as Szidatiinae by Yamaguti<br />
(1958), to house both Szidatia and Gogatea.<br />
Mehra (1947) did not consider that the difference<br />
in vitelline configuration warranted generic<br />
distinction and rejected Szidatia and therefore<br />
Szidatiinae. Instead, he created Gogatinae to<br />
contain all species of Gogatea, with G. serpentum<br />
as the type species. Dollfus (1953), Sudarikov<br />
(1962), and Yamaguti (1971) all accepted<br />
the generic distinctions established by Dubois<br />
(1938) and considered Gogatinae a junior synonym<br />
for Szidatiinae, retaining G. serpentum<br />
and S. joyeuxi as the type species for their respective<br />
genera. We concur with these authors<br />
and think that the different configuration of the<br />
Copyright © 2011, The Helminthological Society of Washington
226 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
vitellarium can be used as an informative generic<br />
level taxonomic distinction.<br />
Fixation techniques<br />
A wide variety of techniques has been used<br />
by parasitologists to fix specimens. Most of<br />
those techniques yield satisfactory and consistent<br />
results; however, those that produce results<br />
inconsistent with other procedures should be<br />
avoided. We advocate collecting digeneans by<br />
initially placing them live in physiological saline<br />
solution (7.7—8.5 g NaCl per liter of distilled water).<br />
They should never be exposed to distilled<br />
or tap water while alive. Unless specimens are<br />
to be used for ultrastructural or other specific<br />
puiposes, they should be killed rapidly with<br />
heat. They can be killed by a flame under a slide<br />
with worms in a small amount of saline or by<br />
pouring a relatively large volume of hot or boiling<br />
saline or tap water over specimens immersed<br />
in little or no saline. The specimens then should<br />
be transferred, without touching them with forceps,<br />
into 5—10% buffered formalin solution<br />
soon after being killed. Killing with hot formalin<br />
solution or other fixatives is acceptable but produces<br />
harsh fumes. Killing with heat produces<br />
consistently fixed specimens, ideal for making<br />
comparative measurements. Fixation with pressure<br />
may be useful for examining certain organ<br />
systems such as the female reproductive complex<br />
or the terminal genitalia, but specimens<br />
fixed under pressure should be used with care<br />
for taxonomic purposes because the entire specimen<br />
or specific structures may be distorted.<br />
Comparison of heat-killed specimens with<br />
specimens bathed in fresh water and then coldkilled<br />
revealed differences. Specimens of S. vietnamensis<br />
that were osmotically stressed prior to<br />
fixation were a different shape, they were wider,<br />
the pharynx and esophagus were distorted, the<br />
ceca were slightly shorter, the testes were larger,<br />
and the cirrus sac was oriented differently. Any<br />
one of these conditions might be used to misidentify<br />
a species, incorrectly describe specimens,<br />
or provide misleading information.<br />
Stressed specimens of 5. taiwanensis exhibited<br />
some distortions at the specific level, but more<br />
importantly, the confluent vitelline follicles represent<br />
a generic distinction for Gogatea. Lack of<br />
knowledge about the methods used on specimens<br />
for descriptions of some species of Gogatea<br />
and Szidatia shows the need to reevaluate<br />
those species with heat-killed specimens. Until<br />
Copyright © 2011, The Helminthological Society of Washington<br />
such material is available, we prefer to treat the<br />
genera separately.<br />
Acknowledgments<br />
We thank Cathy Schloss and Kristine Wilkie<br />
of the Gulf Coast Research Laboratory for helping<br />
with some of the literature. We also thank<br />
Eric Hoberg and Pat Pilitt from the United <strong>State</strong>s<br />
National Parasite Collection, Beltsville, Maryland,<br />
for the loan of a specimen and Dr. Richard<br />
Heckmann of Brigham Young University for his<br />
interest in the project. Partial support for this<br />
study was provided by International Paper.<br />
Literature Cited<br />
Chakrabarti, K. K. 1967. On a new echinostome<br />
(Singhiatrema lali n. sp.) from the intestine of a<br />
turtle. Science and Culture 33:407-408.<br />
Chattopadhyaya, D. R. 1967. On a new trematode of<br />
the genus Singhiatrema Simha, 1954, from the<br />
cloaca of the Indian cobra in Hyderabad. Indian<br />
Journal of Helminthology 18:45-49.<br />
Dollfus, R. P. 1953. Miscellanea helminthologica<br />
Maroccana VII. Les Szidatia de Matrix viperina<br />
(Latreille, 1802) [Trematoda Digenea]. Archives<br />
de 1'Institut Pasteur du Maroc 4:505-512.<br />
Dubois, G. 1938. Monographic des Strigeida (Trematoda).<br />
Memoires de la Societe Neuchateloise des<br />
Sciences Naturelles 6:1—535.<br />
. 1951. Nouvelle cle de determination des groupes<br />
systematiques et des genres de Strigeida<br />
Poche (Trematoda). Revue Suisse de Zoologie<br />
58(39):639-691.<br />
-. 1953. Liste Systcmatique des Strigeida (Trematoda)<br />
de 1'Inde. Pages 77-88 in J. Dayal and K.<br />
Singh, eds. Thapar, G. S., Commemoration Volume.<br />
University of Lucknow, Lucknow, India.<br />
Dwivedi, M. P. 19<strong>68</strong>. On a new species of the genus<br />
Singhiatrema Simha, 1954 (Trematoda: Echinostomidae).<br />
Indian Journal of Helminthology 19:<br />
141-144.<br />
Fischthal, J. H., and R. E. Kuntz. 1975. Some trernatodes<br />
of amphibians and reptiles from Taiwan.<br />
Proceedings of the Helminthological Society of<br />
Washington 42:1-13.<br />
Hughes, R. C. 1929. Studies on the trematode family<br />
Strigeidae (Holostomidae) No. XIV. Two new species<br />
of diplostomula. Occasional Papers of the<br />
Museum of Zoology, University of Michigan 202:<br />
1-29 + 1 pi.<br />
Joyeux, C., and J. G. Baer. 1934. Sur un trematode<br />
de couleuvre. Revue Suisse de Zoologie 41:203-<br />
215.<br />
Langeron, M. 1924. Recherches sur les cercaires des<br />
piscines de Gafsa et enquete sur la bilharziose<br />
Tunisienne. Archives de FInstitut Pasteur de Tunis<br />
13:19-67.<br />
Lutz, A. 1935. Observances e consideracoes sobre<br />
Cyathocotylineas e Prohemistomineas. Memorias<br />
do Institute Oswaldo Cruz 30:157-182.<br />
Mehra, H. R. 1947. Studies on the family Cyathoco-
tylidae Poche: Part 2. A contribution to our<br />
knowledge of the subfamily Prohemistominae<br />
Lutz, 1935, with a discussion on the classification<br />
of the family. Proceedings of the National Academy<br />
of Sciences, India 17:1-52.<br />
Simha, S. S. 1954. On a new trematode genus Singhiatrema<br />
singhia n. g., n. s. from rat snake Ptyas<br />
(Zamenis) miicosus from Hyderabad-Deccan. Proceedings<br />
of the Indian Science Congress (1954)<br />
Part IV 41:32.<br />
. 1958. Studies on the trematode parasite of<br />
reptiles found in Hyderabad state. Zeitschrift fur<br />
Parasitenkunde 18:161-218.<br />
, and D. R. Chattopadhyaya. 1967. A discussion<br />
on the systematic position of the subfamily<br />
Singhiatreminae Simha, 1962. Indian Journal of<br />
Helminthology 18(seminar supplement):54-58.<br />
CURRAN ET AL.—TWO VIETNAMESE SNAKE DIGENEANS 227<br />
Symposium Announcement<br />
Sudarikov, V. E. 1962. Order Strigeidida (La Rue,<br />
1926) Sudarikov, 1959. Trematodes of Animals<br />
and Man. Principles of Trematodology, Academy<br />
of Sciences of the U.S.S.R. Helminthological Laboratory,<br />
Moscow 19:267-469. (In Russian.)<br />
Szidat, L. 1936. Parasiten aus Seeschwalben 1. Uber<br />
neue Cyathocotyliden aus dem Darm von Sterna<br />
hirundo L. und Sterna paradisea. Zeitschrift fur<br />
Parasitenkunde 8:285-317.<br />
Yamaguti, S. 1958. Systema Helminthum. Volume I.<br />
The Digenetic Trematodes of Vertebrates, Part I.<br />
Interscience Publishers, New York, New York.<br />
979 pp.<br />
. 1971. Synopsis of Digenetic Trematodes of<br />
Vertebrates. 2 volumes. Keigaku Publishing Company,<br />
Tokyo, Japan. 1074 pp. + 349 pis.<br />
PARASITOLOGY IN SCIENCE AND SOCIETY<br />
Sponsored by The Helminthological Society of Washington<br />
Saturday, October 27, <strong>2001</strong>, 1:00-5:00 pm<br />
S. Dillon Ripley Center of the South Quadrangle, Room 3111<br />
National Museum of Natural History of the Smithsonian Institution<br />
Washington, D.C.<br />
The 676th meeting of the Helminthological Society of Washington, to be held on October 27, <strong>2001</strong> at<br />
the Smithsonian Institution, will be a special event. The Helminthological Society of Washington, in<br />
cooperation with The American Society of Parasitologists, will conduct a symposium entitled <strong>Parasitology</strong><br />
in Science and Society. The purpose of the symposium is to assess the state of parasitology as a discipline,<br />
clearly define the role of parasitology in science, and explore ways that various parasitological and related<br />
societies and governmental agencies can work together to strengthen our discipline. Dr. Richard O'Grady,<br />
Executive Director of the American Institute of Biological Sciences, will speak on the power and importance<br />
of collaborative efforts in science. Dr. Eric P. Hoberg of the United <strong>State</strong>s Department of Agriculture's<br />
Beltsville Agricultural Research Center, will discuss the seminal importance of phylogenetic studies<br />
and taxonomic inventories and collections to contemporary parasitology. Additionally, leaders of various<br />
parasitological and related societies and governmental agencies are being asked to clearly articulate the<br />
missions and visions of their organizations as they relate to parasitology. Because each society and agency<br />
is a unique entity, we all have special strengths to offer to our broader discipline. Further, participants are<br />
being asked to identify contemporary societal issues which the discipline of parasitology can address, and<br />
indicate specific ways that their organization is, and can aid in, addressing these issues. We invite you to<br />
make every effort to attend this important meeting of the Helminthological Society of Washington.<br />
INVITED SOCIETIES, AGENCIES, AND ORGANIZATIONS<br />
American Association of Veterinary Parasitologists<br />
American Heartworm Society<br />
American Institute of Biological Sciences<br />
American Society of Parasitologists<br />
American Society of Tropical Medicine and Hygiene<br />
Canadian Zoological Society<br />
Centers for Disease Control and Prevention<br />
Entomological Society of America<br />
Helminthological Society of Washington<br />
National Institutes of Health<br />
National Science Foundation<br />
Society of Nematologists<br />
Society of Protozoologists<br />
United <strong>State</strong>s Department of Agriculture<br />
United <strong>State</strong>s Department of the Interior<br />
Wildlife Disease Association<br />
Copyright © 2011, The Helminthological Society of Washington
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 228-235<br />
Rhabdias ambystomae sp. n. (Nematoda: Rhabdiasidae) from the<br />
North American Spotted Salamander Ambystoma inaculatum<br />
(Amphibia: Ambystomatidae)<br />
YURIY KUZMIN,' VASYL V. TKACH,1-2 AND SCOTT D. SNYDER3'4<br />
1 Department of <strong>Parasitology</strong>, Institute of Zoology, Ukrainian National Academy of Sciences, 15 Bogdan<br />
Khmelnitsky Street, Kiev-30, MSP, 01601, Ukraine,<br />
2 Institute of <strong>Parasitology</strong>, Polish Academy of Sciences, Twarda Street 51/55, 00-818, Warsaw, Poland, and<br />
3 Department of Biology and Microbiology, University of Wisconsin Oshkosh, Oshkosh, Wisconsin 54901,<br />
U.S.A. (e-mail: snyder@uwosh.edu)<br />
ABSTRACT: Rhabdias ambystomae sp. n. is described on the basis of specimens found in the lungs and body<br />
cavity of the spotted salamander (Ambystoma maculatum) from northwestern Wisconsin, U.S.A. The new species<br />
differs from Rhabdias bermani in tail shape, arrangement of circumoral lips, and position of vulva, from Rhabdias<br />
tokyoensis in the morphology and size of the buccal capsule and the shape of the esophagus, and from<br />
Rhabdias americanus in the absence of pseudolabia at the cephalic extremity and the shape of the tail. Rhabdias<br />
ambystomae sp. n. is the first species of the genus described from salamanders in North America.<br />
KEY WORDS: Nematoda, Rhabdiasidae, Rhabdias ambystomae sp. n., salamanders, Ambystoma maculatum,<br />
Wisconsin, U.S.A.<br />
Nematodes of the genus Rhabdias Stiles and<br />
Hassall, 1905, are globally distributed lung parasites<br />
of amphibians and reptiles. Among amphibian<br />
hosts, the vast majority of Rhabdias species<br />
have been reported from anurans (frogs and toads),<br />
whereas only 2 species of the genus have been<br />
described from caudatans (salamanders): Rhabdias<br />
bermani Rausch, Rausch, and Atrashkevich, 1984,<br />
from the Siberian newt Salamandrella keyserlingii<br />
Dybowski, 1870, in the eastern Palearctic (Rausch<br />
et al., 1984) and Rhabdias tokyoensis Wilkie,<br />
1930, from Cynops spp. in Japan (Wilkie, 1930).<br />
In North America, Rhabdias spp. previously have<br />
been found in the lungs and body cavities of several<br />
species of salamanders (Lehmann, 1954; Dyer<br />
and Peck, 1975; Price and St. John, 1980; Coggins<br />
and Sajdak, 1982; Muzzall and Schinderle, 1992;<br />
Bolek and Coggins, 1998; Goldberg et al., 1998).<br />
These nematodes were identified as either Rhabdias<br />
sp., Rhabdias ranae Walton, 1929, or Rhabdias<br />
joaquinensis Ingles, 1935, the latter 2 species<br />
normally restricted to anuran amphibians.<br />
In the course of investigations of the helminth<br />
fauna of Wisconsin amphibians, infections by a<br />
species of Rhabdias were detected in the lungs and<br />
body cavities of 2 specimens of the spotted salamander<br />
Ambystoma maculatum (Shaw, 1802).<br />
Morphological examination revealed these worms<br />
Corresponding author.<br />
228<br />
Copyright © 2011, The Helminthological Society of Washington<br />
to represent a new species of the genus Rhabdias.<br />
This species is described herein as Rhabdias ambystomae<br />
sp. n.<br />
Materials and Methods<br />
Amphibians were collected from a roadside wetland<br />
near Pigeon Lake, Bayfield County, Wisconsin, U.S.A. A<br />
total of 26 gravid and 110 subadult nematodes were<br />
found in 2 of 4 A. maculatum. Nematodes were fixed in<br />
hot formalin and postfixed in 70% ethanol. Prior to light<br />
microscopic examination, worms were cleared in glycerol<br />
by gradual evaporation from a 5% solution of glycerol in<br />
70% ethanol. Nematodes to be examined with scanning<br />
electron microscopy (SEM) were postfixed in ethanol, dehydrated<br />
in a graded series of ethanol and acetone, and<br />
critical point dried in a Desk II Critical Point Dryer®<br />
(Denton Vacuum, Inc., Moorestown, New Jersey, U.S.A.)<br />
with CO2 as the transition fluid. The specimens were<br />
mounted on stubs, coated with gold, and examined with<br />
a Hitachi 2460N® scanning electron microscope (Hitachi<br />
USA, Mountain View, California, U.S.A.) at an accelerating<br />
voltage of 10-15 kV<br />
Five specimens of R. bermani from S. keyserlingii collected<br />
in Magadanskaya Region, Russia, 10 specimens of<br />
R. tokyoensis from the brown newt Cynops ensicauda<br />
(Hallowell, 1860) collected on Okinawa Island, Japan, 20<br />
specimens of R. ranae from the northern leopard frog<br />
Rana pipiens (Schreber, 1782) collected in Wisconsin,<br />
U.S.A., and 18 specimens of Rhabdias americanus Baker,<br />
1978, from the American toad Bufo americanus Hoibrook,<br />
1836, collected in Wisconsin, U.S.A. were examined<br />
by light microscopy and measured after being<br />
cleared as above. All measurements are given in micrometers<br />
unless otherwise stated. Measurements are given<br />
for the holotype followed by minimum and maximum<br />
measurements of paratypes in parentheses.
KUZMIN ET AL.—RHABDIAS AMBYSTOMAE SP. N. 229<br />
Table 1. Measurements taken from gravid Rhabdias ambystomae sp. n. (type series; n = 18) (measurements<br />
in micrometers unless otherwise noted).<br />
Character<br />
Body length (mm)<br />
Body width<br />
Buccal capsule depth<br />
Buccal capsule width<br />
Width of esophagus, anterior end<br />
Width of esophagus, muscular region<br />
Minimum width of esophagus, glandular region<br />
Esophageal bulb width<br />
Distance, anterior end of esophagus to nerve ring<br />
Distance, anterior end of esophagus to nerve ring (as % of esophagus length)<br />
Esophagus length<br />
Esophagus length (as % of body length)<br />
Distance from anterior end to vulva (mm)<br />
Distance from anterior end to vulva (as % of body length)<br />
Tail length<br />
Tail length (as % of body length)<br />
Description<br />
Results<br />
Rhabdias ambystomae sp. n.<br />
(Figs. 1-15)<br />
Because both gravid and subadult nematodes<br />
were found in the same host specimens, each of<br />
these stages is described separately.<br />
GRAVID SPECIMENS (Table 1): Body elongated<br />
12.6 (6.8-13.0) mm long, 350 (210-430) wide.<br />
Anterior end rounded, posterior end tapered. Body<br />
cuticle swollen, especially on anterior and posterior<br />
thirds of body. Irregularly arranged transverse<br />
folds formed by cuticular surface. Round oral<br />
opening surrounded by 6 small lips, each bearing<br />
1 elongated conical inner papilla and 2 minute outer<br />
papillae. Inner papillae directed toward oral<br />
opening. Flat cuticular ring separating each lip<br />
from edge of oral opening. Buccal capsule cuplike<br />
in lateral view, round in apical view. Buccal<br />
capsule depth 15 (12-15), width 17 (15-17).<br />
Esophagus club-shaped, 560 (450-590) in length,<br />
with short anterior muscular portion and long posterior<br />
glandular portion. Nerve ring at level of border<br />
between muscular and glandular regions of<br />
esophagus, 160 (130-180) from esophagus anterior<br />
end. Large optically dense hypodermal cells<br />
prominent along glandular region of esophagus.<br />
Excretory glands indistinct, excretory duct short.<br />
Two large anterior coelomocytes situated between<br />
posterior end of esophagus and loop of anterior<br />
genital limb. Intestine thick, filled with brown or<br />
Holotype<br />
12.6<br />
350<br />
15<br />
17<br />
45<br />
50<br />
57<br />
90<br />
160<br />
28.6<br />
560<br />
4.5<br />
7<br />
56<br />
210<br />
1.7<br />
Paratypes<br />
(mean [min.— max.])<br />
10.5 (6.8-13.0)<br />
342 (210-430)<br />
14 (12-15)<br />
17 (15-17)<br />
43 (40-45)<br />
54 (45-67)<br />
59 (42-67)<br />
85 (65-100)<br />
159 (130-180)<br />
28.9 (22.8-34.6)<br />
546 (450-590)<br />
5.4 (4.1-7.4)<br />
5.8 (3.8-7.4)<br />
55 (47.4-58.2)<br />
236 (190-300)<br />
2.3 (1.6-3.1)<br />
black contents. Intestinal walls thinner in posterior<br />
than in anterior region of body. Muscular sphincter<br />
between intestine and rectum present. Rectum<br />
lined with thick cuticle. Tail wide, conical, 210<br />
(190-300) in length. Vulva usually postequatorial<br />
with indistinct lips. Genital system amphidelphic.<br />
Ovaries straight or slightly twisted, lying along intestine.<br />
Proximal regions of both ovaries overlap<br />
level of vulva. Both limbs of genital system bend<br />
backward at level of oviducts. Anterior oviduct occasionally<br />
forms 2 loops as it bends. Seminal receptacles<br />
short, thick walled. Uteri wide, thin<br />
walled, filled with numerous eggs. Egg size 112-<br />
130 X 55-65.<br />
SUBADULT SPECIMENS (Table 2): Body length<br />
4.15 (3.3-4.8) mm, width 111 (100-120). Anterior<br />
end rounded, posterior end tapered. Body cuticle<br />
thin and smooth, slightly swollen at anterior and<br />
posterior ends. Head structures similar to those in<br />
gravid worms. Lateral lips situated farther from<br />
oral opening than submedian lips. Buccal capsule<br />
and esophagus shapes similar to those in adults.<br />
Buccal capsule 12 (10-12) deep, 17 (15-17) wide.<br />
Esophagus 432 (400-460) long. Two elongated<br />
narrow excretory glands stretch from posterior<br />
edge of nerve ring to anterior end of intestine. Pair<br />
of coelomocytes situated subventrally, close to anterior<br />
limb of genital system. Intestine thick, reddish.<br />
Rectum sclerotized. Tail elongated, 171<br />
(150-190) in length. Bulbous projection of body<br />
wall slightly posteriad to anal opening. Vulva postequatorial.<br />
Vulva lips indistinct. Genital system<br />
Copyright © 2011, The Helminthological Society of Washington
230 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Figures 1-4. Rhabdias ainbystomae sp. n., adult. 1. Anterior end. 2. Head end, lateral view. 3. Cephalic<br />
extremity, apical view. 4. Posterior end. 1, 2, 4, holotype; 3, paratype.<br />
Copyright © 2011, The Helminthological Society of Washington
KUZMIN ET AL.—RHABD1AS AMBYSTOMAE SP. N. 231<br />
Table 2. Measurements of Rhabdias ambystomae sp. n. subadult specimens (n = 15) (measurement in<br />
micrometers unless otherwise noted).<br />
Character<br />
Body length (mm)<br />
Body width<br />
Buccal capsule depth<br />
Buccal capsule width<br />
Width of esophagus, anterior end<br />
Width of esophagus, muscular region<br />
Minimum width of esophagus, glandular region<br />
Esophageal bulb width<br />
Distance, anterior end of esophagus to nerve ring<br />
Distance, anterior end of esophagus to nerve ring (as % of esophagus length)<br />
Esophagus length<br />
Esophagus length (as % of body length)<br />
Distance from anterior end to vulva (mm)<br />
Distance from anterior end to vulva (as % of body length)<br />
Tail length<br />
Tail length (as % of body length)<br />
completely developed, but eggs absent. Proximal<br />
regions of gonads overlap level of vulva. Each gonad<br />
forms a single loop as it bends. Uteri narrow,<br />
lacking eggs.<br />
Taxonomic summary<br />
TYPE HOST: Spotted salamander Arnbystoma<br />
maculatum (Shaw, 1802).<br />
TYPE LOCALITY: Roadside wetland near Pigeon<br />
Lake, Bayfield County, Wisconsin, U.S.A.;<br />
46°20'84"N, 91°20'58"W.<br />
SITES OF INFECTION: Lungs, body cavity.<br />
TYPE SPECIMENS: The type series consists of<br />
the gravid specimens only. Holotype: U.S. National<br />
Parasite Collection, Beltsville, Maryland,<br />
U.S.A., USNPC 90869. Paratypes: USNPC<br />
90870 (9 specimens); Department of <strong>Parasitology</strong>,<br />
Institute of Zoology, Kiev, Ukraine, Vial N<br />
847 (8 specimens).<br />
ETYMOLOGY: The new species is named in<br />
reference to the generic name of its type host.<br />
PREVALENCE AND INTENSITY: Two of 4 specimens<br />
of spotted salamander; 20-116 specimens<br />
of R. ambystomae (5-21 adult nematodes in<br />
lungs and 15-95 subadults in body cavity).<br />
Remarks and Discussion<br />
Rhabdias ambystomae sp. n. is most similar<br />
morphologically to R. bermani and R. tokyoensis,<br />
the only other species of Rhabdias described<br />
from salamanders. The 3 species are similar in<br />
body size and shape and in egg size (Table 3).<br />
Rhabdias ambystomae sp. n. differs from R. ber-<br />
Mean<br />
4.2<br />
11 1<br />
12<br />
17<br />
37<br />
39<br />
42<br />
57<br />
149<br />
34.4<br />
432<br />
10.5<br />
2.4<br />
57.4<br />
171<br />
4.1<br />
Minimum<br />
3.3<br />
100<br />
10<br />
15<br />
35<br />
35<br />
37<br />
50<br />
110<br />
26.8<br />
400<br />
8.8<br />
1.9<br />
54.4<br />
150<br />
3.5<br />
Maximum<br />
4.9<br />
120<br />
12<br />
17<br />
40<br />
42<br />
47<br />
60<br />
170<br />
37.8<br />
460<br />
12.0<br />
2.8<br />
61.8<br />
190<br />
5.0<br />
mani in the absence of a lancet-like cuticular<br />
swelling of the posterior extremity characteristic<br />
of the latter species. In R. bermani, the circumoral<br />
lips are arranged in 2 lateral groups; in each<br />
group, the lateral lip is situated closer to the oral<br />
opening than are the submedian lips (Rausch et<br />
al., 1984). In contrast, the lateral lips of R. ambystomae<br />
sp. n. are situated farther from the oral<br />
opening than are the submedian lips, a character<br />
more prominent in subadult worms than in<br />
adults (Figs. 3, 7). Additionally, the vulva of R.<br />
ambystomae sp. n. is usually postequatorial,<br />
whereas it is equatorial in R. bermani (Rausch<br />
et al., 1984). The most apparent differences between<br />
R. ambystomae sp. n. and R. tokyoensis<br />
are the markedly smaller buccal capsule and narrower<br />
esophagus of R. ambystomae (Table 3).<br />
Rhabdias ambystomae sp. n. can be distinguished<br />
readily from 2 Rhabdias species common<br />
in North American amphibians, R. atnericanus<br />
and R. ranae, by the absence of lateral<br />
pseudolabia on the cephalic end. The presence<br />
and shape of pseudolabia in R. americanus and<br />
R. ranae were well documented by Baker (1978)<br />
and confirmed by examination of specimens collected<br />
as part of the present study. In addition,<br />
R. americanus has a more elongated tail (cf.<br />
Baker, 1978, fig. 3) compared with that of R.<br />
ambystomae sp. n.<br />
Parasitic nematodes of the genus Rhabdias<br />
have been reported in several species of salamanders<br />
from the midwestern U.S.A. Price and<br />
St. John (1980) reported finding an undeter-<br />
Copyright © 2011, The Helminthological Society of Washington
232 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Figures 5-9. Rhabdias ambystomae sp. n., subadult. 5. Anterior end. 6. Head end, lateral view. 7.<br />
Cephalic extremity, apical view. 8. Posterior end. 9. Anterior loop of genital system and coelomocytes.<br />
mined species of Rhabdias in the smallmouth ( = subadult) nematodes were found in the body<br />
salamander Ambystoma texanum (Matthes, cavity. Bolek and Coggins (1998) found unde-<br />
1855) from Illinois. Adult specimens were found termined subadult Rhabdias in the body cavity<br />
in the lungs of the host, whereas "larval" of the lungless red-backed salamander Pletho-<br />
Copyright © 2011, The Helminthological Society of Washington
KUZMIN ET AL.—RHABDIAS AMBYSTOMAE SP. N. 233<br />
Figures 10-15. External morphology of Rhabdias ambystomae sp. n. 10. Cephalic extremity of adult.<br />
11. Cephalic extremity of subadult; note amphid marked with an arrow. 12. Vulva of adult. 13. Vulva of<br />
subadult; note obliteration of the female genital opening at this stage. 14. Anus of adult. 15. Anus of<br />
subadult. Scale bars =<br />
Copyright © 2011, The Helminthological Society of Washington
234 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
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Copyright © 2011, The Helminthological Society of Washington<br />
rfon cinereus (Green, 1818) from collecting sites<br />
within a few kilometers of where R. ambystomae<br />
sp. n. was collected as part of the present study<br />
(J. R. Coggins, University of Wisconsin, Milwaukee,<br />
personal communication). Oddly<br />
enough, no Rhabdias were found in A. maculaturn<br />
collected by Bolek and Coggins (1998)<br />
from the same area. This information suggests<br />
that R. ambystomae sp. n. may also parasitize P.<br />
cinereus in northern Wisconsin but cannot reach<br />
maturity in this lungless amphibian. Further examinations<br />
will be necessary for confirmation.<br />
Other authors identified lung nematodes collected<br />
from salamanders in Wisconsin and Michigan<br />
as R. ranae (Coggins and Sajdak, 1982;<br />
Muzzall and Schinderle, 1992). This species has<br />
been recorded in a number of North American<br />
anurans, most of which belong to the family<br />
Ranidae (Baker, 1978; Yoder and Coggins,<br />
1996; Bursey and DeWolf, 1998). In our opinion,<br />
the determination of the material from salamanders<br />
as R. ranae is questionable because of<br />
the high level of host specificity demonstrated<br />
by most representatives of Rhabdias; members<br />
of this genus have never been found to parasitize<br />
hosts from more than a single order (Rausch et<br />
al., 1984). Similarly, the report by Goldberg et<br />
al. (1998) of a frog parasite, Rhabdias joaquinensis<br />
Ingles, 1935, from salamanders in California<br />
may also represent a misidentification and<br />
is in need of confirmation.<br />
Rhabdias ambystomae sp. n. is the first species<br />
of this genus described from North American<br />
salamanders. The rich salamander fauna of<br />
North America and the strict specificity of rhabdiasids<br />
to their hosts indicate that further studies<br />
of material from the field or museum collections<br />
may reveal the presence of more species of<br />
Rhabdias unique to the salamanders of the New<br />
World.<br />
Acknowledgments<br />
We thank Dr. Gennadiy Atrashkevich, who<br />
kindly provided several specimens of 5. keyserlingii<br />
for our investigation, and Dr. Hideo Hasegawa<br />
for the loan of R. tokyoensis. We are<br />
grateful to Dr. Robert Wise for assistance with<br />
SEM. Collection of amphibians in Wisconsin<br />
was conducted under a permit provided by the<br />
Wisconsin Department of Natural Resources.<br />
This research was supported by a grant from the<br />
Vander Putten International Fund of the University<br />
of Wisconsin Oshkosh.
