This dissertation has been microfilmed exactly as received 68-12,857
McGRAW, James Carmichael, 1928- A STUDY OF COSMOCERCOIDES DUKAE (HOLL, 1928) WILKIE, 1930 (NEMATODA: COSMO- CERCIDAE) FROM AMPHIBIANS IN OHIO.
The Ohio State University, Ph.D., 1968 Zoology
University Microfilms, Inc., Ann Arbor, Michigan A STUDY OP COSMOCERCOIDES DUKAE (HOLL, 1928) WILKIE, 1930
(NEMATODA: COSMOCERCIDAE) PROM AMPHIBIANS IN OHIO
DISSERTATION
Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University
James Carmichael McGraw, A.B., M.S.
******
The Ohio S tate U niversity 1968
Approved by
// /Adviser/Adviser D§partm£ntnt of of Zoology Zoology and Entomology ACKNOWLEDGMENTS
The writer wishes to thank Dr. John L. Crites of the Department of Zoology and Entomology at Ohio State
University for furnishing nematodes from Ohio amphibians.
Thanks are also due the following persons who helped to locate nematode specimens: Dr. Barry Valentine, Department of Zoology and Entomology at Ohio State University; Dr.
Russell R. Williams, Waynesburg College; Dr. Paul D.
Harwood, Hess and Clark; Dr. W. G. Inglis, Brisith Museum;
W. W. Becklund, M.B. Chitwood and J.M. Humphrey, U.S.D.A.-
Beltsville Parasitology Laboratory; Dr. Roy C. Anderson,
University of Guelph; Dr. Robert Ph. Dollfus, Museum
National d'Histoire Naturelle, Paris; Dr. J. E. Alicata,
University of Hawaii-; V. N. Converse, University of
Adelaide; M. Bravo Hollis, National University of Mexico;
N. T. Talbot, Departmentcf Agriculture of the Territory of
Papua and New Guinea; W. A. Reid of the University of
Kansas; Dr. R. U. Gooding, University of Singapore; and the late A. C. Walton of Knox College.
Thanks are due Celeste Taft for identification of terrestrial molluscs; Dr. C. J. Zenisek and Warren Dolfin for aid in the identification of amphibians; Mr. Igors
Gortinskis and M. A. Ewert for translation of Russian;
i i Dean Harold J. Perkins of the Faculty of Science and
Mathematics at Plattsburgh State University College of
Arts and Science for aid and encouragement; Dr. Russell
Skavaril for statistical analyses; and Dr. Robert J.
Shields of City College, City University of New York, for many suggestions.
I would like to express my appreciation to Mrs.
Dorothy Schreck and the library staff for easing the literature search.
Special thanks go to Dr. Joseph N. Miller whose advice and criticisms throughout this study were very much appreciated.
iii VITA
March 20, 1928 Born - Martins Perry, Ohio
1951 ...... A.B., Oberlin College, Oberlin, Ohio
1955-1964 . . Graduate Assistant, Department of Zoology and Entomology, The Ohio S ta te U n iv ersity , Columbus, Ohio
1957 ...... M.S., The Ohio State University, Columbus, Ohio
1964-1968 . . Assistant Professor of Biology, State University College of Arts and Science, Plattsburgh, New York
FIELDS OP STUDY
Major Field: Zoology
Studies in Invertebrate Zoology. W. J. Kostir
Studies in Medical Entomology. C. E. Venard
Studies in Vertebrate Zoology. J. W. Price
Studies in Parasitology. J. N. Miller
i v CONTENTS
Page
ACKNOWLEDGMENTS ...... l i
VITA ...... iv
TABLES...... v ii
INTRODUCTION ...... 1
HISTORICAL REVIEW ...... 4
MATERIALS AND METHODS ...... 11
OBSERVATIONS AND RESULTS ...... 15
Occurrence of Cosmocercoid.es dukae ...... 15 Morphology of Cosmocercoides' dukae ...... 20 C u t i c l e ...... 21 Cephalic region ...... 23 E s o p h a g u s...... 24 I n t e s t i n e ...... 25 Female reproductive system ...... 26 Male reproductive system ...... 28 Special male anatomy ...... 29 Descriptive Summary of Cosmocercoldes dukae from Ohio amphibians ...... 30 Observations on the Holotype of Cosmocercoldes dukae (Holl, 1928) ...... 33 Comparison of Selected Groups of Cosmocercoldes dukae (Holl, 1928) ...... 34 Observations on Cosmocercoldes pulcher W ilkie, 1930 ...... 52 Additions to the Description of Cosmocercoldes p u lch er—W ilk ie, 1930 54 An Emended D escription of the Genus Cosmocercoides—W ilkie, 1930 ...... 57 Observations on the Life History of Cosmocercoides dukae (Holl, 1928 ...... 58
DISCUSSION...... 68
SUMMARY......
v CONTENTS (Contd.)
Page
APPENDIX...... 82
LITERATURE CITED...... 87
PLATES...... 91
v i TABLES
T able Page
1. Cosmocercoldes dukae from Ohio Amphibians, A pril 19 6 3-July 1964 16
2. Measurements (in mm) of Cosmocercoides dukae from one American toad (Bufo amerlcanus) from Ohio and C. dukae reported by Anderson (I960) from terrestrial molluscs in Ontario . . 36
3. Derived statistics (percentages) ofC. dukae from one American toad from Ohio and C. dukae reported by Anderson (i 960 ) from terrestrial molluscs in Ontario ...... 37
4. Male measurements, Group I ...... 39
5. Female measurements, Group I ...... 40
6 . Male derived s t a t i s t i c s , Group I ...... 41
7. Female derived statistics, Group I ...... 42
8. Male measurements, Group I I ...... 43
9. Female measurements, Group I I ...... 44
10. Male derived s t a t i s t i c s , Group I I ...... 45
11. Female derived statistics, Group II ...... 46
12. Male measurements, Group I I I ...... 47
13. Male derived s t a t i s t i c s , Group I I I ...... 47
14. Female measurements, Group I I I ...... 48
15. Female derived statistics, Group III ...... 48
16. Infection of snails with Cosmocercoides dukae from amphibians ...... 61
v i i TABLES (Contd.)
T ab le Page
17. Infection of snails with Cosmocercoides dukae by ingestion of frog fe c e s ...... 63
18. A comparison of the species of the genus Cosmocercoides ...... 70
v i i i INTRODUCTION
The nematode Cosmocercoides dukae (Cosmocercidae) has been found in a variety of hosts. In fact, the great diversity of animals that apparently serve as definitive hosts raises some doubt whether all such reports are con cerned with Cosmocercoides dukae or whether some of the specimens that have been assigned to this species may belong to a closely related one which has not been defined taxonomically. It must be admitted that a species such as
C. dukae which has been reported from such diverse hosts as molluscs, as well as such vertebrates as amphibians and reptiles, is either very versatile in its acceptance of host animals or many closely related species of nematodes may be involved. On the other hand, since specimens of C. dukae from the same species of host exhibit such great variation in size and shape, it might be that there are fewer valid species in the genus Cosmocercoides than are now accepted, and that those species whose descriptions have been based on relatively few specimens may actually be synonyms of C. dukae.
An attempt was made to identify specimens of what appeared to be Cosmocercoides dukae from the American Toad,
Bufo americanus, and from the Eastern Spadefoot Toad,
Scaphiopus holbrooki. Specimens from the former host were larger than those from the latter, and, seemingly more
important, the nematodes from Bufo americanus contained more papillae than those from Scaphiopus holbrooki. Since vari
ation in the number of papillae is often a significant
factor in the classification of nematodes, the writer felt
that it would be of value to examine specimens from other
species of hosts in order to compare the number of papillae
as well as other factors. In the original description of
this species from the newt, Triturus viridescens, exactly
twelve pairs of complex papillae were listed by Holl (1928).
Although several authors noted that C. dukae exhib
ited marked variation in size and shape, no one had
attempted to define the degree of variability.
In addition, it has never been determined whether
molluscs could become infected by acquiring the eggs or
larvae of C. dukae taken from amphibian hosts.
A study was made (1) to determine the types of
cosmocercid nematodes in amphibians and reptiles in Ohio;
(2) to describe the degree of variability of C. dukae in
Ohio; (3) to relate and compare these nematodes to species
of the benus which are found elsewhere; and (4) to see
whether C. dukae from amphibians will infect molluscs. In
order to accomplish these objectives, several lines of
investigation have been followed. First, wherever possible,
loans have been obtained of type specimens to check against 3 published descriptions. Second, specimens have been obtained from post-mortem examinations of animals that already have been reported as hosts of C. dukae. Some of these
n " specimens were then checked against the descriptions already given fo r members o f the genus. Others were used as a source of eggs and larvae in attempts to infect molluscs. HISTORICAL REVIEW
The genus Cosmocercoides belongs to a group of round worms commonly referred to as oxyuroids. All species of oxyuroids possess an esophageal bulb set off from the pre ceding portion of the esophagus by a constriction. The name
Cosmocercoides apparently stems from the word Cosmocerca, a related genus erected by Diesing in 1861.
Wilkie (1930) justified the erection of the genus
Cosmocercoides when he noted the lack of subcuticular supportive structures associated with the plectanes that are characteristic of the genus Cosmocerca. He named two species, viz., Cosmocercoides pulcher and C. trldens. The first, from Bufo japonicus, was indicated as the type species of the genus. Unfortunately, Cosmocercoides tridens was described from only one specimen, a male, taken from Tylotriton andersoni in Japan.
Travassos (1931) accepted Wilkie's genus, placing
Cosmocerca dukae Holl, 1928 in the genus Cosmocercoides.
In the same year Travassos also placed Oxysomatium variabilis Harwood, 1930 in the genus Cosmocercoides.
Karve (19^4) added still another species by describing
Cosmocercoides bufonis from the toad Bufo himalayanum in
In d ia. S k rjab in , Schikhobalova, and Mosgovoy (1951) 5 placed Cosmocerca skrjablnl Ivanitsky, 19*10, which was
obtained from Rana temporaria in Russia, in the genus
Cosmocercoides. Another species, Cosmocercoides m u ltip a p illa ta , was described by Khera (1958) from
Bufo melanostlcus in India. Cosmocercoides v a r ia b ilis was placed in synonymy with Cosmocercoides dukae by Ogren (1953). Although
Yamaguti (1961) and Skrjabin et a l. (1961) agreed to this
arrangement, Chabaud and Brygoo (1958) and Anderson (i 960 ) were re lu c ta n t to accept th is synonymy since th ere were
differences between the account of the life cycle of
Cosmocercoides v a r ia b ilis given by Harwood (1930) and th a t
given by Ogren (1953; 1959 a and b ) .
Skrjabin et a l. (1951) added Cosmocerca timofejovoi
Skarbilovich, 1950 to the genus Cosmocercoides. Skrjabin
apparently felt that this species did not have plectanes
with subcuticular supportive rods. However, it agrees in
other respects with the genus Cosmocercoides.
There is uncertainty at present concerning the
validity of Trionchonema rusticum Kreis, 1932. Chitwood
(1933a) placed this species in the genus Cosmocercoides, but
he did not state why this was done. Although Kreis'
drawings and measurements agree in many respects with the
known species of Cosmocercoides, the three "spears" which he
shows within the non-bulbous portion of the esophagus are
definitely different, as are also the gland-like, spindle shaped pre-anal papillae. Kreis (1932) did not report the small papillae which occur throughout the total length of the cosmocercid body wall. Skrjabin (1961) accepted the work of Kreis, but included Cosmocercoides rusticum (Kreis,
1932) Chitwood, 1933 as a synonym of Cosmocercoides dukae.
Yamaguti (1961) placed Trionchonema K reis, 1932 in synonymy with Cosmocercoides Wilkie, 1930. Ballesteros-Marquez
(19^5) in his revision of the Cosmocercidae, placed
Cosmocercoldes rusticum in synonymy w ith Trionchonema rusticum. Kreis found this nematode in the land snail
Polygyra espicola Bland in City Park, New Orleans, Louisiana.
There have been other reports of cosmocercid nema todes in land-dwelling gastropod molluscs. Anderson (i 960 ) noted that Barthelemy in 1858, in naming Ascaroides limacis, as well as Conte and Bonnet in 190 3, in naming Angiostoma helicis, were probably dealing with species of the family
Cosmocercidae. However, not enough data were given to determine whether these forms belonged to the genus
Cosmocercoides. Chitwood and Chitwood (1937a) regarded
Angiostoma helicis as a cosmocercid from the snail Helix.
Chitwood (1933b) indicated that Leidy's Ascaris cylindrica
(Angiostoma cylindricum) belonged to the genus
Cosmocercoides.
Ogren (1953) obtained C. dukae from the land snails
Ashmunella rhyssa edentata, Polygyra fosteri, and
Retinella sp. as well as from specimens of the slug, Deroceras sp. Anderson (i 960 ), working with two species of land snails, Discus cronkheitei and Zonitofdes arborea in
Ontario, Canada, found cosmocercids which he tentatively identified as Cosmocercoides dukae. He also found speci mens of this same worm in the slug Deroceras gracile as well as in the amphibians Rana pipiens, R. clamitans,
R. catesbeiana, R. septentrionalis, Bufo americanus, and
Ambystoma jefferso n ian u m . Harwood (1930) found th a t
Oxysomatiurn variabilis, now a synonym of Cosmocercoides dukae, was oviparous; however, both Ogren and Anderson have subsequently given evidence that C. dukae was ovoviviparous.
Anderson observed that when females of C. dukae were removed from molluscs, they usually discharged part of the contents of their uteri regardless of whether the eggs were in an advanced stage or were poorly developed. He found that eggs with first stage larvae usually hatched and developed to the infective stage in tap water, while younger embryos failed to develop. Anderson also felt that
Harwood's infection experiments would have to be confirmed by fu rth e r work.
Harwood (1930) Introduced in fe c tiv e larvae o f C. dukae into amphibians in three ways, viz., (1) directly on the skin; (2) by subcutaneous injections; and (3) orally.
In certain cases he found larvae, presumably Cosmocercoides, in the lungs, but unfortunately drawings were not included in his report. However, Kung and Wu (19^5) found adults of Cosmocercoides pulcher in the large intestine of Bufo bufo japonicus. They also found larvae of apparently the same species in the lungs of the same host. Their identification of the larvae was based on the general appearance of the worm, and especially on the arrangement of structures of the genital system.
Anderson's experiments, in infecting amphibia, in volved feeding snails that were infected with C. dukae to
Rana pipiens or in pipetting the infective third stage larvae of the worm into the esophagus of this frog. Both larval and adult forms were passed with the feces of the frogs over a period of a few weeks. Worms rarely were found on autopsy after thirty days. Prom Anderson's results there was no evidence that C. dukae could establish itself in amphibians,
Chabaud and Brygoo (1958) preferred provisionally to keep the name Cosmocercoides variabilis for the material studied by Harwood, since it seemed possible to them that
C. dukae and C. variabilis might be two different species.
Further life history study is needed, especially with specimens obtained from amphibians, in order to reveal whether these two species are distinct or whether they will have to be placed in synonomy. There can be little doubt that C. dukae is a well-established parasite of land snails and slugs, especially since it has been shown that, the life
cycle can be completed in two ways. Both Anderson (i 960 ) and Ogren (1953) have shown that Infective larvae, having left one snail, can enter the mantle cavity of a new snail host and continue the cycle. In addition, Anderson (i 960 ) has described the transmission of larvae by means of snail and slu g eggs, whereby in fe c tiv e netmatode larvae in the genital tract of the adult mollusc became trapped within the egg shell of the young developing mollusc. Later, such larvae apparently penetrated the mantle cavity of the embryo mollusc before it hatched. Most certainly this insures a high percentage of infection.
There are differences of opinion concerning the classification of the genus Cosmocercoides. Travassos
(1931) placed it in the family Cosmocercidae Travassos,
1925, which is included in the super-family Subuluroidea
Travassos, 1930. Yamaguti (1961) retained the family
Oxyuridae Cobbold, 1864, considering Cosmocercoides a member of the subfam ily Cosmocercinae R a i l l i e t , 1916 on the basis of (a) the presence of a gubernaculum and two equal spicules in the male and (b) anteriorly located ovaries in the female. Ballesteros-Marquez (1945) followed the scheme of Travassos. Skrjabin and Schikhobalova (1951) removed the family Cosmocercidae from the superfamily Subuluroidea and erected the superfamily Cosmocercoidea, with Cosmocercidae as the type family. B. G. Chitwood (1937) placed the family
Cosmocercidae in the superfamily Ascaridoidea (Railliet and
Henry, 1915) along with the Kathlaniidae, Heterakidae and 10
Ascarididae. The Oxyuridae, Thelastomidae, Rhigonematidae,
and Atractidae were assigned to the superfamily Oxyuroidea.
The Ascaridoidea possess the ventro-lateral cephalic papilla near the amphids, whereas this papilla is missing in the
Oxyuroidea. B. G. Chitwood provided Thorne (1959) with an
outline of higher nematode groups in which he endorses the
superfamily Cosmocercoidea Skrjabin and Schikhobalova, 1951.
These differences of opinion with respect to the higher taxa may result, in part, from disagreement among nematologists concerning the amount of data which should be obtained prior to revising various groups. M. B. Chit wood (personal communication) indicates that much informa
tion concerning cosmocercid nematodes will be required in
order to clarify their systematics.
Skrjabin et al . (1961), listed the following
genera under the subfamily Cosmocercinae: Cosmocerca Diesing,
1861 (type genus); Paracosmocerca Kung and Wu, 1945;
Cosmocercella Steiner, 1924; Cosmocercoides Wilkie, 1930;
and Trionchonema Kreis, 1932. All of these genera are com
posed of species exhibiting plates (either plectanes or
false plectanes) on the ventral surface of the tail of the
male specimen. MATERIALS AND METHODS
Specimens of Cosmocercoides dukae were obtained p ri marily from hosts collected by the writer in Delaware and
Hocking Counties in Ohio during the spring, summer, and fall of 1963. However, collections were made also from other locations at this time (Table 1). Nematodes furnished by the
U.S. National Museum were from hosts in Wyoming, Florida, and
North Carolina. This included the holotype of C. dukae c o lle c te d by H oll in 1928 In North C arolina. The B ritis h
Museum (Natural History) furnished cotypes of Cosmocercoides pulcher, and also sent additional requested information con cerning Cosmocercoides tridens which had been described from a single male specimens
Dr. Paul D. Harwood sent specimens of C. dukae col lected by him in Houston, Texas. Many specimens of C. dukae from hosts taken in twelve collecting sites in Ohio were received from Dr. John L. Crites of the Department of
Zoology and Entomology of the Ohio State University.
One hundred alimentary tracts of the North Carolina salamander, Plethodon longicrus Adler and Dennis, 1962, were furnished for examination by Dr. Barry Valentine of the same department, and specimens of C. dukae were recovered from them.
11 12
W. A. Reid of the University of Kansas loaned speci mens of C. dukae which were taken from Rana catesbeiana in
North Carolina.
Amphibians were collected by hand or with a dip net by the writer during daylight hours. At night, frogs were
approached slowly with a bright light and caught by hand.
Larger numbers of frogs were transported in minnow buckets, while salamanders fared well in a moist "bait canteen" con
taining moss or leaf litter. When the collection of large
numbers precluded immediate examination, host animals were
stored in an ordinary household refrigerator at 5 C.
Amphibian hosts were examined as soon as possible
a f t e r being c o lle c te d . They were k ille d by d e c a p ita tio n or
by "pithing" the brain and spinal cord. Organs and areas
examined included the large intestine, small intestine,
stomach, lungs, liver, urinary bladder, and coelom. These
sites were checked individually in watch glasses or petri
dishes, both grossly and with the aid of a dissecting
microscope.
