Proceedings of the Helminthological Society of Washington 34(1) 1967

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Volume 34 January \196T Numt>er

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, ALEXA*P>EH I),, AND >O. ^ZÊOBD/QLSEN. "Studies on the Xife 'History of Two Nptoebtylids (Trenaatoda) ___i™_^ ___ _^^l_l_i._. ___ _..; 43 ^ Plicatobothrium cypseluri n. gen., n. sp..XCestoda: 'Pseudophyllidea) from tiie Caribbean Frying Fish, Cypselurus bahiensis (Ranzani,'1842) -i.^w._-,-- ___-~. ______î.^__—^_-^_ 15 , DAVTO J., AND JOHN M. VETTEBUNG.' Cultivation iof ihe Turkey ,Gpccidium, Eimeria meleagrimitis lyzzSr, 1929, in Mammalian Kidney Cell jGultures ^_.^-.....™.-.. ____ .____:._,__.._^.,.j _ 1______„____!; 59 DYER, WILUAM, .G., AND O. WttiFOBD C)LSEN. Biology"-: of Masfophorus / -^nwnidicd (Seurat, 1914) Read and Millemann, 1953 (Nematoda: Spiru- f ridae) with a Description of the Juvenile Stages —^—-^.-ÎL.r.:_-;.', -J,, 98 ^ 3 - (^Coh*in«cdonBocfc Gouer) ' j;f

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^v:-:g-/-5St•'•.•-•;••- -. f _: • ' Copyright © 2011, The Helminthological Society of Washington X: THE HEIMINTHOLOGICM SOCIETVÔF ^WASHINGTd^ < S SOCJE3T meets once a. month irbm October through May for Ithe presentation .and xdi«cîon of .papers m ^ persons ' are irrvited: to attend. _ ~^ : - :-^ ; -^ ' ' ,-. ^'^ "^T--"^--' " '^- -^ "'"- >- v Persons interested in membership m&e^ of "Washington niay obtain application°blar3cs Tfrom the Clprresponding^ Secretary-Treasurer, Miss Edna^ M./Etdirer, Belts-; Aêârajitological Laboratory, Agriculturd Researc^ Center^ Beltsville^ -Maryland 20705. A year's.-jsulbscriptionlto the Proceedings^ is included in, the: i annual dues X'$6.00-). f ;fC 'r :.'^:1'"'!^---^ >/4-^ • /--:-: -{ '•"/ -"-V ;-^ :- v'- "-^:"' -,^ .-J.'""^ %^ OFFICERS OF^HEÔCIETY BELLE Vice President: ^WILLIAM ;B; XteWITT_ v ^> ^ "--;-v: ; Secretary-Treasurer: EDNA M. BUHRER^- '^' HALSEY H. YEGORS ecretary: ALAN C; EIPKIN v - ; ™"i^ V ^ librarian: JUDITH M. HUMPHREY , Jiepresent&iveio tfwWjtshington Acftdemy of^ciences:- AUREL Q. FOSTER ^1965^- ,) DAVID H. Xr^CIGOME, 1967 -5

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_ 'GILBERT F. OTTO, 1969^ " ; -: WILLIAM d- HARGlS, JR., 1971 ^ •>-''' •''••~~~r---'' -'~:,' .• ^- '~;* • '~v ' - ---'- „-.-->-.•.- ~ --••'" -' *'• '-•- "'".'•' •_ -"--•-.'" ^ ^j v- '•'-' •"-'•• (Universi^ of .Maryland -^ <:\ -Virginia Institute^pf-Marine Science Park, Marylarid v~?^: rt. ^_ ^ ;-<^,'

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Experiment Station S. < "";,.. institut~~e o~f "Allergy and' Infectious Diseases^

VOLUME 34 JANUARY 1967 NUMBER 1

Failure of Certain Clams and Oysters to Serve as Intermediate Hosts for Angiostrongylus cantonensis^

STUART E. KNAPP AND JOSEPH E. ALICATA

Several species of invertebrate animals have In the present study, attempts were made been reported as intermediate or as paratenic to determine the following: (a) ability of local hosts for the rat lungworm, Angiostrongylus clams (Venerupis phiUppinarum) and oysters cantonensis. Intermediate hosts which have (Crassostrea virginica) to serve as interme- been found naturally infected include: the diate hosts of A. cantonensis; (b) possible land snails, Bradybaena stmMaris, Opeas javan- natural infection of certain species of clams icum, Macrochlamys resplendens, Achatina and oysters in Hawaii and in the island of fulica, Pupina complanata, and Subulina oc- Ulong, Palau Islands, with third-stage larvae tona; the slugs, Deroceras laeve, Vaginalus of A. cantonensis. These are areas in which plebeius, Veronicella alte, Girasia peguensia, murine angiostrongylosis is endemic. and Microparmarion malayanum; and the am- phibious snail, Pila ampullacea. Naturally in- MATERIALS AND METHODS fected paratenic hosts include: the land pla- Clams collected from Kaneohe Bay, Oahu, narian, Geoplana septemlineata; the freshwater Hawaii, on 31 August 1965, were divided into prawn, Macrobrachiiim sp.; the land crab, 4 groups having 8 clams per group. Each Cardisoma hirtipes; and the coconut crab, group was placed in a glass aquarium contain- Birgiis latro. Also, several land and aquatic ing 5,000 ml of continuously aerated sea water snails and slugs have been reported as ex- with a salinity of 15.1%0 and temperature of perimental hosts (Alicata, 1965). 20 ± 1 C. After 96 hr, the clams in the dif- Of particular interest have been the recent ferent aquaria were exposed to first-stage A. reports (Cheng and Burton, 1965 and Cheng, cantonensis larvae as follows: group 1, 38 1966) that the American oyster, Crassostrea larvae in 0.25 ml of water were injected into virginica, and the soft-shell clam, Mercenaria the mantle of each clam using a hypodermic mercenaria, could serve as intermediate hosts syringe equipped with a 4-inch, 22-gauge of A. cantonensis under experimental condi- needle which was inserted between the valves. tions. This finding could have special signif- The clams remained out of the water for 15 icance especially in some of the Pacific islands min after larval injection; group 2, 3,040 lar- where the rat lungworm exists and clams and vae were placed directly into the aquarium oysters may be eaten raw or imperfectly with the clams; group 3, similar to group 1 cooked. except that 76 larvae were injected into each clam; group 4, same as group 2 except that 1 Contribution from the Department of Veterinary Med- 6,080 larvae were placed in the water. No icine, Oregon State University, Corvallis, and the Parasitol- ogy Laboratory, Department of Animal Sciences, University sand was used for a substrate so the clams of Hawaii, Honolulu. This investigation was supported by Research Grant NB- rested on the glass bottom of their respective 04965-03 of the Institute of Neurological Diseases and aquaria. Blindness, NIH, USPHS. The authors wish to acknowledge the assistance of Mr. Oysters collected from Pearl Harbor, Oahu, Eugene Burke and Mr. Kenji Ego of the Hawaii Fish and Game Commission for assistance in obtaining clams and Hawaii, on 1 February 1966, were divided oysters from the island of Oahu. Mr. Peter Wilson, Fish- into 6 groups having 5 oysters per group. eries Officer of the Trust Territory of the Pacific, supplied the clams from Ulong, Palau Islands. Mr. David Hashimoto Groups 1, 2, and 3 were placed in a glass of the U. S. Fish and Wildlife Service assisted in determining salinity of the sea water used in this study. aquarium containing 5 gallons of continuously

Copyright © 2011, The Helminthological Society of Washington PROCEEDINGS OF THE [VOL. 34, No. 1 aerated sea water with a salinity of 15.3%c preparation or artificial digestive methods 21 and a temperature of 20 ± 1 C. Groups 4, 5, and 25 days later, respectively, failed to show and 6 were placed in another similar aquarium either developing or infective third-stage larvae with a salinity of 10.6%(. After 96 hr, the of the parasite. A few (1 to 2) dead and body of each oyster of groups 1 and 3 was undeveloped first-stage larvae were recovered injected with 0.02 ml normal saline solution after artificial digestion of 3 clams from groups containing approximately 5,000 first-stage lar- 3 and 4 which died 11, 12, and 17 days after vae of A. cantonensis. Each injection was exposure to larval infection. Similarly, 2 live carried out by using a hypodermic syringe and many dead and undeveloped first-stage equipped with a 4-inch, 22-gauge needle larvae were found on press preparation of the which was carefully inserted between the tissue of an oyster (group 1) which died 4 valves. The oysters remained out of the water days after injection with first-stage larvae. for 15 min after larval injection. For oysters of It is concluded that under the conditions of groups 2 and 5, the valves of each were slightly these experiments, neither the clam, V. philip- opened with a knife blade to drain the con- pinarum, nor the oyster, C. virginica, serves as taining sea water, and then with the aid of intermediate host of A. cantonensis. These a fine Pasteur pipette, 0.1 ml of saline solu- data do not confirm the report of Cheng and tion containing approximately 10,000 first- Burton (1965) that C. virginica serves as an stage lungworm larvae was ejected into the experimental host for this parasite. It is be- cavity between the two valves. In this opera- lieved also that further tests are necessary to tion, the oysters remained out of the water for verify the reported claim that M. mercenaria 1 hr before they were replaced in their respec- can be infected with A. cantonensis. No nat- tive glass aquaria. Oysters of groups 3 and 6 ural infection with third-stage larvae of A. were removed into two small glass aquaria, cantonensis was found among oysters or clams each containing 2,000 ml of continuously collected from the island of Oahu, Hawaii, nor aerated sea water (salinity 15.3%0 and 10.6C/CC, from the island of Ulong, Palau Islands. respectively) to which approximately 150,000 The failure either to infect or find natural freshly isolated first-stage rat lungworm larvae infection of local clams and oysters with larvae were added. After two days' exposure to larval of A. cantonensis suggests that these mollusks infection, the two groups of oysters were re- have little importance in the transmission of hu- placed in their former larger aquaria. man angiostrongylosis in the Hawaiian Islands. All of the oysters were examined for larvae of A. cantonensis 25 days after injection or SUMMARY exposure to infection. This was carried out An attempt was made to infect the Hawaiian both by press preparation of parts of the tissues edible clam, Venempis philippinarum, and the of each oyster, and by digesting the remaining American oyster, Crassostrea virginica, with parts of the oysters with the use of the pepsin- first-stage larvae of Angiostrongylus cantonen- HC1 digestion technique. sis. The clams were maintained in glass aquaria Oysters and clams which were collected and containing sea water with a salinity of 15.1%,,. examined for natural infection with larvae of The oysters were maintained in similar aquaria A. cantonensis included the following: 150 containing sea water with a salinity of 15.3%C clams, Venempis philippinarum, from Kaneohe and 10.6^c. These mollusks were exposed to Bay, Oahu, Hawaii; 50 clams, Matra thaanumi, infection by either injecting their tissues with from Ulong, Palau Islands; 100 oysters, Cras- first-stage larvae of A. cantonensis or placing sostrea virginica, from Pearl Harbor, Oahu, the larvae in the water. When these mollusks Hawaii. All these mollusks were examined were examined by press preparation or artificial by use of the pepsin-HCl digestion method. digestive methods, none of them was infected with third-stage larvae of the parasite. A few RESULTS AND DISCUSSION undeveloped and dead first-stage larvae were All the clams and oysters which were either found in the tissues of 3 clams and one injected or exposed to infection with first-stage oyster which died a few days after exposure larvae of A. cantonensis and examined by press to infection.

No evidence of natural infection with larvae meningoencephalitis and other neurological of A. cantonensis was found among 150 clams disorders of man and animals. In Advances in Parasitology, 3: 223-248, Academic Press, (V. philippinarum) and 100 oysters (C. vir- London and New York. ginica) collected from the shores of the island CHENG, T. C. 1966. Perivascular leucocytosis of Oahu, Hawaii, nor among 50 clams collected and other types of cellular reactions in the from the island of Ulong, Palau Islands. oyster Crassostrea virginica experimentally infected with the nematode Angiostrongylus LITERATURE CITED cantonensis. J. Inv. Path. 8: 52-58. CHENG, T. C., AND R. W. BURTON. 1965. The ALICATA, J. E. 1965. Biology and distribution American oyster and clam as experimental of the rat lungworm, Angiostrongylus canton- intermediate hosts of Angiostrongylus cantonensis, and its relationship to eosinophilic ensis. J. Parasit. 51: 296.

Myzotrema cyclepti gen. n., sp. n. (Trematoda: Monogenea) from Gills of Cycleptus elongatus (LeSueur) from Alabama1

WlLMER A. ROGERS-

The species described in this paper was off from body proper by stout peduncle, with collected as part of a survey of fish parasites two pairs of nearly similar anchors, each pair being conducted by the Southeastern Coopera- supported by a nonarticulate transverse bar; tive Fish Parasite and Disease Project of the 14 marginal booklets present. Pharynx large, Agricultural Experiment Station, Auburn Uni- heavily muscularized, perfectly round in cross- versity. This species was collected using the section. Intestinal crura simple, united pos- 1:4,000 formalin field-collecting method de- teriorly. Testis and ovary equatorial, over- scribed by Rogers (1966). Specimens were lapping. Vas deferens looped around left in- measured according to the procedure given by testinal crus, seminal vesicle formed by dila- Mizelle and Klucka (1953). Measurements tion of vas deferens. Two prostatic reservoirs are expressed in microns and were made from present. Cirrus a U-shaped tube with complex specimens mounted in glycerin jelly or per- accessory piece articulated to base. Ovary mount. Details of internal anatomy were de- looping around right intestinal crus. Vagina termined from hematoxylin-stained specimens. present, opening dextroventrally; submedian or Illustrations were prepared with the aid of a submarginal. Vitellaria coextensive with intes- camera lucida. The keys to the genera of tine. Parasitic on fresh water fish. Ancyrocephalinae by Mizelle and Price (1964) and Yamaguti (1963) were useful in deter- TYPE SPECIES: Myzotrema cyclepti sp. n. mining the status of the present species. TYPE HOST: Blue sucker, Cycleptus elon- Myzotrema gen. n. gatus (LeSueur). LOCALITY: Tombigbee River, Pickens Coun- GENERIC DIAGNOSIS: Dactylogyridae, Ancy- ty, Alabama. rocephalinae: Body large, elongate, with two REMARK: Myzotrema gen. n. is most closely pairs of eyespots, head organs poorly de- related to Pseudomurraytrema Bychowsky, veloped or lacking. Opisthohaptor well set 1957 (nee Pseudomurraytrema Yamaguti, 1958) as shown by the structure of the copulatory 1 Supported by the Southeastern Cooperative Fish Para- site and Disease Project. complex and the reproductive system. It is - Agricultural Experiment Station, Auburn University, Auburn, Alabama. readily separated from Pseudomurraytrema by

Copyright © 2011, The Helminthological Society of Washington PROCEEDINGS OF THE [VOL. 34, No. 1 having two instead of three haptoral bars. The 8). The median pair is situated immediately copulatory complex of Anoncohaptor Mueller, posterior to the ventral bar which according to 1938 (Mueller, 1938, plate 1, fig. 5) is also Mizelle and Crane (1964) would be pair no. 5. similar to that of the present genus, but Anon- Pairs 1, 2, and 3 would then be located ven- cohaptor has no haptoral anchors or bars. Both trally while pairs 4, 6, and 7 would be located Urocleidiis Mueller, 1934, and Cleidodisciis clorsally. Testis and ovary located slightly an- Mueller, 1934, have two pairs of anchors that terior to midlength; overlapping. Testis glo- are each supported by a transverse non-artic- bose, located sinistrally in loop of ovary; vas ulated bar (Yamaguti, 1963), but in neither deferens looping around left limb of intestine genus does the vas deferens or ovary loop and running anteriorly, expands into seminal around the intestinal limbs as in Myzotrema. vesicle just posterior to cirrus. Copulatory complex composed of cirrus (Fig. 1) and ac- Myzotrema cijclepti gen. n., sp. n. cessory piece (Fig. 2). Cirrus a U-shaped (Figs. 1-8) tapering tube with an angular projection about DESCRIPTION: A large form with elongate midlength, with a complex basally articulating body and thin cuticle that may be finely stri- accessory piece. Two prostatic reservoirs ated. Length 1,885 (1,650 to 2,200), greatest empty into cirrus, one about twice as long as width 301 (185 to 370) about midlength. Four the other. Length of cirrus 50 (48 to 52); eyespots present, approximately same size, pos- length of accessory piece 51 (47 to 55). terior pair sometimes larger, members of pos- Ovary vase-shaped with elongate neck folding terior pair always farther apart. Head without over testis and looping around right intestinal lobes, head organs undeveloped or lacking. limb. Vagina opening ventrally, submarginal Pharynx perfectly round in cross-section, heav- or submedian; with a long coiled tube leading ily muscularized, transverse diameter 131 (115 into a large pear-shaped seminal receptacle. to 150). Intestinal limbs simple, uniting pos- Vitellaria coextensive with intestine, confluent teriorly. Peduncle long and stout; haptor obo- over most of anterior two-thirds of body pos- vate in outline with smooth margins, broader terior to pharynx. than long, length 147 (110 to 210), width HOST AND LOCALITY: Blue sucker, Cycleptus 180 (170 to 210). One pair of dorsal and one pair of ventral anchors, similar in size and elongatus (LeSueur), Tombigbee River, Pick- shape, each pair supported by a transverse bar. ens County, Alabama. Each anchor composed of (1) a broad base SPECIMENS STUDIED: Eight. without differentiated roots, (2) a solid short TYPE SPECIMENS: Type, USNM Helm. Coll. shaft grading into (3) a solid, short finely No. 61248; 1 paratype, USNM Helm. Coll. attenuated point. Anchor wings not observed. No. 61249; paratypes in author's collection. Ventral anchors (Fig. 4) often smaller than REMARKS: Myzotrema cijclepti is the only dorsal anchors; length 59 (57 to 64), width described species in this genus. The nature of of base 36 (35 to 38). Dorsal anchor (Fig. 5) the copulatory complex, vagina, ovary, and length 59 (53 to 64), width of base 38 (34 other reproductive structures is very similar to to 40). Two transverse nonarticulate bars those observed in Pseudomurraytrema species present; ventral bar (Fig. 6) with prominent (Rogers, 1966); P. copulatum (Mueller) and posteriorly projecting arch, dorsal bar (Fig. 7) Anoncohaptor anomaliis Mueller (Mueller, approximately straight but with an anteriorly 1938). directed hump at midlength. Length of ventral bar 73 (72 to 75); length of dorsal bar 89 ACKNOWLEDGMENTS (84 to 96). Marginal hooks 14 in number, similar in size and shape (Fig. 3); small with- Thanks are extended to William Smith- out inflated bases, length 16 (15 to 17). One Vaniz, Division of Fisheries, Smithsonian In- hook pair is located ventrally in center of hap- stitution, Washington, D. C., and to Walter tor while 3 pairs are located ventrally at an- Tatum and other members of the Alabama terior edge of haptor and 3 pairs are located Department of Conservation for assistance in doisally in anterior one-third of haptor (Fig. collecting the fish hosts.

Figs. 1-8. Myzotrema cyclepti gen. n., sp. n. 1. Cirrus; 2. accessory piece; 3. marginal hooks; 4. ventral anchor; 5. dorsal anchor; 6. ventral bar; 7. dorsal bar; 8. entire worm, ventral aspect.

LITERATURE CITED of Micropterus salmoides (Lacepede) from a MIZELLE, J. D., AND A. R. KujCKA. 1953. Stud- California pond. Trans. Amer. Micr. Soc. 83: ies on monogenetic trematodes, XIV. Dacty- 343-348. logyridae from Wisconsin fishes. Amer. Midi. MUELLER, J. F. 1938. Additional species of Nat. 49: 720-733. North American Gyrodactyloidea (Trematoda). Amer. Midi. Nat. 19: 220-235. , AND C. E. PRICE. 1964. Studies on monogenetic trematodes, XXV. Six new spe- ROGERS, W. A. 1966. Three new species of cies of Ancyrocephalinae from the gills of Pseudomurraytrema (Trematoda: Monogenea) Zanclus canescens (L.) with a key to the from gills of Catostomid fishes. J. Parasit. genera of Ancyrocephalinae. ]. Parasit. 50: 52: 462-465. 81-89. YAMAGUTI, S. 1963. Systema Helminthum Vol- , AND J. W. CRANE. 1964. Studies on ume IV. Monogenea and Aspidocotylea. In- monogenetic trematodes, XXIII. Gill parasites terscience Publishers. New York, London.

A Review of the Genus Pseudocella Filipjev, 1927 (Nematocla: Leptosomaticlae) with a Description of Pseudocella triaulolaimus n. sp.

W. DUANE HOPE1

The genus Thoracostoma Marion, 1870 was terior margin of the cephalic capsule, and separated into the subgenera Thoracostoma and ocelli absent. Synonchoides received a single Pseudocella by Filipjev (1927), the former species, T. galathea (Wieser, 1956) which also subgenus to receive those species with lens- resembles species of Pseudocella, but which bearing ocelli, symmetrical, but irregularly possesses a cephalic capsule reduced to a ring curved spicula, and gubernacula with apoph- with a slightly notched posterior edge, no yses directed dorsally and parallel with the anterior lobes, with amphids completely pos- spicula, and the latter subgenus to receive terior to the lobes and a buccal armature re- those species with ocellar pigment spots lack- sembling that of Synonchus Cobb, 1893. From ing a lens, spicula often asymmetrical but uni- the two male specimens of T. galathea, Wieser formly curved, and gubemacula with paired was unable to determine if the gubemaculum apophyses directed posteriorly at right angles possessed caudally directed apophyses. to the spicula. Later, Filipjev (1946) raised Platonova (1962) ranked Thoracostoma and both subgenera to generic rank. Pseudocella as genera and assigned T. kreisi Wieser (1953) continued to treat Filipjev's and T. fttipjevi to Pseudocella without refer- taxa as subgenera of the genus Thoracostoma ence to Wieser's subgenera. and later added two additional subgenera, The differences between Thoracostoma and Corythostoma and Synonchoides (Wieser, Pseudocella have remained much the same 1956). Corythostoma was proposed to receive even though new species have been added to T. kreisi Wieser, 1953 and T. filipjev Kreis, both groups. A notable exception, as pointed 1928 which resemble species of Pseudocella, out by Wieser (1956), is that species of Thora- but which possess a cephalic capsule with costoma have been described which do not weakly developed posterior lobes separated possess ocellar pigment spots or lenses (Wieser, by notches instead of incisures and which lack 1956; Allgen, 1951; Schuurmans Stekhoven incisions or fenestrae, with amphids totally, or and Mawson, 1955). Consequently, emphasis with their greatest portion, behind the pos- has shifted from differences in the ocelli to differences in the spicula and gubemacula. 1 Department of Invertebrate Zoology, Museum of Nat- ural History, Washington, D. C. The fact remains, however, that there are

20 JJ

Fig. 1. Pseudocella triaulolaimus n. sp. Head of holotype in lateral view. Dorsal and subventral interradial teeth not illustrated. relatively distinct and, at present, consistent be provisionally considered a subgenus of morphological differences between these two Pseudocella. A more satisfactory arrangement groups and, for these reasons, their generic must be based on additional studies of the status is herein retained. stomatal armature, spicula, and gubernaculum Species of Corythostoma differ from those of P. (Synonchoides) galathea (new combina- of Thoracostoma and Pseudocella in the struc- tion). The taxonomic status of this species ture of the cephalic capsule, as mentioned might also be clarified by a detailed study of above, and for this reason, retaining Cory- the stomatal armature in species of the genus thostoma as a subgenus can be justified, pend- Synonchus. ing further study. On the other hand, it is All species of the genus Pseudocella not here difficult to justify raising Corythostoma to the assigned to the subgenera Corythostoma and generic level since the males of this group, Synonchoides are regarded as members of the that have been adequately studied, have spic- nominate subgenus Pseudocella, which has the ula and gubernacula identical to those of posterior lobes of the cephalic capsule sep- Pseudocella. Therefore, the subgenus Cory- arated by notches; a claviform piece present; thostoma is here transferred from Thoraco- amphids completely surrounded by lobes of stoma to the genus Pseudocella. the cephalic capsule; ocelli absent; spicula of It is also here proposed that Synonchoides uniform diameter and regularly curved; and

Fig. 2. Pseudocella triaulolaimus n. sp. Photomicrographs of cross-sections through stoma. Sections A through D approximately 5, 10, 15, and 30 /j., respectively, from anterior extremity. Anterior dorsal interradial tooth (ad); teeth of radii (t); subventral interradial teeth (s); posterior dorsal interradial tooth (pd). gubemacula with caudally directed apophyses in sea water and mounted in glycerine or (Wieser, 1956). CMC10*. The latter was used principally for males, for the specimens are rendered more Pseudocella (Corythostoma) transparent in this mounting medium and a triaulolaimus n. sp. detailed study of the stomatal structure and Sixteen male, three female, and three juve- male genital apparatus is made possible. The nile specimens of Pseudocella triaulolaimus head of one male was mounted en face in n. sp. were collected in February, 1963 from glycerine jelly and one male and one female sandy sediment held by holdfasts of Egregia were embedded in polyethylene glycol and menziesii (Turner). The holdfasts were at- sectioned at 5 />,. The sections were stained tached to rocks in the intertidal zone at Dillon with hematoxylin at pH 2.4 by the method of Beach, California. The nematodes were re- Craig and Wilson (1937). laxed for 30 minutes in sea water saturated with carbon dioxide, fixed in 10% formalin :': Product of Turtox Biological Supply House Inc.

A , 100 Lt

B , 100 U

C.D, 50 LI

Fig. 3. Pseudocella triaulolaimus n. sp. (A) Tail of allotype (female). (B) Tail of holotype (male). (C) Left (with distal hook) and right spicula of male paratype. (D) Gubernaculum of male paratype.

MEASUREMENTS approximately 30 ^ posterior to teeth (Fig. HOLOTYPE (Male): L = 16.80 mm; a = 2D). Further posteriorly, distal ends of radii 74.7; b = 8.27; c = 69.13; T = 45%. convergent, walls thin but with muscle attach- ALLOTYPE (Female): L = 19.72 mm; a — ment points and muscles concentered. Esophagus muscular and gradually increas- 88.4; b = 8.80; c = 93.0; V = <24-G%> 62.9% (22.6%). ing in diameter towards its posterior end. PARATYPES (13 males): L = 14.53-20.89 Nerve ring located approximately % the length mm (18.75); a = 51.5-83,5 (72.23); b = 7.27- of the esophagus from the anterior end. Re- 13.47 (9.19); c = 60.04-88.60 (73.09); T = nette cell and ocellar pigment spots absent. 45.0%-71.6% (60.9%). MALES: Longitudinal series of subventral PARATYPE (1 female): L= 18.09 mm; a = setae extending from near mid-region of tail to posterior subventral supplements; 16—22 in 58.7; b = 8.89; c = 88.7; V = <14-8%> 64% (18.0%)_ right subventral series, 17-21 in left subven- PARATYPES (3 juveniles): L = 12.06-13.96 tral series; setae 15-25 ^ long. Four to six (13.15); a = 50.7-72.7 (62,6); b = 6.03-8.31 setae distributed irregularly around caudal (7.46); c = 58.1-73.9 (6.77). gland pore. Heavily cuticularized, hemispher- DESCRIPTION: Body long, slender, and taper- ical copulatory supplements in one subventral ing anteriorly from base of esophagus; pos- row on each side of body anterior to anus; four terior extremity tapering from short distance to six supplements on right side, four to seven anterior to cloaca. Integument without trans- on the left side; position of supplements rela- verse or longitudinal striations; cuticle 10.5- tive to those of the opposite side either even 6.1 (8.69) p. thick at mid-body level. Six or staggered; each supplement bearing a papillae on lips followed by circle of ten minute apical papilla. cephalic setae, the latter, 11.0 to 17.5 ^ long. Reproductive system cliorchic, testes opposed Cephalic capsule with well-developed pos- and outstretched. Spicula (Fig. 3C) rather terior lobes; sublateral lobes only partially uniformly arcuate and comprised of three enclosing amphids; lobes separated by deep longitudinal ribs; proximal end of spicula de- notches and lacunae absent. Anterior lobes void of dilated capitulum; distal end of left short; claviform piece absent. Major portion spiculum thickened, with a pointed posterior of cephalic capsule separated from integumen- process. Right spicula 292.6-319.5 /«,, left spicula 270.7-324.4 p. long. Corpus of gubemac- tary cuticle by alveolar substance (tissue?). Amphids circular, % width of corresponding ulum (Fig. 3D) with thin-walled processes embracing spicula laterally and with heavily head diameter and situated just posterior to cuticularized apophyses extending at right lateral cephalic setae. angles to spicula in a dorso-caudal direction; Cervical setae between cephalic capsule and apophyses fused at their bases by a medial nerve ring in dorsal, lateral, and ventral longi- triangular thickening. Paired trumpet-shaped tudinal rows. These setae long (11.0 to structures (Fig. 3D), ventro-lateral to distal 17.0 /A), numerous and in clusters near pos- end of spicula, apparently fused laterally with terior margin of cephalic capsule, becoming gubemaculum; these structures with a small sparse and shorter (3.0 to 5.0 ^} at level of nerve ring. Setae posterior to nerve ring 2.0 pore near their distal end. to 3.0 p. long and sparsely distributed. Posterior end of body strongly curved Stoma triradiate in cross-section with 12 toward venter. Copulatory muscles well de- teeth, two at distal end of each radius (Fig. veloped in approximately the posterior 11% 2A and B; t) and two on each of three inter- of the body length. Tail conical with terminal radial facets; sub ventral interradial teeth jux- gland pore; caudal glands convoluted, not tapose (Fig. 2B; s) and dorsal interradial extending anterior to gubemaculum. teeth tandem; anterior dorsal tooth 1.0 ^ long FEMALES: Vulva without distinctly pro- (Fig. 2A; ad), posterior dorsal tooth 3.5 ^ truding lips. One to three rounded cuticular long (Fig. 2C; pd); all teeth directed antero- elevations anterior and posterior to vulva on medially. Distal ends of radii tuboid and walls ventromedial body surface (Fig. 4); setae not of radii greatly thickened for a distance of present on these elevations. Five to six hypo-

Fig. 4. Pseudocella triaulolaimus n. sp. Allotype (female).

Copyright © 2011, The Helminthological Society of Washington 12 PROCEEDINGS OF THE [VOL. 34, No. 1 dermal gland cells in longitudinal series in Pseudocella triaulolaimus n. sp. can be dis- each lateral hypodermal chord at level of tinguished from all species of Pseudocella by vulva; gland cells opening to the exterior by the tuboid radii and thickened walls in the minute pores on ventral margin of each lateral posterior portion of the stoma and by the chord. greater number of stomatal teeth. Reproductive system amphidelphic and reflexed. Ova with chorion 560-653 (601) p LITERATURE CITED long and 127-147 (136.5) ^ wide. Tail (Fig. ALLGEN, C. 1951. Papers from Dr. Th. Morton- 3A) conical, terminus bluntly rounded. sen's Pacific Expedition 1914-1916. LXXVI. HOLOTYPE (Male): United States National Pacific free-living marine nematodes. Viden- Museum catalogue number 33631. skabelige mecldelelser Dansk Naturlristorik ALLOTYPE (Female): United States Na- forening, Copenhagen, p. 113. tional Museum catalogue number 33632. CRAIG, R., AND CHARLES WILSON. 1937. The PARATYPES (Males, female, and juveniles): use of buffered solutions in staining; theory and practice. Stain Technology 12 ( 3): 99- United States National Museum catalogue 109. numbers 33633-33650. FILIPJEV, I. N. 1927. Les nematodes libres des TYPE HABITAT: Sediment held by the hold- mers septentrionales apartenant a la famille fasts of Egregia menziesii (Turner) attached des Enoplidae. Archiv fur Naturgeschichte to intertidal rocks. 91(6): 1-126. TYPE LOCALITY: Dillon Beach, California. . 1946. Free-living nematodes from the REMARKS: Pseudocella triaulolaimus n. sp. northern polar basin. Study of expedition of Glavsevmorputy on the ice cutter ship Sedov has been assigned to this genus because of the 1937-1940. (In Russian.) 3: 158-184. presence of a cephalic capsule, a relatively KREIS, H. A. 1928. Die freilebenden marinen short, bluntly rounded tail, uniformly curved Nematoden der Spitzbergen-Expedition von spicula and gubernacula with caudally directed F. Roemer und F. Schaudinn im Jahre 1898. apophyses; to the subgenus Corythostoma be- Mitteilungen aus dem Zoology Museum in cause the lobes of the cephalic capsule are Berlin. 14(1): 131-197. reduced in length and separated by notches PLATONOVA, T. A. 1962. New species of nem- only, neither incisions or fenestrae are present atodes of the genus Pseudocella Filipjev from the Kurile Islands and Southern Sakhalin (In and amphids have at least half their length Russian). Studies of the Far Eastern Seas of situated posterior to the caudal margin of the the USSR. USSR Academy of Sciences, Mos- lobes. cow-Leningrad 8: 200-218. It might be argued that P. triaulolaimus SCHUURMANS STEKHOVEN, J. H., AND P. M. MAW- belongs in the subgenus Synonchoides because SON. 1955. On some free-living marine of the multiple teeth in the stoma. It is not nematodes from Kerguelen Island. I. Hel- assigned to this subgenus since the cephalic minth. 29(1/2): 87-104. capsule is not reduced to a narrow band and WIESER, W. 1953. Free-living marine nema- because it is difficult at this time to appraise todes I. Enoploidea. Reports of the Lund University Chile Expedition 1948-1949. 10: the apparent similarities of the stomatal arma- 1-155. ture without knowing the exact structure and . 1956. Some free-living marine nema- location of teeth present in P. (S.) galathea. todes. Galathea Report 2: 243-253.

Incidence of Bovine Coccidia in Western Oregon1

PETER A. NYBERG, DONALD H. HELFER, AND STUART E. KNAPP

Eleven or more species of coccidia of the there, and because it has climatic conditions genus Eimeria are currently recognized as oc- representative of other Pacific Northwest dairy curring in bovine animals. At least 9 species areas. Practitioners were asked to randomly are known to occur in this country, their inci- collect fecal samples from two-week to one- dence having been determined through field year-old dairy calves between August and surveys of cattle conducted in various parts November, 1965. They were also asked to of the United States, particularly the South- record the animal's sex, age, and fecal con- eastern, North-Central, and Western states sistency for each sample submitted. (Christensen, 1941; Davis, et al., 1955; Hasche The approximate number of oocysts per and Todd, 1959; Fitzgerald, 1962). These gram of feces was determined using a modified species include E. bovis, E. zurni, E. ellip- McMaster method (Whitlock, 1948). After soidalis, E. auburnensis, E. cylindrica, E. sub- weighing, contents of each tared sample bottle spherica, E. canadensis, E. bukidnonensis, and were emptied into a beaker and water added E. alabamensis. Two species, E. bovis and E. to make a total volume of 100 ml. Following zurni, are consistent pathogens and cause acute thorough mixing, a 1.0 ml aliquot (1/100 of coccidiosis (Hammond, et al., 1944; Boughton, original sample) was combined with 2.0 ml 1945; Davis and Bowman, 1951). Eimeria Sheather's sugar solution. After vigorous shak- ellipsoidalis and E. alabamensis may produce ing, the oocysts in 1/20 (0.15 ml) of this no observable symptoms unless large numbers 3.0 ml mixture were counted, using a Mc- of sporulated oocysts are ingested or unless Master counting chamber (representing 1/2,000 the calf is weakened from other predisposing of the fecal material in the original sample). conditions (Davis and Bowman, 1956). Ei- meria. cylindrica was reported by Wilson (1931) RESULTS AND DISCUSSION (as cited by Levine, 1961, p. 172), to be moderately pathogenic while E. auburnensis, E. Incidence and identification of species bukidnonensis, and E. canadensis have a rela- Of a total of 86 fecal samples examined, 62 tively low degree of pathogenicity (Ham- (72%) contained oocysts of cither a single or mond, et al., 1961; Levine, 1961). The patho- a mixture of species. From these samples, 8 genesis of E. subspherica is unknown. species of coccidia were identified (Christen- Although bovine coccidia have been found sen and Porter, 1939; Christensen, 1941; Ny- in Pacific Northwest livestock, no incidence berg and Hammond, 1965). In order of their report for the various species has been made. decreasing frequency, they were: Eimeria It was for this reason that the present study bovis, E. ellipsoidalis, E. zurni, E. auburnensis, was undertaken. E. cylindrica, E. subspherica, E. bukidnonensis, and E. alabamensis. MATERIALS AND METHODS Table 1 shows the majority of positive sam- Tared sample bottles containing 2.5% po- ples contained mixtures of the various species; tassium dichromate were distributed among however, E. bovis, E. ellipsoidalis, E. auburn- three veterinary offices in Tillamook County, ensis, E. zurni, E. cylindrica, and E. bukid- Oregon. This geographic location was selected nonensis were found occurring as single infec- because of the concentrated dairy industry tions, representing 28% of the total samples. Discharge of oocysts The number of oocysts per gram of feces ranged from 500 to over 10 million for the various species present. The range for the

TABLE 1. Incidence of bovine coccidia in 86 5,000 oocysts or more per gram of feces, how- dairy calves from Western Oregon. ever, only 25% were scouring. Species of coccidia Number of occurrences: LITERATURE CITED Total Single Mixed E. bovis 53 (62%) 18 (21%) 35 (41%) BOUGHTON, D. C. 1945. Bovine coccidiosis: E. ellipsoidalis 28 ( 33% ) 2 ( 2%) 26 (31%) from carrier to clinical case. North Am. Vet. E. zurni 20 (23%) 1 ( 1%) 19 ( 22% ) E. anbiirnensis 12 (14%) 2 ( 2%) 10 ( 12% ) 26: 147-153. E. cylindrical 9 (10%) 1 ( 1%) 8 ( 9%) CHRISTENSEN, J. F. 1941. The oocysts of coc- E. subspherica 7 ( 8%) 0 ( 0%) 7 ( 8%) cidia from domestic cattle in Alabama (USA) E. bnkidnonensis 5 ( 6%) 1 ( 1%) 4 ( 5%) E. alabamensis 1 ( 1%) 0 ( 0%) 1 ( 1%) with descriptions of two new species. J. Parasit 27: 203-220. , AND D. A. PORTER. 1939. A new coc- two most pathogenic species (E. bovis and cidium from, cattle, with observations on its life history. Proc. Helminthol. Soc. Wash. E. zurni) was from 425 to 2 million, and 370 6: 45-48. to 8 million, respectively. A previous report DAVIS, L. R., D. C. BOUGHTON, AND G. W. BOW- (Horton-Smith, 1958), stated that a discharge MAN. 1955. Biology and pathogenicity of rate in excess of 5,000 oocysts per gram is Eimeria alabamensis Christensen, 1941, an indicative of clinical coccidiosis. We found intranuclear coccidium of cattle. Am. 1. Vet. 36 of the 62 positive animals (58%) passing Res. 16: 274-281. 5,000 or more oocysts of E. bovis or E. zurni, —, AND G. W. BOWMAN. 1951. Coccidiosis per gram of feces. Of these, 21 (59%) had in cattle. Proc. U. S. Livestock Sanit. Assoc. 55th Ann. Meet., p. 39-50. normal stools, 9 (25%) were loose or scouring, , AND . 1956. Bovine coccidiosis. and no history was given for 6 animals. The Yearbook of Agriculture. U. S. Govern- Twenty of 22 negative animals, for which ment Printing Office, Washington, D. C. p. the age, sex, and consistency of feces was 314-316. known, were 4 months old or older. Perhaps FITZGERALD, P. R. 1962. Coccidia in Hereford this finding is related to the immunity ob- calves on summer and winter ranges and in served in older animals that have been pre- feedlots in Utah. J. Parasit. 48: 347-351. viously exposed to bovine coccidia. HAMMOND, D. M., W. N. CLARK, AND M. L. MINER. 1961. Endogenous phase of the life SUMMARY cycle of Eimeria auburensis in calves. J. Parasit. 47: 591-596. Sixty-two (72%) of 86 fecal samples col- -, L. R. DAVIS, AND G. W. BOWMAN. 1944. lected randomly from dairy calves in Tillamook Experimental infections with Eimeria bovis in County, Oregon, contained oocysts of the calves. Am. J. Vet. Res. 5: 303-311. genus Eimeria. Eight species were identified. HASCHE, M. R., AND A. C. TODD. 1959. Prev- Listed in order of their decreasing frequency, alence of bovine coccidia in Wisconsin. J. Am. they were: Eimeria bovis (62%), E. ellip- Vet. Med. Assoc. 134: 449-451. soidalis (33%), E. zurni (23%), E. aubiirnen- HORTON-SMITH, C. 1958. Coccidiosis in domes- sis (14%), E. cylindrica (10%), E. subspherica tic mammals. Vet. Rec. 70: 256-262. (8%), E. bukidnonensis (6%), and E. ala- LEVINE, N. D. 1961. Protozoan parasites of domestic animals and of man. Burgess Pub- bamensis (1%). Twenty-eight per cent of the lishing Co., Minneapolis, Minn., 412 p. 86 samples contained single species, of which NYBERG, P. A., AND D. M. HAMMOND. 1965. E. bovis comprised 21%. All species except Description of the sporulated oocysts and E. subspherica and E. alabamensis occurred sporozoites of four species of bovine coccidia. singly. J. Parasit. 51: 669-673. Oocyst discharge, involving all samples, WHITLOCK, H. V. 1948. Some modifications of ranged from 500 to over 10 million per gram the McMaster helminth egg-counting tech- of feces. Fifty-eight per cent of the animals nique and apparatus. J. Counc. Scient. Inclust. harboring E. bovis and .E. zurni were passing Res., Australia, 21: 177-180.

