(Nematoda, Cosmocercidae) in Duttaphrynus himalayanus (Amphibia, Anura) from Dehradun (Uttarakhand), India

DOI: 10.2478/s11686-014-0212-8 W. Stefański Institute of Parasitology, PAS Acta Parasitologica, 2014, 59(1), 80 84; ISSN 1230-2821 Cosmocercoides himalayanus sp. nov. (Nematoda, Cosmocercidae) in Duttaphrynus himalayanus (Amphibia, Anura) from Dehradun (Uttarakhand), India Anjum N. Rizvi 1 and Charles R. Bursey 2 * 1 Zoological Survey of India (ZSI), Northern Regional Centre, Dehradun-248 195, Uttarakhand, India; 2 Department of Biology, Pennsylvania State University, Shenango Campus, Sharon, Pennsylvania 16146, USA Abstract Cosmocercoides himalayanus sp. nov. (Nematoda, Cosmocercidae) from the large intestine of Duttaphrynus himalayanus (Amphibia, Anura) from Dehradun, India is described and illustrated. Cosmocercoides himalayanus sp. nov. represents the 21st species assigned to the genus and the 9th species from the Oriental biogeographical region. Cosmocercoides himalayanus sp. nov. differs from the previously described Oriental species in number and position of rosette papillae; it is the only species possessing 24 or more rosette papillae to have 4 postcloacal papillae. In addition, a list of species assigned to Cosmocercoides is provided; however, C. fotedari Arya, 1992 is removed from the genus and until further study is considered a species inquirenda. Keywords Nematoda, Cosmocercoides himalayanus sp. nov., Anura, Duttaphrynus himalayanus, India Introduction During a faunistic survey of Dehardun, India, 3 specimens of the Himalayan toad Duttaphrynus himalayanus (Gunther, 1864) were collected. At necropsy, nematodes assignable to the genus Cosmocercoides Wilkie, 1930 were found. Duttaphrynus himalayanus (previously Bufo himalayanus) is known from southern slopes of the Himalayas in China, Pakistan, Nepal, India, and Bangladesh at elevations of 2000 3500 m; presumably also in adjacent Bhutan and Myanmar (Frost 2013). It has been previously reported to harbor 4 species of nematodes, Cosmocercoides bufonis, Meteterakis govindi, Oswaldocruzia filiformis and Rhabdias himalayanus (Karve 1944; Soota and Dey Sarkar 1980; Rizvi et al. 2013). The genus Cosmocercoides is currently comprised of 20 species (Table I). The purpose of this paper is to describe the 21st species assigned to the genus, Cosmocercoides himalayanus sp. nov. Materials and Methods The toads were brought live to the laboratory and pithed. The body cavity was opened by a longitudinal lateral incision and the gastrointestinal tract was removed. The oesophagus, stomach, small intestine, large intestine and rectum of each host were examined separately for helminths. Only nematodes were found which were fixed in formalin, dehydrated in glycerin alcohol and examined in wax sealed slides. They were subsequently photomicrographed by BX51 DIC/BF Olympus research microscope with DP20 digital camera. Measurements are in µm unless otherwise noted and given as range with holotype or allotype in parentheses. Results Cosmocercoides himalayanus sp. nov. (Figs 1 9) General morphology: Small, stout nematodes. Sexual dimorphism evident, males two-thirds length of females. Lateral alae present in both males and females. Cuticle transversely striated. Somatic papillae present in both males and females. Mouth with three prominent lips, dorsal lip with two sessile papillae, each ventrolateral lip with one ventral sessile papilla and one lateral amphid. Shallow mouth cavity, short pharynx, cylindrical corpus, short isthmus and valved bulb present. Excretory pore anterior to oesophageal bulb. *Corresponding author: cxb13@psu.edu