Snails of the species Subulina octona obtained from
the greenhouse of the Department of Biological Sciences of
Plattsburgh State University College, Plattsburgh, New
York, were cultured at laboratory temperatures of 24 - 28 C.
from November 1966 through the summer of 1967. Pour large
snails with shells about 16 mm long were placed in a covered
petri dish containing a paper towel saturated with 13 distilled water. They were fed lettuce, and given chalk as a calcium carbonate source as suggested by Anderson (i 960 ).
Young snails were removed from such brood chambers weekly to facilitate cleaning. At this same time both dishes and snails were checked to determine whether the cultures were nematode-free.
Nematodes were usually introduced to snails by pipetting them onto the withdrawn foot. When the nematodes disappeared from sight the snail was isolated in a covered
Syracuse watch glass and checked for two to four hours to determine whether entrace had been accomplished. Subse quently, watch glasses containing a snail, lettuce, chalk, and several drops of water were stored in moist containers.
Moist toweling or filter paper was not placed in the watch glass since it was felt that nematodes might be lost in it.
To determine whether snails could be parasitized by in gesting the feces of infected frogs, snails which had been without food for two days were fed pieces of lettuce, about
10 mm. x 10 mm., upon which feces had been placed.
Living nematodes were examined in saline with their movement re ta rd e d by the pressure o f a cover s l i p . F ixation was accomplished by (a) placing the worms in a Syracuse watch glass in 3 to 5 ml of Ringer's "cold-blooded" saline and (b) quickly adding to this container 10 to 15 ml of 50 per cent ethanol which had been heated to 70 C. Worms were cleared in a mixture of glycerine and 75 per cent ethanol in the proportion of 1:9. The dish was placed in a dust-free container, and the alcohol was then allowed to evaporate.
Some specimens were cleared in lactophenol; others in phenol to which a small amount of Semichon's stain had been added. Temporary mounts were made either in glycerine, or in glycerine jelly according to the suggestions of Cobb
(1920). En face views were made by using the techniques of
Anderson (1958) . This consisted of placing the specimen in glycerine-jelly on a cover slip, after which decapitation was accomplished with a razor blade. The jelly was then melted by warming the coverslip, the anterior tip of the specimen oriented to touch the cover slip, and the jelly allowed to harden by cooling. The cover slip was then placed on a s lid e and was supported by two sm all c a p illa ry tubes which held the specimen in position. Finally, the cover slip was sealed around the edge with either vaseline or glycerine jelly. This temporary .mount was then examined under an oil immersion lens. Drawings were made with the aid of either a camera lucida or a micro-projector.
Measurements were made with an ocular micrometer scale. OBSERVATIONS AND RESULTS
Occurrence of Cosmocercoides dukae
Prom A pril 1963 to July 1964, a t o t a l of 295 anim als, representing fifteen species of amphibians and three species of reptiles, were examined as possible hosts of Cosmo cercoides dukae. Of the fifteen species of amphibians, 288 individuals, representing fourteen of the fifteen species, were collected from sixteen sites in ten Ohio counties
(Table 1). Of these 288 individuals, 37 (19.8$), represent ing ten different species of hosts, harbored C. dukae. The fifteenth species, one individual of the Mountain Spring
Salamander, Gyrinophllus danielsi was not parasitized. This latter species, in addition to one specimen of the Northern
Ringneck Snake, Diadophis punctatus edwardsi, also not parasitized, was collected in Greene County, Pennsylvania, and given to the writer for examination by Dr. Russell R.
Williams of Waynesburg College.
In addition to the Northern Ringneck Snake, one specimen of the box turtle Terrapene Carolina from Carter
County, Kentucky, and four specimens of the garter snake
Thamnophis sirta lis from Calamus Swamp, Pickaway County, were uninfected with C. dukae TABLE 1
COSMOCERCOIDES DUKAE FROM OHIO AMPHIBIANS, APRIL 1963-JULY 1964
Host Location Number Number Worms per Infected Host Examined Infected Mean Range
Acris crepitans Wayne Twp., Pickaway Co. 11 0 _ • Laurel Twp., Hocking Co. 1 0 - - (Cricket frog) Pickaway Twp., Pickaway Co. 1 0 -- 13 0
Bufo
B. americanus W estfield Twp., Morrow Co. 3 0 Laurel Twp., Hocking Co. 16 15 (American toad) Genoa Twp., Delaware Co. 1 0 Benton Twp., Pike Co. 4 3 24 18 (7 550 22 1 to 140 B. woodhousei fow leri Brown Twp., Vinton Co. 1 1 1 1 (1002) 13 — (Fowlers Toad)
Desmognathus fuscus Colerain Twp., BeLmont Co. 9 0 Berne Twp., F a irfie ld Co. 1 0 (Dusky salamander) Laurel Twp., Hocking Co. ■ 33 4 Goodhope Twp., Hocking Co. 13 0 Clear Creek Twp., F a irfie ld Co. 3 0 1 Pickaway Twp., Pickaway Co. 1 0 1 60 4 (72) 1 0 Table 1 (contd.)
Host Location Number Number Worms per Infected Host Examined Infected Means Range
Eurycea
n E. b islin e a ta Norwich Twp., Franklin Co • -L . 0 Colerain Twp., Belmont Co 1 0 (Two-lined salamander) Benton Twp., Pike Co. 1 0 Laurel Twp., Hocking Co. • 4 2 7 2 (292) 1 0 E. longicauda Laurel Twp., Hocking Co. 1 0 (Long-tailed salamander)
Plethoaon glutinosus Benton Twp., Pike Co. 1 0 - • (Slimy salamander)
Pseudacris brachyphona Laurel Twp., Hocking Co. 6 5 6 3 2 to 6 (Mountain chorus frog) 5 (83£)
Ran a R. catesbeiana W estfield Twp., Morrow Co. 110 ' 15 Berne Twp., Fairfield Co. 3 0 (Bullfrog) Genoa Twp., Delaware Co. 3 0 Etna Twp., Licking Co. 1 0 Benton Tv,'p., Pike Co. 2 0 Concord Twp., Delaware Co 1 0 120 15 (12$) 4 1 to 16 Table 1 (contd.)
Host Location Number Number Y/orms per Infected Host Examined Infected Means Range
Rana - R. clamitans Norwich Twp., Franklin Co. 1 0 Berne Twp., F a irfie ld Co. 2 0 (Green frog) Genoa Twp., Delaware Co. 1 0 W estfield Twp., Morrow Co. 1A A Benton Twp., Pike Co. 10 1 i Wayne Twp., Pickaway Co. 5 0 33 5 (1556) 5 1 to 12 R'. pipiens Ysestfield Twp., Morrow Co. 5 0 Newton Twp., Licking Co. 1 1 (Leopard frog) Wayne Twp., Pickaway Co. 2 0 { 8 1 {12%) , .6 i 1 i R. sylvatica Laurel Twp., Hocking Co. 7 5 t t Benton Twp., Pike Co. J_ 0 (Wood frog) s 5 (63 %) 2 1 to A
Scaphiopus holbrooki Berne Twp., F a irfie ld Co. 1 1 1 1 (100%) 7 (Spadefoot toad)
Triturus viridencens Goodhope Twp., Hocking Co. 5 0 -
(Spotted newt) a 19
In summary, a total of 57 (19*3$) of the 295 animals
examined harbored Cosmocercoides dukae.
It must be noted that one collecting site in Laurel
Township in Hocking County was visited more often after it was found to be a good source of infected hosts. A total
of 29 (48$) of the 60 animals collected in Crane Hollow in
Laurel Township contained C. dukae. The percentage of
infection in Ohio amphibians which were examined in this
study, other than those from Laurel Township, was 11.9 per
cent. Thus the percentage of infection (19.3$) observed in -
Ohio amphibians from the localities indicated in Table 1 is probably somewhat higher than it would have been if the number of host specimens examined from these various locali
ties had been equally large.
In the present study the highest percentage of infec
tion with Cosmocercoides dukae was found in Pseudacris brachyphona (83$), followed by Bufo americanus (75$), and
Rana sylvatica (63$). Although no records of percentage of
infection for Rana sylvatica were found in the literature, high percentages have been reported by other workers for
certain species of Bufo and Pseudacris. Brandt (1936) found
78 per cent infection in Pseudacris brimleyi from Beaufort
County, North Carolina, while Harwood (1930) listed 86 per
cent in Bufo valliceps, and 58 per cent in Pseudacris
triseriata from the vicinity of Houston, Texas. 20
In the present study, somewhat lower percentages of infection were found in Rana catesbeiana (12$). R. clamitans (15%), and R. pipiens (12$). Other workers have reported percentages ranging from twelve to forty for these species of amphibians.
In addition to specimens of Cosmocercoides dukae obtained from post mortem examinations, nematodes specimen from other sources were identified and examined in the present study. Cosmocercoides dukae from seven species of hosts were loaned to the writer by Dr. John L. Crites of the Department of Zoology and Entomology at Ohio State Uni versity. Host records for C^. dukae in Ohio which were obtained from Dr. Crites include Pseudacris triseriata,
Plethodon glutinosus, and Ambystoma opacum. The description of C. dukae from Ohio was based on a morphological study of nematodes from these three hosts and also from the ten infected species collected by the writer (Table 1). A sup plementary group of nematodes was also received from Dr.
Crites. These specimens of C. dukae were recorded from
Rana clamitans, R. pipiens, and Eurycea bislineata, host species also found infected by the writer.
The Morphology of Cosmocercoides dukae from Ohio Amphibians
The following description of C_. dukae is based on specimens obtained from Amphibia in Ohio. The measurements given were taken from mature individuals. In the case of 21 female worms, those that contained at least a few eggs were considered to be mature. Twenty-seven of the thirty-nine male specimens studied were obtained from Bufo americanus.
The remaining twelve males had been taken from eleven other different host species. Twenty-five of the thirty-four female specimens studied were obtained from B. americanus, and the other nine females were from nine other host s p e c ie s .
Differences between specimens of Cosmocercoides dukae taken from different host species were noted early in this study. Although size differences alone would not seem to be of critical importance, variation in the number and arrange ment of papillae are often of diagnostic value in nematode classification. As the literature concerning C. dukae was examined, it became apparent that many studies had omitted certain important morphological details, and therefore recording observations on specimens from Ohio amphibians as completely as possible would be of value to anyone doing resarch with cosmocercid nematodes.
C uticle
Cuticular striations ring the nematode throughout its entire length in a plane perpendicular to the long axis of the body. These striae are not seen adjacent to, or on either side of, the lateral alae. However, in several instances fine indentations are seen along an ala as though 22 the striations were continuous. A coarsely striated secre tion which Harwood (1930) observed coating the finely striated cuticle was never present in specimens of C. dukae obtained from Ohio amphibians. This secretion may have represented incompletely molted cuticle, since Harwood observed that the secretion also contained fine striations.
The distance between the striae at mid-body varies from 2.4 to 6.4 microns in males and from 2.4 to 8.0 microns in females.
Very minute body papillae, or micropapillae, are found scattered over the entire cuticle of specimens of
C. dukae from Ohio amphibians. These papillae begin anteriorly 25 to 35 microns behind the mouth and end pos teriorly in the vicinity of the phasmids. Their position is not random, since they are arranged on each side of the specimen in six longitudinal rows which are most easily discerned in the region between the esophageal bulb and the anus. Thus on each side there are three rows dorsal and three ventral to the lateral alae. The most ventral of these rows on either side, slightly below a ventro-lateral position, is in line with a row of complex papillae. Although occasionally a micropapilla may be seen between two rosettes, the ventral row appears to be replaced by the rosettes in this region. In the post-anal region of the males, both micropapillae and larger simple papillae are found, while in females the post-anal region contains only micropapillae 23
(Pigs. 17, 22). Khera (1958) found only minute papillae on
the tail of the female of Cosmocercoides m ultipapillata, but made no mention of differences among simple papillae on
the tail of the male.
A single nerve fiber is visible, connected to each micropapilla in favorable specimens. The surface of the papillae is only slightly rounded.
Cephalic region
The lips of larger specimens, which can be viewed
easily, measured eight microns in length (Pig. 6). The
papillae and boundaries of the lips, when seen in an en face
view, are extremely difficult to study. The inner boundary
of each of the th re e lip s appeared as shown by Anderson
(I960) for Cosmocercoides dukae which he obtained from
Ontario molluscs, but the postero-lateral boundaries were
not seen. Anderson showed the outer circle of papillae on
the lips (Pig. 31). On the other hand, Ogren (1953) illus
trated one pair of papillae on each of the ventro-lateral
lips and two pairs on the margin of the dorsal lip (Fig.
30). These four pairs of marginal papillae represent
Anderson's four horseshoe-shaped papillae. Anderson was not
sure that these horseshoe-shaped papillae should be regarded
as double, since only one or two of those that he observed
gave him that impression. The marginal papillae in the 24 present study appear U-shaped or horseshoe-shaped, but
rarely appear double (Pig. 28). On the other hand,
Chitwood and Chitwood (1937) Illustrated each of these
four papillae as double.
Esophagus
The anterior tip of the esophagus, termed pharynx by
some authors, has an appearance distinct from the posterior
portion, possibly because of a difference in the orienta
tion of the muscle fibers. In an en face view, the anterior
end of the esophagus terminates in three bluntly pointed
projections immediately beneath and median to the lips. As
Anderson (i 960 ) suggested, this positioning gives these
projections the appearance of teeth.
The ducts of the esophageal glands were not seen;
however, refractile granules at the posterior boundary of
the pharynx, and also in a position about 50 microns anteri
or to the esophageal bulb, indicate the regions of the
gland openings (Pig. 6). The valved bulb agrees in form
with that of C/ dukae drawn by other investigators (Fig. 11).
When the bulb is contracted, as indicated by the valve
apparatus which is located anteriorly within the bulb, cells
at the posterior end protrude into the intestinal lumen.
These cells, making up the esophago-intestinal valve, do not
usually protrude posteriorly in preserved nematodes, but
they are seen distinctly in living material. These cells are
well-developed in the larval stages. 25
In te s tin e
The intestine is a straight, uncoiled tube throughout its length. At its beginning it is slightly wider than the esophageal bulb, but it gradually reduces to about one-half this width by mid-body. In a few specimens the shape of
the cells of the intestinal wall was visible, appearing
irregularly polygonal with five, six, or seven sides
(Fig. 27).
Rectum
The intestine ends in an easily seen clump of cells
called the intestino-rectal valve (Fig. 1). The remaining portion of the alimentary canal, lined with cuticle, is
termed rectum in the female and terminates with the anus
(Fig. 22). The latter is a ventral, transverse slit (Fig.
21). In mature females, the rectum ranged in length from 80
to 136 microns. Since this region in the male receives the
contents of both the intestine and the reproductive tract,
it is called the cloaca (Fig. 1). In mature males it is
from 80 to 128 microns long.
Simple caudal papillae
Four pairs of simple caudal papillae are consistently
present on males of C. dukae (Fig. 17). Two pairs are
located posterior to the phasmids at approximately 40 microns
from the tip of the tail. One of these pairs extends dorso-
laterally, and the other ventro-laterally at about the same 26
level. The other two pairs of caudal papillae are located anterior to the phasmids at approximately 70 microns from the tip of the tall. One of these pairs extends dorso- laterally and the other, ventro-laterally. Extra papillae were found on some specimens, but were irregularly spaced and often unpaired.
The simple median papilla on the anterior cloacal lip of C. dukae was a double papilla in nearly every male examined (Pig. 17). This papilla is based on a rounded flat cone which in ventral view appears twice as wide as long. In lateral view this papilla and its base give the appearance of a mammilation. The base appears to be cuti- cularized. This papilla is not represented in the female specimen. Ogren (1953) referred to only one precloacal papilla which he said was indistinct. Anderson (i 960 ) found a distinct precloacal papilla on male specimens of
C. dukae from Ontario molluscs.
Female reproductive system
The female reproductive system of Cosmocercoides dukae consists of two ovaries, two oviducts, two seminal receptacles, two uteri, a vagina, and the vulva (Figs. 23 and 19). The configuration of the female reproductive tract is followed most easily within individuals containing relatively few eggs. In older females that are filled with hundreds of eggs, it is difficult to locate the junction of 27 the two uteri with the vagina. This junction may shift
from the level of the vulva where it is found in young mature individuals. A small ring of cells encircling the
vagina may function as an ovejector or as a sphincter, but
this was not determined in this study (Pig. 19). The walls
of the vagina are not of equal thickness throughout its
length (Pig. 23).
The female reproductive tract was removed quickly
from a living worm taken from an American toad, placed in
saline, and measured immediately. The first portion of
the vagina adjacent to the vulva, which contains a very
small lumen, measures 703 microns in length and 56 microns
in width. The thinner walled portion which has a larger
lumen, extends to the origin of the two opposed uteri. This
portion measures 199 microns in length, thus giving a total
length of 902 microns fo r the vagina. The w alls of the
thin-walled portion were similar in thickness to the
uterine walls (Pig. 23). The thick walled portion of the
vagina is probably important in the process of the ejection
of ova or larvae. According to Chitwood and Chitwood
(1937), the entire portion from vulva to uteri should be
termed vagina, if it is lined with cuticle. Verification
of this for C. dukae will require histological study. This
same section of reproductive tract in the related species,
C. m u ltip a p illa ta , as shown by Khera, is also composed of
two distinct regions. For slightly more than one-third of 28 its length, adjacent to the vulva, the walls of the vagina of C. multipapillata are thick, as in C. dukae. The remain der is thin walled. Khera (1958) named this portion of the reproductive tract, in C. multipapillata, the uterine tube. This region in C. dukae might also be called vagina uterina, but again confirmation would depend on a histolog ical study to determine whether it is lined with cuticle *
In discussing the reproductive system of nematodes, Chitwood
(1937) states that it is difficult to discover the exact limits of the vagina by gross study.
The uteri of older females of C. dukae contain eggs in various stages of development (Pigs. 33-39). Polar bodies were often observed in eggs containing cleavage stages (Pig. 3*0.
The seminal receptacles, or spermathecae, marking the ovarian ends of the uteri, are usually easily visible as avoid expansions of the reproductive tract Pig. 23).
They become more d i f f i c u l t to lo cate in vivo as they lose their stored spermatozoa and as the body cavity becomes filled with the egg-laden uteri. When the reproductive tract is removed from the worm, the region of the recep tacles may still be seen as an avoid expansion even in older females with the sperm supply depleted.
Male rep ro d u ctiv e system
The reproductive tract of the male extends anteriad for about two-thirds to four-fifths of its length. Behind 29 the esophagus the testis bends posteriorly, gradually narrowing towards its distal end (Pig. 5). The tract is thin-walled with the exception of the ejaculatory duct which empties into the cloaca (Pig. 1). This duct joins the cloaca in a position ventral to the intestine.
Special male anatomy
The degree of prominence of the cloacal lips varies in different specimens. These lips appear less prominent in living specimens than in preserved ones. Contraction of the strong muscles in the cloacal region during fixation possibly affects cloacal lip configuration.
Measurements were made of the diameter of two rosettes on male specimens of C. dukae from Ohio amphibians
(Pig. 8). Posteriorly, one of the last four rosettes, and anteriorly, one of the rosettes from twelve to sixteen was chosen for measuring. The choice of which rosette was to be measured was based merely upon, which ones were oriented in a plane nearly perpendicular to the line of sight.
Anterior rosettes are usually larger in diameter tahn those located posteriorly. The diameter of the anterior rosettes is approximately one and one-half times greater than that of the posterior rosettes.
A study of the rosettes on nineteen worms revealed twelve to twenty tubercles and eleven to nineteen micro tubercles surrounding the central papilla (Pig. 8). The use 30
of an oil immersion lens was usually required to count the microtubercles.