Plicatobothriurn cypseluri n. gen., n. sp. (Cestoda: Pseudophyllidea) from the Caribbean Flying Fish, Cypselurus bahiensis (Raiizani, 1842)

R. M. CABLE AND MARY B. MICHAELIS'

The adult helminths found in 185 species like spines. Neck very short if present; strobila of marine teleosts examined in Curacao and acraspedote, distinctly segmented only at inter- Jamaica in 1961 included 178 species of vals. Genital pore dorsal, median, posterior to digenetic trematodes and seven of acantho- midlevel of proglottid. Uterine pore midven- cephala, but only two species of cestodes were tral, inconspicuous, at posterior end of large, recovered. Both were pseuclophyllideans. Al- Y-shaped uterine sac with thin wall. Ovary at though the scolex was not found for a speci- extreme posterior end of proglottid, flanked by men from the houndfish, Strongylura timucu arms of uterine sac in following proglottid. (Walbaum, 1792), examined in Jamaica, the Uterine duct long, extensively coiled in zone strobila identified the cestode as a species of between ovary and genital pore, then extend- Ptychobothrium Loennberg, 1889, possibly P. ing anteriorly to enter stem of uterine sac belones (Dujardin, 1845). Cestodes referred dorsolaterally at a distance from its anterior to that species have been reported from hound- end. Vitelline follicles densest laterally but fishes in widely separated localities. Four com- encompass medullary parenchyma except in plete specimens of the second cestode were vicinity of reproductive pores; follicles between removed from the intestine of flying fish in conspicuous layers of longitudinal muscle fibers Curacao, fixed in corrosive sublimate-acetic that are not grouped into distinct bundles. acid, and preserved for study later. Three Testes in lateral regions of medullary paren- were prepared as whole mounts stained with chyma. Cirrus sac without external prostatic Harris' hematoxylin or Semichon's carmine fol- cells. Eggs thin-shelled non-operculate. Adults lowed by indulin to bring out surface features. in intestine of marine fish, life history un- The scolex of the fourth specimen was used known. Type and only species: for cross-sections and portions of the strobila for serial sections cut in the transverse and Plicatobothrium cypseluri n. gen., n. sp. frontal planes. Drawings were made by micro- (Figures 1-7) projection and measurements are in millimeters. DESCRIPTION (based on four mature speci- Because features of the species from flying mens) : with the characters of the genus. fish exclude it from any existing genus, a new Scolex disproportionately large in comparison one is erected to receive it and is characterized with strobila, triangular from lateral aspect of as follows: fixed specimens, 2.5—2.9 long, 1.3—1.8 in maxi- Plicatobothrium n. gen. mum width; its shape in living specimens continuously changed by movements of extremely Order Pseudophyllidea, Family Ptychoboth- active, muscular bothria which overhang junc- riidae. Scolex basically H-shaped in cross- tion of strobila and scolex. Strobila about 50.0 section, triangular to fan-shaped from lateral long; proglottids wider than long until gravid, aspect, without apical organ or hooks. Bothria then becoming much longer than wide. Pri- two, deep, open their entire length, not con- mordia of reproductive organs evident a short nected at tip of scolex; their walls extensively distance from scolex. Uterine sacs develop in convoluted and folded, lined with minute, hair- immature proglottids, first the more dorsal stem as a spherical cavity, then sacculations

1 Department of Biological Sciences, Purdue University, that become the arms; sacs Y-shaped with Lafayette, Indiana. Specimens serving as the basis of this paper were collected in 1961 as part of an investigation short arms in extended proglottids, cordiform then supported by Grant G-14691, National Science Foun- when distended in contracted ones; uterine dation. Laboratory facilities were kindly extended by the Caribbean Marine Biological Institute, Curacao, N.A., and pore median, slightly posterior to genital pore by the Marine Biological Laboratory, University College of the West Indies, Jamaica, W.I. on opposite side of proglottid. Vitelline reser-

Copyright © 2011, The Helminthological Society of Washington JANUARY, 1967] HELMINTHOLOGICAL SOCIETY 17 voir dorsal, conspicuous, always tilted to side Features of Plicatobothrium cypseluri place on which uterine duct enters uterine sac: it in the Family Ptychobothriidae Liihe, 1902, usually on left but irregularly alternating; cir- as recognized by Wardle and McLeod (1952) rus sac always tilted from median line to and Yamaguti (1959). It seems closest to opposite side. Ovary median, triangular in Ptychobothrium belones, the only described outline but appearing broadly chevron- to V- species of Ptychobothrium which, like Plica- shaped in whole mounts, depending on con- tobothrium, has a scolex that bears only two traction, because of greater thickness at sides deep bothria, open their entire length and not where lobules are dorsal to thinner connecting connected anteriorly. Further indication of portion. Oocytes collect in that portion; ovi- their affinity is suggested by the fact that duct begins there with oocapt, followed by synentognath fishes serve as hosts of both enlarged, muscular segment, joined clistally by species. In other respects, however, they show small spherical enlargement at end of vagina. differences that justify their allocation to sep- From that junction, oviduct extends as a nar- arate genera. Unlike Plicatobothrium, Ptycho- row tube to end of vitelline reservoir where it bothrium has the genital pore anterior to the turns, receives vitelline duct, and joins ootype midlevel of the proglottid, and a uterine duct well to side of median line. At ootype, uterine that extends from the female complex in a duct begins as a thin-walled tube which forms wide curve to the right or left, to cross the several loops, then becomes heavily muscular anterior end of the proglottid and join a small, and much convoluted before extending ante- spherical uterine sac which opens at a con- riorly to enter uterine sac. From its enlarge- spicuous pore far to one side of the median ment at junction with oviduct, vagina also line. Moreover, eggs develop embryos with begins as a sinous, thin-walled tube, often with hooks and are stored largely if not entirely in several large dilatations, then becomes heavily the uterine duct which consequently becomes muscular before extending anteriorly to open distended with eggs and forms convolutions into genital atrium; a short distance from that that fill most of the medullary parenchyma in opening, vagina often with conspicuous en- gravid segments. largement sometimes containing oocytes as Although Ptychobothrium and Plicatoboth- well as sperms. Testes scattered in lateral rium are at present monotypic genera, it seems medullary parenchyma from ovary to bifurca- likely that further and more precise studies tion of uterine sac. Cirrus sac oval to pyriform, will reveal additional species. During his measuring 0.128-0.155 by 0.10-0.122 in ma- studies in the Galapagos Islands, Manter (per- ture proglottids, its length increasing to 0.162- sonal communication) found in a flying fish 0.182 in terminal portion of strobila in which a scolex resembling that of P. cypseluri but uterine sacs are dilated but almost empty. of a somewhat different shape. Even greater Eggs oval, non-collapsed ones in whole mounts differences have been described for scoleces 0.045-0.048 long, 0.031-0.037 wide; none de- of cestodes that have been obtained from velop in uterus sufficiently to show larval houndfishes and identified as P. belones. hooks. HOST: Cypselurus bahiensis (Ranzani, 1842). SUMMARY LOCALITY: Curacao, Netherlands Antilles. TYPE SPECIMEN: Holotype No. 61346, Helm. A species of Ptychobothrium was recovered Coll., USNM. from the intestine of the houndfish, Strongy-

Figs. 1-7. Plicatobothrium cypseluri. 1. Scolex in lateral aspect. 2. Cross-section of scolex near tip. 3. Immature portion of strobila showing external segmentation at intervals and early appearance of uterine sacs. 4. Proglottid near end of strobila. 5. Cross-section of mature proglottid at level of genital pore and a short distance anterior to uterine pore. 6. Reproductive complex in proglottid near end of strobila. 7. Eggs. Abbreviations: CS. cirrus sac; ET, excretory tubules; GP, genital pore; LM, longitudinal muscles; OC, oocapt; OD, oviduct; OO, ootype; OV, ovary; SR, seminal receptacle; TE, testes; UD, uterine duct; US, uterine sac; VA, vagina; VD, vas deferens; VI, vitellaria; VR, vitelline reservoir.

Copyright © 2011, The Helminthological Society of Washington 18 PROCEEDINGS OF THE [VOL. 34, No. 1 him timucii, in Jamaica, and Plicatobothrium uterine sac, and their discharge before em- cypseluri n. gen., n. sp. is described from the bryos develop hooks in Plicatobothrium. flying fish, Cypselurus bahiensis, in Curasao. Plicatobothrium and Ptychobothrium have sim- LITERATURE CITED ilar scoleces but are differentiated by large, WARBLE, R. A., AND J. A. McLEOD. 1952. The Y-shaped uterine sacs opening posteriorly, gen- zoology of tapeworms. Minneapolis, 780 p. ital pores posterior to the midlevel of the YAMAGUTI, S. 1959. Cestodes. Systema Hel- proglottids, collection of eggs largely in the minthum 2: 860 p. New York.

Morphological Differences Between the Cuticle of Swarming and Nonswarming Tylenchorhynchus martini

I. K. A. IBRAHIM*

The phenomenon of swarming in Tylen- by incubation in the following: absolute eth- chorhynchus martini, Fielding, 1956, was first anol, propylene oxide, a mixture of propylene reported by Hollis (1958), and several reports oxide and maraglas mixture 1:1 v/v, and were published subsequently on its induction maraglas mixture (Polysciences, Inc., Rydal, and nature (Hollis, 1960, 1962; Hollis and Pennsylvania). The maraglas mixture used for McBride, 1962; McBride, 1964; McBride and embedding consisted of ml proportions of Hollis, 1966). Results of the present study maraglas 34, cardolite 10, dibutyl phthalate 5, indicate that there are morphological differ- and benzyl dimethylamine 1 (Freeman and ences between the cuticle of swarming and Spurlock, 1962). A drop of fresh maraglas nonswarming populations of T. martini. Such mixture was added to the bottom of a Beem differences appear in swarming specimens as plastic capsule, then nematode specimens were morphological changes in some cuticular layers, introduced and oriented, and maraglas mixture especially the cortex and the fiber layers. These added to fill the capsule. The capsules then morphological changes do not appear in the were incubated at 60 C for 48 hr. Blocks cuticle of nonswarming specimens of T. mar- were trimmed and sections were cut to give tini. The results also indicate that the cuticle bright gold to silver interference color with a of T. martini is a multilayered structure similar glass knife in a Sorvall MT-2 Porter-Blum to that of Meloidogijne javanica (Bird and Ultra-Microtome. Sections were stained in \% Rogers, 1965) and Xiphinema index (Wright, aqueous lead oxide for 5 min and then viewed 1965). under a HU-11A Hitachi electron microscope at 50 kv. MATERIALS AND METHODS Swarming and nonswarming specimens of RESULTS T. martini, grown on rice plants in the green- Seven cuticular layers were seen, under the house, were extracted from the soil and placed electron microscope, in cross and longitudinal in 1% unbuffered osmium tetroxide solution sections of T. martini (Ibrahim, 1965). Non- for 2 hr at 5 C. Specimens were then washed swarming specimens of T. martini showed no in distilled water and dehydrated gradually morphological changes in the cuticular layers in a series of ethanol-water solutions, followed and the cuticle appeared intact (Fig. 1A). In many sections of swarming specimens morpho- * Department of Plant Pathology, Louisiana State Uni- versity, Baton Rouge. Home address: Department of Plant logical changes in the outer layers of the cuticle Pathology, College of Agriculture, University of Alexandria, UAR. were detected. These changes probably started

B FIG. 1. Tylenchorhynchiis martini, adult female. A. Longitudinal section through the cuticle of non- swarmer. B. Oblique longitudinal section through the cuticle of a swarmer, early state. Note the swelling of the outer layer at the edges of the interstrial regions (arrows). C. Oblique longitudinal section in the cuticle of swarmer, later state. D. Cross section of the cuticle of swarmer showing bright light- colored spots in the matrix and fiber layers ( arrows ) , later state. Abbreviations for figures: 1, external cortex; 2, internal cortex; 3, first boundary layer; 4, matrix; 5, second boundary layer; 6, fiber layer; 7, third boundary layer; h, hypodermis.

at the edges of the interstrial regions (Fig. IB) they reach the outer layers. Special cuticular as a swelling in the external cortex and sep- canals (Fig. 2) in the cuticle may be function- aration of the two cortical layers from each ing as avenues for the internal secretions. other. In later stages these changes extended throughout the interstrial regions (Fig. 1C). SUMMARY In some specimens, bright, light-colored spots occurred in the matrix and fiber layers in addi- Morphological differences between the cu- tion to the changes in the cortical layers (Fig. ticle of swarming and nonswarming specimens ID). of Tylenchorhynchus martini appear as mor- The morphological differences between phological changes in some cuticular layers of swarming and nonswarming specimens of T. swarming specimens. Such changes did not martini indicate that swarming may be ini- appear in the cuticle of nonswarming speci- tiated by an internal mechanism similar to mens. Morphological changes include swelling that described for hatching and molting by of the external cortex, separation of the two Rogers (1960). Internal secretions may induce cortical layers, and bright, light-colored spots the morphological changes in the cuticle as in the matrix and the fiber layers.

HOLLIS, J. P. 1958. Induced swarming of a nematode as a means of isolation. Nature 182: 956-957. . 1960. Mechanism of swarming in Ty- lenchorhynchus species (Nematoda, Tylen- chida). (Abs.) Phytopathology 50: 639. . 1962. Nature of swarming in nematocles. Nature 193: 798-799. -, AND J. M. McBRiDE. 1962. Induction of swarming in Tylenchorhynchus martini ( Nematoda, Tylenchida). (Abs.) Phytopath- ology 52: 14. IBRAHIM, I. K. A. 1965. Structure of the cuticle of Tylenchorhynchus martini (Nematoda, Ty- lenchida). (Abs.) Phytopathology 55: 1062- 1063. McBRiDE, J. M. 1964. Studies on the induction of swarming in Tylenchorhynchus martini, Fielding 1956 (Nematoda, Tylenchida). Ph.D. FIG. 2. Tylenchorhynchus martini. Oblique dissertation, Louisiana State Univ., Baton longitudinal section in the cuticle of swarming Rouge. adult female showing a cuticular canal (arrow). , AND J. P. HOLLIS. 1966. The phenomenon of swarming in nematodes. Nature 211: 545_546. LITERATURE CITED ROGERS, W. P. 1960. The physiology of infective processes of nematode parasites; the stim- BIRD, ALAN F., AND G. E. ROGERS. 1965. Ultra- ulus from the animal host. Proc. Roy. Soc. structure of the cuticle and its formation in (London), Ser. B., 152: 367-386. Meloidogt/ne javanica. Nematologica 11: 224- WRIGHT, K. A. 1965. The histology of the 230. oesophageal region of Xiphinema index FREEMAN, J., AND B. SPURLOCK. 1962. A new Thome and Allen, 1950, as seen with the epoxy embedment for electron microscopy. J. electron microscope. Canad. ]. Zool. 43: Cell Biol. 13: 437-443. 689-716.

Notes on the Genus Isolaimium Cobb, 1920 (Nematoda), with Descriptions of New Species from South Africa

WlLMA L. HOGEWIND AND J. HEYNS "

The genus Isolaimium was described by In our opinion the species described by Cobb in 1920, based on a single species I. Andrassy is not conspecific with papillatum, papillatum, collected on Plummer's Island, and is accordingly renamed Isolaimium andras- Potomac River, Virginia, USA. This species siji nom. nov., synonym Isolaimium papillatum has apparently never again been collected, of Andrassy 1962 (nee Isolaimium papillatum and no other species were added until Timm Cobb, 1920). I. andrassiji differs from I. described I. stictochroum in 1961. Shortly papillatum in the much less well-developed afterwards Andrassy (1962) gave a redescrip- transverse striae on the front part of the body. tion of /. papillatum, based on a female from Although Cobb does not mention the striae in the banks of the Adige River in Italy. his description, his drawing clearly shows very prominent striae just behind the lip region. * Plant Protection Research Institute, Pretoria, South Africa. Calculated from Cobb's drawing, these striae

Copyright © 2011, The Helminthological Society of Washington JANUARY, 1967] HELMINTHOLOGICAL SOCIETY 21 are approximately 1.2 ^ apart, while Andrassy a = 70 (49-91); b = 18 (14-23); c = 81 states that the very fine striae on his specimen (56-106); V = n-5-14-7 50 (48-53) 12.3-15.1. are only 0.8 /JL apart. Furthermore, Andrassy MALE (n = 6): L = 4.7 (3.6-5.9) mm; observed longitudinal lines on the posterior a = 75 (58-93); b = 17 (14-21); c = 75 (59- part of the body, which are apparently not 90). present in papillatum. We do not consider HOLOTYPE (female): L = 5.3 mm; a = 86; it advisable to base a redescription of papil- b = 23; c = 106; V = n-5 48 12-s. latum on specimens other than Cobb's types ALLOTYPE (male): L = 5.2 mm; a = 85; or else on topotypes, since the original descrip- b= 18; c = 78. tion is so incomplete that it is virtually im- Body lying in the shape of a letter (C) when possible to identify this species with certainty. relaxed. Body of uniform width over most of Attempts to locate the type specimens among its length, tapering only towards the front part Cobb's slides at Beltsville have thus far been of the neck and the tail. Surface of cuticle unsuccessful. marked by about 80 longitudinal striae, 1.3— Recently three species of Isolaimium were 1.8 /x apart around midbody, which occur over collected in various localities in South Africa. the entire length of the body except on the It is interesting that while papillatum and lip region. Transverse striae, also on the outer andrassyi were from aquatic surroundings, surface of the cuticle, occur from behind the stictochroum and the three South African lip region over the entire body. These striae species are from cultivated fields. are especially prominent just behind the lip The taxonomic position of Isolaimium was region, where they are about 1.6—2 ^ apart. discussed by Timm (1961) and Andrassy Towards the base of the esophagus they be- (1962). There is nothing we can add to this came closer together and less prominent, and except to endorse Andrassy's view that this even more so over the rest of the body. On genus probably deserves to be put in a family the front part of the neck the cuticle is divided of its own. The peculiar nature of the cephalic into distinct blocks by the longitudinal and papillae has already been remarked on by transverse striae. Head only slightly offset by Andrassy. Face views prepared of two of a shallow depression. Lips not distinct rather the South African species shed further light closely amalgamated, the contour somewhat on the nature of these papillae. Normally one hexagonal in a face view. Lip region 21—27 /j, finds nerve fibers leading inwards from ce- wide, about half as wide as the body at the phalic papillae, but in this case there appears base of the esophagus. In the subcuticle just to be thick-walled tubes leading backwards behind the lip region there occurs a darkish from the inner circle of six "papillae" (see colored pocl-shaped organ on either side of the Fig. IB, C, E). In a face view a hollow neck. Stoma 95 to 134 ^ long, cylindrical, lumen can be seen in these tubes. Two of with strongly cuticularized walls. In cross these tubes come very close to the sensillae section the stoma consists of three cuticularized pouches, but no definite connection with the elements, forming a roughly triangular lumen. latter could be traced. Cobb could not locate Stoma 3.6-4.9 times as long as width of lip the amphid apertures in his species, and neither region. Esophagus cylindroid, about one-third could Andrassy find the amphicl apertures on the corresponding body diameter, and 145- his specimen. In the three species which we 190 p. in length. Cardia elongate-conoid. studied the amphid apertures could also not Nerve ring situated at about 44% of length of be found. On the other hand, Timm ob- esophagus, measured from base of stoma. Tail served small pore-like amphid apertures on dorsally convex, ventrally concave, bluntly I. stictochroum. rounded, 48-69 ^ long, 1.0—1.4 times the anal body diameter. Subcuticle thickened around DESCRIPTION OF NEW SPECIES terminus of tail, and longitudinal striae forming very distinct annules around the tail tip. Isolaimium africanum n. sp. No caudal papillae observed in the female, (Fig. 1A-H, Fig. 2B, C) male with one postanal ventromedian papilla FEMALE (n = 6): L = 4.8 (3.9-5.7) mm; and a pair of subclorsal papillae. Rectum

A-D

FIG. 1. Is'olaimium africanum n. sp. A-D. Face view at level of cephalic papillae, slightly further backwards, middle of stoma, and esophagus, respectively. E. Head. F. Posterior part of male. G. An- terior part of body. H. Vulva and vagina.

FIG. 2. A, D-F. Isolaimhim multistriatum n. sp. A. Anterior part of body. D. Vulva and vagina. E. Female tail. F. Head. B, C. Isolaimium africanum n. sp. B. Male tail. C. Female tail.

FIG. 3. Isolaimium incus n. sp. A-C. Face view at level of eephalic papillae, slightly farther backwards, and at middle of stoma, respectively. D. Anterior part of body. E. Head. F. Male tail. G. Posterior part of male.

Copyright © 2011, The Helminthological Society of Washington JANUARY, 1967] HELMINTHOLOGICAL SOCIETY slightly longer than anal body diameter. Pre- a = 71-74;b=18 (14-19); c = 84 (61-93); rectum not observed. No lateral field. Width V = 50 (49-53). of lateral cord equal to one-third the body HOLOTYPE (female): L = 5.0 mm; a = 74; diameter. b = 19; c = 92; V = 14-5 53 15-8. Female amphidelphic. Vulva large, circular, Description similar to that of Isolaimium sunken below the surface, with small inner lips; africanum except for the following differences: vagina short, broad; gonads clorylaimoid in Lip region not offset, but shaped as in /. appearance, with large uiidifferentiated uterus, papillatum, with the transverse striae begin- separated from oviduct by sphincter muscle; ning somewhat further backwards than in ovary reflexed. Uterine egg measures 65—70 by 38-42 p., with egg shell 3 ^ thick. africanum. Transverse striae much less dis- Spicules strongly arcuate, measuring 52-61 tinct. Longitudinal striae at least 120 in num- H along the curved median line. No lateral ber and about 1 p. apart. Stoma 100-106 p. guiding pieces. Gubernaculum thin, linear, long. Tail convex—conoid, more pointed than more than one-third as long as spicules, and in africanum. Caudal papillae observed near provided with a prominent backward-directed the tail terminus. Several eggs occur together process at its proximal end. No adanal sup- in the uterus, eggs measuring 78-84 by 42 p.. plements, but a ventromedian series of two Male not known. to six, irregularly spaced. TYPE LOCALITY AND HABITAT: Four females TYPE LOCALITY AND HABITAT: The type and several juveniles were collected in a maize specimens were collected during September field on the farm "Rustig" of Mr. C. Kriel in 1963 and again in June 1964 on various local- the Clocolan district of the Orange Free State, ities on the farm Elandsfontein of the South December 1963. African Forest Investments Co., near Sabie in TYPE SPECIMENS: Holotype on slide 4010, the Transvaal, and were found in soil in Finns paratypes on slide 4017. patula and Eucalyptus saligna plantations. DIAGNOSIS: I. multistriatum n. sp. differs TYPE SPECIMENS: Holotype on slide 4667, from its nearest relative I. africanum n. sp. in allotype on slide 2897, paratypes on slides the characters mentioned above. 2896, 2913-2916, 4666, 4668, and 4670. One female paratype deposited in the collection Isolaimium incus n. sp. of the USDA, Nematology Investigations, (Fig.3A-G) Beltsville, Maryland. DIAGNOSIS: I. africanum n. sp. differs from MALE (n = 3): L = 3.0-3.7 mm; a = 79- I. papillatum Cobb and I. andrassiji nom. nov. 87; b = 16; c = 71-79. in the presence of longitudinal striae on the HOLOTYPE (male): L — 3.7 mm; a = 87; surface of the cuticle. From /. andrassyi it b =16; c = 79. further differs in the presence of prominent Body ventrally curved, especially in pos- transverse striae on the front part of the neck, terior part; of uniform width over most of and in the shape of the tail. From I. sticto- its length, tapering only towards front part chroum it differs in the shape of the lip region of neck and tail. Surface of cuticle with about which is confluent in stictochroum but slightly 50 longitudinal striae, 1.3—1.5 p, apart around offset in africanum, and in the presence of midbody, occurring over the entire length of prominent transverse striae on the surface of the body except on the lip region. Surface the cuticle. From multistriatum n. sp. it of cuticle also marked by transverse striae, but differs in the smaller number of longitudinal these are very indistinct, even on the front striae, differently shaped tail, and slightly part of the neck. Lip region truncate, ex- offset lip region. panded, set off by broad depression. Lips rather well separated, hexagonal in a face Isolaimium multistriatum n. sp. view. Lip region 22 p, wide, which is two- (Fig. 2A, D-F) thirds the width of the body at the base of FEMALE (n = 4): L = 4.8 (4.1-5.2) mm; the esophagus. Stoma 84 p, long. Esophagus

Copyright © 2011, The Helminthological Society of Washington 26 PROCEEDINGS OF THE [VOL. 34, No. 1 cylindroid and about 135 /jt long. Cardia heart- KEY TO SPECIES OF Isolaimium shaped, about 12 ^ long. Nerve ring situated 1. Longitudinal striae absent, or present at 47% of the length of the esophagus. Tail only on posterior part of body 2 conoid, dorsally convex, ventrally concave, the Longitudinal striae over entire body ____ 3 terminus bluntly rounded. Tail 37-47 ^ long, 2. Transverse striae conspicuous behind 1.2-1.4 times the anal body diameter. Sub- lip region papttlatum cuticle slightly thickened around terminus of Transverse striae very fine andrassyi tail. Tail with a postanal ventromedian papilla 3. Lip region expanded and set off by a and two subterminal pairs of papillae. No pronounced depression incus lateral field. Lateral cord about one-fourth as Lip region not expanded, confluent, or wide as the body. set off by a slight depression only ____ 4 Spicules strongly arcuate, measuring 44-47 4. Fine transverse striae in subcuticle ____ ju, along the curved median line. No lateral stictochroum guiding pieces. Gubernaculum more than one- Conspicuous transverse striae on surface third as long as spicules, thin, linear, and proof cuticle 5 vided with a small backward-directed process 5. Longitudinal striae more than 120; tail at its proximal end. No adanal supplements, conoid multistriatum but a ventromedian series of three to five, the Longitudinal striae about 80; tail bluntly first of which occurs opposite the middle of rounded africanum the spicules. Female not known. LITERATURE CITED TYPE LOCALITY AND HABITAT: Collected in ANDRASSY, I. 1962. Nematoden aus dem Psam- a maize field in the Bothaville district in the mon des Adige-Flusses II. Mem. Mus. Civico Orange Free State, December 1964. St. Nat. Verona 10: 1-35. TYPE SPECIMENS: Holotype on slide 5535, COBB, N. A. 1920. One hundred new nemas. paratypes on slides 5536 and 5682. Contr. Sci. Nematol., IX. Baltimore, p. 217- DIAGNOSIS: Isolaimium incus n. sp. can be 343. TIMM, R. W. 1961. The systematic position of distinguished by its expanded lip region, which Isolaimium Cobb, 1920 (Nematoda), with a is different from any other species in the description of a new species. Jl. Bombay genus. Nat. Hist. Soc. 58: 302-304.

Experimental Transmission of Histotnonas meleagridis and Heterakis gallinarum by the Sow-bug, Porcellio scalier, and Its Implications for Further Research

L. A. SPINDLER*

The epizootiology of poultry blackhead has that H. meleagridis (histomonads) can be not been completed, especially as it concerns transmitted from bird to bird by the embryo- the modes of natural transmission of the causal nated eggs or larvae of a parasitic invertebrate, agent, the protozoon, Histomonas meleagridis. the poultry cecal nematode, Heterakis galli- Studies by Graybill and Smith (1920), Smith narum (heterakids). Other investigations dis- and Graybill (1920), and others established closed that heterakids and histomonads, and * Beltsville Parasitological Laboratory, Animal Disease therefore blackhead, can be disseminated by and Parasite Research Division, Agricultural Research Ser- vice, U.S. Department of Agriculture, Beltsville, Maryland. certain free-living invertebrates. For example,

Copyright © 2011, The Helminthological Society of Washington JANUARY, 1967] HELMINTHOLOGICAL SOCIETY 27 infections of one or the other or both parasites, smears in physiologic saline were prepared with or without signs of blackhead, were ob- from the contents of each cecum and examined served in birds after they were fed (a) earth- for histomonads and other protozoa by phase- worms and houseflies from poultry or pheasant contrast microscopy. The remainder of the yards (Curtice, 1907; Ackert, 1917; DeVolt contents and the scraped-off mucosa were sus- and Davis, 1936; Madsen, 1962, Lund, Wehr, pended together in warm saline, sedimented, and Ellis, 1963), and (b) houseflies, fleshflies, and examined for heterakids; appropriate ex- and grasshoppers that had been kept in contact aminations were made for other helminths. with embryonated eggs of heterakids obtained EXPOSURE OF SOW-BUGS TO PARASITES: Each from sources where blackhead was present day for 2 weeks, 1 ml of a rich suspension of (Frank, 1953). heterakid eggs in water was distributed on This information makes suspect as a poten- the meat, on the underside of the wood, and tial disseminator of these parasites any free- on the soil of each terrarium. These eggs were living invertebrate that may be eaten by poul- from pooled cultures that were being used try. One such invertebrate is Parcellio scaber, successfully to transmit heterakids and histo- the sow-bug (bugs), a crustacean that feeds monads to poults and which had produced on decaying or putrefying organic matter, and clinical blackhead in them under experimental is commonly found in poultry yards. Such conditions. bugs might serve as sources of infection for TESTS OF TRANSMISSION: After exposure, 16 birds that swallow them. Since this possibility bugs of Lot I and 145 of Lot II were washed has not been tested, a study was made to de- in wire baskets for 5 min under a strong stream termine if P. scaber could transmit heterakids of warm water from a faucet, then in five and histomonads to turkeys under experimental changes of water in beakers to dislodge eggs conditions. that might be adhering to the bugs. Those of Lot I were then force-fed, alive, to 8 poults, MATERIALS AND METHODS 2 per bird; those of Lot II were force-fed, alive, TURKEYS: Beltsville Small White poults, 6 to 29 poults, 5 per bird. Fifteen other poults weeks old, were used. From hatching to ne- not fed bugs were kept as controls. The cropsy they were maintained in wire-bottom test and control birds were observed for signs cages, under conditions designed to preclude of blackhead for 14 days and were then extraneous infections of histomonads and necropsied. heterakids. Clean feed and water were pro- MICROSCOPIC EXAMINATION OF SOW-BUGS vided throughout. AND TESTS ON TERRARiA: After the bugs ex- SOW-BUGS: Two lots (I and II) of adult posed to parasites were fed to poults, five sow-bugs were collected, each from a single remained in each terrarium. They were trans- "wild" colony. Each lot of bugs was main- ferred to clean terraria for 7 days. They were tained in a separate terrarium, a 500-ml then dissected and examined microscopically beaker. The soil was overlaid with several for eggs and larvae of heterakids and for histo- small pieces of partially decayed wood. To monads to indicate the mode of transmission. maintain moisture, the underside of the wood The surface soil and scrapings from the and the soil were sprayed lightly with water underside of the wood of each original terrar- as needed. Scraps of putrefying raw beef for ium were mixed in feed and administered to food were placed each day beneath wood a poult that was later examined post-mortem. where the bugs congregated. This examination indicated whether the para- TESTS FOR NATURAL INFECTIONS OF PARA- sites were viable during the time the bugs SITES: Thirty-four bugs of Lot I were force- were associated with them. fed, alive, to 17 poults, 2 per bird; 50 of Lot II were force-fed, alive, to 10 poults, 5 per RESULTS bird. Sixteen days later, necropsies were per- All the birds fed sow-bugs and the ones formed on the birds. not fed these crustaceans remained healthy and NECROPSIES: Livers and ceca were inspected signs of blackhead were not detected. All for signs of blackhead. For each bird, two transmissions that were obtained were of histo-

Copyright © 2011, The Helminthological Society of Washington 28 PROCEEDINGS OF THE [VOL. 34, No. 1 monads and heterakids together. The amount of 37 poults fed bugs exposed to these para- of transmission was small, about the same for sites became infected. The infections were the two lots of bugs. For that reason the find- small, and none of the birds developed signs ings are treated as a unit; the pertinent ones of blackhead. The bugs had been given a are listed below. good opportunity to ingest sizable numbers of heterakid eggs, some of which must have con- Birds fed sow-bugs exposed to tained histomonads. This is indicated by the heterakids and histomonads fact that poults fed materials from the terraria Number of birds 37 used to expose the bugs developed blackhead Number parasite-free at necropsy 29 and died, and large infections of heterakids Number with blackhead signs 0 and histomonads were found in them. Number with heterakids and histomonads The failure to find heterakids and histom- at necropsy 8 onads in poults fed sow-bugs not exposed to Histomonads in cecal smears 1/8—10 fields these parasites indicates that natural infections Heterakids per bird 1-3 of them did not exist in the colonies from Birds with heterakids of both sexes 4 which the bugs were collected. Such transmission as was obtained prob- Birds fed sow-bugs not exposed to heterakids and histomonads ably was from heterakid eggs in the gut of the bugs at the time they were fed to poults, and Number of birds 27 was therefore mechanical; there was no indica- Number with parasites 0 tion that the bugs served as a host for the Number with blackhead signs 0 heterakids and histomonads. Neither nema- Controls (birds not fed sow-bugs) tode eggs nor larvae nor histomonads were found in the 10 exposed bugs that were dis- Number of birds 15 Number with parasites 0 sected after they had been kept for 7 days Number with blackhead signs 0 in clean terraria. Had the eggs hatched in the gut of the bugs and the larvae migrated No helminth parasites other than heterakids to the surrounding tissues, they should have were found; protozoa other than histomonads, persisted there and some should have been if present, did not occur in numbers sufficient found. P. scaber is capable of serving as an to be revealed by the method of examination intermediate host as was shown by Cram employed. (1930) in the case of Disphanjnx spiralis, a The two birds fed materials from the orig- nematode parasite of birds. inal terraria developed blackhead and died. In these attempts at transmission, there At necropsy, signs of blackhead were observed were factors that may have influenced the in each, and the cecal contents contained mas- results. Small, presumably young, bugs be- sive numbers of histomonads; 193 heterakids came numerous in the terraria, but in spite were recovered from one of the birds, 247 of this increase the number of bugs in each from the other. terrarium decreased with time. When trans- Neither nematode eggs or larvae nor histom- mission feedings were made, it was necessary onads were found by dissection and micro- to use some smaller bugs as well as some larger ones. It is not known if the smaller bugs scopic examination of the five bugs of each swallowed eggs in any numbers. Another po- collection that were kept in clean terraria for tential factor is the proportion of the heterakid 7 days after exposure to eggs of heterakids. eggs that may be expected to have contained DISCUSSION histomonads, about one in 200 according to Lund and Burtner (1957). This may have Under conditions of this study, the sow-bug, influenced transmission, especially if small Porcellio scaber, did not prove to be a good numbers of eggs were contained in the gut transmitter of the nematode, Heterakis gal- of individual bugs at any one time. linarum, and the protozoon, Histomonas melea- Although the amount of transmission that gridis, and therefore blackhead. Only eight was obtained was too small to appear impor-

Copyright © 2011, The Helminthological Society of Washington JANUARY, 1967] HELMINTHOLOGICAL SOCIETY 29 tant, there may be a long-range significance. 15 others not fed bugs (the controls) remained Of the eight birds that became infected, four healthy and free of parasites. Neither nematode harbored heterakids of both sexes. If these eggs nor larvae nor histomonads were found worms had been permitted to develop to sex- in 10 exposed bugs that were kept in clean ual maturity, fertile eggs, some with histom- terraria for 7 days, then dissected for micro- onads, might have been produced. Therefore, scopic examination. Apparently, the transmis- from a small beginning, such as produced by sion obtained was mechanical, from unhatched one or a few sow-bugs, the infections might, heterakid eggs in the gut of the bugs at the in time, build up to serious proportions under time they were fed to poults. Some implica- flock conditions. tions for further research on transmission are There are other potential avenues of trans- discussed. mission of heterakids and histomonads, and therefore blackhead, that may offer implica- LITERATURE CITED tions for further research. These include those ACKERT, J. E. 1917. A means of transmitting free-living invertebrates and those ectoparasites the fowl nematode, Heterakis papillosa Bloch. that at some stage of their development may Sci. N. S. 46 (1190): 394. be coprophagous. Some histomonads in heter- CRAM, E. B. 1930. Porcellio scaber as the inter- akids are said to occur in the reproductive mediate host of Dispharynx spirali-s of birds organs, and may have an affinity for repro- in this country. J. Parasit. 16(3): 169. ductive tissues. If that is the case, the proto- CURTICE, D. 1907. Notes on experiments with zoon may have an affinity for the reproductive blackhead of turkeys. U.S. Dept. Agric. Bur. An. Ind. Circ. 119: 1-10. tissues of other gastrointestinal helminths as DEVOLT, H. M., AND C. R. DAVIS. 1936. Black- well. Until all forms capable of carrying his- head (infectious enterohepatitis) in turkeys, tomonads and heterakids are identified, and with notes on other intestinal protozoa. The their potential for transmission evaluated, it Univ. Maryland Agr. Exp. Sta. Bull. (392): may be impossible to develop measures of 493-567. maximum efficiency for the control and elim- FRANK, J. F. 1953. A note on the experimental ination of blackhead. transmission of enterohepatitis of turkeys by arthropods. Canad. J. Comp. Med. and Vet. SUMMARY Sci. 17 (5): 230-231. GRAYRILL, H. W., AND T. SMITH. 1920. Pro- Sow-bugs, Porcellio scaber, were tested ex- duction of fatal blackhead in turkeys by feed- perimentally for their ability to transmit the ing embryonated eggs of Heterakis papillosa. protozoon, Histomonas meleagridis, and its J. Exptl. Med. 31(5): 647-655. vector, the parasitic nematode, Heterakis galli- LUND, E. E., AND R. H. BURTNER, JR. 1957. In- narum. Sow-bugs from natural colonies were fectivity of Heterakis gallinae eggs with His- tomonas meleagridis. Exptl. Parasit. 6(2): housed in terraria, exposed to histomonad- 189-193. bearing eggs of heterakids, and fed to suscep- , E. E. WEHR, AND D. ELLIS. 1963. Role tible poults. None of the 37 birds developed of earthworms in transmission of Heterakis signs of blackhead; at necropsy, 8 harbored and Histomonas to turkeys and chickens. ]. light infections of heterakids (1 to 3 per bird) Parasit. 49(5, Section 2): 50. and histomonads (1 per 8 to 10 microscope MADSEN, H. 1962. On the interaction between fields in cecal smears). Two other poults fed Heterakis gallinarum, Ascaridia galli, "Black- materials from the experimental terraria died head" and the chicken. J. Helminthol. 36 (1/2): 107-142. of blackhead, and harbored massive infections SMITH, T., AND H. W. GRAYBILL. 1920. Black- of histomonads and heterakids, showing that head in chickens and its experimental produc- the parasites used were viable. Twenty-seven tion by feeding embryonated eggs of Heterakis birds fed bugs not exposed to parasites, and papillosa. J. Exptl. Med. 32(2): 143-152.

Morphological Variant of Tetylenchus joctus Thorne (Nernata: Tylenchida) Associated with Cultivated Blueherries in Indiana"

VIRGINIA R. FERRIS AND JOHN M. FERRIS!

Tetylenchus joctus Thome, 1949, has been peat soil, and both from around plants of the reported to be frequently associated with the variety Jersey. roots of cultivated blueberries (V actinium One-pint subsamples of each soil sample conjmbosum L.) in New Jersey (Hutchinson were processed using a combination of sieves et al, 1960; Hutchinson et al, 1961), and and Baermann funnels. Aliquots of the nem- with cranberries (V. macrocarpon Aiton) in atodes recovered were counted, and the counts Massachusetts (Zuckerman 1960, 1961). Hutch- averaged. Nematodes which were to be mea- inson and his co-workers (1961) reported find- sured were relaxed by gentle heat, killed, and ing very high populations (up to 6,000/pint fixed in formalin-acetic-alcohol (FAA), de- of soil) in one planting of blueberries near hydrated to pure glycerine, and mounted in New Lisbon, New Jersey, where the popula- glycerine using Thome's (1961) methods. tion remained abundant over a 4-year period. Zuckerman (1961) has shown that T. joctus RESULTS feeds on the epidermal cells of the roots of Very few plant parasitic species were re- cranberry plants, causing a collapse of the covered from the soil samples, with the excep- cells. Though the cultivation of blueberries tion of the sample from North Judson which constitutes an important industry in Michigan, was taken from the 10-year-old planting of the and a growing industry in Indiana, this species variety Jersey in the dark sandy soil. In this has not previously been reported to be asso- soil sample there were present an average of ciated with the crop in either of these states. 7,000 specimens of a Tetylenchus species per Fairly extensive sampling of soil in blueberry pint of soil. No specimens of Tetylenchus plantings has been carried out in Michigan were recovered from any of the other soil (Knierim, 1960). samples.

MATERIALS AND METHODS IDENTITY OF THE Tetylenchus SPECIES Soil samples were taken in July and August Examination of the specimens showed that 1965 from blueberry plantings in two locations this was a species similar to T. joctus; however, in northern Indiana. On one farm in North certain details of the tail in both males and Judson, Indiana, the following soil samples females did not follow the description. T. were taken, by means of a sampling probe, joctus was described as having the terminus from around the base of plants in different subacute, and with no striations on the term- areas of the farm: (1) dark sandy soil, blue- inus (Thorne, 1949). The shape of the tail berry variety Jersey, planted about 10 years was given as one of the diagnostic characters previously; (2) dark sandy soil, blueberry vari- of the species. The specimens from Indiana, ety Burlington, planted about 17 years pre- however, have a terminus that is usually acute, viously; (3) black sandy soil containing more sometimes almost mucronate, as was described organic matter (muck) than the first two for T. productus Thorne, 1949. In addition, samples, blueberry variety Jersey; and, (4) in the Indiana specimens the termini are uncultivated dark sandy soil, wild blueberries. striated and are similar in appearance to those At the other location (La Paz, Indiana) two described for T. annulatus Merny, 1964. The soil samples were taken, both a highly organic tail shape is similar for both males and females (Fig. 1). ::: Journal Paper No. 2700, Purdue University, Agricul- tural Experiment Station. In other characteristics the Indiana speci- f Department of Entomology, Purdue University, Lafay- ette, Indiana. mens follow closely the description for T.

FIG. 1. Variations in tail observed in Indiana specimens of T. joctus. A-F, female tails; G—I, male tails.

Copyright © 2011, The Helminthological Society of Washington 32 PROCEEDINGS OF THE [VOL. 34, No. 1 joctus, a species originally described from tail features described above. However, we specimens collected in Wrangell, Alaska. A have ascertained by correspondence with Pro- comparison of the standard measurements and fessor Gerald Thome, who has examined speci- ratios is given: mens from a New Jersey population, and from Indiana, that all of these specimens from both Tetylenchus joctus Thorne, 1949 localities exhibit the atypical tail characteristics FEMALE: 0.7 mm; a = 30; b = 4.5; c = 9.4; which we have described here on the basis of Indiana materials. This strengthens the belief V = »i5538; stylet: 15 p.. MALE: 0.6 mm; a = 30; b = 4.5; c = 9.0; that the species has been introduced to Indiana. T = 70. SUMMARY Tetylenchus joctus, Indiana Specimens Although Tetylenchus joctus Thorne, 1949, FEMALE (n =10): 0.67mm (0.61-0.73); has been reported to be a frequent associate a = 31 (28-34); b = 5.4 (4.6-6.1); c = 12 of blueberries and cranberries in the eastern United States, it has not previously been dis- (10-18); V = 2(i56--1 (23-3255_5Q21-27). stylet = 15 ^ (15-16). covered in blueberry plantings in the north MALE (n=10): 0.60 mm (0.56-0.64); central region. Specimens recovered in large a = 31 (28-39); b = 4.8 (4.5-5,3); c = 10 numbers from soil around blueberry plants in (9-12); T = 45 (43-50); stylet = 15 p (15- one location in Indiana differed in certain characteristics from T. joctus as originally de- 16). Because of the similarity of the Indiana scribed. These differences are detailed and specimens to T. joctus, it has been decided to the population recorded as a morphological consider this geographically disjunct popula- variant of T. joctus. tion as a morphological variant of that species and to record herewith the variations described LITERATURE CITED above. HUTCHINSON, M. T., J. P. REED, AND S. R. RACE. 1960. Nematodes stunt blueberry plants. New DISCUSSION Jersey Agric. 42(4): 12-13. , , H. T. STREU, A. A. DiEmvARDO, The fact that T. joctus has not previously AND P. H. SCHROEDER. 1961. Plant para- been reported in the north-central part of the sitic nematodes of New Jersey. New Jersey U.S., though it appears to be a frequently Agric. Exp. Sta. Bull. 796: 1-33. encountered species in blueberry and cran- KNIERIM, J. A. 1963. Nematodes associated with berry plantings in the East (Mai et al, 1960), crop plants in Michigan. Michigan Agric. leads us to suspect that the species was im- Exp. Sta. Quart. Bull. 46(2): 254-262. MAI, W. F., H. W. CRITTENDEN, AND W. R. ported to Indiana on blueberry plants. The JENKINS. 1960. Distribution of stylet-bear- Indiana grower who owned the planting in ing nematodes in the northeastern United which the species was found reported that the States. New Jersey Agric. Exp. Sta. Bull. particular area in question had been planted 795: 1-65. before he acquired the property, but that he MERNY, G. 1964. Un nouveau Tylenchida was certain that all of the original seedlings d'Afrique Tropicale: Tetylenchus annulatufi had come from either Michigan or New Jersey. n. sp. Nematologica 10: 425-430. Our hypothesis tends to be confirmed by the THORNE, G. 1949. On the classification of the Tylenchida, new order (Nematoda, Phas- fact that we have not found the species midia). Proc. Helminthol. Soc. Washington around wild blueberries, even in close proxim- 16: 37-73. ity to the infested cultivated area. T. joctus . 1961. Principles of nematology. Mc- has been found in association with wild blue- Graw-Hill Book Co. Inc., New York, 553 p. berries in New Jersey (Hutchinson et al., ZUCKERMAN, B. M. I960. Parasitism of cranberry roots by Tetylenchus joctitfi Thorne. 1961). Nematologica 5: 253-254. There is no reference in any of the published . 1961. Parasitism and pathogenesis of records of the occurrence of this species in the cultivated cranberry by some nematodes. New Jersey or Massachusetts to the atypical Nematologica 6: 135-143.