Literature Cited<br />
Baker, M. R. 1978. Morphology and taxonomy of<br />
Rhabdias spp. (Nematoda: Rhabdiasidae) from<br />
reptiles and amphibians of southern Ontario. Canadian<br />
Journal of Zoology 56:2127-2141.<br />
Bolek, M. G., and J. R. Coggins. 1998. Helminth<br />
parasites of the spotted salamander Ambystoma<br />
maculatum and red-backed salamander Plethodon<br />
c. cinereus from northwestern Wisconsin. Journal<br />
of the Helminthological Society of Washington<br />
65:98-102.<br />
Bursey, C. R., and W. R. DeWolf. 1998. Helminths<br />
of the frogs, Rana catesbeiana, Rana clamitans,<br />
and Rana palustris, from Coshocton County,<br />
Ohio. Ohio Journal of Science 98:28-29.<br />
Coggins, J. R., and R. A. Sajdak. 1982. A survey of<br />
helminth parasites in the salamanders and certain<br />
anurans from Wisconsin. Proceedings of the Helminthological<br />
Society of Washington 49:99-102.<br />
Dyer, W. G., and S. B. Peck. 1975. Gastrointestinal<br />
parasites of the cave salamander, Eurycea lucifuga<br />
Rafinesque, from the south-eastern United <strong>State</strong>s.<br />
Canadian Journal of Zoology 53:52-54.<br />
Goldberg, S. R., C. R. Bursey, and H. Cheam. 1998.<br />
Composition and structure of helminth communities<br />
of the salamanders, Aneides lugubrls, Batrachoseps<br />
nigriventris, Ensatina eschscholtzii<br />
(Plethodontidae), and Taricha torosa (Salamandridae)<br />
from California. Journal of <strong>Parasitology</strong><br />
84:248-251.<br />
KUZMIN ET AL.—RHABDIAS AMBYSTOMAE SP. N. 235<br />
Lehmann, D. L. 1954. Some helminths of west coast<br />
urodels. Journal of <strong>Parasitology</strong> 40:231.<br />
Muzzall, P. M., and D. B. Schinderle. 1992. Helminths<br />
of the salamanders Ambystoma t. tigrinum<br />
and Ambystoma laterale (Caudata, Ambystomatidae)<br />
from southern Michigan. Proceedings of the<br />
Helminthological Society of Washington 59:201-<br />
205.<br />
Price, R. L., and T. St. John. 1980. Helminth parasites<br />
of the small-mouth salamander, Ambystoma<br />
texamim Matthes, 1855, from William County, Illinois.<br />
Proceedings of the Helminthological Society<br />
of Washington 47:273-274.<br />
Rausch, R. L., V. R. Rausch, and G. I. Atrashkevich.<br />
1984. Rhabdias bermani sp. n. (Nematoda,<br />
Rhabdiasidae) from the Siberian salamander (Hynobius<br />
keyseiiingi) from the north-east of Asia.<br />
Zoologicheskiy Zhurnal 63:1297-1303. (In Russian.)<br />
Wilkie, J. S. 1930. Some parasitic nematodes from<br />
Japanese Amphibia. Annals and Magazine of Natural<br />
History 6:606-614.<br />
Yamaguti, S. 1935. Studies on the helminth fauna of<br />
Japan. Part 10. Amphibian nematodes. Japanese<br />
Journal of Zoology 6:387-392.<br />
Yoder, H. R., and J. R. Coggins. 1996. Helminth<br />
communities in the northern spring peeper, Pseudacris<br />
crucifer Wied, and the wood frog, Rana<br />
sylvatica Le Conte, from southeastern Wisconsin.<br />
Journal of the Helminthological Society of Washington<br />
63:211-214.<br />
Copyright © 2011, The Helminthological Society of Washington
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 236-241<br />
Supplemental Diagnosis of Myxobolus gibbosus (Myxozoa), with a<br />
Taxonomic Review of Myxobolids from Lepomis gibbosus<br />
(Centrarchidae) in North America<br />
DAVID K. CONE<br />
Department of Biology, Saint Mary's University, Halifax, Nova Scotia, Canada B3H 3C3<br />
(e-mail: david.cone@stmarys.ca)<br />
ABSTRACT: Myxobolus gibbosus Herrick, 1941 (Myxosporea) is reported from the connective tissue of gills of<br />
Lepomis gibbosus (Centrarchidae) in Algonquin Park, Ontario, Canada. The new material (formalin-preserved)<br />
is used to supplement the original taxonomic diagnosis of nearly 60 yr ago. Spores are round to oval in valvular<br />
view, 11-14 jjim long and 10-11 jxm wide, with a distinctly blunt capsular region. The polar capsules are<br />
relatively large for the size of the spore, measuring 6-7 jxm long and 3.5-4.0 (Jim wide, and aligned almost<br />
parallel to each other. There are 8-12 loose filament coils lying up to 45° to the long axis of the capsule. The<br />
taxonomy of species of Myxobolus described or reported from L. gibbosus in North America is examined, and<br />
the following are considered to be valid taxa: Myxobolus dechtiari Cone and Anderson, 1977; M. gibbosus;<br />
Myxobolus magnasphcrus Cone and Anderson, 1977; Myxobolus osburni Herrick, 1936; Myxobolus paralintoni<br />
Li and Desser, 1985; and Myxobolus uvidiferus Cone and Anderson, 1977. <strong>Comparative</strong> photographs of spores<br />
accompany differential diagnoses of the 6 species. Myxobolus gibbosus Li and Desser, 1985, and Myxobolus Hi<br />
Desser, 1993, are junior synonyms of M. uvidiferus. Myxobolus lepomicus Li and Desser, 1985, is considered a<br />
species inquirendae, and the reports of Myxobolus cyprinicola Reuss, 1906, and Myxobolus poecilichthidis<br />
Fantham, Porter, and Richardson, 1939, from L. gibbosus are considered misidentifications.<br />
KEY WORDS: Myxobolus gibbosus, Myxosporea, redescription, differential diagnoses, pumpkinseed sunfish,<br />
Lepomis gibbosus, Centrarchidae, Algonquin Park, Canada.<br />
Myxobolus gibbosus Herrick, 1941 (Myxozoa)<br />
was described from connective tissue of the<br />
gill arch of pumpkinseed sunfish (Lepomis gibbosus<br />
(Linnaeus, 1758)) from the island region<br />
of western Lake Erie (Herrick, 1941). In subsequent<br />
surveys of myxosporean parasites of<br />
pumpkinseed (Cone and Anderson, 1977a, b; Li<br />
and Desser, 1985; Hayden and Rogers, 1997),<br />
the parasite was not encountered. However, during<br />
a new survey of myxosporeans of fish in<br />
Algonquin Park, a single pseudocyst of M. gibbosus<br />
was discovered. This rare find enabled the<br />
author to assess information provided in the<br />
original species description and to critically<br />
compare the parasite with other species of the<br />
genus reported from pumpkinseed. The present<br />
study describes the new material and reviews the<br />
taxonomy of myxobolids from pumpkinseed in<br />
North America.<br />
Materials and Methods<br />
Nine pumpkinseed (6-8.9 cm in total length) were<br />
collected in baited trapnets set 20 June 1994 and 21<br />
June 1995 in the shallows of Lake Sasajewan<br />
(45°35'N; 78°30'W), Algonquin Park, Ontario, Canada.<br />
The fish were pithed and necropsied. All body organs<br />
and tissues were examined microscopically for<br />
myxosporean pseudocysts, and, when found, they were<br />
236<br />
fixed in 10% buffered formalin. Fixed pseudocysts<br />
were punctured and the spore contents stabilized in<br />
temporary mounts prepared with 1% agar (Lom,<br />
1969). Spores were photographed with interference<br />
contrast optics. Enlarged photographic prints of individual<br />
spores were used to determine spore dimensions.<br />
Descriptive terminology follows Lom and Dykova<br />
(1992). Measurements are presented in micrometers.<br />
The sample of M. gibbosus was compared with<br />
other species of Myxobolus in the author's collection,<br />
namely Myxobolus dechtiari Cone and Anderson,<br />
1977; Myxobolus magnaspherus Cone and Anderson,<br />
1977; Myxobolus osburni Herrick, 1936; Myxobolus<br />
paralintoni Li and Desser, 1985; and Myxobolus uvitliferus<br />
Cone and Anderson, 1977. Syntype slides of M.<br />
gibbosus (NMCICP 1984-0359), Myxobolus lepomicus<br />
(NMCICP 1984-0362), and M. paralintoni (NMCICP<br />
1984-0364) housed in the parasite collection of the Canadian<br />
Museum of Nature were also examined. A photo-voucher<br />
(negative film) is deposited in the United<br />
<strong>State</strong>s National Parasite Collection (USNPC), Beltsville,<br />
Maryland, U.S.A.<br />
Results<br />
Myxobolus gibbosus Herrick, 1941<br />
(Figs. 1 and 2)<br />
Supplementary diagnosis<br />
Copyright © 2011, The Helminthological Society of Washington<br />
Pseudocyst egg-shaped, gray-white and minute<br />
(250 long), embedded in connective tissue<br />
surrounding base of gill arch. Spores round to
Figure 1. Spores of Myxobolus gibbosus. Scale bar = 10 [Jim.<br />
oval in valvular view, with blunt capsular edge.<br />
Spores 11.8 ± 0.9 (11-14, « = 10) long and<br />
10.6 ± 0.5 (10-1 1) wide. Width-to-length ratio<br />
1:1.09 ± 0.08 (1.05-1.3). Polar capsules oval,<br />
6.8 ± 0.3 (6-7) long and 4.0 ± 0.3 (3.5-4) wide,<br />
aligned almost parallel to each other. Polar filaments<br />
in 8-12 loose coils, lying up to 45° to long<br />
axis of capsule. Capsulogenic nuclei prominent,<br />
triangular. Shallow intercapsular appendix evident<br />
in some spores. Sutural ridge thin and<br />
smooth.<br />
Taxonomic summary<br />
HOST: Pumpkinseed sunfish (Lepomis gibbosus)<br />
(Centrarchidae); total length 6.2 cm, 1 +<br />
yr old.<br />
LOCALITY/COLLECTION DATE: Lake Sasajewan,<br />
Algonquin Park, Ontario, Canada (45°35'N;<br />
78°30'W), 20 June 1994.<br />
CONE—DIAGNOSIS OF MYXOBOLUS 237<br />
SITE OF INFECTION: Connective tissue of gill<br />
arch.<br />
PREVALENCE AND INTENSITY OF INFECTION: One<br />
of 9 fish infected with 1 pseudocyst.<br />
SPECIMENS DEPOSITED: Photo-voucher USNPC<br />
No. 091157.00.<br />
Remarks<br />
Myxobolus gibbosus has not been reported in<br />
surveys of myxozoans in L. gibbosus from Algonquin<br />
Park (Cone and Anderson 1977a, b; Li<br />
and Desser, 1985). It was probably not overlooked,<br />
for the parasite has several distinct diagnostic<br />
features. It forms small but obvious<br />
pseudocysts in the connective tissue of the gill<br />
arch and produces round spores with a blunt<br />
capsular end. The polar capsules are relatively<br />
large, the length being about half the length of<br />
the spore, and they are arranged almost parallel<br />
Copyright © 2011, The Helminthological Society of Washington
238 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Table 1. Comparison of pertinent taxonomic information about Myxobolus gibbosus reported in the original<br />
species description and that observed in the present study.<br />
Host<br />
Locality<br />
Tissue site<br />
Pseudocyst<br />
Spore length<br />
Spore width<br />
Spore thickness<br />
Polar capsule length<br />
Polar capsule width<br />
Polar filament coils<br />
Herrick (1941)*<br />
Lepomis gibbosus<br />
Lake Erie<br />
Connective tissue of gill<br />
Round, 0.75 mm<br />
10.6-12.3<br />
9.8-12.3<br />
6.5-8.2<br />
5.7-7.4<br />
3.3-4.1<br />
8-12<br />
* Based on fresh material in a hanging drop preparation.<br />
t Based on formalin-fixed material in asar wet mounts.<br />
to each other. It appears then that M. gibbosus<br />
is simply rare in this region. The dimensions of<br />
the preserved spores found in the present study<br />
are similar to those described by Herrick (1941)<br />
from fresh material (Table 1). It should be noted<br />
that dimensions of fixed spores are often smaller<br />
than those of fresh spores because shrinkage can<br />
take place during fixation. This means that fresh<br />
spores of M. gibbosus in Algonquin Park may<br />
be slightly larger than those described originally<br />
by Herrick (1941).<br />
Spores of other species of Myxobolus (M. dechtiari,<br />
M. magnaspherus, M. osburni, M. paralintoni,<br />
and M. uvuliferus) from L. gibbosus are<br />
presented for comparative purposes (Figs. 3—7).<br />
Each species has a distinct spore shape and specific<br />
tissue site in which it develops and is readily<br />
identified by these indicators. Myxobolus<br />
paralintoni (Fig. 4) has oval spores in frontal<br />
view and develops in the bulbus arteriosus of the<br />
heart (Hayden and Rogers, 1997; Cone and<br />
Overstreet, 1998). Myxobolus dechtiari (Fig. 5)<br />
has spores that are broadly pyriform in frontal<br />
view and develops in gill tissue (Cone and Anderson,<br />
1977a). Myxobolus uvuliferus has slightly<br />
compressed spores in frontal view usually<br />
with the width greater than length, often has polar<br />
capsules dissimilar in the length, and develops<br />
in the connective tissue capsule surrounding<br />
the metacercaria of Uvulifer ambloplites<br />
(Hughes, 1927) Dubois, 1938 (Cone and Anderson,<br />
1977a). Myxobolus osburni has round<br />
spores in frontal view and develops in the exocrine<br />
tissue of the pancreas (Cone and Anderson,<br />
1977a). Myxobolus magnaspherus has round<br />
spores in frontal view that are huge, often 20<br />
Copyright © 2011, The Helminthological Society of Washington<br />
Present studyt<br />
Lepomis gibbosus<br />
Lake Sasajewan<br />
Connective tissue of gill<br />
Round, 0.25 mm<br />
11-14<br />
10-11<br />
—<br />
6-7<br />
3.5-4<br />
8-11<br />
|xm in diameter, and develops in connective tissue<br />
of the body, including the peritoneum (Cone<br />
and Anderson, 1977a).<br />
Taxonomic Key to the Species of Myxobolus<br />
Infecting Pumpkinseed Sunfish<br />
la. Spore length more than 16 jjim<br />
M. magnaspherus (Fig. 8)<br />
Ib. Spore length less than 16 |xm 2<br />
2a. Polar capsules aligned more or less parallel -<br />
M. gibbosus (Fig. 9)<br />
2b. Polar capsules converged anteriorly 3<br />
3a. Spore circular in frontal view M. osburni (Fig. 10)<br />
3b. Spore not circular in frontal view 4<br />
4a. Spore width greater than spore length<br />
M. uvuliferus (Fig. 11)<br />
4b. Spore width less than spore length 5<br />
5a. Spore oval in frontal view M. paralintoni (Fig. 12)<br />
5b. Spore broadly pyriform in frontal view<br />
M. dechtiari (Fig. 13)<br />
Discussion<br />
Ten species of Myxobolus Butschli, 1882<br />
(Myxosporea) have been reported from L. gibbosus<br />
in North America (Herrick, 1936, 1941;<br />
Cone and Anderson, 1977a, b; Ingram and<br />
Mitchell, 1982; Li and Desser, 1985; Desser,<br />
1993; Cone and Overstreet, 1998). The author<br />
has necropsied L. gibbosus from Algonquin Park<br />
and from Lake Erie and has to date encountered<br />
6 of the 10 species, namely M. dechtiari, M.<br />
gibbosus, M. magnaspherus, M. osburni, M.<br />
paralintoni, and M. uvuliferus.<br />
The reports of Myxobolus cyprinicola Reuss,<br />
1906, and Myxobolus poecilichthidis Fantham,<br />
Porter, and Richardson, 1939, from the brain and<br />
heart and from the gills, respectively, of L. gib-
CONE—DIAGNOSIS OF MYXOBOLUS 239<br />
Figures 2-7. Photographs of spores in frontal view of species of Myxobolus known to parasitize Lepomis<br />
gibbosus in North America. 2. Myxobolus gibbosus (formalin-preserved). 3. Myxobolus paralintoni (formalin-preserved).<br />
4. Myxobolus dechtiari (formalin-preserved). 5. Myxobolus uvuliferus (formalin-preserved).<br />
6. Myxobolus osburni (formalin-preserved). 7. Myxobolus magnaspherus (fresh spore). Scale bar<br />
= 10 jjim and applies to all figures.<br />
bosus in Algonquin Park (Li and Desser, 1985)<br />
are considered misidentifications. Both of these<br />
species have similarities in tissue site and spore<br />
morphology to M. dechtiari and M. paralintoni<br />
and could have easily been confused with them.<br />
Li and Desser (1985) were apparently unaware<br />
of the study by Cone and Anderson (1977a) in<br />
nearby Ryan Lake.<br />
Copyright © 2011, The Helminthological Society of Washington
240 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
8<br />
12<br />
Figures 8-13. Drawings of spores in frontal view of species of Myxobolus known to parasitize Lepomis<br />
gibbosus in North America. 8. Myxobolus gibbosus. 9. Myxobolus paralintoni. 10. Myxobolus dechtiari. 11.<br />
Myxobolus uvuliferus. 12. Myxobolus osburni. 13. Myxobolus magnaspherus. Scale bar = 5 (Jim and applies<br />
to all figures.<br />
The type material of M. lepomicus Li and<br />
Desser, 1985, described from a variety of organs<br />
of L. gibbosus, has deteriorated, and spores are<br />
not to be found on the slide. The species description<br />
includes a schematic drawing of the<br />
spore. Until additional samples are obtained the<br />
species is considered a species inquirendae.<br />
Myxobolus gibbosus Li and Desser, 1985, is a<br />
homonym of M. gibbosus Herrick, 1941. Desser<br />
(1993) proposed Myxobolus Hi as a nomen novum<br />
to replace M. gibbosus Li and Desser, 1985.<br />
However, Landsberg and Lom (1991) considered<br />
M. gibbosus Li and Desser, 1985, to be a<br />
junior synonym of M. uvuliferus, and thus both<br />
M. gibbosus and M. Hi become junior synonyms<br />
of M. uvuliferus. The report by Hoffman (1998)<br />
that M. gibbosus Li and Desser, 1985, is a junior<br />
synonym of M. osburni cannot be supported on<br />
the basis of spore shape.<br />
The 6 confirmed species of Myxobolus mentioned<br />
above are known to parasitize L. gibbosus<br />
or related centrarchid fishes in North America.<br />
Myxobolus magnaspherus and M. paralintoni<br />
have been found in redear sunfish (Lepomis microlophus<br />
(Giinther, 1859)) in Mississippi,<br />
U.S.A. (D. K. Cone, Saint Mary's University,<br />
Copyright © 2011, The Helminthological Society of Washington<br />
and R. M. Overstreet, Gulf Coast Research Laboratory,<br />
unpublished data) and redbreast sunfish<br />
(Lepomis auritus (Linnaeus, 1758)) in Maryland,<br />
U.S.A. (Hayden and Rogers, 1997), respectively.<br />
Myxobolus osburni has been reported<br />
(Herrick, 1936; Otto and Jahn, 1943) from bluegill<br />
sunfish (Lepomis macrochirus Rafinesque,<br />
1819), smallmouth bass (Micropterus dolomieu<br />
Lacepede, 1802), and black crappie (Pomoxis nigromaculatus<br />
(Lesueur, 1829)). The genus clearly<br />
has undergone a diverse radiation in these<br />
hosts, and it is of ecological interest that all 6<br />
species are found in L. gibbosus in Algonquin<br />
Park and that all occupy distinct and very specific<br />
tissue sites in this host species.<br />
Acknowledgments<br />
The research was funded by a Natural Sciences<br />
and Engineering Research Council of<br />
Canada (NSERC) Research Grant awarded to<br />
the author. Thanks are extended to the staff of<br />
the Harkness Research Laboratory for their help<br />
and hospitality and to Sherwin Desser, University<br />
of Toronto, for providing constructive comments<br />
on a draft of the paper.
Literature Cited<br />
Cone, D. K., and R. C. Anderson. 1977u. Myxosporidan<br />
parasites of pumpkinseed (Lepomis gibbosus<br />
L.) from Ontario. Journal of <strong>Parasitology</strong><br />
63:657-666.<br />
, and . 1977b. Parasites of pumpkinseed<br />
(Lepomis gibbosus L.) from Ryan Lake, Algonquin<br />
Park, Ontario. Canadian Journal of Zoology<br />
55:1410-1423.<br />
-, and R. M. Overstreet. 1998. Species of Myxobolus<br />
(Myxozoa) from the bulbus arteriosus of<br />
centrarchid fishes in North America, with a description<br />
of two new species. Journal of <strong>Parasitology</strong><br />
84:371-374.<br />
Desser, S. S. 1993. Myxobolus Hi nom. nov.: a replacement<br />
for M. gibbosus Li and Desser, 1985 (Myxosporea:<br />
Myxozoa), preoccupied. Canadian Journal<br />
of Zoology 71:1461.<br />
Hayden, K. J., and W. A. Rogers. 1997. Redescription<br />
of Myxobolus paralintoni (Myxosporea: Myxobolidae),<br />
with notes regarding new host and locality.<br />
Journal of <strong>Parasitology</strong> 83:283-286.<br />
Herrick, J. A. 1936. Two new species of Myxobolus<br />
from fishes of Lake Erie. Transactions of the<br />
American Microscopical Society 55:194-198.<br />
. 1941. Some myxosporidian parasites of Lake<br />
Erie fishes. Transactions of the American Microscopical<br />
Society 66:163-170.<br />
CONE—DIAGNOSIS OF MYXOBOLUS 241<br />
Hoffman, G. L. 1998. Parasites of North American<br />
Freshwater Fishes, 2nd ed. Comstock Publishing<br />
Associates, Cornell University Press, Ithaca, New<br />
York, U.S.A., and London, U.K. 539 pp.<br />
Ingram, K. M., and L. G. Mitchell. 1982. Pancreatic<br />
infections of Myxobolus osburni Herrick (Myxozoa:<br />
Myxosporea) in the pumpkinseed, Lepomis<br />
gibbosus (Linnaeus), in Iowa. Journal of Wildlife<br />
Diseases 18:75-80.<br />
Landsberg, J. H., and J. Lorn. 1991. Taxonomy of<br />
the genera of the Myxobolus/Myxosoma group<br />
(Myxobolidae: Myxosporea), current listing of<br />
species and revision of synonyms. Systematic <strong>Parasitology</strong><br />
18:165-186.<br />
Li, L., and S. S. Desser. 1985. The protozoan parasites<br />
of fish from two lakes in Algonquin Park,<br />
Ontario. Canadian Journal of Zoology 63:1846-<br />
1858.<br />
Loin, J. 1969. On a new taxonomic character in<br />
Myxosporidia, as demonstrated in descriptions of<br />
two new species. Folia Parasitologica 16:97-103.<br />
, and I. Dykova. 1992. Protozoan Parasites of<br />
Fishes. Developments in Aquaculture and Fisheries<br />
Science 26. Elsevier, Amsterdam, London,<br />
New York, Tokyo. 315 pp.<br />
Otto, G. R., and T. L. Jahn. 1943. Internal myxosporidian<br />
infections of some fishes of the Okobojii<br />
region. Proceedings of the Iowa Academy of Science<br />
50:323-335.<br />
<strong>2001</strong>-2002 MEETING SCHEDULE OF THE<br />
HELMINTHOLOGICAL SOCIETY OF WASHINGTON<br />
27 October <strong>2001</strong><br />
28 October <strong>2001</strong><br />
28 November <strong>2001</strong><br />
January 2002<br />
March 2002<br />
May 2002<br />
Symposium—<strong>Parasitology</strong> in Science. 1:00 PM. Smithsonian Institution,<br />
Quad Building, (Room 3111), Washington, DC<br />
(Contact Persons: Bill Moser, 202-357-2473 or Dennis Richardson, 203-<br />
582-8607).<br />
Presidential Summit of <strong>Parasitology</strong> Societies. 8:30 AM.<br />
Smithsonian Institution, Quad Building, (Room 3112), Washington, DC<br />
(Contact Persons: Bill Moser, 202-357-2473 or Dennis Richardson, 203-<br />
582-8607).<br />
Anniversary Dinner. 6:30 PM. 94th Aero Squadron, <strong>College</strong> Park, MD<br />
(Contact Person: Bill Moser, 202-357-2473).<br />
Date, time, and place to be announced.<br />
Date, time, and place to be announced.<br />
Date, time, and place to be announced.<br />
Copyright © 2011, The Helminthological Society of Washington
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 242-248<br />
Cuticular Changes in Fergusobiid Nematodes Associated with<br />
Parasitism of Fergusoninid Flies<br />
ROBIN M. GiBLiN-DAVis,1-5 KERRIE A. DAViES,2 DONNA S. WiLLiAMS,3 AND<br />
TED D. CENTER4<br />
1 Ft. Lauderdale Research and Education Center, University of Florida, 3205 <strong>College</strong> Avenue, Davie, Florida<br />
33314-7799, U.S.A. (e-mail: giblin@ufl.edu),<br />
2 Department of Applied and Molecular Ecology, Adelaide University, Glen Osmond, 5064 South Australia<br />
(e-mail: kerrie.davies@adelaide.edu.au),<br />
3 Microbiology and Cell Science, University of Florida, P.O. Box 1 10700, Gainesville, Florida 3261 1-0700,<br />
U.S.A. (e-mail: dswill@ufl.edu), and<br />
4 United <strong>State</strong>s Department of Agriculture-Agricultural Research Service, Invasive Plant Research Laboratory,<br />
3205 <strong>College</strong> Avenue, Davie, Florida 33314-7799, U.S.A. (e-mail: tcenter@ars.usda.gov).<br />
ABSTRACT: In the stylet-bearing nematode Fergusobia sp. (Tylenchida: Neotylenchidae), we hypothesize an<br />
additional separation (apolysis) and loss (ecdysis) of the adult cuticle, without the formation of a new cuticle,<br />
during the transition from the preparasitic to parasitic female. This pattern is in direct contrast to the characteristic<br />
4-molt pattern accepted for most nematodes. Transmission electron microscope comparisons of the cuticle of an<br />
adult parthenogenetic female, male, and preparasitic female from the plant-parasitic phase of the fergusobiid life<br />
cycle revealed a relatively simple cuticle with an epicuticle, amorphous cortical/median zone, and a striated<br />
basal zone that is underlain by a relatively thin epidermis and striated somatic muscles. In contrast, the parasitic<br />
female from the adult fly was without its stylet and cuticle, the epidermis was enlarged, the outer edges of the<br />
epidermis were modified into microvilli, and the somatic muscles and esophagus were degenerate. The apparent<br />
hypertrophy and development of epidermal microvilli greatly expand the surface area of the parasitic female<br />
and presumably increase the nematode's ability to absorb nutrients directly through the epidermis from the host's<br />
hemolymph without cuticular interference.<br />
KEY WORDS: Fergusobia, parasitism, Fergusonina, cuticle, epidermis, TEM, molting, nematode, fly, Myrtaceae,<br />
Australia.<br />
In the only known mutualistic association between<br />
nematodes and insects (Maggenti, 1982),<br />
nematodes of the genus Fergusobia Currie,<br />
1937, together with flies of the genus Fergusonina<br />
Currie, 1937, induce galls in young meristematic<br />
tissues of myrtaceous hosts in Australasia<br />
(Giblin-Davis et al., <strong>2001</strong>). The nematode is apparently<br />
responsible for gall induction (Currie,<br />
1937; R. M. Giblin-Davis, unpublished data),<br />
and the fly for dispersal and sustenance of the<br />
nematode. The female fly deposits its eggs and<br />
juvenile nematode parasites in plant tissue (Currie,<br />
1937). As these nematodes feed, a gall begins<br />
to form, and the nematodes develop into<br />
parthenogenetic females that lay eggs giving rise<br />
to amphimictic male and female nematodes. Inseminated<br />
preparasitic females are infective and<br />
invade mature female third-instar fly larvae. Inside<br />
the fly, the nematodes develop into parasitic<br />
females that deposit eggs in the fly's hemolymph.<br />
Juvenile nematodes that hatch from these<br />
Corresponding author.<br />
242<br />
Copyright © 2011, The Helminthological Society of Washington<br />
eggs move to the oviducts of the adult fly and,<br />
together with the fly's eggs, are deposited into<br />
appropriate plant tissue to begin the next generation.<br />
During dissections of mature third-instar fly<br />
larvae from a variety of myrtaceous hosts<br />
(swamp bloodwood Corymbia ptychocarpa (F.<br />
Mueller, 1859), South Australian blue gum Eucalyptus<br />
leucoxylon F. Mueller, 1855, and broadleaved<br />
paperbark Melaleuca quinquenervia (Cavanilles,<br />
1797) S. T. Blake, 1958), we observed<br />
apparent separation (apolysis) and loss (ecdysis)<br />
of the adult cuticle during transition from the<br />
preparasitic to parasitic female without the formation<br />
of a new cuticle (K. A. Davies, unpublished<br />
data). This assumes that the first molt occurs<br />
in the egg in Fergusobia, as with other Tylenchida,<br />
and that 3 molts occur after emergence<br />
from the egg through to the preparasitic female.<br />
This pattern is surprising because nematodes<br />
characteristically undergo 4 molts in their development<br />
from the juvenile to the adult stage<br />
(Bird and Bird, 1991). We report on the ultra-
G1BLIN-DAVIS ET AL.—CUTICULAR CHANGES IN FERGUSOBIID NEMATODES 243<br />
structure of the cuticle of adults at different<br />
phases of the life cycle of Fergusobia.<br />
Materials and Methods<br />
Multilocular flower bud galls of undescribed species<br />
of Fergusobia and Fergusonina were collected on 9<br />
August 1999 from C. ptychocarpa at the Sherwood<br />
Arboretum in Sherwood, Queensland, Australia<br />
(27°32.06'S; 152°58.39'E). Galls were dissected. Adult<br />
parthenogenetic female and amphimictic male and preparasitic<br />
infective female nematodes present in the<br />
plant tissue were placed separately into Trump's fixative<br />
for transmission electron microscopy or in formalin-aceto-alcohol<br />
fixative (Southey, 1970). Mature<br />
fly larvae (third-instar) and adults were dissected from<br />
the galls in phosphate-buffered saline (pH 7.2). Parasitic<br />
female nematodes were removed from the hemocoel<br />
and placed into Trump's fixative. Specimens<br />
were postfixed in 2% formaldehyde (prepared from<br />
paraformaldehyde), 2% glutaraldehyde in 0.1 M cacodylate<br />
buffer at pH 7.2 for 18 hr at 4°C. After repeated<br />
rinsing in buffer, specimens were postfixed in<br />
2% OsO4 in 0.1 M cacodylate buffer at pH 7.2 for 3<br />
days at 4°C. Nematodes were rinsed in water, fixed<br />
with 1 % aqueous uranyl acetate, dehydrated through<br />
100% ethanol into 100% acetone, and infiltrated with<br />
Spurr's epoxy resin. Blocks were sectioned on an<br />
RMC® ultramicrotome. Sections were poststained with<br />
5% aqueous uranyl acetate and lead citrate before<br />
viewing on a Zeiss EM 10® transmission electron microscope<br />
at 80 kV.<br />
Results<br />
Examination of the cuticle of an adult parthenogenetic<br />
female and a male nematode revealed<br />
a relatively simple cuticle with a striated basal<br />
zone, an amorphous cortical/median zone, and a<br />
distinct epicuticle (Figs. 1, 2). It is underlain by<br />
relatively thin epidermis that covers the striated<br />
somatic muscles.<br />
Comparisons of the preparasitic female nematode<br />
from the plant gall and the parasitic female<br />
nematode from the adult fly show dramatic differences<br />
(Figs. 3-7). The preparasitic female has<br />
cuticle, epidermis, and muscles similar to those<br />
described for the male and parthenogenetic female<br />
from the plant host (Figs. 3, 4). However,<br />
the cuticle appears thinner (200-250 nm vs.<br />
450-550 nm for the parthenogenetic female and<br />
630—<strong>68</strong>0 nm for the male). The parasitic form<br />
of the nematode from the adult fly has no cuticle.<br />
The epidermis is greatly enlarged, and the<br />
outer edge of the epidermis appears to be modified<br />
into microvilli (Figs. 5—7). The somatic<br />
muscles appear degenerated (Fig. 6).<br />
During the transition from the preparasitic to<br />
the parasitic female nematode in the larval fly,<br />
the stylet is lost and the esophagus and intestine<br />
appear to degenerate. In a parasitic female from<br />
a fly larva, the remnant of the adult epicuticle<br />
was present (Fig. 5), but it was not present in<br />
the parasitic female from an adult fly (Figs. 6,<br />
7). The apparent hypertrophy and development<br />
of epidermal microvilli greatly expand the surface<br />
area of the parasitic female and presumably<br />
increase the nematode's ability to absorb nutrients<br />
directly through its epidermis from the<br />
host's hemolymph without cuticular interference.<br />
Interestingly, the cuticle represents a form<br />
of protection against insect host defense mechanisms.<br />
However, these mechanisms may be<br />
modified or lacking in the female larva, pupa,<br />
and adult fly in this mutualistic association.<br />
Whether there is a strong defense system in male<br />
flies to prevent parasitism by Fergusobia or the<br />
nematodes fail to penetrate the male fly larvae<br />
is not known.<br />
Discussion<br />
Riding (1970) reported that microvilli were<br />
present on the outside of the parasitic female<br />
stage of Howardula husseyi Richardson, Hesling,<br />
and Riding, 1977 (=Bradynema sp.) (Allantonematidae),<br />
a tylenchid parasite of the<br />
phorid fly, Megaselia halterata Wood, 1910. A<br />
cuticle was not observed in this stage of the<br />
nematode, suggesting that the microvilli were of<br />
epidermal origin and that there could have been<br />
an additional apolysis and ecdysis without cuticular<br />
replacement, as appears to occur in Fergusobia.<br />
The epidermis in this nematode was hypertrophied.<br />
In addition, the stylet and esophagus<br />
are not present in this form of H. husseyi<br />
(Poinar, 1979). Subbotin et al. (1994) reported<br />
that entomoparasitic females of Wachekitylenchus<br />
bovieni (Wachek, 1955) Slobodyanyuk,<br />
1986 (Parasitylenchidae), and Bradynema rigidum<br />
(von Siebold, 1836) zur Strassen, 1892 (Allantonematidae),<br />
had similar body wall morphology<br />
to H. husseyi. Entomoparasitic females<br />
of the tylenchid Skarbilovinema laumondi Chizhov<br />
and Zakharenkova, 1991 (lotonchiidae),<br />
exhibited a body wall composed of a "spongy"<br />
