DOI: 10.2478/s11686-014-0212-8 © W. Stefański Institute of Parasitology, PAS Acta Parasitologica, 2014, 59(1), 80–84; ISSN 1230-2821 himalayanus sp. nov. (Nematoda, ) in himalayanus (Amphibia, Anura) from Dehradun (Uttarakhand), India

Anjum N. Rizvi1 and Charles R. Bursey2* 1Zoological Survey of India (ZSI), Northern Regional Centre, Dehradun-248 195, Uttarakhand, India; 2Department of Biology, Pennsylvania State University, Shenango Campus, Sharon, Pennsylvania 16146, USA

Abstract Cosmocercoides himalayanus sp. nov. (Nematoda, Cosmocercidae) from the large intestine of (Amphibia, Anura) from Dehradun, India is described and illustrated. Cosmocercoides himalayanus sp. nov. represents the 21st species assigned to the 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 pos- sessing 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 the gastrointestinal tract was removed. The oesophagus, stom- ach, small intestine, large intestine and rectum of each host During a faunistic survey of Dehardun, India, 3 specimens were examined separately for helminths. Only of the Himalayan Duttaphrynus himalayanus (Gunther, were found which were fixed in formalin, dehydrated in 1864) were collected. At necropsy, nematodes assignable glycerin alcohol and examined in wax sealed slides. They to the genus Cosmocercoides Wilkie, 1930 were found. were subsequently photomicrographed by BX51 DIC/BF Duttaphrynus himalayanus (previously himalayanus) Olympus research microscope with DP20 digital camera. is known from southern slopes of the Himalayas in China, Measurements are in µm unless otherwise noted and given as Pakistan, Nepal, India, and Bangladesh at elevations of range with holotype or allotype in parentheses. 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, Results Meteterakis govindi, Oswaldocruzia filiformis and Rhabdias himalayanus (Karve 1944; Soota and Dey Sarkar 1980; Cosmocercoides himalayanus sp. nov. (Figs 1–9) Rizvi et al. 2013). The genus Cosmocercoides is currently comprised of 20 species (Table I). The purpose of this General morphology: Small, stout nematodes. Sexual dimor- paper is to describe the 21st species assigned to the genus, phism evident, males two-thirds length of females. Lateral alae Cosmocercoides himalayanus sp. nov. present in both males and females. Cuticle transversely stri- ated. Somatic papillae present in both males and females. Mouth with three prominent lips, dorsal lip with two sessile Materials and Methods papillae, each ventrolateral lip with one ventral sessile papilla and one lateral amphid. Shallow mouth cavity, short pharynx, The were brought live to the laboratory and pithed. The cylindrical corpus, short isthmus and valved bulb present. body cavity was opened by a longitudinal lateral incision and Excretory pore anterior to oesophageal bulb.