Notes on the Life History of Pleurogonius malaclemys Hunter, 1961 (Trematoda: Pronocephalidae) from Beaufort, North Carolina, with a Description of the Cercaria

WANDA SANBORN HUNTER*

after removal from their cysts. These were photographed alive and then preserved in hot A monostome cercaria developing in the AFA solution under minimum cover glass pres- marine snail, Nassarius obsoleta (Say), in the sure. Adult worms from experimentally fed Beaufort, North Carolina area, is proved to turtles also were photographed before killing be the larva of Pleurogonius malaclemys in hot AFA. Measurements of both the meta- Hunter, 1961. This is the first marine cercaria cercariae and adults were made on stained definitely determined to be the larva of an and mounted specimens. Whole mounts were adult of the Family Pronocephalidae Looss, stained with either Semichon's or Ballard's. 1902. Morphology of the metacercariae which Photomicrographs of all living stages were commonly are encysted on and under the made with a Polaroid camera; this method opercula of the snail host, as well as results is particularly good for recording details of of feeding experiments with the definitive the excretory system which constantly vary host, Malaclemys terrapin centrata (Latrcille), in visibility. 1802, validate the above statement. DESCRIPTION OF CERCARIA MATERIALS AND METHODS Measurements in mm from 10 heat-killed Active, freshly emerged cercariae were specimens: killed in boiling seawater and measured under cover glass with the minimum amount of Range Average water necessary to prevent distortion from Body length 0.493-0.554 0.525 pressure; this method is that used by Cable Body width ( immediately (1956). Mature living cercariae could not be posterior to collar ) 0.208-0.281 0.227 studied successfully under cover slip pressure Oral sucker due to their habit of rapid encystment when Longitudinal diameter 0.040-0.054 0.049 in contact with any suitable substrate. To Transverse diameter 0.038-0.054 0.045 facilitate observation of internal organs, cer- Tail length 0.478-0.622 0.572 cariae were allowed to encyst on a slide and Tail width ( near base ) 0.053-0.068 0.059 dissected out of their cysts within 2 or 3 min. This method allowed most of the cystogenous Large, trioculate, monostome with smooth cells to empty, but did not alleviate entirely cuticle. Cephalic collar inconspicuous with the difficulties of observation due to body pig- ventral lobes approximately one-fifth to one- ments. Development of the excretory system fourth of body length; lobes widely separated was studied in living worms obtained by crush- posteriorly, but joined medially behind oral ing the snail hosts; young, developing cercariae sucker. Heavily pigmented body tends to be were dissected out of the rediae. Sections of concave ventrally. Anterior end bluntly ta- mature as well as developing cercariae were pered with terminal, weak, subspherical oral sucker; body widens abruptly posterior to made; hematoxylin and eosin staining was eyespots. Sides straight from posterior collar used. margins to rounded posterior end. Inconspic- Metacercariae of different ages were studied uous cup-shaped protrusible, apparently glandular, locomotor structures at posterolateral ::: Department of Zoology, Duke University, Durham, North Carolina; Duke University Marine Laboratory, Beau- borders of body, extending beyond level of fort, North Carolina. This study was supported by the Duke University Re- tail insertion. Cystogenous glands, filled with search Council. Mrs. Doris H. King is acknowledged for assistance with illustrations. granular opaque material, immediately be-

Copyright © 2011, The Helminthological Society of Washington JANUARY, 1967] HELMINTHOLOGICAL SOCIETY 35 neath entire body surface except for extreme bladder, or vestibule; this alternates in pulsa- anterior end. Six cephalic glands surround oral tions with thicker-walled posterior bladder. sucker and their ducts open at extreme anterior (See Fig. 7, 8, 11, 14, 15, 16.) Therefore, end (Fig. 1, 3, 29). Median eyespot smaller, bladder variable in shape, definitely of two more irregular, less organized, usually lacks parts, and when empty disappears from view lens, and lies slightly anterior to other two, and arms of large collecting ducts may then close to oral sucker. The three eyespots closely be easily mistaken for parts of a Y-shaped associated with bilobed ganglionic mass of bladder. In a mature cercaria, the heavy- nervous system (Fig. 3, 18, 29). lipped excretory pore is in the dorsal wall of Pharynx lacking, esophagus thin, long, bi- the bladder, some distance from the posterior furcating posterior to collar in second one- margin of the body (Fig. 26); it is nonfunc- fourth of body length. Cecae long, with irreg- tional until the tail is lost. Small caudal atrium ular thick-walled cliverticula throughout their present; its two posterolateral openings func- length; diverticula more prominent and reg- tion as excretory pores until tail is cast off ular in anterior parts of cecae (Fig. 5, 17, 26). at encystment (Fig. 10, 11, 26). Caudal ex- Primordia of all genital organs present; two cretory tubes prominent in developing forms small extratesticular masses widely separated (Fig. 7, 8), but at emergence, no evidence of from each other in posterior part of body (Fig. a caudal duct or pores. Branches of main col- 9, 26, 27); primordia of vitellaria anterior and lecting ducts as described for adult (Hunter, lateral to testes (Fig. 26, 27). A more con- 1961) visible in living forms. Flame cell pat- spicuous median line of cells (Fig. 5, 6, 17, tern expressed as 2(3 + 3 + 3) + (3 + 3 + 3) 18, 26) extends anteriorly from position of (Fig. 28). future adult ovary-ootype complex (Fig. 5, 6, Tail straight, slightly longer than body, 26, 27); this begins in front of excretory blad- tapering gradually to tip. Few, irregularly der and runs into anterior half of body where scattered, small cells throughout length. Few, it turns to left and ends in post-bifurcal region long, and somewhat spiral strands run length- of future genital pores. Rudiments of male wise, independent of scattered cells. Cuticle and female ducts (Fig. 2), including one inter- with fine striations. preted as Laurer's canal (Fig. 6, 26, 27) (not Development occurs in simple rediae (Fig. observed in adult, Hunter, 1961), as well as 12, 13, 14) which contain much yellow—orange location of future genital pores, can easily be and brown pigment, and which have a rela- traced in sections (Fig. 4, 26, 27). tively long, wide gut. Young rediae often show Most prominent part of excretory system marked constriction near anterior end. Birth consists of two lateral collecting tubes, filled pore inconspicuous, relatively close to pharynx. with refractile bodies. In collar region, these Cercariae leave rediae at early stage and ma- ducts unite by a very small tubule to complete ture in tissue spaces of host. Usually but two cycloid system characteristic of Family (Fig. or three (often only one) cercariae were found 26, 28). Posteriorly large ducts meet and developing in each redia; several small germ connect with bladder by thin-walled duct balls present in rediae with advanced larvae. which regularly expands to form an anterior No second-generation rediae found.

FIGS. 1-15. 1. (Frontal sec.) Ducts of cephalic glands. (200x). 2. (Cross sec.) Male and female ducts immediately posterior to developing cirrus pouch. (970x). 3. (Frontal sec.) Cephalic glands, ganglia, eyespots. (430x). 4. (Sagittal sec.) Cirrus pouch and genital pore area. (970x). 5. (Fron- tal sec.) Gut, genitalia, bladder. (200x). 6. (Sagittal sec.) Genital ducts, ootype region, Laurer's canal. (430x). 7. Live, developing cercaria ex snail. Thick-walled part of excretory bladder and caudal ducts. (150x). 8. Live, developing cercaria. Bladder and caudal ducts. Note contracted anterior bladder. (200x). 9. (Frontal sec.) Bladder, ducts, testes, locomotor organs. (430x). 10. (Fron- tal sec.) Bladder, atrium, and atrial pores. (430x). H. Live cercaria ex redia. Two parts of bladder and atrial pores. (lOOx). 12. Live redia. (48x). 13. Live redia with young cercariae. Note constriction. (48x)- 14. Live redia with older developing cercariae. Note double excretory bladder. (48x). 15. Live cercaria within redia. Note thicker wall of posterior part of bladder. (150x).

DISCUSSION OF CERCARIA Faust, 1917, are very numerous and prominent in the tail of developing forms (Fig. 19, 20, Infected snails with emerging cercariae are 21). These become smaller and less prominent to be found throughout the year, being some- in the tail of older developing and actively what more prevalent during the late fall and swimming forms. Kruidenier and Mehra (1957) early months. The percentage of infected snails described the distribution and character of the with emerging cercariae when isolated in the mucoid glands in the freshwater pronocephalid laboratory is usually less than 1%, although Macrovestibiilum eversum Hsii. They found two collections (in November and December 10 pairs of irregular glands in the tail. Kruid- of different years) from isolated pens where enier, 1953, postulates the function of the infected turtles had been confined showed body and tail gland secretions to be involved unusually high 3.3 and 4% infections. In any with the cuticle. No histochemical studies area where infected snails are found, cysts have as yet been carried out by me on Cercaria are conspicuous on and under opercula of both P. malaclemys. However, Fig. 21 does suggest infected and noninfected snails. that there is some secretion from cells within In the laboratory, midday is the optimum the tail. No constant number of these caudal time for emergence of cercariae; relatively few bodies is to be found; in young developing emerge daily as would be expected from the forms the tails are packed with them (Fig. observations of the numbers developing within 19). I suggest that certain of these bodies each redia. The cercariae are comparatively may be homologous to the glycogen-containing slow, often steady swimmers, moving in circles caudal vesicles of certain cercaria including by means of violent tail-lashings. When they Notocotylus which were described by Genet- stop swimming, they soon encyst on snails as zinskaja and Dobrovolski, 1962. They, there- well as on the bottom of the glass culture fore, logically would disappear as the actively dishes in which the hosts have been isolated. swimming cercariae age. In the laboratory, many of the cercariae fail Posterolateral locomotor organs are more to encyst and die within a few hours. Failure easily seen in developing cercariae than in the of encystment of many cercariae suggests the emerged forms. This is probably due to the premature emergence of some of the larvae development of pigmentation in the older due to laboratory conditions. Bioculate and forms; they are extremely important in any trioculate cercariae are found at the same time crawling movements. and from the same snail host, the trioculate Study of the development of the excretory being the more numerous. During develop- system indicate that it is in general agreement ment, the poorly organized third eyespot ap- with that described for the Family (Kuntz, pears after the more posterior two are well 1951). However, Figures 8, 11, 14, 15, and formed and when the cercariae are free in 16 show that there are two parts to the func- the host's tissue spaces. Emergence of the tional bladder. The double caudal duct, lead- bioculate forms also probably is due to un- ing from the atrium as seen in developing natural laboratory conditions. forms dissected from recliae or lymph spaces Large vesicular bodies, first described by of the snail host, cannot be observed by the

FIGS. 16—26. 16. Live, developing cercaria from snail. Collar, rudiments of reproductive organs, excretory system. (48x)- 17. Live, emerged bioculate cercaria. Digestive, excretory, and reproductive systems. (lOOx). 18. Live, emerged trioculate cercaria. Collar and all systems evident. (48x)- 19. Live, tail of young developing cercaria ex snail. Numerous vesicular bodies. (150x). 20. Live tail of older developing cercaria ex snail. Fewer vesicular bodies present. (150x)- 21. Live tail of developing cercaria. Arching caudal ducts, apparent mucoid secretions around tail. (150x). 22. Live cercaria beginning to encyst. First, thin irregular layer visible. (lOOx). 2-3. Live. Metacercaria approximately 50 days old. (150x). 24. Live metacercaria dissected ex cyst. Approximately 50 days old. (150x). 25. Live adult recovered from experimentally fed and laboratory-raised Chelonia my das. Ten days old. (150x). 26. Composite drawing of "mature" cercaria. Ventral view. Note primorclia of reproductive system as observed in sections.

FIGS. 27-29. 27. Composite sagittal section to show relationship of digestive, excretory, and primordia of reproductive systems. 28. Schematic representation of flame-cell pattern in "mature" cercaria. 29. Detailed relationship of cephalic glands, nervous system, and three eyespots in "mature" cercaria.

Copyright © 2011, The Helminthological Society of Washington JANUARY, 1967] HELMINTHOLOGICAL SOCIETY 39 time the cercariae emerge. The tubes in a opacity of the animal which was first described developing cercaria end blindly in the mid- for the adult. The number of flame cells is region of the tail (Fig. 7, 8). Aforementioned increasing over that of the cercaria, this is cystogenous cells and pigmentation make it evident by the spotting of an additional pair very difficult to see the smaller ducts and in the mid-lateral region. Pigmentation of the complete flame-cell pattern. The anterior part animals prevents following this development of the cycloid duct (Fig. 26, 28) is incon- through to include the adult. Worms have re- spicuous indeed in this larva and only careful mained viable within cysts kept in the lab- observations confirm its presence. Repeated oratory for a period of 5 to 6 months (Fig. 24). observations and plotting the location of flame The larvae decrease in size with age; the cells have definitely established the pattern average size of those dissected from month-old to be that ascribed to the Family by Kuntz, cysts measured 0.369 long and 0.166 wide. 1951. Measurements of the testes in a 42-day-old larva were 0.0208 by 0.0204 and 0.0194 by CYST FORMATION AND METACERCARIA 0.0218; overall size of this metacercaria was Cyst formation occurs very rapidly. The not determined because of damage during cercaria settles on a suitable substrate, con- dissection. tracts strongly, and secretes cystogenous ma- In unpublished data, Dr. John J. McDermott, terial from the entire body surface. While the Jr., found similar cercariae in Nassarius obso- secretion is taking place, the violently lashing leta from tidal ponds and marshes of the south- tail detaches from the body; often the posterior western coast of New Jersey. He noted that body region contracts to form a small rounded mature cercariae are often much larger than knob between the locomotor pockets, aiding the parent redia; this is true of my observa- in the tail detachment. The body of the cer- tions. The cercariae are also larger than the caria revolves actively, is contracted so as to metacercariae which tend to decrease in size be almost circular in outline, molding the cyst with the duration of encystment. McDermott wall during its movements (Fig. 22). The cyst also noted the primordia of the gonads (testes wall is formed in layers, the first secreted being and ovary), but did not see the entire cecae irregular and spread out on the substrate. The nor excretory system. In spite of the failure completed cyst is round, dome-shaped, and to see the complete details, such as collar, firmly attached. Although a thin wall is diverticula on gut, etc., I believe that he formed within 2 to 3 min, cyst wall forma- worked with the same larva; his measure- tion is not completed for approximately 8 to ments, comparisons of the general morphology, 12 min. A well-formed cyst wall averages behavior, method of encystment, the meta- 0.026 mm in thickness, and with age often cercaria, and the fact that many northern appears yellow-brown in color. The cyst diam- diamond-back terrapins were present in his eters exclusive of the attachment layer range collecting area permit me to name the New from 0.315 to 0.341, 10 averaging 0.330. The diameter of the space within the cyst averages Jersey coast as a second locality for Pleuro- 0.294 (Fig. 23). gonius malaclemys. When cyst formation is complete, the larva Detailed comparisons with other marine relaxes, may lie in a bent or folded position; monostomate cercariae, e.g., C. ephemera Le- or as it ages, may shorten and lie straight across bour, 1911, C. lebouri Stunkard, 1932, and the cavity. Older metacercariae are less active C. caribbea I Cable, 1956, are not being given. than the younger, and actually show little The forms can be compared only as to the development beyond that of the cercariae. The overall size and other general characteristics eyespots tend to lose their organization; rem- due to the difficulties of observing details nants of them are carried over into the adult which resulted in incomplete descriptions. C. worms as dark blotches of pigment. As would caribbea I is the only one definitely described be expected, the digestive system is much as a marine pronocephalid. Size, host differ- more visible than in the cercaria. Most of the ences, as well as Cable's observations on the internal organs arc obscured by the natural ecological relationships and probable adult

Copyright © 2011, The Helminthological Society of Washington PROCEEDINGS OF THE [VOL. 34, No. 1 affinities, allow me to take the liberty of Besides using an unnatural host, it is very assuming it to be a different species. probable that the changing physiological conditions naturally occurring in them played a FEEDING EXPERIMENTS significant role in the results. So far, it has Although morphology of both the cercaria been impossible to obtain laboratory-raised and metacercaria strongly indicate that the Malaclemys for the experiments. However, above described forms are larvae of Pleuro- the two successful feedings do confirm the gonius malaclemys Hunter, 1961, feeding ex- accurate identification (Fig. 25). periments were undertaken to corroborate the identification. Snail opercula heavily covered LITERATURE CITED with cysts were fed to the experimental hosts CABLE, R. M. 1956. Marine cercariae of Puerto used. Cysts on the opercula were checked for Rico. Sci. Survey of Puerto Rico and the viable metacercariae before feeding them. Virgin Islands XVI Part 4. N. Y. Aead. Sci- Three small and immature adults were ob- ences, April 1956. tained from the posterior region of the small FAUST, E. C. 1917. Notes on cercariae of the intestine of an adult female Malaclemys t. Bitter Root Valley, Montana. J. Parasit. 3: centrata 10 days after feeding. Forty-three 10,5-123. mature adult worms were obtained from the GENETZINSKAJA, T. A., AND A. A. DOBROVOLSKI. 1962. Glycogen and fat in various phases of same host, indicating that the experimental the life cycle and Trematocla. Leningrad Univ. infection was superimposed on a natural one Herald No. 9. Biol. Series, Part 2. even though the turtle had been kept in the HUNTER, WANDA S. 1961. Pleuragonius mala- laboratory for 3 months prior to the infection. clemys n. sp. (Trematoda: Pronocephalidae) All turtles kept in the laboratory were fed from Beaufort, North Carolina. Proc. Hel- only frozen shrimp. minthol. Soc. Wash. 28(2): 111-114. Through the courtesy of Dr. Peter Klopfer KRUIDENIER, F. J. 1953. The formation and of the Zoology Department of Duke University, function of mucoicls in cercariae: monostome eight 11-month-old Chelonia mydas which had cercariae. Trans. Amer. Micr. Soc. 72(1): been laboratory-raised and shrimp-fed were 57-67. made available to me. These turtles change , AND K. N. MEHRA. 1957. Mucosub- their feeding habits from carnivorous to herbiv- stances in plagiorchoicl and monostomate cer- orous during their second year and using them cariae (Trematoda: Digenea). Trans. 111. as experimental hosts was considered question- Acad. Sci. 50: 267-277. able. However, in one of these hosts, eight KUNTZ, R. E. 1951. Embryonic development of the excretory system in a psilostome cercaria, immature worms were recovered 10 days after a gymnocephalous (fasciolicl) cercaria and in cyst-feeding. Turtles which were kept 16 to three monostome cercariae. Trans. Amer. 24 days after feeding yielded no Pleurogonius. Micr. Soc. LXX (2): 95-118.

The Effect of Goiiadectomy and Hormone Therapy in Male Hamsters Upon the Egg Output by the Pseudophyllidean Diphyllobothrium sebago*

MARVIN C. MEYER AND WILLIAM G. VALLEAU

Mammalian sex hormones have been shown icine dropper; care was taken to place the experimentally to influence the burden of para- dropper well back in the oral cavity to avoid sitic helminths. The female hormone generally entry into the cheek pouches. Plerocercoids increases the host's resistance to infection, used in the infections were obtained from while the male hormone typically induces a viscera of landlocked salmon (Salmo salar) slight increase in the level of helminthiasis and brook trout (Salvelinus fontmalis) from (Haley, 1958; Mathies, 1959; Stahl, 1961; Mooselookmeguntic Lake. The pepsin—HC1 Dobson, 1962, 1964; Miller, 1965). digestion technique was used in larval re- Elimination of the endogenous hormone covery. Larvae were transferred to 0.75% through castration has also been shown to physiological saline solution, and nonmotile influence the worm. Gonadectomy of mice of larvae were discarded. Host animals were both sexes, infected with Aspiculuris tetraptera, maintained on an ad libitum diet of Purina significantly lowers the worm burden (Mathies, Laboratory Chow, supplemented with lettuce 1959). Solomon (1964) has shown that cas- leaves and water. Animals were housed indi- tration decreases the growth of Nippostrongy- vidually in 23- by 23- by 40-cin metal cages his brasiliensis in the male hamster, and that with wire mesh floors. Excelsior was used as this effect can be reversed by the administra- bedding. tion of testosterone. Seven days after the hamsters had been Similarly, Addis (1946), working with the infected, their feces were collected and ex- cyclophyllidean Hymenolepis diminuta in rats, amined for eggs to determine patency of the showed that castration of the male causes host. At 14 days postinfection the patent stunting in the worm and that the growth of hosts were divided into experimental groups the worm could be returned to normal in the and one-half were anesthetized with ether and emasculated animal by the administration of castrated. The following groups were estab- testosterone. The egg output of H. diminuta lished: (1) noncastrate untreated, (2) non- also was decreased in castrate male rats, but castrate treated, (3) castrate untreated, and could be restored to normal levels by the ad- (4) castrate treated. The administration of ministration of testosterone (Beck, 1952). testosterone began on the day following cas- These observations indicated that normal tration and was given at the 1-mg level in growth and normal egg output of H. diminuta 0.1-ml sesame oil per animal daily for 3 weeks. is directly related to the homologous sex Egg counts were made at 14, 21, and 35 hormone. days for each hamster by collecting the feces This report relates the effect of the male for a 24-hr period and soaking them over- gonadal hormone on egg output by a pseudo- night in a volumetric flask; the volume was phyllidean tapeworm, Diphyllobothrium se- then adjusted to either 100-ml or 250-ml, as bago, in male hamsters (Mesocricetus aiiratus). necessary, and the flask was shaken vigorously. A 0.1-ml sample was then withdrawn with a MATERIALS AND METHODS micropipette from near the center of the sus- Forty male hamsters, weighing from 80 gm pension and examined microscopically, the to 100 gm, from Dennen Animal Industries, cover glass (60 mm) having been sealed with Gloucester, Massachusetts, were experimentally vaseline to prevent drying and shifting of the infected with D. sebago larvae. Each animal eggs. The procedure was repeated to provide received three plerocercoids orally from a med- three egg counts and the mean was determined for each animal. The mean egg count for each * Department of Zoology, University of Maine, Orono, animal, multiplied by 1,000 for the 100-ml Maine. Supported by research grant (Al-0275) from the National Institutes of Health, U.S. Public Health Service. volume and by 2,500 for the 250-ml volume,

of the primary strobila, which, in gulls, occurs NONCASTRATE UNTREATED at about 2 weeks postinfection (Meyer and

CASTRATE Vik, 1963). The noncastrate untreated animals ITREATED showed an increase from 61,000 eggs at day 21 to 115,000 at day 35; while treatment of the noncastrate group increased the egg output from a value of 51,000 at 21 days to 128,000 at day 35. In the untreated castrate group, the number of eggs produced 21 days postinfection was 8,000. Eggs were not detectable in the feces at 35 days after infection, despite the fact that two hosts each harbored a gravid worm at necropsy. When testosterone was administered to castrate animals, not only was the 21-day decrease in egg output reversed, but the egg production level was highest in this group throughout the period. The egg count increased from 126,000 at 21 days to 138,000 at day 35 (Fig. 1). The wet weight of the spleen, kidneys, and adrenals did not vary significantly among the various groups. The decrease in the testes' weight of the noncastrate treated animals and the seminal vesicle stimulation is merely a reflection of effectiveness of the hormone FIG. 1. Egg output of Diphyllobothriiim sebago (Table 1). in noncastrate untreated and treated, and castrate The length, weight, and point of attachment untreated and treated male hamsters. The number of the worms did not vary significantly among of hosts in each group is indicated by the roman the groups. Two hamsters, one in each of the number adjacent to it. untreated groups, harbored two worms. Of the 18 positive hosts at 35 days postinfection, represented the egg output for the selected the remaining 16 each harbored one worm 24-hr period. The mean egg counts for the at necropsy. different periods were expressed to the nearest Although the data presented are not exten- thousand (Fig. 1). sive, they suggest that there is a relationship Animals were sacrificed 35 days postinfec- between D. sebago egg production and the tion by craniocervical dislocation. At necropsy male sex hormone in hamsters. These obser- the intestine from pylorus to cecum was excised vations strengthen the conclusions of earlier and placed atop a ruler divided into units of workers (Addis, Beck, Mathies, Solomon) that 1 mm. The ruler provided quick measurement the endogenous male sex hormone is related to of both the intestinal length and the site of helminthiasis. scolex attachment. The worms were then transferred to saline solution, washed free of extra- SUMMARY neous materials, and dried between paper The suggestion that the male sex hormone towels; they were then measured to the near- influences the egg output by Diphyllobothriiim est mm in length, and weighed on a torsion sebago in hamsters was examined. Fourteen balance to the nearest mg. days postinfection with plerocercoids one-half of the patent hosts were castrated. Four ex- RESULTS AND DISCUSSION perimental groups were established: (1) non- All groups showed an initial decline in egg castrate untreated, (2) noncastrate treated, production at 21 days postinfection. Possibly (3) castrate untreated, and (4) castrate this decrease in egg output coincides with loss treated. Testosterone administration began on

TABLE 1. Average weights (mg) of organs from noncastrate untreated and treated, and castrate untreated and treated hamsters at necropsy. The number of animals is indicated in parentheses for each group.

Group Treatment Adrenals Spleen Kidney Seminal vesicle Testes Noncastrate (4) 20.1 66.6 909.8 309.8 1,604.0 Noncastrate (5) Testosterone, 1 mg X 21 days 18.5 64.8 939.5 555.7 384.0 Castrate (2) 18.2 64.7 824.6 42.7 Castrate (7) Testosterone, 1 mg X 21 days 17.6 64.0 816.6 772.1 —

the clay following castration and was given nadectomy, and thyroidectomy on experimen- at the 1-mg level in 0.1-ml sesame oil per tal infections in lambs. Exp. Parasit. 15: 200- animal daily for 3 weeks. Egg counts were 213. made at 14, 21, and 35 days postinfection. The HALEY, A. J. 1958. Sex difference in the resistance of hamsters to infection with the rat castrates showed a decline in egg production nematode, Nippostrongylus mum. Exp. Para- throughout the 35-day experimental period, sit. 7: 338-348. this effect was reversed by the administration MATHIES, A. W. 1959. Certain aspects of the of the male hormone. host-parasite relationship of Aspiculuris tetraptera, a mouse pinworm. II. Sex resistance. LITERATURE CITED Exp. Parasit. 8: 39-45. ADDIS, C. J. 1946. Experiments on the relation MEYER, M. C., AND R. VIK. 1963. The life cycle between sex hormones and the growth of of Diphyllobothrium sebago (Ward, 1910). J. Parasit. 49: 962-968. tapeworms (Hymenolepis diminuta) in rats. J. Parasit. 32: 574-580. MILLER, T. A. 1965. Influence of age and sex BECK, J. W. 1952. Effect of gonadectomy and on the susceptibility of dogs to primary infec- gonadal hormones on singly established Hy- tion with Anci/lostoma caninum. ]. Parasit. 51: 701-704. menolepis diminuta in rats. Exp. Parasit. 1: SOLOMON, G. B. 1964. Effect of gonadectomy 109-117. and hormone therapy on Nippostrongylus bra- DOBSON, C. 1962. Certain aspects of the host- siliensis (Travassos, 1914) infections in male parasite relationship of Nematospiroides hamsters. J. Parasit. 50 (3, Sec. 2): 57. dubhis (Baylis). V. Host Specificity. Para- STAHL, W. 1961. Influences of age and sex on sitology 52: 41-48. the susceptibility of albino mice to infection . 1964. Host endocrine interactions with with Aspiculuris tetraptera. J. Parasit. 47: nematode infections. I. Effects of sex, go- 939-941.

Studies oil the Life History of Two Notocotylids (Trematoda)

ALEXANDER D. ACHOLONU* AND O. WILFORD OLSEN Department of Zoology, Colorado State University, Fort Collins, Colorado

INTRODUCTION The status of certain species of Notocotylus Certain aspects of the life history of Noto- has been revealed by Szidat, 1936; Harwood, cotijlus stagnicolae Herber, 1942, and Noto- 1939; Ruiz, 1946; and Dubois, 1951. cotylus urbanensis (Cort, 1914) worked out in Cort (1914) described a monostome cer- Fort Collins, Colorado, reveal new information. caria as Cercaria urbanensis. Harrah (1922) postulated the life history of Notocotylus ::: Present address: Department of Biology, Southern Uni- versity, Baton Rouge, Louisiana. urbanensis on the basis of morphological com- A portion of a dissertation submitted to the Graduate parison of encysted cercariae and young adults School, Colorado State University, in partial fulfillment of the requirements for the degree of Doctor of Philosophy. he found infecting muskrats, Ondatra zibe- This investigation was supported by a grant from the Federal Republic of Nigeria. thicus (L.), a wood duck, Aix sponsa (L.),

Copyright © 2011, The Helminthological Society of Washington 44 PROCEEDINGS OF THE [Voi,. 34, No. 1 and a pintail duck, Anas acuta (L.). Lutter- and host records of the two monostomes is moser (1935) published an account of the given. life history study of N. urbanensis which supported Harrah's work. In this paper, he re- MATERIALS AND METHODS ported obtaining immature adult Notocotylus Natural infection by two different species urbanensis with undeveloped ventral cuticular of trioculate monostome cercariae was found glands from a muskrat 11 days post-feeding in the snails, Lymnaea auricularia (L.) col- and mature adults containing ova from a duck- lected from Dixon Lake near Fort Collins, ling 25 days after infection. He, however, Colorado, and Physa gyrina (Say) from var- expressed some doubt as to the validity of his ious ponds and irrigation canals in the Fort identification of the immature parasites since Collins area. they lacked the ventral glands necessary for Mature cercariae were studied alive both distinguishing Notocotylus urbanensis from unstained and stained with mtravitam dyes of Quinqueserialis quinqueserialis Baker and Neutral Red and Nile Blue Sulfate. Photo- Laughlin, 1911. Berber (1942), who worked micrographs were made of some living cer- out the life history of Q. quinqueserialis, cariae when possible. Cercariae were fixed in stated that this parasite attains maturity in hot 10% formalin. Permanent whole mounts about 2 weeks. Since Luttermoser's speci- were studied after staining with either Mayer's mens were still immature and lacked ventral acid carmine or Harris' alum hematoxylin. glands after 11 clays, it is apparent that they Cysts formed by emerged cercariae were also were not Q. quinqueserialis but most likely studied alive and as permanent whole mounts N. urbanensis. Harwood (1939) questioned stained in Mayer's acid carmine. The speci- the validity of Harrah's (1922) work and stated mens were cleared in beechwood creosote and that "the adult form of Cercaria urbanensis mounted in piccolyte. Measurements of pre- Cort (1914) has never been properly identi- served material, unless otherwise specified, rep- fied, and the specific name urbanensis Cort, is resent the average for five to 10 well extended without validity for adult trematocles until the or fairly well extended specimens. The maxi- connection has been experimentally demon- mum, minimum, and average for the speci- strated." In the same year, Herber (1939) mens were recorded, with the average being reported recovering mature adult N. urbanensis in parentheses. Measurements of all specimens from a muskrat on the 10th day after infection. were made with the aid of an ocular microm- Later, he (1955) published a fuller account eter and are in microns. Controlled infection of the life history of this parasite in which he, experiments were conducted to obtain infor- like Harwood (1939) discredited Harrah's mation on the sexually mature stages and con- work on the premise that Harrah "did not firm the larval identifications. Cysts formed have material which developed from Cercaria by cercariae of N. urbanensis (Fig. 3) on the urbanensis." He listed Catatropis fimbriata inside wall of containers holding Physa gyrina were collected in stender dishes and force-fed Baker, 1915 (= Catatropis filamentis Baker, by means of pipette or medicine dropper to 1915) and Paramonostromum echinum Harrah, four laboratory-reared 1- to 4-day-old chicks. 1922 as being conspecific with N. urbanensis. These chicks were fed 125, 200, and 300 cysts, No information was given on the prepatent respectively. Also, cysts formed from cercariae period of N. urbanensis, especially in the musk- of N. stagnicolae which emerged from Lym- rat, the natural host which was used as one naea auricularia (L.) were, together with of his experimental animals. whole infected snails, fed to laboratory-reared It is the purpose of this paper to report the 19- to 35-day-old ducklings, 13-day-olcl gos- study made on the life history of Notocotylus lings, 19- to 35-day-old chicks, and 9-week-old stagnicolae and N. urbanensis. In addition, an albino rats. All infection experiments were attempt was made to resolve the conflicting conducted with cysts less than 1 day old, and statements made by Luttermoser and Herber our experimental animals were autopsiecl be- concerning the time of maturity of N. urban- tween 11 and 14 days after infection. Adult ensis. New information regarding distribution worms recovered were studied alive as well

TABLE 1. Comparison of 5-clay-old N. stagnicolae and N. urbanensis (measurements in microns).

N. stagnicolae N. urbanensis

Length of body 902-1,243 (1,036.75) 451-605 (528) Width of body 209-297 (223.5) 84-127.2 (103.6) Oral sucker Length 55.2-64.8 (58.6) 38.4-45.6 (42) Width 60-72 (65.2) 38.4-45.6 (41.2) Length of esophagus 50.4-96 (73.2) Obscured by pigment; about 36—47 ( 42 ) Ventral glands Rudimentary; faintly seen Undeveloped No. in lateral row About 11-12 No. in middle row About 13 Tcstes No. of lobes, lateral margin 5-6 Not well defined Length m l fi^ / 1 TO 9 "1 Width 60 79 9 ( 67 9 ) Distance from posterior end oo c ,10 / /to c \2 (63.2) Ovary Length Not well defined Width ( greatest part ) 40.8-60 (49.96) Vitellaria Developed but not well defined; Undeveloped mass of small cells Uterine loops between vitellaria and Not well defined; wavy and more Not well defined; rudimentary cirrus sac advanced than in N. urbanensis Pigmentation Sparse and scattered Heavily pigmented especially at the anterior part

as when fixed in 10% formalin and stained become infected, while one harbored a single in Mayer's acid carmine. parasite. A comparison was made between 5-day-old N. urbanensis obtained from a chick and N. DISCUSSION stagnicolae of the same age recovered from Since the work of Harrah (1922) on the a duckling (See Table 1, Figs. 1, 2). North American monostomes, knowledge of species of Notocotylus has been augmented RESULTS greatly. Five days postexposure, the chick fed over It is interesting to note that N. urbanensis, 300 metacercariae of N. urbanensis died. When a natural parasite of muskrats, developed in autopsiecl, 13 preadult worms (Fig. 1) were a chick. Herber (1939) also reported obtain- recovered from the large intestine. On the ing this worm from a rooster. Luttermoser 14th day after exposure, the remaining chicks (1935) recovered it from ducklings and were autopsiecl and found to be free of trem- Kuntz (1951) from Microtus pennsylvanicus atodes. (Orel, 1815). In this investigation, N. stagni- Mature N. stagnicolae were obtained from colae, which is a parasite of ducks, developed the large intestine and ceca of the experi- to adulthood in chicks, ducklings, goslings, mentally infected animals. The ceca were, and an albino rat. These results unequivocally however, more heavily infected. The first suggest that while the adult species of Noto- infection was found in a duckling autopsied cotylus may have preferential hosts, they are 5 clays after administering metacercariae. non-host specific. This is a fact substantiated Twenty-one preadult worms (Fig. 2) with by Stunkard (1960) who pointed out that rudimentary cuticular ventral glands were "host specificity of the adult stages is not found. Of seven chicks exposed to infection, definite, because the same metacercariae can five were positive. The largest number of mature in a number of different host species." worms collected from one chick was 34. Two In this respect, Herber (1942) also observed of three ducklings became infected. One duck- that certain of the earlier authors considered ling harbored 83 adult worms. Two goslings adults found in different hosts as separate were both positive. One was infected with species. This, according to him, is obviously 38 worms. Three of four albino rats did not unsound as one species may develop in a wide

n O O ro

FIGS. 1-2. 1. Five-day-old Notocotylus urbanensis. Dorsal view. 2. Five-day-old Notocotylus stagnicolae. Dorsal view.

Copyright © 2011, The Helminthological Society of Washington JANUARY, 1967] HELMINTHOLOGICAL SOCIETY 47 variety of animals. He cited N. stagnicolae as an example of a parasite of birds that developed to maturity in mammals. Szidat and Szidat (.1933), on the other hand, postulated that specificity is very marked in the case of the snail hosts. This view was supported by Herber (1942), who added that although the degree of this specificity is not known at present, it probably has real significance. The specificity of N. urbanensis or N. stagnicolae to different species of snail hosts is difficult to evaluate since it was not until 1955 that Herber demonstrated that the two species are different. In view of the close similarity of Cercaria urbanensis and Cercaria stagnicolae, all previous host records of C. urbanensis must be regarded with caution. However, if one assumes no misidentification by previous authors, the larval stages of the two species of Notocotylus show little host specificity. Cort (1914) reported finding Cercaria urbanensis in Physa gijrina; McCormick (1923) in Gonio- basis livescens (Menke); Luttermoser (1935) in Stagnicola emarginata angulata (Sowerby), Physella parkeri (Currier), and P. magnala- custris (Walker, 1901); Herber (1939) in several species of Physa and Stagnicola emarginata angulata. Cercariae of N. stagnicolae were obtained from Stagnicola emarginata FIG. 3. Photomicrograph of living cercaria of angulata. (Sowerby) and S. e. canadensis Notocotylus urbanensis under cover slip pressure. Shows the three eyespots, the excretory bladder, (Sowerby) by Herber (1942); from Stagnicola and the excretory tubules joining above the median palustris (Mtiller) by Wu (1953); and from eyespot. Lymnaea auricularki (L.) by the present authors. The infection experiments show that al- the same age. While there is incipient forma- though N. stagnicolae is non-host specific, the tion of the ventral cuticular glands in N. adults develop more readily in birds than in stagnicolae, there is no evidence of this in mammals. Of four albino rats fed metacer- N. urbanensis. The reproductive organs are cariae, only one was infected, and with a fairly well developed in N. stagnicolae but single adult parasite recovered, while each of undeveloped or not well defined in N. urban- the bird species used for experimental infec- ensis (Table 1, Figs. 1, 2). The heavy pig- tion sustained a fairly large number of worms. mentation which is characteristic of the cer- Herber (1942) fed cysts to five rats, two cariae and metacercariae of N. urbanensis is meadow mice, and one white mouse but re- still noticeable in this worm, especially at the covered only one immature specimen from the anterior end, while it is sparse and scattered white mouse and four worms from two rats. in N. stagnicolae. From this it is evident that Wu (1953) got negative results from experi- N. stagnicolae at 5 days is far more advanced mental infection of two white mice and two developmentally than N. urbanensis of the hamsters. same age. The fact that these worms came Table 1 shows that the preadult N. stagni- from two different but related species of birds colae (L. 1,036 p., W. 223.5 ^) is much larger (chick and duckling) probably made little or than N. urbanensis (L. 528 /.,„ W. 103.6 M) of no difference. The parasites are non-host spe-

TABLE 2. Comparison of three Notocotylids (measurements in microns,

N. triserialis N. urbanensis IV. stagnicolae Diesing, 1859 (Cort, 1914) Herber, 1942 ( after Dubois, 1951) Herber, 1955 Length of body 1,500-2,640 2,670-3,620 2,670-3,400 (3, 110 ±220) ( 2,990 ± 180) Width of body 480-700 730-920 720-940 ( 800 ± 60 ) (830 ±60) Oral sucker ? 110-130 130-180 (120 ±50) (160 ±8) Length of esophagus 90-145 ? 90-180 Ventral glands No. in lateral row 14-16 15-19 (17) 14-17 (15) No. in median row 14-15 13-18 (16) 13-15 (14) Testes No. of lobes 2-4 8-12 Length •350-360 270-350 340-560 ( 320 ± 20 ) ( 440 ± 40 ) Width 170-210 170-230 170-300 (200 ±20) ( 240 ± 20 ) Distance of testes from ? 130-310 60-140 posterior end (210±40) ( 100 ± 60 ) Ovary Length 170-180 160-210 200-300 (180± 10) ( 230 ± 20 ) Width • 130-210 210-300 (150 ±20) (260 ±20) Vitellaria 17—27 groups 24-34 groups of of follicles not follicles on each reaching to side middle of body Position Extend up to 2.1-2.5 (2.3) 1.8-2.1 (1.9) 0.52-0.57 of length of worm (0.53) Uterine loops between 3-5 9-13 (11) 3-5 (4) vitellaria and cirrus sac on metraterm side cific and previous records show that matured ever, he thinks that they were from natural adults of both have been recovered from either infections. Based on recent additional work of the hosts. According to Herber (1942), N. on this species, he (Herber, 1964) added in stagnicolae develops to maturity in about 10 his communication that N. urbanensis matures days. He (1939) recovered mature N. urban- in laboratory rats and Microtus in about 24 ensis 10 days after infection. On the basis days, which is a day shorter than the time of the present study, it seems highly improb- reported by Luttermoser (1935). This state- able for N. stagnicolae and N. urbanensis to ment is apparently more accurate than that of attain maturity in the same length of time. 1939 and verifies the work of Luttermoser. Moreover, it was pointed out by Acholonu Dubois (1951) considered N. urbanensis (1964) that our specimens and those of Lutter- (Cort, 1914), Harrah (1922) ex parte and moser (1935) agree somewhat in their rate of N. stagnicolae Herber, 1942, as synonyms of growth. It would appear, therefore, that the zY. triserialis Diesing, 1839. Available informa- development of N. urbanensis to sexual mation shows that this synonomy is not valid. turity may be slower than reported by Herber 1. Dubois' (1951) synonymy is based on (1939), who stated (pers. comm.) that at the the N. urbanensis collected by A. Hassall in time he thought the specimens from the musk- Maryland, USNM: No. 5772, No. 5771, No. rats were the result of experimental infection, 5769, and No. 5770, upon which Harrah's and that he is no longer certain about them. (1922) work was based and about which He had kept the muskrat for a few weeks and Herber (1955) wrote ". . . it seems justifiable then fed it some cysts. When the muskrat to conclude that Harrah did not have material died, he found the specimens which he re- which developed from Cercaria urbanensis, ported to have matured in 10 days; now, how- and that, therefore, his description of Noto-

TABLE 3. Comparison of the Larval forms of three Notocotylids (measurements in mierons). Cercaria of N. Cercaria naga urbanensis Cercaria of N. Szidat and (Con, 1914) stagnicolae Szidat, 1933 (after Herber, 1955) Herber, 1942 Body of cercaria Length 544 351-406 (383) 320-570 Width 152 140-187 (161) 80-210 Tail 600 671-764 (721) 620-940 Excretory tubules Join anteriorly eaudad Join anteriorly ahead Join anteriorly eaudad of median eyespot to median eyespot Time of encystment p Very soon after emergence; Delayed: 5-20 min usually within 3—5 min Cyst size 230 220-260 200-250 Mother sporocyst p Contain 4—8 mother redia Contain only one mother redia Mother/ redia Only daughter redia figured 308-871/81-308 170-1,330/80-190 but measurement not given cotylus urbanensis must be discarded." Har- and different species (see Tables 2 and 3). wood (1939) previously expressed this same While this comparison brings out some sim- notion. He stated that "the adult form of ilarities in these three species, in our opinion, Cercaria urbanensis Cort, 1914 is without the dissimilarities are enough to warrant their validity for adult trematodes until the connec- consideration as separate species. We are in tion has been experimentally demonstrated." agreement with Herber (1955), who gave the Herber (1955) (4 years after Dubois' work) following diagnostic characters as being helpful experimentally demonstrated this connection in separating species of the genus Notocotylus: and thus established N. urbanensis as a sep- (1) the anterior extent of the vitellaria, (2) arate and valid species. the lateral lobation of the testes, (3) body 2. The life history of N. stagnicolae and and tail size of cercariae, (4) the course of N. urbanensis conducted by Herber (1942, the excretory tubules of the cercariae, and 1955) and restudied by the present authors (5) cyst size. Tables 2 and 3 show that these shows that these two notocotylids are not worms differ, inter alia, with respect to num- identical (see Table 1). Also, a comparative bers 1, 2, and 4. examination of these two and the type species, Stunkard (1960) stated that the determina- N. triserialis, as described by Dubois (1951), tion of species of Notocotylus is difficult. The reveals that the three are seemingly separate worms, according to him, are very similar and

TABLE 4. Species of Notocotylus with their life history worked out. Species Cercaria Intermediate host (s) 1. N. stagnicolae Herber, 1942 Cercaria of N. stagnicolae Herber Generally in Stagnicolae spp.; 1942 Lymanaea auricularia (L.) 2. N. urbanensis (Cort, 1914), Cercaria urbanensis Cort, 1914 Generally in Phyaa spp. Goniobani.i Herber, 1955 livescens (Menke), Stagnicola Syn. Paramonostomum echinnm emaginatti iingulata ( Sowerby ) Harrah, 1922; Catatrnpi.i fimbriata Barker, 1915 C. filamentis Barker, 1915 N. triserialis Diesing, 1839 Cercaria vaga^ Szidat and Szidat, Lymnaea pahistris (Mull.) 1933 N. ephemera (Nitzsch, 1807) Cercaria ephemera Nitzsch, 1807 Planorbis corneus ( L.) (= N. thienemanni) Szidat and Szidat, 1933 5. N. imbricatus Szidat, 1934 Cercaria imbricata Looss, 1893 Bithynia tentaciilata (L.) 6. N. minutus Stunkard, 1960 Cercaria of N. minutus Hydrobia miniita (Totten) Stunkard, 1960 7. N. seineti Fuhrmann, 1919 Cercaria monostomi Looss? (after Szidat, 1936) Described as the cercaria of IV. attemiatiis Rud. 1809 by Szidat and Szidat, 1933. (See Dubois, 1951: 54.)