layer of the epidermis formed by interwoven<br />
and fused microvilli without a cuticle (Subbotin<br />
et al., 1993).<br />
In contrast, the epicuticle is apparently retained<br />
by the entomoparasitic amphimictic female<br />
of Paraiotonchium nicholasi Slobodyanyuk,<br />
1975 (=Heterotylenchus sp.) (lotonchiidae)<br />
(Nicholas, 1972). Ultrastructural differences<br />
Copyright © 2011, The Helminthological Society of Washington
244 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
629 nm<br />
;./::•" •':6,'-, -,:•••::•.,;-;<br />
mkdM \•<br />
Figures 1, 2. Longitudinal sections of the cuticle of Fergusobia sp. ex Corymbia ptychocarpa from galled<br />
flower buds. 1. Adult parthenogenetic female. 2. Adult male. BS = basal striations; C/M = cortical/median<br />
zone; E = epicuticle.<br />
were observed between the amphimictic female,<br />
parthenogenetic female, and amphimictic-phase<br />
juvenile of P. nicholasi from the body cavity of<br />
the Australian bush fly Musca vetustissima<br />
Copyright © 2011, The Helminthological Society of Washington<br />
Walker, 1857 (Nicholas, 1972). Fourth-stage juvenile<br />
(J4) nematodes are deposited into cow<br />
dung where they mate, "metamorphose" into<br />
the infective form, penetrate the fly larva, and
-<br />
GIBLIN-DAVIS ET AL.—CUTICULAR CHANGES IN FERGUSOBIID NEMATODES 245<br />
..<br />
'•••':«•'<br />
-<br />
244 nm<br />
•'?"... -<br />
• .<br />
Figures 3, 4. Longitudinal sections of the cuticle of a preparasitic adult female of Fergusobia sp. ex<br />
Corymbia ptychocarpa from galled flower buds. 3. Close-up showing epidermal folds. 4. Different section.<br />
BS = basal striations; C/M = cortical/median zone; E = epicuticle; Ep = epidermis; M = muscle.<br />
Copyright © 2011, The Helminthological Society of Washington
246 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
CR Mv<br />
V/"7f:.:v r^25"/ '••'•:%6&^!£^^±-^<br />
- > V - : : . : ;•- '
GIBLIN-DAVIS ET AL.—CUTICULAR CHANGES IN FERGUSOBIID NEMATODES 247<br />
harenkova and Chizhov, 1991, and Allantonema<br />
rnirabile Leuckart, 1884 (Allantonematidae),<br />
were similar to those of P. nicholasi, being composed<br />
of a hypertrophied epidermis with microvilli<br />
that was covered by a cuticle-like layer<br />
(Subbotin et al., 1994).<br />
Insect hemolymph characteristically has high<br />
levels of amino acids, trehalose, other nonamino<br />
organic acids, and salts (Chapman, 1972), making<br />
it a nutrient-rich environment for parasites<br />
that can overcome innate host defense mechanisms.<br />
Insect-parasitic tylenchid nematodes have<br />
adapted to the challenges of obtaining nutrition<br />
from a living insect host in a variety of ways,<br />
including acquisition per os (through the<br />
mouth), through a modified or absent cuticle, or<br />
through prolapsis and modification of the uterus<br />
as in the Sphaerulariinae (Sphaerulariidae). Tylenchids<br />
from the Neotylenchidae, Allantonematidae,<br />
lotonchiidae, and Parasitotylenchidae<br />
have insect-parasitic forms that are obese and<br />
have degenerate esophagi, intestines that are degenerate<br />
or modifed as storage organs, and the<br />
stylet often sunken into the body or even lacking<br />
(Siddiqi, 2000), suggesting that they employ<br />
some form of transcuticular or transepidermal<br />
uptake.<br />
Deladenus (Neotylenchidae), Paraiotonchium<br />
(lotonchiidae), Howardula (Allantonematidae),<br />
Skarbilovinerna (lotonchiidae), and Fergusobia<br />
(Neotylenchidae) may represent contemporary<br />
examples of an evolutionary trend from per os<br />
to transepidermal nutrient acquisition in insectparasitic<br />
Tylenchida. Of course, this is a highly<br />
speculative exercise until more information<br />
about the transition between preparasitic and<br />
parasitic females is known and some independent<br />
phylogenetic data are available. The evolutionary<br />
trend is hypothesized to be: 1) Per os<br />
acquisition via a stylet, esophagus, and gut. This<br />
strategy takes advantage of the existing stylet for<br />
feeding on fungi, plants, or other invertebrates.<br />
It is a less energy- and time-efficient method of<br />
nutrient acquisition for a hemocoelic parasite because<br />
obtaining food through the stylet requires<br />
expending energy to maintain and operate its<br />
esophagus and intestine. 2) Per os acquisition<br />
with thinning and partial apolysis of the cuticle<br />
and coincident epidermal folding to increase surface<br />
area for supplemental transcuticular uptake<br />
of nutrients (possibly Deladenus spp.). 3) Early<br />
per os acquisition followed by apolysis, partial<br />
absorption of the cuticle without the creation of<br />
a new cuticle, and folding of the epidermis such<br />
that uptake is transcuticular and somatic muscles,<br />
esophagus, and gut degenerate (e.g., Paraiotonchium).<br />
4) Early per os acquisition followed<br />
by full apolysis and ecdysis without the<br />
creation of a new cuticle. There is hypertrophy<br />
and folding of the epidermis, and nutrient uptake<br />
is transepidermal, somatic muscles and esophagus<br />
degenerate, and the gut degenerates or is<br />
transformed into a storage organ (e.g., Howardula,<br />
Skarbilovinema, and Fergusobia). The epidermal<br />
hypertrophy and folding are superficially<br />
similar to the formation of plicae (epidermal<br />
folds) during the development of a new cuticle<br />
(Bird and Bird, 1991) but are more extensive<br />
and apparently are not accompanied by the formation<br />
of a new cuticle.<br />
Acknowledgments<br />
We thank Drs. Bill Howard and Thomas<br />
Weissling for review of the manuscript and Matthew<br />
Purcell, Jeff Makinson, and Dr. John<br />
Goolsby for making the senior author's visit to<br />
the Australian Biological Control laboratory in<br />
Indooroopilly, Queensland, Australia, such a<br />
productive and enjoyable experience. This project<br />
was funded in part by USDA-ARS Specific<br />
Cooperative Agreement No. 58-6629-9-004<br />
from the USDA Invasive Plant Research Laboratory<br />
in Davie, Florida, U.S.A. This is Florida<br />
Agricultural Experiment Station Journal Series<br />
No. R-07870.<br />
Literature Cited<br />
Bird, A. F., and J. Bird. 1991. The Structure of Nematodes,<br />
2nd ed. Academic Press, Inc., New York,<br />
U.S.A. 316 pp.<br />
Chapman, R. F. 1972. The Insects: Structure and<br />
Function. American Elsevier Publishing Co., Inc.,<br />
New York, U.S.A. 819 pp.<br />
Currie, G. A. 1937. Galls on Eucalyptus trees: A new<br />
type of association between flies and nematodes.<br />
Proceedings of the Linnean Society of New South<br />
Wales 62:147-174.<br />
Giblin-Davis, R. M., K. A. Davies, G. S. Taylor, and<br />
W. K. Thomas. <strong>2001</strong>. Entomophilic nematode<br />
models for studying biodiversity and cospeciation.<br />
Pages 00-00 in Z. X. Chen, S. Y. Chen, and D.<br />
W. Dickson, eds. Nematology, Advances and Perspectives.<br />
Tsinghua University Press/Springer-<br />
Verlag, New York, U.S.A. (In press.)<br />
Maggenti, A. R. 1982. General Nematology. Springer-<br />
Verlag, New York. 372 pp.<br />
Nicholas, W. L. 1972. The fine structure of the cuticle<br />
of Heterotylenchus. Nematologica 18:138-140.<br />
Poinar, G. O., Jr. 1979. Nematodes for Biological<br />
Copyright © 2011, The Helminthological Society of Washington
248 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Control of Insects. CRC Press, Inc., Boca Raton, Subbotin, S. A., V. N. Chizhov, and N. N. Zakhar-<br />
Florida, U.S.A. 277 pp.<br />
Riding, I. L. 1970. Microvilli on the outside of a nematode.<br />
Nature 226:179-180.<br />
Siddiqi, M. R. 2000. Tylenchida: Parasites of Plants<br />
and Insects, 2nd ed. CABI Publishing, St. Albans,<br />
U.K. 848 pp.<br />
Southey, J. F., ed. 1970. Laboratory methods for work<br />
with plant and soil nematodes, 5th ed. Technical<br />
Bulletin 2 of the Ministry of Agriculture, Fisheries<br />
and Food. Her Majesty's Stationery Office, London,<br />
U.K. 148 pp.<br />
enkova. 1993. Ultrastructure of the body wall of<br />
parasitic and infective females of Skarbilovinema<br />
laumondi (Tylenchida: lotonchiidae). Fundamental<br />
and Applied Nematology 16:1-4.<br />
, , and . 1994. Ultrastructure of<br />
the integument of parasitic females in entomoparasitic<br />
tylenchids. 1. Two species of the genus<br />
Wachekitylenchus, Allantonema mirabile and Bradynema<br />
rigidum. Russian Journal of Nematology<br />
2:105-112.<br />
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Copyright © 2011, The Helminthological Society of Washington<br />
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USDA:ARS:BARC-East, No. 1180<br />
Beltsville, MD 20705-2350<br />
David J. Chitwood<br />
Nancy D. Pacheco<br />
Harley G. Sheffield
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 249-255<br />
Tegumentary Ultrastructure (SEM) of Preadult and Adult<br />
Lobatostoma jungwirthi Kritscher, 1974 (Trematoda: Aspidogastrea)<br />
ANALIA PAOLA' AND MARIA CRISTINA DAMBORENEA2<br />
Departamento Cientffico Zoologia Invertebrados, Facultad de Ciencias Naturales y Museo,<br />
Paseo del Bosque s/n° -1900- La Plata, CONICET.PIP 4728/96 Argentina<br />
(e-mail: 'apaola@museo.fcnym.unlp.edu.ar; 2cdambor@museo.fcnym.unlp.edu.ar)<br />
ABSTRACT: Larval Lobatostoma jungwirthi Kritscher, 1974 (Trematoda: Aspidogastrea) parasitize the digestive<br />
gland of Heleobia parchappii (d'Orbigny, 1835) (Mollusca: Hydrobiidae) and, as adults, the posterior intestine<br />
of the chameleon cichlid Cichlasotna facetum (Jenyns, 1842) (Pisces, Cichlidae). Currently, L. jungwirthi is the<br />
only aspidogastrid reported from freshwater fishes in Argentina. Tegumentary structures of preadults and adults<br />
of L. jungwirthi were observed under scanning electron microscopy. In the preadult, 2 types of sensory receptors<br />
were observed: monociliate papillae of intermediate length on the walls and crests of the ventral adhesive disc<br />
as well as on the disc periphery and oral lobules, and nonciliate dome-shaped papillae on the crests of the<br />
ventral adhesive disc, neck, and oral lobules. In adults, other types of sensory receptors could be observed: in<br />
the posterior dorsal region, monociliate papillae with longer cilia than those found in the preadult, and a multiciliate<br />
structure in the dorsal region at the posterior third of the body. This is the first record of a surface<br />
multiciliate receptor in aspidogastreans. The pores of marginal glands were found only between the anterior<br />
alveoli.<br />
KEY WORDS: Aspidogastrea, Lobatostoma jungwirthi, tegument, sensory papillae, SEM, Argentina.<br />
Lobatostoma jungwirthi Kritscher, 1974<br />
(Trematoda: Aspidogastrea), is the only species<br />
of the genus that parasitizes freshwater fishes. It<br />
was first found in 1974, in the stripefin eartheater<br />
Gymnogeophagus rhabdotus (Hensel,<br />
1870), in the Sinus River, Brazil (Kritscher,<br />
1974). Lunaschi (1984) found it in the posterior<br />
intestine of the chameleon cichlid Cichlasoma<br />
facetum (Jenyns, 1842) (Pisces: Cichlidae) at 2<br />
localities in Buenos Aires Province. Later, Zylber<br />
and Ostrowski de Nunez (1999) described<br />
the larval stages of L. jungwirthi from the gonad<br />
of Heleobia castellanosae (Gaillard, 1974) (Gastropoda:<br />
Hydrobiidae) collected in an artificial<br />
pond in Buenos Aires City.<br />
To date, the morphology of the larval (Zylber<br />
and Ostrowski de Nunez, 1999) and adult<br />
(Kritscher, 1974; Lunaschi, 1984) stages of this<br />
species is known only at the light microscopy<br />
level. Several investigators have described the<br />
tegumentary ultrastructure of adult aspidogastrids,<br />
such as Aspidogaster conchicola Baer,<br />
1826 (Halton and Lyness, 1971), and Cotylogaster<br />
occidentalis Nickerson, 1902 (Ip and<br />
Desser, 1984). The variability of tegumentary<br />
sensory structures of the cotylocidia of C. occidentalis<br />
(Fredericksen, 1978), the development<br />
and growth of the ventral adhesive disc of C.<br />
Corresponding author.<br />
249<br />
occidentalis and A. conchicola (Fredericksen,<br />
1980), and the sensory receptors of the larval<br />
stage of Lobatostoma manteri Rohde, 1973<br />
(Rohde and Watson, 1989a, b, 1992), and Multicotyle<br />
purvisi Dawes, 1941 (Rohde and Watson,<br />
1990b, c, d), were also studied. The aim of<br />
the present paper is to describe the tegumentary<br />
ultrastructure of juvenile and adult specimens of<br />
L. jungwirthi under scanning electron microscopy<br />
(SEM).<br />
Materials and Methods<br />
Parasites removed from the posterior intestine of C.<br />
facetum were identified as L. jungwirthi on the basis<br />
of the descriptions of Kritscher (1974) and Lunaschi<br />
(1984). Juvenile stages were found in the digestive<br />
gland of Heleobia parchappii (d'Orbigny, 1835) (Mollusca:<br />
Hydrobiidae), and adult specimens were obtained<br />
from the posterior intestine of C. facetum. Both<br />
host species were naturally parasitized by this aspidogastrid<br />
in Saladita Pond, Avellaneda District, Buenos<br />
Aires.<br />
The specimens were fixed in 10% formalin and<br />
washed in distilled water. They were dehydrated by 2<br />
changes in 35, 50, 70, and 90% acetone for 15 min<br />
each and 3 changes in 100% acetone. The material was<br />
critical point dried, then mounted on stubs and coated<br />
for SEM observation (JEOL 100).<br />
The immature stage was named the postacetabular<br />
juvenile, following the nomenclature used by Fredericksen<br />
(1980). Two stages could be distinguished according<br />
to the development of the ventral adhesive<br />
disc: a recently formed postacetabular juvenile, with<br />
little differentiation of alveoli and the buccal opening<br />
Copyright © 2011, The Helminthological Society of Washington
250 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
without oral lobules; and a preadult characterized by<br />
the presence of oral lobules and a distinct differentiation<br />
between alveoli, with the external morphology<br />
similar to that of the adult.<br />
Results<br />
Postacetabular juvenile<br />
In the recently formed postacetabular juvenile<br />
(Fig. 1), the oral disc characteristic of the adult<br />
phase is not observed. The mouth is a simple<br />
opening, without lobules (Fig. 2).<br />
The preadult shows a rough dorsal cone. Central<br />
and marginal alveoli, completely differentiated<br />
and varying in number, are observed on the<br />
ventral adhesive disc (Fig. 3). The posterior alveoli<br />
are less developed. Monociliate sensory<br />
papillae are found in the internal wall of the<br />
marginal alveoli (Fig. 4). The pores of the marginal<br />
bodies can be observed on the external<br />
border between the marginal alveoli. They are<br />
more developed in the anterior region of the<br />
ventral disc (Figs. 4, 5). The oral disc has 3 ventral<br />
and 2 dorsal lobules as in the adult, though<br />
they are not completely developed (Fig. 6).<br />
Monociliate sensory papillae and dome-shaped<br />
papillae are observed on the posterior surface of<br />
the oral disc. In this region, there is no regular<br />
distribution pattern of sensory structures (Fig.<br />
7).<br />
In the dorsal region of the neck, immediately<br />
behind the oral disc, there are pores and domeshaped<br />
papillae (Fig. 8). The monociliate papillae<br />
each have a cilium emerging from a bulbous<br />
surface.<br />
Adult<br />
The ventral adhesive disc has 16 marginal<br />
pairs and 32 central alveoli. The limit between<br />
both groups of central alveoli cannot be clearly<br />
observed (Fig. 9). The anterior region of the<br />
ventral adhesive disc shows a neat differentiation<br />
among the alveoli, with many sensory structures<br />
(Fig. 10). Completely differentiated pores<br />
of the marginal bodies are found between the<br />
marginal alveoli (Fig. 11). Monociliate papillae<br />
are located on the internal wall of each alveolus,<br />
arranged in 2 concentric circles. Two rows of<br />
dome-shaped papillae occur on the external edge<br />
of the marginal alveoli (Figs. 11-13). Monociliate<br />
and dome-shaped papillae (Fig. 14) are<br />
present on both sides of the transverse dividing<br />
line between the central alveoli. The excretory<br />
pore can be seen in the dorsal cone (Fig. 15).<br />
Copyright © 2011, The Helminthological Society of Washington<br />
Two kinds of monociliate receptors (Fig. 16) and<br />
a single multiciliate structure (Fig. 17) were observed<br />
in the posterior dorsal region.<br />
Discussion<br />
The external morphology of the preadult of L.<br />
jungwirthi is very similar to that of the adult<br />
from C. facetum. Both juveniles and adults show<br />
a single excretory pore that ends at the channel<br />
formed by the union of the lateral ducts, as described<br />
by Lunaschi (1984). In agreement with<br />
the observations of Kritscher (1974) and Lunaschi<br />
(1984), the adult stage does not show a pore<br />
of Laurer's canal.<br />
Four types of sensory receptors were observed<br />
by SEM:<br />
MONOCILIATE PAPILLAE WITH A SHORT CILIUM:<br />
This type of receptor was irregularly distributed<br />
on the dorsal tegument, in the posterior surface<br />
of the oral lobules, and in the neck of juvenile<br />
L. jungwirthi. A denned pattern of distribution<br />
was observed only on the edges of the alveoli<br />
of the ventral adhesive disc. Rohde and Watson<br />
(1992) described this structure as a receptor<br />
formed by a cilium of intermediate length, being<br />
the most common type on the surface of L. manteri.<br />
Halton and Lyness (1971) described this<br />
type of papilla as the most frequent receptor on<br />
the body surface of A. conchicola. This receptor<br />
is more abundant in the oral lobules and in the<br />
central, marginal, and peripheral regions of the<br />
ventral adhesive disc of L. jungwirthi. This distribution<br />
agrees with that observed by Halton<br />
and Lyness (1971) in A. conchicola. The type of<br />
monociliate papilla found in L. jungwirthi may<br />
also correspond to that described by Fredericksen<br />
in the juvenile acetabulum of C. occidentalis<br />
and the simple uniciliate sensory structures observed<br />
in the cotylocidium larva of the same<br />
species (Fredericksen, 1978). Likewise, they are<br />
similar to type I sensilla found in adult C. occidentalis<br />
(Ip and Desser, 1984). Monociliate receptors<br />
were observed in Lobatostoma sp. (Rohde,<br />
1972), and the type I receptor has been observed<br />
in the tegument of posterior suckerlets of<br />
larval Multicotyle purvisi (Rohde and Watson,<br />
1990c). Monociliate tegumental receptors were<br />
also found in the buccal complex of Polylabroides<br />
australis (Murray, 1931) (Monogenea, Microcotylidae)<br />
(Rohde and Watson, 1995b) and in<br />
Udonella sp. (Platyhelminthes) (Rohde et al.,<br />
1989).<br />
MONOCILIATE PAPILLAE WITH A LONG CILIUM:
PAOLA AND DAMBORENEA—TEGUMENT OF LOBATOSTOMA JUNGWIRTHI 251<br />
Figures 1-5. Immature stages of Lobatostoma jungwirthi. 1. Postacetabular juvenile showing a simple<br />
buccal cavity (without lobules) and the ventral adhesive disc with poorly differentiated alveoli. 2. Frontal<br />
view of the buccal cavity of the postacetabular juvenile. 3. Alveoli of the ventral adhesive disc, oral lobules,<br />
and a rough posterior cone (dorsal) (black arrow) in latero-ventral view of the preadult. 4. Marginal<br />
alveoli of the ventral adhesive disc of the preadult, with sensory receptors of the monociliate type (white<br />
arrow), dome-shaped receptor (arrowhead), and the pores of the marginal glands not yet formed between<br />
the posterior alveoli (black arrow). 5. Pore of a marginal gland (^marginal body) of the preadult. Scales:<br />
1, 2 = 10 |xm; 3 = 50 fim; 4 = 25 |xm; 5 = 10 |xm.<br />
These were found only on the surface of the<br />
dorsal cone of the adult. This type of receptor<br />
may correspond to the receptor with a long cilium<br />
in the larva of L. manteri (Rohde and Watson,<br />
1992). It is also similar to the receptor type<br />
A described by Ip et al. (1982) in adult C. occidentalis.<br />
NONCILIATE DOME-SHAPED RECEPTORS: These<br />
were found on the posterior surface of the oral<br />
lobules and in the neck of the juvenile. They<br />
Copyright © 2011, The Helminthological Society of Washington
252 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Figures 6-10. Preadult and adult of Lobatostoma jungwirthi. 6. Frontal view of the preadult showing<br />
the 5 oral lobules. 7. Posterior surface of the oral lobules showing monociliate papillae (arrow) and domeshaped<br />
papillae arranged without defined pattern. 8. Neck region (ventral) with pores (white arrow) (in<br />
some cases with secretion) and dome-shaped papillae. 9. General ventral view of the adult. A clear differentiation<br />
of the longitudinal septum of the ventral adhesive disc cannot be observed. 10. Anterior end<br />
of the ventral adhesive disc showing dome-shaped and monociliate papillae (arrow) on the edge of the<br />
walls. Scales: 6 = 50 u,m; 7, 8 = 10 u,m; 9 = 25 u,m; 10 = 100 u,m.<br />
also were distributed on the borders between the<br />
alveoli of the adhesive disc of both juvenile and<br />
adult worms. Nonciliate sensory receptors were<br />
found by Rohde and Watson (1990b) in the external<br />
ventral tegument of the ventral suckerlets<br />
in M. purvisi. This structure may correspond to<br />
the nonciliate disc-shaped receptors or to the<br />
Copyright © 2011, The Helminthological Society of Washington<br />
nonciliate type with a long root described by<br />
Rohde and Watson (1992) in the larva of L.<br />
manteri. Nonciliate tegumental receptors were<br />
also found in the juvenile of Astramphilina elongata<br />
Johnston, 1931 (Monogenea) (Rohde and<br />
Watson, 1990a).<br />
A SINGLE MULTICILIATE RECEPTOR: This Was
PAOLA AND DAMBORENEA—TEGUMENT OF LOBATOSTOMA JUNGWIRTHI 253<br />
Figures 11-14. Ventral adhesive disc of adult Lobatostoma jungwirthi. 11. General view of the marginal<br />
alveoli on the marginal region at midbody level of the ventral adhesive disc. 12. Walls of the marginal<br />
alveoli showing 2 circles of dome-shaped papillae (arrow) and marginal gland pore. 13. Detail of the<br />
internal wall with almost 1 complete circle of monociliate papillae (arrows). 14. Ventral view: transverse<br />
septum with monociliate (black arrow) and nonciliate papillae (white arrow). Scales: 11 = 50 u,m; 12 =<br />
25 u,m; 13, 14 = 10 jrni.<br />
observed in the posterior third of the dorsal region<br />
of the adult of L. jungwirthi. Paired multiciliate<br />
receptor complexes with 10 short cilia<br />
were described by Rohde and Watson (1990d)<br />
to be located dorsally to the mouth cavity of<br />
larval M. purvisi, but our record is the first of a<br />
surface multiciliate receptor in aspidogastreans.<br />
However, multiciliate receptors are found in other<br />
groups of parasitic Platyhelminthes, i.e., in the<br />
taste organ of the buccal complex of Pricea tnultae<br />
Chauhan, 1945 (Monogenea, Gastrocotylidae).<br />
Rohde and Watson (1996) found these receptors<br />
concentrated in small pits. Additionally,<br />
multiciliate pit-receptors were observed in the<br />
buccal complex of Polylabroides australis (Rohde<br />
and Watson, 1995b) and in specimens of an<br />
undescribed species of Proseriata (Monocelididae:<br />
Minonidae) (Rohde and Watson, 1995a).<br />
Pores of the marginal bodies were observed<br />
on the external border between the marginal alveoli<br />
of juveniles and adults of L. jungwirthi.<br />
Kritscher (1974) found similar structures in the<br />
adult of the species. These pores were described<br />
as part of a glandular system in the adhesive disc<br />
of aspidogastrids (Rohde, 1994).<br />
Unlike the preadult, the youngest juvenile<br />
does not have oral lobules. A few less-developed<br />
alveoli were observed on the ventral adhesive<br />
disc. Two sensory receptors were found in the<br />
preadult of L. jungwirthi'. a ciliate receptor with<br />
a cilium of intermediate length, and a nonciliate<br />
dome-shaped receptor. These correspond to 2 of<br />
Copyright © 2011, The Helminthological Society of Washington
254 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Figures 15-17. Tegument of posterior dorsal region<br />
of Lobatostoma jungwirthi. 15. Excretory pore<br />
on the dorsal cone. 16. Type of monociliate papilla<br />
in the posterior cone region. 17. Multiciliate sensory<br />
receptor in the posterior dorsal region. Scales: 15,<br />
17 = 5 (Jim; 16 = 10 fjim.<br />
the 4 types of receptors described by Rohde and<br />
Watson (1992) in the larva of L. manteri. In addition,<br />
a third kind of receptor was observed that<br />
corresponds to the marginal body complex in the<br />
adult. However, the complex formed by the marginal<br />
bodies was observed as described for the<br />
adult, though the pores of these glands are not<br />
yet developed in the posterior region of the ventral<br />
adhesive disc in the preadult.<br />
The most common receptors in the preadult<br />
Copyright © 2011, The Helminthological Society of Washington<br />
and in the adult were the intermediate length<br />
monociliate and the nonciliate dome-shaped papillae.<br />
The monociliate structures may correspond<br />
to some of the types I to V of monociliate<br />
dome-shaped papillae described for adult L.<br />
manteri (Rohde and Watson, 1989a). The nonciliate<br />
dome-shaped papillae may be homologous<br />
to those of type VI A and B found in the<br />
adult of L. manteri. Both types of receptors were<br />
abundant over the entire body, arranged in circles<br />
on the ventral adhesive disc. Longer monociliate<br />
papillae and multiciliate receptors were<br />
found exclusively in the adult. Probably more<br />
than one type of nonciliate and monociliate papillae<br />
may exist. It would be interesting to study<br />
these papillae further under transmission electron<br />
microscopy.<br />
Literature Cited<br />
Fredericksen, D. 1978. The fine structure and phylogenetic<br />
position of the cotylocidium larva of Cotylogaster<br />
occidentalis Nickerson, 1902 (Trematoda,<br />
Aspidogastrea). Journal of <strong>Parasitology</strong> 64:<br />
961-966.<br />
. 1980. Development of Cotylogaster occidentalis<br />
Nickerson, 1902 (Trematoda, Aspidogastridae)<br />
with observations on the growth of the ventral<br />
adhesive disc in Aspidogaster conchicola<br />
Baer, 1827. Journal of <strong>Parasitology</strong> 66:973-984.<br />
Halton, D. W., and R. A. W. Lyness. 1971. Ultrastructure<br />
of the tegument and associated structures<br />
of Aspidogaster conchicola (Trematoda: Aspidogastrea).<br />
Journal of <strong>Parasitology</strong> 57:1198-1210.<br />
Ip, H., and S. Desser. 1984. Transmission electron<br />
microscopy of the tegumentary sense organs of<br />
Cotylogaster occidentalis (Trematoda: Aspidogastrea).<br />
Journal of <strong>Parasitology</strong> 70:563-575.<br />
, , and I. Weller. 1982. Cotylogaster<br />
occidentalis (Trematoda, Aspidogastrea): scanning<br />
electron microscopic observations of sense organs<br />
and associated surface structures. Transactions of<br />
the American Microscopical Society 101:253-<br />
261.<br />
Kritscher, V. E. 1974. Lobatostoma jungwirthi nov.<br />
spec. (Aspidocotylea, Aspidogastridae) aus Geophagus<br />
brachiurus Cope 1894 (Pise., Cichlidae).<br />
Annalen des Naturhistorischen Museums in Wien<br />
78:381-384.<br />
Lunaschi, L. 1984. Helmintos parasites de peces de<br />
agua dulce de Argentina. II. Presencia de Lobatostoma<br />
jungwirthi Kritscher, 1974 (Trematoda:<br />
Aspidogastrea) en Cichlasoma facetum (Jenyns).<br />
Neotropica 20:187-193.<br />
Rohde, K. 1972. Sinnesrezeptoren von Lobatostoma<br />
n. sp. (Trematoda, Aspidogastrea). Die Naturwissenschaften<br />
59:1<strong>68</strong>-169.<br />
. 1994. The Aspidogastrea, especially Multicotyle<br />
purvisi Dawes, 1941. Advances in <strong>Parasitology</strong><br />
10:78-151.<br />
, and N. Watson. 1989a. Sense receptors in
PAOLA AND DAMBORENEA—TEGUMENT OF LOBATOSTOMA JUNGWIRTH1 255<br />
Lobatostoma manteri (Trematoda, Aspidogastrea).<br />
International Journal for <strong>Parasitology</strong> 19:847-858.<br />
, and . 1989b. Ultrastructure of the marginal<br />
glands of Lobatostoma manteri (Trematoda,<br />
Aspidogastrea). Zoologischer Anzeiger 223:301-<br />
310.<br />
, and . 1990a. Ultrastructural studies of<br />
juvenile Austramphilina elongata: transmission<br />
electron microscopy of sensory receptors. <strong>Parasitology</strong><br />
Research 76:336-342.<br />
, and . 1990b. Non-ciliate sensory receptors<br />
of larval Multicotyle purvisi (Trematoda,<br />
Aspidogastrea). <strong>Parasitology</strong> Research 76:585-<br />
590.<br />
, and . 1990c. Uniciliate sensory receptors<br />
of larval Multicotyle purvisi (Trematoda, Aspidogastrea).<br />
<strong>Parasitology</strong> Research 76:591-596.<br />
, and . 1990d. Paired multiciliate receptor<br />
complexes in larval Multicotyle purvisi (Trematoda,<br />
Aspidogastrea). <strong>Parasitology</strong> Research 76:<br />
597-601.<br />
, and . 1992. Sense receptors of larval<br />
Lobatostoma manteri (Trematoda, Aspidogastrea).<br />
International Journal for <strong>Parasitology</strong> 22:35-42.<br />
New Books Available<br />
-, and -. 1995a. Sensory receptors and<br />
epidermal structures of a meiofaunal turbellarian<br />
(Proseriata: Monocelididae: Minoninae). Australian<br />
Journal of Zoology 43:69-81.<br />
, and . 1995b. Ultrastructure of the buccal<br />
complex of Polylabroides australis (Monogenea,<br />
Polyopisthocotylea, Microcotylidae). International<br />
Journal for <strong>Parasitology</strong> 25:307-318.<br />
, and . 1996. Ultrastructure of the buccal<br />
complex of Pricea multae (Monogenea, Polyopisthocotylea,<br />
Gastrocotylidae). Folia Parasitologica43:117-132.<br />
-, and F. Roubal. 1989. Ultrastructure<br />
of flame bulbs, sense receptors, tegument and<br />
sperm of Udonella (Platyhehninthes) and the phylogenetic<br />
position of the genus. Zoologischer Anzeiger<br />
222:143-157.<br />
Zylber, M. I., and M. Ostrowski de Nunez. 1999.<br />
Some aspects of the development of Lobatostoma<br />
jungwirthi Kritscher, 1974 (Aspidogastrea) in<br />
snails and cichlids fishes from Buenos Aires, Argentina.<br />
Memorias do Instituto Oswaldo Cruz 94:<br />
31-35.<br />
The Flagellates: Unity, Diversity and Evolution. Barry S. C. Leadbeater and J. C. Green, Editors. 2000.<br />
The Systematics Association Special Volume Series 59, Taylor and Francis Limited, 29 West 35th Street,<br />
New York, NY 10001-2299. i-xi, 401 pp. ISBN 0-7484-0914-9. 7" X 9%", hard cover. Cost: US$130.00<br />
or Canadian $195.00, per copy plus shipping and handling. Abstract: 35 Authors have contributed to the<br />
17 chapters of this examination of the blood parasite group. "[The] book sets out to examine flagellates<br />
from a multidisciplinary standpoint. Of primary concern are the unifying structures, mechanisms and<br />
processes involved in flagellate biology. [It begins] with a review of the complex history of flagellate<br />
studies from the first use of microscopes ... to the present. [This is] followed by a series of chapters on<br />
common aspects of flagellates, . . . [including] a discussion of the problems inherent in being a flagellate,<br />
and reviews of the structure and function of the flagellum itself, the cytoskeleton, surface structures and<br />
sensory mechanisms. The diversity of flagellates is recognized in the next series of chapters, which include<br />
reviews of trophic strategies of both free-living and parasitic groups, and contributions on ecology, biogeography<br />
and population genetics. The final chapters . . . are concerned with the occurrence and loss of<br />