Descriptive Summary of Cosmocercoides dukae from Ohio Amphibians
General
Anterior end with three lips. Six minute papillae
in the inner circle, two on each lip (Pig. 28). Outer
circle consisting of four U-shaped papillae, two on the
dorsal lip, and one ventrally on each ventro-lateral lip.
Flat, ventro-lateral papillae located laterally on ventro
lateral lips. Minute amphids located laterally just beyond
outer circle. Esophagus with posterior bulb, slight isthmus,
followed by long, slightly tapering cylindrical corpus;
division into procorpus and metacorpus indistinct. Cuticle with uniform transverse striations interrupted laterally by
very narrow inconspicuous lateral alae, the latter extend
ing from the level of the anterior fourth of esophagus to
level of cloacal opening in male, and nearly to caudal
extremity in female. Numerous tiny papillae over entire
cuticle in six nearly regular longitudinal rows on each
side of body. Tail finely pointed.
Male (P igs. 1, 5)
Length, 1.98-5.30 mm; width, at mid-body, 146-452
microns. Esophagus, 0.4Q-1.00 mm long; portion anterior
to opening of dorsal esophageal gland 40-78 microns long; 31 bulb portion, 70-139 microns in diameter. Nerve ring,
162-371 microns from anterior end. Excretory pore,
252-570 microns from anterior end. Spicules, equal,
arcuate; 221-478 microns long. Gubernaculum, triangular in
outline; 90-169 microns long. Tail, 104-209 microns long, with four pairs of simple papillae constantly present, two pairs approximately 40 microns from caudal tip and two pairs 30 microns anterior to them. Phasmids, 56-72microns
from end of t a i l . Complex p a p illa e , lo cated mainly in two
ventro-lateral rows on posterior third of body; last three
on each side arranged in triangle in cloacal region; posterior pair always post-cloacal; next two pairs ad-anal,
one dorsal to other. One double papilla on anterior clo
acal lip, on heavily cuticularized base. Spermatozoa,
spheroidal, 6-8 microns in diameter. Minute papillae
separated linearly by 90-188 microns. Cuticle striations
separated by 2.4-8 microns.
Female (Fig. 22)
Length, 2.92-6.12 mm; width, 133-418 microns.
Esophagus, 0.44-1.02 mm long; portion anterior to opening
of dorsal esophageal gland, 48-72 microns long; bulb por
tion, 83-152 microns in diameter. Nerve ring, 192-351
microns from anterior end. Excretory pore, 285-597 microns
from anterior end. Vulva, a transverse slit behind middle
of body; 1.73-3.34 mm from anterior end. Vagina directed 32 anteriorly for first 176-365 microns, then posteriorly for same distance. Opposed uteri lead from vagina to sperma- thecae; anterior ovary and oviduct directed posteriorly for one-half to three-fourths of their length; posterior oviduct and ovary extend anteriad from spermatheca, end ing posterior to level of esophageal bulb. Phasmids lo cated on lateral alae, 80-90 microns from pointed tip of tail. Tail length, 160-305 microns. Eggs symmetrical;
44-93 microns long by 25-46 microns wide.
Amphibian Hosts in Ohio
Bufo americanus; B. woodhousei fowleri; Scaphiopus holbrooki; Eurycea bislineata; Rana sylvatica; R. clami tans, R. catesbeiana; R. pipiens; Bseudacris brachyphona;
P. triseriata; Plethodon glutinosus; Ambystoma opacum; and
Desmognathus fuseus.
Site of infection
Anterior end of large intestine.
L o c a litie s
Fairfield, Franklin, Hocking, Morrow, Sandusky, and
Vinton Counties in Ohio. 33
Observations on the Holotype of Cosmocercoides dukae (Holl, 1928)
Certain structural details observed in specimens of
C. dukae from Ohio amphibians were not included by Holl
(1928) in his original description of this species.
However, Holl's specimens were taken from the large intes tin e of a newt, T ritu ru s v irid e s c e n s , from Durham, North
Carolina. Since a better knowledge of the type specimen would allow fuller comparisons to be made, this specimen was located and studied. It had been deposited by Holl on a slide in the Helminthological Collection of the U.S.
National Museum. Unfortunately, most of the mounting medium had evaporated, leaving the specimen in a hardened coat of balsam. With permission, the specimen was removed and remounted after washing in xylene.
Measurements of spicule, gubernaculum, and body length agreed closely with Holl's figures. However, body width, tail length and dimensions of the esophagus varied greatly, probably caused by the distorting effects of the evaporation of the mounting medium. The original figures are probably more accurate. The nerve ring and excretory pore are 0.143 and 0.238 microns, respectively, from the anterior tip. These were not mentioned by Holl. Lateral alae, also not mentioned originally, extended on the holo type from a point 70 microns behind the anterior tip of the worm to about 23 microns in front of the level of the 34 anus. The alae could be traced throughout their entire length in spite of cuticular wrinkling.
Holl recorded eleven pairs of pre-anal and one pair of post-anal complex papillae which he termed circular plectanes. Although some were partially obscured by dried balsam, there appeared to be ten or eleven pairs arranged in two rows. There also appeared to be a pair of ad-anal complex papillae located slightly dorsal to these rows as
Wilkie (1930) and most other investigators have observed.
Simple caudal papillae and micropapillae also were observed on the type specimen (Pig. 2). There is a single median, simple papilla on the anterior cloacal lip similar to that seen on C. dukae from other areas of North
America.
Although three lips are visible at the anterior tip of the worm, papillae are not visible. This is probably due to the effects of the drying during storage on the slide.
Comparison of Selected Groups of Cosmocercoides dukae
Certain groups of specimens were compared in order to define the degree of variability among nematodes assigned to the species Cosmocercoides dukae.
A single host specimen of Bufo americanus from Laurel
Township in Hocking County yielded 130 individuals of
Cosmocercoides dukae. Most of these worms were apparently recently matured since the number of eggs per female 35 ranged from approximately six to forty as compared to hun dreds of eggs observed in worms from other host individuals.
The distances of fixed points, e.g., the excretory pore, were measured from the anterior tipof 48 of the 130 worms. The p o s itio n of the vulva was also measured from the extreme posterior tip of the body.
When measurements from this series of 48 specimens were compared with those given by Anderson (I960) for twenty male and ten female specimens, obtained from three species of terrestrial molluscs in Ontario, there is no overlap in ranges for total length, esophageal length, position of the nerve ring, position of the excretory pore, or length of spicules and gubernaculum (Table 2). Thus worms from molluscs were smaller in theserespects than worms from Amphibia. In these two seriesof measurements, there is some overlap in the size ranges of the tail of the male, the distance of the vulva from the anterior end, and in the width of both males and females. Without consider ing additional criteria, these two groups of nematodes appear different. However, when the ratios of the length of the esophagus, the length of the tail, the position of the excretory pore, the position of the nerve ring, and the position of the vulva are compared in relation to total body length, there is overlap of all ranges (Table 3).
Thus, the differences first noted, involving size alone, lose their apparent significance. A comparison of these 36
TABLE 2
MEASUREMENTS (IN MM) OP COSMOCERCOIDES DUKAE PROM ONE AMERICAN TOAD (BUFO AMERICANUS)PROM OHIO AND C. DUKAE REPORTED BY ANDERSON (I960) PROM TERRESTRIAL MOLLUSCS IN ONTARIO
Ohio H o st: Ontario H o sts: One American toad M olluscs—■3 species
Length, male 3.251 - 4.725 1.65 — 2.98 Length, female 3.888 - 5.905 1.66 - 4.34
Width, male 0.179 _ 0.252 0.12 — 0.30 Width, female 0.179 _ 0.278 0.15 - 0.31
Esophagus, male 0.730 _ 0.863 0.35 — 0.38 Esophagus, female 0.809 - 1.022 0.33 - 0.52
Nerve ring, male 0.292 0.360 0.16 — 0.28 Nerve ring, female 0.285 - 0.350 0.14 0.27
Excretory pore, male 0.431 — 0.530 0.24 _ 0.38 Excretory pore, female 0.471 — 0.597 0.18 — 0.42
Spicules 0.373 - 0.478 0.20 - 0.32
Guvernaculum 0.129 - 0.163 0.07 - 0.11 Vulva, to anterior 2.176 - 3.304 0. 88 - 2.42
T a il, male 0.152 0.189 0.11 _ 0.17 Tail, female 0.205 0.305 0.12 0.19
No. of nematodes male 24 20 female 24 10 37
TABLE 3
DERIVED STATISTICS (PERCENTAGES) OP C. DUKAE PROM ONE AMERICAN TOAD PROM OHIO AND C. DUKAE REPORTED BY ANDERSON $1960) FROM TERRESTRIAL MOLLUSCS IN ONTARIO
Ohio Host: Ontario Hosts: One American toad Molluscs—3 species
Esophagus/body len g th x 100 male 18 - 24 (22) % 13-21 (17) % female 17 - 22 (20) % 10 - 19 (15) %
Nerve ring/ body length x 100 male 6 - 10 (9) % 1-12 (9) % female 6 - 8 (7) % 5 - 8 (7) %
Excretory pore/body length x 100 male 11 - 15 (14) % 10-15 (13) % female 10 - 13 (12) % 8-12 (10) %
Tail/body length x 100 male 4-5 (4.8) % 4 - 8 (6) % female 5-6 (5.4) ^ 4 - 8 (7) %
Vulva, to anterior/body length x 100 55 - 58 (57) % 53-57 (55) % No. of nematodes male 24 10 female 24 10 38 two groups of worms with respect to cephalic papillae, shape of the reproductive system, spicules, and guber- nacula reveals close agreement. Specimens of C. dukae from gastropods were loaned by Dr. R. C. Anderson of the
Ontario Research Foundation. An examination of these revealed no striking morphological differences between them and specimens taken from Ohio amphibians.
In order to evaluate size variations among C. dukae collected in Ohio amphibians, three groups of worms have been described statistically (Tables 4 through 15). The first group consists of 48 specimens from one American toad col lected from Laurel Township in Hocking County (Tables 4 through 7). The second group consists of the 48 specimens of group one added to 25 specimens from twelve other species of
Ohio amphibians (Tables 8 through 11). The third group con sists of eleven worms taken from five host species in the same lo c a tio n as group one, but not from the same in d iv id u a l host (Tables 12 through 15). With respect to hosts, the second group consists of specimens from the ten infected host species listed in Table 1 in addition to Plethodon g lu tin o su s, Ambystoma opacum, and Pseudacris t r i s e r i a t a .
The hosts in the third group were Desmognathus fuscus,
Eurycea bislineata, Rana sylvatica, Pseudacris brachyphona, and Bufo americanus. Data for males and for demales of each group are listed both as direct measurements and as derived TABLE 4. Mil© m easurements, Group I ; ^8 specimens from one Ameri ,... (in mm.) from Laurel Twp., Hocking C WORM LENGTH WIDTH ESOPHAGUS NERVE RI1 NUMB ER 5 1 3.8»*6C 0. 1990 0 . 7360 0.3*400 52 5 .2 5 1C 0. I860 0. 7**20 0.2980 5? *4 . 725C 0.2520 0.8630 0.3050 5** 3. **0 9 0 0. 1990 0.8C90 0.33*40 55 *4.07*4C 0.2390 0.8360 0.2920 56 3 . 39 7 C C. 1990 0.7700 0.3390 57 3.75 5C 0.1970 0.7560 0.3280 58 3.5*+3C 0. 1 990 0.82 30 0.3260 59 3.29 1C C. I860 0.7500 0.298C 60 3.675C 0. 1990 0.7630 0.32UC 6 1 3.90 1C 0.2 120 0.7960 0. 323C 62 3.149CC 0.2 120 0.7830 0.336C 63 3. 7**2 0 0.2050 0.7960 0.33 It 6** 3.596C 0.209C 0.7960 0. 32** C 65 3 . ** 50 C 0.1790 0.71*30 0.305C 66 3.8220 0.20 5 0 0. 7 9 60 0. 33**C 67 3.39 7 0 0.2050 0 .756u 0.333C 6d 3.556C 0.2 T2C 0.8C90 0 . 32*+C 6V 3.53CC 0.2 120 0.7** 30 0 .339C 70 3 . 5 1 6 C 0.2 26 0 0.7960 0. 3 6 0 ( 7 1 3 . ** 6** C 0.1920 0.7 6 30 0 . 3 1 5 C 72 3 . 3 ** ** C 0.2060 0.7560 0. 3 1 7( 15 3 .6630 0. 1990 0.7700 0. 3 I2( 7*4 3 . ** 9C C 0.206C 0. 7**30 0. 325(
VARI ANCE 0 . C V 61* 0.00 0 3 0.0C 12 0.000.
MEAN 3.622C 0.2036 0.7797 0. 3 23*
ST ANO ARC C E V I A T I C N 0 . 3 1 C 5 0.0 160 0.0 3*4 1 0.0 1 61
ST AN CARD ERRCR 0 .L63** 0.00 3 3 0.0C70 0.003
RANGE H ICH **. 7250 C.2520 0.8630 0. 3 6Ci
LCW 3.25 1C 0 . 1 7 9 C 0.7300 0.292 (V O' <3« *H •H O 0*0 a O K O C D O CD £ hO CD o o cd c
C toad (Bufo am ericanus) 4-3 cx >3 O
< CO ss w PS E-i £> (a Pi s, •< 1—1 •< o § £ 5z E-* CL, g CO O, g CO o w a M o — ——«—— o nO o o o o o o o o CMro o o o o o 3 X o o o o — 3 o p- 3 o _ o o o o o o o l >o 3 O f • • • • 4 o i/l CM CM o 3 N- - w •> o o -0 3 o 3 o o o —■ o o 3 3 3 o • • • 4 • o o "U ro o o x o o no o ro o un o *— —■ — o o -oooX o o o p- X o o • • • • • • o o o o o o p- 0CO-0 > ro o ro o o o m o o o p- o o o 3 3 -3* • f • 4 « • • o "0 o CM o o o CM CM o — no o o OOO o o CM ■o a • 4 o o o p* o o ro ro ro o CM o o o o o i/1 o o — p- 3 o 3 • • • • • • 4 O -p-p- X o — — 3 o o CMCMX o o ooo o Oo o o 3 in o CM C->o o- nO -3 ro Jf -3- • • rO o O o MX> X CM ro o o ro ooo o Oo o O
ii 00 0.2920 0.4310 0.3730 0.1290 0.1520 40
TABLE 5. Female measurements, Group I; 48 specimens from one American To< WCRf* (in mm.) from Laurel Twp., Hocking County, NUMBER LENGTH WIDTH ESOPHAGUS NERVE RING EXCRE1 75 *4 . 68 4 C 0. I99C 0.9C2C 0.2850 C . 4' 7 6 3.88 8C 0.1920 0.8*490 0.3120 C. 4i 7 7 4.233C 0.2 190 0.8 6 30 0.3260 C.5i 78 4. 10 1C 0.1920 0.8630 0.3200 0.4 7 9 *4 .03*40 0.1790 0.8 360 0.3120 C • 4' 80 U. 05*4 0 0. 1920 0.8630 0.3 360 0. 5i 8 1 *4.592 0 0.1920 0.8*490 0.3170 0.5 82 *4. I 1 *4 C 0. 192 0 0.8490 C . 3180 0.4 83 *4. 1880 0. I860 0.8230 0.3250 0.5 8*4 4.4 19C 0.2050 0.8890 0.3 380 0.5 85 5.52CC 0.278C 1.0220 0.3380 0.5 86 *4. 5660 0. 1990 0.8760 0.3 180 C. 4 8? *4 . 26C C 0. 1990 0.86 30 C.3 120 C. 5 88 *4 . 5650 0.2190 0.9420 C.3380 0.5 89 *4. C 6 C C 0. 1 79C 0.8230 0.320C 0.4 9 C *4. 68*40 0.225C 0.9420 0.3420 C . 5 9 1 *4 . 5650 0. 1 990 0.9290 u . 3 4 5 C 0.5 92 *4. 10 c C C.189C 0.8490 0.3220 0.5 9 5 *4.06 1C 0. 1 79 0 0.8C9C C.3120 0.4 9*4 *4. 1950 C.2250 0.849C C.3080 0.4 95 5.888C 0. 1920 0.8C90 0.3250 0.4 96 *4.6 180 0.209C 0 .9 I6C 0.3 5 20 0.5 97 *4 . 2*4 7 0 0.1920 0.8 7 60 0.3 320 0.4 o o 9 8 5.9050 0 . 2 7 8 C 1 .0220 • C. 5
VARIANCE 0.2255 0.0CC7 0 .00 33 C . 0 C C 2 0.0
N E AN *4 . 37 16 0 . 2 C *4 6 0.8797 C . 3 2 4 3 0.5
ST ANDARC CfcV 1 AT IGN 0 . *4 7 *4 6 0.0262 0.0578 0.0145 O.C
STANDARD ERKCR 0.096V 0.005*4 0.0118 C . 0 0.30 O.C RANGE 8 I GH 5 .90 5 0. 0.2 78C 1 .0220 0.3510 o.c
LCW 5 . 8P8C L . 1 7 9 C 0 . 8 >_ 9 0 0.2850 C.* ■ican Toad, Bufo americanus. C o u n ty , O h io . VULVA VULVA EGG EGG ING EXCRET.PORE TAIL TO ANTERIOR TO POSTE-f IOR LENGTH 'tflDTH ) C.4970 0.2 110 2.7070 1 .9770 0 .0820 0.0350 ) C.4840 0.2260 2.2030 1 .6850 0.08 10 0.0400 ) C.504C 0.2520 2.3350 1.8980 0.0830 0.0450 l o.uouc 0.2 120 2.32 30 1.7780 0.0770 0.0420 } C.4970 0.2 120 2.3090 1.7250 0.078G 0.0420 ) 0.5040 0.2 190 2. 31+90 1.6850 0.0780 0.0430 5 0.524C 0.2450 2.5080 1 .881+0 0.071+0 0.0380 ) 0.1+01+0 0.2220 2.3890 1.7250 0.0770 0.0430 ) 0.5040 0.2C50 2.1+230 1.7650 0.0700 0.0 3 70 ) 0.5 300 0.2390 2.5U 80 1.87 10 0.0800 0.0430 ) 0.58 30 0.2980 3. 10 50 2.1+ 150 0 .08CG 0.0460 3 C.4970 0.2250 2.1+680 1 .8980 0.C800 0.04 30 J C.5 1 10 0.2590 2.1+020 ■ 1 .8 580 C.08 50 0.0420 3 0.5300 0.2590 2.57U0 1 .99 1C 0.0850 0.04ou ; 0.1+81+c 0.2320 2.2690 1 .79 10 0.0 7 70 0.0400 3 C.5900 0.2240 2.6800 2.CC1+0 0.09 30 0.04 10 j 0.5300 0.2320 2.1+810 I.884C 0.C900 0.0370 3 0.517C 0.2 320 2.28 20 1 .8 180 0.0720 0.0420 3 0.1+710 0.2 190 2.3090 1 . 7520 0.0 720 0.0430 3 0.1+81+0 0.2390 2.3880 1 .8050 0.0800 0.0450 3 O.i+Bi+O 0.2 190 2. 1 760 1 .7 120 0.C800 0.0380 0 0 . 5' 3 7 C 0.2520 2.571+0 2.044C 0.0 7 70 0.0420 3 0.1+910 0.2 190 2. 349u 1 .8980 0.0780 0.04 30 0 C.5970 0.3 05 0 3. 30U0 2.60 10 0.0850 0.0450
2 0.0012 0.0006 0.0696 0.01+68 o.cooc 0.0000
3 0.5 132 0.2 357 2.H77 3 1 .891+3 0.0797 0.04 1 7
5 0.0 31+7 0.0253 0.2639 0.2 163 C . 0 0 5 1+ 0 . 0 C 5 u
0 0.0071 0.0 C 52 0.C5 39 0.01+1+2 0.00 1 1 0.0006
0 0.59 70 0 . 3C50 3. 301+0 2.6010 0.0930 0.0460
0 C.1+7 10 0 . 2 C 5 0 2. 17 60 1 .6850 C.07CG 0.0350 41
TABLE 6. Jfale derived statistics, Group I: 48 specimens from one America (mm.) from Laurel Twp., Hocking Cou ESOPHAGUS EXCRET.PORE NERVE RING TAIL BODY WORM BODY LENGTH BODY LENGTH BODY LENGTH BODY LENGTH *.rr* NUMBER X 100 X 100 X 100 5 1 19. 1268 13.4356 8.8358 4.3 139 19. 52 22.823 7 14.2725 9.1664 4.6755 17. 53 18.2645 I 1.2 169 6.4550 3.9365 18. 54 23.7313 13.9924 9.7976 4.8695 17. 55 20.5204 10.5793 7.1674 4.1237 17. 56 22.667 1 13.8652 9.9794 4.8867 17. 57 20. 1332 13.9547 8.7350 4.6 338 19. 58 23.2289 13.8301 9.20 12 4.7982 17. 59 22. 10 1 7 13.7040 9.0550 5. 1960 17. 60 2 1 .306 1 13.5238 8.8 163 4.9 5 24 18. 6 1 20 .40 5 C 13.0736 8.2 799 4.4604 18. 62 22.4355 14.6132 9.6275 5.2149 16. 63 2 1 .2720 1 3.4687 0.8455 4.9 103 18. 64 22 . 1357 13.2647 9.0 100 5.0 3 34 17. 65 2 1 .5362 I4.4C58 8.8406 4.695 7 19. 6 6 20.8268 13. 1868 8.7389 4.6049 18. 67 22 .2549 13.659 1 9.0C28 4.8867 16. 68 22.7503 13.8076 9.1 114 5.0056 16. 69 2 1 .0482 14.2776 9.6C34 5.354 1 16. 70 22.6394 14.70 42 10.2389 5.3185 15. 7 1 22.6039 13.77C2 9.0935 4.9942 18. 72 22.6077 14.4737 9.4797 5.26 32 16. 73 21.02 1C 1 3.4C43 8.5 176 4.8594 18. 7 4 21.2894 14.2407 9.3 123 5.1003 16.