New species of Cosmocercoides 81 Table I. Species assigned to Cosmocercoides Cosmocercoides Wilkie, 1930 Syn. Trionchonema Kreis, 1932 [Chitwood 1933] C. barodensis Rao, 1979 [India; toad host, Bufo melanostictus (currently, Duttaphrynus melanostictus)] C. bufonis Karve, 1944 [India; toad host Bufo himalayanus (currently, Duttaphrynus himalayanus)] C. dukae (Holl, 1928) Travassos, 1931 [USA, North Carolina; host Triturus viridescens (currently Notophthalmus viridescens)] Syn. Cosmocerca dukae Holl, 1928 Syn. Aplectana americana Walton, 1929 [Baker 1981] Syn. Trichonema rusticum Kreis, 1932 Syn. Cosmocercoides rusticum (Kreis, 1932) Chitwood 1933 [Baker, 1987] Syn. Oxysomatium georgianun Reiber, Byrd & Parker, 1940 [Baker 1980] Syn. Aplectana lynae Kennedy, 1977 [Baker 1981] C. himalayanus sp. nov. [India; toad host, Duttaphrynus himalayanus] C. karnatakaensis Rizvi, 2009 [India; frog host, Hylarana temporalis] C. kumaoni Arya, 1992 [India; frog host, Rana cyanophlyctis (currently, Euphlyctis cyanophlyctis)] C. lanceolatus Rao, 1979 [India; frog host, Rana cyanophlyctis (currently, Euphlyctis cyanophlyctis)] C. lilloi Ramallo, Bursey and Goldberg, 2007 [Argentina; toad host, Chaunus arenarium (currently Rhinella arenarum)] C. microhylae (Wang, Zhao and Chen, 1978) Baker, 1987 [China; host Microhyla ornata] Syn. Paracosmocerca microhylae Wang, Zhao and Chen, 1978 C. multipapillata Khera, 1958 [India; toad host, Bufo melanostictus (currently, Duttaphrynus melanostictus)] C. nainitalensis Arya, 1979 [India: frog host, Rana cyanophlyctis (currently, Euphlyctis cyanophlyctis)] C. oligodentis Wang, Sun, Zhao and Zhang, 1981 [China; host Oligodon chinensis] C. pulcher Wilkie, 1930 [Japan; host Rana japonica] TYPE SPECIES C. ranae Wang, Sun, Zhao and Zhang, 1981 [China; host Rana spinosa (currently Quasipaa spinosa] C. rickae Ogden, 1966 [Sri Lanka; lizard host, Ceratophora tennenti] C. sauria Avila, Strussmann and Silva, 2010 [Brazil; lizard host, Iphisa elegans] C. skrjabini (Ivanitsky, 1940) Skrjabin and Shikhobalova, 1951 [Ukraine; frog host, Rana temporaria]] Syn. Cosmocerca skrjabini Ivanitsky, 1940 C. speleomantis Ricci, 1987 [Italy; host Speleomantes flavus (currently Hydromantoides flavus)] C. tibetanum (Baylis, 1927) Baker 1980 [Tibet; host Scutiger alticola (currently Scutiger boulengeri)] Syn. Oxysomatium tibetanum Baylis, 1927 C. tridens Wilkie, 1930 [Japan; Redescription: Hasegawa 1989; new host, Tylototriton andersoni (currently Echinotriton andersoni)] C. variabilis (Harwood, 1930) Travassos 1931 [USA, Texas; toad host, Bufo valliceps (currently Incilius valliceps)] Syn. Cosmocerca variabilis Harwood, 1930 Male (holotype and 14 paratypes): Length 4.56 6.77 (5.78) mm; width at midbody 300 470 (407). Oesophagus including bulb 910 1020 (958), buccal cavity 10 12 (10), pharynx 75 82 (77), corpus 750 830 (800), isthmus 40 50 (46), bulb length 120 140 (135), bulb width 125 140 (130). Nerve ring 400 410 (406); excretory pore 520 600 (560) from anterior end. Tail 200 250 (230) long, tapering. Alate spicules similar, 242 260 (250) long; gubernaculum 142 158 (148) in length. Twenty one pairs of caudal papillae: 15 pairs of rosette papillae, 12 pairs precloacal, 1 pair adcloacal, 2 pairs postcloacal, all ventrolateral in position; 6 pairs of simple papillae, all postcloacal in position, 4 pairs ventrolateral, 2 pairs dorsolateral; phasmids located midway between dorsolateral pairs of postcloacal papillae. Female (allotype and 9 paratypes): Length 6.43 7.70 (6.90) mm; width at midbody 340 450 (400). Oesophagus including bulb 860 1200 (1100), buccal cavity 10 12 (10), pharynx 75 82 (80), corpus 660 1000 (850), isthmus 50 60 (56) bulb length 130 150 (135), bulb width 120 140 (130). Nerve ring 370 422 (390); excretory pore 450 650 (630) from anterior end. Vulva 3.65 4.41 (3.90) mm from anterior end (57% of body length from anterior end). Short muscular ovijector giving way to thin walled vagina, which joins 2 uteri. Tail 230 300 (280) in length. Eggs, oval, thin-shelled and in different stages of development, 50 60 37 40. Taxonomic summary Type host: Himalayan toad, Duttaphrynus himalayanus (Günther, 1864); symbiotype, ZSI/NRC/FC7. Type locality: Branskhanda, Dehrandun, Uttarakhand, India, 30 26.547 N, 78 12.392 E; 2372 m elevation. Site of infection: Large intestine. Prevalence of infection: 2 of 3 hosts (66%); 10 females and 15 males collected. Type material: Holotype male, ZSI/NRC/IV.1027; paratype males, ZSI/NRC/IV.1028; allotype female, ZSI/NRC/IV.1028; paratype females, ZSI/NRC/IV.1029. Etymology: The new species is named after the host species, Duttaphrynus himalayanus. Remarks Cosmocercoides himalayanus sp. nov. is the 21st species assigned to the genus and the 9th species described from the Oriental region. Table I lists the current species assigned to Cosmocercoides. It should be noted that we consider Cosmocercoides fotedari Arya, 1992 from Euphlyctis cyanophlyctis (previously Rana cyanophlyctis) of India to be a species inquirenda; it lacks rosette papillae and the oesophagus is

82 Anjum N. Rizvi and Charles R. Bursey similar to that seen in the genus Falcaustra but further study is required before assignment can be made. Oriental species of Cosmocercoides, with the exception of C. rickae (a parasite of the lizard, Ceratophora tennenti), are known only from frogs and toads; Palaearctic and Western Hemisphere species are known from frogs, salamanders, lizards and snakes. The life cycle of the new species is unknown, but studies on C. variabilis in anurans have shown that Figs 1 9. Cosmocercoides himalayanus sp. nov. 1. Female, anterior end, lateral view. 2. En face view. 3. Egg. 4. Caudal rosette. 5. Male, posterior end, lateral view. 6. Female, posterior end, lateral view. 7. Gubernaculum. 8. Spicule. 9. Male, posterior end, ventral view