organelles, and other aspects of flagellate evolution and phylogeny."<br />
Interrelationships of the Platyhelminths. D. T. J. Littlewood and R. A. Bray, Editors. <strong>2001</strong>. The Systematics<br />
Association Special Volume Series 60, Taylor and Francis Limited, 29 West 35th Street, New<br />
York, NY 10001-2299. i-xii, 356 pp. ISBN 0-7484-0903-3. 8W X 11", hard cover. Cost: US$125.00 or<br />
Canadian $188.00, per copy plus shipping and handling. Abstract: This book's 27 chapters have been<br />
prepared by no fewer than 50 different expert contributors. It "has been split into four sections, rather<br />
dissimilar in length, that highlight the underlying goals [of bringing together workers from divergent areas<br />
to produce a work unified by modern approaches to phylogenetic analysis]. The first section takes a broader<br />
perspective on the status of the Platyhelminthes, its monophyly, placement in relation to other Metazoa<br />
and the nature of the basal taxa. The second section deals with the interrelationships of major free-living<br />
taxa, and the third on symbiotic and parasitic taxa. The final section encompasses contributions that view<br />
phylogeny and phylogenetic inference from the point of view of particular characters or techniques."<br />
Copyright © 2011, The Helminthological Society of Washington
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 256-259<br />
Research Note<br />
Infectivity and <strong>Comparative</strong> Pathology of Echinostoma caproni,<br />
Echinostoma revolutum, and Echinostoma trivolvis (Trematoda) in the<br />
Domestic Chick<br />
SHANNON K. MULLIGAN,' JANE E. HUFFMAN,1-3 AND BERNARD FRIED2<br />
1 Department of Biological Sciences, Fish and Wildlife Microbiology Laboratory, East Stroudsburg University,<br />
East Stroudsburg, Pennsylvania 18301, U.S.A. (e-mail: jhuffman@po-box.esu.edu) and<br />
2 Department of Biology, Lafayette <strong>College</strong>, Easton, Pennsylvania 18042, U.S.A.<br />
ABSTRACT: We examined the clinical and pathological<br />
effects of 3 species of 37-collar-spined Echinostoma in<br />
domestic chicks. Three groups of 6 chicks each were<br />
infected with 50 metacercariae of either Echinostoma<br />
caproni, Echinostoma revolutum, or Echinostoma trivolvis.<br />
A group of 6 chicks was not infected and<br />
served as the uninfected controls. The chicks were<br />
necropsied on day 14 postinfection (PI). Infectivity and<br />
worm recovery rates for E. caproni were 100% and<br />
24%, respectively; for E. revolutum, they were 67%<br />
and 9%, respectively; and for E. trivolvis, they were<br />
83% and 15%, respectively. Echinostoma caproni was<br />
located in the middle third of the small intestine,<br />
whereas E. revolutum and E. trivolvis were located in<br />
the lower third, showing that niche selection of the<br />
different echinostomes varied. The echinostomes became<br />
ovigerous on days 10, 12, and 14 PI for E. caproni,<br />
E. trivolvis, and E. revolutum, respectively. Goblet<br />
cell proliferation in the host intestinal mucosa occurred<br />
in all infections.<br />
KEY WORDS: Echinostoma caproni, Echinostoma<br />
trivolvis, Echinostoma revolutum, Trematoda, domestic<br />
chicks, echinostomiasis, pathology, clinical effects,<br />
goblet cell, infectivity.<br />
Because echinostomiasis has produced significant<br />
mortality in ducks raised for commercial<br />
production in Europe and Asia (Kishore and<br />
Sinha, 1982), studies on experimental avian<br />
models to define the clinical and pathological<br />
features of the echinostomes are needed. Except<br />
for the experimental studies by Kim and Fried<br />
(1989) on gross and histopathological effects of<br />
Echinostoma caproni Richard, 1964, in an experimental<br />
avian model, such studies are lacking.<br />
In North America, avian hosts in the wild are<br />
often infected with Echinostoma trivolvis (Cort,<br />
1914) and Echinostoma revolutum (Froelich,<br />
1802) and species of Echinoparyphium (43- and<br />
3 Corresponding author.<br />
256<br />
Copyright © 2011, The Helminthological Society of Washington<br />
45-collar-spined echinostomes). Interestingly,<br />
the habitat of species of Echinoparyphium in the<br />
gut of birds is more anteriad than that of either<br />
E. trivolvis or E. revolutum. Echinostoma caproni<br />
also tends to localize more anteriad in the<br />
avian gut than either E. trivolvis or E. revolutum,<br />
and may serve as a useful model for Echinoparyphium<br />
infections. Therefore, information<br />
obtained from single infections of the 3 echinostome<br />
species examined in this study may be<br />
useful to wildlife studies of birds naturally infected<br />
with 3 or more species of echinostomes.<br />
The objectives of this study were to determine<br />
the following parameters in E. caproni', E. revolutum-,<br />
and E. trivolvis-infected birds: packed<br />
cell volume, hemoglobin concentration, and the<br />
relative splenic and hepatic weights of infected<br />
and noninfected domestic chicks. Parasite recovery<br />
and location were recorded from infected<br />
animals. We also examined tissues grossly and<br />
microscopically for evidence of pathological<br />
changes. Metacercarial cysts of E. caproni and<br />
E. trivolvis were obtained from the kidneys and<br />
pericardial sacs of laboratory-infected Biornphalaria<br />
glabrata (Say, 1816) snails (Huffman<br />
and Fried, 1990). Metacercarial cysts of E. revolutum<br />
were obtained from experimentally infected<br />
Lymnaea elodes (Say, 1821) snails (Sorenson<br />
et al., 1997). Twenty-four-d-old unfed<br />
domestic chicks were obtained from Reich Poultry<br />
Farm (Marietta, Pennsylvania, U.S.A.). All<br />
chicks were infected on day 1 prior to feeding.<br />
All animals were provided food (Country Egg<br />
Producer®, Agway Inc., Syracuse, New York,<br />
U.S.A.) and water ad libitum throughout the<br />
study. Group A (N = 6) was not infected and<br />
served as controls for the study. Chicks in<br />
Groups B-D each received 50 metacercarial<br />
cysts per os of either E. caproni (Group B, N =<br />
6), E. trivolvis (Group C, N = 6), or E. revolu-
RESEARCH NOTES 257<br />
Table 1. Mean worm recovery, mean percentage of recovery, percentage infected, and range of parasites<br />
recovered from chicks infected with Echinostoma caproni (Group B, TV = 6), Echinostoma trivolvis (Group<br />
C, N = 6), and Echinostoma revolutum (Group D, N = 6). Uninfected controls are represented by Group<br />
A (TV = 6).<br />
Group<br />
A<br />
B<br />
C<br />
D<br />
No. cysts<br />
administered<br />
0<br />
50<br />
50<br />
50<br />
Mean no. worms<br />
recovered (range)<br />
0(0)<br />
12 (8-20)<br />
8 (0-21)<br />
5 (0-16)<br />
turn (Group D, /V = 6). Fecal samples were collected<br />
from each infected animal and checked<br />
for echinostome eggs daily, starting on day 8<br />
postinfection (PI). Approximately 1 g of feces<br />
was emulsified in distilled water and examined<br />
by light microscopy. Animals were weighed every<br />
2 d to monitor weight gain in the chicks.<br />
Blood samples were collected from the jugular<br />
veins of all chicks on day 14 PI into tubes<br />
containing 0.13 M sodium citrate, refrigerated,<br />
and processed within 24 hr. Packed cell volume<br />
and hemoglobin concentration were measured<br />
and recorded.<br />
All chicks (Groups A-D) were necropsied on<br />
day 14 PI. The small intestines, ceca, and cloacas<br />
were opened, and the location and number<br />
of echinostomes in the infected chicks were recorded.<br />
Hepatic, splenic, and intestinal tissue<br />
samples from all groups were fixed in 10% neutral<br />
buffered formalin for 48 hr and then dehydrated<br />
in a series of graded alcohols, cleared in<br />
xylene, embedded in paraffin, and sectioned at<br />
6 (xm. At necropsy, the relative spleen and liver<br />
weights were determined.<br />
Infected and control tissues were stained in<br />
hematoxylin and eosin to evaluate histopathological<br />
effects. The occurrence of immunological<br />
cells from the infected and control tissues<br />
was also evaluated.<br />
Echinostome eggs were first seen in the fecal<br />
samples of all chicks from Group B on day 10,<br />
of 5 chicks from Group C on day 12, and of 4<br />
chicks from Group D on day 14 PI. All chicks<br />
exposed to E. caproni became infected. The<br />
number of worms recovered from their intestines<br />
ranged from 8 to 20, with a 24% recovery. The<br />
number of parasites recovered from Group C<br />
ranged from 0 to 21, a 15% recovery. Four of 6<br />
chicks (67%) exposed to E. revolutum cysts<br />
were infected, and the numbers of worms recov-<br />
Mean percentage<br />
of worms recovered<br />
0<br />
24<br />
15<br />
9<br />
Day of first<br />
appearance<br />
of eggs in feces<br />
0<br />
10<br />
12<br />
14<br />
Percentage of<br />
chicks infected<br />
0<br />
100<br />
83<br />
67<br />
ered ranged from 0 to 16, averaging 9%. These<br />
data are summarized in Table 1.<br />
The locations of the recovered worms from<br />
the infected chicks varied according to species.<br />
Echinostoma caproni was located in the midthird<br />
of the intestine, between the pylorus and<br />
the cloaca. Both E. trivolvis and E. revolutum<br />
were found toward the end of the intestine near<br />
the cloaca, but typically, E. revolutum was found<br />
more posteriad than E. trivolvis. No differences<br />
were noted between the number of parasites recovered<br />
per chick and the location of the worms.<br />
Echinostoma caproni tended to be in groups, but<br />
single worms were also found. Echinostoma revolutum<br />
and E. trivolvis were found singly and<br />
in groups.<br />
There was no significant weight loss (P ><br />
0.05) in chicks infected with any species of echinostome<br />
versus the uninfected (control) group.<br />
No differences were seen in liver or spleen<br />
weights of the infected chicks versus the control<br />
group, nor were there differences in liver or<br />
spleen weights between the infected groups.<br />
There were no notable differences in either the<br />
measured packed cell volume or hemoglobin<br />
concentration of the infected chicks compared<br />
with the uninfected chicks.<br />
Histologically, the liver and spleen tissues of<br />
Groups B-D showed no sign of immunological<br />
response or damage from the parasites. Damage<br />
to the intestinal villi of chicks infected with E.<br />
caproni was observed at the site of parasite attachment,<br />
villi were atrophic, and the circular<br />
musculature was hypertrophied with collagenlike<br />
fibers present. There was a proliferation of<br />
goblet cells. Hemorrhage occurred at the site of<br />
attachment. In chicks infected with E. trivolvis,<br />
damage to villi also occurred, with lymphocytic<br />
infiltration and goblet cell proliferation but with<br />
no hemorrhage noted. The response to E. revo-<br />
Copyright © 2011, The Helminthological Society of Washington
258 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
lutuin was less severe than with the other 2 parasites,<br />
but damage to the intestinal villi was observed.<br />
The presence of eggs in the host's feces is<br />
used for diagnosis of echinostomiasis. The time<br />
of deposition of eggs in the feces will vary<br />
among species (Huffman and Fried, 1990). In<br />
this study, E. caproni eggs were first noted in<br />
the chick's feces on day 10 PI, followed by E.<br />
trivolvis eggs on day 12 PI, and eggs of E. revolutum<br />
were found on day 14 PI.<br />
Infectivity in the chicks varied between the<br />
different Echinostoma species in this study. Factors<br />
such as age, size of cyst inoculum, pretreatment<br />
of metacercarial cysts, and host-gut emptying<br />
time influence the infectivity of E. trivolvis<br />
in experimentally infected chicks (Fried et al.,<br />
1997). Huffman and Fried (1990) reported E.<br />
trivolvis to have infectivity varying between 50<br />
and 69%. Fried (1984) reported 100% infectivity<br />
when preselected cysts were used. In this study,<br />
the E. trivolvis metacercariae administered to the<br />
chicks averaged 83% infectivity. Experimental<br />
infection with E. caproni cysts in this study resulted<br />
in 100% infectivity, agreeing with a previous<br />
study on this species conducted by Fried<br />
et al. (1988), which reported 97% infectivity.<br />
Echinostoma revolutum infectivity in this study<br />
(67%) is congruent with the report of Humphries<br />
et al. (1997) on the species in experimentally<br />
infected chicks (64%).<br />
Huffman and Fried (1990) summarized findings<br />
of average worm recoveries for E. trivolvis<br />
in experimentally infected chicks. These results<br />
varied between 6 and 21%. In this study, a mean<br />
of 15% of the flukes were recovered from chicks<br />
infected with E. trivolvis. Echinostoma caproni<br />
infections resulted in 24% worm recovery, concurring<br />
with the report by Fried et al. (1988) of<br />
28% worm recovery. An average of 9% of the<br />
administered cysts were recovered as adult<br />
worms in the E. revolutum infections. This differed<br />
from the 32% worm recovery of E. revolutum<br />
reported by Humphries et al. (1997) and<br />
the 21 % worm recovery reported for the same<br />
species of Echinostoma in domestic chicks<br />
(Fried et al., 1997).<br />
Echinostoma trivolvis distribution along the<br />
intestine of the domestic chick varied in numerous<br />
past studies (Huffman and Fried, 1990). On<br />
day 14 PI of our study, E. trivolvis was found<br />
mainly in the lower intestine near the cloaca.<br />
Echinostoma revolutum also was seen in the<br />
Copyright © 2011, The Helminthological Society of Washington<br />
posterior aspect of the intestine, congruent with<br />
results by Humphries et al. (1997) and Fried et<br />
al. (1997). Echinostoma caproni was found<br />
more anteriad than the other echinostomes in<br />
this study, mainly clustered in the midthird of<br />
the intestine.<br />
Weight gain, spleen and liver weights, packed<br />
cell volume, and hemoglobin concentrations of<br />
the infected chicks compared with the controls<br />
were not affected by the presence of any of the<br />
echinostomes. As noted in previous studies, liver<br />
or spleen tissue damage did not occur. Huffman,<br />
Iglesias, and Fried (1986) noted increased pathology<br />
in golden hamsters infected with echinostomes<br />
and increased pathology when greater<br />
numbers of parasites infected the host. Infectivity<br />
in the present study was less compared with<br />
other studies. In the study by Fried and Wilson<br />
(1981), high worm burdens caused a decrease in<br />
chick weight. Changes in blood parameters and<br />
tissue damage (Huffman, Michos, and Fried,<br />
1986) have been noted in rodent hosts infected<br />
with echinostomes.<br />
In conclusion, there are differences in the<br />
host—parasite relationships for each of the echinostomes<br />
used in this study. An understanding<br />
of these differences will contribute to better understanding<br />
the biosystematics of the 37-collarspined<br />
echinostome group. Some of these differences<br />
may help elucidate species distinctions<br />
when echinostomes are recovered from naturally<br />
infected hosts in both single and multiple infections.<br />
Literature Cited<br />
Fried, B. 1984. Infectivity, growth, and development<br />
of Echinostoma revolutum (Trematoda) in the domestic<br />
chick. Journal of Helminthology 58:241-<br />
244.<br />
, R. A. Donovick, and S. Emili. 1988. Infectivity,<br />
growth, and development of Echinostoma<br />
liei (Trematoda) in the domestic chick. International<br />
Journal for <strong>Parasitology</strong> 18:413-414.<br />
, T. J. Mueller, and B. A. Frazer. 1997. Observations<br />
of Echinostoma revolutum and Echinostoma<br />
trivolvis in single and concurrent infections<br />
in domestic chicks. International Journal for<br />
<strong>Parasitology</strong> 27:1319-1322.<br />
-, and B. D. Wilson. 1981. Decrease in body<br />
weight of domestic chicks infected with Echinostoma<br />
revolutum (Trematoda) or Zygocotyle lunata<br />
(Trematoda). Proceedings of the Helminthological<br />
Society of Washington 48:97-98.<br />
Huffman, J. E., and B. Fried. 1990. Echinostoma and<br />
echinostomiasis. Advances in <strong>Parasitology</strong> 29:<br />
215-269.
, D. Iglesias, and B. Fried. 1986. Echinostoma<br />
revolutum pathology of intestinal infections of the<br />
golden hamster. International Journal for <strong>Parasitology</strong><br />
18:873-874.<br />
-, C. Michos, and B. Fried. 1986. Clinical and<br />
pathological effects of Echinostoma trivolvis (Digenea:<br />
Echinostomatidae) in the golden hamster,<br />
Mesocricetus auratus. <strong>Parasitology</strong> 93:505=515.<br />
Humphries, J. E., A. Reddy, and B. Fried. 1997.<br />
Infectivity and growth of Echinostoma revolutum<br />
(Froelich, 1802) in the domestic chick. International<br />
Journal for <strong>Parasitology</strong> 27:129-130.<br />
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 259-261<br />
Research Note<br />
RESEARCH NOTES 259<br />
Kim, J., and B. Fried. 1989. Pathological effects of<br />
Echinostoma caproni (Trematoda) in the domestic<br />
chick. Journal of Helminthology 63:227-230.<br />
Kishore, N., and D. P. Sinha. 1982. Observations of<br />
Echinostoma revolutum infection in the rectum of<br />
domestic ducks (Anas platyrhynchos domesticiis).<br />
Agricultural Science Digest 2:57-60.<br />
Sorenson, R. E., I. Kanev, B. Fried, and D. Minchella.<br />
1997. The occurrence and identification of<br />
Echinostoma revolutum from North American<br />
Lymnaea elodes snails. Journal of <strong>Parasitology</strong> 83:<br />
169-170.<br />
Excystation and Distribution of Metacercariae of Echinostoma<br />
caproni in ICR Mice<br />
BERNARD FRIED,' ADITYA REDDY, AND CAROLINE D. BALFOUR<br />
Department of Biology, Lafayette <strong>College</strong>, Easton, Pennsylvania 18042, U.S.A. (e-mail: friedb@lafayette.edu)<br />
ABSTRACT: In vivo excystation and distribution of<br />
newly excysted metacercariae of Echinostoma caproni<br />
Richard, 1964, were studied in 16 ICR mice, each fed<br />
400 metacercarial cysts and necropsied at various intervals<br />
from 1 to 24 hr postinfection (p.i.). Excysted<br />
metacercariae were recovered from the stomach and<br />
intestine (duodenum and jejunum) at 1 hr p.i. In vivo<br />
excystation in this echinostome occurred in the stomach<br />
and the anterior part of the small intestine. Encysted<br />
metacercariae were recovered from the stomach,<br />
small intestine, and cecum-large intestine at 1 and<br />
2 hr p.i. Recovery of encysted metacercariae was rare<br />
at 3 hr and nil at 4 and 24 hr. At 3, 4, and 24 hr, the<br />
encysted metacercariae had either excysted or were<br />
voided. Excysted metacercariae were widely scattered<br />
throughout the small intestine at all times, with about<br />
75% located in segments 1, 2, and 3 (duodenal-jejunum<br />
zone) of the small intestine at 3, 4, and 24 hr p.i.<br />
KEY WORDS: trematodes, in vivo excystation, Echinostoma<br />
caproni, metacercariae, ICR mice.<br />
Although information is available (Fried and<br />
Emili, 1988; Fried, 1994; Ursone and Fried,<br />
1995) on chemical excystation of metacercarial<br />
cysts of Echinostoma caproni Richard, 1964,<br />
there are no studies on in vivo excystation of<br />
this echinostome in mice. Metacercariae of most<br />
intestinal digeneans excyst in the vertebrate<br />
Corresponding author.<br />
small intestine, but details on in vivo excystation<br />
and the microhabitat where excystation occurs<br />
are poorly understood in the Digenea. Simonsen<br />
et al. (1989) stated that E. caproni metacercarial<br />
cysts excysted in the duodenum of the mouse<br />
and the newly excysted metacercariae migrated<br />
to the posterior third of the small intestine. However,<br />
Simonsen et al. did not do experimental<br />
studies on in vivo excystation of the metacercarial<br />
cyst.<br />
The purpose of this research was to examine<br />
in vivo excystation of E. caproni metacercariae<br />
at various times up to 24 hr postinfection (p.i.)<br />
and to determine the distribution of newly excysted<br />
metacercariae in ICR mice. The ICR<br />
mouse is widely used as a laboratory host for<br />
this echinostome (see review in Fried and Huffman,<br />
1996). The only previous study that has<br />
reported distribution of preovigerous worms of<br />
E. caproni is that of Manger and Fried (1993),<br />
who showed that, by 2 days p.i., more than 90%<br />
of the juvenile worms were located in segment<br />
3 posterior to the pylorus (equivalent to the jejunum)<br />
in the ICR mouse.<br />
Metacercarial cysts of E. caproni were removed<br />
from the pericardial cavity and kidney of<br />
experimentally infected Biomphalaria glabrata<br />
Say, 1818, snails and fed (400 cysts/mouse) via<br />
stomach tube to 16 6-8-wk old, outbred, female<br />
Copyright © 2011, The Helminthological Society of Washington
260 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Table 1. Percentage of encysted (EN) and excysted (EX) metacercariae (M) of Echinostoma caproni from<br />
16 mice, each fed 400 cysts.<br />
Time<br />
postinfection<br />
Group* (hr)<br />
A<br />
B<br />
C<br />
D<br />
E<br />
1<br />
2<br />
3<br />
4<br />
24<br />
M<br />
EN<br />
EX<br />
EN<br />
EX<br />
EN<br />
EX<br />
EN<br />
EX<br />
EN<br />
EX<br />
Stomach<br />
0.9<br />
1.0<br />
0.3<br />
0.8<br />
0<br />
0.5<br />
0<br />
0.2<br />
0<br />
0<br />
1<br />
1.5<br />
1.9<br />
0.5<br />
2.4<br />
0<br />
2.6<br />
0<br />
2.3<br />
0<br />
1.6<br />
Segments of the small intestine<br />
2<br />
1.3<br />
1.6<br />
0.5<br />
1.8<br />
0.1<br />
2.9<br />
0<br />
2.6<br />
0<br />
2.7<br />
3<br />
0.7<br />
2.5<br />
0.4<br />
1.3<br />
0<br />
3.1<br />
0<br />
2.9<br />
0<br />
3.3<br />
* Groups A and B each with 2 mice; Groups C, D, and E each with 4 mice.<br />
ICR mice (Hosier and Fried, 1991). Preliminary<br />
studies on 2 mice each fed 100 metacercarial<br />
cysts and necropsied at 1 and 2 hr p.i. showed<br />
metacercarial recoveries (combined data of excysted<br />
and encysted metacercariae) of about<br />
10% at necropsy. The preliminary work showed<br />
the inherent difficulties in recovering these small<br />
organisms (excysted metacercariae measuring<br />
about 250 (xm in length and encysted metacercariae<br />
about 150 (Jim in diameter) from the intestinal<br />
tract. Moreover, the color, size, and motility<br />
of the villi made it difficult to distinguish<br />
them from excysted metacercariae. Empty cysts<br />
were also seen at necropsy but were not counted.<br />
On the basis of our experiences with the preliminary<br />
study, we increased the cyst inoculum to<br />
400 per host in the study reported herein.<br />
Groups of 2 mice each were necropsied at 1<br />
and 2 hr p.i. (Groups A and B, respectively; Table<br />
1), and groups of 4 mice each were necropsied<br />
at 3, 4, and 24 hr p.i. (Groups C, D, and E,<br />
respectively; Table 1). The numbers of encysted<br />
and excysted metacercariae in the stomach, in 5<br />
intestinal segments of equal length (approximately<br />
10 cm each), beginning at the pylorus<br />
and ending at the ileocecal valve, and in the<br />
combined cecum—large intestine were counted.<br />
Empty cysts were seen but not counted in hosts<br />
necropsied at 1 and 2 hr p.i. The numbers were<br />
converted to percentages and the information is<br />
presented in Table 1. In Group A, the greatest<br />
percentage of encysted metacercariae was in<br />
segment 1 of the small intestine, and excysted<br />
metacercariae were recovered as far posteriad as<br />
segment 4 of the small intestine. Most of the<br />
4<br />
0.4<br />
0.5<br />
0.5<br />
1.7<br />
0.1<br />
2.0<br />
0<br />
1.7<br />
0<br />
2.2<br />
Copyright © 2011, The Helminthological Society of Washington<br />
5<br />
0.4<br />
0<br />
0.2<br />
0.6<br />
0.1<br />
0.3<br />
0<br />
0.4<br />
0<br />
0.5<br />
Cecumlarge<br />
intestine<br />
0.2<br />
0<br />
0.1<br />
0<br />
0.1<br />
0<br />
0<br />
0<br />
0<br />
0<br />
Total<br />
5.4<br />
7.5<br />
2.5<br />
8.6<br />
0.4<br />
11.4<br />
0<br />
10.1<br />
0<br />
10.3<br />
excysted metacercariae recovered at 1 hr p.i.<br />
were located in segment 3. Some excysted metacercariae<br />
were in the stomach at 1 hr and were<br />
alive and active. These organisms either had excysted<br />
in the stomach or, possibly, could have<br />
excysted in the small intestine and migrated anteriad<br />
to the stomach. In Group A, the finding<br />
of most excysted metacercariae in segments 1,<br />
2, and 3 of the small intestine (duodenum-jejunum<br />
region) provides support for claims that in<br />
vivo excystation takes place in the anterior part<br />
of the small intestine. This finding supports the<br />
statement of Simonsen et al. (1989) referenced<br />
above.<br />
With time, the ratio of encysted to excysted<br />
metacercariae declined (see last column in Table<br />
1), and by 4 hr p.i., encysted metacercariae were<br />
not found. These findings suggest that by 4 hr<br />
p.i. most of the encysted metacercariae had excysted<br />
or were voided. The idea of metacercariae<br />
being voided by 4 hr p.i. is consistent with<br />
the fact that the usual transit time for ingested<br />
food in the mouse digestive tract is 4 hr (Barrachina<br />
et al., 1997). Fecal examinations to determine<br />
the possible presence of excysted or encysted<br />
metacercariae in the stool were not made.<br />
About 75% of the excysted metacercariae in<br />
Groups C, D, and E were located in segments 1,<br />
2, and 3. Hence, newly excysted juveniles, up to<br />
at least 24 hr p.i., are more dispersed in the gut<br />
than are older worms. Manger and Fried (1993)<br />
showed that by day 2 p.i. more than 90% juvenile<br />
E. caproni were localized in segment 3 (the<br />
jejunum), and by day 4 and beyond, worms<br />
tended to migrate even more posteriad, with
most being found in segment 4 (jejunum-ileum<br />
zone).<br />
In conclusion, this study provides information<br />
on in vivo excystation of E. caproni metacercariae<br />
during the first day after infection in ICR<br />
mice. The in vivo studies show that most metacercariae<br />
excyst in the duodenum and migrate to<br />
the jejunum to become juveniles. The distribution<br />
of excysted metacercariae is quite variable<br />
during the first 24 hr of infection. The low recovery<br />
rates of organisms (only about 10%<br />
when mice received 100 cysts, and from 10.1%<br />
to 12.9% when mice received 400 cysts) attest<br />
to the fact that these organisms are difficult to<br />
detect in the first 24 hr after excystation. Perhaps<br />
some of these missing larvae are in sites other<br />
than the intestinal lumen, e.g., ducts or crypts<br />
associated with the intestine or other unknown<br />
locations. Manger and Fried (1993) did not report<br />
recovery percentages of juvenile worms of<br />
E. caproni from ICR mice at 2 d p.i. They did<br />
report a wide range of worm recoveries (18 to<br />
95%) from 2 to 8 d p.i. The fact that their recoveries<br />
were higher than the 10—13% in this<br />
study would suggest that some of the newly excysted<br />
metacercariae had been overlooked or<br />
had reemerged from extra-luminal sites.<br />
Support for this work was provided in part by<br />
funds from the Kreider Professorship to Dr. Ber-<br />
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 261-264<br />
Research Note<br />
RESEARCH NOTES 261<br />
nard Fried. We thank Ms. Vivienne R. Felix for<br />
typing the manuscript.<br />
Literature Cited<br />
Barrachina, M. D., V. Martinez, J. Y. Wei, and Y.<br />
Tache. 1997. Leptin-induced decrease in food intake<br />
is not associated with changes in gastric emptying<br />
in lean mice. American Journal of Physiology<br />
272:1007-1011.<br />
Fried, B. 1994. Metacercarial excystment of trematodes.<br />
Advances in <strong>Parasitology</strong> 33:91-144.<br />
, and S. Emili. 1988. Excystation in vitro of<br />
Echinostoma liei and E. revolutum (Trematoda)<br />
metacercariae. Journal of <strong>Parasitology</strong> 74:98-102.<br />
, and J. E. Huffman. 1996. The biology of the<br />
intestinal trematode Echinostoma caproni. Advances<br />
in <strong>Parasitology</strong> 38:311-3<strong>68</strong>.<br />
Hosier, D. W., and B. Fried. 1991. Infectivity,<br />
growth, and distribution of Echinostoma caproni<br />
(Trematoda) in the ICR mouse. Journal of <strong>Parasitology</strong><br />
77:640-642.<br />
Manger, P. M., Jr., and B. Fried. 1993. Infectivity,<br />
growth and distribution of preovigerous adults of<br />
Echinostoma caproni in ICR mice. Journal of Helminthology<br />
67:158-160.<br />
Simonsen, P. E., E. Bindseil, and M. K0ie. 1989.<br />
Echinostoma caproni in mice: studies on the attachment<br />
site of an intestinal trematode. International<br />
Journal for <strong>Parasitology</strong> 19:561-566.<br />
Ursone, R. L., and B. Fried. 1995. Light microscopic<br />
observations of Echinostoma caproni metacercariae<br />
during in vitro excystation. Journal of Helminthology<br />
69:253-257.<br />
Helminths Collected from Rattus spp. in Bac Ninh Province, Vietnam<br />
XUAN-DA PHAM,1'5 Cffl-LlEM TRAN,2 and HIDEO HASEGAWA3'4<br />
1 Department of Infectious Disease Control, Oita Medical University, Hasama, Oita 879-5593, Japan<br />
(e-mail: DA@oita-med.ac.jp),<br />
2 Ministry of Health, 138A Giang Vo, Ba Dinh, Hanoi, Vietnam, and<br />
3 Department of Biology, Oita Medical University, Hasama, Oita 879-5593, Japan<br />
(e-mail: hasegawa@oita-med.ac.jp)<br />
ABSTRACT: Helminthological examination was made<br />
on 35 rats (12 Rattus tanezumi, 14 Rattus argentiventer,<br />
and 9 Rattus losea) captured in 3 different habitats,<br />
i.e., residential, paddy field, and hilly areas, all in Bac<br />
Ninh Province, northern Vietnam. One trematode (Notocotylus<br />
sp.), 2 cestodes (Raillietina celebensis, Hymenolepis<br />
diminuta), 6 nematodes (Strongyloides ratti,<br />
Strongyloides venezuelensis, Nippostrongylus brasi-<br />
liensis, Orientostrongylus cf. tenorai, Syphacia muris,<br />
Gongylonema neoplasticum), and 1 acanthocephalan<br />
(Moniliformis monilifortnis) were collected. The species<br />
composition and prevalence of these helminths<br />
differed among the habitats, apparently because of biological<br />
characters of the parasites and environmental<br />
conditions of the localities.<br />
KEY WORDS: helminths, rat, Rattus tanezumi, Rattus<br />
Copyright © 2011, The Helminthological Society of Washington
262 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
argentiventer, Rattus losea, Trematoda, Notocotylus<br />
sp., Cestoda, Raillietina celebensis, Hymenolepis diminuta,<br />
Nematoda, Strongyloides ratti, Strongyloides venezuelensis,<br />
Nippostrongyhis brasiliensis, Orientostrongylus<br />
cf. tenorai, Syphacia muris, Gongylonema<br />
neoplasticum, Acanthoccphala, Moniliformis moniliformis,<br />
prevalence, ecology, Vietnam.<br />
There have been only limited reports on the<br />
parasites of rats from Vietnam (see Segal et al.,<br />
19<strong>68</strong>). Most previous surveys were carried out<br />
before 1970, and no data are available to assess<br />
the parasitological condition of rats at the present<br />
time. In 1999, we had an opportunity to<br />
examine helminths collected from rats trapped<br />
near Hanoi, northern Vietnam. Ten helminth<br />
species, including some of taxonomic and ecological<br />
interest, were found as recorded herein.<br />
Rats were collected with live traps in 3 different<br />
habitats, i.e., residential areas, paddy<br />