VAR I ANCE 1.7 184 0.9095 0.6863 0.1309 1 .
RE AN 2 1.6171 13.6 135 8.988C 4.8328 17.
STANDARD DEVIATION 1.3 109 0.9537 0.8284 0.3619 1. STANDARD ERRCR 0 . 2 6 7 6 0. 1947 0. 1 69 1 0.0 7 39 0. RANGE HIGH 2 3.7.-512 14.7042 10.2389 5.3541 19
LOW 18.2645 10.5793 6.4 5 50 3.9365 1 5 ’ican Toad, Bufo americanus C ou n ty, Ohio. 30DY LENGTH BODY LENGTH BODY LENGTH SPICULE Y'lDTH ESOPHAGUS TAIL GTJHERNAClL/M
19.3367 5.2283 23. 1807 2.79 19 17.4 7 85 4.38 14 2 1 .3882 2.8299 18.7500 5.475 I 25.4032 2.98 7 5 17.1307 4.2 138 20.536 I 2.5478 I 7.0460 4.8732 24.2500 3.0388 I 7.0704 4.4 117 20.4639 2.63 16 19.0609 4.9669 2 1 .5805 2.7234 17.8040 4.3050 20 .84 12 2.687 I 17.6935 4.5082 19.2456 2.5578 18.4673 4.6935 20. 1923 2.5 190 18.4009 4.9008 22.4 195 3.0000 16.4623 4.4572 19.1758 2.5772 18.2537 4.70 10 20.7889 2.7397 1 7.2057 4.5 176 19.8674 2.5 185 I 9 . 2 7 3 7 4.6433 21.2963 3.2090 18.64 39 4.8C 15 21.7159 3.1973 16.5707 4.4934 20.4639 2.9C9 1 16.7736 4.3956 19.9775 2.7945 16.6509 4.75 10 18.6772 2.5 1 32 15.5575 4.4 17 I 18.802 I 2.5287 18.04 17 4.4240 20.023 1 2.5 185 16.2330 4.4233 19.0000 2.5C67 18.4070 4 .757 1 20.5787 2.6835 16.9417 4.6972 19.6067 2.5903
1.0626 0.0 806 2.8044 0.0488
17.6356 4.64 32 20.8 I 14 2.7534
1.0308 0.2976 1 .6746 0.22 10
0.2104 0.0 60 8 0.34 18 0.045 1
19.3367 5.475 1 25.4032 3.2090
15.5575 4.2 1 58 18.6772 2 . 5 C 6 7 42 TABLE 7. Female derived statistics, Group I: 48 specimens from one Arne (in mm.) from Laurel Twp., Hocking ESOPHAGUS EXCRET.PORE NERVE RTNG TAIL BO WORN ~8GOY LENGTH b50Y M O W UfffiY 'LENGTH BODY LENGTH ~ NUMBER X 100 X 100 X 100 x 1 0 0 75 19.2570 10.6 106 6.0845 4.5047 2 76 2 I .8 564 12.4486 8.0247 5.8128 2 77 20.3874 1 1 .9064 7.70 14 5.9532 1 78 2 I .0436 1 1.8020 7.8030 5.1695 2 79 20.7238 12.3203 7.7343 5.2553 2 80 2 1.3932 12.4938 8.3292 5.4289 2 8 1 I9.33C6 1 1 .9308 7.2 177 5.5783 2 82 20.6368 1 1 . 7647 7.7297 5.3962 2 85 19.65 14 12.0344 7.7603 4.8 94 9 2 84 20. 1 177 1 1 .9937 7.6488 5.4085 2 85 18.5 145 10.56 16 6. 1232 5.3986 1 86 20.064 1 11.3834 7.2836 5.1535 2 87 20.2582 11.9953 7.3239 6.0798 2 88 20.6353 11.6101 7.4042 5.6736 2 89 20 .2709 1 1.92 12 7.88 18 5.7 143 2 90 20 . 1 1 1C 12.596 1 7.3C 15 4.7822 2 9 1 2 1 .2829 12.1420 7.9038 5.3150 ' 2 92 20 . 70 73 12.6098 7.8537 5.6585 2 9 8 19.92 12 1 1 .598 1 7.6828 5.3928 2 94 20 .2480 11.5430 7.3456 5.7000 1 95 20 .8076 12.4486 8.3591 5.6327 2 96 19.8354 1 1.6284 7.1893 5.4569 2 97 20 .6263 1 1 .56 11 7.8 173 5.1566 2 98 17.3074 10. 1 10 1 5.944 1 5.165 1 2
VAR I ANCE 0 .9084 0.4078 0.40 14 0.1328
N E AN 20.20 70 11.7922 7 .4 7 70 5.4034 2
STANDARC DEVIATION 0.953 1 0.6386 0.6336 0.3644
STANDARC ERROR 0.1945 0. 1304 0.1293 0.0744 RANGE h I GH 2 1 .8364 12.609 8 8.359 1 6.0798 2
LOk. 17.3074 10. 1 1C 1 5.944 1 4.5047 1 ,e American Toad, B'ufo ’ americanus , icking County, Ohio. VULVA VULVA BODY LENGTH BODY LENGTH BODY LENGTH TO ANTERIOR TO ANTERIOR ST h Width "e S'OMAGuS fAIL BODY LENGTH BODY LENGTH X 100 7 23.5377 5. 1929 22. 1991 0.5779 57.7925 8 20.2500 4.5795 17.2035 0.5666 56.66 15 2 19.3288 4.9050 16.7976 0.5516 55. 16 18 5 2 1. 3594 4.7520 19.3443 0.5664 56.6447 3 22.5363 4.8254 19.0283 0.5724 57.2385 9 21.0104 4.6744 18.420 1 0.5823 58.2300 3 22.8750 5. 1731 17.9265 0.57 10 57. 1038 2 21.4271 4.8457 18.53 15 0.5807 58.0700 9 22.5 16 1 5.0887 20.4293 0.5786 57.8558 5 2 1.556 1 4.9708 18.4895 0.5766 57.660 1 6 19.8561 5.4C 12 18.5235 0.5625 56.2500 5 2 1 .9397 4.9840 19.4044 0.5653 56.5277 8 2 1 .4070 4.9363 16.4479 0.5638 56.3850 6 2C.8447 4.846 1 17.6255 0.5639 56.3855 3 22.68 16 4.9332 17.5000 0.5589 55.8867 2 20.8178 4.9724 20.9 107 0.5722 57.2161 o • 2 1 .9347 4.6986 18.8 147 0.5684 56.8385 5 2 1.693 1 4.8292 17.6724 0.5566 55.6585 8 22.6872 5.0 198 18.5434 0.5686 56.8579 0 18.6356 4.9388 1 7.5439 0.5695 56.9521 7 20.2500 4.8059 17.7534 0.5597 55.9671 9 2 2.0957 5.04 15 18.3254 0.5574 55.7384 6 22. 1 198 4.8482 19.3927 0.553 1 55.3096 1 2 1 .24 1C 5.7 7 79 19.3607 0.5595 55.9526
8 1 . 392 1 0.0622 1 .7193 0.0C0 1 0.7388
4 2 1.4417 4.9600 18.59 12 0.5668 56.68 10
4 1. 1799 0.2494 1.3112 0.0086 0.8596
4 0.2408 0 .050 9 0.2677 0.00 18 0.1755
3 2 3.53 7 7 5.7779 22. 199 1 0.5823 58.2300
7 18.6356 4.5795 16.4479 0.55 16 55. 16 18 43 TABLE 8. Male measurements, Group II : 48 specimens from one American WORM (in mm.) from 12 other species of Ohio NUMBER LENGTH IDTH ESOPHAGUS NERVE RING' EXC 3ft 2.495C 0. 1720 0.4380 0.2000 0 37 4.6 18C 0.259C 0.66 30 0.2650 0 38 2.057C 0. 1460 0.3980 0. 1 7 1C 0 39 1 - 9 7 7 C 0.1720 0.4370 0. 1980 0 40 2.9050 0. I860 0.4570 0.2080 0 4 1 3.38fc 0 0. 1 720 0.5600 0. 1840 0 42 3.688C 0.2720 0.6560 0.2540 0 4 3 3 . C 9 2 C 0. 1 86 C 0.5500 0.2480 0 44 2.4950 0. 1600 0.4970 0.1620 0 45 4.80CC 0.4520 0.8C2C 0.2980 0 46 5 .2960 0.4310 1 .00 10 0.3450 0 4 7 4 . 7 2 C C 0 .398C 0.95 10 0.37 10 0 48 5.055C 0.3780 0.8 100 0.2670 0 49 3 . 0 7 1 C 0 .329C 0.6590 0.2450 0 50 2 .8260 0.2050 0.4540 0.24 10 0 5 1 3 . 84 8 C 0. 1 9 9 C 0.7360 0.3400 0 52 3.25 10 0. I860 0.7420 0.2980 c 53 4.7250 0.2520 0.8630 0.3050 0 54 3 . 4 C 9 C 0. 1990 0.8C90 0.3340 0 5 5 4 .0740 0.239C 0.8360 0.2920 c 56 3.3970 0. 1990 0.770C 0.3390 c 5 7 3 . 755.. 0 . 1 9 7 0 0.7560 0.3280 0 58 3.5430 0. 1990' 0.8230 0. 3260 0 59 - 3.29 I C 0 . I860 0.7 300 0.2980 0 60 3.6750 0. 1990 0.7830 0.3240 c o 1 3 .90 1 C C.2 120 0.7 96 0 0.32 30 0 -62 3.4900 0.2 120 0.7830 0.3 360 c 63 3.74 2 0 0 .2050 0.7 9 60 0.33 10 0 64 3.59 6 0 0.2U9C 0.7960 0.3240 c 6 5 3 . 4 5 C 0 0.1790 0.7430 0.3050 0 66 3.8220 0.2050 0.7960 0.3 340 c 67 3.39 7 0 0.2050 0.7 560 0.3 3 30 0 68 3.5560 0.2 1 20 0.8C9G 0.3240 0 6 9 3.5 30 0 0.2 120 0. 7430 0.3390 0 70 3.5 160 0.226 0 0.7960 0.3600 0 7 1 3 .46 4 C 0. 1920 0.7830 0.3 150 0 72 3.34 4 0 C.206C 0.7560 0.3 170 0 7 3 3.6630 0. 1 99 C 0. 7 700 0.3 120 0 74 3.4900 0.206 0 0.7430 0.3250 0
VARIANCE 0.52 7 9 C . 0 0 5 2 0.0205 0.0050 c
MEAN 3.5746 0.2270 0.7 192 0.2928 0
STAUCARD DEVIATION 0 . 7265 0.0723 0.1431 0.0552 c
ST ANUARD ERRC R 0.1163 0.0 116 0.0229 0.0088 0 RANGE HIGH 5.2960 C.4520 1 . C C 1 c 0.37 10 0
LOW 1 .9770 0. 1460 0.3980 0. 1620 0 ?ican Toad added to 25 specimens Ohio Amphibians. 7G-EXCRET.PORE SPICULES GTTBEENACUL'TM TAIL 0.2980 0.2670 0.1150 0. 1360 0.4640 0.3 120 0.13 10 0.17 10 0.2890 0.2300 0.0930 0.1360 0.2890 0.2240 0.0920 0. 1460 0.3360 0.2300 0.0900 0. 1660 0.3980 0.2240 0.0940 0. 10 40 0.3980 0.2880 0 . 1120 0. 1390 0.3780 0.3040 0.14 10 0. 1600 0.3650 0.2640 0. 1000 0. 1680 0.4840 0.4C80 0. 1520 0. 1920 0.5700 0.4640 0. 1660 0.2090 0.5200 0.4770 0. 1690 0. 1990 0.4 440 0.3800 0. 1400 0. 1840 0.3820 0.3340 0 . 1020 0. 1400 0.3440 0.2850 0. I 180 0.1280 0.5 I 70 0.4 160 0. 1490 0. 1660 C. 4640 0.4 160 0. 1470 0. 1520 0.5300 0.4780 0. 1600 0.1860 0.4770 0.4000 0. 1570 0. 1660 0 . 4 3 IC 0.3920 0. 1290 0. 1680 C.47 10 0.4C00 0. 1520 0. 1660 0.5240 0.384C 0.14 10 0. 1 740 0.4900 0.4380 0. 1630 0. 1700 0.45 1C 0.3760 0. 1470 0.17 10 C.4970 0.3980 0. 1580 0. 1820 0. 5100 0.4C80 0. 1360 0. 1 740 C.5 I00 0.3840 0. 1490 0.1820 0.5040 0.4C00 0. 1460 0.1800 C . 4 7 7 0 0.4C80 0. 1620 0.18 10 0.4970 0.4300 0. 1340 0. 1620 C.504C 0.4700 0. 1470 0. I 760 0.4640 0.4 4 80 0. 1540 0. 1660 0.49 10 0.4C80 0.1460 0. 1780 0.5040 0.3820 0.1520 0. 1890 > 0.5170 0.3970 0. 1570 0. 1870 0.4 770 0.4C80 0. 1620 0. 1730 0.4840 0.3 760 0.1500 0. I 760 0 . 4 9 I C* 0.4 240 0. 1580 0. I 780 0.4970 0.3730 0. 1440 0. 1 780
C.005 I 0.0053 0.0005 0.0004
0. 454H 0.37 19 0.1388 0.1682
C.07 15 0.0 7 30 0.C230 0.0206
0.0115 0.0 117 0.00 3 7 0.0033
0.5700 0.4 780 0.1690 0.2090
0.2890 0.2240 0.0900 0 . 1C40 44
TABLE 9. Female measurements, Group II: 48 specimens from one American Tc *ORM' (in mm.) from 12 other species of Ohio amp NUMBER LENGTH WIDTH ESOPHAGUS NERVE RING EXC 75 4 • 68 4C 0 . 1 9 90 0.9C20 0.2850 0 76 3.888C 0.1920 0.8490 0.3 120 0 77 4.2330 0.2 190 0.8630 0.3260 0 78 4. 10 1C 0. 1920 0.8630 0.3200 0 79 4.034C 0.1790 0.8360 0.3 120 0 80 4.C34C 0. 1920 0.8630 0.3360 0 8 1 4.392C 0.1920 0.8490 0.3170 0 82 4.1140 0. 1920 0.8490 0.3180 0 8 3 4. 188C 0. I860 0.8230 0.3250 0. 84 4 .4 19C 0.2050 0.8890 0.3300 0 85 5.52CC 0.2780 1.0220 0.3380 0 86 4.3660 0. 1990 0.876C 0.3 180 0 87 4.26GC 0.1990 0.8630 0.3 120 0 88 4.565C .0.2 190 0.9420 0.3380 0 89 4.C60C 0. 1790 0.8230 0.3200 0 90 4 . 6 8 4 C 0.2250 0.9420 0.3420 c, 9 1 4.365C 0. 1990 0.9290 0.3450 0. 92 4 . 1C C C 0.1890 0.8490 0.3220 0. 9 5 4.06 10 0.17 90 0.00 90 0.3120 0. 94 4. I93C 0.2250 0.8490 0.3080 0, 95 3.888C 0. 1 920 0.8C90 0.3250 0. 96 4.6 180 0.209C • 0.9160 0.3320 0, 97 4 .24 7C 0 . 19 20 0.876C 0.3 320 c, 9 8 5.905C 0.2780 1 .0220 0.35 10 0. 99 5.3740 0.4 180 0.6570 0.2850 0, 100 3 . 1 9 8 0 0. 1 57 0 0.4300 0.1920 0. 10 1 3.5750 0.26 5 0 0.4 5.7 G 0.2000 c, 102 4.3660 0.2650 0.5440 0.2290 0. 105 4.9760 0. 1650 0.53 10 0.2 180 0. 104 2.9 1 9 C 0.1330 0.5 700 0.2290 0. 105 6.1170 0.3720 0.6 900 0.2650 0. 106 5. 75 1 C C.3780 1 .0 I4C C.3380 0. 107 5.269C 0.4CCC 0.7480 0.2 760 0. 100 5. 100 0 0.2 750 0.6230 0.24 80 0.
VARIANCE 0.5065 0.0049 0.0238 0.0019 0.
MEAN 4.4578 0.22 7 6 0.8049 0.30 19 0.
ST ANDARC CEV I AT ION C.7II5 0.069 8 0.1544 0.0439 c.
STANDARC ERROR 0 . 122 0 0.0 1.20 0.0265 0.0075 c. RANGE HIGH 6.1170 0 . 4 1 8 C 1 .0220 C . 3 5 1 0 c.