New species of Cosmocercoides 83 Table II. Selected characters of male individuals of species of Cosmocercoides Reference Lateral alae Prebulbar swelling Rosette papillae* Gubernaculum Spicule Tail Length (mm) Biogeographical region Cosmocercoides spp. Nearctic C. dukae 2.6 240 Equal: 290 102 22:0:2 0 Not stated Holl, 1928 C. variabilis 2.2 5.0 125 220 Equal: 250 420 110 160 24 36:2:6 0 Present Harwood, 1930 Neotropical C. lilloi 2.0 2.4 160 210 Equal:100 120 Absent 18 19:0:8 +2 0 Absent Ramallo et al., 2007 C. sauria 1.3 54 Equal:104 97 8:0:0 0 Present Avila et al., 2010 Oriental C. himalayanus sp. nov. 4.6 6.8 200 250 Equal: 242 260 142 158 24:2:4 0 Present Present study C. barodensis 2.1 200 Subequal: 240 & 230 60 16:0:4 0 Present Rao, 1979 C. bufonis 4.6 4.9 200 240 Equal: 190 260 120 145 18 26:2:6 1 Present Karve, 1944 C. karnatakaensis 1.3 1.6 250 290 Equal: 245 265 31 42 16:10:24 0 Absent Rizvi, 2009 absent 24:2:10 0 Present Arya, 1991 Subequal: 100 160 & 90 125 C. kumaoni 2.0 3.0 130 150 C. lanceolaltus 1.4 250 Equal: 210 76 10:2:2 0 Present Rao, 1979 C. multipapillata 5.8 6.4 250 280 Equal: 200 240 130 140 21 27:2:8 1 Present Khera, 1958 100 120 26 32:2:12 2 Present Arya, 1979 Subequal: 130 151 & 112 134 C. nainitalensis 4.9 5.5 230 250 C. rickae 3.1 161 Equal: 168 121 28:2:6 0 Present Ogden, 1966 Palaearctic C. microhylae 2.2 157 Equal: 140 Absent 20:0:0 0 Present Wang et al., 1978 C. oligodentis 2.6 320 Equal: 112 45 16:0:0 0 Present Wang et al., 1981 C. pulcher 6.9 220 Equal: 247 143 28 30:4:2 0 Absent Wilkie, 1930 C. ranae 2.4 302 Equal: 192 105 20:0:0 0 Present Wang et al., 1981 C. skrjabini Equal: 212 210 16:0:0 0 Present Ivanitzky, 1940 C. speleomantis 1.8 2.3 115 119 Not seen 81 88 8:0:0 0 Present Ricci, 1987 C. tibetanum 4.5 5.0 170 Equal: 580 188 44:0:2 0 Absent Baylis, 1927 C. tridens 2.7 5.0 140 210 Equal: 270 530 96 146 22 30:0:6 0 Present Hasegawa, 1989 *precloacal:adcloacal:postcloacal + median

84 Anjum N. Rizvi and Charles R. Bursey released eggs develop into first stage larvae in the external environment and molt twice to the infective third stage larvae before infecting a new host by skin penetration (Anderson 2000). Selected characters of males of species of Cosmocercoides are presented in Table II. Of the species listed in Table II, only C. bufonis, C. kumoani, C. multipapillata, C. nainitalensis, C. pulcher, C. rickae, C. tibetanum, C. tridens, C. variabilis and the new species have 24 or more rosette (12 or more pairs) papillae. Of these, only the new species has 4 postcloacal (2 pairs) rosette papillae; C. pulcher and C. tibetanum possess 2 of postcloacal rosette papillae; C. bufonis, C. rickae, C. tridens and C. variabilis each possess 6; C. multipapillata, 8; C. kumaoni, 10 and C. nainitalensis, 12. Acknowledgements. Our thanks to the Director, Dr K. Venkataraman, Zoological Survey of India (ZSI), Kolkata and the Officer-in- Charge, ZSI, Northern Regional Centre, for facilities and encouragements. We thank Dr P.C. Tak and the members of the survey team from ZSI, Dehradun for collection of amphibian hosts and Shri. C. Radhakrishnan, Western Ghat Regional Centre, ZSI, Kozhikode, for identification of the hosts. References Anderson R.C. 2000. Nematode parasites of vertebrates. Their development and transmission. 2st ed. CABI Publishing, Wallingford, U.K., 650 pp. Arya S.N. 1979. A new nematode, Cosmocercoides nainitalensis n. sp. from the frog, Rana cyanophlyctis. Revista Brasileira Biologia, 39, 823 825. Arya S.N. 1991. Two new species of the genus Cosmocercoides Wilkie, 1930 from a frog Rana cyanophlyctis from Nainital. Indian Journal of Helminthology, 43, 139 143. Avila R.W., C. Strussmann, and R.J. da Silva. 