fields, and low hilly areas, all in Bac Ninh Province,<br />
Vietnam, in December 1999. They were<br />
anesthetized with ether and killed. Their viscera<br />
were fixed in 10% formalin solution and transported<br />
to the Oita Medical University, Oita, Japan.<br />
Their heads were also fixed in 10% formalin<br />
for species identification on the basis of<br />
skull morphology. On examination, the lung,<br />
heart, and liver were minced in water with fine<br />
forceps under a stereomicroscope to detect helminths<br />
parasitic in these organs. Then, the alimentary<br />
canal was cut open and washed on a<br />
stainless steel sieve with aperture size of 0.1<br />
mm. The residues left on the sieve were transferred<br />
to a Petri dish and observed under a stereomicroscope<br />
to recover helminths. The stomach<br />
wall was observed under a stereomicroscope<br />
with transillumination to find nematodes dwelling<br />
in the wall.<br />
Helminths collected were cleared in a glycerol-alcohol<br />
solution by evaporating alcohol<br />
and mounted on glass slides with a 50% glycerol<br />
solution. Some trematodes were stained<br />
with alum carmine or Heidenhain's iron hematoxylin,<br />
dehydrated in an alcohol series<br />
with ascending concentration, cleared in xylene<br />
and creosote, and mounted with Canada<br />
balsam. Voucher parasite specimens and host<br />
4 Corresponding author.<br />
5 Present address: Department of <strong>Parasitology</strong>, Hanoi<br />
Medical <strong>College</strong>, 1st Ton That Tung, Dong Da,<br />
Hanoi, Vietnam (e-mail: thuda<strong>2001</strong>@yahoo.com).<br />
Copyright © 2011, The Helminthological Society of Washington<br />
skulls are deposited in the National Science<br />
Museum, Tokyo (NSMT), Japan, with the accession<br />
numbers NSMT-P1 5073-5076,<br />
NSMT-As 2944-2953, and NSMT-M 31601-<br />
31606.<br />
The 35 rats examined included 12 black rats<br />
Rattus tanezumi Temminck, 1844 ( = so-called<br />
Asian-type Rattus rattus Linnaeus, 1758; cf.<br />
Musser and Carleton (1993)), 14 ricefield rats<br />
Rattus argentiventer (Robinson and Kloss,<br />
1916), and 9 lesser ricefield rats Rattus losea<br />
(Swinhoe, 1871). Helminths were not detected<br />
from the lung and liver, although Angiostrongylus<br />
cantonensis (Chen, 1935) and Calodium<br />
hepaticum (Bancroft, 1893) (syn. Capillaria<br />
hepatica (Bancroft, 1893)) have been previously<br />
recorded from those organs of rats in<br />
Vietnam (cf. Segal et al., 19<strong>68</strong>). Meanwhile,<br />
10 helminth species, comprising 1 trematode,<br />
2 cestodes, 6 nematodes, and 1 acanthocephalan,<br />
were collected from the alimentary canal<br />
(Table 1). Most of these helminths are common<br />
rat parasites, being widely distributed in<br />
the surrounding countries (Myers and Kuntz,<br />
1964, 1969; Ow-Yang, 1971; Singh and<br />
Cheong, 1971; Wiroreno, 1978; Sinniah, 1979;<br />
Ow-Yang and Durette-Desset, 1983; Hasegawa,<br />
1990; Hasegawa et al., 1992, 1994; Hasegawa<br />
and Syafruddin, 1995). Orientostrongylus<br />
sp. was found only in 2 rats from the<br />
paddy fields. Because no males were found,<br />
species identification is withheld, although it<br />
is strongly suggested to be Orientostrongylus<br />
tenorai Durette-Desset, 1970, a common rat<br />
parasite widely distributed from Afghanistan<br />
to Taiwan (Durette-Desset, 1970; Ow-Yang<br />
and Durette-Desset, 1983; Ohbayashi and Kamiya,<br />
1980; Hasegawa, 1990; Hasegawa et al.,<br />
1994; Hasegawa and Syafruddin, 1995).<br />
Among the parasites recovered, 2 cestodes,<br />
Hymenolepis diminuta (Rudolphi, 1819) and<br />
Raillietina celebensis (Janicki, 1902); 1 nematode,<br />
Gongylonema neoplasticum (Fibiger et<br />
Ditlevsen, 1914); and 1 acanthocephalan, Moniliformis<br />
monilifonnis (Bremser, 1811) have<br />
been recorded previously from Vietnamese<br />
rats (Segal et al., 19<strong>68</strong>). Notocotylus sp. is of<br />
special interest because trematodes of this genus<br />
have been reported only rarely from rats<br />
of the subfamily Murinae, although some<br />
members have often been recorded from voles<br />
of the subfamily Arvicolinae (cf. Yamaguti,
RESEARCH NOTES 263<br />
Table 1. Helminthic infections among rats collected from 3 different habitats in Bac Ninh Province,<br />
Vietnam.<br />
Rattus species:<br />
No. rats examined:<br />
Head and trunk length (cm) range:<br />
(Mean):<br />
Trematoda<br />
Notocotylus sp.<br />
Cestoda<br />
Raillietina celebensis<br />
Hymenolepis diminuta<br />
Nematoda<br />
Strongyloides spp.f<br />
Nippostrongylus brasiliensis<br />
Orientostrongylus cf. tenorai<br />
Syphacia murls<br />
Gongylonema neoplasticum<br />
Acanthocephala<br />
Moniliformis monilifonnis<br />
Habitat: Residential<br />
R. tanezumi<br />
10<br />
14-19<br />
(16.7)<br />
—<br />
2 (20)<br />
—<br />
—<br />
1 (10)<br />
—<br />
6 (60)<br />
4 (40)<br />
1 (10)<br />
* No. rats infected (prevalence in parentheses).<br />
t Strongyloides ratti and/or S. venezuelensis.<br />
1971). The morphological characteristics of<br />
Notocotylus sp. are summarized below.<br />
DESCRIPTION: Notocotylus sp. Trematoda:<br />
Notocotylidae. Body foliate, attenuated anteriorly,<br />
1.39-2.64 mm long by 0.57-0.91 mm<br />
wide; 3 longitudinal rows of prominent ventral<br />
glands present on ventral surface, each row<br />
composed of 12 to 14 glands; oral sucker terminal;<br />
esophagus short; ceca diverticulated,<br />
terminating just posterior to ovary; testes<br />
lobed, located lateral to terminal portion of<br />
ceca; genital pore immediately posterior to<br />
oral sucker; ovary intertesticular, lobed; vitellaria<br />
in lateral fields, extending from middle<br />
of body to anterior margins of testes; metraterm<br />
and cirrus sac almost equal in length; egg<br />
ellipsoidal, 22-24 by 11-12 |xm, with polar<br />
filaments, 2 to 3 times longer than egg length.<br />
HOSTS: Rattus argentiventer (Robinson et<br />
Kloss, 1916) and Rattus losea (Swinhoe, 1871).<br />
SITE IN HOSTS: Intestine.<br />
LOCALITY: Paddy field and low hilly area in<br />
Bac Ninh Province (21°7'N; 105°59'E), Vietnam.<br />
SPECIMENS DEPOSITED: National Science Museum,<br />
Tokyo, NSMT-P1 5073, 5074.<br />
REMARKS: In the neighboring areas of Vietnam,<br />
only 2 Notocotylus species have been<br />
recorded from mammals: Notocotylus mamii<br />
Hsu, 1954, of which adults were experimen-<br />
R. argentiventer<br />
1<br />
12-20<br />
(15.4)<br />
1 (14%)*<br />
—<br />
1 (14)<br />
2 (29)<br />
6 (86)<br />
1 (14)<br />
4 (57)<br />
—<br />
—<br />
Paddy field Low hilly area<br />
R. losea<br />
1<br />
10-18<br />
(14.2)<br />
2 (29)<br />
—<br />
—<br />
3 (43)<br />
3 (43)<br />
1 (14)<br />
3 (43)<br />
—<br />
R. tanezumi<br />
2<br />
17-17.5<br />
(17.3)<br />
—<br />
—<br />
—<br />
—<br />
2 (100)<br />
—<br />
2 (100)<br />
—<br />
R. argentiventer<br />
1<br />
16-21<br />
(17.5)<br />
3 (43)<br />
—<br />
—<br />
1 (14)<br />
7 (100)<br />
—<br />
2 (29)<br />
—<br />
•<br />
R. losea<br />
2<br />
13-14<br />
(13.5)<br />
1 (50)<br />
—<br />
—<br />
—<br />
2 (100)<br />
—<br />
2 (100)<br />
—<br />
tally raised in rabbits, and Notocotylus ratti<br />
Yie, Qiu, Weng, Li, and Li, 1956, the only<br />
representative exclusively known from Rattus,<br />
both from southern China (Hsu, 1954; Yie et<br />
al., 1956). Notocotylus mamii resembles the<br />
present form in the position of the genital pore<br />
but is readily distinguished in that the ceca are<br />
simple and the eggs have extremely elongated<br />
filaments, often more than 10 times longer<br />
than the egg length (Hsu, 1954, 1957). Although<br />
N. ratti resembles the present species<br />
in having diverticulated ceca, it is clearly distinguished<br />
by possessing a genital pore located<br />
posterior to the cecal bifurcation and only 5 to<br />
6 ventral glands in the median row (Yie et al.,<br />
1956). Presumably, the present worms represent<br />
a new species. However, proposal of a<br />
new taxon is withheld because the present<br />
worms were contracted by unsuitable fixation,<br />
obscuring some of the key structures.<br />
The species composition and prevalence of<br />
the rat helminths differed greatly among the<br />
habitats surveyed. Only Nippostrongylus brasiliensis<br />
(Travassos, 1914) and Syphacia muris<br />
(Yamaguti, 1935) were common to all 3 habitats.<br />
Raillietina celebensis, G. neoplasticum,<br />
and M. moniliformis were collected only from<br />
the rats captured around houses. Strongyloidid<br />
and trematode infections were not found in the<br />
rats captured around houses. Notocotylus sp.<br />
Copyright © 2011, The Helminthological Society of Washington<br />
—
264 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
was recovered from the rats collected in the<br />
paddy and hilly areas but not from those in the<br />
residential area.<br />
The differences in species composition and<br />
prevalence among the habitats could be explained<br />
by the biological characters of each<br />
helminth and by the environmental conditions.<br />
Because members of Notocotylus require a<br />
freshwater snail as the intermediate host (cf.<br />
Yamaguti, 1975), their presence in the paddy<br />
fields is reasonable. The hilly area surveyed in<br />
the present study contained many small ponds<br />
that might provide habitats for the snails. Also<br />
not unexpected is that G. neoplasticum and M.<br />
moniliformis were found only in the rats captured<br />
around the houses, because their intermediate<br />
hosts, usually cockroaches, are abundant<br />
in the residential areas. Nippostrongylus<br />
brasiliensis and Strongyloides spp. require<br />
moist soil for their embryonic and larval development<br />
and transmission. Their low prevalence<br />
or absence in the residential areas<br />
seems to be due to the dried soil around the<br />
houses in that season. Syphacia muris, the<br />
only helminth with relatively stable prevalence<br />
among the habitats, has a quite simple<br />
life cycle passed nearly entirely within the<br />
body of the host, and hence, its prevalence is<br />
less affected by the external environment.<br />
We are deeply indebted to Dr. G. G. Musser,<br />
American Museum of Natural History, for his<br />
kindness in identifying the rat species and to<br />
Dr. S. Kamegai, Meguro Parasitological Museum,<br />
for his kind consultation on Notocotylus<br />
sp.<br />
Literature Cited<br />
Durette-Desset, M. C. 1970. Caracteres primitifs de<br />
certains Nematodes Heligmosomes parasites de<br />
Murides et de Cricetides orientaux. Definition<br />
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 265-269<br />
Research Note<br />
RESEARCH NOTES 265<br />
Nematodes of the Tribe Cyathostominea (Strongylidae) Collected<br />
from Horses in Scotland<br />
J. RALPH LlCHTENFELS,1'5 AlOBHINN MCDONNELL,2 SANDY LOVE,3 AND<br />
JACQUELINE B. MATTHEWS4<br />
1 Biosystematics Unit, Parasite Biology, Epidemiology, and Systematics Laboratory, The Henry A. Wallace<br />
Beltsville Agricultural Research Center, Agricultural Research Service, U.S. Department of Agriculture,<br />
Beltsville, Maryland 20705-2350, U.S.A. (rlichten@anri.barc.usda.gov),<br />
2 Department of Veterinary <strong>Parasitology</strong>, University of Glasgow Veterinary School, Bearsden Road,<br />
Glasgow G61 1QH, United Kingdom,<br />
3 Weipers Centre for Equine Welfare, University of Glasgow Veterinary School, Bearsden Road,<br />
Glasgow G61 1QH, United Kingdom (S.Love@vet.gla.ac.uk), and<br />
4 Division of Equine Studies, Department of Veterinary Clinical Science and Animal Husbandry, Faculty of<br />
Veterinary Science, University of Liverpool, Leahurst CH64 7TE, United Kingdom (J.B.Matthews@liv.ac.uk)<br />
ABSTRACT: Nematodes of the tribe Cyathostominea<br />
are important parasites of horses. They live in large<br />
numbers in the large intestine and include over 50 species<br />
worldwide. This report describes an enumeration<br />
study of species found in a small population of horses<br />
in western Scotland. As found previously in a wide<br />
range of geographic regions, the 7 most abundant species<br />
of Cyathostominea, of the 18 recorded in this<br />
study, accounted for over 94% of the total population.<br />
One major exception to the results of previous studies<br />
was the presence of the most common species in this<br />
population, Cylicocyclus ashworthi. This species has<br />
not been recorded in the U.K. since its original description<br />
in 1924 and is morphologically very similar<br />
to another member of the same genus, Cylicocyclus<br />
nassatus, from which it has not been distinguished in<br />
previous studies in this geographical region. A rare<br />
species, Tridentoinfundibulum gobi, was found in low<br />
numbers in 3 of 4 horses.<br />
KEY WORDS: Nematoda, Cyathostominea, species<br />
survey, prevalence, intensity, horses, morphological<br />
identification, Scotland.<br />
Nematodes of the tribe Cyathostominea are<br />
the most common helminth parasites of the<br />
horse and are ubiquitous in all breeds. Members<br />
of the tribe Cyathostominea (Strongylidae) have<br />
been commonly referred to as small strongyles,<br />
cyathostomins (for the tribe), or cyathostomes<br />
(for the genus Cyathostomum Molin, 1861)<br />
(Hartwich, 1986). However, in order to avoid<br />
possible confusion with members of the nema-<br />
5 Corresponding author.<br />
tode genus Cyathostoma Blanchard, 1849 (Syngamidae),<br />
which are sometimes referred to as<br />
cyathostomes, we will use the common name<br />
cyathostomins to refer to the 51 species included<br />
in the tribe Cyathostominea as listed by Lichtenfels<br />
et al. (1998). Infections with these nematodes<br />
are complex: 51 species of cyathostomins<br />
have been recorded in horses, donkeys, and zebras<br />
worldwide (Lichtenfels et al., 1998), but 10<br />
of these species have been reported only from<br />
zebras or donkeys, and a few other species are<br />
known to have very limited distributions. However,<br />
most horses carry a burden of 5 to 10 common<br />
species, including many thousands (sometimes<br />
more than 100,000) of lumen-dwelling<br />
adult nematodes and as many larval stages in the<br />
walls of the large intestine (Reinemeyer et al.,<br />
1984; Bucknell et al., 1995). Clinically, cyathostomins<br />
are associated with various syndromes,<br />
the most dramatic of which is larval cyathostominosis,<br />
a fatal enteritis that occurs secondary to<br />
synchronized reactivation of arrested larvae<br />
from the intestinal mucosa (Giles et al., 1985;<br />
van Loon et al., 1995). The major obstacles to<br />
understanding, and therefore controlling, these<br />
parasites are their complexity, our inability to<br />
identify eggs in the feces, and the difficulty in<br />
identifying larvae on pasture. Until recently, the<br />
parasitic stages of cyathostomins could be identified<br />
only by adult worm morphology. However,<br />
recent studies have examined the molecular relationship<br />
of these species with a view to developing<br />
molecular probes for use in identifica-<br />
Copyright © 2011, The Helminthological Society of Washington
266 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
tion of both preparasitic and parasitic stages. For<br />
such studies, it is critical that the cyathostomins<br />
be identified and classified as consistently as<br />
possible. Modern identification manuals exist<br />
(Lichtenfels, 1975; Hartwich, 1986; Dvojnos<br />
and Kharchenko, 1994), but problems in identifying<br />
several species persist (Lichtenfels et al.,<br />
1997). In 1997, workers convened an international<br />
workshop to clarify the systematics of the<br />
Cyathostominea (Sun City, Republic of South<br />
Africa), and an agreement was reached on a consensus<br />
recommendation for the taxonomy of 51<br />
species as detailed in Lichtenfels et al. (1998).<br />
Despite the importance of these parasites, information<br />
is still lacking on species prevalence<br />
in Britain, especially since the development of<br />
widespread anthelmintic resistance. The last detailed<br />
study of species prevalence and infection<br />
intensity in the U.K. was in 1976 (Ogbourne,<br />
1976). The present report describes the species<br />
of cyathostomins present in the large intestine of<br />
a population of ponies from western Scotland.<br />
The nematodes were collected to provide DNA<br />
sequence information for the development of diagnostic<br />
tools and for phylogenetic analysis of<br />
the nematodes.<br />
Adult parasites were collected from intestinal<br />
contents of 4 Welsh-Shetland cross ponies aged<br />
from 9 to 15 mo originating from a local horse<br />
population. The history of anthelmintic treatment<br />
of the ponies is unknown. These animals<br />
were euthanized at the University of Glasgow<br />
Veterinary School for reasons other than parasite<br />
infestation. Intestinal contents were coarse-filtered<br />
with household plastic sieves. After sieving,<br />
the contents were passed through a Baermann<br />
apparatus with milk filters and then<br />
through SS-jjim wire-mesh sieves. Individual<br />
adult parasites were washed in sterile phosphatebuffered<br />
saline (137 mM NaCl, 8.1 mM<br />
Na2HPO4, 2.7 mM KC1, 1.47 mM KH2PO4, pH<br />
7.2). Where possible, a total of 200 parasites<br />
were collected from the ventral and dorsal colon;<br />
however, the cecum often contained fewer<br />
than 50 adult parasites. With the aid of a dissection<br />
microscope, the heads were excised with a<br />
scalpel because the bodies were subsequently<br />
used for DNA extraction. The heads were stored<br />
in 200 (JL! of 5% buffered formalin, then mounted<br />
on glass slides in a few drops of phenolalcohol<br />
(80% melted phenol crystals and 20%<br />
absolute ethanol) to which glycerine had been<br />
added at about 5% of the volume, and studied<br />
Copyright © 2011, The Helminthological Society of Washington<br />
with an Olympus BX50® differential interference<br />
contrast microscope. The parasites were<br />
identified according to the key of Lichtenfels<br />
(1975), supplemented by more recent descriptions<br />
of certain species (Lichtenfels and Klei,<br />
1988; Kharchenko et al., 1997; Lichtenfels et al.,<br />
1997, 1999). The taxonomy used in this report<br />
follows the checklist of genera and species recommended<br />
by the 1997 international workshop<br />
(Lichtenfels et al., 1998). Representative heads<br />
of 14 species of cyathostomins and Craterostomum<br />
acuticaudatum and photomicrographs of 2<br />
species of cyathostomins have been deposited in<br />
the U.S. National Parasite Collection, U.S. Department<br />
of Agriculture, Beltsville, Maryland<br />
20705-2350 as accession numbers 90698-<br />
90714. Two species of cyathostomins, Cylicocyclus<br />
elongatus and Cylicocyclus radiatus, and<br />
Gyalocephalus capitatus could not be documented<br />
by either method.<br />
Eighteen cyathostomin species, representing 5<br />
genera, were identified. Table 1 shows the total<br />
numbers of parasite species per animal identified<br />
morphologically and the relative numbers of<br />
each species collected from each animal. Eight<br />
species occurred in all 4 ponies. One rare species,<br />
Tridentoinfundibulum gobi, was found in<br />
•Scotland for the first time. It had been reported<br />
previously only in Asia and North America<br />
(Lichtenfels et al., 1998). In addition, individuals<br />
of the genera Craterostomum and Gyalocephalus<br />
were isolated but in smaller numbers<br />
than most of the cyathostomin species. The 7<br />
most abundant cyathostomin species were, in<br />
descending order, Cylicocyclus ashworthi, Cyathostomum<br />
catinatum, Cylicostephanus longibursatus,<br />
Cyclostephanus minutus, Cylicocyclus<br />
nassatus, Cylicocyclus insigne, and Cylicostephanus<br />
goldi. These species comprised over<br />
94% of the total cyathostomin burden. These results<br />
are similar to recent enumeration studies<br />
performed in several geographically distinct regions,<br />
for example in the U.S.A., Europe, and<br />
Australia (Reinemeyer et al., 1984; Mfitilodze<br />
and Hutchinson, 1985; Bucknell et al., 1995;<br />
Gawor, 1995). In addition, in terms of local studies<br />
performed previously in the U.K., the species<br />
identified here were very similar to those reported<br />
by Mathieson in Scotland (1964); Ogbourne<br />
in southwest England (1976), and Love<br />
and Duncan in Scotland (1992). Ogbourne<br />
(1976) performed the most extensive study and<br />
identified 21 species in 86 horses of various ages
RESEARCH NOTES 267<br />
Table 1. Numbers of specimens of nematodes, by species, collected from 4 ponies from western Scotland.<br />
Parasite species<br />
Cylicocyclus ashworthi (Le Roux, 1924) Mclntosh, 1933<br />
Cylicocycliis nassatus (Looss, 1900) Chaves, 1930<br />
Cylicocyclus insigne (Boulenger, 1917) Chaves, 1930<br />
Cylicocyclus ultrajectinus (Ihle, 1920) Ershov, 1939<br />
Cylicocyclus leptostomum (Kotlan, 1920) Chaves, 1930<br />
Cylicocyclus radiatus (Looss, 1900) Chaves, 1930<br />
Cylicocyclus elongatus (Looss, 1 900) Chaves 1 930<br />
Cyathostomum car ina turn Looss, 1900<br />
Cyathostomum pateratum (Yorke and Macfie, 1919) K'ung, 1964<br />
Coronocyclus coronatus (Looss, 1900) Hartwich, 1986<br />
Coronocyclus labiatus (Looss, 1900) Hartwich, 1986<br />
Cylicostephanus calicatus (Looss, 1900) Ihle, 1922<br />
Cylicostephanus longibitrsatiis (Yorke and Macfie, 1918) Cram, 1924<br />
Cylicostephanus minutiis (Yorke and Macfie, 1918) Cram, 1924<br />
Cylicostephanus goldi (Boulenger, 1917) Lichtenfels, 1975<br />
Cylicostephanus bidentatus (Ihle, 1925) Lichtenfels, 1975<br />
Cylidodontophoms bicoronatus (Looss, 1900) Ihle, 1922<br />
Tridentoinfundibulum gobi Tshoijo, in Popova, 1958<br />
Craterostomum acuticaudatum (Kotlan, 1919) Ihle, 1920<br />
Gyalocephalus capitatus Looss, 1900<br />
and breeds. In the latter study, 80% of these<br />
horses had C. longibursatus, C. goldi, C. calicatus,<br />
C. catinatum, C. coronatus, and C. nassatus.<br />
The most notable exception between the<br />
current study and all of these previous studies is<br />
the presence of C. ashworthi, the most prevalent<br />
species identified in our population. Cylicocyclus<br />
ashworthi was last reported in the U.K. as<br />
a new species (Le Roux, 1924) and has not been<br />
reported there since. Of note is that, in a comparable<br />
study performed several years earlier on<br />
worm populations derived from the same pastures<br />
as those used here, Love and Duncan<br />
(1992) identified 6 species, and C. nassatus was<br />
one of the most numerous. Cylicocyclus ashworthi<br />
and C. nassatus are morphologically very<br />
similar, and it is highly likely that these and other<br />
workers misidentified C. ashworthi as C. nassatus<br />
prior to the recent redescriptions of these<br />
species (Lichtenfels et al., 1997). Cylicocyclus<br />
nassatus is characterized by a cuticular shelf on<br />
the inner surface of the buccal capsule, a dorsal<br />
gutter that is as long as 50% of the buccal capsule<br />
depth, and 20 elements in the external leaf<br />
crown. Cylicocyclus ashworthi can be distinguished<br />
from C. nassatus by the absence of the<br />
shelf from the inner surface of the buccal capsule,<br />
by its much shorter dorsal gutter, and by<br />
25-29 external leaf crown elements that differ<br />
in shape from those of C. nassatus (Lichtenfels<br />
1<br />
94<br />
10<br />
24<br />
5<br />
0<br />
0<br />
0<br />
20<br />
9<br />
1<br />
0<br />
1<br />
24<br />
32<br />
26<br />
2<br />
0<br />
1<br />
4<br />
2<br />
2<br />
77<br />
51<br />
6<br />
1<br />
1<br />
0<br />
0<br />
51<br />
0<br />
2<br />
0<br />
2<br />
1 1<br />
55<br />
13<br />
4<br />
2<br />
1<br />
3<br />
0<br />
Pony<br />
3<br />
81<br />
47<br />
43<br />
1<br />
3<br />
1<br />
1<br />
80<br />
6<br />
0<br />
0<br />
3<br />
85<br />
6<br />
19<br />
9<br />
0<br />
3<br />
0<br />
0<br />
4<br />
83<br />
15<br />
14<br />
0<br />
0<br />
0<br />
0<br />
73<br />
0<br />
0<br />
1<br />
0<br />
23<br />
49<br />
4<br />
1<br />
0<br />
0<br />
1<br />
0<br />
et al., 1997). The ability to clearly observe the<br />
cuticular shelf in the buccal capsule is dependent<br />
on the clearing agent used, and this may have<br />
contributed to the difficulty in identifying this<br />
unique feature in previous studies.<br />
In addition to the historical difficulty in separating<br />
C. nassatus and C. ashworthi, C. ashworthi<br />
has also been misidentified as C. triramosus,<br />
which has also been confused with C.<br />
nassatus prior to its recent redescription (Kharchenko<br />
et al., 1997). We now know that C. triramosus<br />
is exclusively a parasite of zebras. It is<br />
imperative that C. nassatus and C. ashworthi be<br />
correctly differentiated because they are 2 of the<br />
most common nematodes found in the ventral<br />
colon of horses, and if DNA probes are to be<br />
developed on the basis of morphological delineation,<br />
then consistent identification is a prerequisite.<br />
Interestingly, Hung et al. (1997) performed<br />
sequencing of the first (ITS-1) and second<br />
(ITS-2) internal transcribed spacers of 5.8S<br />
ribosomal DNA of these species and found that<br />
C. nassatus and C. ashworthi, differentiated by<br />
head morphology, were sufficiently different at<br />
the DNA level to assign them to separate species.<br />
These results are similar to work performed<br />
on the intergenic spacer region of the nuclear<br />
DNA, where over 50% DNA sequence difference<br />
was found between these 2 species (Kaye<br />
et al., 1998), with low intraspecific variation<br />
Copyright © 2011, The Helminthological Society of Washington
2<strong>68</strong> COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
(0.3% for C. ashworthi and 1.9% for C. nassatus).<br />
Subsequently, oligoprobes designed from<br />
these IGS sequences have been used successfully<br />
to distinguish DNA of individual C. nassatus<br />
and C. ashworthi (Hodgkinson et al., <strong>2001</strong>). Furthermore,<br />
pairwise evolutionary distances, calculated<br />
under maximum likelihood with a GTR<br />
model estimated from data from the mitochondrial<br />
large ribosomal RNA subunit and ITS-2<br />
DNA, showed a 9.5% difference between the 2<br />
species (McDonnell et al., 2000).<br />
A consistent feature of all of the species incidence<br />
studies, including the present work, is<br />
that a small number of species, usually 5 to 10,<br />
constitute more than 80% of the infective load.<br />
Furthermore, the proportions of species have remained<br />
remarkably stable over many years, despite<br />
the widespread use of anthelmintics and<br />
the development of resistance. The rarer species<br />
are found at less consistent levels, but this is<br />
expected because the small populations may<br />
have gone undetected in cases where small subsamples<br />
of the worm populations have been examined<br />
for practical reasons. Consequently,<br />
much of the data on the least commonly discovered<br />
species are certain to underestimate true<br />
prevalence. Chapman et al. (1999) reported that<br />
9 to 15 species were found in a single animal<br />
when 200 specimens were identified, but the<br />
number increased to 20 to 29 when all nematodes<br />
in an entire 5% aliquot were identified.<br />
In the current study, Strongylus species were<br />
not found. This is probably indicative of the efficacy<br />
of anthelmintics and their strategic use in<br />
parasite control programs. In older studies, 100%<br />
prevalence of Strongylus species was reported<br />
(Le Roux, 1924; Foster and Ortiz, 1937). More<br />
recently, Bucknell et al. (1995) reported a prevalence<br />
of 38% Strongylus species in a study in<br />
which the deworming history of the horses was<br />
not known. Here, it was observed that, whereas<br />
relatively few species occurred exclusively in one<br />
or other parts of the intestines, most followed distinct<br />
site distributions (not shown), strongly biased<br />
in favor of a particular region, and (with the<br />
exception of C. ashworthi) these distributions<br />
were as described by Ogbourne (1976). Here, as<br />
in Ogbourne's study, the majority of C. pateraturn,<br />
C. insigne, and C. longibursatus individuals<br />
were found in the dorsal colon, whereas most of<br />
the C. nassatus, C. ultrajectinus, C. catinatum,<br />
and C. goldi adults were found in the ventral colon.<br />
Some caution must be taken in the interpre-<br />
Copyright © 2011, The Helminthological Society of Washington<br />
tation of these data because some of the species<br />
were found only in very low numbers. Also consistent<br />
with the findings of Ogbourne (1976) was<br />
that the cecum was the most sparsely populated<br />
region of the large intestine.<br />
This work was supported by a project grant<br />
funded by the Home of Rest for Horses near<br />
Lacey Green, Princes Risborough, Buckinghamshire,<br />
U.K. We thank James McGoldrick<br />
and colleagues at the Department of Veterinary<br />
<strong>Parasitology</strong>, University of Glasgow, for assistance<br />
in preparing the nematode heads. We<br />
thank Patrick Shone of Olympus for providing<br />
a microscope.<br />
Literature Cited<br />
Bucknell, D. G., R. B. Gasser, and I. Beveridge.<br />
1995. The prevalence and epidemiology of gastrointestinal<br />
parasites of horses in Victoria, Australia.<br />
International Journal for <strong>Parasitology</strong> 25:711-724.<br />
Chapman, M. R., D. D. French, and T. R. Klei.<br />
1999. Intestinal helminths of ponies; a comparison<br />
of species prevalent in Louisiana pre- and postivermectin.<br />
P. 74 in Proceedings of the American<br />
Association of Veterinary Parasitologists 44th Annual<br />
Meeting, New Orleans, Louisiana, July 10-<br />
13, 1999. (Abstract.)<br />
Dvojnos, G. M., and V. A. Kharchenko. 1994. Strongylidae<br />
in domestic and wild horses. Publishing<br />
House Naukova Dumka, Kiev, Ukraine. 234 pp.<br />
[In Russian.]<br />
Foster, A. O., and P. Ortiz O. 1937. A further report<br />
on the parasites of a selected group of equines in<br />
Panama. Journal of <strong>Parasitology</strong> 23:360-364.<br />
Gawor, J. J. 1995. The prevalence and abundance of<br />
internal parasites in working horses autopsied in<br />
Poland. Veterinary <strong>Parasitology</strong> 58:99-108.<br />
Giles, C. J., K. A. Urquhart, and J. A. Longstaffe.<br />
1985. Larval cyathostomiasis (immature trichonema-induced<br />
enteropathy): a report of 15 clinical<br />
cases. Equine Veterinary Journal 17:196-201.<br />
Hartwich, G. 1986. On the Strongylus tetracanthus<br />
problem and the systematics of the Cyathostominae<br />
(Nematoda, Strongyloidea). Mitteilungen aus<br />
dem Zoologischen Museum in Berlin 62:61-102.<br />
Hodgkinson, J. E., S. Love, J. R. Lichtenfels, S. Palfreman,<br />
Y. H. Ramsey, and J. B. Matthews.<br />
<strong>2001</strong>. Evaluation of the specificity of five oligoprobes<br />
for identification of Cyathostomin species<br />
from horses. International Journal for <strong>Parasitology</strong><br />
31:197-204.<br />
Hung, G. C., N. B. Chilton, I. Beveridge, A. Mc-<br />
Donnell, J. R. Lichtenfels, and R. B. Gasser.<br />
1997. Molecular delineation of Cylicocyclus nassatus<br />
and C. ashworthi (Nematoda: Strongylidae).<br />
International Journal for <strong>Parasitology</strong> 27:601-605.<br />
Kaye, J. N., S. Love, J. R. Lichtenfels, and J. B.<br />
McKeand. 1998. <strong>Comparative</strong> sequence analysis<br />
of the intergenic spacer region of cyathostome<br />
species. International Journal for <strong>Parasitology</strong> 28:<br />
831-836.