LCk 2 . V 1 9 C 0 . 15 5 0 1 0 . 4 5 8 C 0. 1920 0.
l 0 0 0 0 c 0 0 o o o o O o o o 0 o o o o O o O o o o o o o O o o a o o CDO o o o • •• • •• •• •• • • • • • •• •• •••• •• • • 0 • • •• • • •• 0 • • • • IO l t o o j r o Lm • p LT LM ro LM Ua LM tr LT •P Ln tr XT tr LH Ln i n P U 1 l / l P i / i l / l l / l p l / l l / l p p l / l p P a
— ro o o — O ?o ro ro ro _ IO ro ——- ro ro ro _ —_ _ —ro —_ ro • • • • • • • • •• • •• • • ••• • • • • • •• • • • • • • •• • • ••• • • — -0 o Lm O — f ■tr Ln-g ro o O' tr Ln a cc o -si o: -gCDCDO -gO 0D00 tr OD-g -gCD O -g-g0Do o —- -sj o Ln o ro o Lm Lm-si —CD ln ro Lmo t - o Ln CDo <5>cLn -o -g o ro CDft ro -gC CD-g Ln LM o tr ro LP LmO .r LMLnro -Sl 0DLmO —CDtr ro Ln ro CDtr tr 0DODlt —LnLn tr Ln LT CDCDLP -g o o -g — Lm tr O O o o o o o O O O o o O o o o O o o o o o o o o o o O o o O O o o a 0 0 0 0 0 0 0 0 0 0 o o o o o O o o o o o o o CDo o o O o o o o o o CDo CDCDo 0 •• • • • •••• •• • 0 • • • • • •• • ••• a • • • • • » • • • • • ■ • • • tc o o o o o o o o o o c o o o o o CDo o CDo o o CDO O o CDo CDCDo o o o CDO o o o 5*3 G M tr o -sj o -go cr l/l O' p p O' CD-sj CDCD-o -si sO '0 —1 00 00 03 00 CO-J —1—I -J -J -J 30 00 CD Q Q tr Lm ro tr o o o O CD O' M ro p p p l/i OD^1 CDo ro ro o Ul-4 l/l 1/ CDo o o -»Jp 0303 Ui _ ro •-3 O OC‘ Ln Lm — o o o o o O o Cj o o O O o O c o c o O CD o o O o o CD o o o O o O a o W 0 0 0 0 0 0
o o 0 o o o o o o o o o o G OO o o O o o o O 0 o o o O o o o a OO • • • • • • • • • • • ■ • • • •• • • • • • • • • • • • • • • • t 0 0 - 0 0 0 0 o o o o o o o o o o o o o o O o o o o o o o o o o o o © G o o o o o o o o o o o Nj r - o o c . r - tr p u. p p p ro \3 u p p p Uj p p p Ui tr p PP p P Uip u p p p p p u LT a o LT o o tr u c. o l/l O' a- I/' I/. u ro OD l/l u ro —I _ O O' ro U O' U -J u a u ro ro IT G 1/ C> o IO o o c a c o o G o G O G o G G O o c G G OO G O GG o O G o G o G G G G TABLE 10 • Male derived s t a t i s t i c s , Group II; 48 specimens from one Amer: ( i n m m .) from 12 other species of 0] ESOPHAGUS EXCRET.PORE NERVE RING TAIL.. BODY LE WORM BODY LENGTH BODY LENGTH NUMBER BODY LENGTH BODY LENGTH WIDT X 100 X 100 X 100 36 17.555 1 1 1.9439 8.0 160 5.4509 14.50 37 14.3569 10.0476 5.7384 3.7029 17.83 38 19.3486 14.0496 8.3 131 6.6 1 16 14.08 39 22. 1042 14.6 18 1 10.0 152 7.3849 1 1.49 40 15.7315 1 1.5663 7.1601 5.7 143 15.6 1 4 1 16.5387 1 1. 7543 5.434 1 3.07 15 19.68 *42 17.7874 10.79 18 6.8872 3.7690 13.55 43 17.7878 12.2251 8.0207 5.1746 16.62 44 19.9 198 14.6293 6.4930 6.7335 15.59 45 16.7083 10.0833 6.2083 4.0000 10.6 1 46 18.90 1 1 10.7628 6.5 144 3.9464 12.28 47 20. 1483 1 1.0 169 7.8602 4.2161 1 1 .85 48 16.0237 8.7834 5.28 19 3.6400 13.37 49 21.4588 12.4389 7.9779 4.5588 9.33 50 16.065 1 12. 1727 8.5280 4.5294 13.78 5 1 19. 1268 13.4356 8.8358 4.3139 19.33 52 22.8237 14.2725 9.1664 4.6755 17.47 53 18.2645 1 1.2169 6.4550 3.9365 18.75 54 23.73 13 13.9924 9.7976 4.8695 17. 13 55 20.5204 10.5793 7.16 74 4.1237 17.04 56 22.667 1 13.8652 9.9794 4.8867 17.07 57 20. 1332 13.9547 8.7350 4.6338 19.06 58 23.2289 13.830 1 9.20 12 4.7982 17.80 59 22. lb 17 13.7040 • 9.0550 5.1960 17.69 60 2 1 .306 1 1 3. 5238 8.8 163 4.9524 18.46 6 1 20.4C5C 1 3.0736 8.2799 4.4604 18.40 62 22.4355 14.6 132 9.6275 5.2 149 16.46 63 2 1 .2720 13.4687 8.8455 4.8 103 18.25 64 22. 135 7 13.2647 9.0 100 5.0334 17.20 6 5 2 1.5362 14.4058 8.8406 4.6957 19.27 66 20.0260 13. 1868 8.7389 4.6049 18.64 6 7 22.2549 1 3.659 1 9.8C28 4.8867 16.57 68 22.7503 1 3.8076 9.1114 5.0056 16.77 69 21.0482 14.2776 9.6034 5.354 1 16.65 70 22.6394 14.704 2 10.2389 5.3 185 15.55 7 1 22.6039 I3.77C2 9.0 935 4.9942 18.04 72 22.6077 14.4737 9.4797 5.2632 16.23 73 21.02 1C 13.4043 8.5 176 4.8594 18.40 74 2 1 .2894 14.2407 9.3 123 5. 100 3 16.94
VAR IANCE 6.0066 2.3992 1 .8052 0.6984 6.63
MEAN 20.237 1 12.9130 8.3 118 4.8 33 1 16.24
STANDARD CEVIATION 2.4508 1 .5490 1.3436 0.8357 2.57
STANDARC ERRCR 0.3924 0.2480 0.2 15 1 0.1338 0.4 1 RANGE HIGH 23.73 13 14.7042 10.2389 7.3849 19.68
LOW 14.3569 8.7834 5.28 19 £.0715 9.33 American Toad added to 25 s p e c i m e n s of Ohio am phibians. ODY LENGTH BODY LENGTH BODY LENGTH SPICULE WIDTH ESOPHAGUS TAIL GUBERNACULUM 14.5058 5.6963 18.3456 2.32 17 17.8301 6.9653 27.0058 2.3817 14.0890 5.1683 15.1250 2.473 1 11.4942 4.5240 13.54 1 1 2.4348 15.6 183 6.3567 17.5000 2.5556 19.6860 6.0464 32.5577 2.3830 13.5588 5.6220 26.5324 2.57 14 16.6237 5.62 18 19.3250 2. 1560 15.5937 5.0201 14.85 12 2.6400 10.6 195 5.9850 25.0000 2.6842 12.2877 5.2907 25.3397 2.7952 11.8593 4.9632 23.7 186 2.8225 13.3730 6.2407 27.4728 2.7 143 9.3343 4.660 1 21.9357 3.2745 13.7854 6.2247 22.078 1 2.4 153 19.3367 5.2283 23. 1807 2.79 19 17.4785 4.38 14 21.3882 2.8299 18.7500 5.4751 25.4032 2.9875 17.1307 4.2 138 20.536 1 2.5478 17.0460 4.8732 24.2500 3.0388 17.0704 4.4 117 20.4639 2.6316 19.0609 4.9669 2 1 .5805 2.7234 17.8040 4.3050 20.84 12 2.687 1 17.6935 4.5082 19.2456 2.5578 18.4673 4.6 935 20. 1923 2.5 190 18.4009 4.9008 22.4 195 3.0C00 16.4623 4.4572 19. 1758 2.5772 18.2537 4.70 10 20.7889 2.7397 17.2057 4.5 176 19.8674 2.5 185 19.2737 4.6433 2 1 .2963 3.2090 18.6439 4.8C15 2 1 .7159 3. 1973 16.5707 4.4934 20.4639 2.9C9 1 16.7736 4.3956 19.9775 2.7945 16.6509 4.75 10 18.6772 2.5 132 15.5575 4.4 17 1 18.802 1 2.5287 18.04 17 4.4240 20.0231 2.5 185 16.2330 4.4233 19.0000 2.5067 18.4070 4.757 1 20.5787 2.6835 16.94 17 4.6972 19.6067 2.5903
6.6374 0.463 1 13.1274 0.0621
16.2439 5.02 11 2 1.2770 2.6724
2.5763 0.6805 3.6232 0.2492
0.4 125 0 . 1C90 6.5802 0.0399
19.6860 6.9653 32.5577 3.2745
9.3343 4.2 138 13.54 I I 2. 1560 46
TABLE 11. Female derived statistics, G r o u p I I : 48 specim ens from one Am ( i n m m . ) from 12 other species of ESOPHAGUS EXCRET.PORE NERVE RING TAIL BOD WORM BODY LENGTH BODY LENGTH BObY LENGTH BODY LENGTH NUMBER X 100 X 100 X 100 X 100 75 19.25 70 10.6 106 6.0e45 4.5047 23 76 2 1 .8364 12.4486 8.0247 5.8 128 20 77 20.3874 11.9064 7.7C 14 5.9532 19 78 2 1.0436 I 1.8020 7.8C30 5.1695 2 1 79 20.7238 12.3203 7.7343 5.2553 22 80 2 1 .3932 12.4938 8.3292 5.4289 2 1 8 1 19.3306 1 1 .9308 7.2 177 5.5783 22 82 20.6368 I 1.7647 7.7297 5.3962 2 1 85 19.65 14 12.0344 7.7603 4.8949 22 84 20. 1 177 1 1.9937 7.6488 5.4085 2 1 85 18.5 145 10.56 16 6. 1232 5.3986 19 86 20.064 1 I 1 .3834 7.2836 5.1535 2 1 87 20.2582 1 1.9953 7.3239 6.0798 21 88 20.6353 1 1 .6 1C 1 7.4C42 5.6736 20 89 20.2709 1 1 .92 12 7.88 18 5.7 143 22 90 20. 1 1 10 12.596 1 7.3C 15 4.7822 20 9 1 2 1 .2829 12.1420 7.9038 5.3 150 2 I 92 20.7073 12.6098 7.8537 5.6505 2 1 93 19.92 12 1 1 .598 1 7.68 28 5.3928 22 94 20.2480 1 1.5430 7.3456 5.7000 18 95 20.8076 12.4486 8.359 1 5.6327 20 96 19.8354 1 1.6284 7. 1893 5.4569 22 97 20.6263 . 1 1 .56 1 1 7.8 173 5.1566 22, 98 17.3074 10.110 1 5.944 1 5. 165 1 2 1, 99 11.8534 7.8898 5.3033 3.9449 12, 100 13.696 1 9.9437 6.0038 5.3158 20, 10 1 12.7832 8.6993 5.5944 4.4755 13, 102 12.4599 8.1768 5.245 1 4.8557 16. 103 10.6712 5.7275 4.38 10 3.6977 30. 104 19.5272 12.0247 7.8452 5.5498 2 1, 105 1 1 .2800 8. 1249 4.3322 3.4658 16. 106 I 7.6317 9.1115 5.8772 4.260 1 15. 107 14. 1962 8.7872 5.2382 3.7958 13. 108 12.2 157 7.6078 4.8627 4.196 1 18.
VARIANCE 12.0829 3. 17 8 1 1.4 182 0.4593 1 1.
MEAN 18.2 730 10.856 1 6.8862 5.0953 20. srANCARD DEVIATION 3.476C 1 .7827 1. 1 909 0.6777 3.
STANDARC ERRCR 0.596 1 0.3057 0.2C42 0 . 1162 0. RANGE HIGH 2 1 .8 364 12.6098 8.3591 6.0798. 30.
LCW 10.67 12 5.7275 4^3 322 3.4658 12. American Toad added to 25 specimens of Ohio amphibians. VULVA ’ VULVA BODY LENGTH BODY 1ENGTH BODY LENGTH TO ANTERIOR TO ANTERIOR VfrDTH ESOPHAGUS TAIL BODY LENGTH BODY LENGTH X 100 23.5377 5.1929 22. 1991 0.5779 57.7925 20.2500 4.5795 17.2035 0.5666 56.66 15 19.3288 4.9050 16.7976 0.55 16 55. 16 18 2 1.3591+ 4.7520 19.3443 0.5664 56.6447 22.5363 4.8254 19.0283 0.5724 57.2385 21.0104 4.6744 18.4201 0.5823 58.2300 22.8750 5.1731 17.9265 0.57 10 57. 1038 2 1.4271 4.8457 18.5315 0.5807 58.0700 22.5161 5.0887 20.4293 0.5786 57.8558 2 1.556 I 4.9708 18.4895 0.5766 57.6601 19.856 I 5.40 12 18.5235 0.5625 56.2500 21.9397 4.9840 19.4044 0.5653 56.5277 21.4070 4.9363 16.4479 0.5638 56.3850 20.8447 4.846 I 17.6255 0.5639 56.3855 22.68 16 4.9332 17.5000 0.5589 55.8867 20.8 178 4.9724 20.9 107 0.5722 57.2161 21.9347 4.6986 18.8 147 0.5684 56.8385 2 1 .693 1 4.8292 17.6724 0.5566 55.6585 22.6872 5.0198 18.5434 0.5686 56.8579 18.6356 4.9388 17.5439 0.5695 56.9521 20.2500 4.8059 17.7534 0.5597 55.9671 22.0957 5.04 15 18.3254 0.5574 55.7384 22. 1 198 4.8482 19.3927 0.5531 55.3096 21.24 10 5.7779 19.3407 0.5595 55.9526 12.8565 8.4364 25.349 1 0.5285 52.8470 20.3694 7.30 14 18.8 I 18 0.54 13 54. 1276 1 3.4906 7.8228 22.343 7 0.5362 53.6224 16.4755 8.0257 20.5943 0.5 197 5 1 .9698 30. 1576 9.37 10 27.0435 0.54 14 54. 1399 21.9474 5.12 11 18.0 185 0.6 180 6 I .8020 16.4435 8.8652 28.8538 0.5467 54.6673 15.2143 5.67 16 23.4735 0.5594 55.938 I 13.1725 7.044 1 26.3450 0.537 1 53.7104 18.5455 8.1862 23.83 18 0.5288 52.8824
1 1.3313 2.0C98 9.7564 0.0004 3.5519
20.3904 5.73 19 20.025 1 0.5606 56.0603
3.3662 1.4 177 3. 1235 0.0 188 1.8846
C. 577 3 0.2431 0.5357 0.0032 0.3232
30. 1576 9.37 10 28.8538 0.6 180 61.8020
12.8565 4.5795 16.4479 0.5 197 51.9698
1 47
TABLE 12. Ifelo measurements, Group III: eleven worms from five h o s t s unU« (in mm.) same location as the Group I NUMBER LENGTH WIDTH ESOPHAGUS NERVE RING EX 20 1 2 . 4 9 5 0 0. 1 720 0 . 4 3 8 0 0 . 2 0 0 0 202 4.6 180 0 . 2 5 9 0 0 . 6 6 3 0 0.2 65 0 203 3.0 92 0 0. I860 0 . 5 5 0 0 0 . 2 4 8 0 204 4 . 8 0 0 0 0 . 4 5 2 0 0 . 8 C 2 0 0 .2 9 8 0 205 5 . 2 9 6 C 0.4 3 10 1 . 0 G 10 0 .3 4 5 0 206 4 . 7 2 0 C 0 .3 98 0 0 . 9 5 10 0.37 10 207 2 . 8 2 6 0 0 .2 05 0 0 . 4 5 4 0 0.24 10
VARIANCE 1 .2807 0.0 150 0 . 0 5 2 8 0.00 37
MEAN 3 .978 1 0.3 00 4 0 . 6 9 4 1 0.28 1 1
STANDARD DEVIATION 1.13 17 0. 1224 0 . 2 2 9 8 0 . 0 6 0 5
STANDARD ERRCR 0.42 7 7 0.04 6 3 0 . 0 8 6 8 0 .0 22 9 RANGE HIGH 5 . 2 9 6 0 0.4 52 0 1.00 10 0.37 10
LCW 2 . 4 9 5 0 0.1 72 0 0 . 4 3 8 0 0 .2 00 0
TABLE 13. Male derived statistios, Group III; eleven worms from five j (in mm.) same location as the Gr< ESOPHAGUS EXCRET.PORE NERVE RING TAIL BOD WORM BODY LENGTH BODY LENGTH BODY LENGTH BODY LENGTH NUMBER X 100 X 100 X 100 20 I 17. 555 I I I .9*459 8.0 I 60 5 .4 50 9 202 14.3569 10.0476 5.7 58 4 3.7029 203 17.7878 12.225 1 8 . 0 2 0 7 5. I 746 204 16.7083 10.0853 6 . 2 0 8 3 4 . 0 0 0 0 205 18.90 I 1 10.7628 6 .5 144 3.9464 206 20 . 148 3 I 1.0 169 7.8 60 2 4.2 16 1 207 ’ 6 • C 6 5 I 12. I 727 8 .5 28 0 4 .5 2 9 4
VAR I ANCE 3.58 18 0 .89 1 1 1 . 1825 0.4 33 3
MEAN 17.36014 I 1 . I 789 7 .2 69 4 4.43 15
STANDARD CEVIATICN 1.692 6 0 .9 44 C I .0874 0 . 6 5 8 3
STANDARD ERRCR 0 . 7 1 5 3 0 .3 56 8 0.4 110 0.24 88 RANGE HIGH 20 . 148 3 12.225 1 8 .5 2 8 0 5 .4 50 9 \:
LCW I *4 . 3 5 6 9 10.04 76 5.7 88 4 3 .7 02 9 l( o s t species captured in th e up I host. ING EXCRET.PORE SPICULES GUBERNACBLTJM TAIL )0 0.2980 0.2670 0. 1 150 0. 1360 10 0.4640 0.3 120 0. 1310 0.17 10 JO 0.3 780 0.3040 0.14 10 0. 1600 SO 0.4840 0.4080 0. 1520 0.1920 .0 0.5700 0.4640 0. 1660 0.2090 0 0.5200 0.4770 0. 169C 0. 1990 0 0.3440 0.2850 0. 1 180 0. 1280
7 0.0098 0.0078 0.0005 0.00 10
1 0.4369 0.3596 0.14 17 0. 1707
5 C.0991 0.088 1 0.02 17 0.0313
9 0.0375 0.0333 0.0082 0.0 118
0 0.5700 0.4770 0. 1690 0.2090
0 0.2980 0.2670 0. 1 150 0. 1280 r iv e h o st species captured in the ie Group I host. BODY LENGTH BODY LENGTH BODY LENGTH SPICULE TH WIDTH ESOPHAGUS TAIL GUBERNACULUM
9 14.5058 5.6963 18.3456 2.32 17 9 17.8301 6.9653 27.0058 2.38 17 6 16.6237 5.62 18 19.3250 2.1560 0 10.6 195 5.9850 25.0000 2.6842 4 12.2877 5.290 7 25.3397 2.7952 1 1 1 .8593 4.9632 23.7 186 2.8225 . + 13.7854 6.2247 22.078 1 2.4 153
i 6.7940 0.4286 10.34 19 0.0659
D 13.9302 5.82 10 22.9733 2.5 109
5 2.6065 0.6547 3.2159 0.2 567
0.9852 0.2474 1.2 155 0.0970
) 17.830 1 6.9653 27.0058 2.8225
> 10.6195 4.9632 18.3456 2.1560 48
TABLE 14. Female measurements, Group 111 : eleven worms from f i v e h o s t WCKM ( I n m m . ) same location as the Group NUMBER LENGTH WIDTH ESOPHAGUS NERVE RINGT E 30 I 6.IWC 0.3720 0.6900 0.2650 30? 5.75IC 0.3780 1.014C 0.3380 30 3 5.269C 0.4000 0.7480 0.2760 304 5. ICCC 0.2750 0.6230 0.2480
VARIANCE 0 . 2 1 4 3 0.0 C3 1 0 . 0 2 9 3 0.00 15
MEAN 5 .5 59 2 0 . 3 5 6 2 0 .7 68 7 0.28 17
STANDARD DEVIATION 0 . 4 6 3 0 0 . 0 5 5 5 0 . 1 7 1 3 0 .0 39 2
STANDARD ERRCR 0 . 2 3 1 5 0 .0 27 7 0 . 0 E 5 6 0 . 0 1 9 6 RANGE HIGH 6 . 1 1 7 C C .4 0C 0 1.0 140 0 .3 38 0 1
LOW 5. 1000 0.2 75 0 0 . 6 2 3 0 0 . 2 4 8 0 (
TABLE 1 5 . Female derived statistics, Group III: eleven worms f r o m f i v e h ( i n m m . ) s a m e l o c a t i o n a s t h e G r c E B O ^ A O 1"? EYCN'NT.PORE NERVE RING TAIL BOD- WORM BODY LENGTH BODY LENGTH BODY LENGTH BODY LENGTH 1! NUMBER X 1 0 0 X 100 X 100 X 100 30 1 I 1 .2 8 C C 8 .1 24 9 4.3522 3.4658 16 30 2 1 7.63 1 7 9 .1 11 5 5.8772 4.2601 1 5 303 14 . 1962 8 .7 8 7 2 5.2382 3.7958 13 304 12.2157 7 .6078 4.862 7 4.196 1 IE
VARIANCE 7 .8985 0. 4 5 3 C 0 . 4 2 2 3 0, 1 378 5
MEAN 13 . 8 30 V 8 . 4 C 7 9 5.0 7 76 3.9 29 4 15
STANDARD DEVIATION 2.8 104 0.6 73 1 0.6 49 9 - 0 . 3 7 1 2 2
STANDARDCRRCK 1 . 4 C 6 2 0.3 36 5 0 .3 24 9 0.18 56 1 RANGE HIGH 17.6317 9.1115 5 .8 77 2 4.260 1 18
L CW 1 J . 2 8 C C 7.60 7 8 4 . 3 3 2 2 3 .4 6 5 8 13 host species c a p tu r e d VULVA EGG EGG ?oup I host. ln %VA IK OP EXCRET.PORE TAIL TO ANTERIOR TO POSTERIOR LENGTH "'IDTH ) 0.4970 0.2 120 3.3440 2.7 7 30 0.0500 0.0300 ) 0.5240 0.2450 .3.2 170 2.5340 0.0590 0.04 3o ) 0.4630 0.2000 2.8300 2.4390 0.0660 0.0440 ) 0.3880 0.2 140 2.6970 2.4C30 0.0760 0.0460
* * C.0035 0.0004 0.0947 0.0278 0.000 1 0 .0 u • J 1 0 0.4680 0.2 177 3.0220 2.5372 0.C627 0.0407
C.C589 0.0 192 0.3078 0. 1666 0.0110 0.00 7 3
C.0294 0.0096 0. 1539 0.0833 0.C055 0.00 36
0.524C 0.2450 3.3440 2 . 773C 0.0760 0.0460
0.388C 0.2000 2.6970 2.4C30 0 .C50C 0.0300 Ive host species captured in the 3 Group I host. VULVA VTXVA BODY LENGTH BODY LENGTH BODY LENGTH TO ANTERIOR TO ANTERIOR H Y'lDTH ESOPHAGUS TAIL BODY LENGTH BODY LENGTH X 100 16.4435 8.8652 28.8530 0.5467 54.6673 15.?143 5.67 16 23.4735 0.5594 55.9381 13.1725 7.044 1 26.3450 0.537 1 53.7104 18.5455 8.1862 23.83 18 0.5288 52.8824
5.0636 1.9573 6.2627 0.0CG2 1. 7252
15.8439 7.44 18 25.6260 0.5430 54.2995
2.2502 1.3990 2.5025 0.0 13 1 1.3135
1.125 1 0.6995 1 .25 13 0.0066 0.6567
18.5455 8.8652 28.8538 0.5 594 55.9381
13.1725 5.67 16 2 3.4735 0.5288 52.8824 49 statistics or percentages, e.g., tail length/body length.