Copyright © 2011, The Helminthological Society of Washington 50 PROCEEDINGS OF THE [VOL. 34, No. 1 specific differences in morphology are small. HARRAH, E. C. 1922. North American mono- Such being the case, the separation of closely stomes primarily from fresh water host. 111. related species and the establishment of syn- Biol. Monog. 7: 219-328. onymous ones (which so far is a point of much HARWOOD, P. D. 1939. Notes on Tennessee confusion) could better be accomplished helminths IV. North American trematodes of the subfamily Notocotylinae. T- Tenn. Aead. through life history studies. Of all known Sc. 14: 421-437. species of Notocotylus (20 listed by Dubois HERBER, E. C. 1939. Life history studies on (1951)) plus N. stagnicolae Herber, 1942, monostomes of the genus Notocotylus (Trem- N. urbanensis (Cort, 1914) Herber, 1955, and atoda) J. Parasit. 26 (Suppl.): 18. N. minutus Stunkard, 1960, only six or seven . 1942. Life history studies on two trem- have their life cycles elucidated (see Table 4). atodes of the sub-family Notocotylinae. }. Thus, the necessity of resolving the life history Parasit. 28: 267-275. . 1955. Life history studies on Noto- of more of the described species of the genus cotylus urbanensis (Trematoda: Notocoty- Notocotylus cannot be overemphasized. linae). Proc. Penn. Ac-ad. Sc. 29: 267-275. As far as the authors are aware, this is the 1964. Methods of differentiating trem- first record of N. urbanensis and N. stagnicolae atodes of the genus Notocotylus. Proc. Penn. from Colorado. The finding of N. stagnicolae Acacl. Sc. 38: 12. in Lymnaea auricularia (L.) represents a new KUNTZ, R. E. 1951. Embryonic development of host record. The goose is also a new experi- the excretory system in a psilostome cercaria, mental definitive host for N. stagnicolae. a gymnocephalous (fasciolid) cercaria and in three monostome cercariae. Tr. Am. Micr. SUMMARY Soc. 70: 95-118. LUTTEHMOSER, G. W. 1935. A note on the life A study was made of certain aspects of the history of the monostome, Notocotylus urban- life history of Notocotylus stagnicolae Herber, ensis. J. Parasit. 21: 456. 1942 and IV. urbanensis (Cort, 1914). McCoRMiCK, R. N. 1923. The comparative para- An attempt was made to resolve the con- sitization of certain fresh water snails. Ohio flicting statements made by Luttermoser (1935) J. Sc. 23: 163-180. Ruiz, J. M. 1946. Pronocephalidae (Tn-m- and Herber (1939) concerning the time of atoda). Estudos das especies Brasileras e re- maturity of N. urbanensis. This study backed viso da famila. Mem. Inst. Bntantan (Brazil) by Herber's (1964) work on N. urbanensis, 19: 249-372. and his personal communication, verifies Lut- STUNKARD, H. W. I960. Studies on the mor- termoser's work. phology and life history of Notocotylus mi- Available information from life history stud- nutus n. sp., a digenetic trematode from ducks. ies shows that N. urbanensis and N. stagnicolae, J. Parasit. 46: 803-809. considered as synonyms of N. triserialis, Die- SZIDAT, L., AND U. SZIDAT. 1933. Bcitrage zur sing, 1839, by Dubois (1951), are separate Kenntnis der Trematoden der Monostomiden- and valid species. gattung Notocotylus Diesing. Zentralbl. Bak- teriol. 1. Abt. Orig. 129: 411-422. LITERATURE CITED . 1936. Beitrage zur Kenntnis der Trem- atodengattung Notocotylus Diesing, 11. No- ACHOLONU, A. D. 1964. Life history of two notocotyliclae. J. Parasit. 50: 28-29. tocotylus linearis (Rud. 1819?) n. sp. aus CORT, W. W. 1914. Larval trematodes from den Blinddarmen des Kiebitz (Vanellm vanel- North American freshwater snails. J. Parasit. lus L.). Zentralbl. Bakteriol. 1. Abt. Orig. 1: 65-84. 136: 231-235. DUBOIS, G. 1951. Etude des trematodes Nord- Wu, L.-Y. 1953. On the life history and biology Americans cle la collection E. L. Schiller et of Notocotylus stagnicolae Herber, 1942 (Fam- revision clu genre Notocoti/lus Diesing, 1839. ily Notocotyliclae). Canacl. 1. Zool. 31: 522- Biol. Sc. Neuchatel, Sc. Nat. 74: 41-76. 527.

Paraphelenchus acontioides n. sp. (Nematoda: Parapheleiichidae), a Mycophagous Neinatode from Illinois, with Observations on its Feeding Habits and a Key to the Species of Paraphelenchus

D. P. TAYLOR AND J. K. PILLAI*

An undescribed species of the genus Para- nounced on the ventral side. Procorpus slender phelenchus (Micoletzky, 1922) Micoletzky, with conspicuous lumen. Metacorpus large, 1925 was recovered from soil collected around aphelenchoid, with valvular apparatus slightly the roots of Kentucky blue grass (Agrostis posterior to the center. Esophageal gland ducts palustris Huds.) on the Horticulture Research open into metacorpal lumen in typically aphe- Farm at the University of Illinois, Urbana. lenchoid arrangement. Length of the lumen This species was established in 1960 on between the openings of the glands and the the fungus Pyrenochaeta terrestris (Hansen) beginning of the valve is highly sclerotized. Gorenz., J. C. Walker, and Larson, and has Isthmus slender, gradually widening to form been maintained in the laboratory since that basal bulb. Three esophageal glands confined time. within basal bulb. Lumen of isthmus and basal bulb less sclerotized than that of corpus. In- Paraphelenchus acontioides n. sp. testine composed of two rows of easily discern- DIMENSION: Females (n = 60): L — 0.77 ible cells, visually filled with refractive globules ±0.04 mm (0.71-0.88 mm); a = 28.5 ± 1.4 in specimens from active cultures. Intestinal (25-31); b = 4.8 ±0.2 (4.4-5.3); c = 25 ± lumen forming a wide anterior chamber, be- 1.5 (20-30); V = 74.8 ±0.7 (73-77); Stylet coming narrower and sinuous throughout the = 15 ± 1 p (14-16 p.). rest of its length. Rectum about one and one- DESCRIPTION: Body slightly arcuate ven- half times the anal body diameter. Anal open- trally when heat-relaxed; gradually tapers an- ing about a third the body diameter at that teriorly to a low rounded lip region which is point as in Figure IF. Nerve ring surrounding continuous with the body contour. Cuticle the isthmus immediately behind metacorpus. marked by fine transverse striae, approximately Dierids papillate, conspicuous, located in the 1 /ji apart. Lateral field occupies one-fourth to lateral field at the level of the nerve ring. one-fifth body width and contains eight incis- Excretory pore opposite nerve ring with canal ures throughout most of its length. Number of leading ventrally to a unicellular gland ventral incisures reduced to six in the isthmus region, to the junction of esophagus and intestine. lateral field fading out anterior to corpus. Hemizonid just posterior to the excretory pore; Incisures also reduced to six in anal region, hemizonion one body width posterior to it. becoming indistinct near the tail tip. Body Phasmids located near the tail terminus ap- tapers posteriorly to a short tail, less than twice pearing as minute papillae each with a fine anal body diameter. Dorsal surface of the tail strand connected to the body content. curved more than the rather straight ventral Ovary single, outstretched. Oocytes ar- surface. Tail tip with a single mucro as in ranged in a single row surrounded by a con- Figure 1 F, G, and H. spicuous epithelium. Epithelial cells with Lip region not striated, consisting of six lips. nuclei slightly larger than in oocytes. Anterior Neither amphids nor papillae seen. Spear guid- portion of epithelium illustrated in Figure 1A; ing ring a circumoral circle composed of six posterior portion omitted for clarity. Uterus short pieces fused together (Fig. 1C). Stylet composed of paired ovoid cells ranging from 14—16 ^ long with a shorter conical anterior 7—13 in number. Post vulvar sac about twice portion and a cylindrical shaft. Stylet has the vulval body width in length. Vulva a slight basal swellings which are more pro- transverse slit with conspicuous vulvar lips, * Associate Professor and Research Associate, Depart- observable in young females prior to escape ment of Plant Pathology, University of Illinois, Urbana, Illinois, USA. from fourth stage cuticle. One or two eggs

05mm , BDEFGH

Copyright © 2011, The Helminthological Society of Washington JANUARY, 1967] HELMINTHOLOGICAL SOCIETY 53 may be present at a time in egg-laying females. 3. Lateral field with 6 lines, lip region con- Eggs laid in the single celled stage, measuring tinuous with the body contour 64-74 ,,, in length and 28-32 M in width. P. amblyurus Steiner, 1934 HOLOTYPE: Female, progeny of a female 4. Lateral field with 8 lines, lip region collected in 1960 and maintained on cultures continuous with the body contour of PyrenocJiaeta terrestris growing on Potato P. acontioides n. sp. Dextrose Agar, slide # P-H-1, Nematode Slide 4. Lateral field with 9 lines, lip region Collection, Department of Plant Pathology, offset P. pseudoparietinus University of Illinois, Urbana. (Micoletzky, 1922) Micoletsky, 1925 PARATYPES: Ten females, date same as for holotype, slide # P-P-1, Nematode Slide Col- 5. Spicules of males with a sharp ventral lection, Department of Plant Pathology, Uni- process, manubrium more than half the versity of Illinois, Urbana. length of the shaft 6 TYPE HABITAT AND LOCALITY: Soil around 5. Spicules of males without a ventral roots of Kentucky blue grass (Agrostis palustris process, manubrium less than half the Huds.), Horticulture Research Farm, Univer- length of the shaft sity of Illinois, Urbana. P. batavicus Filipjev, 1934 DIAGNOSIS: Paraphelenchus acontioides n. sp. 6. Female tail with a pair of subter- can be distinguished by the presence of a minal papillae, lip region continuous mucro on the tail tip from P. batavicus Filipjev, with the body contour 1934, P. myceliophthorus J. B. Goodey, 1958, „ P. myceliophthorus J. B. Goodey, 1958 and P. tritici Baranovskaya, 1958, all of which 6. Female tail without any tail process, lack such a terminal process. Each of those lip region offset species possessing mucrones has a character- P. tritici Baranovskaya, 1958 istic number of lines in the lateral field: P. basili Das, 1960 has four; P. amblyurus Steiner, FEEDING HABITS OF P. acontioides 1934 has six; nine were illustrated in P. pseudoparietinus Micoletzky, 1922; whereas P. acon- Observations were made using nematodes tioides n. sp. has eight. The new species also which had been transferred from stock cul- differs from P. pseudoparietinus, its closest tures to 4-day-old fungus cultures growing relative, in that the lip region in the former is in a very thin layer of 2% water agar in petri continuous with the body contour and males dishes. After a cover slip was placed over are not known, whereas the lip region in the them it was possible to observe details with latter is offset and males are known. a 90X oil immersion lens. Feeding habits described here are based on observations made KEY TO THE SPECIES OF THE while the nematodes were feeding on the GENUS Paraphelenchus fungus, Pyrenochaeta terrestris. This nematode is a voracious feeder, empty- 1. Tail tip with one or more mucrones 2 ing a cell within a few seconds and then 1. Tail tip without mucro 5 moving to the next cell in a very short time. 2. Tail tip tapering evenly dorsally and The same general area of feeding was some- ventrally to a blunt terminus, lateral times vised for more than an hour during which field with 4-6 lines 3 the posterior half of the body remained at the 2. Tail tip with prominent curvature same place, with only occasional movements, on the dorsal surface, lateral field while the anterior half moved in various direc- with 8-9 lines 4 tions to feed on cells within its reach. 3. Lateral field with 4 lines, lip region Movement of P. acontioides was of the usual offset P. basili Das, 1960 type with the head turning to the sides accom-

FIG. 1. Paraphelenchus acontioides n. sp., A, Female; B, anterior region; C, en face view; D, lateral field near the isthmus; E, lateral field near center of body; F, ventral view of tail; G and H, lateral view of tail tip, H-typical, and G-a variant.

Copyright © 2011, The Helminthological Society of Washington 54 PROCEEDINGS OF THE [VOL. 34, No. 1 panied by occasional protrusion of the stylet. parison and it differed in the following details. Once the head touched a hypha, the nematode A. avenae usually required about 20—50 stylet stopped and fed on it, or bypassed it. When thrusts for penetration of a cell wall, whereas a cell was chosen for feeding the nematode P. acontioides required only 10-15 stylet thrust its stylet into the cell with the head thrusts. The twitching movement associated appressed to the cell wall. The number of with the stylet action was in the upper quarter stylet thrusts usually varied from 10-15 at a of the median bulb in A. avenae, whereas P. rate of 15—20 thrusts per second; but in some acontioides exhibited a twitching movement cases penetration was achieved with only 4-8 at the esophago-intestinal junction and not at stylet thrusts. Number of stylet thrusts re- the median bulb. As a rule P. acontioides quired for penetration could not be correlated withdrew all the cell content from the cell, with the proximity of the septum, nature of whereas A. avenae frequently left the cell par- the cell wall, or the turgidity of the cell. The tially emptied. In A. avenae the cell contents action of the stylet was always associated with appeared as globules when pumped into the a twitching movement at the esophagus-intes- intestinal lumen, and not as a jelly-like cyto- tine junction. This movement was also ob- plasmic mass. The feeding habits of both the served during the metacorpal pumping and esophago-intestinal valve action. Once the species were similar in other details. stylet gained entry into the cell there was a SUMMARY short pause, lasting only a fraction of a second, and soon the metacorpal pump action started. Paraphelenchus acontioides n. sp., recovered After five to eight pumpings at a rate of about from soil samples around the roots of Kentucky four per second the nematode left the cell. As blue grass (Agrostis palusiris Huds.) is de- a rule this nematode withdrew all the cell con- scribed, its feeding habits compared with those tents. When pumped into the intestine the cell of Aphelenchus avenae, and a key to the spe- contents formed a jelly-like mass that was re- cies of the genus Paraphelenchus is given. tained in the transparent anterior chamber until the next feeding when it was pushed down LITERATURE CITED as such into the lumen. As the food passed BARANOVSKAYA, I. A. 1958. Kpoznaniyu roda along the lumen of the intestine it decreased in Paraphelenchus (Micoletzky, 1922) Micolet- bulk suggesting digestion and assimilation were zky, 1925 (Nematocla: Aphelenchidae). Zool. occurring. The jelly-like mass moved back and Zuhr. 37: 13-19. forth in the intestine with body movements. DAS, V. M. 1960. Studies on the nematode para- Even after continuous feeding for 30 minutes sites of plants in Hyderabad (Andhra Pradesh, there were not more than four or five such jelly India). Z. Parasitenk. 19: 55,3-605. masses in the length of the lumen suggesting FILIPJEV, I. N. 1934. Nematody vreclnye i poli- that the rate of digestion was fast enough to znye v sel'skom khozyaistve. Moscow and Leningrad, Ogiz. 440 pp. keep up with the ingestion, though the nem- GOODEY, J. B. 1958. Paraphelenchus mycelioph- atodes were voracious feeders. thorus n. sp. (Nematoda: Aphelenchidae). The feeding process took 2 to 3 seconds; 1 Nematologica 3: 1-15. second for stylet insertion followed by 1 to 2 MICOLETZKY, H. 1922. Die freilebenclen Erd- seconds of metacorpal pump action. The ac- Nematoden. Arch. Naturgesch. 87 Abt. A tion of the metacorpal bulb allowed the with- (8): 1-320 b; (9): 321-650. drawal of the cell contents and presumably also . 1925. Die freilebenden Siisswasser-und Moornematoden Danemarks. K. danske vi- aided the flow of saliva into the esophageal densk. Selsk. Skr. ser. 8, 10: 57-310. lumen. Thus, food was mixed with the saliva STEINER, G. 1934. Observations on nematocles before it reached the intestine. parasitic in tubers of cinnamon vine (Diasco- Feeding habits of an isolate of Aphelenchus rea batatas}. Proc. Helminthol. Soc. Wash. avenue Bastian, 1865 were studied for com- 1: 15-17.

Four New Species of Gyrodactylus (Tremaloda: Monogenea) from Southeastern U. S.*

THOMAS L. WELLBORN, JR. Bureau of Sport Fisheries and Wildlife, U. S. Department of the Interior, Auburn University, Auburn, Alabama

The host specimens used in this study were rounded lateral anterior projections that extend collected by seine and with HTH (calcium out from bar at approximately a 45° angle hypochlorite). The Gyrodactylus species were (Fig. 3); length 10 (8 to 11) measured to collected using the method described by Putz anterior edge of ventral bar. Shield of ventral and Hoffman (1963), and were treated and bar 17 (15 to 20) long; width at proximal end measured as described by Rogers and Well- 18 (16 to 23); with lateral margins tapering born (1965). to concave distal end. Length of marginal The species described in this paper were hooks 7 to 8 from tip of bilobed base to most collected as part of a survey of fish parasites distant point of curvature (Fig. 4); shaft 30 of National Fish Hatcheries. (28 to 32) long; lamella U-shaped, 11 (10 to All measurements are in microns. Averages 13) long. Cirrus pouch 13 (12 to 16) in are to the nearest micron and ranges are given diameter, on left of ventral midline just pos- in parentheses. The illustrations were drawn terior to pharynx. Cirrus with one large spine with the aid of a camera lucida. and four prominent stylets (Fig. 5). Pharynx 46 (37 to 50) in transverse diameter; bilobed in Gyrodactylus bretinae sp. n. side view with both lobes approximately same (Figs. 1-5) diameter. Each cephalic lobe with a prom- HOST AND LOCALITY: Etheofitoma fitigmaeum inent spine that protrudes a distance of 4 to 5. (Jordan) speckled darter, National Fish Hatch- COMPARISON: Gyrodactylus bretinae resem- ery, Corning, Clay County, Arkansas. bles G. eucaliae Ikezaki and Hoffman, 1957, LOCATION ON HOST: Fins and body. G. heterodactylus Rogers and Wellborn, 1965, SPECIMENS STUDIED: Eleven. G. macrochiri Hoffman and Putz, 1964, and TYPE SPECIMEN: Type and one paratype G. percinae Rogers and Wellborn, 1965, by the USNM Helm. Coll. No. 61626 and 61627. presence of a notch in the dorsal bar. G. per- Paratypes in author's collection. cinae and G. macrochiri have a large C-shapecl DESCRIPTION: Medium-sized gyrodactylid notch in the dorsal bar (fig. 24, Rogers and with fusiform body and smooth thin cuticle; Wellborn, 1965; fig. 1, Hoffman and Putz, greatest length 450 (340 to 490), greatest 1964), whereas G. bretinae has a small simple width 104 (90 to 120). Haptor umbrella- notch in the dorsal bar. It differs from G. shaped, 97 long by 105 wide, with 16 marginal heterodactylus by having marginal hooks that hooks. Anchors moderate, 64 (62 to 67) from are all equal in size. G. eucaliae differs by tip of base to most distant points of curvature having a sickle-shaped marginal hook that (Fig. 1); length of point 27 (25 to 30). Length of dorsal bar 23 (19 to 27), width at lacks a lamella, and a cirrus that possesses eight center 3 to 4; ends of bar slightly constricted small stylets. then expanded into terminal plates 9 to 10 Gyrodactylus campostomae sp. n. long by 5 (4 to 6) wide; center of dorsal bar (Figs. 6-10) with small simple notch on posterior edge (Fig. 2). Length of ventral bar 27 (24 to 29); width HOST AND LOCALITY: Campostoma anomalum (Raf inesque), stoneroller, Chewacla at center 6 (5 to 7); with large, bluntly Creek, Lee County, Alabama. ADDITIONAL LOCALITIES: Elkhorn Creek, * This research was partially supported by the South- eastern Cooperative Fish Parasite and Disease Project and National Fish Hatchery, Frankfort, Franklin carried out in facilities provided by Auburn University, Auburn, Alabama. County, Kentucky.

50 AJ

LOCATION ON HOST: Fins and body. lobe with a spine that protrudes a distance of SPECIMENS STUDIED: Twelve. 2 to 3. TYPE SPECIMEN: Type and one paratype COMPARISONS: Gyrodactylus campostomae USNM Helm. Coll. No. 61628 and 61629. most closely resembles G. rhinichthius Wood Paratypes in author's collection. and Mizelle, 1957, and G. protuberus Rogers DESCRIPTION: Medium-sized gyrodactylid and Wellborn, 1965, by possessing a medial with fusiform body and smooth thin cuticle; knob on the ventral bar. It differs from both greatest length 421 (360 to 480), greatest G. rhinichthius and G. protuberus by having width 72 (55 to 80). Haptor inverted saucer- much larger anchors, larger lateral anterior shaped, 95 (91 to 100) long by 98 (95 to 100) projections, and the shield of the ventral bar wide; with 16 marginal booklets. Anchors and the dorsal bar are of a different shape. large, 78 (74 to 80) from tip of base to point of greatest curvature; length of point 30 (25 Gyrodactylus lineadactylus sp. n. to 33). Tip of anchor base bent toward point '(Figs. 11-15)' of anchor (Fig. 6). Length of dorsal bar 22 HOST AND LOCALITY: Promoxis nigromacu- (18 to 25), width at center 3 to 4; ends of latus (LeSueur), black crappie, Oktibaha dorsal bar expanded into small oval terminal Creek, National Fish Hatchery, Meridian, plates, 5 to 6 long by 4 to 5 wide (Fig. 7); Lauderclale County, Mississippi. dorsal bar of almost uniform width except for LOCATION ON HOST: Body and fins. terminal plates. Length of ventral bar 26 (24 SPECIMENS STUDIED: Seventeen. to 30), width at center 9 (8 to 10); with TYPE SPECIMEN: Type and one paratype massive, bluntly rounded lateral anterior pro- USNM Helm. Coll. No. 61630 and 61631. jections that extend out from ventral bar at Paratypes in author's collection. approximately an 80° to 90° angle; length 15 DESCRIPTION: Medium-sized gyrodactylid (14 to 17) measured to anterior edge of ven- with fusiform body and smooth thin cuticle; tral bar (Fig. 8). Ventral bar with a large greatest length 478 (400 to 520), greatest medial knob 5 to 6 long by 4 to 5 wide. width 86 (80 to 95). Haptor umbrella-shaped, Shield of ventral bar 28 (25 to 30) long; width 90 long by 100 wide. Anchors large, 78 (76 at proximal end 17 (15 to 20); with lateral to 83) from tip of base to most distant point margins tapering slightly outwards to truncated of curvature; length of point 29 (25 to 31) distal end. Ventral shield with six rows of rec- (Fig. 11). Length of dorsal bar 27 (24 to tangular, retractile ridges that are divided into 29), width at center 3 to 4; ends of bar con- two groups of three parallel rows each. Length stricted slightly then expanded into terminal of marginal hook 5 to 6 from tip of slightly plates 10 to 11 long by 5 to 6 wide; center bilobed base to most distant point of curva- of dorsal bar with large simple notch on pos- ture; lamella U-shaped, 14 (13 to 15) long terior edge (Fig. 12). Length of ventral bar and difficult to see except with phase contrast 27 (26 to 29), width at center 8 to 9; with microscope; shaft 39 (36 to 41) long. Cirrus small sharply pointed lateral anterior projec- pouch 18 (15 to 19) in diameter, on left of tions that extend out from bar at approximately ventral midline just posterior to pharynx. Cir- a 45° angle (Fig. 13); length 5 (4 to 6) mea- rus with one large spine and four to five stylets. sured to anterior edge of ventral bar. Shield Pharynx 31 (25 to 35) in. transverse diam- of ventral bar 21 (20 to 24) long; width eter; bilobed in side view with both lobes at proximal end 15 to 16; lateral margins al- approximately same diameter. Each cephalic- most parallel, distal end slightly concave; with

FIGS. 1—20. New species of Gyrodactylus from the Southeastern U. S. Figs. 1-5. Gyrodactylus hre- tinae sp. n. 1, anchor; 2, dorsal bar; 3, ventral bar and shield; 4, marginal hook; 5, cirrus. Figs. 6-10. G. campostomae sp. n. 6, anchor; 7, dorsal bar; 8, ventral bar and shield; 9, marginal hook; 10, cirrus. Figs. 11-15. G. lineadacti/lus sp. n. 11, anchor; 12, dorsal bar; 13, ventral bar and shield; 14, cirrus; 15, marginal hook. Figs. 16-20. G. minytremae sp. n. 16, anchor; 17, dorsal bar; 18, ventral bar and shield; 19, marginal hook; 20, cirrus.

Copyright © 2011, The Helminthological Society of Washington 58 PROCEEDINGS OF THE [Vol.. 34, No. 1 seven to 10 fine striations. Length of marginal edge of ventral bar (Fig. 18); lateral anterior hook 11 (10 to 12) from tip of base to most projections extend out from dorsal bar at ap- distant point of curvature; lamella U-shaped, proximately a 50° to 60° angle. Shield of ven- 21 (19 to 23) long; shaft 31 (29 to 32) long. tral bar 19 (17 to 20) long; width at proximal Marginal hook only slightly recurved and base end 23 (21 to 25); with lateral margins almost almost straight. Lamellae appear to be at- parallel; distal end slightly concave; with seven tached to tip of point of marginal hook; shaft to 10 fine longitudinal striations. Length of of marginal hook passes through arms of marginal hooks 5 to 6 from tip of bilobed base lamella (Fig. 15). Cirrus pouch 13 to 15 in to most distant point of curvature; shaft 26 (25 diameter, on left of ventral midline just pos- to 28) long; lamella U-shaped, 10 to 11 long terior to pharynx. Cirrus with one large spine (Fig. 19). Cirrus pouch 13 (12 to 15) in and four prominent stylets (Fig. 14). Pharynx 37 (35 to 40) in transverse diameter; bilobed diameter, on left of ventral midline just pos- in side view. Spines did not protrude from terior to pharynx. Cirrus with one large spine cephalic lobes in specimens studied. and four to five small stylets. Pharynx 38 (35 COMPARISON: Gyrodactylus lineadactylus to 40) in transverse diameter; bilobed in side may be distinguished from other North Amer- view with both lobes of approximately same ican gyrodactylids by the long lamellae at- diameter. Cephalic lobes with a prominent tached to the tip of the point of the slightly spine that protrudes a distance of 5 to 6. recurved marginal hooks. Also it is unique COMPARISONS: Gyrodactylus minytremae in that the shaft of the marginal hook passes most closely resembles G. eucaliae Ikezaki and through the arms of the lamella. Hoffman, 1957, G. bretinae, and G. lineadactylus on the basis of the dorsal bar possessing Gyrodactylus minytremae sp. n. a simple notch on the posterior edge, but (Figs. 16-20) differs from them in the shape and size of HOST AND LOCALITY: Minytrema melanops the anchors, the size and shape of the ventral (Rafinesque), spotted sucker, Chewacla Creek, bar, and in the shape of the dorsal bar. Lee County, Alabama. Thanks are extended to Messrs. J. H. Hen- LOCATION ON HOST: Fins and body. ning, J. W. Quam, W. F. Ashe, and to the SPECIMENS STUDIED: Nine. many other persons who assisted in collecting TYPE SPECIMEN: Type and paratype USNM the host fish and to Mr. William Smith-Vaniz Helm. Coll. No. 61632 and 61633. Paratypes for confirming the identification of the host in author's collection. fish. DESCRIPTION: Medium-sized gyrodactylid with fusiform body and smooth thin cuticle; SUMMARY greatest length 520 (490 to 570), greatest Four new species of Gyrodactylus are de- width 81 (70 to 90). Haptor umbrella-shaped, scribed from freshwater fishes of the South- 93 (80 to 110) long by 98 (85 to 120) wide. eastern U. S.: G. bretinae from Etheostoma Anchors 63 (61 to 65) from tip of base to stigmaeum (Jordan), G. campostomae from most distant point of curvature; length of Campostoma anomalum (Rafinesque), G. point 25 (23 to 27) (Fig. 16). Length of lineadactylus from Pomoxis nigromaculatiis dorsal bar 25 (23 to 28), width at center 3 (LeSueur), and G. minytremae from Miny- to 4: ends constricted then expand into ter- trema melanops (Rafinesque). All fish hosts minal plates 9 to 10 long by 5 wide; bar with were collected from Alabama, Arkansas, Ken- prominent protrusions on posterior edge just tucky, and Mississippi. medial to constrictions. Dorsal bar with shallow simple notch in center of posterior edge LITERATURE CITED (Fig. 17). Length of ventral bar 30 (28 to HOFFMAN, G. L., AND R. E. PUTZ. 1964. Studies 31), width at center 7 (6 to 8); with large, on Gyrodactylus macrochiri n. sp. (Trema- bluntly rounded lateral anterior projections, tocla: Monogenea) from Lepomis macrochirm. 14 (13 to 16) in length measured to anterior Proc. Helmintriol. Soc. Wash. 31: 76-82.

PUTZ, R. E., AND G. L. HOFFMAN. 1963. Two ROGERS, W. A., AND T. L. WELLBORN. 1965. new Gyrodactylus (Trematocla: Monogenea) Studies on Gyrodactylus (Trematoda: Mono- from cyprinid fishes with a synopsis of those genea) with descriptions of five new species found on North American fishes. J. Parasit. from the Southeastern U. S. J. Parasit. 51: 49: 559-566. 977-982.

Cultivation of the Turkey Coccidium, Eimeria meleagrirnitis Tyzzer, 1929, in Mammalian Kidney Cell Cultures

DAVID J. DOR AN AND JOHN M. VETTERLING Beltsville Parasitological Laboratory, Animal Disease and Parasite Research Division, Agricultural Research Service, U. S. Department of Agriculture, Beltsville, Maryland

Long (1965) obtained complete develop- Bacteria-free oocyst suspensions were ob- ment of the chicken cocciclium, Eimeria tenella, tained by treatment with 5.25% sodium hypo- from sporozoites to oocysts in the chorioal- chlorite (undiluted Clorox) as described by lantoic membrane of embryonated eggs. Pat- Jackson (1964). They were stored in Ringer's ton (1965), however, was the first to obtain solution at 3 to 6 C and were between 2 weeks development of a eoccidium in cell culture. and 3 months old when used. Utilizing monolayer cultures of bovine kidney EXCYSTATION OF SPOROZOITES: SpOl'OCyStS cells (secondary, cell line) and fibroblast-like were released from oocysts by grinding with cells (secondary) derived from 9-day-embryo- a mortar and pestle (Doran and Farr, 1962). nated Japanese quail, he obtained development They were concentrated by centrifugation at of E. tenella sporozoites to mature, first gen- 1,500 g for 5 minutes and then treated for eration schizonts. His was the first report of 15 to 18 minutes at 37 to 41 C with excysta- cultivation of an obligate intracellular intestinal tion solution [5% chicken or turkey bile in protozoan parasite in cell culture. Strout et al. Ringer's solution + 0.25% trypsin' (1-300, (1965) have reported establishment of E. Nutritional Biochemicals Corp.)]. The solu- acervulina sporozoites in monolayer cultures of tion, which had been Seitz filtered, quickly chick embryo kidney, chick embryo fibroblasts, frozen, and stored in 5-ml aliquots at —40 C, mouse fibroblasts, human amnion, and HeLa was adjusted to pH 7.3 to 7.8 and used cells. In a note at the end of their paper, they immediately after thawing. In several pre- added that "segmentation of the trophozoite" liminary tests, between 85 and 95% of the was subsequently observed in some of the available sporozoites excysted during the 15- cultures. to 18-minute interval. This report concerns the cultivation of a CELL CULTURES: The source, origin, and turkey eoccidium, E. meleagrirnitis, in mono- maintenance media of the kidney cell cultures layer cultures of bovine and porcine kidney are listed in Table 1. All cultures were in cells. Leighton tubes containing 10- by 22-mm cover glasses. They were kept at 41 C. Each experi- MATERIALS AND METHODS ment consisted of three to seven cultures for OOCYSTS: Oocysts were obtained from the each time interval. Media also contained phenol droppings of turkeys at the height of infection. red indicator (10 mg/ml), dihydrostreptomycin The method for their recovery and sporulation (100 /xg/ml), penicillin (100 units/ml) and was the same as that previously described nystatin (100 units/ml). The maintenance (Doran and Farr, 1962). Only oocyst suspen- media were changed when necessary to main- sions having more than 85% sporulated oocysts tain the pH in each culture between 6.8 and were used. 7.2.

TABLE 1. Source, origin, and maintenance media of cell cultures.

Exper. Serial passage Maintenance Cell type number Source Origin number medium Bovine ( primary ) 1 F embryon c BME .1 F embryon c BME 3 MA embryon C BME 4 MA embryon c ELAC Bovine ( serial ) 1 MA embryon C 10 BME 2 MA embryon c 17 BME 3 MA embryon c 20 BME Porcine ( primary) 1 MA embryon c BME 2 MA embryon c BME 3 BPL non-embryonic ELAC Porcine (serial) 1 MA non-embryonic 12 BME 2 NADL non-embryonic 17 ELAC 3 BPL non-embryonic 13 ELAC 4 NADL non-embryonic 172 ELAC F — Flow Laboratories, Rockville, Md. MiVl.fAi — IVllL-IUUiUlU^lCiMicrobiologicaUl /ibSUCiiUUaAssociates, AJiriilCSUilBethesda,, IVILIMd.. BPL — Beltsville Parasitological Laboratory, prepared essentially by the method of Younger (1954). NADL = National Animal Disease Laboratory, Ames, Iowa. BME = Basal Medium Eagle (Eagle, 1955) with Earle's (Earle, 1943) balanced salt solution + 2% calf serum. ELAC = Earle's B.S.S. (93%), lactalbumin hydrolysate (5%), and calf serum (2%).

INOCULATION OF CELL CULTURES: Excysted It was found in preliminary work that most sporozoites were concentrated by centrifuga- of the sporozoites that entered cells did so by tion at 1,800 g for 5 minutes, washed once 21/2 to 3 hours. Therefore, after 3 to 3M> hours with the maintenance medium, and then re- at 41 C, the sporozoite-containing suspension suspended in the volume necessary to infect was removed and the cells were washed twice the planned number of cultures. After the with fresh maintenance medium. This re- suspension had been quickly adjusted to pH moved some of the debris (unbroken oocysts, 7.0 to 7.2, it was thoroughly agitated and 1.2 sporocysts, and oocyst and sporocyst hulls) ml was pipetted into each cell culture. Only from the cultures. those cultures that were confluent and did not FIXING AND STAINING CELL CULTURES: At contain cell aggregates were used. 24, 48, 72, 96, and 120 hours after inoculation, Inocula could not be accurately counted and cover glasses were removed from the tubes, calibrated because of differences in release of gently washed in warm phosphate buffered sporocysts, excystation of sporozoites, and es- saline for about 3 seconds, and placed in pecially the survival rate in the maintenance aluminum wire baskets attached to glass slides. media. It was found (unpublished) that the The cultures were then fixed in 10% neutral sporozoites died quite rapidly in the absence phosphate buffered formalin, stained with of cells. Each experiment, however, was Harris' hematoxylin and eosiii as recommended started with sufficient sporulatecl oocysts to by Paul (1960), and mounted as permanent yield theoretically about 15 million sporozoites preparations on glass slides. per tube. Sporozoite-containing media were COUNTING: Counts were made using X645 pipetted quickly into tubes to minimize fluc- magnification. They represent those sporo- tuations in number. zoites and other developmental stages that

FIGS. 1-9. Magnification X 1,840. An eosinophilic globule is indicated by a solid arrow and a residual body by a broken arrow. 1. Two sporozoites within the cytoplasm of a cell. 5 hrs. 2. Two immature schizonts each containing three nuclei. Note that one of them does not have an eosinophilic globule. 72 hrs. 3. Immature schizont containing four nuclei. 48 hrs. 4. Immature schizont with eight nuclei. 48 hrs. 5. A sporozoite and a large, oblong, immature schizont with six nuclei. 72 hrs. 6. Im- mature schizont in which merozoite formation has commenced. The vacuole contains nine merozoites, some of which are still attached by their posterior ends. 72 hrs. 7. Schizont in which merozoite formation has been completed, but the merozoites are still attached to the rather large residual body. 48 hrs. 8. Mature schizont with detached merozoites and a smaller, rounded residual body. Note that the eosinophilic globule still persists. 9. Mature schizont, end view.

FIGS. 10-15. Magnification X 1,840, except Figure 11 (x 2,240) and Figures 14 and 15 (X 430). The eosinophilic globule is indicated by the arrow. 10. Merozoites of the first generation free and within an enlarged cell. 72 hrs. 11. Trophozoite, or "ring stage," of the second generation. A merozoite, partially out of focus, is immediately above the trophozoite. 72 hrs. 12. A large, immature schizont that does not contain an eosinophilic gobule. Note the sporocyst shells in upper right corner. 72 hrs. 13. Another immature schizont in which there is no eosinophilic globule. Merozoite formation has taken place, but they still remain attached by their posterior ends. 72 hrs. 14. Porcine cell culture (serial) showing the presence of sporozoites and the absence of debris in the inocula. 120 hrs. 15. Bovine cell culture (serial) showing the near absence of sporozoites and the presence of large amounts of debris in the inocula. 120 hrs.

TABLE 2. Number of sporozoites and schizonts found in bovine kidney (serial) cell cultures at 48, 72, and 96 hours after inoculation.

48 hours 72 hours 96 hours Schizonts Schizonts Schizonts Exper. Culture Sporo- with without Sporo- with without Sporo- with without zoites e.•8- e•g. zoites e,•g- e•g. zoites e.•K. e,.g. I M I M I M 1 M I M I M 1 1 1,414 48 13 0 0 607 27 16 5 0 380 5 9. 5 0 2 975 41 9 0 0 320 17 27 7 0 172 3 2 1 0 3 1,920 29 27 0 0 870 14 14 9 0 312 9 1 1 0 totals 4,309 118 49 0 0 1,797 58 57 21 0 864 17 4 7 0 2 1 1,112 39 1 0 0 970 19 19 3 0 240 9 0 2 0 2 1,495 18 2 0 0 459 29 10 2 0 467 4 0 0 0 3 2,000 27 4 0 0 1,002 22 12 0 0 325 0 1 4 0 totals 4,607 84 7 0 0 2,431 70 41 5 0 1,032 13 1 6 0 3 1 1,320 27 2 0 0 820 9 9 14 0 175 3 4 2 0 2 1,475 17 9 0 0 740 9 12 15 0 217 1 2 1 0 3 1,505 30 15 0 0 683 7 4 10 0 321 3 2 3 0 totals 4,300 74 26 0 0 2,243 25 25 39 0 713 7 8 6 0 Abbreviations: I = immature; M = mature; e.g. = eosinophilic globule. were within cells on one-half of the cover to 3.8 /A by 1.2 ^,. Trophozoites or "ring stages" glass. The area from which counts were made (Fig. 11) and immature schizonts not having was always every other "row" across the length an eosinophilic ("retractile") globule (Figs. 2, of the cover glass. The presence of a vacuole- 12, and 13) were found at 72 and 96 hours. like intracytoplasmic space around the parasite The trophozoites were round and measured 1.5 was used as criterion of intracellularity. to 2.0 /x, in diameter. The immature schizonts were generally a few microns larger than those RESULTS with a globule. Sporozoites entered cells in all of the pri- The number of schizonts with and without mary and serial cultures of both bovine and an eosinophilic globule found at each of the porcine kidney. They were found in fibroblast- intervals was extremely small (Table 2). At like cells as well as epithelial-like cells. Multi- 48 hours, less than 5% of the parasites in each ple infection of cells with sporozoites was com- culture were schizonts. At 72 hours, it was mon. When infection with four or less oc- 3 to 14%, and at 96 hours, again less than 5%. curred, and all were less than one body length At 120 hours, only a very small number (less from one another, they were usually found side than 100) of sporozoites were present. by side and oriented in. the same direction Between 48 and 96 hours, there was a 69 (Fig. 1). to 89% decrease in the total number of para- Schizogony occurred in only the epithelial- sites found in each of the cultures (Table 2). like cells in serial cultures of bovine kidney. These losses were probably not because of a Although many of the small number of epi- single factor. However, a major cause prob- thelial-like cells present in some of the primary ably was the debris in the inocula. Washing cultures contained sporozoites, schizonts were three times with fresh maintenance medium not observed. was not equally effective in removing the Immature and mature schizonts of the first oocyst and sporocyst hulls from the four types asexual generation (Figs. 2-9) were found at of cultures. Nearly all of the debris was re- 48, 72, and 96 hours. Immature schizonts, in moved from serial porcine cultures (Fig. 14), which merozoite formation had not yet begun but very little was removed from the serial (Figs. 2-5), measured 4 to .16 p. by 4 to 6 p,. bovine cultures (Fig. 15). Sharma and Foster Mature schizonts containing 12 to 28 mero- (1964) found that extracts of E. tenella oocysts zoites that were detached from a large residual were toxic to cells in rabbits. This is probably body measured 13 to 18 p. by 12 to 14 ^. The also true for cells grown in vitro. In the serial merozoites (Figs. 8, 10), each of which con- bovine cultures, cells died quickly after 48 tained a prominent nucleus in the rounded hours and many of the dead cells contained posterior one-half of its body, measured 3.2 degenerate parasites.

