Two New Trematodes of Family Acanthocolpidae Luhe, 1906 From Marine Fish Leiognathus daura (Cuvier) from the Coast of Puri, Orissa, India

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Iranian J Parasitol Tehran University of Medical Sciences Publication http:// tums.ac.ir Open access Journal at http:// ijpa.tums.ac.ir Iranian Society of Parasitology http:// isp.tums.ac.ir Case Report Two New Trematodes of Family Acanthocolpidae Luhe, 1906 From Marine Fish Leiognathus daura (Cuvier) from the Coast of Puri, Orissa, India Sushma MISHRA, Satish CHANDRA, *Anand Murari SAXENA Department of Zoology, University of Lucknow, Lucknow, India Received 28 Mar 2013 Accepted 12 July 2013 Keywords Digenea, Marine fishes, Acanthocolpus, Parasite, India *Correspondence Email: anandmsaxena@rediffmail.com Abstract Background: Genus Acanthocolpus (Trematoda: Acanthocolpiidae) is one of the most important zoonotic digenean with wide geographic distribution in the world. The purpose of the present study was to describe morphological and morphometrical characteristics of Acanthocolpus species, currently prevalent in marine fish fauna of Puri coast, Orissa, India. Methods: Gastro-intestinal organs of Leiognathus daura (Cuvier) in Puri coast, Orissa, India, were examined for infectivity with digenean trematode species. For examination and measurements of helminthes, acetoalum carmine staining was performed, followed by camera Lucida drawings of morphological characters and measurements of morphometrical criteria with a calibrated microscope. Using valid trematode systematic keys, almost all the parasites were identified at the level of species. Results: Overall, 36 marine fishes were found infected with at least one species of Acanthocolpus. Considering morphological characteristics of Acanthocolpus, two species were identified as new species including Acanthocolpus durghai sp.nov. and Acanthocolpus amrawatai sp.nov. Conclusion: During the survey of helminth parasites, collected six different species of the genus Acanthocolpus, out of these two are new species, another is redescribed to show certain variation, the new parasite was obtain from the intestine of marine fish Leiognathus daura (Cuvier) 494 Available at: http://ijpa.tums.ac.ir

Mishr et al.: Two New Trematodes of Family Acanthocolpidae Luhe Introduction I n this paper we are adding the knowledge of Acanthocolpus (1), especially of marine teleost fishes from the coast of Puri, Orissa (India). During the survey of helminth parasites, collected six different species of the genus Acanthocolpus, are found out of these two are new and four species, are rediscribed. The new species were found in the intestine of marine fish Leiognathus daura (Cuvier). intestinal caeca is simple extending up to hind end of body. Ventral sucker is present at anterior extremity, sub-spherical, pre-testicular, larger than oral sucker, 0.11-0.13 mm long, 0.12-0.14 mm wide at 0.10-0.20 mm from anterior extremity. Materials and Methods During the examination of the marine fish specimens of the above genus were recovered from the intestine of marine fish. The specimens were collected and identified by fish books and cut open and thoroughly examined after that helminthes parasites were separated in Petri dish containing normal saline solution. The parasite were flattened with slight pressure of cover glass and fixed in A.F.A. fixative (50% alcohol, formalin and acetic acid in ratio of 100: 6: 2.5). They were stained in acetoalum carmine, differentiated in acid alcohol and dehydrated through ascending grade of alcohols. These were cleared in xylol and mounted in canada balsam or DPX. The diagrams were made with the help of camera Lucida. All the measurements in millimeters: unless otherwise stated. Results Description Family: Acanthocolpiidae Subfabily: Acanthocolpiinae Acanthocolpus durghai sp nov. (Figure-1) Body is elongated, aspinose, slender 1.69-1.86 mm long, 0.28-0.30 mm wide. Oral sucker Reni form terminal, 0.05-0.08 mm long, 0.08-0.11 mm wide. Pre-pharynx Long, narrow, 0.04-0.11 mm long, 0.01-0.01mm wide. Pharynx small ovoid muscular 0.03-0.04 mm long 0.02-0.02 mm wide. Oesophagus absent, Fig.1: Acanthocolpus durghai sp. nov.; 1.1 Ventral view of adult; 1.2 Eggs Excretory bladder Y -shaped, excretory pore terminal. Genital pore sub-median, lying on right side of body behind oral sucker at 0.05-0.13 mm from anterior extremity. Testes two, elongated, inter-caecal, in posterior third of the body. Anterior testis 0.19-0.22 mm long, 0.11-0.13 mm wide at 0.86-1.14 mm from anterior extremity. Posterior testis 0.09-0.30 mm long, 0.10-0.12 mm wide at 1.07-1.36 mm from anterior extremity. Cirrus sac long, slender, sinuous, 0.56-1.01 mm long, 0.08-0.09 Available at: http://ijpa.tums.ac.ir 495