The ranges of measurements and derived statistics for the specimens in Group II, representing thirteen host species, are greater in all cases than those for specimens in Group I . However, the means of Group I are larger than those of Group II, with the exceptions of the length, width, and vulvar distances of the females and the width of the males. It had been noted that specimens of C. dukae from
Bufo americanus appeared to be larger than those from other host species.
There are no great differences between the variances of Group I and II, both composed of specimens from amphibia.
In males of these two groups, the gubernaculum and tail lengths showed the least variation, while body- length varied the most. In females of Groups I and II, length of the tail and width and length of the egg varied the least, the largest variation being that of body length. In general, as indicated by a comparison of variances, there is less variation among Group I specimens of C. dukae from a single host than within the other groups representing specimens from various hosts. This may be explained partly by the fact that parasites within a single host are more likely to have originated from the same parent worms, and therefore are more alike genetically, than parasites from different species of amphibian hosts taken from various localities.
The specimens in a single host were probably exposed to less 50 environmental variation than would have been found in a series of different hosts. Variances for male measurements in Group III are greater than those for Group I. The worms from these two groups were collected from hosts within an area which was approximately 400 meters wide by 800 meters long. It might be expected that these samples of C. dukae would be quite sim ilar. Since the worms of Group III came from several hosts, while those of Group I were obtained from one American Toad, the differences between these two groups may be due, at least in part, to host-induced variations.
The derived statistics vary more than the measure ments . This is due in part to the greater variation in worm length which is a factor in many of the ratios. The lowest ratio variances are, in the female, position of the vulva to body length and, in the male, length of the spicule to the length of the gubernaculum.
Included in the tables of derived statistics are ratios in addition to those used in comparisons with data from Anderson (i 960 ). These are body length/greatest width, body length/ esophagus length, and body length/tail length. The variance in the ratio of the body length is relatively low in both males and females.
Measurements of specimens of C. dukae from localities outside the state of Ohio were in close agreement with those of Ohio specimens. Measurements of C. dukae from Rana catesbeiana taken by W. A. Reid in M orrisville, North Carolina, fall within the ranges found in Ohio amphibians.
Of the specimens sent by the U.S. National Museum, those from Bufo quereieus from P t. Myers, Florida, and those from Rana sylvatica from White Lake, North Carolina, were within the size ranges of Ohio specimens. However, speci mens of C. dukae from the Yosemite toad, Bufo canorus, were slightly larger than Ohio specimens with respect to length of body, esophagus, tail, and in the position of the excretory pore and the nerve ring. The various size ratios for these worms were compared with those of Ohio specimens in order to check the significance of their size differences. In all cases the ratios for C. dukae from
B. canorus fall within the ranges for the same ratios for specimens from Ohio.
Although the specimens were not in good condition, since they had died within the digestive tract of the sala mander hosts rather than having been killed with hot alcohol,
C. dukae in Plethodon longicrus from North Carolina fell well within the range of measurement of Ohio specimens.
Ingles (1936), in noting the occurrence of C. dukae
^•n Triturus torosus, Rana aurora, and Bufo boreas in
California, mentioned the presence of a pair of ventrally located rosettes midway between the cloacal opening and the tip of the tail. However, after examining other specimens from the same host, he found that the presence of tubercles in this position was a variable character. Prom his figure 52 and discussion, it seems probable that these tail rosettes were comparable in size to those in the same specimens located anterior to the cloacal opening. While studying specimens of C. dukae furnished by the U.S. National Museum, the w riter observed tubercles around a pair of what other wise appeared to be simple papillae which were located midway between the cloacal opening and the tail tip. These were not large tubercles. Their size more closely approached that of the microtubercles of the complex papillae which were located anteriorly on this same specimen.
The specimens of C. dukae from Texas which were sent to the w riter by Dr. Paul D. Harwood agreed with C. dukae from Ohio both in the configuration of such important struc tures as spicules and gubernaculum and in the size of given structures. Although in his original description of these worms Harwood did not mention the small body papillae scattered over the entire cuticle of male and female speci mens, these papillae were present. The single median papillae located on the anterior cloacal lip of the male was double on a specimen from Bufo valliceps.
Observations on Cosmocercoides pulcher Wilkie, 1930
The examination of the holotype of Cosmocercoides dukae (Holl, 1928) revealed morphological details which were not discussed by Wilkie (1930) when he erected the genus
Cosmocercoides. Therefore, the writer felt that Wilkie’s 53 specimens should be re-examined in order to determine the relationship of C. dukae to C. pulcher which Wilkie had designated as the type of the genus.
Glycerine mounts of two intact females, two intact males, and the posterior half of a male positioned for a ventral view were received from the B ritish Museum for examination. One of the intact males had been damaged anterior to the esophageal bulb, and therefore measurements could not be made of this specimen in that body region.
With permission, a male and a female were removed from one slide for purposes of manipulation.
The body walls of these specimens were irregularly folded in varying degrees, either as a result of poor killing technique, originally, or caused by partial dehy dration at a later time. In addition, with the passage of time the specimens had turned a deep yellow-amber color.
Both of these factors obscured details, and increased the difficulty of measuring and studying the specimens. Treat ment with lactophenol did not clear them.
Wilkie did not state whether his figures of this species were averages, but he reported having ten males and sixty females available for study.
The number of complex papillae found by the w riter in the specimens of C. pulcher was 15 to 17 pairs in the three males observed. Wilkie reported 17 to 18 pairs. Since it is not known whether Wilkie examined all of the ten males 54 available to him, it can only be stated that the range in the number of complex papillae for W ilkie's specimens is at least 15 to 18. Hsu (1933) saw ten pairs of complex papillae in C. pulcher from the toad Bufo bufo Japonica in
C h in a.
Kung and Wu (1945) stated that their specimens of C. pulcher, taken from the same host in the same province in
China, showed no essential differences from those described by Hsu. Thus the number of pairs of complex papillae for
Kung and Wu's nematodes was probably about ten pairs. The range for the number of complex papillae of C. pulcher is ten to eighteen pairs, based on these reports. These measurements fall within the range of numbers of complex papillae for C. dukae,.
Additions to the Description of Cosmocercoides pulcher Wilkie, 1930
While examining type specimens of Cosmocercoides pulcher, observations were made on portions of the anatomy which supplement W ilkie's relatively short description.
Lateral alae were seen by the writer on each of
W ilkie's specimens which were loaned by the B ritish Museum
(Pig. 18).. These alae were difficult to detect since the bodies of the worms had softened and flattened considerably.
Alae were no| reported by Kung and Wu or by Hsu. These alae could be traced with difficulty for only a short dis tance because of the irregular folding of the body wall. On 55 the smaller of the two males, which measured 5.400 mm long, the posterior end of the ala was located 0.144 mm anterior to the cloacal opening. The anterior end of this ala was located 0.376 mm posterior to the oral opening. The alae were best defined on the larger male, which was 7.935 mm l o n g .
Wilkie did not mention either the nerve ring or the phasmids, although both were present in his specimens of
C. pulcher when they were examined by the present w riter.
The phasmids in the males were 0.075-0.086 mm from the tip of the tail, while in the females this distance was 0.069-
0.093 mm (Fig. 20). The position of the nerve; ring in this species is 0.418-0.440 mm for males and 0.450-0.550 for f e m a le s .
Cephalic papillae were neither drawn nor discussed in the description of C. pulcher by Wilkie. In a lateral view the U-shaped papillae of the outer circle can be seen; how ever, neither the small papillae of the inner circle nor the amphids were seen. Kung and Wu did not mention cephalic papillae, but Hsu illustrated a frontal view showing four double papillae in the outer circle, along with an amphid located laterally on each ventro-lateral lip (Fig. 32).
The four double papillae are comparable to the four horse shoe shaped papillae which Anderson (I960) illustrated for
C. dukae (Fig. 31). The amphids seen by Hsu may actually have been a pair of ventro-lateral papillae comparable to 56 those shown by Anderson for C. dukae. Frontal views of
C. pulcher were not prepared by the writer in the present study, since the type specimens would have been damaged had this been done.
Detailed drawings of the rosette-like complex papillae of C. pulcher have not been published. Although in the material furnished by the British Museum these structures were slightly distorted in some areas, in others they did not appear to be distorted. The latter were examined and drawings were made (Figs. 9, 10). It had been found that the rosettes of C. dukae from Ohio amphibians contained a circle of extremely minute tubercles within the circle of the larger tubercles (Fig. 8). However, these minute tuber cles were not found by the w riter in any of the complex papillae of C. pulcher. W. G. Inglis (personal communica tion) found no minute tubercles on the rosettes of C. tridens, nor did Khera (1958) illustrate them for C. m ultipapillata. However, an inner circle was found by
Ivanitsky (1940) for C. skrjabini (Table 18). The possibil ity exists that a softening of the cuticle of C. pulcher might have removed these minute projections.
On the type specimen of C. pulcher which was posi tioned for a ventral view, a single median papilla was found on the anterior cloacal lip (Fig. 24). This simple papilla did not appear to be doubled, although its base closely resembled that of the cloacal papilla on the cloacal lip of 57 male C. dukae of Ohio amphibians. This structure is not mentioned by Wilkie in his original description of C. pulcher, nor was it described in subsequent reports concern ing C. pulcher. The other two male type specimens studied were flattened on their sides and positioned so that such a papilla, if present, was obscured.
It is apparent to the writer from these observations that C. pulcher and C. dukae are more nearly alike than a comparison of the original descriptions could reveal. The principal discrepancy between these species is the differ ence in cephalic papillae. Considerable overlap was noted in the range of measurements of the spicules, gubernacula and e g g s .
An Emended Description of the Genus Cosmocercoides W ilkie, 1930
When the type specimens of Cosmocercoides pulcher and
C. dukae were examined in the present study, important details, which had not been reported previously, were revealed. Therefore, an emended description of the genus has been constructed incorporating these data with those given by Wilkie (1930) and Holl (1928).
Generic diagnosis
Cosmocercinae: narrow lateral alae present; very small papillae distributed over entire surface. Mouth with three small lips. Esophagus with short pharynx, slight isthmus, and a posterior bulb. Excretory system H-shaped, opening anterior to esophageal bulb. Male: Tail acutely tapering.
Two ventro-lateral, longitudinal rows of false plectanes, most of them pre-anal, on posterior one-fourth of body.
False plectanes purely cuticular with no internal skeleton and consisting of a large central papilla surrounded by two concentric circles of small tubercles. Simple papillae
on tail. Lateral alae ending at anal level. Slender spicules equal, arcuate, slightly flattened proximally at
capitulum; gubernaculum triangular. Female: tail long and pointed; vulva posterior to middle of body. Ovoviviparous within host; eggs, symmetrical; bluntly elliptical; thin
shelled. Parasites of large intestine of amphibians and
reptiles; mantle cavity and genital tract of terrestrial m o llu s c s .
Type: Cosmocercoides pulcher Wilkie, 1930, in "bull
frog" (Rana japonica?) Tokyo, Japan. Other
hosts: Bufo vulgaris, Japan and China; Rana
chensinensis, Eastern Siberia.
Observations on the Life History of Cosmocercoides dukae
A search of the literature revealed that Cosmocer
coides dukae has been recorded from thirty-five species of
amphibians, nine species of reptiles and seven species of
terrestrial gastropod molluscs. Anderson (i 960 ) showed that
snails and slugs can become infected with C. dukae by two different methods. First, infective larvae produced by
adult C. dukae living within the mantle cavity of a snail or
slug can migrate from that host and enter the mantle cavity
of another terrestrial gastropod. Second, Infective larvae which have been born within the mantle cavity of a snail or
slug may pass to the genital tract of the host by an unde
termined pathway and be incorporated within the shell of
the egg along with the egg cell. In this second method of
infection, a young gastropod mollusc is already parasitized
at the time it hatches from the egg.
One naturally wonders in what ways amphibia become
infected with this nematode. For example, is it possible
for amphibia to acquire the infection from terrestrial gas
tropod molluscs or do the latter ever acquire the infection
from amphibia? It would seem possible that either gastro
pods or amphibia might become infected by ingesting the
larval-contaminated feces of the other.
In order to determine whether gastropods could be
come infected with C. dukae which had been obtained from
amphibians, several dozen specimens of Rana pipiens were
collected from Ausable Point Marsh in Clinton County, New
York, in June, 1967 . Individual frogs were kept at ambient
laboratory temperatures in a small amount of water. The
water was changed daily and examined for larval and adult
stages of C. dukae lost from the frong by defecation. 60
Nematodes which had been collected were stored at 5 C until needed.
Each of 24 laboratory-reared land snails of the species Subulina octona was exposed to two larval stages of
C. dukae which measured between 600 and 1000 microns in length. This was accomplished by pipetting the larvae onto onto the withdrawn foot of the snail. A control series of
24 snails was treated identically to the experimental group, but was not exposed to larvae. Beginning seven days after the attempt to infect the snails, one control and one experimental snail were autopsied every other day and the results were recorded (Table 16). All control snails were nematode-free, while only seven (29 per cent) of the experi mental snails were nematode-free. However, nematodes of the latter group may have been lost in the watchglass in which the snails were housed. Only four specimens of C. dukae were located within the snail, but outside the mantle cavity.
Three of these four specimens were found in the rectum, while the fourth appeared to be lodged in the digestive gland. It is worth noting that seven of the seventeen snails contained two specimens each of C. dukae when they were examined.
Since Anderson (I960) had found C. dukae associated with the reproductive tract in land snails of Ontario, it seemed surprising that none were found there in Subulina
octona. This result might be partially explained by the 61
TABLE 16
INFECTION OF SNAILS WITH COSMOCERCOIDES DUKAE FROM AMPHIBIANS
Length of Nematodes (in microns) Site of Infection3, S n a il Day o f P re - P o s t M antle Number Autopsy i n f e c t io n in f e c t io n cavity Other
1 7 670,730 687 X 2 . 9 973,695 992,740 XX 3 11 862,830 884 X 4 13 795,658 negative 5 15 681,926 760 X
6 17 697,785 1120 X 1 19 848,708 814,920 X X 8 21 882,627 975 X 9 23 790,937 n e g a tiv e 10 25 898,891 1526,1480 XX
11 27 742,964 n e g a tiv e 12 29 824,893 1280,1393 XX 13 31 688,761 1026 X 14 33 752,857 n e g a tiv e 15 35 774,866 negative
16 37 864,722 1872,1320 X X 17 39 927,651 n e g a tiv e ..18 41 889,832 2016,2263 X X 19 43 660,710 1948,1570 XX 20 45 930,867 1525 X
21 47 825,645 1746 X 22 49 753,952 1931 X 23 51 943,702 1593 X 24 53 635,928 n e g a tiv e
aSymbol * = one nematode. 62 fact that different host species are involved. In addi tion, while culturing Subulina octona in the laboratory, the writer observed that these snails were less active when they were housed in small chambers.
Those specimens of C. dukae which were recovered later in the experiment shoVed a considerable increase in size. The six specimens which were larger than 1700 microns had reached sexual m aturity.
Cosmocercoides dukae from amphibians molts and matures in Subulina octona, a land snail belonging to the family Achatinidae. In order to approximate more closely the conditions of contact between the parasite and host in nature, each of a series of twelve snails which had been starved for two days was placed in a covered moist watch- glass. Feces from a frog heavily infected with C. dukae was placed on a 10 mm. square of lettuce in each watchglass.