DISCUSSION in monolayer cultures of the definitive host cell. An eosinophilic ("ref ractile") globule is It was rather surprising that no develop- considered characteristic of sporozoites and ment was observed in serial porcine cells. The schizonts of only the first asexual generation epithelial-like cells were in excellent condition of E. meleagrimitis (Clarkson, 1959; Tyzzer, and many sporozoites were present after 120 1929). The schizonts that lacked a globule hours. The serial porcine cells, as well as the were found 24 hours later than those with a primary porcine and bovine cells which were globule and may well have been those of the also in good condition and contained sporo- second generation. However, it is not known, zoites, apparently lacked the necessary intrinsic especially in cell culture, whether the eosino- cell stimulus to initiate development. Cessation philic globule regresses in size and disappears of development in the serial bovine cells was before merozoite formation in some schizonts probably because of inadequate cell stimulus. but not in others. The stimulus for further development may Patton (1965) found mature first genera- (1) never have been inherently present in the tion schizonts of E. tenella in bovine kidney cells or (2) diminished or lost because of cell cultures at 4 to 6 days with the greatest inadequate growth conditions imposed on the number at 5 days. In the chicken, 3 days cultures by the debris in the inocula. were required for completion of the first generation (Tyzzer, 1929). The time intervals at SUMMARY which the early schizogonous stages of E. Eimeria meleagrimitis, a parasite of the meleagrimitis were first found in vitro were duodenal epithelium of turkeys, completed one surprisingly similar to those at which similar asexual generation and perhaps part of an- stages are found in the turkey. Clarkson (1959) other in monolayer serial cultures of bovine reported immature and mature first generation kidney cells. Development did not take place schizonts at 24 and 48 hours, respectively. in primary bovine, primary porcine, or serial In vitro, no developmental stages were found porcine cell cultures. at 24 hours, but both immature and mature Although the time intervals at which the schizonts were found at 48 hours. Clarkson early schizogonous stages were first found also found trophozoites of the second genera- in vitro were similar to those at which similar tion at 48 hours and mature schizonts of the stages were found in the turkey, the morphol- same generation at 66 hours. In vitro, tropho- ogy was quite different. Mature first genera- zoites also occurred at 48 hours and immature tion schizonts contained only 18 to 28 mero- schizonts, if actually those of the second generation, were seen at 72 hours. Although the zoites. Immature schizonts, which lacked an time intervals for appearance of developmental eosinophilic ("refractile") globule and might stages in vivo and in vitro were similar, mor- have been second generation, were nearly twice phology of these stages was quite different. the size of those in the host. Clarkson reported 80 to 100 merozoites measuring 4.5 by 1.5 JJL within, a first generation LITERATURE CITED schizont. In vitro, schizonts contained only CLARKSON, M. J. 1959. The life history and 18 to 28 merozoites that measured 2.9 to 3.5 /j, pathogenicity of Eimeria meleagrimitis Tyzzer, by 1.4 p.. Clarkson also found second genera- 1929, in the turkey poult. Parasit. 49: 70-82. DORAN, D. J., AND M. M. FARR. 1962. Excysta- tion mature schizonts measuring only 8 by tion of the poultry coccidium Eimeria acer- 7 fj, and containing 8 to 16 merozoites. In vitro, vulina. J. Protozool. 9: 154-161. the immature schizonts without an eosinophilic EAGLE, W. H. 1955. Nutrition needs of mam- globule were nearly twice as large and many malian cells in tissue culture. Science 122: of them (Fig. 13) already contained more than 501-504. 16 merozoites. Cell culture, especially mono- EARLE, W. R. 1943. Production of malignancy layer, is a highly artificial system. One should in vitro. IV. The mouse fibroblast cultures and changes seen in living cells. J. Am. Can- expect some variation. Even so, it will be inter- cer Inst. 4: 165-212. esting to compare the present morphological JACKSON, A. R. B. 1964. The isolation of viable findings with those of the same species grown coccidial sporozoites. Parasit. 54: 87-93.

LONG, P. L. 1965. Development of Eimeria STROUT, R. G., J. Sous, S. C. SMITH, AND W. R. tenella in avian embryos. Nature 208: 509- SUNLOP. 1965. In vitro cultivation of 510. Eimeria acervulina (Coccidia). Exper. Para- PATTOX, W. H. 1965. Eimeria tenella: Culti- sit. 17: 241-246. vation of the asexual stages in cultured animal TYZZER, E. E. 1929. Coccidiosis in gallinaceous cells. Science 150: 767-769. birds. Am. J. Hyg. 10: 269-384. PAUL, JOHN. 1960. Cell and Tissue Culture. The YOUNGER, J. S. 1954. Monolayer tissue cultures. Williams and Wilkins Co., Baltimore. Preparation and standardization of suspen- SHAHMA, N. N., AND J. W. FOSTER. 1964. Toxic sions of trypsin-dispersed monkey kidney substances in various constituents of Eimeria cells. Proc. Soc. Exp. Biol. and Med. 85: tenella oocysts. Am. J. Vet. Res. 25: 211-216. 202-205.

Intestinal Protozoans and Parasites of the Gelacla Baboon (Theropithecus gelada Riippel, 1835) *

ROBERT E. KUNTZ, BETTY JUNE MYERS, AND THOMAS E. VICE Division of Microbiology and Infectious Diseases, Southwest Foundation for Research and Education, San Antonio, Texas

Even though baboons range throughout the hosts, plus postmortem examination of two greater part of Africa, the monotypic Thero- others. Fecal samples were preserved by the pithecus gelada occurs only in a restricted area MIF (merthiolate-iodine-formalin) vial technic associated with the highlands of Ethiopia. The (Sapero and Lawless, 1953). The incidence gelada, essentially a terrestrial species, occurs for intestinal protozoa and helminths is based in numbers in its habitat and has been known upon a combination of the direct smear from to man for many decades. However, the se- sediments in the vial and the MIFC concentra- clusion of undeveloped areas of Ethiopia and tion technic (Blagg, et al., 1955). A search the exportation of limited numbers of animals for blood parasites was based on an examina- seems to account for the rarity of this primate tion of two thin smears stained in Giemsa. in zoos and zoological gardens. A number of Standard parasitological procedures were em- protozoans and helminth parasites have been ployed for complete postmortem examinations. reported for other species of baboons (Myers All tissues were macerated and washed in sev- and Kuntz, 1965), but there are only a few eral changes of water to allow efficient re- records for the gelada. moval of parasites. Nematodes were cleared The present report, based upon an examina- in phenol to facilitate identifications. tion of a short series of animals in San Antonio Zoo, provides additional information to our RESULTS AND DISCUSSIONS limited parasitological knowledge of T. gelada. The incidence of intestinal protozoans and MATERIALS AND METHODS helminths determined by examination of a single fecal sample from 13 hosts, phis two Thirteen gelada baboons, obtained through autopsy recordings, is presented in Table 1. an animal importer, were examined shortly Oesophagostomum bifurcum, Trichostrongylus after arrival and before introduction into the colubriformis and a pentastome nymph (Lin- San Antonio Zoo. Data and information were guatula serrata) were obtained at the time of obtained by an examination of stools from 13 postmortem examinations. Although the listing * This investigation was supported by research snint HE- is not extensive, it represents the largest sam- 03834-07 National Institutes of Health, U. S. Public Health Service. pling made at one time for the gelada baboons.

TABLE 1. Intestinal Protozoans and Helminths of ings of 2 hosts, revealed 1 parasitic (Entamoeba Theropitheciis gelada. histolytica) and 3 commensal protozoans, as PROTOZOA well as 4 species of nematodes and a larval Endolimax ntinci (Wenyon and O'Connor, 1917) pentastome. Endolimax nana, Entamoeba coli, Brug, 1918 3/13 Entamoeba coli (Grassi, 1879) Casagrandi and lodamoeba biitschlii, Oesophagostomum bifur- Bargagallo, 1895 5/13 cum, Trichostrongyliis cohibriformis, and Lln- Entamoeba histolytica Schaudinn, 1903 1/13 lodamoeba biitschlii (von Prowazek, 1911) guatula serrata are new records for the gelada Dohcll, 1911 1/13 baboon. HELMINTHS Heterodera sp. 1/13 Oesophagostomum bifurciim (Crcplin, 1894) ACKNOWLEDGMENTS Railliet and Henry, 1906* 1/13 Trichostrongyltts colubriformis (Giles, 1892) Ransom,'1911* 2/13 The authors are indebted to Fred Starck, Trichtiris sp. 2/13 Director, San Antonio Zoo, for cooperation in ARTHROPODS making geladas available for examination, to LinKiiatuhi serrata Frolich, 1789 (nymph)* 1/13 V. J. Maida, Parasitology Department, South- * Identification based upon specimens obtained at autopsy. west Foundation for Research and Education, for assistance in processing parasitological ma- Most of the few reports on the parasites of terials, and to Dr. J. Teague Self, University of Theropitheciis have been concerned with hel- Oklahoma for identification of the pentastome. minths, and particularly cestode cysts, removed at postmortem examination of the animals LITERATURE CITED maintained in zoos and zoological gardens BLAGG, W., E. L. SCHOEGEL, N. S. MANSOUR, AND (Myers and Kuntz, 1965). Apparently, only G. I. KHALAL. 1955. A new concentration two protozoans, Entamoeba histolytica and technic for the demonstration of protozoa and Entamoeba chattoni (Salis, 1941) and two helminth eggs in feces. Am. J. Trop. Med. intestinal helminths, Physaloptera turgida and Hyg. 4: 23-28. (Canavan, 1929) and Trick-urns (Fiennes, CANAVAN, W. P. N. 1929. Nematode parasites in the Philadelphia Zoological Garden and 1966) have been previously reported from vicinity. I. Parasitology 21: 63-102. this host. While not unexpected in baboons, FIENNES, R. N. T. W. 1966. Report of the So- the other protozoans and helminths listed in ciety's Pathologist for the year 1964. J. Zool. Table 1 are considered as new host records. 148: 363-380. An examination of anesthetized animals re- KUNTZ, R. E., AND B. J. MYERS. 1966. Parasites vealed that these primates were much cleaner of baboons (Papio doguera Pucheran, 1856) than the many Kenya baboons, Papio doguera captured in Kenya and Tanzania, East Africa. Pucheran, 1856, which we have processed Primates 7: 27-32. previously (Kuntz and Myers, in press), and KUNTZ, R. E., AND B. J. MYERS. Microbiology there were no ectoparasites. No parasites were parameters of the baboon (Papio). (In detected in vaginal samples or in peripheral Vagtborg, H., ed. The baboon in medical blood smears. In contrast with the conditions research. Vol. II. Proceedings of Second generally present in P. doguera (Kuntz and International Symposium on the baboon and Myers, unpublished data), oral examinations its use as an experimental animal. University of the geladas revealed unusually clean mouths, of Texas Press, Austin ) (in press ). MYERS, B. J., AND R. E. KUNTZ. 1965. A check- with the absence of Trichomonas and Enta- list of parasites reported for the baboon. Pri- moeba. The latter commensals frequently are mates 6: 137-194. associated with the accumulation of tartar, SALIS, H. 1941. Studies on the morphology of organic detritus and lesions in captive P. the E. histolytica-like amoebae found in mon- doguera. keys. J. Parasitol. 27: 327-341. SAPERO, J. J., AND D. J. LAWLESS. 1953. The SUMMARY MIF stain-preservation technic for the identifi- An examination of fecal samples from 13 cation of intestinal protozoa. Am. J. Trop. Theropitheciis gelada, plus postmortem find- Med. and Hyg. 2: 613-619.

Monordotaenia nom. nov. for the Badger Taeniid Cestodes with One Row of Hooks

JOHN W. LITTLE Department of Biology, Texas A & M University, College Station, Texas

Rausch (1947) pointed out the uniqueness Upon reviewing Fossor angertrudae pre- of a single row of hooks in the genus Taenia served on three slides marked USNM No. 9053, in his reclescription of Taenia taxidiensis the paratype from the Wyoming Agricultural Skinker, 1935, from the badger, Taxidea taxus Experimental Station and the paratype of taxus. This redescription was in order since Taenia. taxidiensis on six slides marked 32840, the original paper on T. taxidiensis apparently I have concluded that F. angertrudae is synon- was based on one complete specimen and sev- ymous with T. taxidiensis. eral fragments. Skinker (1935), not knowing The genus Fossor, containing the single that the worm normally contained only a single member F. angertrudae, was created to sep- row of hooks, erroneously assumed that the arate the species from Taenia, a genus in which "other row" had been lost and placed the all members possess a double crown of hooks. eestocle in the genus Taenia. It has already been pointed out that Skinker Later, Honess (1937) found four specimens did not realize she was working with material of taeniid containing a single row of hooks, that contained only one row of hooks and also from the badger, which he described and therefore she naturally placed the worm in named Fossor angertrudae. Wardle and Mc- the genus Taenia. However, since Honess' Leod (1952) referred to F. angertrudae as a generic name, Fossor, is a junior homonym "small form" since its length appeared as 39 of Fossor Liechtenstein, 1844, which is pres- to 46 mm in the original description. The ently a synonym of the rodent genus Geonjchm measurements should have probably read 39 Illiger 1833, Monordotaenia nom. nov. is pro- to 46 cm since the holotype, USNM No. 9053 posed here for the badger cestode containing is 382 mm in length. However, these latter a single row of hooks. The correct name authors suggested that F. angertrudae be re- becomes Monordotaenia taxidiensis, Skinker, garded as "sub juclice" until further records 1935, new combination. were established. DISCUSSION Skinker's holotype apparently has been lost and due to a mixup was not assigned 39803, The major reason why this badger worm has the number in her publication. It now becomes been confused is that the paucity of Skinker's necessary to designate the paratype slide TABLE 1. Comparison of Taenia taxidiensis and USNM No. 32840 as the lectotype, although Fossor angertrudae. this specimen is not complete. The paratype Worm T. taxidiensis F. angertrudae USNM No. 32840 #1, marked hooks, is defec- Source Rausch, 1947 No. USNM 9053 tive as no hooks are present. Therefore, Table Strohila (mm) 480.0 382.0 Width (mm) 3.0 2.6 1 is based on Rausch's redescription of T. Segments gravid taxidiensis and measurements of the holotype ( length mm ) 8.0-8.5 7.3 Scolex ( fj. ) 570.0 780.0 of F. angertrudae, Honess, 1937, USNM No. Suckers ( /i ) 156.0 200.0 No. of hooks 20-27 20 9053, by the present writer. The measure- Size of hooks (fa) 90.0 89.0 Hook guards ( ft ) 46.0 43.0 ments of the two worms, except the scoleces Ventral canals ( /i ) 142.0 140.0 and suckers, are strikingly similar. The excep- Dorsal canals (/u) 20.0 18.0 Cirrus length ( /A) 240.0-330.0 270.0 tions are no doubt clue to techniques in prep- Cirrus width ( /i ) 100.0 110.0 No. of testes 200-300 200-300 aration as the scolex and suckers in Skinker's Size of testes ( p. ) 70.0-90.0 60.0-80.0 Host Taxidea taxtts Taxidea taxus paratype USNM No. 32840 measured 740 and (Schreber) (Schreber) 200 mm, respectively. Locality Wisconsin Wyoming

Copyright © 2011, The Helminthological Society of Washington [VOL. 34, No. 1 material prevented her from making a correct kindly provided me with the material employed assessment in the original publication. Wardle in this study. and McLeod mentioned that Skinker's badger cestode contained one row of hooks, but it LITERATURE CITED was nevertheless placed in Taenia, i.e., with HONESS, R. F. 1937. Un nouveau cestode: Fos- worms containing two circles of hooks. It is sor angertrudae n. g., n. sp. du blaireau surprising that Rausch, in his redescription, did d'Amerique Taxidea taxus taxus (Schreber, not remove this badger worm from the genus 1778). Ann. Parasitol. Hum. Comp. 15: 363- Taenia. 366. It would seem unlikely that the other taeniid LINSTOW, O. F. B. VON. 1905. Helminthen der with a single row of hooks, Taenia rnono- russischen Polar-Expedition 1900-1903. Mem. stephanos Linstow 1905, reported in the lynx Acad. Imp. Sci. St. Petersburg 18: 1-16. from Russia, belongs to the genus Taenia. RAUSCH, R. 1947. A redescription of Taenia However, until more information is accessible, taxidiensis Skinker, 1935. Proc. Helm. Soc. Washington 14: 73-75. this worm must be regarded as a species SKINKER, M. S. 1935. Two new species of tape- inqiiirenda. worms from carnivores and a redescription of ACKNOWLEDGMENTS Taenia laticollis Rudolphi, 1819. Proc. U. S. Nat. Museum 83: 211-220. I wish to thank Dr. W. W. Becklund of the WARDLE, R. A., AND J. A. McLEOD. 1952. The Beltsville Parasitological Laboratory and Dr. zoology of tapeworms, University of Minne- R. F. Honess, University of Wyoming, who sota Press, Minneapolis, USA, 780 pp.

Feeding of Xiphinema index and X. diver sicaudutum

]. M. FISHER* AND D. J. RASKI**

Information on feeding and feeding sites of men of X. diversicaiidatiim could penetrate as Xiphinema spp., vectors of plant viruses, would far as the stele of small roots (< 0.25 mm be useful in studying acquisition and trans- diam.), but further observations are necessary mission of the viruses by the various stages of to substantiate that Xiphinema spp. are reg- the nematodes. Indirect observations (Pitcher ularly capable of such deep feeding in roots. and Posnette, 1963) and pathological symptoms (Davis and Jenkins, 1960; Schindler and METHODS Braun, 1957; Schindler, 1957; Raski and Rade- Feeding of X. index was observed on seed- wald, 1958; Radewald, 1962) indicate some of ling grapes, Vttis vinifera L., var. Mission, the feeding sites of Xiphinema index and growing in 0.75% distilled water-agar culture Xiphinema diversicaiidatiim on certain hosts. in petri dishes. The initial agar medium pre- Feeding of X. index on roots of grape growing pared with copper-distilled water was lethal to on 2% agar was observed directly (Radewald, the nematodes, killing them in six hours. The 1962; Radewald and Raski, 1962) but com- effect -was due presumably to copper ions (van plete and detailed description of the activities Gundy and Thomason, 1962) and glass-distilled are lacking. Pitcher and Posnette (1963) water was used thereafter without difficulty. showed that the onchiostyle of a single speci- Several agar concentrations were tried, with little effect on the nematodes, but because of * Department of Plant Pathology, Waite Agricultural Re- search Institute, Adelaide, South Australia. the high evaporation rate at Davis, it was found ** Department of Nematology, University of California, Davis, USA. that a low agar concentration (0.75% Difco

Copyright © 2011, The Helminthological Society of Washington JANUARY, 1967] HELMINTHOLOGICAL SOCIETY 69 agar) allowed the culture medium to be kept longer in a condition suitable for the growth of the nematodes and the seedlings. No attempt was made to work under aseptic conditions. To reduce the problem of microorganisms, no nutrients were added to the agar medium. Germinating grape seedlings can grow for three to four weeks on the reserve food of the cotyledons. Gall formation on roots, egg- laying, embryological development, moulting and feeding all occurred in one or other of the cultures. All cultures were kept on the laboratory bench at 20 to 22 C.

OBSERVATIONS Fig. 1. Young larval stage of X. index feeding The total time for embryological develop- in the piliferous region of a grape root. ment to hatching was 6 to 8 clays. No moult was observed in the egg which agrees with the observations of Radewald (1962) who microscope light or movement of the petri dish found the first-stage larvae moulted 24-48 disturbed some of the feeding nematodes. hours after emergence. Mature larvae within After a short time, even in the light, most of the egg had a second onchiostyle inserted the nematodes resumed feeding, either at the within the extension of the first stylet in such same or a different site. The older larval a way that the tip of the second onchiostyle stages and adult females (no males were pres- almost reached to the base of the first odonto- ent in the population used) always fed at the style. On hatching the presence of the second root tip, mainly slightly in advance of, or onchiostyle did not prevent feeding. slightly behind, the meristematic cells (Fig. 2). Precise measurements were not taken which The onchiostyle penetrated to the meristem would permit exact identification of hatched either through the root cap cells or through larvae as to first- or second-stage. Conse- that part of the root epidermis just behind the quently these are referred to as newly-hatched root cap, where the diameter of a grape root larvae. These newly-hatched larvae fed well narrows considerably. Pauses in the progress back from the root tip on the outer cortical of the onchiostyle while penetrating outer cor- cells of the piliferous region (Fig. 1). The tical cells, accompanied by muscular contrac- onchiostyles of these larvae were not long tions of the oesophageal bulb appeared to indi- enough to penetrate deeper into the root and cate injection or withdrawal of material to or in this region no symptoms were produced. from the root cells. No material was ever Shortly before moulting they migrated to the observed to pass along the lumen of the oesoph- root tip where they fed for a short time, and agus in either direction but contractions of the small galls were produced. After this short oesophageal bulb frequently were seen. Often feeding period these larvae migrated into the in penetration, the stylet would cease moving medium in advance of the root tip and moulted. and muscular contraction of the posterior part The same two areas of the root were fed 011 of the oesophagus would take place for a vari- by the next larval stage but these larvae showed able length of time up to 2 minutes in each greater ambivalence regarding feeding site successive cell as the onchiostyle was pushed preference, migrating back and forth from deeper into the rootlet. The onchiostyle was piliferous region to root tip. Again no symp- not limited to straight penetration but had toms were produced by feeding in the pilif- enough lateral movement to allow penetration erous region, but galls were formed as a result of adjacent cells in the same optical plane. of feeding at the root tip. Bending of the onchiostyle as shown by Pitcher The time spent at one feeding site was quite and Posnette (1963), while not common, was variable and in some cases switching on of the seen on a number of occasions. Penetration of

2QQM,

•

Fig. 3. Demarcation of the epidermal cell walls of a grape root as a result of nematode feeding.

tip, elongation of the root was prevented, resulting in a swollen root tip with a reduced number of root cap cells and a truncated appearance. When feeding took place on only one side of the root tip, galling was restricted to one side. The shortest time for visible gall formation was 24 hours from the time of inoc- Fig. 2. X. index feeding in the meristematic ulation with the nematodes. Large galls, which region of a grape root tip. are common in pot cultures and in the field, did not develop in petri dish cultures. In all two adjacent cells involved slight withdrawal cultures, nematodes ceased feeding as soon as of the onchiostyle, lateral movement and fur- the roots stopped growing. ther penetration. The foregoing descriptions refer to the feed- The onchiostyle often penetrated very deep- ing of X. index and the work was done at ly but in no case were stylet extensions exserted. Davis, California. Similar observations have Exsertion of the stylet reached its maximum been made recently with X. diversicaudatum in when its base was at the level of the base of Adelaide. Attempts to feed the nematodes on the lips; thus the length of onchiostyle pene- some herbaceous hosts of viruses (cucumber, trating the root depended on the age of the petunia and white clover) failed, but the nema- nematode. The maximum depth of penetration todes did feed and produce galls on rose roots. also depended on the diameter of the root. X. diversicaudatum was invariably more sensi- For feeding to take place, it was not necessary tive to disturbance than X. index, but feeding for the onchiostyle to be fully exserted. sometimes recommenced a short time after ex- An effect on the walls of epidermal cells of posure to a new set of conditions. the root, following passage of the onchiostyle Although feeding at each site was quite was noticed. Initial penetration by the onchio- variable, one example is given in detail. Once style appeared to be mainly intercellular. The the lips of the nematode made suitable contact epidermal cell walls became markedly outlined with the root the onchiostyle penetrated the (Fig. 3) indicating some change in structure, epidermal cell almost immediately. This was perhaps even slight separation of the cells. This followed by intermittent contractions of the effect was observed only at root tips. basal region of the oesophagus of approxi- Some large galls were formed on the root mately 2-second duration; between contractions tips (Fig. 4). Where a number of nematodes were periods of inactivity. This lasted for 5 had fed on different sides of the same root minutes at the end of which time continuous

Copyright © 2011, The Helminthological Society of Washington JANUARY, 1967] HELMINTHOLOGICAL SOCIETY 71 contractions of the bulb lasted for 2 minutes. When the bulb stopped its action, the onchiostyle was pushed deeper into the second layer of cells. The intermittent contractions of the oesophageal bulb then lasted for approximately 1 minute and this was immediately followed by successive but gradually lengthening periods of pulsation of the bulb. The onchiostyle remained in this cell for 3 minutes and then penetrated the next layer of cells. This procedure was repeated as the onchiostyle was pushed deeper into the root. When contact with the root was broken, the lips often lost 200// contact with the root surface before the onchiostyle was fully withdrawn, so that the tip Fig. 4. Galling of grape root tips in petri dish of the onchiostyle was washed by the fluid of culture. the medium.

DISCUSSION or that the host—parasite relation on these plants may be different. Although many observations were possible, the disadvantages of the limited period of SUMMARY growth of the host were considerable. In following the life cycle of X. index it was Feeding of Xiphinema index and X. diversi- necessary to remove the nematodes to a new caudatum on roots of germinating seedlings seedling every three to four weeks. Once the in petri dish cultures is described. Young lar- onchiostyle had penetrated deeply, grape roots vae of X. index fed in the piliferous region as were too thick for viewing the actual cell in well as at the root tip but galls were produced which feeding was taking place. Perhaps an- only at the tips. The presence of a second other host would be more suitable. onchiostyle within the stylet extension of these The problem of relating the habits of nema- larvae did not prevent feeding. Females fed todes in culture to those in soil still exists. only at the root tip and galls were evident Thus the large galls which are observed in pot within 24 hrs after inoculation. Maximum cultures and in the field were not seen in cul- observed exsertion of the stylet was when the ture. This could have been due to a more base of the onchiostyle was level with the transitory type of feeding in culture, smaller base of the lips and thus maximum penetration numbers of nematodes, or the different condi- of the root depended on the length of the ditions and nutrient status for growth of roots. stylet. X. dwersicaudatum was more sensitive Symptoms were produced when young lar- to disturbance while feeding than X. index. vae fed at root-tips but not when the same larvae fed in the piliferous region of the root. ACKNOWLEDGMENTS This suggests that immature, undifferentiated We thank Mr. J. W. Meagher of the Vic- root cells are necessary for symptom produc- torian Department of Agriculture for supply- tion. Feeding in these different sites may also ing X. dwersicaudatum. This research was affect the efficiency of different stages to ac- supported in part by Research Grant No. quire and transmit viruses. During experiments 103746.04 of U.S. PHS, National Institutes on transmission of viruses to herbaceous hosts of Health. symptoms usually are not produced by the nematodes. While this may be due to feeding LITERATURE CITED only in the piliferous regions of these hosts the DAVIS, R. A., AND W. R. JENKINS. 1960. Nema- failure to observe feeding at any site on her- todes associated with roses and the root injury baceous hosts in these experiments suggests caused by Meloidogijne hapla Chitwoocl, 1959, that there may be a different feeding process, Xiphinema diversicaudatum (Micoletzky, 1927)

Thome, 1939 and Hellcotylenchus nanus Stei- RASKI, D. J., AND J. D. RADEWALD. 1958. Repro- ner, 1945. Bull. Md. Agric. Exp. St. A. 106, duction and symptomatology of certain ecto- 16 pp. parasitic nematodes on roots of Thompson PITCHER, R. S., AND A. F. POSNETTE. 1963. Vas- seedless grape. Pi. Dis. Reptr. 42: 941-943. cular feeding by Xiphinema diversicaudatum SCHINDLER, A. F. 1957. Parasitism and patho- (Micol.). Nematologica 9: 301-302. genicity of Xiphinema diversicaudatum, an RADEWALD, J. D. 1962. The biology of Xiphi- eetoparasitic nematode. Nematologica 2: 25- nema index and the pathological effect of the 31. species on grape. Ph.D. Thesis, University of SCHINDLEH, A. F., AND A. J. BRAUN. 1957. California, Davis, California. Pathogenicity of an eetoparasitic nematode, RADEWALD, J. D., AND D. J. RASKI. 1962. A Xiphinema diversicaudatum, on strawberries. study of the life cycle of Xiphinema index. Nematologica 2: 91-93. Phytopathology 52: 748 (abstr.). VAN GUNDY, S. D., AND I. J. THOMASON. 1962. RADEWALD, J. D., AND D. J. RASKI. 1962. Studies Factors influencing storage life of Hemi- on the host range and pathogenicity of Xiphi- cycliophora arenaria, Pratylenchus scribneri nema index. Phytopathology 52: 748-749 and Trichodorus christiei. Phytopathology 52: (abstr.). 366-367.

On the Classification of the Insect Parasitic Nematodes of the Sphaerulariidae Luhhock, 1861 (Tylenchoidea: Nematoda)

W. R. NlCKLE1

One of the first obligate tylenchid parasites 1963b). Fuchs, Wachek, and Riihm each pub- of insects was described by Dufour in 1837 lished comprehensive works, and did much to as Sphaerularia bombi from the bumblebees, clarify the systematic^ of this family. Bovien's Bom-bus terrestris and Bombus hortorus. In publications on insect nematodes were out- 1861, Lubbock proposed the family Sphaeru- standing and are classics today. lariaceae to contain the genus Sphaerularia. At present, 4 subfamilies, 21 genera, and Other significant early contributions to the 116 species are included in the Sphaerulariidae. taxonomy of these forms include: Leuckart Generally the classification is strong at the (1884), von Linstow (1890), zur Strassen generic level, but at the specific level it is (1892), Cobb (1920, 1921), Micoletzky (1922), rather weak because detailed descriptions of T. Goodey (1930), Filipjev (1934), Thorne free-living males and females are lacking. A (1935), Fuchs (1915, 1929, 1933, 1938), Bovien (1937), Currie (1937), Chitwood and tylenchid species can generally parasitize hosts Chitwood (1937), Christie (1938) and Schnei- in one genus, and unlike the mermithids, der (1939). More recent contributions in- adults are found in adult hosts. Therefore, clude: T. Goodey (1953), Wachek (1955), knowledge of the host aids in identification. Riihm (1956), Massey (1956, 1957, 1958, In several of the nematode genera the dorsal 1960, 1962), J. B. Goodey (1956, 1963), esophageal gland orifice is located at least one Khan (1957a, 1957b, 1960), Skarbilovich (1947, stylet length behind the base of the stylet or 1959), Welch (1959), and Nickle (1963a, could not be found. The subventral esophageal glands may empty into the lumen of the esophagus a short distance behind the dorsal gland 1 Research Nematologist, Crops Research Division, Agri- cultural Research Service, U. S. Department of Agriculture, orifice. This led Wachek (1955) to place Beltsville, Maryland. Some studies were made while the author was Research Assistant, University of California, Sphaerularia, Tripius, and Scatonema in the Davis, California, and also Research Officer, Canada De- partment of Agriculture, Belleville, Ontario. Aphelenchoidea, though they lack the typical

aphelenchoid median bulb and in all other Because of the brevity of the original de- respects appear to be tylenchoid. scription, the family diagnosis is emended. Recently, I observed the dorsal esophageal Brief diagnoses of the sub-families and genera gland orifice in the fourth stage female larva are given and a new genus is established. of Sphaerularia bombi and found that it was Synonymies are presented along with my com- located behind the stylet in the normal tylen- ments on the status of controversial groups. choid position. Further, the presence of tylen- The characters that typify the family in- choid spicules and a gubernaculum in these clude: absence of a valved median bulb; forms also, supports their tylenchoid status. unique ovary of the free-living female; en- For these reasons I question the aphelenchoid largement of female nematode in the body status of these genera, and I now consider cavity of host, resulting in the nematode be- some of the genera placed in the Aphelen- coming a reproductive sac (heteromorphism), choidea as belonging to the Tylenchoidea. or female nematode with a prolapsed swollen The problem inherent in making this change uterus; obligate insect or mite parasitism. is that Sphaerulariaceae Lubbock, 1861 is an FAMILY : older family name than Allantonematidae Sphaerulariidae Lubbock, 1861. (Pereira, 1931) Chitwood and Chitwood, 1937. Syn. Sphaerulariaceae Lubbock, 1861. In my opinion the insect parasitic Tylenchoidea DIAGNOSIS (Emended): Tylenchoidea. Usu- merit only family rank along with the plant ally with three distinct forms. Two free-living, parasitic families; Tylenchiclae, Heteroderidae, slender, 0.5-1.0 mm long, often found in hab- Criconematidae, Neotylenchidae, Hoplolaimi- itat of young host and one adult parasitic form dae, and Tylenchulidae. Future work in the found in haemocoel of insect or mite. (1) group may necessitate the creation of a super- FL$*: Stylet or pseudostylet rarely absent. family but at present, all insect-parasitic Tylen- Esophagus without median valvular bulb; am- choidea should be placed in the family Sphaer- pullae near gland orifices often strongly de- ulariidae Lubbock, 1861; the Allantonematidae veloped; dorsal gland orifice not always found (Pereira, 1931) Chitwood and Chitwood, 1937 immediately behind spear, may be midway should be given the rank of subfamily. between spear and subventral gland orifices; Entaphelenchus Wachek, 1955 and Pera- esophageal glands long, overlap intestine. Go- phelenchus Wachek, 1955 are endoparasitic nad diagnostic; prodelphic; ovary small, finger- tylenchids, having the typical aphelenchoid like with few oocytes; oviduct short; uterus median bulb and spicules which are shaped prominent, temporarily packed with individual like rose thorns. The gubernaculum is lacking, sperm after copulation. Becomes infective and in the writer's opinion, only these two stage. Eggs absent. (2)FL3: Usually slightly genera are typical of the Aphelenchoidea. longer than female, not infective. Stylet may During the last 8 years, I have studied rep- be absent. Esophagus weakly developed. Spic- resentative specimens from 15 of the 22 genera ules usually tylenchoid, may be elaborate. belonging to the families Sphaerulariidae and Gubernaculum usually present. Caudal alae Allantonematidae before making the revisions often present and peloderan. (3) AP?: Ob- contained herein. It is hoped that this com- ligate parasite in haemocoel of insects or mites. pilation will stimulate work in this interesting Large swollen reproductive sac, produced by but neglected group. expansion of free-living female; or, with uterus Economically, insect parasitic nematodes of prolapsed, swollen. Stylet present or rarely the Sphaerulariidae annually reduce insect absent, may be retracted. Esophagus and in- populations by untold millions, and are indeed testine usually degenerate at expense of repro- important self-perpetuating biological control ductive system which often fills 80% of body. agents. However, little effort has been made Oviparous or ovoviviparous; ovary usually in manipulating, or exploiting these parasites flexed one or more times; rachis common. for biological control, but the time is ripe for * FL$ : Free-living female. FLcT: Free-living male. this type of endeavor. AP9= Body cavity adult parasitic female.

PROPOSED CLASSIFICATION OF THE SPHAERULARIIDAE Sphaerulariidae Lubbock, 1861. Syn. Sphaerulariaceae Lubbock, 1861. Sphaerulariinae (Lubbock, 1861) Pereira, 1931. *Sphaerularia Dufour, 1837. *Sphaerulariopsis Wachek, 1955. *Tripius Chitwood, 1935. Syn. Asconema Leuckart, 1886. Atractonema (Leuckart, 1886) Leuckart, 1887. Proatractonema Bovien, 1944. Allantonematinae Pereira, 1931. (Emended Chitwood, 1935). Syn. Allantoneminae Pereira, 1931. Allantonematidae (Pereira, 1931) Chitwood and Chitwood, 1937. Contortylenchidae Ruhm, 1956. Allantonema Leuckart, 1884. Syn. Tylenchomorphiifi Fuchs, 1915. * Aphelenchulus Cobb, 1920. *Bovienema Nickle, 1963. *Bradynema zur Strassen, 1892. *Chondronema Christie and Chitwood, 1931. *Contortylenchus Ruhm, 1956. *Heterotijlenchus Bovien, 1937. *Howardula Cobb, 1921. Syn. Tylenchinema T. Goocley, 1930. *Prothallonema Christie, 1938. Metaparasitylenchns Wachek, 1955. n. grad. *Neoparasitylenchus n. gen. Parasitylenchoid.es Wachek, 1955. *Parasitylenchus Micoletzky, 1922. Syn. Polymorphotylenchus Riihm, 1956. Proparasitijlenchus Wachek, 1955. n. grad. Protylenchus Wachek, 1955. *Sulphuretylenchiis Riihm, 1956. n. grad. Scatonema Bovien, 1932. Dotylaphus Andrassy, 1958. (genus inquirenda). Fergusobiinae J. B. Goodey, 1963. Syn. Fergusobiidae Siddiqi and J. B. Goodey, 1963. *Fergusobia Currie, 1937. Syn. Anguillulina (Fergusobia) Currie, 1937. lotonchiinae T. Goodey, 1953. Syn. lotonchiidae (T. Goodey, 1953) Skarbilovich, 1959. *Iotonchium Cobb, 1920.

* Specimens of these genera were studied for this review.

SUBFAMILY: OTHER SPECIES: Sphaerulariinae (Lubbock, 1861) Pereira, S.dendroctoni (Massey, 1956) Nickle, 1963. 1931. Syn. Sphaerularia dendroctoni Massey, DIAGNOSIS (Emended): Sphaerulariidae. 1956. FL 9 : Stylet present, with or without knobs. S. hastatus (Khan, 1957) Nickle, 1963. Without eggs in uterus. FLc$: With tylen- Syn. Sphaerularia hastata Khan, 1957. choid anterior end. Stylet present, with or Stictylus hastatus (Khan, 1957) without knobs. Caudal alae present or absent. Khan, 1960. Spicules and gubernaculum tylenchoid. AP9: S. pini (Fuchs, 1929) Nickle, 1963. Uterus everted partially or completely. Syn. Tylenchus sulphureus pini Fuchs, Genus Sphaerularia Dufour, 1837 1929. (Figs. 2Z, 3V, 5A) Allantonema sulphureus pini (Fuchs, 1929) Filipjev, 1934. FL 9 : Stylet present, without basal knobs. Parasitylenchus sulphureus f. pini Dorsal gland orifice opens just behind stylet. (Fuchs, 1929) Schneider, 1939. FL $ : Stylet present, not prominent, without Allantonema pini (Fuchs, 1929) knobs. Caudal alae absent. Spicules and Wachek, 1955. gubernaculum tylenehoid. AP9 : Uterus com- Stictylus pini (Fuchs, 1929) Ruhm, pletely everted, greatly enlarged, larger and 1956. not as smooth as Sphaerulariopsis, becomes S. piceae (Fuchs, 1929) n. comb. 15,000-20,000 times the volume of original Syn. Tylenchus sulphureus piceae Fuchs, female body which remains as a minute ap- 1929. pendage to uterus, though often wrinkled and Allantonema sulphureus piceae deformed. Oviparous. (Fuchs, 1929) Filipjev, 1934. HOST INSECTS: Parasitylenchus sulphureus f. piceae Hymenoptera; Bombus, Vespa, "Psithyrus. (Fuchs, 1929) Schneider, 1939. TYPE SPECIES: Stictylus sulphureus piceae (Fuchs, Sphaerularia bombi Dufour, 1837. 1929) Ruhm, 1956. Syn. Tylenchus bombi (Dufour, 1837) Cobb, 1890. S. piniphili (Fuchs, 1929) Nickle, 1963. Syn. Tylenchus sulphureus piniphili Genus: Sphaerulariopsis Wachek, 1955 Fuchs, 1929. (Figs. 2W, 3W, 5A) Allantonema sulphureus piniphili (Fuchs, 1929) Filipjev, 1934. FL 9 : Stylet present, well-developed, with Parasitylenchus sulphureus f. pini- basal knobs which may be irregular in size. phili (Fuchs, 1929) Schneider, Dorsal gland orifice opens anteriorly behind 1939. stylet. FL<4: Stylet present, with knobs. Cau- Stictylus piniphili (Fuchs, 1929) dal alae peloderan. Spicules and gubernaculum Ruhm, 1956. tylenehoid. AP 9 : Uterus completely everted, S. ungiilacaudus (Khan, 1957) Nickle, 1963. greatly enlarged, smoother and not as large as Sphaerularia, original female body wrinkled Syn. Sphaerularia ungulacauda Khan, 1957. and deformed, remains as an appendage to Stictylus ungulacaudus (Khan, 1957) uterus. Oviparous. Khan, 1960. HOST INSECTS: Coleoptera; Ernobius, Pi-ssodes, Dendrocto- This genus may eventually be placed under nus. Hymenoptera; Coeloides. Sphaerularia, but is easily distinguished at this TYPE SPECIES: time by the smaller size of the prolapsed Sphaerulariopsis stammeri Wachek, 1955. uterus, the knobbed stylet, the presence of the Syn. Stictylus stammeri (Wachek, 1955) caudal alae. It is normally parasitic in bark Ruhm, 1956. beetles.