Kharchenko, V. A., G. M. Dvojnos, and J. R. Lichtenfels.<br />
1997. A redescription of Cylicocyclus triramosus<br />
(Nematoda: Strongyloidea): a parasite of<br />
the zebra, Equus burchelli antiquorum. Journal of<br />
<strong>Parasitology</strong> 83:922-926.<br />
Le Roux, P. L. 1924. Helminths collected from<br />
equines in Edinburgh and in London. Journal of<br />
Helminthology 2:111-134.<br />
Lichtenfels, J. R. 1975. Helminths of domestic equids.<br />
Illustrated keys to the genera and species with emphasis<br />
on North American forms. Proceedings of<br />
the Helminthological Society of Washington<br />
42(special issue): 1-92.<br />
, V. A. Kharchenko, R. C. Krecek, and L.<br />
M. Gibbons. 1998. An annotated checklist by genus<br />
and species of 93 species level names for 51<br />
recognized species of small strongyles (Nematoda:<br />
Strongyloidea: Cyathostominae) of horses, asses<br />
and zebras of the world. Veterinary <strong>Parasitology</strong><br />
79:65-79.<br />
-, C. Sommer, and M. Ito. 1997. Key<br />
characters for the microscopical identification of<br />
Cylicocyclus nassatus and Cylicocyclus ashworthi<br />
(Nematoda: Cyathostominea) of the horse, Equus<br />
caballus. Journal of the Helminthological Society<br />
of Washington 64:120-127.<br />
, and T. R. Klei. 1988. Cylicostephanus torbertae<br />
sp. n. (Nematoda: Strongyloidea) from Equus<br />
caballus with a discussion of the genera Cylicostephanus,<br />
Petrovinema and Skrjabinodentus.<br />
Proceedings of the Helminthological Society of<br />
Washington 55:165-170.<br />
, P. A. Pilitt, V. A. Kharchenko, and G. M.<br />
Dvojnos. 1999. Differentiation of Coronocychis<br />
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 269-272<br />
Research Note<br />
RESEARCH NOTES 269<br />
sagittatus and Coronocychis coronatus (Nematoda:<br />
Cyathostominea) of horses. Journal of the Helminthological<br />
Society of Washington 66:56-66.<br />
Love, S., and J. L. Duncan. 1992. Development of<br />
cyathostome infection of helminth naive foals.<br />
Equine Veterinary Journal Supplement 13:93-98.<br />
Mathieson, A. O. 1964. A study into the distribution<br />
of, and host tissue responses associated with,<br />
some internal parasites of the horse. Thesis, University<br />
of Edinburgh, Edinburgh, U.K. 160 pp.<br />
McDonnell, A., S. Love, A. Tait, J. R. Lichtenfels,<br />
and J. B. Matthews. 2000. Phylogenetic analysis<br />
of partial mitochondrial cytochrome oxidase C<br />
subunit I and large ribosomal RNA sequences and<br />
nuclear internal transcribed spacer I sequences<br />
from species of Cyathostominae and Strongylinae<br />
(Nematoda, Order Strongylida), parasites of the<br />
horse. <strong>Parasitology</strong> 121:649-659.<br />
Mfitilodze, M. W., and G. W. Hutchinson. 1985.<br />
Prevalence and abundance of equine strongyles<br />
(Nematoda: Strongyloidea) in tropical Australia.<br />
Journal of <strong>Parasitology</strong> 76:487-494.<br />
Ogbourne, C. P. 1976. The prevalence, relative abundance<br />
and site distribution of nematodes in the<br />
subfamily Cyathostominae in horses killed in Britain.<br />
Journal of Helminthology 50:203-214.<br />
Reinemeyer, C. R., S. A. Smith, A. A. Gabel, and<br />
R. P. Herd. 1984. The prevalence and intensity<br />
of internal parasites in horses in the USA. Veterinary<br />
<strong>Parasitology</strong> 15:75-83.<br />
van Loon, G., P. Deprez, E. Muylle, and B. Sustronck.<br />
1995. Larval cyathostomiasis as a cause<br />
of death in two regularly dewormed horses. Journal<br />
of Veterinary Medicine, Series A 42:301-306.<br />
Helminth Parasites of the Green Frog (Rana clamitans} from<br />
Southeastern Wisconsin, U.S.A.<br />
H. RANDALL YODER,' JAMES R. CoooiNS,2 AND J. CORBETT REINBOLD<br />
Department of Biological Sciences, University of Wisconsin-Milwaukee, P.O. Box 413, Milwaukee,<br />
Wisconsin 53201, U.S.A. (e-mail: 2coggins@uwm.edu)<br />
ABSTRACT: Between 13 August and 3 September 1999,<br />
26 green frogs Rana clamitans Rafinesque, 1820, were<br />
collected from 2 ponds at the University of Wisconsin-<br />
Milwaukee Field Station in Ozaukee County, Wisconsin,<br />
U.S.A. Hosts were euthanized and organs were examined<br />
for helminth parasites. All host individuals were infected<br />
1 Corresponding author. Current address: Department<br />
of Biology, Lamar University, P.O. Box 10037,<br />
Beaumont, Texas 77710, U.S.A. (e-mail:<br />
y oderhr @ hal .lamar.edu)<br />
with 1 or more helminth parasites. A total of 11 helminth<br />
species infected ./?. clamitans at this location: 9 platyhelminths<br />
(7 trematodes, 2 cestodes) and 2 nematodes. Mean<br />
abundance of infection was 65.5 ± 79.7 worms per host<br />
(range = 1-330). This is the first report of Clinostomum<br />
sp. from green frogs in Wisconsin.<br />
KEY WORDS: Amphibia, aquatic, Cestoda, Clinostomum,<br />
ephemeral pond, Haematoloechus varioplexus,<br />
green frog, helminth, Nematoda, parasites, Rana<br />
clamitans, survey, temporary pond, Trematoda, Wisconsin,<br />
U.S.A.<br />
Copyright © 2011, The Helminthological Society of Washington
270 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
The green frog Rana clamitans Rafmesque,<br />
1820, occurs from Newfoundland, where the<br />
population was introduced (Conant and Collins,<br />
1991), to western Ontario, Canada, in the northern<br />
extent of its range and from North Carolina<br />
to eastern Oklahoma, U.S.A. in the south (Vogt,<br />
1981). Although reports of green frog parasites<br />
are numerous, only 3 studies have been conducted<br />
in Wisconsin, U.S.A. (Williams and Taft,<br />
1980; Coggins and Sajdak, 1982; Bolek, 1998).<br />
A total of 26 green frogs were collected by hand<br />
between 13 August and 3 September 1999 from 2<br />
temporary ponds at the University of Wisconsin-<br />
Milwaukee Field Station, Ozaukee County, Wisconsin<br />
(43°23'N; 88°2'W). Frogs were transported<br />
to the laboratory and euthanized in MS-222 (ethyl<br />
m-aminobenzoate sulfonic acid). Body surface,<br />
mouth, eustachian tubes, celom, lungs, stomach,<br />
small intestine, colon, urinary bladder, liver, kidneys,<br />
and leg musculature in individual containers<br />
were examined with a dissecting microscope for<br />
the presence of helminth parasites. Nematodes<br />
were preserved in 70% ethanol and mounted in<br />
glycerin for identification. Larval and adult platyhelminths<br />
were fixed in alcohol-formalin-acetic<br />
acid, stained with acetic carmine, and mounted in<br />
Canada balsam. Voucher specimens were deposited<br />
at the H. W. Manter Helminth Collection, University<br />
of Nebraska, Lincoln, Nebraska (Table 1).<br />
Use of ecological terms follows the suggestions of<br />
Bush et al. (1997).<br />
All host individuals were infected with 1 or<br />
more helminths (prevalence = 100%). The component<br />
community of green frogs consisted of 11<br />
helminth species: 7 trematodes, 2 cestodes, and 2<br />
nematodes (Table 1). Overall mean abundance of<br />
helminths was 65.5 ± 79.7 worms per frog (range<br />
= 1—330). Haematoloechus varioplexus occurred<br />
with highest mean abundance, mean intensity, and<br />
prevalence of infection (Table 1). Nematodes occurred<br />
in low numbers and in few hosts (Table 1).<br />
Adult green frogs breed in a variety of permanent<br />
bodies of water (May-July in Wisconsin) and<br />
inhabit the periphery of these aquatic habitats<br />
throughout the summer (Vogt, 1981). During this<br />
time, adult frogs feed upon a variety of animals,<br />
including several species of insects with aquatic<br />
life histories (Jenssen and Klimstra, 1966). Whereas<br />
green frogs are known to migrate prior to hibernation,<br />
they are thought to seek out aquatic<br />
habitats that are well oxygenated and do not freeze<br />
entirely in winter (Lamoureux and Madison,<br />
1999). The ponds sampled in the present study are<br />
Copyright © 2011, The Helminthological Society of Washington<br />
ephemeral. Even in years when some water remains<br />
over winter, these ponds freeze solid. The<br />
green frogs that we collected seem to have moved<br />
into these ponds as a place to feed prior to hibernating<br />
in other areas.<br />
The species composition and numbers of helminths<br />
in green frog infracommunities at this location<br />
were similar to those reported previously<br />
(Rankin, 1945; Bouchard, 1951; Najarian, 1955;<br />
Campbell, 19<strong>68</strong>; Williams and Taft, 1980; Coggins<br />
and Sajdak, 1982; Muzzall, 1991; McAlpine,<br />
1997; Bolek, 1998; McAlpine and Burt, 1998).<br />
The aquatic habitat and diet of green frogs correspond<br />
with helminth communities consisting mostly<br />
of platyhelminths with indirect life cycles and<br />
relatively few direct life cycle nematodes. In the<br />
present study, H. varioplexus occurred with the<br />
highest values of prevalence, mean intensity, and<br />
mean abundance. These values are also high compared<br />
with those reported in previous studies.<br />
Muzzall (1991) reported 57% of 120 green frogs<br />
infected with H. parviplexus, synonymous with H.<br />
varioplexus (Kennedy, 1981), with a mean intensity<br />
of 29. Najarian (1955) reported 48% of 40<br />
green frogs infected with H. parviplexus and 42%<br />
prevalence for H. breviplexus but did not provide<br />
values for intensity or abundance of infection. Bolek<br />
(1998) reported a prevalence of 44% for H.<br />
varioplexus from 75 green frogs with a mean intensity<br />
of 5.3. Others have reported prevalence values<br />
of 25% or less for Haematoloechus spp. from<br />
R. clamitans (Rankin, 1945; Bouchard, 1951;<br />
Campbell, 19<strong>68</strong>; Williams and Taft, 1980; Mc-<br />
Alpine and Burt, 1998). Haematoloechus varioplexus<br />
has been reported previously from wood<br />
frogs (Rana sylvatica Le Conte, 1825) and spring<br />
peepers (Pseudacris crucifer Wied, 1839) from the<br />
same ponds sampled in the current study (Yoder<br />
and Coggins, 1996). It is therefore likely that infected<br />
intermediate hosts are present in these<br />
ponds. Additionally, large numbers of immature H.<br />
varioplexus were recovered from green frogs, indicating<br />
that hosts are being infected while feeding<br />
at these locations. Odonates serve as second intermediate<br />
hosts for species of Haematoloechus.<br />
Muzzall (1991) reported that the absence of fish<br />
predators may have increased the number of adult<br />
odonates emerging from Turkey Marsh, Michigan,<br />
U.S.A., resulting in richer helminth communities<br />
than those occurring in habitats where both frogs<br />
and fish occur. The absence of fish from these<br />
ephemeral ponds may have had a similar result in<br />
terms of high values of parasitism by H. vario-
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Stafford, 1905 (HWI<br />
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sstoda<br />
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ematoda<br />
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272 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
plexus. This is the first report of Clinostomum sp.<br />
from Wisconsin green frogs.<br />
We thank the U.W.M. Field Station Committee<br />
and staff for their support of this project.<br />
Literature Cited<br />
Bolek, M. G. 1998. A seasonal comparative study of<br />
helminth parasites in nine Wisconsin amphibians.<br />
M.S. Thesis, University of Wisconsin-Milwaukee,<br />
Milwaukee, Wisconsin, U.S.A. 134 pp.<br />
Bouchard, J. L. 1951. The Platyhelminthes parasitizing<br />
some northern Maine Amphibia. Transactions<br />
of the American Microscopical Society 70:<br />
245-250.<br />
Bush, A. O., K. D. Lafferty, J. M. Lotz, and A. W.<br />
Shostak. 1997. <strong>Parasitology</strong> meets ecology on its<br />
own terms: Margolis et al. revisited. Journal of<br />
<strong>Parasitology</strong> 83:575-583.<br />
Campbell, R. A. 19<strong>68</strong>. A comparative study of the<br />
parasites of certain Salientia from Pocahontas<br />
<strong>State</strong> Park, Virginia. Virginia Journal of Science<br />
19:13-20.<br />
Coggins, J. R., and R. A. Sajdak. 1982. Survey of<br />
helminth parasites in the salamanders and certain<br />
anurans from Wisconsin. Proceedings of the Helminthological<br />
Society of Washington 49:99-100.<br />
Conant, R., and J. T. Collins. 1991. A Field Guide<br />
to Reptiles and Amphibians: Eastern and Central<br />
North America. Houghton Mifflin Company, Boston,<br />
Massachusetts, U.S.A. 450 pp.<br />
Jenssen, T. A., and W. D. Klimstra. 1966. Food habits<br />
of the green frog, Rana clamitans, in southern<br />
Illinois. American Midland Naturalist 76:169-<br />
182.<br />
Kennedy, M. J. 1981. A revision of the genus Haematoloechus<br />
Looss, 1899 (Trematoda: Haemato-<br />
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 272-274<br />
Research Note<br />
loechidae) from Canada and the United <strong>State</strong>s.<br />
Canadian Journal of Zoology 59:1836-1846.<br />
Lamoureux, V. S., and D. M. Madison. 1999. Overwintering<br />
habitats of radio-implanted green frogs,<br />
Rana clamitans. Journal of Herpetology 33:430-<br />
435.<br />
McAlpine, D. F. 1997. Helminth communities in bullfrogs<br />
(Rana catesbeiand), green frogs (Rana<br />
clamitans), and leopard frogs (Rana pipiens) from<br />
New Brunswick, Canada. Canadian Journal of Zoology<br />
75:1883-1890.<br />
, and M. D. B. Burt. 1998. Helminths of bullfrogs,<br />
Rana catesbeiana, green frogs, R. clamitans,<br />
and leopard frogs, R. pipiens in New Brunswick.<br />
Canadian Field-Naturalist 112:50-<strong>68</strong>.<br />
Muzzall, P. M. 1991. Helminth infracommunities of<br />
the frogs Rana catesbeiana and Rana clamitans<br />
from Turkey Marsh, Michigan. Journal of <strong>Parasitology</strong><br />
77:366-371.<br />
Najarian, H. H. 1955. Trematodes parasitic in the Salientia<br />
in the vicinity of Ann Arbor, Michigan.<br />
American Midland Naturalist 55:195-197.<br />
Rankin, J. L. 1945. An ecological study of the helminth<br />
parasites of amphibians and reptiles of<br />
western Massachusetts and vicinity. Journal of<br />
<strong>Parasitology</strong> 31:142-150.<br />
Vogt, R. C. 1981. Natural History of Amphibians and<br />
Reptiles of Wisconsin. Milwaukee Public Museum<br />
and Friends of the Museum, Milwaukee, Wisconsin,<br />
U.S.A. 205 pp.<br />
Williams, D. D., and S. J. Taft. 1980. Helminths of<br />
anurans from NW Wisconsin. Proceedings of the<br />
Helminthological Society of Washington 47:278.<br />
Yoder, H. R., and J. R. Coggins. 1996. Helminth<br />
communities in the northern spring peeper, Pseudacris<br />
c. crucifer Weid, and the wood frog, Rana<br />
sylvatica Le Conte, from southeastern Wisconsin.<br />
Journal of the Helminthological Society of Washington<br />
63:211-214.<br />
Gastrointestinal Helminths of Spinner Dolphins Stenella longirostris<br />
(Gray, 1828) (Cetacea: Delphinidae) Stranded in La Paz Bay,<br />
Baja California Sur, Mexico<br />
ROGELIO AcuiLAR-AouiLAR,1 ROSA GRISELDA MORENO-NAVARRETE, GUILLERMO SALGADO-<br />
MALDONADO, and BERNARDO VILLA-RAMIREZ<br />
Universidad Nacional Autonoma de Mexico, Institute de Biologia, Apartado Postal 70-153, Mexico, D. F,<br />
CP 04510, Mexico<br />
ABSTRACT: Thirty-one spinner dolphins Stenella longirostris<br />
stranded in La Paz Bay, Baja California Sur,<br />
Mexico, were examined for endoparasitic helminths.<br />
The following species were identified: Zalophotrema<br />
Copyright © 2011, The Helminthological Society of Washington<br />
pacificum and Hadwenius tursionis (Digenea); Strobilocephalus<br />
triangularis, Trigonocotyle sp., and Tetraphyllidea<br />
gen. sp. larva (Cestoda); immature Bolbosoma<br />
hamiltoni (Acanthocephala); and Anisakis typica
(Nematoda). Except for H. tursionis, all the identified<br />
helminths are reported for the first time in Mexico.<br />
Stenella longirostris represents a new host for H. tursionis<br />
and A. typica.<br />
KEY WORDS: Cetacea, spinner dolphin, Stenella<br />
longirostris, parasites, Digenea, Cestoda, Nematoda,<br />
Acanthocephala, Gulf of California, Mexico.<br />
Although cetaceans, including dolphins, are<br />
common in marine waters of Mexico, their parasite<br />
fauna is poorly known. To date, only 2 reports<br />
on the helminth parasites of cetaceans in<br />
Mexico have been published. Lamothe-Argumedo<br />
(1987) identified the trematode Hadwenius<br />
tursionis (Marchi, 1873) in the intestine of<br />
the vaquita Phocoena sinus Norris and Mc-<br />
Farland, 1958 (Phocoenidae), from the northern<br />
Gulf of California, and Morales-Vela and Olivera-G6mez<br />
(1993) reported the trematode Nasitrema<br />
globicephala Neiland, Rice, and Holden,<br />
1970, and the nematodes Stenurus globicephalae<br />
Baylis and Daubney, 1925, Stenurus minor<br />
(Kuhn, 1829), and Crassicauda sp. in the pilot<br />
whale Globicephala macrorhynchus Gray, 1846<br />
(Delphinidae), from Cozumel Island, Quintana<br />
Roo (Caribbean Sea). The present report provides<br />
data on helminth occurrence in spinner<br />
dolphins Stenella longirostris (Gray, 1828) from<br />
the state of Baja California Sur, Mexico.<br />
In August 1993, 31 spinner dolphins were<br />
stranded in La Paz Bay (24°07'-24°21'N;<br />
110°17'-110°40'W), 20 km SW of the city of La<br />
Paz, Baja California Sur. The stranded dolphins<br />
consisted of 17 males (total length 130-188 cm,<br />
weight 19—57 kg, ages 1-18 yr) and 14 females<br />
(161-186 cm, 33-45 kg, 5.5-15 yr). The animals<br />
died of unknown causes during the strand,<br />
and they were kept deep frozen ( —22°C) until<br />
examination. During necropsy, the digestive<br />
tract of each animal was separated from its other<br />
viscera and examined for parasites. Trematodes<br />
and cestodes were fixed with Bouin's fluid and<br />
preserved in 70% ethanol, and acanthocephalans<br />
and nematodes were fixed and preserved in 70%<br />
ethanol. All helminths identified during the examination<br />
have been deposited in the Coleccion<br />
Nacional de Helmintos (National Helminth Collection)<br />
(CNHE) of the Universidad Nacional<br />
Autonoma de Mexico.<br />
Seven helminth species were recovered from<br />
1 Corresponding author (e-mail:<br />
raguilar@mail.ibiologia.unam.mx).<br />
RESEARCH NOTES 273<br />
the 31 dolphins. These include 2 trematodes:<br />
Zalophotrema pacificum Dailey and Perrin, 1973<br />
(bile ducts, prevalence 19%, mean intensity 6<br />
worms per parasitized host, range 1—16, CNHE<br />
No. 4018) and Hadwenius tursionis (Marchi,<br />
1873) (intestine, 6%, 1, 1-1, CNHE No. 4017);<br />
3 cestodes: Strobilocephalus triangularis (Diesing,<br />
1850) (rectum, 6%, 2, 2-2, CNHE No.<br />
4019), Trigonocotyle sp. (intestine, 90%, 5, 1-<br />
27, CNHE No. 4021; the poor condition of specimens<br />
preclude identification of species), and<br />
larval stages of Tetraphyllidea (intestine, 16%,<br />
31, 5-69, CNHE No. 4020); the nematode Anisakis<br />
typica (Diesing, 1860) (stomach, 77%, 18,<br />
1-98, CNHE No. 4023); and the immature acanthocephalan<br />
Bolbosoma hamiltoni Baylis, 1929<br />
(posterior intestine, 51%, 4, 1-9, CNHE No.<br />
4022).<br />
The helminth parasites of 5. longirostris have<br />
been reported by Delyamure (1955), Dailey and<br />
Brownell (1972), and Dailey and Perrin (1973).<br />
The previously recorded helminth fauna for this<br />
dolphin species includes the following: the trematodes<br />
Oschmarinella laevicaecum (Yamaguti,<br />
1942), Campula rochebruni (Poirier, 1886), Delphinicola<br />
tenuis Yamaguti, 1933, Lecithodesmus<br />
nipponicus Yamaguti, 1942, and Z. pacificum;<br />
the cestodes Diphyllobothrium fuhrmanni Hsu,<br />
1935, S. triangularis, Tetrabothrium forsteri<br />
(Krefft, 1871), Phyllobothrium delphini (Bosc,<br />
1802), Phyllobothrium sp., Monorygma grymaldii<br />
(Moniez, 1881), and Monorygma sp.; the<br />
nematodes Anisakis simplex (Rudolphi, 1809),<br />
Halocercus delphini Baylis and Daubney, 1925,<br />
and Mastigonema stenellae Dailey and Perrin,<br />
1973; and the acanthocephalans Bolbosoma vasculosum<br />
(Rudolphi, 1819), Bolbosoma balaenae<br />
(Gmelin, 1790), and Corynosoma sp. In the<br />
present study, the previously recorded Z. pacificum<br />
and S. triangularis are identified, and new<br />
host records are reported for H. tursionis, Trigonocotyle<br />
sp., tetraphyllidean B. hamiltoni, and<br />
A. typica. All but 1 of the identified species (H.<br />
tursionis) are recorded for the first time in Mexico.<br />
We thank Dr. Luis Fleischer and Hector<br />
Perez-Cortes, Centro Regional de Investigacion<br />
Pesquera, La Paz, Baja California Sur, for permission<br />
to examine dolphins; Dr. Tomas Scholz<br />
for confirmation of cestodes; Dr. Brent Nickol<br />
for review of the manuscript; Dr. Krzysztof<br />
Zdzitowiecki for providing helpful comments<br />
about the acanthocephalan; and Alejandro San-<br />
Copyright © 2011, The Helminthological Society of Washington
274 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
chez-Rios, Alejandra Nieto, and Francisco Anguiano<br />
for technical assistance. Collection of cetaceans<br />
was permitted by the Secretaria de Pesca,<br />
Mexico (authorization number 2275).<br />
Literature Cited<br />
Dailey, M. D., and R. L. Brownell. 1972. A checklist<br />
of marine mammal parasites. Pages 528-589 in S.<br />
H. Ridgeway, ed. Mammals of the Sea: Biology<br />
and Medicine. Charles C. Thomas, Springfield, Illinois,<br />
U.S.A.<br />
, and W. F. Perrin. 1973. Helminth parasites<br />
of porpoises of the genus Stenella in the eastern<br />
tropical Pacific, with description of two new species:<br />
Mastigonema stenellae gen. et sp. n. (Nematoda:<br />
Spiruroidea) and Zalophotrerna pacificurn<br />
n. sp. (Trematoda: Digenea). Fishery Bulletin 71:<br />
455-471.<br />
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 274-276<br />
Research Note<br />
Delyamure, S. L. 1955. The Helminth Fauna of Marine<br />
Mammals. Ecology and Phylogeny. Izdatel'stov<br />
Akademii Nauk SSSR. Translated 19<strong>68</strong>,<br />
Israel Program for Scientific Translations, Jerusalem,<br />
Israel. 522 pp.<br />
Lamothe-Argumedo, R. 1987. Trematodos de mamfferos<br />
III. Hallazgo de Synthesium tursionis (Marchi,<br />
1873) Stunkard y Alvey, 1930 en Phocoena<br />
sinus (Phocoenidae) en el Golfo de California,<br />
Mexico. Anales del Instituto de Biologia, Universidad<br />
Nacional Autonoma de Mexico, Serie<br />
Zoologia 58:11-20.<br />
Morales-Vela, D., and L. D. Olivera-Gomez. 1993.<br />
Varamiento de calderones Globicephala macrorhynchus<br />
(Cetacea: Delphinidae) en la Isla de<br />
Cozumel, Quintana Roo, Mexico. Anales del Instituto<br />
de Biologia, Universidad Nacional Autonoma<br />
de Mexico, Serie Zoologia 64:177-180.<br />
The Lung Nematodes (Metastrongyloidea) of the Virginia Opossum<br />
Didelphis virginiana in Southern California, U.S.A.<br />
VICTORIA E. MATEY,M BORIS I. KUPERMAN,' JOHN M. KiNSELLA,2 G. F. LLOYD,' AND<br />
S. J. LANE3<br />
1 Department of Biology and Center for Inland Waters, San Diego <strong>State</strong> University, San Diego, California 92182,<br />
U.S.A. (e-mail: kuperman@sunstroke.sdsu.edu),<br />
2 Department of Pathobiology, <strong>College</strong> of Veterinary Medicine, University of Florida, Gainesville, Florida 32611,<br />
U.S.A. (e-mail: wormdwb@aol.com), and<br />
3 Project Wildlife, P.O. Box 80696, San Diego, California 92138, U.S.A (e-mail: sjlane7@home.com)<br />
ABSTRACT: The lungworm Heterostrongylus heterostrongylus<br />
(Nematoda: Metastrongyloidea) is reported<br />
for the first time from the Virginia opossum Didelphis<br />
virginiana in North America. Seventeen of 31 opossums<br />
(55%) examined from San Diego County, California,<br />
U.S.A., were infected with H. heterostrongylus,<br />
with intensities ranging from 8 to 128 worms per host<br />
(mean 41). Another species of metastrongyloid nematode,<br />
Didelphostrongylus hayesi, was found in 74% of<br />
the lungs examined, with intensity ranging from 2 to<br />
1,328 worms per host (mean 312).<br />
KEY WORDS: lungworm, Heterostrongylus heterostrongylus,<br />
Nematoda, opossum, Didelphis virginiana,<br />
Didelphostrongylus hayesi, California, U.S.A.<br />
The Virginia opossum Didelphis virginiana<br />
Ken; 1792, is the only marsupial inhabiting<br />
Corresponding author.<br />
Copyright © 2011, The Helminthological Society of Washington<br />
North America, occurring in tropical, subtropical,<br />
and temperate habitats from southern Canada<br />
to Costa Rica (Gardner, 1973). California,<br />
U.S.A., was outside the original range of D. virginiana<br />
until its accidental introduction into Los<br />
Angeles County and the San Jose area from various<br />
eastern states between 1890 and 1910. By<br />
1958, D. virginiana was distributed widely in all<br />
the areas of California below 1,500 m altitude<br />
(Hunsaker, 1977).<br />
Until recently, the metastrongyloid lungworms<br />
of D. virginiana have been studied only<br />
in the midwestern and eastern U.S.A. Alden<br />
(1995) reviewed helminth records from the Virginia<br />
opossum and listed records of Didelphostrongylus<br />
hayesi Prestwood, 1976, from North<br />
Carolina, Georgia, Louisiana, and Tennessee,<br />
U.S.A. Subsequently, Baker et al. (1995) record-
RESEARCH NOTES 275<br />
Figure 1. Cephalic end of the lung nematode Heterostrongylus heterostrongylus from the Virginia opossum<br />
Didelphis virginiana. Male, frontal view. SEM. Am = amphid; Cl = collarette; Cp = cephalic papilla;<br />
L = lip; M = mouth opening; R = ring surrounding mouth. Scale bar = 20 u,m.<br />
ed D. hayesi from the Virginia opossum from<br />
Sacramento County in northern California. The<br />
objective of our study was to determine the identity<br />
and prevalence of lung parasites in feral Virginia<br />
opossums from southern California.<br />
Thirty-one Virginia opossums from San Diego<br />
County, California, killed by cars or euthanized<br />
after trauma, were examined for lung parasites<br />
from March 1999 to June 2000. All samples<br />
were obtained from a local nonprofit organization,<br />
Project Wildlife, Opossum Team,<br />
members of which also carried out the necropsy<br />
of the animals. All specimens were categorized,<br />
on the basis of weight, into juveniles (0.14-0.90<br />
kg) or adults (1.2-3.4 kg). The lungs with attached<br />
trachea of 7 juveniles and 24 adults were<br />
examined grossly and under the dissecting microscope.<br />
The trachea, bronchi, and bronchioles<br />
were split, and the lung parenchyma was teased<br />
apart gently. Worms recovered were fixed in 5%<br />
formalin or alcohol-formalin-acetic acid (AFA).<br />
For light microscopy, worms were examined as<br />
temporary whole mounts in glycerine after<br />
clearing in glycerine-alcohol with a Diastar®<br />
microscope equipped with a Photostar® camera<br />
system and were measured in micrometers. For<br />
scanning electron microscopy (SEM), the specimens<br />
were postfixed in 1% osmium tetroxide,<br />
followed by dehydration in an ethanol series,<br />
critical point dried with liquid CO2, sputter coated<br />
with gold-palladium, and examined with a<br />
Hitachi S-2700® scanning electron microscope.<br />
Voucher specimens of nematodes were deposited<br />
in the H. W. Manter Laboratory of <strong>Parasitology</strong>,<br />
University of Nebraska <strong>State</strong> Museum, Lincoln,<br />
Nebraska, U.S.A. (accession numbers<br />
15617-15619).<br />
In all, 7,381 lungworms were found in adult<br />
and juvenile animals examined. The parasites<br />
were identified as metastrongyloid nematodes.<br />
Of these, 91.1% were identified as D. hayesi and<br />
8.9% were identified as Heterostrongylus heterostrongylus<br />
Travassos, 1925. This is the first<br />
record of H. heterostrongylus from D. virginiana<br />
and the first record of this nematode in<br />
North America. Previous records of H. heterostrongylus<br />
were from another species of opossum,<br />
Didelphis marsupialis Linnaeus, 1758,<br />
Copyright © 2011, The Helminthological Society of Washington
276 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
from Colombia and Brazil, South America (Travassos,<br />
1925; Vaz and Pereira, 1934; Anderson<br />
et al., 1980).<br />
Morphologic and morphometric features of H.<br />
heterostrongylus in the opossums from southern<br />
California resembled those of specimens from<br />
South America described by Anderson et al.<br />
(1980). In D. virginiana, the male worms were<br />
slightly smaller, and the female worms were<br />
larger than in D. marsupialis. The mean length<br />
and width of H. heterostrongylus males from D.<br />
virginiana were correspondingly 6.5 mm (5.0—<br />
7.2) and 280 |xm (245-320). For females, the<br />
mean length was 9.7 mm (8.6-13.4) and the<br />
mean width was 380 jxm (350-475). SEM study<br />
showed some features of the cephalic structures<br />
that were not noted by Anderson et al. (1980).<br />
The 6 lips are completely fused, and each of<br />
them, in addition to 2 cephalic papillae, bears an<br />
amphid opening on the surface by an amphidial<br />
canal (Fig. 1). The shape of the mouth opening<br />
varies from triangular to circular (Fig. 1). A delicate<br />
ring surrounding the mouth opening and a<br />
collarette formed by a dilated cuticle are considered<br />
as permanent structures (Fig. 1). A new<br />
character of the female caudal extremity found<br />
in our study is a pair of small caudal papillae<br />
near the tip of the short blunt tail. The morphology<br />
of the male bursa, with its large lobe<br />
formed by the dorsal ray and short (93-100 |xm),<br />
complex and slightly arcuate spicules, is the<br />
same as previously reported.<br />
In the infected opossums, H. heterostrongylus<br />
were found lying freely in the bronchi. In 1 case,<br />
worms were recovered from the trachea.<br />
Seventeen of 31 opossums (55%) were infected<br />
with H. heterostrongylus, and intensities<br />
of infection ranged from 8 to 128 (mean 41).<br />
Infections were found in 58% of adult animals,<br />
with 12 to 128 worms per host (mean 41), and<br />
43% of juveniles, with intensities of 8 to 80<br />
worms per host (mean 44). The other species of<br />
lung nematodes, D. hayesi, was found under the<br />
pleura and was piercing lung tissue in 23 of 31<br />
opossums (74%). Intensities of infection ranged<br />
from 2 to 1,328 worms per host (mean 312). All<br />
animals infected by H. heterostrongylus were<br />
also infected by D. hayesi.<br />
Baker et al. (1995) reported on the prevalence<br />
and treatment of D. hayesi infections in opos-<br />
Copyright © 2011, The Helminthological Society of Washington<br />
sums collected in Yolo, Solano, and Sacramento<br />
counties in northern California. Infections were<br />
found in 23 of 33 opossums (70%). Because<br />
most of their infections were diagnosed by the<br />
presence of metastrongylid larvae in the feces<br />
and only 2 infections were confirmed by necropsies,<br />
it is possible that mixed infections of H.<br />
heterostrongylus and D. hayesi were also present<br />
in that area of California. Examination of<br />
additional host samples is desirable both in California<br />
and the eastern U.S.A. to determine the<br />
local range of H. heterostrongylus.<br />
We are deeply indebted to the Opossum Team<br />
of Project Wildlife, especially its leader, Blair<br />
Lee, and coleader, Beverly Rosas, and Meryl<br />
Faulkner, Mary Platter-Reiger, and numerous<br />
volunteers for collecting and providing specimens<br />
of the Virginia opossum for our studies.<br />
We are very grateful to D.V.M. Alfonso Guajardo,<br />
San Diego County Veterinarian Office, and<br />
Julie Irwin, student at San Diego <strong>State</strong> University<br />
(SDSU), for the necropsy of the opossums.<br />
Thanks are also due to SDSU student Susan<br />
Henderson for her technical assistance. We also<br />
thank 2 anonymous reviewers whose suggestions<br />
improved this paper.<br />
Literature Cited<br />
Alden, K. J. 1995. Helminths of the opossum, Didelphis<br />
virginiana, in southern Illinois, with a compilation<br />
of all helminths reported from this host in<br />
North America. Journal of the Helminthological<br />
Society of Washington 62:197-208.<br />
Anderson, R. C., M. D. Little, and U. R. Strelive.<br />
1980. The unique lungworms (Nematoda: Metastrongyloidea)<br />
of the opossum (Didelphis marsupialis<br />
Linnaeus). Systematic <strong>Parasitology</strong> 2:1-8.<br />
Baker, D. G., L. F. Cook, E. M. Johnson, and N.<br />
Lamberski. 1995. Prevalence, acquisition, and<br />
treatment of Didelphostrongylus hayesi (Nematoda:<br />
Metastrongyloidea) infection in opossums (Didelphis<br />
virginiana). Journal of Zoo and Wildlife<br />
Medicine 26:403-408.<br />
Gardner, A. L. 1973. The systematics of the genus<br />
Didelphis (Marsupialia: Didelphidae) in North and<br />
Middle America. Special Publications of the Museum,<br />
Texas Tech University 4:3-45.<br />
Hunsaker, D. 1977. Ecology of New World marsupials.<br />
Pages 95-156 in D. Hunsaker II, ed. The<br />
Biology of Marsupials. Academic Press, New<br />
York-San Francisco-London.<br />
Travassos, L. 1925. Un nouveau type de Metastrongylidae.<br />
Comptes Rendus des Seances de la Societe<br />
de Biologic, Paris 93:1259-1262.<br />
Vaz, Z., and C. Pereira. 1934. Some new Brazilian<br />
nematodes. Journal of the Washington Academy<br />
of Sciences 24:54.