Beginning with the day after this feeding one snail was autopsied each day from this series as well as the control series and the results were recorded (Table 17)* These showed a high percentage of parasitize4 snails with only one individual free of nematodes. After just one contact with infected frog feces, snails of the genus Subulina retain specimens of C. dukae for at least twelve days under labora tory conditions. The genus Opeas, also a member of the family
Achatinidae, was found naturally infected with Cosmocer coides by Chitwood and Chitwood (1937). 63 TABLE 17
INFECTION OF SNAILS WITH COSMOCERCOIDES DUKAE BY INGESTION OF FROG FECES
Day o f A utopsy, Worm Recovery Site S n a il a f t e r I n t e s t i n a l M antle Number fe e d in g t r a c t c a v ity O ther 1 1 3 la rv a e 4 larvae negative
2 2 n e g a tiv e n e g a tiv e n e g a tiv e
3 3 n e g a tiv e 1 la r v a n e g a tiv e
4 4 7 larvae 6 larvae n e g a tiv e
5 5 2 la rv a e 1 larva negative
6 6 1 la r v a negative negative
7 7 2 la rv a e n e g a tiv e n e g a tiv e
8 8 9 la rv a e 7 la rv a e 1 larva, in kidney?
9 9 n e g a tiv e 4 larvae negative 10 10 negative 12 larvae & n e g a tiv e 1 adult female
11 11 3 la rv a e n e g a tiv e n e g a tiv e
12 12 negative 2 larvae & n e g a tiv e 1 adult female 64
Therefore, from the present study and from the work of
Ogren (1953, 1959) and Anderson (I960), terrestrial molluscs may become infected in at least three ways, viz., by being infected while s till developing as embryos within the egg shell; by contact with the foot or slime trail of an infected snail; and by contact with C. dukae contained in amphibian feces which is ingested by them.
Observations made during this study also indicate that amphibian hosts may affect the life cycle in still another way. Over-wintering may be enhanced in cooler geographical regions where host species are not active during the entire eyear by the fact that C. dukae can exist for long periods of time within a chilled host animal. Several hosts kept for an extended period of time at
5 C without food contained C. dukae when autopsized. One of these, an American Toad which was captured in Laurel Town ship, Hocking County on November 23, 1963, died on April 17,
1964, 147 days (21 weeks) later. Two specimens of C. dukae, one male and one female, were found in the large intestine.
During the time this toad was confined, the container in which it was held was checked at two to five day intervals for the presence of specimens that might have been passed in its feces. Dead adults and both active and dead larvae were found. A second American Toad, captured in Laurel Township on the same date, died 159 days later on April 29, 1964. The large intestine of this host harbored six adult females and nine adult males. In addition to these two toads, one individual bullfrog, Rana catesbeiana, from Westfield
Township, Morrow County, was kept at 5 C in the laboratory for sixteen weeks after it was captured. It contained one female C. dukae when autopsied. Three of four bullfrongs from the same location kept under the same conditions for eight weeks harbored from one to seven cosmocercids. These observations, especially those concerning the toads, sug gest that C. dukae could possibly over-winter within amphibian hosts .
Anderson (i 960 ) observed th&t about one-fourth of
32 amphibians were s till infected when they were autopsied a month or more after being held in the laboratory ". . .a t such low temperatures that defaecation was infrequent."
In addition, both larval and adult worms lived for long periods of time in saline and in tapwater. Adult male and female specimens in Ringer's "cold" were placed in a refrigerator at 5 C on October 30, 1963 . One fem ale i n d i vidual was s till alive on February 9, 1964, fourteen weeks and four days later. When recently laid eggs from a female worm were placed in Syracuse watchglasses that were half filled with saline, and kept at 5 C in a refrigerator, living hatched larvae were still active five weeks later.
When specimens of £. dukae were removed from hosts, these specimens were examined in all cases to see whether the females were oviparous or ovoviviparous. Hatched larvae were seen In utero on a number of occasions, and several were seen ejected from the reproductive tract. The worms were removed very rapidly from the large intestine of a small American Toad and subsequently passed through several saline washings to eliminate larvae which might already have been in the intestine of the host. The repro ductive tract of one female was removed quickly and the uteri were found to contain five first-stage larvae that had hatched within the tract. Thus C. dukae appears to be ovoviviparous, at least for a short time outside the body of the host. This may also be the rule within the large intestine of amphibian hosts. However, specimens of C. dukae placed in either water or saline usually lay many eggs, of which one-half to three-fourths usually hatch.
These observations on the time of hatching agree with those of Ogren (1953, 1959) and Anderson (i 960 ).
During this study the food content of the alimentary tract of the amphibian hosts was noted, although no detailed investigation was made to determine the diet of these animals. Portions of chitinized arthropod skeletons were commonly found. Several larger toads contained partially digested land snails in the stomach. By comparing the shell fragments with a series of shells of Ohio snails collected by the writer, these partially digested snails, were identified as Mesomphix Rafinesque, 1831. Other snails found in areas where toads were captured were identified as (1) Mesodon thyroidus (Say) (Synonyms: Helix
thyroidus Say; Helix throides Say; Mesodon thyroides Say;
and Polygyra thyroides (Say) and (2) Triodopsis trldentata
(Say) (Synonyms: Helix 'trldentata Say and Polygyra tridentata
Say). The nomenclature follows that of Taft (1961). DISCUSSION
In evaluating the taxonomic position of Cosmocer coides dukae, it seems advisable first to mention briefly the morphological characters considered by investigators of this genus. The possession of an esophagus with a posterior bulb, a simple intestine without a diverticulum, and, in the female, a finely pointed tail allow the various species in the genus Cosmocercoides to be called oxyuroids in the broad sense as included in the superfamily Oxyuroidea by Yorke and Maplestone (1926). The female possesses two ovaries, an anterior and a posterior one, and the vulva is located posterior to the middle of the body. In addition, the male possesses two spicules and a gubernaculum, features which were used by R ailliet (1916) to erect the subfamily Cosmocercinae. These characters, along with two ventro-lateral rows of pseudo-plectanes (rosettes) in the male, identify the genus Cosmocercoides and separate it from closely related genera. The identification of a female belonging to the genus is difficult if obtained from a host lacking male parasites„
During the course of this study, the literature revealed that seven species of Cosmocercoides had been described. These were C. pulcher Wilkie, 1930; C. tridens
68 69 Wilkie, 1930; C. dukae (Holl, 1928); C. bufonis Karve,
1944, C. multipapillata Khera, 1948; C. skrjabini (Ivan itsky, 1940); and C. timofe.1ovoi (Skarbilovich, 1950).
When the characteristics of these species are tabu lated, it will be noted that many of the criteria used in separating them are based on only minute size differences
(Table 18). While it is conceivable that such differences might serve to- establish separation into the various species that are listed, it is disturbing to note that when the ranges of size are considered there is considerable overlapping, so that the range in measurements of one species often falls within the range of measurements of another.
It is to be expected that the grater the number of specimens of a species that are measured, the greater may be the range in sizes represented. It seems somewhat mis leading to compare the size range of a given structure where many measurements have been made with the size range of a similar structure, in what has been desdribed as a different species, where only a few measurements have been made. This is especially puzzling when the range repre sented by a few measurements fall either within or over laps by only several hundredths of a millimeter the range represented by many measurements of what is thought to be a different species. T*3I£ 1R
A OCia’AJtlSON OF TICE SPECIES OF TiiiC GH.liS CCCLUCMCulDLS (ICe&sureroents in m illixiciero)
C, dukae C. pulohor C. fcridens C. m ultipapillata C. bufonis C. skrjabini C. *wir-ofejcvoi
* MALE
L e n g t h 1 .6 5 - 5 . 4 G 7 5 , 4 - 7 . 6 4 . 3 5 . 8 - 6 .4 4 . 5 - 4 . 9 2 .... j . V l D i a m e te r 0,12-0.60 0.260-0.60 0 . 4 4 0.36-0.39 0.35-0.425 .... 0.Lk$-0.61i
FEMALE
Length 1.66-6.734 7.2-11,0 .... 6 . S 5 - 7 . 1 4 5 , 1 5 - 6 .3 0 .... 5 . 1 3 - 7 . t:5 D i a m e te r 0.133-0.60 0.40-0.610 0.40-0.417 0.45-0.56 0■5*,—o .7 0
ESOPHAGUS
Length in Male . 0,350-1.023 1 . 0 9 - 1 . 3 0 0 . 9 3 0.930-0.960 0 . 6 8 - 0 . 6 6 .... u , 6 1 - 0 . 7 3 Length in Female 0.330-1.306 1 . 2 2 - 1 . 4 6 0 1.0-1.1 0.65-1.06 ...» u .A * ? - o .t6
E^ornAnEAL *utl8
Diameter in Mile 0.060-0.133 0.13-0.140 0.12 • 0.75-0.60 0.125-0.135 * • • .' Diameter in Fem ie 0.065-0.152 0.126-0.245 0.135-0.145 0.15-0.16 .... UU/.U
TAIL
Length ir Mile 0.104-0,265 0.208-0,318 0.20 .... 0 . 2 0 - 0 . 2 4 u . 2 V - u . 3 5 Length in Female 0.120-0.324 0.332-0.520 .... 0.22-0.25 0 . 2 3 - 0 . 2 6 ....
EXCRETORY PORE (From an te rio r end)
Position in ifcle 0.240-0,583 0.G83-0.782 0,52 0.57-0,51 0.36-0.37 • .... u . 3 7 - 0 . 3 6 Position in Female 0.140-0.564 . 0.74-1.0 .... 0,63-0.55 0 . 5 0 - 0 . 6 8 ....
VULVA
Position-from anterior end 0.R60-3.R05 3.954-5 ,00 . . • ...... Position from ta ll tip 0.700-3.641 3 . 3 4 - 5 . 0 4 2 . 9 - 3 ,3 2 . 2 0 - 2 . 7 0
QUBSRMACULUV
L e n g t h 0.070-0.163 0.144-0.180 0.14 G .1 3 - 0 , 1 4 0 . 1 2 - 0 . 1 4 5 0 . 2 1
SPICULE
L e n g th U .2G U -u .*»76 0.168-0,333 0.53 0 . 2 0 - 0 . 2 4 0.19-0.26 0.212 0 . 3 0 - 0 . 3 5
EGG
L e n g th 0.044-0.nno 0.045-0.082 .... O.OG72-0.0724 0.075-0.000 .... *111 D i a m e te r . 0 . 0 2 5 - 0 . 0 5 0 0.029-0.045 0,0430-0.0457 0.040-0.0*16 0 a 0 5 - . : . o 77
-< 3 i O T.\2I£ lc (ccntd)
G. 'hi]ja»> C # n u lc tH T C. trld*'n3 C. r.iiltjpapitlata C . b u f o n i s C. nhrj.-.M ni C. tia.oi'ejovoi
8IVPI.5 PAPILIAE
Dorsal U p of Itele 2 p l u s 2 2 . . . . 2 2 ...... V #ntro-l*ter*l lip* of ’.tel# 2 plus 2, eaoh 1 on e a o h 2 o n e a o h 2 on e a c h
D o r r a l l i p o'* F e m a le 2 pl*-8 2 2 2 2 .... V entro-lateral lie* of Fermi# 2 plus 2, eaoh 1 on e a o h • • • 2 on e a o h 2 o n e a o h
Cervioal panillae of 'tele Level of nerve ring .... Cervioal papilla# of Female ...... • • • • .... Level of nerve ring
Foat-anal papilla# of Male 4 p a i r * 6-13 pairs .... 5 p a i r s 10 pairs-plus .... fc-7 pairs Poit-anal papilla# of Female 1 -3 p a i r * 9 p a i r * N one 2 p a i r * £ - 7 p a i r s
Cloaoal papilla# of Mile 1-2, anterior lip Ono None 1, each lip Nono Cloaoal papilla# of Female Won# Mon# • • • • Kona None » c n o
Other body papilla# of 'tel# Entire body Entire body .... Entire body .... Entire* body Ot'*er bo CO.'TLFX PA PIT LAE (Rosettes or Pseudoplootanos) Pro-anal of Male 6-23 pair* 7-16 pairs 1 1 p a i r s 10-14 pairs 9-13 pairs .... 16-2J pairs Pr*-*nal of Female N one Non# Nono N ono Nono L o n e Ad-anal of ’tel# 2 p a i r s 2 p a i r s N one 1 p a i r 1 p a i r Ad-anal of Female N o re N one Nono N one None • « . • . . . . Peat-anal of Male 1 p a i r 3 p a i r s 3 p a i r s 3 p « « irs 3 p a i r s .... ro»t-*p»l of Fenele N one Non# N one N one None :::: Total ’■osettes n1’ Itel# 9-26 pair* 10-17 pairs 14 p a i r s iA-lfc pairs 13-1? pairs 7 - 9 p a i r s lo-ib* pairs Total rorettOF or Female N one None ?,o no None L o n e GSOGpAHUCAI nirTRlBrTI"H North -Amorica Japun, China, Siberia J a p a n I n d i a I n d i a . R u s s i a K i r g h i z HC5T5 G a s tr o p o d s A m p h ib ia n s A m p h ib ia n s A m p h ib ia n s A m p h ib ia n s Asp.hibian* a m p h i b i a n s A m p h ib ia n s R e p t i l e s Many taxonomists have commented upon the use of size as a criterion for distinguishing between taxa. M. B. Chitwood (1957) notes that size alone is useless except as a relative concept since it varies with age, physical condi tion, metabolic rate and food supply of the host. She states that the "... concept of species must be broadened by study of more specimens for variations and influence of geographical and ecological factors." The sizes for the type species, C. pulcher, represent measurements by Wilkie of sixty females, but only ten males. The additional measurements by Hsu of five females and two males and of Kung and Wu of two females and two males bring the total number of measurements for this species to sixty-seven for the females, but only fourteen for the males. In view of the great variability in ranges of measurement which have been shown for C. dukae, it is quite possible that the ranges for C. pulcher would be larger if more measurements were made. It is clear that little reliance can be placed on the diagnostic measurements of a species such as C. tridens which is represented by only a single specimen, in this case a male. In this species the spicule is given by Wilkie as 0.53 millimeters in length, which he indicates as a distinguishing character for it; however, this represents the measurement of but only one worm since the single specimen is the only one that has ever been seen of this species. On the other hand, the length of its spicule which is 0.53 millimeters is only 0.21 millimeters longer than the largest spicule measured for C. pulcher. The difference in size of the longest and shortest spicule in the size range of C. pulcher is 0.155 millimeters. Both C. pulcher and C. tridens were taken from hosts in the same geographical area, Tokyo, Japan, although they were obtained from different types of host animals. The writer was unable to secure a loan of the single specimen of C. tridens but he was furnished additional data concerning it by Inglis (personal communication). The species C. skrjabini from Russia is known from only male specimens, and it was not possible to obtain these for the purpose of checking with the original description. A request for the loan of specimens from India of C. multipapillata ajid_£. bufonis brought no response, and here again the writer had to resort to compiling the data made available from the literature covering these species. As another specific example of the lack of data, the cephalic papillae have been illustrated by eri face views for only two of the six species in the genus. Hsu (1933) illustrated the papillae for C. pulcher (Pig. 32). They have been shown for C. dukaeby Chitwood and Chitwood (1937b), by Ogren (1953), by Anderson ( i 960 ), and by the writer in the present study (Figs. 30, 31, 28). Kreis (1932) stated that Trionchonema rusticum, now a synonym of C. dukae, does not possess cephalic papillae and his illustration of the anterior end of a male includes no papillae (Fig. 29). Karve (1944) illustrated only a lateral view of the cephalic region of C. bufonis, in which he found two papillae per lip. The large size of the papillae he illustrates in dicates that they probably correspond to the large lip p a p illa e shown by Anderson and not to the two minute papillae seen by Khera (1958) on the lips of C. multi- papillata. Cosmocercoides bufonis may have these six minute papillae, since they are extremely small and could easily have been overlooked by Karve. Although it seems unlikely that Khera would have missed seeing the larger papillae, the writer noted that often it was not possible to detect these s tru c tu re s when viewed l a t e r a l ly . The cephalic p a p illa e of C. dukae from some Ohio amphibians agree in number and posi tion with those found by Anderson (I960) in C. dukae from molluscs in Ontario. It seems more likely that all species of the genus Cosmocercoides have cephalic papillae corres ponding to those which were found in C. dukae in the present study and a lso by Anderson. Khera (1958) stated that C. multipapillata differs from C. variabilis and C. bufonis in the presence of small simple papillae over the entire surface of C. multi papillata. Since Karve noted small simple papillae in the pre-anal, as well as the post-anal regions,and since the writer observed these micropapillae over the entire surface of specimens of C. variabilis (= C. dukae) loaned by Harwood, this difference does not exist. Karve and Khera felt that C. bufonis and C. multipapillata, respec tively, differed from C. variabilis in the configuration of the lips. This feature was found to vary considerably in specimens of C. dukae from Ohio amphibians and the writer feels that little weight can be given to it for species diagnosis within the genus Cosmocercoides. Variations in the contraction of esophageal and body wall musculature in the cephalic region at the time of fixation affect lip configuration. Although neither Karve nor Khera recorded the number of specimens measured in their studies, the rather narrow ranges would seem to indicate that this number was not large. The writer concludes that the only differences between C. bufonis and C. multipapillata are the simple post-anal papillae of the males and the presence of cloacal lip papillae in C. bufonis. Slides of two female and three male specimens of C. pulcher were loaned by the British Museum of Natural History for checking the original description of this species. An analysis revealed that they had lateral alae, a fact which had not been reported by Wilkie. Therefore, in this respect C. pulcher agrees with C. dukae, C. multi papillata, and C. bufonis. Since material for C. tridens and C. skrjabini was not available, a positive statement concerning these two species cannot be made, although it 76 seems highly probable that these also have lateral alae. Table 18, which is presented as a key source, repre sents a composite of data collected, whenever possible, by examining available specimens and/or obtaining these data by private correspondence and by referring to all available publications on the species of the genus Cosmocercoides. Prom these data it is the opinion of the writer that five of the forms, viz., C. pulcher, C. multipapillata, C. bufonis, C. timofejovoi and C. dukae should be accepted as valid species, with the possible addition of C. skrjabini, although the latter species cannot be evaluated fully until more material consisting of both males and females is avail able. Within the limitation of the data presently available on the