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Genus: Tripins Chitwood, 1935 Genus: Allantonema Leuckart, 1884 Syn. Asconema Leuckart, Syn. Tylenchomorphus 1886. Fuchs, 1915. Atractonema (Leuck- (Figs. 1A, IF, 3A, 3B, 4A) art, 1886) Leuckart, FL 9 : Lips set off. Stylet well-developed. 1887. Ovary with few cells. Excretory pore posterior Proatractonema Bo- to nerve ring. FL $ : Lips weakly set off. Sty- vien, 1944. let present. Caudal alae narrow, peloderan. (Figs. 2BB, 2GC, 3Q, 3Y, 5A) Gubernaculum small. AP 9 : Bean-shaped, FL9: Stylet well-developed, with knobs. only 2-3 times as long as wide. Stylet retracted Two long esophageal glands. Ovary few-celled; into body. Oviparous or ovoviviparous. Vulva walls of uterus with large cells; vulva posterior. terminal. FL $ : Stylet faintly developed, without knobs. HOST INSECTS : Caudal alae absent. Spicules and gubemac- Coleoptera; Hylobius, Hijlastes, Philonthus, ulum small, tylenchoid. AP 9 : Fusiform. Ochthebius, Geotrupes. Diptera; Musca. Length up to 0.9 mm. Uterus partially pro- TYPE SPECIES: lapsed. Mouth cone and tail papilla present. Allantonema mirabile Leuckart, 1884. Stylet present, not retracted. Oviparous. Syn. Tylenchomorphus mirabills (Leuck- HOST INSECTS: art, 1884) Fuchs, 1915. Diptera; Cecidomyia or Dasine-ura, Bradysia, OTHER SPECIES: Sciara. A. matthesi Wachek, 1955. TYPE SPECIES: A. morosa (Fuchs, 1929) Filipjev, 1934. Tripius gibbosus (Leuckart, 1886) Chitwood, Syn. Tylenchus morosus Fuchs, 1929. 1935. Parasitylenchus morosus (Fuchs, Syn. Asconema gibbosiim Leuckart, 1929) Filipjev and Sch. Stek., 1886. 1941. Atractonema gibbosum (Leuckart, A. muscae Roy and Mukherjee, 1937. 1886) Leuckart, 1887. A. philonthi Wachek, 1955. OTHER SPECIES: A. silvaticum von Linstow, 1892. Tripius sciara (Bovien, 1944) Wachek, 1955. A. stricklandi Roy and Mukherjee, 1937. Syn. Proatractonema sciara Bovien, 1944. It is possible that A. muscae and A. strick- Poinar (1965) has shown that the morpho- landi are actually the same species, i.e., two logical basis for the genus Proatractonema may female stages of a Heterotylenchus. be just a stage in the development of a Tripius and he considers Proatractonema as a synonym Genus: Aphelenchulus Cobb, 1920 of Tripius. (Figs. 1C, 3C, 4A) SUBFAMILY : FL9: Not found. FLS: Stylet present. Allantonematinae Pereira, 1931. (Emended Length 1 mm. Caudal alae peloderan. Guber- Chitwood, 1935). naculum small, trough-like. AP 9 : Length 2.6 Syn. Allantoneminae Pereira, 1931. mm. Swollen, worm-shaped; tail not coiled Allantonematidae (Pereira, 1931) when relaxed by heat, ventral side not turned Chitwood and Chitwood, 1937. outward. Tail with diagnostic spike; not dor- Contortylenchidae Ruhm, 1956. sally bent, papilliform. Stylet present. Uterus DIAGNOSIS (Emended): Sphaerulariidae. containing several eggs. Oviparous. FL 9 •. With or -without stylet. Without eggs HOST INSECT: in uterus. FL $ : With tylenchoid anterior end. Coleoptera; Cyllene. Stylet usually present, with or without knobs. TYPE SPECIES: Caudal alae usually peloderan when present, Aphelenchulus mollis Cobb, 1920. not voluminous. Spicules and gubernaculum This nematode needs to be recollected and tylenchoid, when present. AP 9 : Swollen, restuclied to obtain additional information on uterus not everted. its morphological relationships with various

Copyright © 2011, The Helminthological Society of Washington JANUARY, 1967] HELMINTHOLOGICAL SOCIETY 79 groups in the family. The shape of the AP ?, tail drawn in by expansion. Stylet absent. which aids in separating this genus from Con- Vulva with distinct lips; ovary and oviduct tortylenchiis, is shown in Fig. 1G. short; uterus occupies up to 85% of body. Ovoviviparous. Genus: Bovienema Nickle, 1963 HOST INSECTS: (Figs. IB, 3D, 4A) Coleoptera; Aphodius, Spondylis, Throscus. FL?: Small. Stylet well-developed. FLS: Hemiptera; Gerris, Velia, Nepa. Diptera; Stylet present. With peloderan caudal alae. Bibio. Gubernaculum present. AP2: Length 0.5- TYPE SPECIES: 0.75 mm. Body swollen, habit in the form Bradijnema rigidum (von Siebold, 1836) zur of a tight circle, with externally turned ventral Strassen, 1892. side, see Contortylenchiis. Tail tip character- Syn. Filaria rigida von Siebold, 1836. istically peg-like, directed dorsally. Stylet al- Allantonema rigida (von Siebold, ways present, well-developed, not retracted. 1836) Moniez, 1891. Intestine not differentiated. Oviparous. Gonad OTHER SPECIES: with 2 or 3 flexures, one anterior in neck B. bibionis Wachek, 1955. region, one posterior in vicinity of vulva; one B. gerridis Poisson, 1933. to three eggs in uterus at one time. B. nepal Poisson, 1933. HOST INSECT: B. strasseni Wiilker, 1923. Coleoptera; Ptiijogenes. B. trixagi Wachek, 1955. TYPE SPECIES: B. veliae Poisson, 1933. Bovienema tomici (Bovien, 1937) Nickle, 1963. Genus: Chondronema Christie and Syn. Aphelenchulus tomici Bovien, 1937. Chitwood, 1931 Contortylenchiis tomici (Bovien, (Figs. 1C, 4D) 1937) Ruhm, 1956. Head with 4 distinct papillae. Stylet small, OTHER SPECIES: tylenchoid. Dorsal esophageal gland present. Bovienema chalcographi (Fuchs, 1938) Amphidial openings lateral, slightly nearer n. comb. mouth than papillae; amphidial glands large. Syn. Parasitylenchus contortus chalco- Esophagus without bulb-like swelling. A pair graphi Fuchs, 1938. of large lateral pores on the tail. Male with Parasitylenchus contortus f. chalco- small caudal alae; without gubemaculum. graphi (Fuchs, 1938) W. Schnei- Testis reflexed. Body of female degenerat- der, 1939. ing into nearly structureless sac filled with Contortylenchiis chalcographi developing embryos. Body-cavity parasites (Fuchs, 1938) Riihm, 1956. throughout larval development, but free-living throughout adult stage. Genus: Bradijnema zur Strassen, 1892 HOST INSECT: (Figs. 1H, 3E, 4A) Coleoptera; Passalus. FL9 : With smooth cuticle. Lips faintly set TYPE SPECIES: off. Stylet absent. Stomal region faintly cutic- Chondronema passali (Leidy, 1852) Christie ularized. Esophageal glands well-developed, and Chitwood, 1931. extending almost to the genital area. Intestine Syn. Nematoideum passali Leidy, narrow-celled, used for storage. FLcJ: With 1852. smooth cuticle. Lips not distinctly set off. The life history of this nematode differs Stylet absent. Caudal alae absent or faintly from the rest of the genera in the the family developed in B. trixagi. Spicules tylenchoid. because only the larval stages are parasitic in Gubemaculum always present. AP $ : Cuticle the haemocoel of the insect. No AP 9 's could and hypodermis traversed by small narrow be found in the body cavity of the insect. canals. Metabolic products white. Head and Christie and Chitwood described the adult

Copyright © 2011, The Helminthological Society of Washington 80 PROCEEDINGS OF THE [VOL. 34, No. 1 forms as free-living, and found them in the Aphelenchulus contortus typographi frass of the beetles. Larval nematodes prob- (Fuchs, 1915) Micoletzky, 1925. ably leave the host during oviposition. There Anguillulina contortus typographi is no reference to the type of gonad in the (Fuchs, 1915) Baylis and Daub- female, but a small tylenchoid stylet is present. ney, 1926. The morphology and life history of the adult Aphelenchulus diplogaster (Fuchs, forms need more study. 1915) Filipjev, 1934. Tylenchus contortus cembrae Fuchs, Genus: Contortylenchusl(uhm, 1956 1915. (Figs. II, 3F, 4A) Parasitylenchus contortus cembrae FL 9 : Stylet usually well-developed, tip (Fuchs, 1915) Fuchs, 1929. symmetrical, lumen narrow, basal knobs vari- OTHER SPECIES: able, usually prominent. Small postuterine sac. Ovary with fewer cells than in neoparasity- C. acuminati Ruhm, 1956. lenchs; vulva posteriorly located, with small C. amitini Ruhm, 1956. opening, without projecting lips. FL $ : Stylet C. barbems (Massey, 1957) Ruhm, 1960. present, dimorphic. Testis with flexure. Tail Syn. Aphelenchuhis barbems Massey, bulkier, shorter than in neoparasitylenchs; 1957. rounded, always with peloderan caudal alae, C. brevicomi (Massey, 1957) Ruhm, 1960. usually narrow. Spicules small, tylenchoid. Syn. Aphelenchuhis brevicomi Massey, Gubernaculum small. AP 5 : Sulphur to brown- 1957. yellow. Always sausage-shaped or worm- C. cryphali Ruhm, 1956. shaped; often coiled when relaxed by heat, Syn. Contortylenchiis cryphali Ruhm, with ventral side always turned outward; the 1954. nomen nudum. "contortus" group of Fuchs. Body surface smooth, no body swellings; anterior end often C. cunicularii (Fuchs, 1929) Ruhm, 1956. wrinkled, narrower than rest of body in the Syn. Tylenchus contortus cunicularii form of a movable mouth cone with a stylet. Fuchs, 1929. Intestine retained, well defined from rest of Aphelenchulus cunicularii (Fuchs, tissue. Gonad may have 1 or 2 flexures; ovi- 1929) Filipjev, 1934. duct longer than in neoparasitylenchs; uterus Parasitylenchus contortus f. c-uni- narrow, not expanded to hypodermis; vulva cularii (Fuchs, 1929) W. Schnei- subterminal, deeply recessed; vagina heavily der, 1939. cuticularized. Anus functionless. Tail rounded, C. elongatus (Massey, 1960) Nickle, 1963. or tapering to nipple-like papilla. Oviparous. Syn. Aphelenchuhis elongatus Massey, HOST INSECTS: 1960. Coleoptera; Ips, Dendroctonus, Cryphalus, C. grandicolli (Massey, 1957) Ruhm, I960. Hylastes, Orthotomicus. Syn. Aphelenchulus grandicolli Massey, TYPE SPECIES: 1957. Contortylenchiis diplogaster (von Linstow, C. laricis (Fuchs, 1929) Rtihm, 1956. 1890) Ruhm, 1956. Syn. Tylenchus contortus laricis Fuchs, Syn. Allantonema diplogaster von Lin- 1929. stow, 1890. Aphelenchulus laricis (Fuchs, 1929) Tijlenchus diplogaster (von Linstow, Filipjev, 1934. 1890) Fuchs, 1915. Parasitylenchus contortus f. laricis Tylenchus contortus typography (Fuchs, 1929) W. Schneider, Fuchs, 1914. 1939. Parasitylenchus contortus typo- C. reversus (Thorne, 1935) Ruhm, 1956. graphi (Fuchs, 1915) Micoletzky, Syn. Aphelenchulus reversus Thorne, 1922. 1935.

C. spirits (Massey, 1957) Riihm, 1960. Genus: Howardula Cobb, 1921 Syn. Aphelenchulus spirus Massey, 1957. Syn. Tylenchinema T. 'Goodey, 1930. The contortylenchs have the type of ovary Prothallonema Chris- that is typical of the family and do not differ tie, 1938. markedly from the rest of the genera. There- Acarinocola Warren, fore, Contortylenchidae Riihm, 1956, is con- 1941. sidered a synonym of Allantonematinae. (Figs. 1L, 1M, 3J, 3K, 3L, 4A) Genus: Heterotylenchus Bovien, 1937 FL 9 : Stylet present. Lips faintly set off. (Figs. IE, 1J, 2N, 2O, 31, 5B) FL $: Stylet absent. Caudal alae narrow, peloderan. Gubernaculum present. AP 9 : Four distinct forms. With alternation of Length 1.2-7.0 mm. Stylet present. White, gamogenetic and parthenogenetic generations. even in old age. Head and tail retracted. FL 9 : Small, slender, about 0.5 mm long. Lips Oviparous or ovoviviparous. Ovary and ovi- faintly set off. Stylet present, with knobs. duct in oviparous species coiled within body; Esophagus with overlapping glands and indis- in ovoviviparous species, crowded together in tinct orifices. Anus vestigial. Vulva posterior, anterior section of body. Vulva posterior, al- close to anus. FL<5: Slender, about 0.5 mm most terminal, lips protrude slightly. long. Stylet present, small, knobbed. Tail HOST INSECTS AND MITES: conical. Caudal alae absent. Spicules usually Coleoptera; Diabrotica, Phyllotreta. Dip- small, tylenchoicl. Gubernaculum lacking or tera; Oscinella. Thysanoptera; Aptinothrips, very small. Gamogenetic female (AP9): Taeniothrips, Frankliniella. Acarina; Para- Yellow-brown. Body swollen, curved, inert, situs, Poecilochirus, Haemogamasiis, Eury- much larger than the free-living stage (up to parasitus, Cosmolaelaps, Acarinocola. 3.00 mm); both ends rounded; or, tail with TYPE SPECIES: spike-like papilla. Stylet present, may be Howardula benigna Cobb, 1921. retracted into body. Intestine a syncytium. OTHER SPECIES: Anus vestigial. Ovary, oviduct and uterus of H. acarinorum Wachek, 1955. only moderate dimensions, outstretched; or, H. aoronymphium Welch, 1959. ovary and oviduct may lie in many convolu- H. aptini. (Sharga, 1932) Wachek, 1955. tions. Oviparous. Eggs larger than those of Syn. Tylenchus aptini Sharga, 1932. parthenogenetic female. Parthenogenetic fe- AngiiilluUna aptini (Sharga, 1932) male: More slender (up to 1.00 mm long). Lysaght, 1936. Anterior end drawn in by expansion. Tail H. clamger (Warren, 1941) Wachek, 1955. conical or dome-shaped. Stylet weakly de- Syn. Acarinocola claviger Warren, 1941. veloped. Esophagus degenerate. Ovary greatly H. cuneifer (Warren, 1941) Wachek, 1955. developed, reaching into head region. Ovip- Syn. Acarinocola cuneifer Warren, 1941. arous or ovoviviparous; when ovoviviparous, H. dubhim (Christie, 1938) Nickle, 1965. ovary and oviduct displaced anteriorly by Syn. Prothallonema dubhim Christie, uterus. 1938. HOST INSECTS: H. hirsuta (Warren, 1941) Wachek, 1955. Diptera; Hylemya, Musca. Coleoptera; Bem- Syn. Acarinocola hirsutus Warren, 1941. bidion, Clivina. Siphonaptera; Coptopsylla, H. oscinellae (Goodey, 1930) Wachek, Ceratophyllus. 1955. TYPE SPECIES: Syn. Tylenchinema oscinellae Goodey, Heterotylenchiis aberrans Bovien, 1937. 1930. OTHER SPECIES: H. phyllotretae Oldham, 1933. H. bovieni Wachek, 1955. H. terribilis (Warren, 1941) Wachek, 1955. H. stammeri Wachek, 1955. Syn. Acarinocola terribilis Warren, 1941. H. ivillkeri Wachek, 1955. This genus contains several diverse forms, PI. pawloivskyiKumchkin, 1960. and after more study it may be separated into

Copyright © 2011, The Helminthological Society of Washington 84 PROCEEDINGS OF THE [VOL. 34, No. 1 valid genera or subgenera. The distinguishing well-developed, with basal swellings. Ovary feature is the lack of stylet in the male. with more cells than Contortylenchus and Allantonema spp. FL 6 : Stylet present, not Genus: Metaparasitylenchus prominent. Caudal alae peloderan or lepto- (Wachek, 1955) n. grad. deran. Spicules and gubernaculum small, (Figs. 2S, 2T, 3N, 4A) tylenchoicl. AP 5 : Swollen, slightly mobile, Larvae leave host after second moult, third usually sausage- or worm-shaped. Stylet not moult occurs in free-life. After copulation, im- retracted into inner body. Tail terminal with pregnated young females infect host larva or or without papilla. Oviparous or ovoviviparous. pupa, moult once in new host for fourth and Ovary and oviduct intertwined irregularly, last time (after Wachek, 1955). FL?: Stylet crowded together anteriorly; uterus long, wide, strongly knobbed. FL $ : Stylet present. Cau- filled with eggs or larvae. dal alae pelocleran, frequently broad. Guber- HOST INSECTS: naculum present. AP 9 : White, seldom with Coleoptera (bark beetles); Hylurgus, Hy- yellow-brown deposits. Oviparous or ovovivip- lastes, Dendroctonus, Scolytus, Cryphalus, arous. Crypturgus, Pityophthorus, Pityogenes, Ips. HOST INSECTS: TYPE SPECIES: Coleoptera (not bark beetles); Cryptophagus, Neoparasitylenchus cnjphali (Fuchs, 1914) Mijcetophagus, Telmatophilus, Cossonus, Te- 11. comb. tropium, Strangalia, Rhizophagus, Plelmis, Syn. Tylenchus dispar cnjphali Fuchs, Latelmis, Riohis. 1914. TYPE SPECIES: Parasitylenchus dispar var. cnjphali Metaparasitylenchus telmatophili (Wachek, (Fuchs, 1914) Micoletzky, 1922. 1955) n. grad. Aphelenchuhis cnjphali (Fuchs, Syn. Parasitylenchus (Metaparasitylen- 1914) Filipjev, 1934. chus} telmatophili Wachek, Parasitylenchus dispar f. cnjphali 1955. (Fuchs, 1914) Schneider, 1939. OTHER SPECIES: Parasitylenchus (Parasitylenchus) M. cossoni (Wiilker, 1929) n. comb. cnjphali (Fuchs, 1914) Ruhm, Syn. P. cossoni Walker, 1929. 1956. P. (M.) cossoni (Wiilker, 1929) OTHER SPECIES: Wachek, 1955. N. avulsi (Massey, 1958) n. comb. M. cryptophagi (Wachek, 1955) n. comb. Syn. P. (M.) cryptophagi Wachek, 1955. Syn. Parasitylenchus avulsi Massey, 1958. M. helmidi-s (Wachek, 1955) n. comb. P. (P.) avulsi (Massey, 1958) Syn. P. (M.) helmidis Wachek, 1955. Ruhm, 1960. M. mycetophagi (Wachek, 1955) n. comb. N. betulae (Rtihm, 1956) n. comb. Syn. P. (M.) mycetophagi Wachek, 1955. Syn. P. (P.) betulae EiT^m, 1956. N. chalcographi (Fuchs, 1938) n. comb. M. oschei (Rtihm, 1956) n. comb. Syn. P. dispar chalcographi Fuchs, 1938. Syn. P. (M.) oschei Rtihm, 1956. P. dispar f. chalcographi (Fuchs, M. rhizophagi (Wachek, 1955) n. comb. 1938) Schneider, 1939. Syn. P. (M.) rhizophagi Wachek, 1955. P. chalcographi (Fuchs, 1938) M. strangalioe (Wachek, 1955) n. comb. Filipjev & Sch. Stek., 1941. Syn. P. (M.) strangaliae Wachek, 1955. P. (P.) chalcographi (Fuchs, 1938) M. tetropii (Wachek, 1955) n. comb. Ruhm, 1956. Syn. P. (M.) tetropii Wachek, 1955. N. cinerei (Fuchs, 1929) n. comb. Syn. Tylenchus dispar cinerei Fuchs, Genus: NeoparosUylenchusn. gen. 1929. (Figs. 2Q, 3P, 4A) Aphelenchuhis cinerei (Fuchs, 1929) With 3 distinct forms; one free-living sexual Filipjev, 1934. generation, one swollen female developing from P. dispar f. cinerei (Fuchs, 1929) the mated free-living female. FL9: Stvlet Schneider, 1939.

P. (P.) cinerei (Fuchs, 1929) N. louelkeri (Ruhm, 1956) n. comb. Ruhm, 1956. Syn. P. (P.) wuelkeri'Ruhm, 1956. P. dispar pusilli Fuchs, 1938. N. xylebori (Schvester, 1950) n. comb. P. dispart, pusilli (Fuchs, 1938) Syn. P. dispar xylebori Schvester, 1950. Schneider, 1939. P. (P.) xylebori (Schvester, 1950) P. pusilli (Fuchs, 1938) Filipjev Ruhm, 1956. and Stek., 1941. N. hi/last is (Wiilker, 1923) n. comb. Genus: ParasitylenchoidesWachek, 1955 Syn. Tylenchus hylastis Wiilker, 1923. (Figs. 2U, 3S, 4A) Tylenchus dispar ateri Fuchs, 1929. FL5 : Excretory pore 0.105-0.125 mm from Tylenchus dispar cunicularii Fuchs, anterior end, against 0.038-0.072 mm in Neo- 1929. para.sitylenchus (Diagnostic). Stylet present, P. hylurgi Filipjev, 1934. with knobs. FL $ : Stylet present. Caudal P. dispar f. cunicularii (Fuchs, 1929) alae peloderan or absent. Spicules tylenchoid. Schneider, 1939. Gubernaculum small. APS: Swollen, worm- P. dispar L ateri (Fuchs, 1929) shaped; head and tail not drawn in by expan- Schneider, 1939. sion; with mouth-cone and tail papilla. Stylet P. hylastis (Wiilker, 1923) Filipjev, in normal position or retracted. Oviparous or 1934. ovoviviparous. With oviparity, ovary and ovi- P. (P.) hylastis (Wiilker, 1923) duct show few windings. With ovoviviparity, Ruhm, 1956. ovary displaced anteriorly or to the side as N. ligniperdae (Fuchs, 1929) n. comb. uterus fills 80% of body. Metabolic products Syn. Tylenchus ligniperdae Fuchs, 1929. yellow-brown. P. ligniperdae (Fuchs, 1929) HOST INSECTS: Filipjev, 1934. Coleoptera; Anobium, Plegaderus, Micro- P. (P.) ligniperdae (Fuchs, 1929) malthus, Ditoma, Paederus, Sciodrepa, Aleo- Ruhm, 1956. chara, Oxytelus, Stenus, Paromalus. N. orthotomici (Ruhm, 1960) n. comb. TYPE SPECIES: Syn. P. (P.) orthotomici Ruhm, 1960. P. steni Wachek, 1955. N. ovaritis (Massey, 1958) n. comb. OTHER SPECIES : Syn. P. ovarius Massey, 1958. P. anobii, Wachek, 1955. N. pessoni (Ruhm, 1957) n. comb. P. diatomae Wachek, 1955. Syn. P. (P.) pessoni Ruhm, 1957. P. korneri Wachek, 1955. N. pityophthori (Ruhm, 1956) n. comb. P. paederi Wachek, 1955. Syn. P. (P.) pttyophthoriJtijhm, 1956. P. paromali Wachek, 1955. N. poligraphi (Fuchs, 1938) n. comb. P. rheocharae Wachek, 1955. Syn. P. dispar poligraphi Fuchs, 1938. P. sciodrepae Wachek, 1955. P. dispar f. poligraphi (Fuchs, 1938) P. wichmanni Wachek, 1955. Schneider, 1939. P. poligraphi (Fuchs, 1938) Filipjev Genus: Parasitylenchus Micoletzky, 1922 andSch. Stek., 1941. Syn. Polymorphotylenchus P. (P.) poligraphi (Fuchs, 1938) Ruhm, 1956. Ruhm, 1956. (Figs. 2V, 2X, 2Y, 30, 4C) N. rugulosi (Schvester, 1957) 11. comb. With 5 distinct forms, 2 sexual generations, Syn. P. dispar rugulosi Schvester, 1957. one swollen female developing from mated P. (P.) rugulosi (Schvester, 1957) free-living female. Small parasitic female: Ac- Ruhm, 1960. tive, narrow, sausage-shaped, present in large N.scolyti (Oldham, 1930) n. comb. numbers; or motionless, yellow-brown, raisin- Syn. P. scolyti Oldham, 1930. shaped. Stylet present, may be retracted, P. secundus Fuchs, 1933. smaller than in large female; basal knobs P. (P.) scolyti (Oldham, 1930) small, flat. Gonad with a short flexure, uterus Ruhm, 1956. with few eggs; or greatly developed with nu-

Copyright © 2011, The Helminthological Society of Washington 86 PROCEEDINGS OF THE [VOL. 34, No. 1 merous flexures of ovary. Small parasitic male: tis (Fuchs, 1914) Micoletzky, Active, narrow. Lips not set off. Excretory 1925. pore opens near stylet. Stylet present, weakly Anguillulina dispar var. curvidentis developed. Esophagus and intestine weak. (Fuchs, 1914) Baylis and Daub- Gonad outstretched, extends almost to stylet. ney, 1926. Tail plump, rounded. Caudal alae present or Aphelenchulus curvidentis (Fuchs, absent. Spicules and gubernaculum small. Free- 1914) Filipjev, 1934. living stages not described. AP 9 : Swollen, Parasitylenchus dispar f. curvidentis sausage-shaped, inwardly curved, milky white (Fuchs, 1914) Schneider, 1939. with red-brown spots; may be transparent or Polyrnorphotylenchus (Thylakolen- partly dark brown. Stylet present, knobs very chus) curvidentis (Fuchs, 1915) small, flat. Gonad well-developed; ovary and Riihm, 1956. oviduct with numerous crowded loops in an- Parasitylenchus diplogenus Welch, 1959. terior end; uterus large; or gonad weakly de- Syn. Polyrnorphotylenchus diplogenus veloped with more or less long flexure, not (Welch, 1959) Baker, 1962. pushed anteriorly, uterus narrow with few Welch (1959) pointed out that Riihm im- eggs. properly used the type species of the genus HOST INSECTS: Parasitylenchus Micoletzky, 1922, as the type Coleoptera; Ips, Pityokteines. Diptera; Dro- species of his new genus Polyrnorphotylenchus. sophila. Filipjev's 1934 designation of Tylenchus dispar TYPE SPECIES: typographi Fuchs, 1915 as the only synonym Parasitylenchus dispar (Fuchs, 1915) Filip- of his type species, Parasitylenchus dispar jev, 1934. (Fuchs, 1915) may not have been the best Syn. Tylenchus dispar typographi Fuchs, choice; however, it is valid according to the 1914. nomen nudum. International Rules of Zoological Nomencla- Tylenchus dispar typographi Fuchs, ture. Therefore, most of the other species have 1915. been removed from Parasitylenchus and placed Parasitylenchus dispar var. typo- in a new genus Neoparasitylenchus. The graphi (Fuchs, 1915) Micoletzky, subgenera Sulphuretylenchus, Metaparasitylen- 1922. chus, and Proparasitylenchus have been raised Aphelenchulus dispar var. tijpo- to generic level. Polymorphotylenchus has graphi (Fuchs, 1915) Micoletzky, been synonymized with Parasitylenchus. 1925. Genus: Proparasitylenchus Wachek, Anguillulina dispar var. tijpographi 1955n. grad. (Fuchs, 1915) Baylis and Daub- ney, 1926. (Figs. 2P, 3R, 4A) Parasitylenchus dispar f. typographi FL 9 : Copulated females penetrate into (Fuchs, 1915) Schneider, 1939. host pupa. Stylet present, usually with strong Polymorphotylenchus (Polymorpho- knobs, never bare or cleft. FL $ : Stylet pres- tylenchus) typographi (Fuchs, ent. Caudal alae peloderan or leptoderan, 1915) Riihm, 1956. always present. Gubernaculum present. AP 9 : OTHER SPECIES: White, or faintly citron-yellow, never brown. Parasitylenchus curvidentis (Fuchs, 1914) Ovoviviparous. Larvae leave host after second Micoletzky, 1922. moult, two moults in free-life. Syn. Tylenchus dispar curvidentis Fuchs, HOST INSECT: 1914. Coleoptera (staphylinids); Platystethus, Me- Parasitylenchus dispar var. curvi- don, Trogophloeus, Zyras, Atheta, Oxytelus. dentis (Fuchs, 1914) Micoletzky, TYPE SPECIES: 1922. Proparasitylenchus platystethi (Wachek, Aphelenchulus dispar var. curviden- 1955) n. grad. '

Syn. Parasitylenchus (Proparasitylenchus) basal swellings flat, not knobbed. Esophagus platystethi Wachek, 1955. and intestine degenerate. Ovoviviparous. Vulva OTHER SPECIES: and anus difficult to see in mature parasites. P. athetae (Wachek, 1955) n. grad. Gonad almost fills body, flexed about 3 times. Syn. P. (P.) athetae Wachek, 1955. Papillate tail tip absent. P. boopini (Wachek, 1955) n. grad. HOST INSECTS: Syn. P. (P.) boopini Wachek, 1955. Coleoptera (bark beetles); Pityogenes, Ips, P. medonis (Wachek, 1955) n. grad. Polygraphus, Dryocoetes, Hylastes, Scolytus. Syn. P. (P.) medonis Wachek, 1955. TYPE SPECIES: P. myrmedoniae (Wachek, 1955) n. grad. Sulphuretylenchus sulphureus (Fuchs, 1938) Syn. P. (P.) myrmedoniae Wachek, n. grad. 1955. Syn. Parasitylenchus sulphureus chalco- P. oxyteli (Wachek, 1955) n. grad. graphi Fuchs, 1938. Parasitylenchus (Sulphuretylenchus) Syn. P. (P.) oxyteli Wachek, 1955. sulphureus (Fuchs, 1938) Riihm, P. trogophloei (Wachek, 1955) n. grad. 1956. Syn. P. (P.) trogophloei Wachek, 1955. OTHER SPECIES: Genus: Protylenchus Wachek, 1955 S. elongatns (Massey, 1958) n. comb. (Figs. IK, 3T, 4A) Syn. P. elongatus Massey, 1958. S. escherichi (Riihm, 1956) n. grad. FL 9 : Lips not set off. Stylet present. Syn. P. (S.) escherichi Riihm, 1956. FL $ : Lips not set off. Stylet well-developed S. fuchsi (Fuchs, 1938) n. grad. with three cleft knobs. Caudal alae absent. Syn. P. sulphureus poligraphi Fuchs, Spicules tylenchoid. Gubernaculum proximally 1938. thickened. AP 9 : Cuticle bare, hypodermis P. (S.) fuchsi (Fuchs, 1938) Riihm, thick. Stylet present, retracted within body. 1956. Tail papilla present. Ovary and oviduct long, S. grossmannae (Riihm, 1954) n. grad. coiled profusely throughout body; vulval lips Syn. Paratylenchus (Sulphuretylenchus) slightly arched forward; uterus short. Ovip- grossmannae Riihm, 1954. Lapsus arous. Metabolic products white. No alterna- calami. tion of generations. S. kleinei (Riihm, 1956) n. grad. HOST INSECT: Syn. Parasitylenchus (Sulphuretylenchus) Coleoptera; Heterocerus. TYPE SPECIES: kleinei Riihm, 1956. P. heteroceri Wachek, 1955. S. pilifronis (Massey, 1958) n. comb. Syn. P. pilifronis Massey, 1958. Genus: Sulphuretylenchus Riihm, The significance of a group of genera with 1956 n. grad. a long stylet, without knobs, and with a dorsal (Figs. 2R, 3X, 4A) gland orifice which opens over a stylet length FL9: Longer than most sphaerulariids. behind the stylet base, is not known to me at V—A distance longer than neoparasitylenchs. this time. It includes some species of Sul- Excretory pore opens below nerve ring. Stylet phuretylenchus, Howardula, Heterotylenchiis large; lumen wide. FL $ : Stylet present. Tail and others. Future taxonomic work \vill un- longer than typical neoparasitylench. Caudal doubtedly clarify this relationship. alae peloderan. Spicules and gubemaculum small. AP9: Shape variable, usually long, Genus: Scatonema Bovien, 1932 with swollen spot on tube-shaped body, swell- (Figs. 2AA, 3U, 4A) ing caused by congestion of larvae and eggs FL9 : Small (0.33 mm long), slender. Sty- (Endotokie matricide). Body surface char- let well-developed, with basal knobs; tip with acteristically wavy, with constrictions. Sulphur oblique cut. First part of esophagus with dis- to yellow-brown or dark brown. Stylet re- tinct cuticular lining; two large ventral esoph- tracted into inner body, spear tip asymmetrical, ageal glands open separately into esophagus in

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Copyright © 2011, The Helminthological Society of Washington 90 PROCEEDINGS OF THE [VOL. 34, No. 1 front of nerve ring. No anal opening found. DIAGNOSIS (Emended): Sphaerulariidae. Oocytes few. Vagina surrounded by circle of FL9: Found in Eucalyptus gall. Eggs in large perivaginal cells. Tail tapering poste- uterus. Stylet well-developed. FL $ : With ty- riorly. FL6: Small (0.33 mm long), slender. lenchoid anterior end. Stylet present, knobbed. Stylet absent. Esophageal glands not seen. Caudal alae peloderan, not voluminous. Spic- Tail tapering behind anus. Spicules and guber- ules tylenchoid. Gubernaculum absent. AP 9 : naculum tylenchoid. Caudal alae present, nar- Swollen, uterus not everted. row. AP9 : Cigar- or sausage-shaped. Often coiled. Ovary greatly developed; swelling at Genus: Fergitsobia Currie, 1937 end of oviduct serves as receptaculum seminis; Syn. Angutttulma (Fergits- uterus occupies bulk of body; perivaginal cells obia) Currie, 1937. strikingly developed. Ovoviviparous. (Figs. ID, 3H, 4B) HOST INSECT: Three stages found: FL9: In gall. Head Diptera; Scatopse. offset, narrower than body, which is quite TYPE SPECIES: plump and faintly annulated. Spear tylenchoid Scatonema iviilkeri Bovien, 1932. with well-developed basal knobs. Anterior part of esophagus distended by ampullae filled with Genus: Dotylaphus Andrassy, 1958 esophageal secretions and appearing valve-like (Fig.3G) at times; esophagus narrows considerably at Based on one specimen. FL9 : Cuticle an- isthmus surrounded by nerve ring; glands nulated. Head not set-off, with small lips and overlap the intestine; dorsal gland orifice papillae. Stylet short, well-developed, similar typically tylenchoid, just behind spear base; to species of Dorylaimus, however, tip slanting subventral esophageal glands empty into swol- ventrally; guiding ring asymmetrical; stylet len median bulb-like area. Excretory pore pos- knobs are lacking; stylet continuation sclero- terior to nerve ring. Female gonad prodelphic; tized, relatively long; lumen wide (similar to no postuterine sac; vulva with distinct lips. Dorylaimus). Esophagus without muscles, Eggs present. Rectum and anus visible. FL 6 : without bulb, opening gradually into intes- Spicules paired, robust, knobbed, arcuate. tine; esophageal glands well-developed, long, Gubernaculum absent. Goodey illustrates a lie free in body cavity, outstretched, with en- narrow, peloderan caudal alae. Stylet present. larged ampulla-like openings; subventral gland Gonad single, outstretched. Tails of both sexes orifices in the middle of esophagus; dorsal short, conoid, rounded. AP 9 : Similar in shape gland orifice between anterior end and middle to Heterotijlenchus from carabid beetles. Ovip- of esophagus, far behind stylet. Lateral organs arous. Ovary winds several times within body. not seen. Lateral field well-developed, with Stylet retracted. Numerous lateral somatic several lines. Gonad single, prodelphic; vulva setae present. is posteriorly located. Tail short, rounded. HOST INSECT: FL5: Unknown. AP9: Unknown. Diptera; Fergusonina. HOST INSECT: TYPE SPECIES: Unknown. Fergusobia tumifacien.s (Currie, 1937) TYPE SPECIES: Chitwood and Chitwood, 1950. Dotylaphus riihmi Andrassy, 1958. Syn. Anguillulina (Fergusobia) tumi- This specimen belongs in this family. Un- faciens Currie, 1937. fortunately, it can fit into a few existing genera Anguillulina (Fergusobia) curriei and without further information it is difficult (Currie, 1937) Johnston, 1938. to place at this time. Therefore, these taxa I have studied males and females from the should be treated as genus and species in- gall and also the adult parasitic females from quirendae. the body cavity of the insect. Though the sub- SUBFAMILY: ventral gland orifices are swollen in the middle Fergusobiinae J. B. Goodey, 1963. of the esophagus, no criconematid valve could Syn. Fergusobiidae Siddiqi and J. B. be found. This fact dismisses any possible Goodey, 1963. association with the Criconematidae and dis-

Copyright © 2011, The Helminthological Society of Washington JANUARY, 1967] HELMINTHOLOGICAL SOCIETY 91 allows the family Fergusobiidae Siddiqi and OTHER SPECIES: J. B. Goodey, 1963. Fisher (1965) has re- I. fungorum (Biitschli, 1873) Fil. and Stek., cently found that the female in the gall pro- 1941. duces eggs. This separates this genus from all Syn. Tylenchus fungo-rum Biitschli, 1873. other members of this family. Hexatylus fungomm (Biitschli, SUBFAMILY: 1873) T. Goodey, 1932. lotonchiinae T. Goodey, 1953. Tylenchus (Anguillulina) fungorum Syn. lotonchiidae (T. Goodey, 1953) (Biitschli, 1873) Filipjev, 1934. Skarbilovich, 1959. Neotylenchus fungorum (Biitschli, DIAGNOSIS (Emended): Sphaerulariidae. 1873) Filipjev, 1936. FL 9 : With post-vulval and mid-ventral sup- Anguillulina fungorum (Biitschli, plement. Without eggs. Stylet well-developed. 1873) W. Schneider, 1939. FL $ : Head bilaterally symmetrical, tri-lobed, I. bifurcatum T. Goodey, 1953. flattened dorso-ventrally. Stylet not well- /. cephalostrictum Meyl, 1954. developed. Spicules large, L-shaped, elaborate. I. macrospiculatum (Meyl, 1954) J. B. Caudal alae voluminous, envelop tail tip. Post Goodey, 1956. anal papillae sometimes present. AP 5 : Un- Syn. Hexatylus macrospiculatus Meyl, known. 1954. I. mycophilum Meyl, 1954. Genus: lotonchium Cobb, 1920 T. Goodey (1953) and J. B. Goodey (1956, (Fig.3M) 1963) showed the correct sphaerulariicl status Esophagus ill-defined; dorsal gland orifice of lotonchium. I also agree with the synonymy joining esophageal lumen 4 head widths from of lotonchiidae (T. Goodey, 1953) Skarbilo- anterior end of body; subventral esophageal vich, 1959, with lotonchiinae T. Goodey, 1953. glands opening about 4—5 head-widths further In so doing we may discount Biitschli's female back; anterior part of esophagus cylindrical, of Tylenchus imperfectus (His Fig. 7d) which nerve ring crossing slightly constricted isthmus; showed an egg in the female, and accept the posterior part not distinguishable from intes- sphaerulariid ovary as described by T. Goodey. tine. Lateral fields, deirids and hemizonid However, the female with the egg may be present in both sexes. Female: Spear well- valid and could represent a situation similar to developed, probably changing shape at final that of Fergusobia. The life cycles of these moult. Male: Head offset by expansion, stylet nematodes need more study, especially the not well-developed. Spicules large, markedly identity of the host insects, if any, and the angular and L-shaped, sometimes with pos- descriptions of AP 2 's. terior extensions protruding through cloaca. Gubernaculum absent. Pre-adult male larva SUMMARY precocious; having fully developed spicules A taxonomic revision of the insect parasitic and gonad but no caudal alae. Head of larval nematode family Sphaerulariidae Lubbock, male radially symmetrical, changing to adult 1861 is presented and includes 4 subfamilies, shape at final moult. AP9: Unknown. 21 genera, and 116 species. The family group HOST INSECT: diagnoses are emended, and generic diagnoses Unknown. Found in basidiomycetous fungi are given along with a listing of species and Entoloma, Pleurotiis, Hygrophorus, and Tri- synonymies. The dorsal esophageal gland ori- choloma. fice of Sphaerularia bombi was found for the TYPE SPECIES: first time and was located in the normal lotonchium imperfect-urn (Biitschli, 1876) tylenchoid position. This discovery required Cobb, 1920. the shifting of the Sphaerulariidae from the Syn. Tylenchus imperfectus Btitschli, Aphelenchoidea to the Tylenchoidea, and be- 1876. cause the Sphaerulariidae is an older family Anguillulina (lotonchium) imper- group name than the Allantonematidae the fecta (Btitschli, 1876) W. Schnei- latter drops to subfamily status. Polymorpho- der, 1939. tylenchus Riihm, 1956 is synonymized with

Parasitylenchus Micoletzkv, 1922 and P. dispar , AND M. B. CHITWOOD. 1937. An Intro- (Fuchs, 1915) Filipjev, 1934 is considered as duction to Nematology. Baltimore, Maryland. the type species of Parasitylenchus. The re- CHRISTIE, J. R. 1938. Two nematodes associated with decaying citrus fruit. Proc. Helminth. maining species of the old genus Parasitylen- Soc. of Wash. 5(1): 29-33. chus are placed in a new genus Neoparasity- , AND B. G. CHITWOOD. 1931. Chondro- lenchns, requiring 16 new combinations. The nema passali (Leidy, 1852) n. g. (Nematoda) families Contortylenchidae and Allantonemati- with notes on its life history. Jour. Wash. dae are considered at this time to be synonyms Acad. Sci. 21: 356-364. of the Allantonematinae. Fergusobiidae is COBB, N. A. 1920. One hundred new nemas. synonymizecl with Fergusobiinae which is Contrib. Sci. Nematol. 9: 217-343. placed as a subfamily of the Sphaerulariidae. . 1921. Howardula benigna, a nemic parasite of the cucumber beetle (Diabrotica). The writer agrees with Goodey's synonymy of Contrib. Sci. Nematology 10: 345-352. lotonchiidae with lotonchiinae and its place- CURRIE, G. A. 1937. Galls on Eucalyptus trees. ment with these insect parasites. Dotylaphus A new type of association between flies and ruhmi Andrassy, 1958 is treated as genus and nematodes. Proc. Linn. Soc. N.S. Wales 62 species inquirendae. The subgenera Sulphur- (3/4): 147-174. etylenchus Riihm, 1956, Metaparasitylenchus DUFOUR, L. 1837. Recherches sur quelques en- Wachek, 1955, and Proparasitylenchus Wachek, tozoaires et larves parasites des insectes or- 1955 are raised to generic rank. Host insects thopteres et hymenopteres. Ann. Sci. Natur. are given whenever possible because they are Zool. 7: 5-20. FILIPJEV, I. N. 1934. Harmful and useful nema- useful in identification. Typical life cycles are todes in rural economy. 440 pp. Leningrad, illustrated along with plates of representative Moskva. adult parasitic females, and free-living male , AND J. H. SCHUURMANS-STEKHOVEN, JR. and female stages. 1941. A manual of agricultural helminthology. E. J. Brill, Leiden, 878 pp. LITERATURE CITED FISHER, J. M. 1965. (Personal Communication). FUCHS, G. 1914. Tijlenchus dispar cnrvidentis ANDRASSY, I. 1958. Erd-und Siisswassernemato- m. und Tijlenchus dispar cryphali m. Zool. den aus Bulgarian. Acta Zoologica, Acad. Sci. Anz. 45(5): 195-207. Hung. IV (1/2): 1-88. . 1915. Die Naturgeschichte der Nema- BAKER, A. D. 1962. Check lists of the nematode toden und einiger anderer Parasiten des Zp.s superfamilies Dorylaimoidea, Rhabditoidea, typographies L. und Hylobius abietif, L. Zool. Tylenchoidea, and Aphelenchoidea. E. J. Brill, Jahrb. (Syst.) 38(3/4)': 109-222. Leiden, 261 pp. . 1929. Die Parasiten einiger Riissel- und BOVIEN, P. 1932. On a new nematode, Sca- Borkenkjifer. Ztschr. Parasitenk. 2(2): 248- tonema ivi'dkeri gen. et spec. n. parasitic in the 285. body-cavity of Scatopse fuscipes Meig. (Dip- . 1933. Einige Nematoden bei Scolytus tera Nematocera). Vid. Meddel. fra Dansk scoltjtus. F. Capita Zoologica 4(1): 1—45. naturh. Forening. K0benhavn. 94: 13-32. 1938. Neue Parasiten und Halbpara- •. 1937. Some types of association be- siten bei Borkenkafern und einige andere tween nematodes and insects. Vid. Meddel. Nematoden. II, III, IV. Zool. Jahrb. (Syst.) fra Dansk naturh. Forening. K0benhavn 101: 71(1/2): 123-190. 1-114. GOODEY, J. B. 1956. Observations on species of 1944. Proatractonema sciarae n. g., n. the genus, lotonchium Cobb, 1920. Nema- sp., a parasitic nematode from the body cavity tologica 1(3): 239-248. . 1960. The classification of the Aphelen- of a dipterous larva. Vid. Meddel. fra Dansk choidea Fuchs, 1937. Nematologica 5: 111- naturh. Forening. K0benhavn. 108: 1-14. 126. BUTSCHLI, O. 1873. Beitrage zur Kenntnis der 1963. Soil and freshwater nematodes. freilebenden Nematoden. Nova Acta (Acad. (Revision of T. Goodey, 1951) Methuen, Nat. Curios) 36: 144 pp. London, 544 pp. . 1876. Untersuchungen iiber freilebende GOODEY, T. 1930. On a remarkable new nema- Nematoden und die Gattung Chaetonotus. tode, Tylenchinema oscinellae gen. et sp. n., Ztschr. Wissensch. Zool. 26: 363-413. parasitic in the frit fly, Oscinella frit L., at- CHITWOOD, B. C. 1935. Nomenclatorial notes, 1. tacking oats. Philos. Tr. Roy. Soc. London Proc. Helminthol. Soc. of Wash. 2: 51-54. 218: 315-343.