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 277-282<br />
Research Note<br />
RESEARCH NOTES 277<br />
New Host and Distribution Records of Cosmocephalus obvelatus<br />
(Creplin, 1825) (Nematoda: Acuariidae), with Morphometric<br />
Comparisons<br />
JULIA I. DiAZ,1 GRACIELA T. NAVONE, AND FLORENCIA CREMONTE<br />
Centre de Estudios Parasitologicos y de Vectores (CONICET-UNLP), Calle 2 NQ 584, 1900 La Plata,<br />
Argentina (e-mail: cepave@netverk.com.ar)<br />
ABSTRACT: This is the first record of Cosmocephalus<br />
obvelatus (Creplin, 1825) Seurat, 1919 (Nematoda:<br />
Acuariidae), from Argentina (Valdes Peninsula, province<br />
of Chubut) and from the Magellanic penguin<br />
Spheniscus magellanicus (Aves: Spheniscidae). The<br />
prevalence of this parasite was 31.3% and the mean<br />
intensity was 5.4. Despite the wide geographic distribution<br />
and the great variety of hosts parasitized by C.<br />
obvelatus (14 families belonging to 8 orders), there<br />
were no significant differences in morphological characteristics<br />
and measurements from previous records.<br />
Both the wide distribution and the morphometrical stability<br />
of C. obvelatus may be explained by its ecology<br />
and mode of transmission.<br />
KEY WORDS: Cosmocephalus obvelatus, Acuariidae,<br />
Nematoda, Spheniscus magellanicus, Spheniscidae,<br />
marine birds, Argentina.<br />
Cosmocephalus obvelatus (Creplin, 1825)<br />
Seurat, 1919, an acuariid nematode with a wide<br />
distribution, has been previously reported in Europe,<br />
Asia, Africa, New Zealand, and North<br />
America (Wong and Anderson, 1982). In South<br />
America, there is only 1 record of C. obvelatus,<br />
described as Cosmocephalus tanakai by Rodrigues<br />
de Olivera and Vicente (1963) from the<br />
black-backed gull Larus dominicanus Lichtenstein,<br />
1823, in Brazil. Later, C. tanakai was synonymized<br />
with C. obvelatus by Anderson and<br />
Wong (1981). This parasite has a wide range of<br />
hosts, having been previously recorded in members<br />
of Lariidae, Pelecanidae, Rynchopidae,<br />
Sternidae, Anatidae, Podicipedidae, Phalacrocoracidae,<br />
Gaviidae, Ardeidae, Stercoraridae,<br />
Haematopodidae, Treschiornitidae, and Accipitridae<br />
(Barus and Majudmar, 1975; Borgsteede<br />
and Jansen, 1980; Anderson and Wong, 1981;<br />
1 Corresponding author (e-mail: jid027@yahoo.<br />
com.ar).<br />
Tuggle and Schmeling, 1982). Among members<br />
of the Spheniscidae, C. obvelatus has been cited<br />
only from the rockhopper penguin Eudyptes<br />
crestatus (Miller, 1784) caught in Chile and<br />
transferred to the Japanese Zoological Garden<br />
(Azuma et al., 1988).<br />
This note reports the first record of C. obvelatus<br />
in the Magellanic penguin Spheniscus magellanicus<br />
(Forster, 1781) (Aves: Spheniscidae).<br />
It is also the first time that C. obvelatus has been<br />
found in Argentina. Measurements of the specimens<br />
in this study are compared with those given<br />
by previous authors. Morphological details<br />
seen in the scanning electron microscope (SEM)<br />
and dates of prevalence and mean intensity are<br />
provided.<br />
At irregular intervals from 1996 to 2000, 16<br />
specimens of S. magellanicus, all of which had<br />
recently died of unknown but presumably natural<br />
causes, were collected along the coasts of the<br />
Valdes Peninsula (42°04'-42°53'S, 63°38'-<br />
64°30'W), province of Chubut, Argentina. After<br />
dissection, the digestive tract was fixed in 10%<br />
formalin. Acuariid nematodes were removed<br />
from the esophagus and stored in 70% ethanol.<br />
The specimens were cleared in lactophenol and<br />
studied under the light microscope. Some specimens<br />
were dried by the critical point method,<br />
examined by SEM (Jeol/SET 100®), and photographed.<br />
Voucher specimens were deposited in<br />
the Helminthological Collection of the Museo de<br />
La Plata (CHMLP), La Plata, Argentina (Accession<br />
no. 4811).<br />
The measurements of our specimens and<br />
those given by previous authors are listed in Table<br />
1. Morphological details are shown in Figures<br />
1-6. The prevalence was 31.25% and the<br />
mean intensity was 5.4. The esophagus was the<br />
only site of infection. We observed several de-<br />
Copyright © 2011, The Helminthological Society of Washington
Table 1. <strong>Comparative</strong> measurements of Cosmocephalus obvelatus from different hosts and localities.<br />
Rodrigues de<br />
Olivera and Anderson and Wong<br />
Vicente (1963) (1981)<br />
Cram (1927) Khalil (1931) Rao (1951)<br />
La<br />
Larus sp.<br />
Host* Several Pelecanus sp. Larus sp.<br />
Ne<br />
Larus delawarensis<br />
Canada<br />
Brazil<br />
Locality Europe Egypt Canada<br />
4<br />
10<br />
27<br />
—<br />
39<br />
45<br />
—<br />
—<br />
—<br />
3.3<br />
En<br />
44<br />
10<br />
19.4 (15.8-22.3)<br />
393 (320-500)<br />
615 (570-730)<br />
<strong>68</strong>4 (640-770)<br />
<strong>68</strong>5 (610-790)<br />
813 (705-940)<br />
1.3 (1.2-1.5)<br />
4.7 (4.1-5.1)<br />
6.0 (5.2-6.6)<br />
End of lateral alae<br />
8.4 (7.4-10.4)<br />
1<br />
12.5<br />
277<br />
363<br />
—<br />
—<br />
—<br />
0.77<br />
3.43<br />
4.2<br />
Female<br />
n — 1 immature —<br />
Total length (mm) 9.7-20 5.7 37122<br />
Maximum width (|xm) 300-380 — 200-400<br />
Buccal capsule ((Am) — — —<br />
Nerve ring (n-m) — — —<br />
Deirids (u.m) 490 — —<br />
Excretory pore (|xm) — — —<br />
Muscular esophagus (mm) — — —<br />
Glandular esophagus (mm) — — —<br />
Total esophagus (jxm) — <strong>68</strong>0 —<br />
Postdeirids — — —<br />
Vulva (from anterior end) (mm) 5.5 Midbody Midbody<br />
6.2<br />
—<br />
—<br />
39<br />
21<br />
—<br />
Long<br />
Short<br />
43 (40-45)<br />
25<br />
301 (220-380)<br />
—<br />
—<br />
36<br />
19<br />
175<br />
Vagina vera (pun) — — —<br />
Vagina uterina (|xm) — — —<br />
Egg length (p,m) 36 — 35-37<br />
Egg width (fi,m) 20 — 17-18<br />
Tail ((i,m) 230 180 —<br />
10<br />
10<br />
27<br />
—<br />
—<br />
46<br />
55<br />
—<br />
—<br />
5.2<br />
60<br />
10<br />
12.4 (9.9-14.3)<br />
279 (200-350)<br />
418 (380-510)<br />
474 (420-530)<br />
450 (350-540)<br />
583 (500-<strong>68</strong>0)<br />
1.1 (1.0-1.3)<br />
4.0 (3.6-4.3)<br />
5.1 (4.6-5.4)<br />
End of lateral alae<br />
2<br />
9.5-12<br />
250-280<br />
369<br />
462<br />
399<br />
532-630<br />
0.98-1.32<br />
2.85-4.61<br />
3.83-5.93<br />
—<br />
Male<br />
n — 2 —<br />
Total length (mm) 5.7-12.2 7.6 37085<br />
Maximum width (jjum) 240-255 — 150-270<br />
Buccal capsule (\j.m) — — —<br />
Nerve ring ((JUTI) — — —<br />
Deirids (p,m) 430 — —<br />
Excretory pore (u,m) — — —<br />
Muscular esophagus (mm) — — —<br />
Glandular esophagus (mm) — — —<br />
Total esophagus (|j,m) — 930 —<br />
Postdeirids — — —<br />
Copyright © 2011, The Helminthological Society of Washington<br />
COMP>
c<br />
Tj ;^ fn<br />
in<br />
ON<br />
o<br />
m<br />
ON<br />
"3<br />
s<br />
ON<br />
gcd<br />
u<br />
r? >ri ?<br />
— i r- r-<br />
7 i T<br />
CN r~ oc<br />
C- S C<br />
CN NO NC<br />
NO CN •r-i<br />
in
280 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Figures 1-6. Cosmocephalus obvelatus from Spheniscus magellanicus. 1. Apical view. 2. Anterior extremity<br />
showing lateral alae (arrow), dorsal view. 3. Anterior extremity showing cephalic papillae (arrow),<br />
inflexions of chordons and deirid (arrow), lateral view. 4. Detail of deirid. 5. Detail of postdeirid located<br />
near end of lateral alae (arrow). 6. Posterior extremity of male with left spicule protruded and showing<br />
papillae arrangement with proximal pair of precloacal papillae lying out of line of distribution (arrow),<br />
latero-ventral view. Scale bars: 1 = 50 ixm, 2 = 500 (mm, 3 = 100 u.m, 4 = 10 (xm, 5 = 20 (Jim, and 6 =<br />
200 fjim.<br />
Copyright © 2011, The Helminthological Society of Washington
ticeps (Rudolphi, 1819), which also is cosmopolitan<br />
but has a narrower range of hosts, Etchegoin<br />
et al. (2000) found differences in measurements<br />
among specimens from different localities.<br />
The shape of the cordons, the morphology and<br />
size of the cervical papillae, and the location in<br />
the definitive host (habitats) seem to be of fundamental<br />
importance when establishing relationships<br />
in the acuariid group (Barus and Majudmar,<br />
1975). The cordons of Cosmocephalus have<br />
a complex structure and are relatively wide.<br />
Cosmocephalus obvelatus is always located in<br />
the esophagus of the host. The genera Synhimantus<br />
and Cosmocephalus are closely related;<br />
they have similar cordons and cervical papillae,<br />
but members of the former genus live under the<br />
cuticle of the gizzard. Etchegoin et al. (2000)<br />
reported morphometric differences that they<br />
considered as intraspecific variations in specimens<br />
from different hosts and localities. However,<br />
we observed that C. obvelatus varies little,<br />
even in different hosts and localities. This morphometrical<br />
stability may indicate that Cosmocephalus<br />
is better adapted to different hosts and<br />
diverse localities because all hosts have similar<br />
environmental and feeding habits (eating fish).<br />
The intermediate hosts of C. obvelatus are amphipods,<br />
and it uses fish as paratenic hosts (Anderson,<br />
1992). These characteristics may play an<br />
important role in the cosmopolitan distribution<br />
of C. obvelatus. Moreover, the distributions of<br />
many species of fish-eating birds overlap in their<br />
breeding and/or wintering grounds.<br />
The intensity of infection recorded here is<br />
similar to the intensities found by Keppner<br />
(1973) from the California gull Larus californicus<br />
Lawrence, 1854 (Lariidae) (prevalence [P]<br />
= 23.5% and mean intensity [I] = 4.25), by<br />
Courtney and Forrester (1974) from the brown<br />
pelican Pelecanus occidentalis Linnaeus, 1776<br />
(Pelecanidae), in North America (P = 40% and<br />
I = 4), and by Lafuente et al. (1999) from Audouin's<br />
gull Larus audouinii Payraudeau, 1826<br />
(Lariidae), in the Mediterranean Sea (P =<br />
82.76% and I = 5.08). This intensity is higher<br />
than that given by Threlfall (19<strong>68</strong>) from the<br />
black-backed gull Larus marinus Linnaeus,<br />
1758 (Lariidae), in Newfoundland (P = 9.39%<br />
and I = 1).<br />
Boero and Led (1970) described a new species,<br />
Cosmocephalus argentinensis, from 1 female<br />
specimen found in a Magellanic penguin<br />
RESEARCH NOTES 281<br />
in the Zoological Garden in La Plata, Argentina.<br />
We consider this acuariid as a species inquirendae<br />
because the description is very poor and no<br />
type materials (which were never deposited in a<br />
museum collection) are available for our examination.<br />
We gratefully acknowledge the staff of the<br />
Servicio de Microscopia Electronica de Barrido,<br />
Museo de La Plata, for their technical assistance<br />
and Lucy Shirlaw for revision of the English.<br />
This study was funded by the Consejo Nacional<br />
de Investigaciones Cientfficas y Tecnicas (CON-<br />
ICET) and by the Comision de Investigaciones<br />
Cientfficas de la Provincia de Buenos Aires<br />
(CIC).<br />
Literature Cited<br />
Anderson, R. C. 1992. Nematode Parasites of Vertebrates.<br />
Their Development and Transmission.<br />
CAB International, Wallingford, Oxon, U.K. 578<br />
pp.<br />
, and P. L. Wong. 1981. Redescription of Cosmocephalus<br />
obvelatus (Creplin, 1825) (Nematoda:<br />
Acuarioidea) from Larus delawarensis Ord (Laridae).<br />
Canadian Journal of Zoology 59:1897-<br />
1902.<br />
Azuma, H., M. Okamoto, M. Ohbayashi, Y. Nishine,<br />
and T. Mukai. 1988. Cosmocephalus obvelatus<br />
(Creplin 1825) (Nematoda: Acuariidae) collected<br />
from esophagus of rockhopper penguin, Eudyptes<br />
crestatus. Japanese Journal of Veterinary<br />
Research 36:73-77.<br />
Barus, V., and G. Majudmar. 1975. Scanning microscopic<br />
studies on the cordon structures of Acuariid<br />
genera (Nematoda: Acuariidae). Folia Parasitologica<br />
22:125-131.<br />
Boero, J. J., and J. E. Led. 1970. El parasitismo de<br />
La Fauna Autoctona. VI. Los parasites de la avifauna<br />
argentina I. Actas de las 5a Jornadas de Veterinaria,<br />
Facultad de Ciencias Veterinarias, Universidad<br />
Nacional de La Plata: 65-71.<br />
Borgsteede, F. H. M., and J. Jansen. 1980. Spirurata<br />
in wild birds in The Netherlands. Tropical and<br />
Geographic Medicine 32:91-92.<br />
Bowie, J. Y. 1981. Redescription of Cosmocephalus<br />
tanakai Rodriguez and Vicente (Nematoda-Acuariidae)<br />
a parasite of the southern black-backed<br />
gull in New Zealand. New Zealand Journal of Zoology<br />
8:249-253.<br />
Courtney, C. H., and D. J. Forrester. 1974. Helminth<br />
parasites of the brown pelican in Florida<br />
and Louisiana. Proceedings of the Helminthological<br />
Society of Washington 41:89-93.<br />
Cram, E. B. 1927. Bird parasites of the suborders<br />
Strongylata, Ascaridata and Spirurata. Bulletin of<br />
the United <strong>State</strong>s National Museum 140:1-465.<br />
Etchegoin, J. A., F. Cremonte, and G. T. Navone.<br />
2000. Synhimantus (Synhimantus) laticeps (Rudolphi,<br />
1819) Railliet, Henry et Sisoff, 1912<br />
(Nematoda, Acuariidae) parasitic in Tyto alba<br />
Copyright © 2011, The Helminthological Society of Washington
282 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
(Gmelin) (Aves, Tytonidae) in Argentina. Acta<br />
Parasitologica 45:99-106.<br />
Keppner, E. J. 1973. Some parasites of California<br />
gull, Lams californicus Lawrence, in Wyoming.<br />
Transactions of the American Microscopical Society<br />
92:288-291.<br />
Khalil, M. B. 1931. On two new species of nematodes<br />
from Pelecanus onocrotalus. Annals of Tropical<br />
Medicine and <strong>Parasitology</strong> 25:455-460.<br />
Lafuente, M., V. Roca, and E. Carbonell. 1999. Cestodos<br />
y nematodos de la gaviota de Audouin, Lams<br />
audouinii Payraudeau, 1826 (Aves: Laridae)<br />
en las Islas Chafarinas (Mediterraneo sudoccidental).<br />
Boletin de la Real Sociedad Espanola de Historia<br />
Natural (Sec. Biol.) 95:13-20.<br />
Rao, N. 1951. Cosmocephalus firlottei n. sp. (family<br />
JOHN S. MACKIEWICZ<br />
Elected to Life Membership in the Helnlinthological<br />
Society of Washington November 15, 2000<br />
Copyright © 2011, The Helminthological Society of Washington<br />
Acuariidae) from the sea gull Larus argentatus.<br />
Canadian Journal of Zoology 25:173-177.<br />
Rodrigues de Olivera, H., and J. J. Vicente. 1963.<br />
Nova especie do genero Cosmocephalus Molin,<br />
1858 (Nematoda, Spiruroidea). Revista Brasilera<br />
de Biologia 23:389-392.<br />
Threlfall, W. 19<strong>68</strong>. The helminth parasites of three<br />
species of gulls in Newfoundland. Canadian Journal<br />
of Zoology 46:827-830.<br />
Tuggle, B. N., and S. K. Schmeling. 1982. Parasites<br />
of the bald eagle (Haliaetus leucocephalus) of<br />
North America. Journal of Wildlife Diseases 18:<br />
501-506.<br />
Wong, P. L., and R. C. Anderson. 1982. The transmission<br />
and development of Cosmocephalus obvelatus<br />
(Nematoda: Acuarioidea) of gulls (Laridae).<br />
Canadian Journal of Zoology 60:1426-1440.<br />
GRAHAM C. KEARN<br />
Elected to Life Membership in the Helminthological<br />
Society of Washington November 15, 2000
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 283-284<br />
Anniversary Award<br />
The Helminthological Society of Washington<br />
NANCY D. PACHECO<br />
Harley G. Sheffield, left, presents the 2000 Anniversary Award to Nancy D. Pacheco<br />
As Chairman of the Anniversary Award Committee of the Helminthological Society of Washington,<br />
it is my duty to present the 2000 Anniversary Award to Nancy Pacheco. Not only is it a<br />
duty, it is an honor and a pleasure to be able to present this award to such an outstanding member<br />
of the Society.<br />
The Award is authorized by the Society's Constitution and is to be given to a member for one<br />
or more achievements of the following nature: an outstanding contribution to the science of parasitology<br />
or related sciences that brings honor and credit to the Society, an exceptional paper read<br />
at a meeting of the Society or published in the Society's journal, outstanding service to the Society,<br />
or another achievement or contribution of distinction that warrants the highest recognition by the<br />
Society. The Awards Committee determined that Nancy qualifies in all of the above categories.<br />
Nancy was born and educated in Kansas. During her junior year of high school, she was fortunate<br />
to be an exchange student under the American Field Service Program and lived 6 months in New<br />
Zealand. She subsequently received the Bachelor of Science Degree from Washburn University in<br />
Topeka. Following graduation, she came to the National Institutes of Health and received a position<br />
as a biologist in the National Heart Institute. Working with physicians and postdocs, Nancy was<br />
engaged in studies on cardiac muscle physiology. Maybe there was a lot of twitching in that job<br />
because for some reason, she saw the light and soon turned to parasitology. In 19<strong>68</strong>, and for the<br />
283<br />
Copyright © 2011, The Helminthological Society of Washington
284 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
next 8 years, she worked as a biologist in the Animal <strong>Parasitology</strong> Institute (API) of the U.S.<br />
Department of Agriculture doing research on poultry and bovine parasites. Shortly after her arrival<br />
at API, I received a call from her supervisor, Dr. Vetterling, saying that he had hired a new person<br />
for his electron microscopy lab, and I should come over to meet her. That began our long scientific<br />
and social association. In 1976, Nancy moved to the Naval Medical Research Institute as a research<br />
microbiologist. There, she studied parasite immunology in relation to the development of malaria<br />
vaccines and later worked on cytokine regulation in wound repair. In 1994, she somehow slipped<br />
out of parasitology, without seeking advice of the Helminthological Society of Washington, and<br />
worked in the Wound Repair Program at NMRI until her retirement in 1997.<br />
Nancy's resume lists numerous publications. They illustrate her research contributions in the<br />
ultrastructure of intracellular parasites, techniques for isolation of large numbers of malaria parasites,<br />
which is a prerequisite to vaccine development, development of an oral vaccine against Campylobacter<br />
infection, development of a method to study local inflammatory action, and the study of the<br />
effects of cytokines in preventing translocation of bacteria in hemorrhagic shock.<br />
A predominant factor in the Committee's decision was Nancy's excellent service to the Society,<br />
of which, undoubtably, most of you are aware. She has a record that is hard to beat. To the best<br />
of my knowledge, and with a little help from her curriculum vitae, Nancy has served in every<br />
office and has been on every committee of the Society, with the exception of Editor and the Editorial<br />
Committee. After holding the position of vice president in 1980, she moved up to president the<br />
next year. She must have done something right because she was re-elected president in 1991, a feat<br />
that, with one exception, has been unmatched in recent Society history. In 1999, she was elected<br />
to the position of corresponding secretary-treasurer, which she currently holds.<br />
In spite of the considerable time that she has donated to the Society, Nancy has offered her<br />
expertise in various roles in other societies such as the American Society of Parasitologists and the<br />
American Society of Tropical Medicine and Hygiene. Outside of the scientific area, she has been<br />
active in many church-related events with her husband Jim. There is one other activity that might<br />
be noted. As mentioned before, Nancy spent a number of years working with poultry coccidia. You<br />
all know what people in that field do—they search through chicken droppings and count the coccidial<br />
oocysts that they find. Well, Nancy must have developed a high degree of excellence in<br />
counting because she has steadily moved upward through the ranks in the H&R Block organization<br />
and is now a senior tax preparer.<br />
Nancy, I am very pleased to present to you, on the behalf of the Anniversary Awards Committee,<br />
the Helminthological Society of Washington's Anniversary Award for 2000.<br />
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 284-285<br />
MINUTES<br />
Harley G. Sheffield, Ph.D.<br />
November 15, 2000<br />
Six Hundred Seventy-First through the Six Hundred Seventy-Fifth<br />
Meeting of the Helminthological Society of Washington<br />
671st Meeting: George Washington University, meeting. Dr. Eckerlin introduced the speakers.<br />
Washington, DC, 12 October 2000. President Robert Gwadz gave an overview of the National<br />
Dennis Richardson conducted the business Institutes of Health (NIH)-funded malaria remeeting.<br />
Ralph Eckerlin welcomed members search in Mali. This NIH program funds elecand<br />
their guests and introduced the President, lives for students with interests in either basic<br />
who briefly summarized the Executive Council sciences or clinical aspects of malaria. Albert<br />
Copyright © 2011, The Helminthological Society of Washington
Nieto reviewed recent advances in the use of<br />
synthetic peptides for cystic hydatid disease serology.<br />
John Hawdon provided an account of his<br />
research on potential hookworm vaccine candidates.<br />
Finally, Dr. Eckerlin reviewed his studies<br />
of fleas from flying squirrels in Virginia. New<br />
and renewal members included Russell C. Van<br />
Horn (U.S.A.), Eric Panitz (U.S.A.), and Riccardo<br />
Fiorillo (U.S.A.).<br />
672nd Meeting: 94th Aero Squadron, <strong>College</strong><br />
Park, Maryland, 15 November 2000. The anniversary<br />
dinner meeting and program were presided<br />
over by President Dennis Richardson. The<br />
slate of officers for <strong>2001</strong> were elected and installed<br />
by the membership in attendance: Dennis<br />
J. Richardson, president; William E. Moser, vice<br />
president; W. Patrick Carney and Nancy D. Pacheco<br />
continue as reporting secretary and corresponding<br />
secretary-treasurer, respectively.<br />
Nancy Pacheco was presented the Anniversary<br />
Award by Harley Sheffield. John S. Mackiewicz<br />
and Graham C. Kearn were elected to Life<br />
Membership (accepted for him by Gene Hayunga)<br />
and Honorary Membership (accepted for<br />
him by Sherman Hendrix), respectively.<br />
673rd Meeting: Nematology Laboratory, Agricultural<br />
Research Service, USDA, Beltsville,<br />
Maryland, 17 January <strong>2001</strong>. President Dennis<br />
Richardson presided over the business meeting,<br />
and Vice President Lynn Carta presided over the<br />
scientific session. Peter Maser summarized his<br />
paper "<strong>Comparative</strong> Biochemistry of Parasitic<br />
and Free-Living Nematodes." William Wergin<br />
gave an overview of "Low Temperature Scanning<br />
Microscopy and its Application in Agriculture."<br />
Dr. Carta finished the session with "Integrating<br />
Nematode Morphology and Molecules<br />
in Plant-Parasitic and Free-Living Nematodes."<br />
Two new members were announced: Wellington<br />
MINUTES 285<br />
A. Oyibo (Nigeria) and Analfa Cristina Paola<br />
(Argentina).<br />
674th Meeting: Walter Reed Army Institute of<br />
Research/Naval Medical Research Center, Silver<br />
Spring, Maryland, 12 March <strong>2001</strong>. President<br />
Dennis Richardson presided over the business<br />
meeting and Eileen Franke-Villasante presided<br />
over the scientific session. Ed Rowton reviewed<br />
the "Status of the Recent Outbreak of Canine<br />
Leishmaniasis in the United <strong>State</strong>s." His paper<br />
was followed by David Fryauff's report on "A<br />
New Twist on an Old Drug: Recent DOD Studies<br />
of Primaquine for Malaria Prophylaxis."<br />
Kent Kester summarized "Advances in Pre-<br />
Erythrocytic Malaria Vaccine Development,"<br />
and Dr. Ling presented the final paper on "The<br />
Labor Involved in Malaria Field Studies in Indonesia."<br />
675th Meeting: Biology Department, Gettysburg<br />
<strong>College</strong>, Gettysburg, Pennsylvania, 5 May<br />
<strong>2001</strong>. President Richardson presided over the<br />
business meeting. Robin Overstreet, vice president<br />
of the American Society of Parasitologists<br />
(ASP), was welcomed and introduced to the<br />
members and guests by the president. Dr. Overstreet<br />
advised members that ASP has resources<br />
to provide travel grants for students who present<br />
papers at ASP meetings and to support speakers<br />
who are invited to present papers at meetings of<br />
affiliated societies. Sherman Hendrix presided<br />
over the scientific session. The first paper, presented<br />
by Dr. Overstreet, covered "Shrimp Parasites<br />
and Diseases," followed by Eric Hoberg's<br />
paper on the "Ancient Mariner—A History of<br />
Seabirds and Tapeworms on the Deep Blue<br />
Sea." Ann Barse summarized "New Host and<br />
Geographic Records of Monogenea of Billfishes."<br />
Sherman Hendrix presented the final paper<br />
on "Some Aspects of Biology of Bothitrema<br />
bothi (Platyhelminthes: Monogenea)."<br />
Copyright © 2011, The Helminthological Society of Washington
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, p. 286<br />
Abaigar, T., 134<br />
Abrams, A., 177<br />
Agarwal, N., 87<br />
Aguilar-Aguilar, R., 204, 272<br />
Aguirre-Macedo, M. L., 42, 76,<br />
190<br />
Al-Sady, R. S. S., 108<br />
Amin, O. M., 103, 108<br />
Arjona-Torres, G., 42, 190<br />
Baez-Vale, R., 196<br />
Balfour, C. D., 126, 259<br />
Bassat, S. E, 108<br />
Ben Hassine, O. K., 91<br />
Boeger, W. A., 66<br />
Bolek, M. G., 164<br />
Brailovsky, D., 196<br />
Brooks, D. R., 177<br />
Burge, A. N., 52<br />
Bursey, C. R., 21, 138<br />
Cabanas-Carranza, G., 196, 204<br />
Caira, J. N., 52<br />
Cano, M., 134<br />
Carreno, R. A., 177<br />
Caspeta-Mandujano, J. M., 196, 204<br />
Center, T. D., 242<br />
Coggins, J. R., 164, 269<br />
Cone, D. K., 236<br />
Cremonte, E, 277<br />
Criscione, C. D., 149, 156<br />
Curran, S. S., 219<br />
Curtis, M. A., 112<br />
Damborenea, M. C., 249<br />
Dang, T. T., 219<br />
Davies, K. A., 242<br />
Demarais, S., 116<br />
Diaz, J. I., 277<br />
Dumailo, S., 42, 190<br />
Durden, L. A., 177<br />
Eckerlin, R. P., 143<br />
Espeso, G., 134<br />
Comp. Parasitol.<br />
<strong>68</strong>(2), <strong>2001</strong>, pp. 286-291<br />
AUTHOR INDEX FOR VOLUME <strong>68</strong><br />
Euzet, L., 91<br />
Fedynich, A. M., 116<br />
Font, W. E, 149, 156<br />
Forrester, D. J., 130, 173<br />
Foster, G. W., 130, 173<br />
Fried, B., 126, 256, 259<br />
Garcia-Prieto, L., 9<br />
Garijo, M. M., 134<br />
Giblin-Davis, R. M., 242<br />
Goldberg, S. R., 21, 138<br />
Gonzalez-Sob's, D., 42, 190<br />
Hasegawa, H., 261<br />
Hoberg, E. P., 177<br />
Hogue, C. C., 36<br />
Huffman, J. E., 256<br />
Jimenez-Ruiz, E A., 9<br />
Joy, J. E., 185<br />
Kharchenko, V. A., 97<br />
Kinsella, J. M., 130, 274<br />
Krecek, R. C., 97<br />
Kritsky, D. C., 66, 87<br />
Kuperman, B. I., 274<br />
Kuzmin, Y., 228<br />
Lane, S. J., 274<br />
Lichtenfels, J. R., 97, 265<br />
Lloyd, G. E, 274<br />
Love, S., 265<br />
Matey, V. E., 274<br />
Matthews, J. B., 265<br />
Mayen-Pena, E., 196<br />
McDonnell, A., 265<br />
Mendoza-Garfias, B., 9<br />
Mhaisen, E T., 108<br />
Milek, J. A., 122<br />
Monasmith, T., 116<br />
Moreno-Navarrete, R. G., 204, 272<br />
Mullican, S. K., 256<br />
Navone, G. T., 277<br />
Neifar, L., 91<br />
Nguyen, T. L., 219<br />
Ortiz, J. M., 134<br />
Overstreet, R. M., 219<br />
Paola, A., 249<br />
Parmelee, J. R., 21<br />
Peng, J. S., 36<br />
Perez-Ponce de Leon, G., 1, 9<br />
Pham, X.-D., 261<br />
Posel, P., 42, 190<br />
Reddy, A., 259<br />
Reinbold, J. C., 269<br />