*Corresponding author: [email protected] 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. 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; 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) Taxonomic summary mm; width at midbody 300–470 (407). Oesophagus including bulb 910–1020 (958), buccal cavity 10–12 (10), pharynx 75–82 Type host: Himalayan toad, Duttaphrynus himalayanus (Gün- (77), corpus 750–830 (800), isthmus 40–50 (46), bulb length ther, 1864); symbiotype, ZSI/NRC/FC7. 120–140 (135), bulb width 125–140 (130). Nerve ring 400–410 Type locality: Branskhanda, Dehrandun, Uttarakhand, (406); excretory pore 520–600 (560) from anterior end. Tail 200– India, 30°26.547´N, 78°12.392´E; 2372 m elevation. 250 (230) long, tapering. Alate spicules similar, 242–260 (250) Site of infection: Large intestine. long; gubernaculum 142–158 (148) in length. Twenty one pairs Prevalence of infection: 2 of 3 hosts (66%); 10 females of caudal papillae: 15 pairs of rosette papillae, 12 pairs precloa- and 15 males collected. cal, 1 pair adcloacal, 2 pairs postcloacal, all ventrolateral in Type material: Holotype male, ZSI/NRC/IV.1027; paratype position; 6 pairs of simple papillae, all postcloacal in position, males, ZSI/NRC/IV.1028; allotype female, ZSI/NRC/IV.1028; 4 pairs ventrolateral, 2 pairs dorsolateral; phasmids located paratype females, ZSI/NRC/IV.1029. midway between dorsolateral pairs of postcloacal papillae. Etymology: The new species is named after the host Female (allotype and 9 paratypes): Length 6.43–7.70 species, Duttaphrynus himalayanus. (6.90) mm; width at midbody 340–450 (400). Oesophagus in- cluding bulb 860–1200 (1100), buccal cavity 10–12 (10), Remarks pharynx 75–82 (80), corpus 660–1000 (850), isthmus 50–60 (56) bulb length 130–150 (135), bulb width 120–140 (130). Cosmocercoides himalayanus sp. nov. is the 21st species as- Nerve ring 370–422 (390); excretory pore 450–650 (630) from signed to the genus and the 9th species described from the anterior end. Vulva 3.65–4.41 (3.90) mm from anterior end Oriental region. Table I lists the current species assigned to (57% of body length from anterior end). Short muscular ovi- Cosmocercoides. It should be noted that we consider Cosmo- jector giving way to thin walled vagina, which joins 2 uteri. cercoides fotedari Arya, 1992 from Euphlyctis cyanophlyctis Tail 230–300 (280) in length. Eggs, oval, thin-shelled and in (previously Rana cyanophlyctis) of India to be a species different stages of development, 50–60 × 37–40. 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 known only from and toads; Palaearctic and Western is required before assignment can be made. Hemisphere species are known from frogs, salamanders, Oriental species of Cosmocercoides, with the exception of lizards and snakes. The life cycle of the new species is un- C. rickae (a parasite of the lizard, Ceratophora tennenti), are known, 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 ., 2007 ., 1978 ., 1981 ., 1981 ., 2010 et al et al et al et al et al Rao, 1979 Rao, 1979 Reference Holl, 1928 Arya, 1991 Arya, 1979 Ricci, 1987 Rizvi, 2009 Khera, 1958 Karve, 1944 Baylis, 1927 Ogden, 1966 Wilkie, 1930 Wilkie, Present study Ivanitzky, 1940 Ivanitzky, Harwood, 1930 Hasegawa, 1989 Avila Avila Wang Wang Wang Wang Wang Ramallo alae Absent Absent Absent Absent Present Present Present Present Present Present Present Present Present Present Present Present Present Present Present Present Lateral Not stated 1 2 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 swelling Prebulbar 8:0:0 8:0:0 28:2:6 20:0:0 16:0:0 16:0:0 22:0:2 10:2:2 20:0:0 24:2:4 16:0:4 44:0:2 24:2:10 Rosette 16:10:24 papillae* 18–26:2:6 21–27:2:8 24–36:2:6 28–30:4:2 22–30:0:6 26–32:2:12 18–19:0:8 +2 97 45 60 76 143 210 102 121 105 188 (um) 81–88 31–42 absent Absent Absent 96–146 110–160 100–120 142–158 120–145 130–140 Gubernaculum (um) Spicule Not seen & 90–125 Equal:104 Equal: 112 Equal: 247 Equal: 192 Equal: 290 Equal: 210 Equal: 168 Equal: 140 Equal: 212 Equal: 580 & 112–134 Equal:100–120 Equal: 270–530 Equal: 242–260 Equal: 190–260 Equal: 245–265 Equal: 200–240 Equal: 250–420 Subequal: 100–160 Subequal: 130–151 Subequal: 240 & 230 Cosmocercoides – 54 220 240 157 320 170 200 250 161 302 Tail (um) 115–119 125–220 160–210 200–240 200–250 250–290 130–150 230–250 140–210 250–280 – 6.9 2.6 1.3 2.2 2.6 2.1 3.1 2.4 1.4 (mm) 2.2–5.0 2.0–2.4 4.6–4.9 1.3–1.6 2.0–3.0 1.8–2.3 4.6–6.8 4.9–5.5 2.7–5.0 5.8–6.4 4.5–5.0 Length spp. sp. nov. Selected characters of male individuals species C. himalayanus C. dukae C. lilloi C. sauria C. nainitalensis C. rickae C. ranae C. variabilis C. karnatakaensis C. kumaoni C. lanceolaltus C. multipapillata C. microhylae C. oligodentis C. pulcher C. speleomantis C. tibetanum C. tridens C. barodensis C. bufonis C. skrjabini Cosmocercoides Biogeographical region Table II. Table Nearctic Oriental Neotropical Palaearctic *precloacal:adcloacal:postcloacal + median 84 Anjum N. Rizvi and Charles R. Bursey