various species assigned to the genus Cosmocercoides, the main character for distinguishing species is the number of simple papillae on the male tail. The writer further suggests that the specimen referred to as C. tridens might possibly by a synonym of C. pulcher since its characters, with the single exception of a slight difference in size of the male spicule, agree in general with those given for C* pulcher. Again, any justification of it as a valid species must depend on additional numbers of individuals with characters different than those given for any of the other four, or possibly five, species. Skrjabin et a l. (1961) have discussed the confusion which exists in the taxonomy of the cosmocercids. As an example, they note that over the span of a century not a single European investigator observed a mature specimen of Oxysomatium longispiculum, while during this same period of tim e, O'xysomatium brevicaudatum was observed a g reat many times. They conclude that the only inference to be drawn is that only one species exists in European hosts. In a similar way, it is quite possible to draw the same conclu sion with respect to Cosmocercoides pulcher and Cosmocercoides tridens. The latter has been observed only once—a single male specimen—over a period of thirty-eight years, while the former has been recorded a number of tim e s. Mayr, Linsley and Usinger (1953) state that taxonom ists have been too willing to split an established species and have thus cluttered the literature with synonyms. They note that it has been estimated that more than half of all synonymy is due to underestim ation of in d iv id u a l v a r i ation within populations. The writer is convinced that valid and usable descrip tions of cosmocercid nematodes must include detailed accounts and drawings of the adult male anatomy based upon a large number of specimens. This opinion, which is based upon this study, is supported by opinions received through personal conversations with the late C. C. Walton, and with H. P. Hsu and M. B. Chitwood. Usable descriptions must include complete cephalic lip detail, lateral and dorsal views of the gubernaculum, and a full account of the complex and simple papillae, as well as other measurements included in Tables 4 through 15 and Table 18. This information is available, with the completion of this study, for only one species in the genus Cosmocercoides, namely, C. dukae. If similar data are compiled in the future for Cosmocercoides pulcher in Europe and Asia, and if the degree of variability for C. pulcher is comparable to that of C. dukae, the writer feels that this genus will probably contain no more than two species. However, such a statement would be premature at the present time. SUMMARY Eighteen species of amphibians and reptiles were examined during this research. While none of the reptiles were infected with Cosmocercoides dukae, 19 per cent of the amphibians had this parasite. These hosts were obtained from Fairfield, Franklin, Hocking, Morrow, Sandusky and Vinton Counties. Localities outside Ohio which were represented in this study were Florida, North Carolina, Kentucky, Texas, and Ontario in Canada. Including speci mens loaned by other investigators and the U.S. National Museum, twenty host species were represented. One frog, Pseudacris brachyphona, and two salamanders, Eurycea bislineata and Plethodon long!crus, were reported as hosts for C. dukae for the first time. Variability in measurements and in ratios derived from measurements were described, and considerably overlap of ranges was noted. Certain details of the anatomy of C. dukae were discussed and the adult of this species from Ohio amphibians was described.^ These results, coupled with the fact that no striking differences in the shapes of struc tures were found, lead the writer to the conclusion that at the present time there has been only one species belonging 79 80 to the genus Cosmocercoides seen or described in North America, namely, C. dukae. The land s n a il Subulina octona was in fe c te d by placing third stage larvae of C. dukae on the withdrawn foot of one group and by allowing another group to feed on lettuce onto which feces of frogs infected with C. dukae had been placed. Thus, a snail cannot only be infected while still a young embryo within the egg shell, and by contact with the slime trail of an infected snail, but also by contact with amphibian feces. The results of the snail infection experiments indicate the possibility that "C. dukae may be passed more frequently from amphibian to mollusc hosts than heretofore thought possible. Cotypes of Cosmocercoides pulcher were examined and additions to the description of this species were made. The presence of lateral alae and a papilla on the anterior cloacal lip were reported for the first time. The various species of the genus Cosmocercoides were compared,and the basis for distinguishing them was dis cussed. Cosmocercoides pulcher, C. timofejovoi, C. multi papillata, C. bufonis, and C. dukae were accepted as valid species. More information is required before the status of C. skrjabini can be evaluated. It is suggested that C. tridens be considered as a synonym of C. pulcher since a slight difference in the spicule length is scant evidence for maintaining a species within a genus which exhibits marked size variations in so many characters within a single species. APPENDIX HOSTS REPORTED FOR COSMOCERC01DES DUKAE #Infected host species examined in this study Amphibia Frogs Acris gryllus Le Conte Reference: Walton (1938) Hyla cinerea Reference Walton (1946b) Hyla c r u c ife r Weid Reference: Brandt (1936) Rankin (19*15) *!• squirella. Latreille Reference: Harwood (1930) *Pseudacris brachyphona (Cope) Pseudacris brimleyi Brandt and Walker Reference: Brandt (1936) Walton (19*l6a) P. nigrita triseriata (Wied) Reference: Walton (19*l6a) P. t r i s e r i a t a Weid Reference: Harwood (1930) Rana a re o la ta (B aird and G ir., 1852 Reference: Harwood (1930) R. aurora (Baird and Girard) Reference: Ingles (1936) 82 *R. catesb eian a Shaw, 1802 Reference: Harwood (1932) Brandt (1936) Rankin (1945) Anderson (i 960 ) *R. clamltans Latreille, 1802 Reference: Harwood (1932) Rankin (1945) Anderson (i 960 ) R. palustris Le Conte Reference: Harwood (1930) *R. plpiens Schreber, 1782 Reference: Anderson (i 960 ) R. septentrlonalls Baird Reference: Anderson (i 960 ) R. sphenocephala (Cope, 1886) Stejneger and Barbour, 1917 Reference: Harwood (1930,1940) Brandt (1936) *R. s y lv a tic a Le Conte, 1825 Reference: Harwood (1930) Toads *Bufo americanus Holbrook, 1836 Reference: Anderson ( i 960 ) B. boreas (Baird and Girard) Reference: Ingles (1936 Frandsen and Grundmann ( i 960 ) B. canorus Camp Reference: Walton (1941) B. fowleri Hinckley , 1882 Reference: Brandt (1936) §.• quercicus Holbrook Reference: Walton (1938) terrestris (Bonnaterre) . - Reference: Harwood (1932) §.• valliceps Wiegmann, 1833 Reference: Harwood (1930) 84 *B. woodhousii fowleri (Hinckley) Gastrophryne areolata Reference: Harwood (1930) G. carolinensis (Strecker) Reference: Walton (1950a) *Scaphiopus holbrooki (Harlan) Reference: Brandt (1936) Salamanders Ambystoma Jeffersonianurn Green Reference: Anderson (I960) A* opacum (G ravenhorst) Reference: Rankin (1937) A. talpoideum (Holbrook) Reference: Harwood (1932) Rankin (1937) A. texanum M atthes, 1855 (= Ambystoma microstomum Cope, 1861 = Ambystoma microstomom (Cope, 1861) S tejn eg er and Barbour, 1917 Reference: Harwood (1930) Rankin (1937) *Desmognathus f . fuscus Rafinesque Reference: Rankin (1937) Pischthal (1955b) *Eurycea bislineata Plethodon cinereus (Green) Reference: Rankin (1937, 1945) glutinosus (Green) Reference: Rankin (1937) *P. longicrus Adler and Dennis, 1962 Reference: Adler and Dennis (1962) T ritu ru s m erid lo n alis (Cope, 1880), Dunn, 1918 Reference: Harwood (1930) T. torosus Reference: Ingles (1936) Lehman (1954) T. viridescens Raflinesque, 1820 Reference: Holl (1928) Mueller (1932) Rankin (1937, 19^5) Fischthal (1955 a and M ollusca Slugs Deroceras graeile Rafinesque Reference: Ogren (1959b) Anderson (I960) Deroceras sp. Reference: Ogren (1953) S nails Ashmunella rhyssa edentata Reference: Ogren (1953) Discus cronkheitel (Newcombe) Reference: Anderson (i 960 ) Polygyra fosteri Reference: Ogren (1953) Retine11a sp. Reference: Ogren (1953) Zonitoides arborea (Say) Reference: Anderson (i 960 ) R ep tiles Lizards Eumeces fasciatus R eference: Harwood (1932) Ophisaurus ventralia Reference: Harwood (1930) Leiolopisma laterale R eference: Harwood (1930) Snakes Heterodon contortrix Reference: Harwood (1930) Micrurus fulvlus Reference: Harwood (1930) Storerla dekayl Reference: Harwood (1930) S. occlpitomaculata Reference: Rankin (19^5) T u rtles Terrapene Carolina Reference: Harwood (1930) Rausch (19*17) T . o rn a ta Reference: Harwood (1930) LITERATURE CITED Adler, K. K. and D. M. Dennis. 1962 . Plethodon longicrus a new salamander (Amphibia: Plethodontidae) from North Carolina. Spec. Publ. Ohio Herpetol. Soc. 4: 1-14. A nderson, R. C. 1958. Methode pour l'examen des nematodes en vue apical. Ann. Parasitol. Humaine et Comparee 33:171-172. Anderson, R. C. i 9 60 . On the development and transmission of Cosmocercoides dukae of terrestrial molluscs in Ontario. Canadian J. Zool. 38(4):801-825. Ballesteros-Marquez, A. 1945. Revision de la Familia Cosmocercidae Travassos, 1925. Rev. Iberica Parasitol., Granada, Tomo Extraordinario 1945, pp. 150-180. Brandt, B. B. 1936. Parasites of certain North Carolina Salientia. Ecol. Monographs. 6:492-532. Chabaud, A. G. and E. R. Brygoo. 1958. Description et cycle evolutiv dfAplectana courdurieri n. sp. (Nematoda: Cosmocercidae). Mem. Inst. Sci. Madagascar. 12A: 159-176, 5 fig. Chitwood, G. B. 1933a. Trionchonema Kreis, 1933 synonym of Cosmocercoides Wilkie, 1930. J. Parasitol. 19:243. Chitwood, B. G. 1933b. On some nematodes of the super family Rhabditoidea and their status as parasites of reptiles and amphibians. J. Wash.Acad. Sci. 23(11): 508- 520 . Chitwood, B. G. and M. B. Chitwood. 1937a. Snails as hosts and carriers of nematodes and nematomorpha. The Nautilus. 50:130-135 (1937). Chitw ood, B. G. and M. B. Chitw ood. 1937b. Cosmocercoidae trainsferred from Oxyuroidea to Ascaridoidea. Intro duction to Nematology . 1937, p. 50. 87 88 Chitwood, M. B. 1957* I n tr a s p e c ific v a ria tio n in p a r a s itic nematodes. Systematic Zoology 6(1): 19-23. Pischthal, J. H. 1955a. Ecology of worm parasites in South-central New York salamanders. Amer. Midland Naturalist 53:176-183. Pischthal, J. H. 1955b. Helminths of salamanders from Promised Land State Forest Park, Pennsylvania. Proc. Helm. Soc. Wash. 22:46-48. Prandsen, J. C. and A. W. Grundmann, i 9 60 . The p a ra s ite s of some amphibians of Utah. J. Parasitol. 46:678. Harwood, P. D. 1930. New species of Oxysomatium with remarks on the genera Oxysomatium and Aplectana. J. Parasitol. 17:61-73. Harwood, P. D. 1932. The helminths parasitic in the Am phibia and Reptilia of Houston, Texas and vicinity. Proc. U.S. Nat. Mus. 81(17):1-71. Holl, Pred, J. 1928. Two new nematode parasites. J. Elisha Mitchell Scientific Soc. 43:184-186.1 pi. Hsu, H. P. 1933. On some parasitic nematodes collected in China. Parasitol. 24:512-541, illus. Ingles, L. G. 1936. Worm parasites of California Amphibia. Trans. Amer. M icr. Soc. 55:73-92, i l l u s . Ivanitzky, S. V. 1940. (Additions to helminth fauna of vertebrates of Ukraine) (fauna of cestodes, nematodes, and acanthocephalans) (Russian text). Sb. Tr. Kharkov Vet. Inst. 19:129-155. Karve, J. N. 1944. On a small collection of parasitic nematodes from Anura. Proc. Indian Acad. Sci. B19: 71-77. 5 figs. Khera, S. 1958. On a new species of Cosmocercoides♦ Indian J. Helminthology 10(1):6-12. 7 figs. Kreis, H. S. 1932. Trionchonema rusticum n.g., n. sp., a parasitic nematode from the land snail, Polygyra c a p ic o la . Bland. (H e lic id a e ). Trans. Amer. M icros. Soc. 51:48-56. Kung, and Wu. 1945. Cosmocercoides pulcher Wilkie in Bufo bufo asiaticus, China. SInesia, Nanking. 16:75=76. 89 Lehman, D. L. 195*1. Some helminths of west coast urodeles. J. Parasitol. 40:231. Mayr, E., E. G. Linsley and R. L. Usinger. 1953. Methods and principles of systematic zoology. McGraw-Hill Book Co., 328 p. Milogradova, G. P. and A. A. Spasskii. 1957. (Helminth fauna of Anura in eastern Liberia) (Russian text). Tezisy Dokl. Nauchn. Konf. Vsesoiuz. Obsh. Gelmint. Posviash. 40 g. Oktiabr. Sotsial Revoliuts., 11-15 Dec., Chast 1, p. 200. Mueller, J. P. 1932. Capillaria tenua, a new species of nematode parasitic in the newt, Triturus viridescens. Trans. Amer. M icro. Soc. 51 (4):266. Ogren, R. E. 1953. Cosmocercoides dukae (Holl) redescr. from Polygyra fosteri, Retine11a sp. Ashmunella rhyssa ed en tata and Deroceras s p ., N. America. Trans. Amer. Micr. Soc. 72:67-91. figs. Ogren, R. E. 1959a. The nematode Cosmocercoides dukae as a parasite of the slug. Proc. Pennsylvania Acad, of Sci. 33:236-241. Ogren, R. E. 1959b. The nematode Cosmocercoides dukae as a parasite of the slug. (Abstract) J. Parasitol. 45 (4, Sect. 2):45. Rankin, J. S., Jr. 1937. An ecological study of parasites of some North Carolina salamanders. Ecol. Monographs 7:170-269. Rankin, J. S., Jr. 1945. An ecological study of the helminth parasites of amphibians and reptiles of western Massachusetts and vicinity. J. Parasitol. 31:142-150. Rausch, R. 1947. Observations on some helminths parasitic in Ohio turtles. Amer. Midland Naturalist 38:434-442. Skrjabin, K. I., N. P. Schichobalow, and A. A. Mosgowoi. 1951. Oxyurata und Ascaridata, in: Opredelitil parazititchskich Nematoda, Bd. 2. Skrjabin, K. I., N. P. Shikhobalova, and E. A. Lagodovskaya. 1961. (Principles of nematology, edited by K. I. Skrjabin. Vol. X Oxyurata of animals and man. Part 2) Moscow: Iz d a te ls tv v Academii Nauk SSSR, 499 pp. (In Russian). 90 Taft, Celeste. 1961. The shell-bearing land snails of Ohio. Bull., Ohio Biol. Survey, New Series, Vol. I, no. 3. Publ. by the Ohio State Univ. xi and 108 pp., illus. Thorne, Gerald. 1961. Principles of Nematology. McGraw- H ill Book Co. 553 pp. Travassos, L. 1931. Pesquizas helminthologicas realisadas em Haraburgo, IX. Ensaio monographico da familia Cosmo cercidae. Trav., 1925 (Nematoda). Mem. Inst. Osw. Cruz. 25(3):237-298. 43 pis. Walton, A. C. 1938. The nematoda as parasites of Amphibia. IV. Trans. Amer. Micro. Soc. 57:38-53. biblio. Walton, A. C. 1940. Some nematodes from Tennessee Amphibia. J. Tenn. Acad. Sci. 15:402-405. Walton, A. C. 1941. Notes on some helminths from California Amphibia. Trans. Amer. Micro. Soc. 60:53-57; 1 fig .s b ib lio . Walton, A. C. 1946a. Parasites of Amphibia: Brachycepha- lidae and Hylidae. J. Parsitol. 32 (Sec. 2, suppl.):19. Walton, A. C. 1946b. Parasites of Hylidae, Parts III, IV, and V. Anat. Record 96:591-593. Walton, A. C. 1950a. Parasites of the Brevicepitidae (Amphibia). J. Parasitol. 36 (No. 6, Sect. 2, suppl.): 40. Walton, A. C. 1950b. Parasites of the Amphibia Nematoda. J. Parasitol. 36 (No. 6, Sect. 2, suppl.):40-4l. Wilkie, J. S. 1930. Some parasitic nematodes from Japan. Ann. and Mag. N at. H ist. London. 1 0 (6 ):606-6l4. Yamaguti, S. 1961. The nematodes of vertebrates. Systema Helminthum. Vol. Ill, Parts I and II. Interscience Pub., Inc., New York. 1261 pp. Yorke, W. and P. A. Maplestone. 1926. The nematode parasites of vertebrates. J. & A. Churchill, London. 536 pp. 91 EXPLANATION OP PLATE I Figure 1 . Male tail of Cosmocercoides dukae, lateral view. Figure 2. Male tail of the holotype of Cosmocercoides dukae (Holl, 1928) Wilkie, 1930. Figure 3. Gubernaculum of the holotype of C. dukae, dorsal view. Figure 4. Rosette from the holotype of C. dukae. Figure 5. Male of C. dukae, lateral view. Figure 6. Cephalic region of C. dukae, lateral view. Figure 7. Excretory system of C. dukae, ventral view. Abbreviations: CL - cloaca IR - Intestino-rectal valve EJ - ejaculatory duct PH - phasmid EP - excretory pore SP - spicule GU - gubernaculum TE - testis 92 PLATE I v \ SP' r \ r s r * p . ^ rv r\ ^ 0.5 T £ EXPLANATION OF PLATE I I F ig u re 8. Complex p a p illa (ro s e tte ) of Cosmocercoides dukae, showing central papilla, central microtubercles and outer tubercles. F ig u re 9. Rosette of Cosmocercoides pulcher Wilkie, 1930. F ig u re 10. Rosette of C. pulcher, from cloacal region, lateral view. F ig u re 11. Anterior end of C. dukae, lateral view. F ig u re 12. Rosette of C. dukae, with atypical double p a p illa . F ig u re 13. Gubernaculum of C. dukae from Bufo americanus, Laurel Twp., Hocking Co., Ohio. Dorsal view. F ig u re 14. Gubernaculum of C. dukae from Bufo americanus, Laurel Twp., Hocking Co., Ohio. Dorsal view. F ig u re 15. Gubernaculum of C. dukae from Bufo can or us,- Yosemite National Park. Dorsal view. F ig u re 16. Gubernaculum of C. dukae from Bufo quercicus, Ft. Myers, Florida. Dorsal view. F ig u re 17. Male tail of C. dukae from Scaphiopus holbrooki, Berne Twp., Fairfield Co., Ohio. Lateral view. F ig u re 18. Male tail of C. pulcher from type. Lateral view . PLATE n 95 EXPLANATION OF PLATE I I I F ig u r e 1 9 . Female reproductive tract of Cosmocercoides dukae, vulva and vagina. Lateral view. Figure 20. Tail of female Cosmocercoides pulcher. Lateral view. Figure 21. Tail of female C. dukae. Ventral view. Figure 22. Tail of female C. dukae. Lateral view. Figure 23. Reproductive tract of female C. dukae, one ovary, oviduct and uterus omitted. Figure 24. Gubernaculum and cloacal opening of C. pulcher. Ventral view. Figure 25. Region of vulva of C. pulcher, showing config uration of vagina. “Yentro-lateral view. Figure 26. Cuticle and a micropapilla of the holotype of C. dukae. Figure 27. Cells of the intestine of C. dukae. Abbreviations: Al- ala PH- phasmid MP- micropapilla SR- sem inal re c e p ta c le 0D- oviduct UT- uterus 0V- ovary VA- vagina PA- papilla of anterior VU- vulva cloacal lip 96 PLATE 01 20 •\JA VU, ,MP At PH 2 SR •UT 0V vu 25 24 VA- 26 vUT 27 97 EXPLANATION OF PLATE IV F ig u r e 28 En face view of Cosmocercoides dukae from Bufo americanus, Laurel Twp., Hocking Co., Ohio. Figure 29 En face view of Trionchonema rusticum Kreis, 1932. Figure 30 En face viewof C. dukae, after Ogren, 1953. Figure 31 En face viewof C. dukae, after Anderson, i 960 . Figure 32 En face viewof C. pulcher, after Hsu, 1933. Figure 33 Egg o f C. dukae , single cell. Figure 3^ Egg of C. dukae with two-celled embryo. Figure 35 Egg o f C. dukae w ith three-celled embryo. Figure 36 Egg of C. dukae with four-celled embryo. Figure 37 Egg o f C. dukae with eight-celled embryo. Figure 38 Egg of C. dukae with "tadpole" la rv a . Figure 39 Egg o f C. dukae with well-developed first stage la rva. Abbreviations: AM - amphid DL - dorsal lip LU - lumen PB - polar body 98 plate nr PL LU AM 29 30 28 0.02 O 32 33 0.02 35 36 38