. 1953. On certain eelworms, including . 1963b. Bovienema (Nematoda: Allan- Biitschli's Tylenchus fungorum, obtained from tonematidae), a new genus parasitizing bark toadstools. Jour. Helminth. XXVII (1/2): 81- beetles of the genus Pityogenes Bidel, with 94. notes on other endoparasitic nematod.es of KHAN, M. A. 1957a. Sphaerularia bombi Duf. scolytids. Proc. Helminthol. Soc. Wash. 30 {Nematoda: Allantonematiclae) infesting bum- (2): 256-262. blebees and Sphaerularia hastata sp. nov. in- -, AND G. W. WOOD. 1964. Howardula festing bark beetles in Canada. Canad. Jour. aptini (Sharga, 1932) parasitic in blueberry Zool. 35(4): 519-523. thrips in New Brunswick. Can. Jour. Zool. . 1957b. Sphaerularia ungulacauda sp. 42(5): 843-846. nov. (Nematoda: Allantonematidae) from the PEREIRA, C. 1931. Myenchus botelhoi n. sp., Douglas fir beetle, Dendroctonus pseudotsugae curiosa nematoide parasite de Limnobdella Hopk., with key to Sphaerularia species brasiliensis Pinto (Hirudinea). Thesis, Fac. (Emended). Canad. Jour. Zool. 35(6): 635- Med., Sao Paulo, 30 pp. 639. POINAR, G. O. 1965. The bionomics and para- 1960. Stictylus hastatus (Khan, 1957) sitic development of Tripins sciarae (Bovien) n. comb., and Stictylus ungidacaudus (Khan, (Sphaerulariidae: Aphelenchoidea), a nema- 1957) n. comb. (Nematoda: Neotylenchidae). tode parasite of sciarid flies (Sciaridae: Dip- Canad. Jour. Zool. 38(1): 225-226. tera). Parasitology 55: 559-569. KUROCHKIN, Y. V. 1960. The nematode Hetero- POISSON, R. 1933. Trois nouvelles especes de tylenchus pawlowskyi n. sp. castrating flea nematodes cle la cavite generale d'hemipteres carriers of plague. Dokladi Academii Nauk aquatiques. Ann. Parasit. Humaine et Com- U.S.S.R. 135(5): 1281-1284. pare. 11(6): 463-466. LEUCKART, R. 1884. Ueber einen neuen hetero- ROY, D. N., AND P. K. MUKHERJEE. 1937. Al- genen Nematoden. Tagebl. 57. Vers. Deut- lantonema stricklandi n. sp., a parasitic nema- scher Naturf. u. Aerzte. p. 320. tode of houseflies, Mnsca vicina. Ann. Trop. LINSTOW, O. VON. 1890. Uber Allantonema und Med. and Parasit. 31(4): 453-456. Diplogaster. Zentralbl. Bakt. und Paraskde RiiHM, W. 1956. Die Nematoden der Ipiden. 8(16): 489-493. Parasitologische Schriftenreihe 6: 1-437. VEB LUBBOCK, J. 1861. On Sphaerularia bombi. Nat. Fischer Verlag, Jena. Hist. Rev. 1(1): 44-57. . 1960. Ein Beitrag zur Nomenklatur und MASSEY, C. L. 1956. Nematode parasites and Systematik einiger mit Scolytiden vergesell- associates of the Engelmann spruce beetle schafteter Nematodenarten. Zool. Anz. 164 (Dendroctonus engelmanni). Proc. Helmin- (5/6): 201-213. thol. Soc. Wash. 23(1): 14-24. . 1957. Four new species of Aphelen- SCHNEIDER, W. 1939. Wurmer oder Vermes. II. chulus (Nematoda) parasitic in bark beetles Fadenwiirmer oder nematoden. I. Freile- in the United States. Proc. Helminthol. Soc. bende und pflanzenparasitische Nematoden. Wash. 24(1): 29-34. Tierwelt Deutschlands (Dahl) 36: 1-260. . 1958. Four new species of Parasity- SIDDIQI, M. R., AND J. B. GOODEY. 1963. The lenchus (Nematoda) from scolytid beetles. status of the genera and subfamilies of the Proc. Helminthol. Soc. Wash. 25(1): 26-30. Criconematiclae (Nematoda); with a comment . 1960. Nematode parasites and associ- on the position of Fergusobia. Nematologica ates of the California five-spined engraver, 9(3): 363-377. Ip* confusus (Lee.). Proc. Helminthol. Soc. SKARBILOVICH, T. S. 1947. Revision of the sys- Wash. 27(1): 14-22. tematics of the nematode family Anguillulin- 1962. Life history of Aphelenchulus ,idae Baylis and Daubney, 1926. Akademia elongatus Massey ( Nematoda), an enclopara- Nauk USSR 57(3): 307-308. site of Ips confusus LeConte, with a descrip- . 1959. On the structure of systematics of tion of the male. Jour. Insect Pathol. 4(1): nematodes, order Tylenchida Thorne, 1949. 95-103. MICOLETZKY, H. 1922. Die freilebenclen Erd- Acta Parasitol. Polonica 7(4): 117-132. Nematoden. Arch. Naturg. Abt. A. 87: 1-650. STRASSEN, OTTO K. L. ZUR. 1892. Bradijnema NICKLE, W. R. 1963a. Notes on the genus rigidiim v. Sieb. Ztschr. f. wissensch. Zool. Contortylenchus Riihm, 1956, with observa- 54(4): 655-747. tions on the biology and life history of C. THORNE, G. 1935. Nemic parasites and asso- elongatus (Massey, 1960) n. comb., a para- ciates of the mountain pine beetle (Dendroc- site of a bark beetle. Proc. Helminthol. Soc. tonus monticolae) in Utah. Jour. Agr. Res. Wash. 30(2): 218-223. 51(2): 131-144.

WACHEK, F. 1955. Die entoparasitischen Tylen- clrosophilid flies. Parasitology 49(1/2): 83- chiden. Parasitolog. Schriftenreihe, 3: 1—119 103. Jena, Fischer. WULKER, G. .1923. Uber Fortpflanzung und En- WARREN, E. 1941. On the occurrence of nema- twicklung von Allantonema und verwandten todes in the haemocoel of certain gamasid Nematoden. Ergebn. Fortschr. d. Zool. 5: mites. Ann. Natal Mus., 10(1): 79-94. 389-507. WELCH, H. E. 1959. Taxonomy, life cycle, de- . 1929. Bemerkungen zur Arbeit von G. velopment, and habits of two new species of Fuchs: Die Parasiten einiger Riissel-und Bork- Allantonematidae (Nematoda) parasitic in enkiifer. Z. Parasitenkcle 2(2): 286-290.

Encholaimoidea (Nematoda: Dorylaimida), A New Superfamily Representing Dorylaimid Specimens with Cephalic Setae

A. MORGAN GOLDENTI AND D. G. MuRPHY1

In the fall of 1963 nematodes obtained from as in other dorylaims. It is of the masculine a soil sample around the roots of coconut gender and is the latinized combination formed (Cocos nucifera L.) in Trinidad were sub- from the Greek "enchos," meaning spear, and mitted for identification by Dr. Karl Mara- "laimos," meaning throat. The specific Latin morosch, Boyce Thompson Institute for Plant name "taunts" means "bull," and refers here Research, Yonkers, New York. Among the to the presence of horn-like cephalic setae. nematodes found in this and other samples collected shortly thereafter from the same area Order Dorylaimida (De Man, 1876) were a total of about 30 specimens of a very Pearse, 1942 emended remarkable nematode. The amphids, spear, DIAGNOSIS: Nematoda, Adenophorea. Setae esophagus, and certain other structures were usually absent. Cuticle generally smooth; but clearly dorylaimid. Furthermore, the develop- sometimes heavily annulated, in some cases ment of the spear as seen in immature specimens, is the same as in other species of spear- longitudinal striae forming plate-like cuticle. bearing dorylaims. Yet, these specimens pos- Esophagus cylindrical, in 1 or 2 parts; when sess four small, and six prominent horn-like in 2, the posterior part conoid cylindrical or cephalic setae previously unknown in the pyriform. Glands uninucleate, and 3, 5, or 7 Dorylaimida. Also, the heavily annulated cuti- in number contained within the confines of cle divided into plates by numerous longi- the esophagus; all gland openings posterior to tudinal striae is unique in the dorylaims. the nerve ring. Head with 6—18 inner labial This interesting nematode is described here- papillae and 6, 10, or 14 outer papillae or in as the representative of a new genus, family, setae. Amphids cyathiform, pouch-like or tu- and superfamily. The Order Dorylaimida is bular. Stoma with mural tooth or teeth; with emended as specified below, so that this an axial stylet, or vestigial, or unarmed. Pre- nematode could be placed in this major group anal supplements usually present. to which it apparently belongs. Encholaimoidea, new superfamily The new generic name, Encholaimus, sug- DIAGNOSIS: Dorylaimida, sub-order Dory- gests "spear in the throat," referring to the laimina. development of the spear in the esophagus Head with setae. Stoma armed. Amphids pouch-like, with slit-like apertures. Esoph- 1 Nematologists, Nematology Investigations, Crops Re- agus composed of a slender anterior part search Division, Agricultural Research Service, U. S. De- partment of Agriculture, Beltsville, Maryland. and an expanded posterior portion. Excretory

Fig. 1. Full length drawing of female of Encholaimus taunts, n. gen., n. sp.

Copyright © 2011, The Helminthological Society of Washington 96 PROCEEDINGS OF THE [VOL. 34, No. 1 pore absent. No caudal glands. Supplements pecially). Amphid apertures large, slit-like. present. Cuticle heavily annulated, with longitudinal The presence of cephalic setae is a distin- striae, numbering 24 at mid-body, forming guishing feature of this dorylaimid superfamily. a plate-like appearance. Spear delicate, with distinct guide ring; spear extension swollen Encholaimidae, new family posteriorly, with large flanges as illustrated. DIAGNOSIS: Encholaimoidea. Cuticle heavily Esophagus tubular for much of its length, annulated, often with longitudinal striae giving terminating posteriorly in a pyriform, basal a plate-like appearance. Stoma armed with an bulb measuring about 15% of the esophageal axial spear. Anterior portion of esophagus tu- length. Intestine filled with globular granules, bular, posterior third or less enlarged, without details obscure. Ovary 1, dorylaimid, extend- a spiral sheath. ing posteriorly and then reflexed for almost The type of cuticle as described, plus the one-half its length. Vulva a transverse slit, presence of cephalic setae, distinguish this located in anterior half of body. Anus distinct. from other families of the Dorylaimicla. Tail conical, appearing as in Fig. 1, averaging TYPE GENUS: Encholaimus, n. gen. 39 p.. Lateral pores not detected. ALLOTYPE (male*): Length 0.56 mm; a = Encholaimus, n. gen. 29; b — 4.5; c — 18; spear 18 ^, spear extension 13; total 31 fj,. DIAGNOSIS: Encholaimidae. Body of both Male similar to female in general size and sexes cylindroid, tapering at extremities. Head shape. Body width 19 p.. Head, cephalic setae, with 6 large, horn-like and 4 smaller setae in cuticle, spear and spear extensions, and esoph- outer circle. Six labial or perioral papillae in agus also as in female. Testes 2, details ob- inner circle. Spear with swollen extensions, scured. Spicules paired, dorylaimid in shape, bearing knobs or flanges. Cuticle heavily an- measuring 28 p.. Gubemaculum absent. Tail nulated, with longitudinal striae forming a curving ventrally, conical in shape as illustrated plate-like pattern. Lateral pores absent. Ovary in Fig. 2 E, and measuring 31 ^ in length. 1; vulva a transverse slit. Spicules paired, Two preanal supplements present. dorylaimoid in shape. DIAGNOSIS: Encholaimus with above mea- Distinctive on the basis of the generic char- surements and description. No other species acters as specified. No other known genus. known. HOLOTYPE (female): Collected by Karl TYPE SPECIES: Encholaimus tannis, n. sp. Maramorosch November, 1963 and received Encholaimus taurus, n. sp. 20 November 1963. Trinidad. Slide T-83t, (Figs, land 2 A-E) United States Department of Agriculture Nem- MEASUREMENTS (15 females): Length 0.55 atode Collection, Beltsville, Maryland. mm (0.47-0.61); a = 27.1 (24-29); b = 4.8 ALLOTYPE (male): Same data as holotype. (4.1-5.5); c = 13.8 (13-15); V = 40% (37- Slide T-84t, United States Department of 41); spear 18 ^ (17-20); spear extensions Agriculture Nematocle Collection, Beltsville, 12 fj, (11-12); total 30 p (29-31). Maryland. HOLOTYPE (female): Length 0.56 mm; a = PARATYPES (females and larvae): United 27; b = 4.9; c = 14; V = 40%; spear and States Department of Agriculture Nematode spear extensions 31 //,; body width 21 ^ tail Collection, Beltsville, Maryland; and two fe- 39 ^ in length. males, California Nematode Survey Collection, DESCRIPTION OF FEMALES: Body tapering Davis, California. anteriorly and more so posteriorly as illustrated TYPE HABITAT, HOST AND LOCALITY: Soil (Fig. 1). Body width averaging 20 jM,. Head around roots of coconut (Cocos nucifcra L.) with a prominent outer circle of 10 horn-like at Golden Grove, Trinidad. cephalic setae, 6 of which are large while 4 DISCUSSION: The nematode described above are about one-half the size. These surround an inner circle of 6 conspicuous labial papillae * Only one male has been found, and unfortunately it was almost filled with an unidentified sporozoan parasite, on a labial crown (see Fig. 2 A and D es- obscuring internal details.

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Fig. 2. Drawings of Encholaimiifs taurus, n. gen., n. sp. A, Anterior portion of female. B, Anterior part of immature molting specimen. C, Cross-section of female at mid-body showing, particulally, the cuticle. D, En face view of female. E, Posterior portion of male.

Copyright © 2011, The Helminthological Society of Washington 98 PROCEEDINGS OF THE [VOL. 34, No. 1 obviously has a relationship to other dorylaims, setae and the heavily annulated, plate-like particularly some forms in the Leptonchidae. cuticle, have not been previously known in The esophagus, spear and extensions, repro- the Dorylaimicla. The cephalic setae in par- ductive structures, and development of the ticular suggest a relationship to the free-living spear in the esophagus of molting specimens marine forms of the Enoplida. Further studies all are characters of the dorylaimid type. On on this, and related forms, will be required to the other hand, the two very outstanding fea- establish more clearly the phylogenetic rela- tures of this nematode, the prominent cephalic tionships in the dorylaims.

Biology of Mastophorus nuniidica (Seurat, 1914) Read and Millemann, 1953 (Nematoda: Spiruridae) with a Description of the Juvenile Stages1

WILLIAM G. DYER2 AND O. WILFORD OLSEN

Life histories of several species of Masto- granella) could serve as intermediate hosts of phorus have been investigated. Leuckart M. muris. (1867) and Marchi (1871) showed that M. Brumpt (1931) found that M. bonnet (Ort- muris (Gmelin, 1790) Chitwood, 1938 de- lepp, 1924) Read and Millemann, 1953 from velops in meal worms (Tenebrio molitor). domestic rats develops in cockroaches (Rhy- Adults were obtained in Mus decumanus parobia maderae, Blatella germanica, and Peri- which had eaten infected meal worms. Cram plan-eta oriental^). M. muricola (Gedoelst, (1926) recovered encysted juveniles from the 1916) Read and Millemann, 1953 was reported body cavities of cockroaches (Blatella ger- from three species of captive monkeys (Cebus manica) fed embryonated eggs of M. muris capucinus, Ateles dariensis, and Aotus zonalis ( — Protospirum columbiana Cram, 1926). by Foster and Johnson (1939). Infective juve- Adults were obtained experimentally in rats. niles were identified from the body cavities of Hall (1929) showed that scarabaeid beetles cockroaches (Leucophaea maderae). (Aphodius fimetarius) are natural intermediate Crook and Grundmann (1964) exposed 15 hosts of M. muris (= Protospirura gracilis species of native insects occurring naturally Cram, 1924). Marcandier and Pirot (1937) in association with deer mice and four com- observed juveniles of M. muris in the oriental mon laboratory insects to eggs of M. numidica. rat flea (Xenopsylla cheopis} collected from Only tenebrionid beetles (Eleodes tuberculata Mus decumanus. Adults were found in the patruelis) were naturally and experimentally stomach of rats. Miyata (1939) demonstrated infected. that cockroaches (Periplaneta americana), Because of the need for more detailed infor- fleas (Leptopsi/lla musculi, Ceratophyllus ani- mation on the life cycle in Mastophorus, fur- sus, C. fasciatus, X. cheopis), and moths (Tinea ther study of M. numidica. was undertaken. On the basis of the findings, an account of

1 From the Department of Zoology, Colorado State Uni- the life cycle and descriptions of the morphol- versity, Fort Collins, Colorado. ogy of the developmental stages are presented A portion of a dissertation submitted to the Graduate School, Colorado State University, in partial fulfillment of in this paper. the requirements for the degree of Doctor of Philosophy. This investigation was supported hy a training grant (5 Tl Al 94-05) from the Institute of Allergy and Infec- tious Diseases of the National Institutes of Health, U. S. MATERIALS AND METHODS Public Health Service. 2 Present address: Department of Biology, Minot State Infective eggs containing first-stage juveniles College, Minot, North Dakota. Appreciation is expressed to Dr. Albert W. Grundmann, were obtained as required by immersing ovig- Head of the Department of Zoology and Entomology, Uni- erous female worms from the stomachs of versity of Utah, for confirmation of specimens of M. numidica and valuable suggestions. P. maniculatus in physiological saline. A single

Copyright © 2011, The Helminthological Society of Washington JANUARY, 1967] HELMINTHOLOGICAL SOCIETY 99 feeding of eggs was given to individual grass- TABLE 1. Results of feeding 40 eggs of Masto- hoppers (Melanoplus femur-rubrum), crickets phorus numidica per insect to grasshoppers (Me- (Acheta domestica), and beetles (Eleodes ob- lanoplus femur-rubrum), crickets (Acheta domestica), and beetles (Eleodes obsoleta); 24 days after soleta) which had been starved for at least exposure. one day. Specimens of each species were vised as controls to determine if natural infections No. of cysts No. No. recovered Insects examined infected were present. All test and control specimens Mean Range of M. femur-rubrum and E. obsoleta were col- M. femur-rubrum lected in the Cache la Poudre Canyon, Lari- Exposed 50 31 9.5 2-19 Control 42 1 — — mer County, northern Colorado. Specimens of A. domestica A. domestica were obtained from Baton Rouge, Exposed 46 18 8.3 6-18 Control 95 0 — — Louisiana. Anesthetized insects were pinned E. obsoleta Exposed 20 18 8.9 1-15 to the bottom of plastic petri dishes, dissected Control 12 4 0.5 1-2 in 87.6 per cent Ringer's solution, and examined for the presence of juveniles. Deer mice used for experimental infections RESULTS were determined to be free of M. numidica by frequent examination of the feces over a period Experimental infections were obtained in all three species of insects (M. femur-rubrum, of several months. Some were fed infected A. domestica, and E. obsoleta) fed eggs of insects and others isolated cysts containing third-stage juveniles removed from the hemo- M. numidica (Table 1). Control specimens coels of insects. The feces of these deer mice of M. femur-rubrum and E. obsoleta showed natural infections of 2.4 and 33.3 per cent, were checked daily for eggs to determine the respectively. prepatent period. Following is an account of the life cycle Infected mice were examined by opening of M. numidica as it occurs in the cricket (A. the esophagus, stomach, and small intestine domestica) and the deer mouse. The verbal separately and agitating each part in a jar descriptions and the illustrations are based on of saline to free the worms, the majority of specimens from these hosts. which was found readily upon gross examination. The mucosa was scraped from the walls Development and morphology of each part of the alimentary canal and placed in the cricket in separate containers of warm saline to allow About 3.5 to 4.5 hr postinfection, the crop embedded juveniles to migrate from the tissues contained eggs which were 0.054 to 0.059 by into the fluid. The contents of the containers 0.043 to 0.046, embryonated, thick shelled, were stirred, sedimented, decanted several and otherwise indistinguishable from those oc- times, and examined for juveniles after addition curring in the definitive host's feces. At about of fresh saline. 7.5 hr most eggs were in the stomach, a few Microscopic preparations were made of both still in the crop and gizzard; some evidently living and fixed worms. Since certain struc- had hatched, as first-stage juveniles were found tures appeared more clearly in living speci- in the intestine. At about 10 to 15 hr neither mens, some juveniles were relaxed in saline by eggs nor juveniles were observed in the diges- gently applying heat sufficient to reduce active tract. tivity. Others were killed in hot saline, fixed FIRST-STAGE JUVENILE: At 10.5 hr post- in 70 per cent ethanol and 3 per cent gly- exposure, the abdominal portion of the hemo- cerine, cleared, dehydrated, and mounted in coel contained a few first-stage juveniles (Fig. glycerine. A phase contrast microscope was 1). Five days after exposure both free and used in morphological studies. encysted first-stage juveniles were present, and Drawings of the several stages were pre- measured 0.392 to 0.407 by 0.045 to 0.055; pared from fixed and living specimens. All anterior end rounded; posterior portion taper- measurements are in millimeters except where ing slightly with tip of tail ending in character- otherwise indicated. istic short conical process; cuticle thin, trans-

TABLE 2. Age at which third-stage juveniles of TABLE 3. Experimental infection of deer mice by Mastophorus numidica become infective in cysts feeding each a single cricket exposed to eggs of in Acheta domestica. Mastophorus numidica. No. of Age of Approximate No. of juveniles No. of No. of eggs Time after No. of juveniles deer cyst No. of cysts recovered/ deer fed to exposure recovered/ mice ( days ) given/mouse mouse mice crickets (days) mouse 2 10 35 0 2 40 10 0 3 22 20 0 3 40 22 0 5 27 20 0 3 40 27 0 4 32 20 0 4 50 32 0 1 35 30 0 3 50 35 0 3 38 30 10 3 50 38 12 3 40 30 13 3 50 40 15 2 45 35 7 2 50 45 13 parent, with very fine transverse striations; 0.130 to 0.135 from anterior extremity, respec- oral opening leading into a transparent esoph- tively. agus approximately one-fifth of body, slightly Juveniles recovered on the 19th day post- swollen at posterior end; intestine granular, infection were molting. The partially detached connecting posteriorly with a very short rectum cuticle was observed at the posterior end (Figs. surrounded by two rectal glands (Fig. 2). 5, 6). By the 21st day, a few juveniles still Juveniles undergo the first molt 7 to 9 days showed a partially detached cuticle while the after experimental infection as indicated by majority had completed the second molt and a detached cuticle in the tail region (Fig. 3). were in the third stage of development. On the 8th day, some showed a partially de- THIRD-STAGE JUVENILE: By the 22nd day, tached cuticle, others had completed the first third-stage juveniles only were observed. Those molt and were in the second stage. recovered on the 24th day measured 2.684 to SECOND-STAGE JUVENILE: Only second-stage 2.737 by 0.094 to 0.099; body tapering toward juveniles were observed by the ninth day. The each extremity; tail conical terminating in a bulbous swelling at the posterior part of the characteristic rosette of spinous processes (Fig. esophagus was not as prominent as in the pre- 10); cuticle thick and transversely striated; ceding stage. Fourteen-day-old juveniles mea- lips trilobed similar to adult, median lobe larger sured 2.068 to 2.189 by 0.77 to 0.81; body than dorsal or ventral (Fig. 7), each lobe tapering toward each extremity; tip of tail armed with two teeth; four submedian papillae rounded, having lost the conical process with present, located at bases of dorsal and ventral shedding of first cuticle (Fig. 4); cuticle thin, lobes; buccal capsule 0.040 to 0.042 deep; transparent and with very fine transverse stria- esophagus similar to that of second-stage juve- tions; oral opening leading into a buccal cap- nile but broader near posterior end; nerve sule 0.001 to 0.012 deep; esophagus about one- ring and excretory pore 0.125 to 0.128 and third of body length, divided into muscular and 0.145 to 0.148 from anterior end, respectively. glandular portions, terminal swelling absent; Sex could be differentiated at this stage; male intestine simple and narrow; rectum sur- genital primordium elliptical in shape about rounded by three large rectal glands; nerve 0.032 to 0.034 by 0.015, located ventrally ring and excretory pore 0.108 to 0.111 and between body wall and intestine, 0.630 to

Figs. 1-13. Various stages in the development of Mastophorus numidica, drawn with the aid of camera lucida. 1-2. First-stage juvenile, complete worm. 3. First-stage juvenile, undergoing first molt, caudal end. 4. Second-stage juvenile, posterior extremity, lateral aspect. 5-6. Second-stage juvenile, undergoing second molt, lateral aspect of caudal end. 7. Third-stage juvenile, anterior extremity, lateral aspect. 8. Genital primordium attached to body wall, third-stage juvenile female, lateral view. 9. Genital primordium of third-stage juvenile male, lateral view. 10. Third-stage juvenile, posterior extremity, lateral aspect. 11. Third-stage juvenile, prior to third molt, lateral aspect of caudal end. 12. Third-stage juvenile, undergoing third molt, lateral aspect of caudal end. 13. Fourth-stage juvenile, undergoing fourth molt, dorsal aspect of caudal end.

TABLE 4. Determination of the prepatent period stage females only; body 7.425 to 8.360 by of Mastophorus numidica in the deer mouse. 0.187 to 0.196; cuticle thick and transversely No. of Age of striated; lips well developed; buccal capsule No. of encysted cysts Ova first deer juveniles from appeared 0.072 to 0.080 deep; nerve ring and excretory mice administered/ crickets in feces ( days ) pore 0.180 to 0.192 and 0.252 to 0.268 from mouse (days) anterior end, respectively; primitive vulva pos- 1 25 10 terior to mid-body; vagina short and curved 1 25 22 -••• 1 23 27 '•':'- posteriorly; muscular ovijector followed by 2 24 32 '•'.'• 4 25 35 -:' short trunk which branches into an anterior 3 22 38 35 and a posterior uterus; oviduct narrow; tail 35 conoid with small cuticular elevations at tip 41 4 22 40 37 giving it a rough appearance. 40 42 Two females recovered from the stomach of 42 a deer mouse examined 15 days postinfection 2 22 45 35 37 were beginning the fourth molt. They showed * Ova absent in one daily examination. a partially detached thick outer cuticle with cuticular elevations at the caudal tip and the thick inner cuticle was smooth at the tip of 0.635 from posterior end, composed of 2 large the tail (Fig. 13). By the 17th day, all speci- epithelial cells enclosing a group of germinal mens examined had completed the fourth molt cells (Fig. 9); female genital primordium also and were fifth stage. The adult stage has been somewhat elliptical, about 0.031 by 0.011, described by Seurat (1914) and warrants no attached to body wall ventrally by means of a further description here. cell, 0.874 to 0.876 from tip of tail (Fig. 8). Determination of prepatent period Development and morphology of ivorms in the deer mouse At 35 to 42 days after ingestion of infective juveniles, sexually mature adults were present Infection was established in deer mice fed as evidenced by the appearance of eggs in the 38- to 45-day-old encysted juveniles removed feces of experimentally infected clecr mice from the hemocoels of crickets (Table 2) as (Table 4). well as in those fed crickets 38 to 45 days after exposure of the latter to eggs of M. DISCUSSION numidica (Table 3). Similar results were ob- With the exception of studies reported by tained with grasshoppers and beetles. Since Hall (1929) and Crook and Grundmann older juveniles were not administered, it was (1964), all prior life history investigations of not determined if infectivity of juveniles de- Mastophorus were conducted with insects creases with age. Transition to the fourth stage occurred in which are commonly used in the laboratory the stomach of the deer mouse 8 to 11 days and probably are not the intermediate hosts after ingestion of cysts containing infective under natural conditions. Though several third-stage juveniles. In a late phase of the orders of insects (Coleoptera, Orthoptera, impending molt, two thick cuticles are clearly Siphonaptera, and Lepicloptera) have been observable, an outer one with a rosette of found to contain members which serve as spinous processes at the terminus of the tail intermediate hosts of Mastophorus, grasshop- and an inner one with small elevations at the pers have not been reported previously to caudal tip (Figs. 11, 12). By the end of the function as such. Results with other orthop- llth day, all juveniles examined had com- teraii insects suggest that grasshoppers may pleted the third molt and were in the fourth play an important role under natural condi- stage. tions as intermediate hosts for species of FOURTH-STAGE JUVENILE: Examination of a Mastophorus other than M. numidica. deer mouse 14 days after ingestion of cysts Crook and Grundmann (1964) reported that containing third-stage juveniles revealed fourth- grasshoppers (M. femur-rubrum) did not be-

Copyright © 2011, The Helminthological Society of Washington JANUARY, 1967] HELMINTHOLOGICAL SOCIETY 103 eome infected when exposed to feces of deer from the hemocoel of crickets as well as in mice containing eggs of M. numidica. How- those fed crickets 38 to 45 days after exposure ever, in the present experiment, grasshoppers of the latter eggs of M. numidica. Similar became infected when fed the eggs concen- results were obtained with grasshoppers and trated on a small piece of lettuce which had beetles. The fourth and fifth molts occur in been thoroughly washed with tap water. Since the stomach of deer mice 8 to 11 and 15 to 17 grasshoppers are not coprophagic, it seems days postinfection, respectively. The prepatent probable that they become infected in nature period ranged from 35 to 42 days. by eating contaminated vegetation rather than LITERATURE CITED fecal matter. The time intervals required in the inter- BHUMPT, E. 1931. Nemathelminthes parasites mediate hosts for development to infectivity des rats sauvages (Epimijs norvegicw) de Caracas. I. Protospirura honnei. Infections by juveniles of M. muris and M. numidica are experimantales et spontanees. Formes adnltes similar. Cram (1926) found that 41 clays were et larvaires. Ann. Parasitol. 9: 344-358. necessary for development of infective M. CRAM, E. B. 1926. A new nematode from the muris in the hemocoel of Blatella germanica. rat and its life history. U. S. Nat. Mus., Proc. Crook and Grundmann (1964) reported that 68: 1-7. infective cysts occurred in the hemocoel of CROOK, J. R., AND A. W. GRUNDMANN. 1964. E. tuberculata patruelis 40 days after inges- The life history of Protospirura numidica tion of eggs of M. numidica. Similar results Seurat, 1914 (Nematoda: Spiruroidea). Proc. Helminth. Soc. Wash. 31: 225-229. were obtained in the present study; 38 days FOSTER, A. O., AND C. M. JOHNSON. 1939. A after ingestion of eggs of M. numidica by preliminary note on the identity, life cycle, the grasshopper (M. femur-rubrum), infective and pathogenicity of an important nematode cysts were observed in the hemocoel. The pre- parasite of captive monkeys. A. ]. Trop. patent period reported for M. muris is about Med. 19: 265-277. 115 days as compared with about 36 days for HALL, M. C. 1929. Arthropods as intermediate M. numidica, as found by Crook and Grund- hosts of helminths. Smithson. Misc. Collect. mann (1964). In the present study, this period (Publication 3024) 81: 77 p. LEUCKART, K. 1867. Die menschlichen Parasiten ranged from 35 to 42 days. und die von ihnen herriihrenden Krankheiten. Ein Hand-und Lehrbuch fur Naturforscherund SUMMARY Aerzte. Vol. 2 Leipzig U. Heidelberg. 256 p. An account of the life cycle of Mastophorus MARCANDIER, M., AND R. PIROT. 1937. Etude numidica as it occurs in the cricket (Acheta sur les ectoparasites des rats de Toulon. Bull. domestica} and the deer mouse (Peromijscus Soc. Path. Exot. 25: 237-244. maniculatiis) is described and illustrated. Ex- MARCI-II, P. 1871. Monografia sulla storia genet- ics e sulla anatomia della Spiroptera obtusa perimental infections were obtained in all three Rud. Mem. R. Aclad. Sc. Torino, Cl. Sc. Fis. species of insects (Melanoplus femur-rubrum, e Mat. 25: 1-30. Eleodes obsoleta, and A. domestica) fed eggs MIYATA, I. 1939. Studies on the life history of of M. numidica. Control specimens of M. the nematode Protospirura muris (Gmelin) femur-rubrum and E. obsoleta showed natural parasitic in the stomach of the rat, especially infections. The first and second molts occur on the relation of the intermediate hosts, in the hemocoel of crickets 7 to 9 and 19 cockroaches, skin moth, and rat fleas. Vol. Jub. Yoshicla 1: 101-136. to 22 days postinfection, respectively. SEURAT, L. B. 1914. Sur un nouveau spiropetre Infection was established in deer mice fed clu chat grante. Compt. Rend. Soc. Biol., 38- to 45-day-olcl encysted juveniles removed Paris 17: 344-347.

Annotated Record of Some Previously Described Digenetic Trematodes of Amphibians and Reptiles from the Philippines, Korea, and Matsu Island1

JACOB II. FlSCHTHAL AND ROBERT E. KuNTZ"

The trematodes of this report were part of MEASUREMENTS AND SOME PERTINENT DATA a collection made by the junior author while (based on eight adults and 40 immature worms a member of the U. S. Naval Medical Research from nine hosts; one mature and two young Unit No. 2, Taipei, Taiwan. While this report adults measured): Body 1,645 to 2,475 by contains no new forms we are presenting the 270 to 780; preoral body 0 to 16; pigment data in order to supplement previous body and granules of disintegrated cercarial eyespots organ measurements so that worms of different concentrated from just postbifurcal to posterior size ranges, which may represent differences half of oral sucker but may extend beyond due to allometric growth, are not described as these levels; oral sucker 136 to 250 by 111 new species; to note intraspecific variations to 138; esophagus 189 to 295 long; acetabulum so that such variants are not described as new 370 to 642 by 410 to 575; testis 157 to 240 species; to detail anatomical descriptions which by 121 to 295; cirrus sac 60 to 110 by 48 to have been lacking or which have been pre- 110; ovary 77 to 194 by 61 to 194; vitellaria viously presented without such detail; to gather confluent anteriorly and usually posteriorly, additional data for clarifying statements in the interrupted at testis level, 21 to 25 follicles literature which may be questionable; to indi- anterior to interruption and 16 to 19 posterior; cate new and probable synonymies; and to cecal bifurcation to genital pore 205 to 420, note new host and geographic distribution to testis 370 to 420, to ovary 595 to 835; nine records. Parasites were washed in saline, killed eggs 96 to 107 by 66 to 80. in hot water, and transferred immediately to DISCUSSION: This species was originally de- FAA fixative. After 4-8 hours they were scribed by Tubangui (1933) from Rana spp. stored in 70% alcohol plus 2% glycerine. All from the same island as the present specimens; measurements are in microns. later, Tubangui (1947) specifically identified Rana vittigera as the only host. Li (1937a) FAMILY PARAMPHISTOMIDAE described a new species, D. sinicus, from the Diplodiscus amphichrus Tubangui, 1933 same host species as our specimens from China and from R. regulosa Wiegmann from China SYNONYM: Diplodiscus sinicus Li, 1937. and Amoy Island. Bravo (1941) declared Li's HOST: Rana limnocharis vittigera (Giinther) species a synonym of D. amphichrus. Our (syn. R. vittigera Wiegmann) (Ranidae). nine hosts harbored one (mature adult), two HABITAT: Small intestine. (two mature adults in one, two immature in LOCALITY: Manila, Luzon Island, Philip- another), three (one young adult and two pines. immature in one, three immature in each of DATES: 11, 13 December 1961. two others), seven (immature), nine (three SPECIMENS: USNM Helm. Coll. No. 61700 young adults and six immature), and 18 (one (three slides with one adult specimen each). mature adult and 17 immature) worms, re-

L Contribution from the Department of Biology, State spectively. University of New York at Binghamton (J. H. Fischthal). - Address of R. E. Kuntz: Southwest Foundation for Research and Education, San Antonio, Texas. FAMILY PLEUROGENIDAE This work supported by funding under Public Law 480, Section 104(c), through funds provided by the Bureau of Pleura genoides taylori (Tubangui, 1928) Medicine and Surgery, Navy Department Work Unit MF 022.03.07-2003 and with aid provided by Contract ONT Travassos, 1930 (NR 103-690/N0014-66-C0094) between the Office of Naval Research and Southwest Foundation for Research SYNONYMS: Pleurogenes taylori Tubangui, and Education. The opinions and assertions herein are those of the authors and are not to be construed as official 1928; Pleurogenoides hashimi Rohde, 1963. or reflecting the views of the Navy Department or the Naval service at large. The authors are indebted to Dr. HOST: Rana limnocharis vittigera (syn. R. Robert F. Inger, Curator of Reptiles, Chicago Natural History Museum, for host identifications. vittigera) (Ranidae).

TABLE 1. Pleurogenoides taylori. Measurements of eight specimens in microns. Specimen No. 1 2 3 4 5 6 7 8 Body 284X133 286X121 302X143 370X174 495x265 506X318 705X475 980X501 Forehocly 145 160 167 — 290 264 276 342 Hindbody 82 70 77 — 108 133 245 285 Oral sucker 59X61 69X69 67X68 73X78 97X102 85X99 138X153 133X155 Acetabulum 57X57 56X52 58X56 — 97x97 109X90 132X132 136X143 Pharynx 28X34 34X33 28X34 31X42 51X49 48x57 — 76X57 Right testis 85X46 55X44 77X60 99X73 85X73 169x148 145x138 157X172 Left testis 93X40 53X46 77X59 94X51 126X70 128X108 145X134 184X111 Cirrus sac 160X59 183X34 131X44 150X46 281X85 283X70 290X136 345X159 Ovary — 46X46 44x34 46X38 85X85 99x119 133x99 — — — — Seminal receptacle — — — 77X97 65X80 Eggs — — — — — 30-31X15-16 28-32X15 30-32X15-18

HABITATS: Stomach, small intestine. b — constant. He converted this formula into LOCALITY: Manila, Luzon Island, Philip- log y = log b + k log x which corresponds pines. to a straight line when shown graphically in DATE: 13 December 1961. a double-logarithmic system of coordinates. SPECIMENS: USNM Helm. Coll. No. 61701 Rohde (1966) applied this method in deter- (three slides with one adult worm each). mining species synonymy in the genus Anchi- MEASUREMENTS AND SOME PERTINENT DATA trema Looss, 1899 (Lecithodendriidae). We (based on two mature adults from one host, have followed Rohde's methods in attempting and two young adults and six immature from to show further that P. hashimi is a synonym a second): Measurements for all but two im- of P. taylori. Our graphs substantiated our mature specimens presented in Table 1. Sucker opinion. However, to ascertain whether we length ratio 1:0.81 to 1.28; in four adults pre- were justified in interpreting the graphs as oral body 3 to 10 long and vitelline follicles we did we similarly plotted on the same graphs 14 to 16 in number. most of the other species of Plewogenoides DISCUSSION: In Table 1 specimens 1 to 4 Travassos, 1921 (Figs. 1 to 6). The graphs are immature, 5 has vitelline cells and shell show interestingly that most species of the material in the uterus but no eggs, 6 is a young genus, with the exception of P. bufonis Kaw, adult with eggs, and 7 and 8 are mature adults. 1943 and P. minus (Pigelevsky, 1931), exhibit This species was described by Tubangui (1928) basic similarities in allometric growth of the from the intestine of Rana vittigera from Luzon body and its organs. We can not conclude and redescribed by Li (1937a) from R. regu- that all but the two exceptions are synonymous losa from China. with P. taylori; for example, P. solus (S. J. We declare P. hashimi, described by Rohcle Johnston, 1912) has significantly smaller eggs (1963) from Rana cancrivora Gravenhorst (20 microns long) and the number, size, and from Malaya, a synonym of P. taylori. Rohde, distribution of the vitelline follicles are quite without considering Li's description of P. different for P. compactiis Shtrom, 1940 and taylori, stated that his form appears closest P. gastropprus (Liihe, 1901). It may be that to the latter species but differs in its larger several of the remaining species are synon- size, distinctly larger testes and ovary, and ymous but further study is necessary. There smaller eggs '(28 to 33 by 11 to 13). The is a danger in overemphasizing allometric latter point is invalid as Tubangui gave the growth in determining species synonymy since egg size as 31 to 33 by 15 to 18 and Li as many species within any single genus may 27.3 to 31.2 by 11.7 to 15.6. There is an show much similarity. Fortunately, Rohde overlap in gonad size with Li's specimens. (1966) had in his collection specimens of The relative differences in body and gonad Anchitrema covering the size ranges of the sizes may be due to allometric growth as all species he placed in synonymy. described specimens of P. taylori, as well as FAMILY PLAGIORCHIIDAE ours, are smaller than those of P. hashimi. Huxley (1932) expressed allometric growth Glypthelmins staffordi Tubangui, 1928 by the formula y = bxk, where y = organ size, HOST: Rana limrwcharis vittigera (syn. R. x — body size, k = allometric exponent, and vittigera) (Ranidae).

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HABITATS: Stomach, small intestine. cirrus sac; 32 operculate eggs 29 to 37 by 19 LOCALITY: Manila, Luzon Island, Philip- to 27. pines. DISCUSSION: The eight infected hosts har- DATES: 11, 13 December 1961. bored one (adult), two (adult, in two), three SPECIMENS: USNM Helm. Coll. No. 61702 (adult), four (one adult and three immature (four slides with one adult each). in one, four immature in another), seven (four MEASUREMENTS AND SOME PERTINENT DATA adult and three immature), and 21 (11 adult (based on 24 adult and 20 immature worms, and 10 immature) worms, respectively. This eight adults measured): Body 1,454 to 1,960 species was described by Tubangui (1928) by 397 to 625; forebody 465 to 645, hindbody from Rana vittigera from Luzon, and rede- 780 to 1,286, preoral body 12 to 39, posttes- scribed by Li (1937a, b) from Rana regulosa ticular space 615 to 1,015, postvitellarian space Wiegmann from China, by Yamaguti and 320 to 475, postcecal space 120 to 440; oral Mitunaga (1943) from Bufo melanostictus sucker 151 to 206 by 152 to 222; acetabulum Schneider (Bufonidae) from Formosa and by 80 to 125 by 96 to 130, center at level of Yuen (1962) from Rana macrodon Kulil, R. anterior one-third to four-tenths of body length; erythraea (Schlegel) and R. cancrivora Graven- sucker length ratio 1:0.48 to 0.67; prepharynx horst from Singapore. Nasir (1966) briefly 15 to 39 long; pharynx 75 to 111 by 75 to 121, reviewed the genus. transversely or longitudinally elongate; esophagus 10 to 85 long; cecal bifurcation 111 to 184 Styphlodora renalis Tubangui, 1933 preacetabular; testes round to longitudinally or HOST: Naja naja (L.) (Elapidae). transversely elongate; left testis 157 to 206 by HABITATS: Kidney, small intestine. 160 to 206, usually more anterior than right LOCALITY: Zamboanga, Mindanao Island, testis, overlapping acetabulum as much as 105 Philippines. to lying entirely postacetabular as much as 44; DATES: 23 January, 16 February 1961. right testis 162 to 215 by 114 to 189, overlap- SPECIMENS: USNM Helm. Coll. No. 61703 ping acetabulum as much as 57 to lying en- (eight slides with two specimens each from tirely postacetabular as much as 92; cirrus sac kidney and one slide with one worm from small 99 to 182 (longitudinal extent) by 63 to 87, intestine). wall moderately thick, muscular, anteriormost MEASUREMENTS AND SOME PERTINENT DATA margin 41 to 92 preacetabular; ovary 111 to (based on 63 adults from kidney of one cobra, 143 by 114 to 186, smaller than testes but 11 measured): Body 1,657 to 2,827 by 560 occasionally may be slightly wider, round to to 1,210; forebody 445 to 550, hindbody 965 longitudinally or transversely elongate; vitel- to 2,000, posttesticular space 472 to 1,080, laria may commence at level about halfway postcecal space 200 to 570; suckers round to between cecal bifurcation and acetabulum, that longitudinally or transversely elongate; oral is, slightly anterior to anteriormost margin of sucker 157 to 227 by 167 to 230; acetabulum

EXPLANATION" OF GRAPHS Key to symbols in Figures 1—6: O Pleurogenoides tai/lori (Tubangui, 1928); aver- P. gastroporus, after Simha, 1958; average data. age data. V P.. hashimiMu.vumt RohdeixuiiLic,, 1963-Lii/u-j, ; 11111individua1 viiiuell l uuuidata. . © P. taylori, after Li, 1937: average data. X P'.. ifranensis DollfusDollfi'L , 1C11958^«-; iv.rl;,,,vl,,oindividual ,!.,data< . • P. taylori, our specimens; individual data. P. japonicus (Yamaguti, 1936); average data. 0 P. bufonis Kaw, 1943; average data. P. minus (Pigelevsky, 1931); average data. A P. compactus Shtrom, 1940; average data. P. sitapurii (Srivastava, 1934); average data. A P. freycineti (S. J. Johnston, 1912); average data. P. sphaericus (Klein, 1905); average data. [s] P. gastroporus (Liihe, 1901); average data. p. solus (S. J. Johnston, 1912); average data. • P. gastroporus, after Travassos, 1930; average '. tenet (Looss, 1898), after Travassos, 1930; data. average data. The size of the specimens of Pleuro^enoides and their organs is given as average diameter calculated from two dimensional measurements: (length -f- width)/2.