2010. A new species of Cosmocercoides (Nematoda: Cosmocercidae) from a gymnophthalmid lizard of western Brazil. Journal of Parasitology, 96, 558 560. DOI: 10.1645/GE-2336.1. Baylis H.A. 1927. On two new species of Oxysomatium (Nematoda) with some remarks on the genus. Annals and Magazine of Natural History, Series 9, 19, 279 286. Frost D.R. 2013. Amphibian species of the world: an online reference. Version 5.6 (9 January 2013). Electronic database accessible at http://research.amnh.org/herpetology/amphibia/ index.html. American Museum of Natural History, New York, USA. Accessed 22 May 2013. Harwood P.D. 1930. The new species of Oxysomatium (Nematoda) with some remarks on the genera Oxysomatium and Aplectana and observations on the life history. Journal of Parasitology, 17, 61 73. DOI: 10.2307/3271435. Hasegawa H. 1989. Redescription of Cosmocercoides tridens Wilkie, 1930 (Nematoda: Cosmocercidae). Canadian Journal of Zoology, 67, 1189 1193. DOI: 10.1139/z89-171. Holl F.J. 1928. Two new nematode parasites. Journal of the Elisha Mitchell Scientific Society, 43, 184 186. Ivanitzky S.V. 1940. [On the helminth fauna of vertebrates in the Ukraine (Cestoda, Nematoda and Acanthocephala).] Sbornik Trudy Khar kov Veterinary Institute, 19, 129 155 (In Russian). Karve J.N. 1944. On a small collection of parasitic nematodes from Anura. Proceedings of the Indian Academy of Science, 20, 71 77. Khera S. 1958. On a new species of Cosmocercoides. Indian Journal of Helminthology, 10, 6 12. Ogden C.G. 1966. On some parasitic nematodes from reptiles, mainly from Ceylon. Journal of Helminthology, 15, 81 90. DOI: 10.1017/S0022149X00034118. Ramallo G., C.R. Bursey and S.R. Goldberg. 2007. Two new species of cosmocercids (Ascaridida) in the toad Chaunus arenarum (Anura: Bufonidae) from Argentina. Journal of Parasitology, 93, 910 916. DOI: 10.1645/GE-1131R.1. Rao R. 1979. On four new species of the genera Paracosmocerca Kung et Wu, 1945, Cosmocerca Diesing, 1861 and Cosmocercoides Wilkie, 1930 of the family Cosmocercidae Travassos, 1925. Helminthologia, 16, 23 34. Ricci M. 1987. Parasites of Speleomantes (Amphibia: Urodela: Plethotidae) in Italy. Rivista di Parassitologia, 4, 5 25. Rizvi A.N. 2009. Two new species of amphibian nematodes from Bhadra Wildlife Sanctuary, Western Ghats, India. Zootaxa, 2013, 58 68. Rizvi A.N., C.R. Bursey and P.T. Bhutia. 2013. Two new species of Rhabdias (Nematoda: Rhabditida: Rhabdiasidae) in anuran hosts from Dehradun (Uttarkhand), India. Journal of Parasitology, 99, 303 306. DOI: 10.1645/GE-3179-1. Soota T.D., and S.R. Dey Sarkar. 1980. On some nematodes from Darjeeling District, West Bengal, India. Proceedings of the Zoological Society of Calcutta, 31, 39 46. Wang P. Q. Y. L. Sun, Y. R. Zhao, and W. H. Zhang. 1981. Notes on five new species of nematodes from vertebrates in Wuyi, Fujian Province. Wuyi Science Journal, 1, 113 118. Wang P.Q., Y. Zhao, and C. Chen. 1978. On some nematodes from vertebrates in south China. Fjuian Shida Xuebao, 2, 75 90. Wilkie J. S. 1930. Some parasitic nematodes from Japanese Amphibia. Annals and Magazine of Natural History, Series 10, 6: 606 614. Received: June 23, 2012 Revised: August 12, 2013 Accepted for publication: December 18, 2013