Robaldo, R. B., 66<br />
Rossi, M. L., 126<br />
Ruiz de Ybanez, M. R., 134<br />
Salgado-Maldonado, G., 196, 204,<br />
272<br />
Sanchez-Nava, P., 204<br />
Scholz, T., 42, 76, 190<br />
Seville, R. S., 122<br />
Siu-Estrada, E., 42, 190<br />
Snyder, S. D., 228<br />
Soto-Galera, E., 196, 204<br />
Thul, J. E., 173<br />
Tkach, V. V., 228<br />
Tran, C.-L., 261<br />
Tucker, R. B., 185<br />
Vidal-Martinez, V. M., 42, 76, 190<br />
Villa-Ramirez, B., 272<br />
Williams, D. S., 242<br />
Wright, M. E., 112<br />
Yadav, V. S., 87<br />
Yoder, H. R., 269<br />
KEYWORD AND SUBJECT INDEX FOR VOLUME <strong>68</strong><br />
Acanthobothrium dollyae sp. n., 52<br />
Acanthobothrium maryanskii sp.<br />
n., 52<br />
Acanthobothrium royi sp. n., 52<br />
Acanthocephala, 9, 36, 103, 108,<br />
130, 190, 196, 204, 261, 272<br />
286<br />
Copyright © 2011, The Helminthological Society of Washington<br />
Acanthocephalan cystacanth, 21<br />
Acuariidae gen. sp., 21, 196<br />
Adenomera andreae, 21
Adenomera hylaedactyla, 21<br />
Algansea tincella, 204<br />
Algonquin Park, Canada, 236<br />
Allocreadiidae gen. sp., 190<br />
Allocreadium mexicanum, 204<br />
Ancyrocephalus etropli, 87<br />
Ambystoma maculatum, 228<br />
American toad, 228<br />
Amphibia, 1, 21, 149, 164, 185,<br />
228, 269<br />
Amphilophus alfari, 42, 76, 190<br />
Anas platyrhynchus, 1<br />
Anisakis sp., 36<br />
Anisakis typica, 272<br />
Anolis carolinensis, 149<br />
Anura, 1, 21, 149, 164, 185<br />
Apharyngostrigea sp., 42<br />
Apicomplexa, 122<br />
Aplectana hylambatis, 21<br />
Aponurus sp., 36<br />
Archocentrm nigrofasciatus, 42,<br />
76<br />
Arctic chair, 112<br />
Arctic lakes, 112<br />
Argentina, 249, 277<br />
Ascocotyle (Ascocotyle) tenuicollis,<br />
42<br />
Ascocotyle (Phagicola) diminuta,<br />
42<br />
Ascocotyle (Phagicola) mollienisicola,<br />
42<br />
Ascocotyle (Phagicola) nana, 42<br />
Ashworthius sp., 177<br />
Aspidogastrea, 249<br />
Astyanax fasciatus, 1, 42, 190,<br />
196<br />
Athene cunicularia, 130<br />
Atherinella crystallina, 204<br />
Australia, 242<br />
Autonomous Region of the South<br />
Atlantic, Nicaragua, 42<br />
Aves, 1, 130, 173, 277<br />
Awaous tajasica, 204<br />
Aythya collaris, 173<br />
Azteca chub, 204<br />
Baja California Sur, Mexico, 272<br />
Balsas River, Mexico, 196<br />
Balsas catfish, 196<br />
Balsas shiner, 196<br />
Balsas splitfin, 196<br />
Barn owl, 130<br />
Barred owl, 130<br />
Basin white-lipped frog, 21<br />
Batracholandros magnavulvaris,<br />
185<br />
Batracholandros spectatus, 21<br />
Beireis' treefrog, 21<br />
Biodiversity, 1, 177<br />
Biogeography, 149<br />
Biomphalaria glabrata, 126<br />
Black-bellied garter snake, 9<br />
Blackbelt cichlid, 42, 76, 190<br />
Blackfin goodea, 196, 204<br />
Blackfin silverside, 204<br />
Blood parasites, 173<br />
Bluegill, 204<br />
Bolbosoma hamiltoni, 272<br />
Bolivian bleating frog, 21<br />
Bolivian white-lipped frog, 21<br />
Boophilus microplus, 177<br />
Bothriocephalus acheilognathi, 9,<br />
196, 204<br />
Brachylairna mcintoshi, 130<br />
Brachylecithum rarum, 130<br />
Brazil, 66<br />
Brevimulticaecum sp., 21, 42<br />
Brilliant-thighed poison frog, 21<br />
Broad-leaved paperbark, 242<br />
Brown egg frog, 21<br />
Brown newt, 228<br />
Bubo virginianus, 130<br />
Buenos Aires Province, Argentina,<br />
249<br />
Bufo americanus, 228<br />
Bufo glaberrimus, 21<br />
Bufo marinus, 1, 21<br />
Bufo typhonius, 21<br />
Burchell's zebra, 97<br />
Burrowing owl, 130<br />
Butterfly cichlid, 42, 76, 190<br />
California, U.S.A., 36, 274<br />
Canada, 112, 236<br />
Cane toad, 1, 21<br />
Capillaria cyprinodonticola, 196<br />
Capillaria dispar, 130<br />
Capillaria falconis, 130<br />
Capillaria sp., 130<br />
Capillaria tenuissima, 130<br />
Capillariidae gen. sp., 204<br />
Capillariinae gen. sp., 9<br />
Casmerodius albus, 1<br />
Catenotaenia sp., 116<br />
Centrocestus formosanus, 204<br />
Centropomidae, 66<br />
Centropomus undecimalis, 66<br />
Centrorhynchiis kuntzi, 130<br />
Centrorhynchus spinosus, 130<br />
Centrocestus formosanus, 196<br />
Cepedietta michiganensis, 185<br />
Cestoda, 21, 42, 52, 112, 116,<br />
130, 138, 149, 156, 164, 196,<br />
261, 269, 272<br />
Cestoidea, 9, 36<br />
Copyright © 2011, The Helminthological Society of Washington<br />
INDEX 287<br />
Chandleronema longigutturata,<br />
130<br />
Channel catfish, 196<br />
Chelonia mydas, 1<br />
Checklist, 204<br />
Chihuahuan Desert, Mexico, 116<br />
Chirostoma humboldtianiim, 204<br />
Chirostoma jordani, 204<br />
Chirostoma labarcae, 204<br />
Chirostoma riojai, 206<br />
Choanotaenia speotytonis, 130<br />
Chubut, Argentina, 277<br />
Cichlasoma beani, 204<br />
Cichlasoma geddesi, 1<br />
Cichlasoma faceturn, 249<br />
Cichlasoma istlanum, 196<br />
Cichlasoma maculicauda, 42, 76,<br />
190<br />
Cichlasoma managuense, 42, 76,<br />
190<br />
Cichlasoma nigrofasciatum, 196<br />
Cichlasoma synspillum, 1<br />
Cichlasoma urophthalmus, 1<br />
Ciliata, 185<br />
Cladocystis trifolium, 42<br />
Clethrionomys gapperi, 122<br />
Clinical effects, 256<br />
Clinostoinum complanatum, 42,<br />
196, 204<br />
Clinostoinum sp., 42, 269<br />
Coccidia, 134<br />
Coccidiosis, 134<br />
Colostethus marchesianus, 21<br />
Colubridae, 219<br />
Common carp, 204<br />
Common oval frog, 21<br />
Common snook, 66<br />
Community ecology, 116<br />
Component communities, 1 16<br />
Contracaecum sp., 9, 42, 196, 204<br />
Convict cichlid, 42, 76, 190, 196<br />
Copepods, 112<br />
Coronocyclus coronatus, 265<br />
Coronocyclus labiatus, 265<br />
Coryrnbia ptychocarpa, 242<br />
Coiynosoma sp., 36<br />
Cosmocephalus argentinensis, 277<br />
Cosmocephalus obvelatus, 277<br />
Cosmocerca brasiliense, 21<br />
Cosmocerca parva, 21<br />
Cosmocerca podicipinus, 21<br />
Cosmocercella phyllomedusae, 21<br />
Cosmocercoides sp., 164, 269<br />
Costa Rica, 177<br />
Cosymbotus platyurus, 138<br />
Cow Knob salamander, 185<br />
Crassicutus cichlasomae, 190
288 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
Craterostomum acuticaudatum,<br />
265<br />
Crepidostomum sp., 190<br />
Crowding effects, 156<br />
Crump's treefrog, 21<br />
Cryptogonimidae gen. sp., 42, 204<br />
Ctenophryne geayi, 21<br />
Cundinamarca toad, 21<br />
Cuticle, 242<br />
Cuvier's gazelle, 134<br />
Cyathocotylidae, 219<br />
Cyathostominea, 265<br />
Cyathostomurn catinatum, 265<br />
Cyathostomum montgomeryi, 97<br />
Cyathostomurn pateratum, 265<br />
Cylicocyclus ashworthi, 265<br />
Cylicocyclus elongatus, 265<br />
Cylicocyclus insigne, 265<br />
Cylicocyclus leptostomum, 265<br />
Cylicocyclus nassatus, 265<br />
Cylicocyclus radiatus, 265<br />
Cylicocyclus ultrajectinus, 265<br />
Cylicostephanus bidentatiis, 265<br />
Cylicostephanus calicatus, 265<br />
Cylicostephanus longibursatus,<br />
265<br />
Cylicostephanus minutus, 265<br />
Cylicostephanus goldi, 265<br />
Cylidodontophorus bicoronatus,<br />
265<br />
Cylindrotaenia americana, 21<br />
Cynops ensicauda, 228<br />
Cyprinus carpio, 204<br />
Cyst storage, 126<br />
Cysticercus, 116, 177<br />
Dactylogyridae, 76, 87<br />
Darkedged splitfin, 204<br />
Demerara Falls treefrog, 21<br />
Dermatemys mawii, 1<br />
Development, 149<br />
Diaptomus (Aglaodiaptornus) leptopus,<br />
112<br />
Diaptomus (Leptodiaptornus) minutus,<br />
112<br />
Didelphis virginiana, 1, 274<br />
Didelphostrongylus hayesi, 274<br />
Digenea, 1, 9, 36, 42, 190, 196,<br />
204<br />
Diphyllobothrium dendriticum,<br />
112<br />
Diplectanidae, 66<br />
Diplobatis ommata, 52<br />
Diplostomum sp., 204<br />
Diplostomum (A.) compactutn, 42,<br />
196<br />
Diplostomum (Tylodelphys) sp., 9<br />
Dipodomys merriami, 116<br />
Dispharynx affinis, 130<br />
Dispharynx nasuta, 130<br />
Domestic chicks, 256<br />
Dorcas gazelle, 134<br />
Dracunculus ophidensis, 9<br />
Dull rocket frog, 21<br />
Eastern screech-owl, 130<br />
Echinostoma caproni, 126, 256,<br />
259<br />
Echinostoma revolutum, 256<br />
Echinostoma trivolvis, 256<br />
Echinostomiasis, 256<br />
Ecology, 261<br />
Edalorhina perezi, 21<br />
Eimeria clethrionomyis, 122<br />
Eimeria elegatis, 134<br />
Eimeria gazella, 134<br />
Eimeria pallida, 134<br />
Eirunepe snouted treefrog, 21<br />
Elachistocleis avails, 21<br />
Elasmobranchii, 52, 91<br />
Electron microscopy, 52, 228,<br />
242, 249, 274, 277<br />
Eleutherodactylus cruralis, 21<br />
Eleutherodactylus fenestratus, 21<br />
Eleutherodactylus imitatrix, 21<br />
Eleutherodactylus peruvianus, 21<br />
Eleutherodactylus toftae, 21<br />
Ency station, 126<br />
Enhydris chinensis, 219<br />
Enhydris plumbea, 219<br />
Epidermis, 242<br />
Epipedobates femoralis, 21<br />
Epipedobates pictus, 21<br />
Equus burchelli antiquorum, 97<br />
Zssctt: americanus, 103<br />
Estado de Mexico, Mexico, 9,<br />
196, 204<br />
Etroplus suratensis, 87<br />
Eucalyptus leucoxylon, 242<br />
Eustrongylides sp., 9, 42, 196,<br />
204<br />
Excisa excisiformis, 130<br />
Excretion pattern, 134<br />
Excystation, 126, 259<br />
Experimental infection, 112<br />
Falcaustra affinis, 9<br />
Falcaustra mexicana, 9<br />
Falcaustra wardi, 9<br />
Falcaustra sp., 42<br />
Fence lizards, 149<br />
Fergusobia sp., 242<br />
Fergusonina sp., 242<br />
Filaria sp., 116<br />
Fixation artifacts, 156, 219<br />
Flat-bodied house gecko, 138<br />
Copyright © 2011, The Helminthological Society of Washington<br />
Florida, U.S.A., 130, 149, 173<br />
Fourth International Congress of<br />
Nematology, 172<br />
Fourth International Symposium<br />
on Monogenea, 142<br />
Freshwater mullet, 108<br />
Gazella cuvieri, 134<br />
Gazella dama mhorr, 134<br />
Gazella dorcas neglecta, 134<br />
Gazelle, 134<br />
Gehyra mutilata, 138<br />
Gekkonidae, 138, 149<br />
Genyonemus lineatus, 36<br />
Georgia, U.S.A., 103<br />
Girardinichthys multiradiatus, 204<br />
Glossocercus auritus, 196<br />
Glypthelmins parva, 21<br />
Glypthelmins quieta, 164, 269<br />
Gnathostoma sp., 204<br />
Goblet cell, 256<br />
Gold-striped frog, 21<br />
Golden hamster, 112<br />
Gongylonema dipodomysis, 116<br />
Gongylonema neoplasticum, 261<br />
Gongylonema pulchrum, 177<br />
Goodea atripinnis, 196, 204<br />
Gorgoderina bilobata, 269<br />
Great horned owl, 130<br />
Green anole, 149<br />
Green chromide, 87<br />
Green frog, 164, 269<br />
Guanajuato, Mexico, 204<br />
Guerrero, Mexico, 196<br />
Gulf of California, 52<br />
Gunther's banded treefrog, 21<br />
Guppy, 196<br />
Gussevia herotilapiae sp. n., 76<br />
Gyalocephalus capitatus, 265<br />
Gymnura altavela, 91<br />
Gyrodactylus elegans, 204<br />
Gyrodactylus sp., 196, 204<br />
Hadwenius tursionis, 212<br />
Haemaphysalis juxtakochi, 177<br />
Haematoloechus varioplexus, 164,<br />
269<br />
Haemoproteus nettionis, 173<br />
Halipegus eccentricus, 164, 269<br />
Hamptophryne boliviano, 21<br />
Hedruris siredonis, 9<br />
Heleobia parchappii, 249<br />
Helminth larvae, 42<br />
Helminthological Society of<br />
Washington:<br />
Anniversary Award, 283<br />
Dues Increase, 163<br />
Editors' Acknowledgments, 155
Honorary Membership, 282<br />
Life Membership, 282<br />
Meeting Schedule, 121<br />
Mission and Vision <strong>State</strong>ments,<br />
147<br />
Symposium Announcement,<br />
227<br />
Helminths, 9, 36, 116, 130, 149,<br />
156, 177, 185, 190, 196, 204,<br />
219, 228, 242, 249, 256, 259,<br />
261, 265, 269, 272, 274, 277<br />
Hematozoa, 173<br />
Hemidactylus frenatus, 138<br />
Hemidactylus garnotii, 138, 149<br />
Hemidactylus turcicus, 149, 156<br />
Hemiphyllodactylus typus, 138<br />
Henle's snouted treefrog, 21<br />
Herotilapia multispinosa, 42, 76,<br />
190<br />
Heterandria bimaculatus, 196<br />
Heteromoxyuris deserti, 116<br />
Heteronchocotyle gymnurae sp.<br />
n., 91<br />
Heterophyidae gen. sp., 42<br />
Heterostrongylus heterostrongylus,<br />
274<br />
Hexabothriidae, 91<br />
Horses, 265<br />
Host specificity, 149<br />
House gecko, 138<br />
Hybopis boucardi, 196<br />
Hyla boans, 21<br />
Hyla brevifrons, 21<br />
Hyla calcarata, 21<br />
Hyla fasciata, 21<br />
Hyla granosa, 21<br />
Hyla koechlini, 21<br />
Hyla leali, 21<br />
Hyla leucophyllata, 21<br />
Hyla marmorata, 21<br />
Hyla parviceps, 21<br />
Hyla rhodopepla, 21<br />
Hyla schubarti, 21<br />
Hymenolepis diminuta, 261<br />
Hysterothylacium sp., 196<br />
Ictalunis balsanus, 196<br />
Ictalurus punctatus, 196<br />
Ilyodon whitei, 196<br />
Imitator robber frog, 21<br />
India, 87<br />
Indo-Pacific geckos, 138, 149<br />
Infectivity, 126, 256<br />
Insecta, 242<br />
Iraq, 108<br />
Ixodes affinis, 177<br />
Jaguar cichlid, 42, 76, 190<br />
Jaguar leaf frog, 21<br />
Jalisco, Mexico, 9, 196, 204<br />
Japan, 228<br />
Jeweled splitfin, 204<br />
Juvenile nematodes, 36<br />
Key, 236<br />
Kinosternon hirtipes, 9<br />
Koechlin's treefrog, 21<br />
Lacistorhynchus dollfusi, 21<br />
La Paz robber frog, 21<br />
Leal's treefrog, 21<br />
Lecithochirium sp., 21<br />
Lemdana wernaati, 130<br />
Lepidochelys olivacea, 1<br />
Lcpidodactylus aurcolincatus, 138<br />
Lepornis gibbosus, 236<br />
Lepomis macrochirus, 206<br />
Leptodactylus bolivianus, 21<br />
Leptodactylus leptodactyloides, 21<br />
Leptodactylus mystaceus, 21<br />
Leptodactylus pentadactylus, 21<br />
Leptodactylus petersii, 21<br />
Leptodactylus rhodonotus, 21<br />
Lerma chub, 204<br />
Lerma livebearer, 196, 204<br />
Lerma-Santiago river basin, Mexico,<br />
204<br />
Leucocytozoon simondi, 173<br />
Life cycle, 112<br />
Ligula intestinalis, 204<br />
Lithodytes lineatiis, 21<br />
Liza abu, 108<br />
Lobatostoma jungwirthi, 249<br />
Long-whiskered catfish, 190<br />
Los Angeles Harbor, U.S.A., 36<br />
Louisiana, U.S.A., 149, 156<br />
Lowland tropical bullfrog, 21<br />
Madre de Dios treefrog, 21<br />
Magellanic penguin, 277<br />
Magnivitellinum simplex, 190, 196<br />
Mammalia, 1, 97, 112, 116, 122,<br />
126, 134, 177, 259, 261, 265,<br />
272, 274<br />
Manaus slender-legged treefrog,<br />
21<br />
Manteriella sp., 36<br />
Marbled treefrog, 21<br />
Margotrema bravoae, 204<br />
Maritrema sp., 130<br />
Mastophorus dipodomis, 116<br />
Mediterranean geckos, 149, 156<br />
Mediterranean Sea, 91<br />
Megalodiscus temperatus, 269<br />
Melaleuca quinquenervia, 242<br />
Merriam's kangaroo rat, 116<br />
Copyright © 2011, The Helminthological Society of Washington<br />
INDEX 289<br />
Mesa Central, Mexico, 9<br />
Mesa silverside, 204<br />
Mesocestoides sp., 164, 269<br />
Mesocricetus auratus, 112<br />
Metacercariae, 42, 126, 164, 259,<br />
269<br />
Mexican garter snake, 9<br />
Mexican molly, 196, 204<br />
Mexican mud turtle, 9<br />
Mexican tetra, 196<br />
Mexico, 1, 9, 52, 196, 204, 272<br />
Michoacan, Mexico, 9, 196, 204<br />
Microfilaria, 173<br />
Microphallus sp., 130<br />
Mississippi, U.S.A., 149<br />
Mohor gazelle, 134<br />
Mollusca, 126, 249<br />
Molly, 190<br />
Molting, 242<br />
Moniliformis moniliformis, 261<br />
Monogenea, 9, 76, 91, 196, 204<br />
Monogenoidea, 66, 87<br />
Morelos, Mexico, 196<br />
Morphology, 97, 156, 265, 277<br />
Mugilidae, 108<br />
Mus musculus, 126, 259<br />
Myrtaceae, 242<br />
Myxobolus cyprinicola, 236<br />
Myxobolus dechtiari, 236<br />
Myxobolus gibbosus, 236<br />
Myxobolus Hi, 236<br />
Myxobolus lipomicus, 236<br />
Myxobolus magnaspherus, 236<br />
Myxobolus osburni, 236<br />
Myxobolus paralintoni, 236<br />
Myxobolus poecilichthidis, 236<br />
Myxobolus uvuliferus, 236<br />
Myxozoa, 236<br />
Namibia, 97<br />
Napo's tropical bullfrog, 21<br />
Natalus mexicanus, 1<br />
Nayarit, Mexico, 204<br />
Nematoda, 9, 21, 42, 97, 116,<br />
130, 138, 164, 185, 190, 196,<br />
204, 228, 242, 261, 265, 269,<br />
272, 274, 277<br />
Neodiplostomum americanum, 130<br />
Neodiplostomum reflexum, 130<br />
Neoechinorhynchidae, 103, 108<br />
Neoechinorhynchus didelphis sp.<br />
n., 103<br />
Neoechinorhynchus golvani, 190,<br />
196, 204<br />
Neoechinorhynchus iraqensis sp.<br />
n., 108<br />
New books, 102, 107, 176, 255<br />
New combination, 87, 219
290 COMPARATIVE PARASITOLOGY, <strong>68</strong>(2), JULY <strong>2001</strong><br />
New genus, 87<br />
New geographical record, 9, 21,<br />
42, 52, 66, 76, 87, 122, 130,<br />
134, 138, 173, 177, 185, 190,<br />
196, 204, 236, 261, 265, 269,<br />
272, 274, 277<br />
New host record, 9, 21, 36, 42,<br />
52, 66, 76, 87, 112, 122, 130,<br />
134, 138, 164, 177, 185, 204,<br />
261, 272, 274, 277<br />
New Mexico, U.S.A., 116<br />
New species, 52, 76, 91, 103,<br />
108, 219, 228<br />
New synonymy, 236<br />
Nicaragua, 42, 76, 190<br />
Nippostrongylus brasiliensis, 261<br />
Northern leopard frog, 228<br />
Notocotylus sp., 261<br />
Notropis sallei, 204<br />
Nycticorax nycticorax, 1<br />
Oaxaca, Mexico, 196<br />
Obituary:<br />
W. Henry Leigh, 143<br />
Obituary Notice:<br />
Julius Feldmesser, 41<br />
Ocellated electric ray, 52<br />
Ochetosoma brevicaecum, 9<br />
Ochoterenella vellardi, 21<br />
Odocoileus virginianus, 177<br />
Oligogonotylus manteri, 42, 190<br />
Ommatobrephidae, 219<br />
Ondoorstepoort Helminthological<br />
Collection, 195<br />
Ontario, Canada, 236<br />
Oochoristica javaensis, 138, 149,<br />
156<br />
Oocyst, 122, 134<br />
Ophidascaris sp., 21<br />
Orientostrongylus cf. tenorai, 261<br />
Orinoco lime treefrog, 21<br />
Osteocephalus taurinus, 21<br />
Oswaldocruzia lopesi, 21<br />
Oswaldocruzia pipiens, 164, 269<br />
Otus asio, 130<br />
Pachitea robber frog, 21<br />
Parapharyngodon maplestoni, 138<br />
Parelaphostrongylus tennis, 177<br />
Parvitaenia cochlearii, 196<br />
Parvitaenia macropeos, 196<br />
Pastel cichlid, 42, 76, 190<br />
Pathology, 256<br />
Perciformes, 66, 87<br />
Perez's snouted frog, 21<br />
Pernambuco, Brazil, 66<br />
<strong>Peru</strong>, 21<br />
<strong>Peru</strong> robber frog, 21<br />
<strong>Peru</strong> white-lipped frog, 21<br />
Petenia splendida, 1<br />
Peter's frog, 21<br />
Phalacrocorax olivaceus, 1<br />
Philander opossum, 1<br />
Philippine Islands, 138<br />
Phrynohyas coriacea, 21<br />
Phrynohyas venulosa, 21<br />
Phyllomedusa atelopoides, 21<br />
Phyllomedusa pcdliata, 21<br />
Phyllomedusa tomopterna, 21<br />
Phyllomedusa vaillanti, 21<br />
Physaloptera sp., 21<br />
Physalopteroides venancioi, 21<br />
Pipa pipa, 21<br />
Pisces, 1, 36, 42, 66, 76, 87, 103,<br />
108, 112, 190, 196, 204, 236,<br />
249<br />
Plagiorchis sp., 130<br />
Platyhelminthes, 1<br />
Plethodon punctatus, 185<br />
Plethodon wehrlei, 185<br />
Pneumatophilus variabilis, 9<br />
Poecilia reticulata, 196<br />
Poecilia sphenops, 196, 204<br />
Poecilia veil/era, 42, 190<br />
Poeciliopsis gracilis, 196<br />
Poeciliopsis infans, 196, 204<br />
Poeciliopsis sp., 204<br />
Polymorphic brevis, 9, 204<br />
Polystornoidella oblonga, 9<br />
Porrocaecum depressmn, 130<br />
Porrocaecum sp., 21<br />
Porthole livebearer, 196<br />
Posthodiplostomurn minimum, 9,<br />
42, 196, 204<br />
Postorchigenes ovatus, 138<br />
Prescribed fire, 116<br />
Procamallanus (Spirocamallanus)<br />
neocaballeroi, 190<br />
Procamallanus (Spirocamallanus)<br />
rebecae, 190<br />
Prosthenhystera obesa, 190<br />
Prosthogonirnus ovatus, 130<br />
Proteocephalidea gen. sp., 42, 204<br />
Proteocephalus sp., 269<br />
Proteocephalus variabilis, 9<br />
Proterodiplostomidae gen. sp., 42<br />
Proterodiplostomum sp., 204<br />
Pseudis paradoxa, 21<br />
Pseudocapillaria tomentosa, 204<br />
Pterygodermatites dipodomis, 116<br />
Puebla, Mexico, 196<br />
Puerto Rico, 66<br />
Pumpkinseed sunfish, 236<br />
Quebec, Canada, 112<br />
Queretero, Mexico, 204<br />
Copyright © 2011, The Helminthological Society of Washington<br />
Queensland, Australia, 242<br />
Raillietina celebensis, 261<br />
Raillietnema gubernaculatum, 21<br />
Rana clamitans, 269<br />
Rana clamitans melanota, 164<br />
Rana montezumae, 1<br />
Rana pipiens, 228<br />
Rana sphenocephala, 149<br />
Rana vaillanti, 1<br />
Ransom Trust Report, 248<br />
Rattus argiventer, 261<br />
Rattus losea, 261<br />
Rattus tanezumi, 261<br />
Rear-fanged water snake, 219<br />
Redescription, 236<br />
Redfin pickerel, 103<br />
Redside cichlid, 196<br />
Red-skirted treefrog, 21<br />
Red-snouted treefrog, 21<br />
Reptilia, 1, 9, 138, 149, 156, 219<br />
Reserva Cuzco Amazonico, 21<br />
Rhabdias ambystomae sp. n., 228<br />
Rhabdias americanus, 228<br />
Rhabdias bermani, 228<br />
Rhabdias fuscovenosa, 9<br />
Rhabdias ranae, 228<br />
Rhabdias tokyoensis, 228<br />
Rhabdiasidae, 228<br />
Rhabdochona canadensis, 196<br />
Rhabdochona kidderi kidderi, 190,<br />
196<br />
Rhabdochona lichtenfelsi, 196<br />
Rhabdochoma mexicana, 196<br />
Rhabdosynochus hargisi sp. n.,<br />
66<br />
Rhabdosynochus hudsoni sp. n.,<br />
66<br />
Rhabdosynochus rhabdosynochus,<br />
66<br />
Rhamdia nicaraguensis, 42, 190<br />
Ring-necked duck, 173<br />
Rio Mamore robber frog, 21<br />
River goby, 204<br />
Russia, 228<br />
Rusty treefrog, 21<br />
Saccocoelioides sogandaresi, 190,<br />
196<br />
Saccocoelioides spp., 190<br />
Salamanders, 185, 228<br />
Salamandrella kaiserlingii, 228<br />
Salmonidae, 112<br />
Salvelinus alpinus, 112<br />
Sarayacu treefrog, 21<br />
Scarthyla ostinodactyla, 21<br />
Sceloporus u. iindulatus, 149<br />
Schrankiana inconspicata, 21
Schrankiana larvata, 21<br />
Schrankiana schrankai, 21<br />
Schrankianella brasili, 21<br />
Schubart's Rondonia treefrog, 21<br />
Sciadicleithrum bicuensc sp. n.,<br />
76<br />
Sciadicleithrum maculicaudae sp.<br />
n., 76<br />
Sciadicleithrum meekii, 76<br />
Sciadicleithrum mexicanum, 76<br />
Sciadicleithrum nicaraguense sp.<br />
n., 76<br />
Sciadicleithrum sp., 204<br />
Scinax garbei, 21<br />
Scinax icterica, 21<br />
Scinax pedromedinai, 21<br />
Scinax ruba, 21<br />
Sclerocleidoid.es gen. n., 87<br />
Sclerocleidoides etropli comb, n.,<br />
87<br />
Scotland, 265<br />
Seasonal study, 164<br />
Sensory papillae, 249<br />
Serpinema trispinosum, 9, 42<br />
Sharpnose silverside, 204<br />
Shortfin silverside, 204<br />
Siberian newt, 228<br />
Sinaloan cichlid, 204<br />
Singhiatrema vietnamensis sp. n.,<br />
219<br />
Skrjabinelazia machidai, 138<br />
South Africa, 97<br />
South American bullfrog, 21<br />
South American common toad, 21<br />
South Australian blue gum, 242<br />
Southern leopard frog, 149<br />
Southern red-backed vole, 122<br />
Spain, 134<br />
Spauligodon hemidactylus, 138<br />
Sphaenorhynchus lacteus, 21<br />
Spheniscus magellanicus, 211<br />
Spinitectus osorioi, 9<br />
Spinner dolphin, 272<br />
Spiny butterfly ray, 91<br />
Spirocamallanus sp., 36<br />
Spiroxys contorta, 9<br />
Spiroxys sp., 42, 196, 204<br />
Spiroxys susanae, 9<br />
Spirurids, 130<br />
Splendidofilaria fallisensis, 173<br />
Spot-legged poison frog, 21<br />
Spottail chub, 204<br />
Spottail killifish, 196<br />
Spotted salamander, 228<br />
Stenella longirostris, 272<br />
Stephana stomum californicum, 36<br />
Stephanostomum tristephanum, 36<br />
Stomylotrema vicarium, 130<br />
Strigea elegans, 130<br />
Strix varia, 130<br />
Strobilocephalus triangularis, 272<br />
Strongylidae, 97<br />
Strongyloides ratti, 261<br />
Strongyloides sp., 130<br />
Strongyloides venezuelensis, 261<br />
.Stump-toed gecko, 138<br />
Stunkardiella minima, 42<br />
Subulura forcipata, 130<br />
Subulura reclinata, 130<br />
Surinam golden-eyed treefrog, 21<br />
Surinam toad, 21<br />
Survey, 9, 21, 42, 76, 130, 138,<br />
164, 173, 177, 185, 190, 196,<br />
261, 265, 269<br />
vSwamp bloodwood, 242<br />
Swimming frog, 21<br />
Synhimantus laticeps, 130<br />
Syphacia muris, 261<br />
Szidatia taiwanensis comb, n.,<br />
219<br />
Tadarida brasiliensis, 1<br />
Taxonomy, 9, 52, 66, 76, 87, 91,<br />
97, 103, 108, 149, 156, 204,<br />
219, 228, 236<br />
Tegument, 249<br />
Telorchis corti, 9<br />
Tetra, 42, 190<br />
Tetrameres microspinosa, 130<br />
Tetrameres strigiphila, 130<br />
Tetraphyllidea, 36, 52, 272<br />
Thailand, 138<br />
Thamnophis eques, 9<br />
Thamnophis melanogaster, 9<br />
Ticks, 177<br />
Tiger-striped leaf frog, 21<br />
Copyright © 2011, The Helminthological Society of Washington<br />
INDEX 291<br />
Toady leaf frog, 21<br />
Toluca silverside, 204<br />
Tree gecko, 138<br />
Trematoda, 21, 126, 130, 138,<br />
164, 249, 256, 259, 261, 269,<br />
272<br />
Tribolium castaneum, 149<br />
Tridentoinfundibulum gobi, 265<br />
Trigonocotyle sp., 272<br />
Troschel's treefrog, 21<br />
Tunisia, 91<br />
Tyto alba, 130<br />
Ultrastructure, 242, 249<br />
Urocledoides cf. costaricensis,<br />
196<br />
Uvulifersp., 42, 196<br />
U.S.A., 36, 103, 116, 122, 132,<br />
149, 156, 164, 173, 185, 228,<br />
269, 274<br />
Valipora minuta, 196<br />
Variation, 156<br />
Veined treefrog, 21<br />
Vietnam, 219, 261<br />
Virginia opossum, 274<br />
Waltonella sp., 164<br />
Wehrle's salamander, 185<br />
West Virginia, U.S.A., 185<br />
White croaker, 36<br />
White-lined leaf frog, 21<br />
White-tailed deer, 177<br />
Wisconsin, U.S.A., 164, 228, 269<br />
Worm recovery, 126<br />
Wyoming, U.S.A., 122<br />
Xenotoca variatus, 204<br />
Yellow-lined smooth-scaled gecko,<br />
138<br />
Yellow-snouted treefrog, 21<br />
Yucatan molly, 42<br />
Yurirea alta, 204<br />
Zalophotrema pacificum, 212<br />
Zebra, 97<br />
Zoogonoides sp., 36
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VOLUME <strong>68</strong> JULY <strong>2001</strong> NUMBER 2<br />
CONTENTS<br />
(Continued from Front Cover)<br />
CONE, D. K. Supplemental Diagnosis ofMyxobolus gibbosus (Myxozoa), with a Taxonomic Review of<br />
Myxobolids from Lepomis gibbosus (Centrarchidae) in North America . 236<br />
GIBLIN-DAVIS, R. M., K. A. DAVIES, D. S. WILLIAMS, AND T. D. CENTER. Cuticular Changes in Fergusobiid<br />
Nematodes Associated with Parasitism of Fergusoninid Flies 242<br />
PAOLA, A., AND M. C. OAMBORENEA. Tegumentary infrastructure (SEM) of Preadult and Adult Lobatostoma<br />
jungwiithi Kritscher, 1974 (Trematoda: Aspidogastrea) _ 249<br />
RESEARCH NOTES<br />
MULLIGAN, S. K., J. E. HUFFMAN, AND B. FRIED. Infectivity and <strong>Comparative</strong> Pathology of Echinostoma<br />
caproni, Echinostoma revolution, and Echinostoma trivolvis (Trematoda) in the Domestic Chick 256<br />
FRIED, B., A. REDDY, AND C. D. BALFOUR. Excystation and Distribution of Metacercariae of Echinostoma<br />
caproni in ICR Mice ... 259<br />
PHAM, X.-D., C.-L. TRAN, AND H. HASEGAWA. Helminths Collected from Rattus spp. in Bac Ninh<br />
Province, Vietnam . 261<br />
LICHTENFELS, J. R., A. MCDONNELL, S. LOVE, AND J. B. MATTHEWS. Nematodes of the Tribe Cyathostominea<br />
(Strongylidae) Collected from Horses in Scotland 265<br />
YODER, H. R., J. R. COGGINS, AND J. C. RjEiNBOLD. Helminth Parasites of the Green Frog (Rana clamitans)<br />
from Southeastern Wisconsin, U.S.A 269<br />
AGUILAR-AGUILAR, R., R. G. MORENO-NAVARRETE, G. SALGADO-MALDONADO, AND B. VILLA-RAM!REZ.<br />
Gastrointestinal Helminths of Spinner Dolphins Stenella longirostris (Gray, 1828) (Cetacea: Delphinidae)<br />
Stranded in La Paz Bay, Baja California Sur, Mexico . 272<br />
MATEY, V E., B. I. KUPERMAN, J. M. KINSELLA, G. F. LLOYD, AND S. J. LANE. The Lung Nematodes<br />
(Metastrongyloidea) of the Virginia Opossum Didelphis virginiana in Southern California, U.S.A. 274<br />
DIAZ, J. I., G. T. NAVONE, AND F. CREMONTE. New Host and Distribution Records of Cosmocephahis<br />
obvelatus (Creplin, 1925) (Nematoda: Acuariidae), with Morphometric Comparisons 277<br />
ANNOUNCEMENTS<br />
EDITORS ' ACKNOWLEDGMENTS 15 5<br />
NOTICE OF DUES INCREASE 163<br />
FOURTH INTERNATIONAL CONGRESS OF NEMATOLOGY . . 172<br />
NEW 'BOOKS AVAILABLE 176,255<br />
ONDERSTEPOORT HELMINTHOLOGICAL COLLECTION . 195<br />
SYMPOSIUM: PARASITOLOGY IN SCIENCE AND SOCIETY , 227<br />
HELMINTHOLOGICAL SOCIETY OF WASHINGTON MEETING SCHEDULE 241<br />
REPORT ON THE BRAYTON H. RANSOM MEMORIAL TRUST FUND , 248<br />
LIFE MEMBERSHIP: JOHN S. MACKIEWICZ . ...... 282<br />
HONORARY MEMBERSHIP: GRAHAM C. KEARNE 282<br />
PRESENTATION OF 2000 ANNIVERSARY AWARD 283<br />
MINUTES OF THE MEETINGS OF THE HELMINTHOLOGICAL SOCIETY OF WASHINGTON 284<br />
AUTHOR INDEX FOR VOLUME <strong>68</strong> 286<br />
KEY WORD AND SUBJECT INDEX FOR VOLUME <strong>68</strong> 286<br />
MEMBERSHIP APPLICATION OF THE HELMINTHOLOGICAL SOCIETY OF WASHINGTON 292<br />
Date of publication, 31 July <strong>2001</strong><br />
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