215 to 325 by 220 to 360, center at level of 255 to 720 long, left 189 to 515; right field approximate anterior one-fourth to one-third of extending 155 to 540 postacetabular, left 132 body length; sucker length ratio 1:1.11 to to 485; 42 operculate eggs 36 to 51 by 18 1.47; prepharynx 17 to 49 long; pharynx 97 to 25. to 136 by 84 to 114, longer than wide by 9 MEASUREMENTS AND SOME PERTINENT DATA to 31; esophagus 24 to 63 long; cecal bifurca- (based on one adult from small intestine of tion close to acetabulum or well separated, 48 second cobra): Body 2,812 by 595; forebocly to 182 preacetabular; testes round to longi- 627, hindbody 1,950, posttesticular space tudinally or transversely elongate, smooth to 1,100, postcecal space 177; oral sucker 196 slightly irregular or somewhat lobed, usually by 193; acetabulum 235 by 230, center at relatively narrowly separated from one another level of anterior one-fourth of body length; by ascending uterus but may be widely sep- sucker length ratio 1:1.19; prepharynx 52 long; arated or in contact, usually diagonal in posi- pharynx 120 by 128; esophagus 74 long; cecal tion with left testis anterior to right but almost bifurcation well separated from acetabulum, symmetrical in some; anterior testis 197 to 177 preacetabular; testes smooth; anterior testis 330 by 177 to 350, 35 to 275 postacetabular; 260 by 230, 342 postacetabular; posterior testis posterior testis 206 to 330 by 186 to 400, 197 395 by 240, 455 postacetabular; cirrus sac 475 to 625 postacetabular; cirrus sac 265 to 605 (longitudinal extent) by 113, commencing 134 (longitudinal extent) by 90 to 152, very thick postacetabular, anteriormost extent 111 pre- walled, muscular, proximal portion may be acetabular; seminal vesicle long, tubular, bi- straight or bent sharply in any direction, partite; genital pore slightly closer to acetab- usually commencing postacetabular (up to ulum than cecal bifurcation; ovary 165 by 140, 175) but may begin dorsal to latter's posterior 95 postacetabular; seminal receptacle 186 by one-fourth or as in one specimen anterior to 167, posterodorsal to and larger than ovary; its micllevel (lying obliquely at its antero- right vitelline field 410 long, left 435; right dextral margin with proximal portion over- field extending 325 postacetabular, left 315; lapping right cecum dorsally), anteriormost five eggs 39 to 41 by 20 to 22. extent 48 to 110 preacetabular; cirrus sac con- DISCUSSION: Considerable individual varia- taining long, relatively thin walled, tubular to tions occurred as noted for the population of saccular, occasionally bipartite seminal vesicle, 63 worms from the kidney of a single cobra. short pars prostatica, prostate cells, and long, Depending upon the combination of character- very thick, muscular, protrusible cirrus; genital istics they keyed to S. serrata Looss, 1899, or pore on midline or to its left at any level be- S. nicolli Bhalerao, 1936, in the key given by tween cecal bifurcation and acetabulum; ovary Bhalerao (1936), to the latter two, S. lachesidis smooth, 115 to 212 by 114 to 177, up to 115 MacCallum, 1921, or S. renalis in the keys by postacetabular or may overlap as much as Byrd, Parker, and Reiber (1940) and by posterior one-seventh of latter; seminal recep- Skrjabin and Antipin (1961), to S. serrata, tacle 77 to 168 by 85 to 169, usually smaller S. nicolli, or S. renalis in the key by Dawes than ovary but may be same size or slightly (1941), and to the latter three species or larger, usually posterodorsal to ovary but may S. sim-plexa (Byrd, Parker, and Reiber, 1940) be entirely dorsal or posterior; metraterm thick Dawes, 1942, in the key by Dawes (1942). walled, muscular, relatively straight, sur- In all keys the single worm from the small rounded by gland cells, commencing dorsal intestine of the second host keyed to S. najae to approximate midlevel of acetabulum, as- described by Nicoll (1912) from the ureters cending left of cirrus sac, usually extending of Naja naja (syn. N. tripudians Merr.) from slightly anterior to latter before curving pos- India. The 63 specimens appear closest to teroventral to open into genital atrium; vitel- S. renalis described by Tubangui (1933) from line follicles usually large but may be relatively the kidney of Python reticulatiis (Schneider) small, 8 to 22 in each field; fields narrow to broad in width, commencing at level of pos- (Boidae) from Luzon, and to S. nicolli de- terior two-thirds of acetabulum, terminating at scribed by Bhalerao (1936) from the intestine level of body of anterior testis but may end at of Ptija-s mucosus (L.) (syn. Zamenis m. L.) level of its anterior margin; right vitelline field (Colubridae) from India. Our specimens differ

Copyright © 2011, The Helminthological Society of Washington JANUARY, 1967] HELMINTHOLOGICAL SOCIETY 109 from S. nicolli in having fewer and much larger to right cecum 390 to 680, to left cecum 140 vitelline follicles, in not being constricted at the to 550; testes tandem to diagonal, smooth to acetabular level, and in possessing a much slightly lobed; anterior testis 157 to 270 by wider body. They differ from S. renalis in 186 to 405, overlapping acetabulum 15 in having its acetabulum farther posteriorly and one and 145 to 680 postacetabular in other in possessing a prepharynx; no doubt a restudy six; posterior testis 182 to 335 by 180 to 405, of the latter species will show a very short pre- 140 to 1,175 postacetabular; cirrus sac 225 pharynx present. We believe our specimens to to 635 (longitudinal extent) by 73 to 135, be S. renalis, including the one from the small entirely preacetabular to considerably over- intestine; the latter appears different to some lapping latter, nearly straight to much re- extent possibly because of its development in curved; ovary 123 to 235 by 123 to 260, the intestine rather than the kidney. As Nicoll usually partly dorsal to acetabulum but may (1912) noted for S. najae his specimens, "al- be entirely dorsal or postacetabular; right vitel- though mature, are possibly not fully grown." line field commencing 115 anterior to posterior When fully grown they may very well resemble margin of acetabulum or up to 245 postacetab- S. renali-s. In other words, the latter may be ular, left commencing 142 anterior or up to 395 a synonym of S. najae, but this can not be postacetabular; lateral vitelline fields separate ascertained at present due to a lack of com- posteriorly or may be confluent; posterior ex- parative materials and knowledge of their life tremity to right vitelline field 11,0 to 325, to cycles. left field 95 to 180; 13 eggs 75 to 94 by 34 to 51. FAMILY ENCYCLOMETRIDAE DISCUSSION: Yeh (1958) reviewed the genus Encyclometra colubrimurorum Encyclometra Baylis and Cannon, 1924, recog- (Rudolphi, 1819) Dollfus, 1929 nizing three species: E. colubrimurorum, E. HOSTS: Enhydris plumbea (Boie), Elaphe japonica Yoshicla and Ozaki, 1929, and E. rufodorsata (Cantor) (Colubridae). asymmetrica Wallace, 1936. Dollfus (1963) HABITAT: Small intestine. recognized only two of these, declaring the LOCALITIES: San-lun Village, Matsu Island second a synonym of the first. The recovery (E. plumbea}; 12 miles southwest of Seoul, of E. colubrimurorum from Matsu Island, lo- Korea (E. rufodorsata). cated off the coast of mainland China near DATES: 1 August (E. plumbea), 17 October Amoy, represents a new geographic distribu- (E. rufodorsata) 1961. tion record; Elaphe rufodorsata is a new host SPECIMENS: USNM Helm. Coll. No. 61704 species. It has been reported previously in (three slides with one specimen each from E. Enhydris plumbea from North Borneo by us plumbea); No. 61705 (one slide with one (1965) and from Formosa (as Encyclometra specimen from E. rufodorsata). microrchis Yainaguti, 1933). MEASUREMENTS AND SOME PERTINENT DATA FAMILY HARMOTREMATIDAE (based on eight adult and two immature specimens from E. plumbea, six adults measured; Harmotrema eugari Tubangui and one adult from E. rufodorsata, measured): Masilungan, 1936 Body 1,955 to 5,460 by 575 to 1,375; fore- HOST: Cerberus rhynchops (Schneider) body 450 to 1,875, hindbody 1,217 to 3,050, (Colubridae, syn. Homalopsidae). preoral body 39 to 56, posttesticular space 735 HABITAT: Small intestine. to 1,540; oral sucker 255 to 450 by 300 to LOCALITY: Zamboanga, Mindanao Island, 502; acetabulum 315 to 535 by 335 to 585, Philippines. center at level of anterior three- to four-tenths DATE: 21 December 1961. of body length; sucker length ratio 1:1.08 to SPECIMENS: USNM Helm. Coll. No. 61706 1.24; pharynx 157 to 252 by 121 to 295; short (three slides with one specimen each). prepharynx and esophagus present; cecal bi- MEASUREMENTS AND SOME PERTINENT DATA furcation 60 to 425 preacetabular; ceca sub- (based on two complete specimens and one equal in length in nine specimens, equal in with part of preacetabular body missing, from two, right usually shorter, posterior extremity one host): Body 2,266 to 3,171 by 565 to 630;

Copyright © 2011, The Helminthological Society of Washington 110 PROCEEDINGS OF THE [VOL. 34, No. 1 forebody 420 to 930, hindbody 1,735 to 2,125, vesicle may be saccular to tubular and coiled, posttesticular space 595 to 790, postcecal space occasionally bipartite in appearance, rather 245 to 285, postvitellarian space 195 to 215; than saccular as illustrated, and the metraterm oral sucker 90 to 97 by 73 to 85; acetabulum commences at the anterior margin of the an- 111 to 116 by 88 to 118, center at level of terior testis and is winding rather than anterior anterior one-fourth to one-third of body length; to this level and being straight as illustrated. sucker length ratio 1:1.20 to 1.23; pharynx 80 by 73 to 77; esophagus 22 to 80 long; testes FAMILY OPHIODIPLOSTOMATIDAE 109 to 222 apart, longitudinally or transversely Proalarioides kobayashii Park, 1940 elongate, smooth to slightly irregular in out- HOST: Dinodon rufozonatum (Cantor) (Co- line; anterior testis 145 to 240 by 184 to 242, lubridae). 500 to 740 postacetabular; posterior testis 182 HABITAT: Small intestine. to 259 by 155 to 217, 790 to 1,195 postace- LOCALITY: 12 miles southwest of Seoul, tabular; cirrus sac 179 to 345 (longitudinal Korea. extent) by 95 to 177, equatorial with anterior- DATE: 17 October 1961. most margin 260 to 500 postacetabular, thick SPECIMENS: USNM Helm. Coll. No. 61707 walled, muscular, containing seminal vesicle, (four slides with one specimen each). short pars prostatica, prostate cells and long, MEASUREMENTS AND SOME PERTINENT DATA spined cirrus; seminal vesicle saccular when (based on 11 mature and three young adults, distended with sperm to somewhat tubular and and 22 immature worms from one host, seven winding, occasionally appearing bipartite; gen- mature measured): Body 2,001 to 4,027 in ital pore 375 to 570 postacetabular; ovary 92 total length; spherical forebody 1,089 to 2,078 to 131 by 114 to 162; metraterm very thick by 1,035 to 2,048; cylindrical hindbody 897 to walled, muscular, somewhat winding, com- 2,038 by 495 to 1,380; anterior extremity of mencing at anterior margin of anterior testis; body to acetabulum 391 to 620; posttesticular five operculate eggs 104 to 111 by 63 to 77, space 468 to 1,598; postcecal space 376 to few. 1,414; oral sucker 90 to 148 by 140 to 210; DISCUSSION: Tubangui and Masilungan acetabulum 115 to 175 by 148 to 197; tri- (1936) described this species from Luzon bocytic organ 475 to 767 by 422 to 843; Island, Philippines, listing the host as a cobra, pharynx 106 to 136 by 70 to 116; esophagus Naja sp. (Elapidae); later, Tubangui (1947) 36 to 175 long; testes 34 to 178 apart; anterior specifically identified the host as Naja naja testis 152 to 250 by 213 to 302, 552 to 2,294 philippinensis Taylor. Our report represents postacetabular; posterior testis 150 to 230 by new host and geographic distribution records. 206 to 360, 736 to 2,669 postacetabular; her- Yamaguti (1958) and Skrjabin (1962) placed maphroditic sac present; ovary 97 to 194 by the genus Harmotrema Nicoll, 1914, in the 111 to 195, 499 to 2,117 postacetabular; 17 family Liolopidae Dollfus, 1934, and subfam- operculate eggs 98 to 114 by 48 to 65. ily Harmotrematinae Yamaguti, 1933; Mehra DISCUSSION: Park (1940) described this spe- (1962) erected the family Harmotrematidae cies from the vicinity of Seoul, Korea, from for this genus and Helicotrema Odhner, 1912. Elaphe dione (Pallas) (Colubridae), Natrix Our specimens show several variations from tigrina (Boie) (Colubridae), and Ancistrodon the original description of H. eugari. In our blomhoffi brevicaiidiis Stejneger (Crotalidae). forms the left cecum is decidedly shorter than Our report is a new host record. Yamaguti the right rather than nearly equal in length (1958) placed the genus Proalarioides Yama- as illustrated, the acetabulum is at the level of guti, 1933, in the family Proterodiplostomidae the anterior one-fourth to one-third of body Dubois, 1936, and subfamily Ophiodiplostom- length rather than near the anterior one-sixth, inae Dubois, 1936; Sudarikov (1960) erected the intertesticular space is shorter than the the family Ophiodiplostomatidae and subfam- length of either testis rather than longer, the ily Proalarioidinae for this genus. Six of our testes may be wider than long rather than al- seven measured specimens are shorter in total ways longer than wide, the cirrus sac is equa- length than the shortest noted by Park. In torial rather than preequatorial, the seminal spite of this the postcecal space in our forms

Copyright © 2011, The Helminthological Society of Washington JANUARY, 1967] HELMINTHOLOGICAL SOCIETY 111 is much longer, averaging 826 microns, rather HOST: Hemidactylus frenatus (Gekkonidae). than being between 240 to 250. Additionally, HABITAT: Stomach. the acetabulum is located much farther pos- LOCALITY: Clark Air Force Base, Luzon teriorly, averaging 513 microns from the an- Island, Philippines. terior extremity of the body, rather than being DATE: 15 December 1961. between 140 to 150. SPECIMENS: USNM Helm. Coll. No. 61710 (three slides with one specimen each). FAMILY DICROCOELIIDAE Measurements and some pertinent data Paradistomum gregarium Tubangui, 1929 (based on three specimens from one host): SYNONYMS: Paradistomum magnum Tuban- Body 1,565 to 1,882 by 815 to 950; spines gui, 1928, nee Travassos, 1919; Paradistom- sparser posteriorly, lacking at extreme pos- oides gregarium (Tubangui, 1929) Travassos, terior end of body; forebody 415 to 515, 1944. hinclbody 958 to 1,195, previtellarian space HOSTS: Hemidactylus frenatus Dum. and 202 to 295, posttesticular space 480 to 705, Bibr., Platyuriis platijunis (Schneider) (Gek- postcecal space 225 to 410; longitudinal rows konidae). of subcuticular parenchyma! gland cells in fore- HABITATS: Gall bladder, liver. body; oral sucker 157 to 169 by 157 to 172; LOCALITIES: Tacloban, Leyte Island (H. acetabulum 157 to 170 by 152 to 160, center frenatus, P. platijunis); Clark Air Force Base, at level of anterior three-tenths to one-third of Luzon Island (P. platijunis):, Philippines. body length; sucker length ratio 1:0.99 to 1.02; DATES: 14 (Leyte), 21 (Luzon) December prepharynx present, short; pharynx 76 to 94 1961. by 61 to 75; esophagus sinuous, as long as or SPECIMENS: USNM Helm. Coll. No. 61708 longer than pharynx; ceca conspicuous because (four slides with one specimen each from H. of very thick cell lining, ventral to testes; left frenatus); No. 61709 (five slides with one (anterior) testis 390 to 440 by 295 to 385, specimen each from P. platijunis). right (posterior) testis 390 to 450 by 378 to DISCUSSION: Our collection consisted of 398; cirrus sac 254 to 290 by 140* to 145, four, five, six, and seven mature adult worms relatively thin walled; pars prostatica with from four H. frenatus, and one (mature adult, conspicuous cell lining; genital atrium shallow; in two), three (mature adult), four (mature genital pore in one worm slightly posterior to adult), 14 (two mature and nine young adult, level of posterior margin of acetabulum, over- three immature), and 17 (seven mature and lying anterior part of left testis and lying 10 young adult) from six P. platijunis. The anterior to posterior margin of acetabulum in measurements and data, including extensive other two; ovary 170 to 220 by 205 to 245; morphological variations, for the present speci- uterus may extend anteriorly as far as anterior mens are essentially similar to those reported margins of ovary and acetabulum; metraterm by us (1964, 1965) from H. frenatus from long, sinuous, thick walled, muscular, sur- Palawan Island (Philippines) and North Borneo rounded by prominent gland cells; 15 oper- (Malaysia). This species was originally de- culate eggs 19 to 23 by 11 to 13. scribed from H. frenatus from Luzon, and DISCUSSION: Tubangui (1928) described redescribed from H. gleodovi Murray from this form as a new genus and species from Burma; we (1965) found it in Gehyra mutilata the same host and island as our specimens. (Wiegmann) (Gekkonidae) from North Bor- The latter show several variations from the neo. Its presence in Platijunis platijunis and original description. In our forms the acetab- location on Leyte represent new host and geo- ulum is about the same size as the oral sucker graphic distribution records. rather than being smaller, the esophagus is relatively long rather than being "very short, FAMILY LECITHODENDRIIDAE almost nil," the genital pore may be posterior Postorchigenes ovatus Tubangui, 1928 to the level of the posterior margin of the SYNONYMS: Palitrema macrorchis Gogate, acetabulum rather than anterior to this margin, 1939; Postorchigenes macrorchis (Gogate, and the testes are larger in size while the 1939). ovary is the same size or smaller. Tubangui

Copyright © 2011, The Helminthological Society of Washington 112 PROCEEDINGS OF THE [Voi,. 34, No. 1 made no mention of the presence of a pre- adultes chez des batraciens, reptiles, oiseaux. pharynx or metraterm. There are two other Ann. Parasit. 38: 29-61. recognized species in the genus: P. macrorchis FISCHTHAL, J. H., AND R. E. KuN'rz. 1964. A monogenetic and seven digenetic trematodes (Gogate, 1939) (syn. Palitrema m. G.) from of amphibians and reptiles from Palawan lizards, Hemidactylus brooki Gray (Gekkon- Island, Philippines. Proc. Helminthol. Soc. idae) and Calotes versicolor (Daudin) (Agam- Wash. 31: 230-240. idae) from Burma, and H. flaviviridis Rueppell , AND . 1965. Digenetic trematodes from India; P. duboisi Rohde, 1963, from a bat, of amphibians and reptiles from North Bor- Cheiromelas torquatus Horsfield (Molossidae), neo (Malaysia). Ibid. 32: 124-136. from Malaya. Our material fits the descrip- COGATE, B. S. 1939. On a new trematode Pali- tions of P. macrorchis given by Gogate (1939) trema macrorchis, gen. et sp. nov. from Ran- goon lizards. Rec. Indian Mus. 41: 57-60. and Baugh (1957). The specimens illustrated HUXLEY, J. S. 1932. Problems of relative growth. by the former from H. brooki and the latter 276 p. Dial Pr., N. Y. from H. flaviviridis are more like ours than is Li, L. Y. 1937a. Some trematode parasites of that shown by Tubangui. The longitudinal frogs with a description of Diplodiscus sinicus rows of gland cells noted by Gogate and by sp. nov. Lingnan Sci. J. 16: 61—70. Baugh were seen only in part in one of our . 1937b. On the excretory system of specimens. It has been our experience and Glypthelmins staffordi Tubangui, 1928. Ibid. that of other investigators that the detection 16: 303-305. of parenchymal glands when present depends MEHRA, H. R. 1962. Revision of Brachylaem- idae Joyeux et Foley, 1930 with new sub- upon the method employed for preparation of families Thapariellinae and Urotrematinae and the whole mounts and are not always demon- new family Harmotrematidae with its sub- strated. We declare Postorchigenes macrorchis families, Harmotrematinae Yamaguti, 1933 (Gogate, 1939) a synonym of P. ovatus. and Helicotrematinae n. subf. Proc. Natl. Acad. Sci. India, Sec. B 32: 319-334. LITERATURE CITED NASIH, P. 1966. Two new species of digenetic trematodes from Venezuelan amphibians. BAUGH, S. C. 1957. Contributions to our knowl- Proc. Helminthol. Soc. Wash. 33: 166-170. edge of digenetic trematodes II. Proc. Natl. NICOLL, W. 1912. On two new trematode para- Acad. Sci. India, Sec. B 26: 295-313. 1956. sites from the Indian cobra. Proc. Zool. Soc. BHALERAO, G. D. 1936. Studies on the hel- London 1912, No. 4: 851-856. minths of India. Trematoda II. I. Helm. 14: PARK, J. T. 1940. Trematode parasites of Rep- 181-206. tilia from Tyosen. I. Three new digenetic BRAVO H., M. 1941. Revision de los generos trematodes, Encyclometra koreana sp. nov., Diplodiscus Diesing, 1836 y Megalodiscus Neomicroderma elongata gen. nov. sp. nov. Chandler, 1923 (Trematoda: Paramphistomo- (Plagiorchidae) and Proalarioides kobayashii idea). I. An. Inst. Biol. Mex. 12: 127-146. sp. nov. (Strigeidae). Keizo J. Med. 10: 113- BYRD, E. E., M. V. PARKER, AND R. J. REIBER. 123. 1940. Taxonomic studies on the genus Sty- ROHDE, K. 1963. Uber einige malayische Trem- phlodora Looss, 1899 (Trematoda: Styphlo- atoden. Ztschr. Parasitenk. 22: 268-277. dorinae), with descriptions of four new spe- . 1966. On the trematode genera Lutz- cies. Tr. Am. Micr. Soc. 59: 294-326. trema Travassos, 1941 and Anchitrema Looss, DAWES, B. 1941. On Styphlodora elegans n. sp. 1899 from Malayan bats, with a discussion of and Styphlodora compactinn n. sp., trematode allometric growth in helminths. Proc. Hel- parasites of Python reticulatus in Malaya, with minthol. Soc. Wash. 33: 184-199. a key to the species of the genus Styphlodora SKRJABIN, K. I. 1962. Family Liolopidae Doll- Looss, 1899. Parasitology 33: 445-458. fus, 1934. In Skrjabin, K. I. [Trematodes of . 1942. A new name for Styphlodora animals and man]. Moskva. 20: 7-26. (Rus- compactiim Dawes, 1941, a criticism of the sian text.) proposed genus Paurophylltim Byrcl, Parker , AND D. N. ANTIPIN. 1961. Family Pla- & Reiber, 1940, and a revised key for the giorchidae Liihe, 1901. Part 4. Ibid. 19: 195- separation of species of the genus Styphlodora 266. Looss, 1899. Ibid. 34: 266-277. SUDARIKOV, V. E. 1960. Suborder Strigeata La- DOLLFUS, R. P. 1963. Mission Yves-J. Golvan et Rue, 1926. Part 3. Ibid. 18: 453-694. Jean-A. Rioux en Iran. Trematodes Digenea TUBANGUI, M. A. 1928. Trematode parasites of

Philippine vertebrates. Philippine J. Sci. 36: I. The digenetic trematodes of vertebrates. 351-371. Parts I and II. 1,575 p. Interscience Publ., . 1933. Trematocle parasites of Philip- N. Y. pine vertebrates, VI. Descriptions of new , AND Y. MITUNAGA. 1943. Intestinal species and classification. Ibid. 52: 167-197. helminths from Bufo melanostictns of For- . 1947. A summary of the parasitic worms mosa. Tr. Nat. Hist. Soc. Taiwan 33: 142- reported from the Philippines. Ibid. 76: 225- 154. 322. YEH, L. S. 1958. A review of the trematoclc , AND V. A. MASILUNGAN. 1936. Trem- genus Encijclometra Baylis and Cannon, 1924. atode parasites of Philippine vertebrates, VIII. J. Helm. 32: 99-114. Flukes from a cobra and a crocodile. Ibid. YUEN, P. H. 1962. Three trematodes from Ma- 60: 255-265. layan amphibians including two new species. YAMAGUTI, S. 1958. Systema helminthum. Vol. J. Parasit. 48: 532-535.

Presentations 1966 ANNIVERSARY AWARDS OF THE HELMINTHOLOGICAL SOCIETY OF WASHINGTON 421st Meeting 19 October 1966

George Roger LaRue, whom we honor at Among other prior positions were Science this meeting with an Anniversary Award of Assistant and Assistant Zoologist, U. S. Bureau the Helminthological Society of Washington, of Fisheries, honorary Curator and Research is recognized as one of the most influential Associate, University of Michigan Museum, and respected American zoologists and para- and Fellow by Courtesy, School of Hygiene sitologists of his generation. and Public Health, Johns Hopkins University Born in Paullina, Iowa (1882), he was grad- (1924-1925). uated from Doane College (B.S., 1907) and He received an American Medical Associa- received his graduate training at the University tion honorary Research Grant and an honorary of Nebraska (A.M., 1909) and the University Sc.D. from Doane College (1947). of Illinois (Ph.D., 1911) under Professor H. B. During his long tenure at Michigan, he Ward. trained and served as advisor to numerous After service as Assistant Zoologist, Doane graduate students who subsequently achieved College (1905-1907), Technician, Medical leadership as educators and researchers in College, University of Nebraska (1907-1909), zoology and parasitology. While affiliated with and Research Assistant and Assistant Zoologist, the U. S. Department of Agriculture at Belts- University of Illinois (1909-1911), he em- ville, he was a valued consultant and advisor barked on a distinguished career at the Uni- to his associates, ably reviewed many of their versity of Michigan. It began in 1911 and research manuscripts and presented an emi- officially spanned 41 years. He was Professor nently mature graduate course in trematoclol- of Zoology for 25 years, Chairman of the Zool- ogy in the Departments' Graduate School. ogy Department for 15 years and Director of Much of his research, which resulted in the University's Biological Station at Douglas many valuable contributions to knowledge of Lake for 23 years. He was appointed Visiting the morphology, taxonomy, and life histories Investigator, Zoological Division, U. S. Depart- of tapeworms and digenetic trematodes, was ment of Agriculture, Beltsville, Maryland, in conducted at the Michigan Biological Station 1951, Emeritus Professor of Zoology, University where his prominent position caused students of Michigan, in 1952, and Visiting Professor of and faculty to name him "Governor of North- Parasitology, Rice Institute, for 1952-1953. ern Michigan."

Dr. George Roger LaRue (left) receiving a 1966 Anniversary Award of the Helminthological Society of Washington (presented by A. Mclntosh).

Previously honored with the presidency of and in token of our esteem for him and appre- the American Microscopical Society and the ciation of his 53 years of loyal membership, vice-presidency and presidency of the Amer- The Helminthological Society of Washington ican Society of Parasitologists, he ably served is privileged to present to George Roger LaRue as Editor of the Journal of Parasitology in 1954 this Anniversary Award. (Committee: Lucker, and 1955, when more than three-score and ten, Lund, Sadun. Presentation: Allen Mclntosh.) and is still a Consultant to the Editor. Early in 1913, together with such notables William Walter Cort, now to receive an as Theobald Smith and S. T. Darling, he was Anniversary Award of the Helminthological elected an American corresponding member of Society of Washington, is noted and esteemed the Helminthological Society, then just over internationally as a medical helminthologist two years old. In 1924 he participated in the and parasitologist. actions of the Society which resulted in the He was born in Leon, Iowa (1887), is a founding of the American Society of Parasitol- graduate of Colorado College (1909), and ogists. He has regularly participated in affairs received an A.M. (1911) and a Ph.D. degree of the "Helm.-Soc." since 1951 and was elected (1914) in Zoology from the University of Illi- an Honorary member in 1959. nois, where he studied under Professor Henry To honor him for his far-reaching influence B. Ward. as an educator and molder of the professional He was Instructor in Zoology at Colorado careers of now leading zoologists and parasitol- College (1912-1913), Professor of Zoology at ogists in this country and abroad and his ac- Macalester College (1914-1916), Assistant complishments in helminthological research Professor of Zoology at the University of Cali-

Dr. L. J. Olivier (left) aecepting on behalf of Dr. William Walter Cort a 1966 Anniversary Award of the Helminthological Soeiety of Washington (presented by J. T. Lucker). fornia (1916-1919), and in the School of Public Health Service; Director, Program of Hygiene and Public Health, Johns Hopkins Study of Ascariasis in Children, National Re- University, became Associate Professor of Hel- search Council and American Child Health minthology (1919-1925), Professor of Helmin- Association; Member, Commission on Hook-- thology (1925-1943), Professor of Parasitology worm Diseases, International Health Division, (1943-1953) and Emeritus Professor (1953-). Rockefeller Foundation. He was appointed Research Professor (1953- During his tenure at Johns Hopkins, dozens 1959) and Emeritus Research Professor of graduate students earned a doctorate under (1959-) in the School of Public Health, Uni- his tutelage and guidance. Many subsequently versity of North Carolina. achieved leadership in research in medical or An honorary Sc.D. was awarded to him by veterinary helminthology. the University of North Carolina (1946) and Also notable is his enviable record of re- by Colorado College (1949). search accomplishments in helminthology. He Among other positions in which he also has investigated the morphology, embryology served were: Consultant Helminthologist, Cali- and life cycles of trematodes, hookworm dis- fornia State Board of Health; Visiting Professor, ease, ascariasis, schistosomiasis, and many other Peking Union Medical College, China; Lec- problems. He is the author, or a coauthor of turer, School of Public Health, Harvard Uni- about 200 parasitological books, monographs, versity; Tropical Medicine Consultant, U. S. papers and notes published from 1912 to 1960. Secretary of War; Consultant, Laboratory Di- A member of several professional societies vision, Communicable Disease Center, U. S. and active in all, he was honored with the

Copyright © 2011, The Helminthological Society of Washington 116 PROCEEDINGS OF THE [VOL. 34, No. 1 presidency of four, among them the Amer- his students to join the Society and was a ican Society of Parasitologists, which he also member of the "Helm.-Soc." committee which served as Editor and in almost all other official implemented the founding of the American capacities. Society of Parasitologists and even provided Elected a member of "Helm.-Soc." in 1920, the constitution adopted by the founders in he has since been an active participant in its 1924. affairs. He was elected President for 1924- In recognition of his high rank as an 1925 and a life member in 1954. Apparently, authority, educator, researcher, and instigator he sometimes made the roundtrip from Balti- of research, in medical helminthology and in more to Washington meetings by railroad and grateful appreciation of his contribution to streetcar! He and his associates at Johns Hop- this Society's stature, welfare and growth, kins were hosts for all November meetings The Helminthological Society of Washington from 1920 through 1934 and for many subse- proudly presents to William Walter Cort this quent Baltimore meetings; often supper was Anniversary Award. (Committee: Lucker, provided for all comers. He induced many of Lund, Sadun. Presentation: J. T. Lucker).

In Memoriam MOHAMMAD ABDUL BASIR Aligarh Muslim University 1916-1966 Member since 1948

/-?'"' ^_ (Alabama through'Maryland;;rremainder of list will appear-in Jtdy issue) , „/•

Alabama ' Ericksbo, D. G. --Ferris, "Virginia R. Cairns,E. IT '.'- Habermann, R.^T. Seitner, P. C. .HerroaiLC. M. .. /-', FjandsenTj. C. : Hernandez. A.-;:-' - Shumard, R. F. Isenstein, R. $., HopeTW.l). : > ; Thorson, R. E. Holbrook/A. A. Rogersrw."Av Kenworthy,-F. T. Iowa v " '' -Huff, C. G. - ~; ^ Wellborn, T. i., Jr. lancicome, D. R. EUis,C.X ^ ^'Humphrey, Judith M. Âlaska Martin, L. K, Epstein, A. H.-v :,:,: ^Jachowski, L. A., Jr. v -" Rausch, R.—"•.. '- - McMullen, D, B. Goss^, R,G^ 7'-''' I : Jacobs, L. - • .,." . /-• t '. Arizona Olivier-rL. J. x Greve, J. H. . Anderson. G-Ar Hsii.H. F.- Kates, K.-C. ' V " Redington, B. C. Norton, D. C- Knight, R. A. . •--•_ Reynolds, Hr,W. Reid, W. A., Jr. - ^ UhnerrM. J. , ^ Knisberg, L. R. /Arkansas Sadun,E. H. Vande Yusse, F, J. / LaRue,G.R. -~r.' Slack, D. A.- - ..SlieVRlta-B..-""•>. Zimmermann, W. J. .^.Lawless, D. Kr ~ California ". Tiner.J.D: ." Lieek, R. G. -- ^ Allen. M. W.~ -O Florida ^ - Ackert, J.Ê. ! • • 'LeJseXj-E. —•• '-•':•' — ' ^' Ayoub, S. M. -; / Christie, J;;JL -^ ;• Ameel^D. J. Ir C^ .Uchtenfels, J; R. r..; / Baker, N. F, / Aytan, Songul "" ~^~ /Lotze; J.41 - v • Brown,R.,L. ^.-^ . Esser, R. P.' s./~'^ Coil.W. H, ,:/ , Lucker, J. T. ^ _ ^; : Caveness, F.T3." Hannpn, C. L.' -^ Dickerson, O, J. ' Lund, E. E. -' _ s' •'•' " J. D., Hunter, G. W. HI •Hansen, M. F. : /Lunde, M.;N. - '^T^" .^ Kincaid, R. R. , 'Kentucky . , ^-.''<_'.^ 'Luttennoser, G. W. Hart, Mary Ann- O'Bannon.jJ. H. Chapman, ;R. A. _ _- flart,'W,H. -;. s Overman, Amegda J. ;Eversmeyer,,H. EL--- Marchbank, Dorothy F. Heyneman, D. - 'Perry, V. G. ;; McIritosh,-A. -'/r Holdemaii, Q. 1* , Bhoades, H. L. •.-•' - Louisiana ' ,.'"->~ McKinStry.D. M. ' . Konicek, DÊ. T* 'Shortj^R. B.~ -=-c Acholonu, A. D. ---^ McLoughlin, D. K. Krassner, S. M. SmartT G. C., Jr. - ^Birchfleld. W. ; Morris, J. A. . Tarjan.A.C. AHolhs, J. P. JTrr :-,. Murpby,/D. G. • '.-.-."• -l^mifejight, H. W. Taylor. A. L. -.-.'"- Ibrahim, I. K. A.- Nickle, W. Rr^f Xie-KIan-Joe ~" Tomerlin, AGiordia. H. - /" - •Sogandares-Berriffl,-F. Pipkin, A. C. • • T >McBeth, C:;W. 2: Cross, H. F. - Warren, L. G. r^ Policy, Judith A. McGuire, W. G. Denton, J. F. /: Maine • - Mullee, Mary/T. Goodchild, C. G. Powers, K. G. _ ; ^Heard,'R.!W., Jr. ^Maryland ^Richaids, C, S.~ Noffsinger, Ella Mae Kagan, I. G. Amerault, T. E. : Roby, T. O. - •••' Parinan,\G. A. /"Mi)iton.;N. A. • J • :"Ana»tos,.G.. -'Rose,;J.iE.- -/ ".i- Poinar, G. O.,-Jr. ? Price, C.E. . /Andersbn,'R. I. K , L, E. -iVuehie,;T.XL. < " . Andrews, J.S. " Rossan, R.^N. ^Stewart, T.B. . -; ; Ânthony,:D. W. : Sawyer, T. K. Rothman, Al;H. Hawaii / i Armstrong, D. E. Sayre,R/M. Sher.S. A. I.E. 'iSteele, A. E. ,^- V ;:BecHund,W. W.- ^. L. - ' _: ; Van Guiiidy, •». rD. "/Cheng, T.C. - -Bergner, j. F., Jr.- Schneiderr:C.?M. xViglierchio, D. R. Goto, S. ;; '£ , von Brand, T. - Seott, J- A." - Voge,-Marietta 'Buhrer, Edna M. /Segal, Dorothy B. ;Wagner,E./D. Idaho' --^ ." /Cannon, L.'T. - - Sheffield, H/G. ''Weinmann,-, C. 'T- DalKmore, C. E. /-/Castillo, Jessica>l. Shiroishi, /Tsugiye'-- -- White, L; V. Illinois ,Ghitwood,-May B. Sfaorb;>D.Ar ' ~ ; Canal Zone ' j- Bauman,P.-M. Colglazier, "M. L. Walton, B.C. iDunagan, T. T. *&'. ; -DeWitt, W;^B. ' i /.Soloniori, G.:B. :"._.-/ 'Colorado Edwards, 15. I. -Diamond; L. S.- Spindler, • L. A. - ;' Olsen, O. W. JK^ i} Garoian, G. ^ Doran.D. J-~ Sprague, V. , -- . - 'Schmidt, G.^D. /Hechler, Helen C. Doss, Mildred Aî- Stirewalt, Margaret^ ~:r-~. Stabler, R. M. - - Kruidenier, F. J.. - Douyres, F. W. , Sylk^.^R; '- - :/ Ubelaker, j. E. ,_ 3aevirie, N.:D. / /Dufbin; C. G. /Tarshis, I. B. :" .7 ." Connecticut. • '•' McCrae, R; C.:- .Êndo,;B.Y. ::~ -Terzian,' Lv A. 'Miller, P.?M.-- Ostdiek, J.^L. Traub'.R. -" : ^~ ;' Peiiner, L; R,-"•--'— , Pillai,J. K. ./- Enzie, F.-D..7"/';- -TRromba, i?."G.-~~ ----- "~ Delaware Schneider, M. D, ;. - Farr, -MarionsM.' " /Turner, J. H. ~^'- - Fielding, M.J. Siddiqui, ;I. A. Feldmesser, J. ~."-. /Underwood, P. C. 'District of Columbia ' Singer, I. 4 : ^Ferguson, M.rS.//; Baisden, L.:A. ---. Taj^lor, D. P. ,Betz, D. O., Jr. /; Thomas, L, Jr Foster,'A.'O.' * ; Wehr, E. E. —Briggs,'N.yT.:.?, ---j; sWalton, Ar

'"'''•''•'". -'-~-; • -"•' - '">, '{ v -~ ' C"-'i y' i : ''' v' V" "X.f, • '••/''-'-'.' X ,-.• • ""••' ' "'" - vTTL •" -.: v • •'•' \-X- '\r~;-^"\: '•" I -i • • ,\X - ^-VOLUME 34 - ''.-F/;X X^-.XX JANUARY 1967 f^^-.{---\' NUMBER It f •' 7-X^ --•'• /X^':\X, XXX7.7.xXX..; '•;;..:;",X-v\ X-vr"",'-'.-- X --- : • •, 'X'[" -X '">7~- ".'/;"- -' •, :'• •• .-X''J'-1

'|f3Bfeas, yracmiA R.; Ahro JOHN M. "EjEatt^ ' f r. , >/oc 104 FISHER, J. M., J(ND D. J, RASKI. \g of Xiphinema index and X diversicaudatum 68 Encholainioidea (Nematoda: Dorylaimida), New Superfanuly Representing .Dorylaimid Specimens\wijth ^Cephalic Setae .,_„:;„ .94 ? X ;X HOOEVVIND, WHAIA L., -AND J. JHETOS. Iôtes on the CJenus Isolaifftium Gobb, 1920 Pj ) X" . fNematoda), with Descriptions of New Species from Sbu& Africa __-.._.__. ____ .,.— 20 ' ^- HOPE, W/DUANE. A Review of ^ the ; 'Gehuis PseudoceUa Fftipjev, 1927 (Nematoda; ; x ; -" - 7 CeptosomatidaeJ.mlija /Description of Pfi^doceZIo frwtdo&^tw n. ^/ ^-^^-.-^, . B , ; x £ HXJNTEB, WAT«JA SANBOKN. Notes on the. %,ife, History of jPfeurogomtw malaclemys \. . \ X" ^ x_" •Hilnjet, 1961 (Trematoda: PronOcephaUdae) -from Beauiort, , 'North Carolina, with ^ r V'^X'^,..-1'. X; ( !a Description o^ the iGercaria X^-. — -X.^— - — —:._ - —^^.^'I^-ZL : — ...___.„.._„._.••._ ;;33:: IBRAHDCM, I.^K- A. Morphological ^Differences 'Between tiie 'Cuticle of Swarming and - Nonswarriiing Tylenchorhynchtis martini .: ____ :_i.'; ------_-l^__._.^.... X — _lX._....— ____.: _ 18 '-,-' "~ ' -' - ' • ••- ' " , _ . . -'••'''_ - :- __ • , STUART E,,-AND JOSEPH E. AUCATA; failure -^fCertamGlami and Oysters to Serve as Intermediate Hosts for Angiostrongylus cantonensis — ^.— ;.-__:„>,....„______. ^ ; j ICuN'TZ, RoBERT-E.; ^^ ;BETTY Jx-WE MYBRS, AAND THOMAS >E.V\*ICE. Intestinal Protozoans ^ X arid[Parasite s of ;the,Gdada Baboon (fheropithecusgelddaRup^l,IS35}\-^^ ____r 65 JOHN W. \Monorddtaenia ,nom. fjiov. for the ?Badger Taeniid Gestodes One Row of J •" • '<-• - ; •'- ;> / '- • • •••' : , ... • -. - . f" ' • "XivlEYER, MARVIN C., NAND, WILLIAM G. VALLEAU. The Effect of Gonadectomy , and ( l Hormone Therapy in Male Hamsters rUporitihe Egg Output by the Pseudophyflidean - \:-- DiphyUobothrium isebygo ^^l^lii-i*^^^*^^ j- <•/ '"" ' ^'"^v '-' f'' 1-' ~ '--••• \" 'X •' "/"'''""""'-•" ; ' * % ';'"•. ,' • "• -'-> '. - • . , .' " , ">' j- . i N-" -" —- - ^ X.y.^vX NIGKLE, W; R. On the Glassif ication of /this Insect Parasitic Nernatodes of the Sphaer- ;'-^~ X ' '7^'^'. ulariidae l/ubbock, 1861 ^ (Tylenchoidea: JSFematoda) . ______-._-L,^..^-,.: _____ *£*. ______72 • ..-- I,'"'"'. •-•••••'.-';'.,'. '''•:•;•..' -•:."';.-•.'. ,'~ • ',- •""- -•''. "- ."-./:""."-'"' ''''" -.\ ' ' •••:--''_ ' r; Y '- >.- r^ -X j.-"1 ":'..'î -"''.'•'•/•. ;'NyBERG,,,PKrER i^^Q&i^^^^î^^^j^^JhsaJ^^^â^, Incidence 'of -Bovine L" . ' 'XX' ';.:, r^,^ ' Goccidia in':êstern -.Oregpn. ._l...._-_:,—__.._.. - .--^—-,^-..-.;J. — ..^..^.i —l..l_...._.i. ; \: . TSZ '._: ' :r' '•' ~,..^' ;••' '•- -.' ' • ' V ' ,--'' , ;',--, '•''•• ' ' '• "".,""" ••' ' ;: • •_ ' ': - ' \: ' '" ' ' '-."'--'':''. r- X i ~-;VfH Presentations— 1966 Anniversary Awards of The Helminth6logical|Society of Washington ^113 "' ''' ' " '" ' " ' ' * ' ' / X\•'_.^', ROGERS, WILMER A. Myzotrema cyclepti ^gen. n., sp. n. (Trematoda: Monogenea) -: -(^ v -c X"' from Gills of: .CycZepitw^JongofMS (LeJSueur) from Alabama—X--^- --— T-——-•- /3 X ,: - SPINDLEH., L. A. Experimental Transmission of ffiptomonas meleagridis and Hetetal&s ' ; x> v- gattinarum by the SowAug, forcbttio scdber, And Its Implications for Further X ~; X -•.,r;X: ;:;; ; ;: Research- ^&Zi::£$^^ 26 ;-.." ' „,/ TAYLOR,'.~D.' P., AND J/K. PHXAI. Parqphelenchiis acgntioides n. sp. (Nematoda: . ! - - ;. ':-;: /Paraphelenchidae),;a Mycophagous Nematc4efr<)m Illinois, with Observations;pn its : T : ^ , Feeding Habits and a Key(to the Species :oi ParaphelenchiK ^---.~--.^--.^~-.-..^.L^. ~5l X A X- '.L / 7 WELLBORN^THQMAS L., JR. ' Four New Species o^ iGyrodaotylite (Trematoda: M6no-_ : ^cV>J^'-';5:V'.^'v- "geneaO'feoni Southeastern U.(S, .^.^^^^L.^..^..^-^^.^.-...^-.!^,...... ^--—•:.„'__. 55 r'-VT^"^^' " 'v X?;.-r-',-'X ^ _'-> ^-V -•- .-; '-.".'.;', "••'^ v •' _ "''^.iV ' ':.'-'-"X\- - '-'^-''' ' ' ~- XX'--' ' "* t'-iX :.-rX^^( -^;IE 33 i .^7:^.'..' 7-"<-,.. V ...-"•'\/-'X ^y^'-^^^^S^î^ Number. 1, 24March 1966 /-^ '- ~{., Nijraber 2^29Julyi966 7 . >^ [^>.

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