released eggs develop into first stage larvae in the external en- Harwood P.D. 1930. The new species of Oxysomatium (Nematoda) vironment and molt twice to the infective third stage larvae be- with some remarks on the genera Oxysomatium and Aplectana and observations on the life history. Journal of Parasitology, fore infecting a new host by skin penetration (Anderson 2000). 17, 61–73. DOI: 10.2307/3271435. Selected characters of males of species of Cosmocercoides Hasegawa H. 1989. Redescription of Cosmocercoides tridens Wilkie, are presented in Table II. Of the species listed in Table II, only 1930 (Nematoda: Cosmocercidae). Canadian Journal of Zo- C. bufonis, C. kumoani, C. multipapillata, C. nainitalensis, ology, 67, 1189–1193. DOI: 10.1139/z89-171. C. pulcher, C. rickae, C. tibetanum, C. tridens, C. variabilis Holl F.J. 1928. Two new parasites. Journal of the Elisha Mitchell Scientific Society, 43, 184–186. and the new species have 24 or more rosette (12 or more pairs) Ivanitzky S.V. 1940. [On the helminth fauna of vertebrates in the papillae. Of these, only the new species has 4 postcloacal Ukraine (Cestoda, Nematoda and Acanthocephala).] (2 pairs) rosette papillae; C. pulcher and C. tibetanum possess Sbornik Trudy Khar’kov Veterinary Institute, 19, 129–155 2 of postcloacal rosette papillae; C. bufonis, C. rickae, (In Russian). C. tridens and C. variabilis each possess 6; C. multipapillata, Karve J.N. 1944. On a small collection of parasitic nematodes from Anura. Proceedings of the Indian Academy of Science, 8; C. kumaoni, 10 and C. nainitalensis, 12. 20, 71–77. Khera S. 1958. On a new species of Cosmocercoides. Indian Journal Acknowledgements. Our thanks to the Director, Dr K. Venkatara- of Helminthology, 10, 6–12. man, Zoological Survey of India (ZSI), Kolkata and the Officer-in- Ogden C.G. 1966. On some parasitic nematodes from reptiles, mainly Charge, ZSI, Northern Regional Centre, for facilities and from Ceylon. Journal of Helminthology, 15, 81–90. DOI: encouragements. We thank Dr P.C. Tak and the members of the sur- 10.1017/S0022149X00034118. vey team from ZSI, Dehradun for collection of hosts and Ramallo G., C.R. Bursey and S.R. Goldberg. 2007. Two new species Shri. C. Radhakrishnan, Western Ghat Regional Centre, ZSI, of cosmocercids () in the toad Chaunus arenarum Kozhikode, for identification of the hosts. (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 Cosmo- References cercoides Wilkie, 1930 of the family Cosmocercidae Travas- sos, 1925. Helminthologia, 16, 23–34. Anderson R.C. 2000. Nematode parasites of vertebrates. Their de- Ricci M. 1987. Parasites of Speleomantes (Amphibia: Urodela: velopment and transmission. 2st ed. CABI Publishing, Plethotidae) in Italy. Rivista di Parassitologia, 4, 5–25. Wallingford, U.K., 650 pp. Rizvi A.N. 2009. Two new species of amphibian nematodes from Arya S.N. 1979. A new nematode, Cosmocercoides nainitalensis n. Bhadra Wildlife Sanctuary, Western Ghats, India. Zootaxa, sp. from the frog, Rana cyanophlyctis. Revista Brasileira Bi- 2013, 58–68. ologia, 39, 823–825. Rizvi A.N., C.R. Bursey and P.T. Bhutia. 2013. Two new species Arya S.N. 1991. Two new species of the genus Cosmocercoides of Rhabdias (Nematoda: Rhabditida: Rhabdiasidae) Wilkie, 1930 from a frog Rana cyanophlyctis from Nainital. in anuran hosts from Dehradun (Uttarkhand), India. Journal Indian Journal of Helminthology, 43, 139–143. of Parasitology, 99, 303–306. DOI: 10.1645/GE-3179-1. Avila R.W., C. Strussmann, and R.J. da Silva. 2010. A new species of Soota T.D., and S.R. Dey Sarkar. 1980. On some nematodes from Cosmocercoides (Nematoda: Cosmocercidae) from a Darjeeling District, West Bengal, India. Proceedings of the gymnophthalmid lizard of western Brazil. Journal of Para- Zoological Society of Calcutta, 31, 39–46. sitology, 96, 558–560. DOI: 10.1645/GE-2336.1. Wang P. Q. Y. L. Sun, Y. R. Zhao, and W. H. Zhang. 1981. Notes on Baylis H.A. 1927. On two new species of Oxysomatium (Nematoda) five new species of nematodes from vertebrates in Wuyi, Fu- with some remarks on the genus. Annals and Magazine of jian Province. Wuyi Science Journal, 1, 113–118. Natural History, Series 9, 19, 279–286. Wang P.Q., Y. Zhao, and C. Chen. 1978. On some nematodes from Frost D.R. 2013. Amphibian species of the world: an online refer- vertebrates in south China. Fjuian Shida Xuebao, 2, 75–90. ence. Version 5.6 (9 January 2013). Electronic database ac- Wilkie J. S. 1930. Some parasitic nematodes from Japanese Am- cessible at http://research.amnh.org/herpetology/amphibia/ phibia. Annals and Magazine of Natural History, Series 10, 6: index.html. American Museum of Natural History, New York, 606–614. USA. Accessed 22 May 2013.

Received: June 23, 2012 Revised: August 12, 2013 Accepted for publication: December 18, 2013