mm wide extending from just in front of ovary up to little behind to oral sucker. Vesicula seminalis elongated, 0.32-0.53 mm long, 0.06-0.07 mm wide. Pars-prostatica long, tubular, 0.25-0.49 mm long surrounded by large number of prostate gland cells. Ovary sub-spherical, sub-median, intercaecal, pre-testicular, post-equatorial just in front of anterior testis, 0.11-0.12 mm long, 0.09-0.10 mm wide at 0.75-1.07 mm from anterior extremity. Vitelleria follicle extending from middle of cirrus sac up to hind end of body. Uterus pre-ovarian. Metaterm muscular. Eggs large, elongated, operculated, 0.03-0.04 mm long, 0.01-0.02 mm wide. Discussion The present form belongs to genus Acanthocolpus Luhe, 1906 comprises of the following species A. liodorus (1) from Ceylon Synonym A. luhei (2); A. indicum (2) from Karachi; A. orientalis (2) from Karachi; A. tenuis (3) from Fije, A. brasilensis (4) from Guanabara, A. caballeroi (5) from Ratna Giri; A. guptai, A. puriensis, A. lucknowensis (6) from Puri, Orissa, A. inglisi (6) from Puri, Orissa, A. srivastavai, A. valiyathurai (7) from Trivandrum, Kerala, A. chorinesmusi, A. equulai, A. lutijanusi (8) from Tamilnadu and Puri, Orissa; A. microtesticulus (9) from Maharashtra are known so far. A. luhei was considered as a synonym of A. liodorus (10). A. luhei was distinguished from A. liodorus in the anterior extent of vitellaria, sucker ratio and in the absence of acetabular penduncle (11). Later on Gupta and Gupta (12) followed Yamaguti (10) and considered all the characters mentioned by Hafeezullah, (11) are variable characters. Author agrees with Yamaguti (10) and Gupta and Gupta (12) in considering A. luhei (2) as a synonym of A. liodorus (1). The present form differs from all these species except A. luhei, A. indicum, A. orientalis and A. chorinesmusi in the absence of acetabular peduncle. The present form differs from all these forms in having reniform oral sucker instead of sub-spherical or conical, genital pore lying on little posterior to oral sucker instead of just pre-acetabular region or at middle of ventral sucker, cirrus sac extending from just anterior to ovary instead of separated from uterine coils and in relative shape and size of various organ. Description Acanthocolpus amrawatai sp. nov. (Figure-2) Body elongated, aspinose, slender, 1.91-3.18 mm long, 0.27-0.37 mm wide. Oral sucker sub-terminal, sub-spherical, 0.11-0.12 mm long, 0.09-0.12 mm wide. Pre-pharynx long, narrow, tubular, 0.04 0.13 mm long, 0.02 mm wide. Pharynx ovoid, muscular 0.08-0.09 mm long, 0.03-0.03 mm wide. Oesophagous very short. Intestinal caeca simple, extending up to near posterior extremity. Ventral sucker larger than oral sucker, sub-spherical, pedunculated, 0.11-0.15 mm long, 0.15-0.16 mm wide at 0.11-0.36 mm from anterior extremity. Excretory bladder Y - shaped, excretory pore terminal. Genital pore lying posterior to ventral sucker at 0.21-0.37 mm from anterior extremity. Testes two, lying one behind other, inter-caecal, post-ovarian, in posterior third of the body. Anterior testis pear shaped 0.23-0.29 mm long, 0.12-0.13 mm wide at 1.09-1.94 mm from anterior extremity. Posterior testis elongated 0.24-0.34 mm long, 0.12-0.12 mm wide at 1.37-2.33 mm from anterior extremity. Cirrus sac long, elongated, tubular, sinuous, 0.84-1.53 mm long, extending from anterio-lateral to ovary up to ventral sucker. Vesicula seminalis elongated 0.33-0.54 mm long, 0.05-0.06 mm wide. Pars-prostatica long, narrow, tubular, 0.54-0.98 mm long, surrounded by large number of prostate gland cell. Ovary spherical, sub-median, pre-testicular, inter-caecal, little anterior to anterior testis, 0.10-0.16 mm long, 0.09-0.17 mm wide at 0.97-1.76 mm from anterior extremity. Vitella- 496 Available at: http://ijpa.tums.ac.ir

Mishr et al.: Two New Trematodes of Family Acanthocolpidae Luhe ria follicular along side of body, extending from middle of cirrus sac up to hind end of body, Uterus pre-ovarian, Metaterm muscular, short, Eggs elongated, elliptical, non-operculated, 0.03-0.04 mm long, 0.01-0.01 mm wide. The present form closely resembles with A. tenuis (3), A. cabelleroi (5), A. inglisi (12), A. equualai, A. lutijanusi (8), A. arpitai, A. apoorvai (13) in having well developed acetabular peduncle. The present form differ from all they above form except A. inguisi and A. cabelleroi, A. arpitai, A. apoorvai in having body elongated slender, oral sucker terminal, sub-spherical, uterine coil not present in between ovary and testis. However present form differs from A. inguisi, A. cabelleroi, A. arpitai and A. apoorvai in having genital pore lying posterior to ventral instead of at the level of pharynx, cirrus sac extending up to ventral sucker instead of level of pharynx, testes are separated from each other by vitellaria instead of lying one behind other in A. inguisi, genital pore lying anterioventral to ventral sucker in A. cabelleroi, genital pore present at level of pharynx ovary preequatorial instead of post equatorial, cirrus sac long instead of short, egg non-operculated instead of operculated in A. arpitai the genital pore lying on posterior to intestinal bifurcation, cirrus sac extending from just in front of ovary instead of at level of vitellaria in A. apoorvai, and in relative shape and size of various organs. Thus an account of above mentioned differences as against all those described earlier, the present form may deserve the status of new species with specific name A. amarawatai sp.nov. Acknowledgements Fig.2: Acanthocolpus amarawatai sp. nov.; 2.1 Ventral view of adult; 2.2 Eggs Discussion Authors are thankful to Head, Department of Zoology, University of Lucknow, Lucknow for providing laboratory facility and Prof K.C. Pandey, Ex-Vice Chancellor, Meerut University and Ex-Head, Department of Zoology, University of Lucknow for giving important suggestions during the work. The authors declare that there is no conflict of interest. References 1. Luhe M. Trematode parasites from marine fishes of Ceylon. Ceyl Pearl Oyst Fish Rep. 1906; 5: 97-108. 2. Srivastava HD. Three new parasites of the genus Acanthocolpus Luhe, 1906. Indian J Vet Sci Amim Husb. 1939; 9:213-216. 3. Manter HW. Studies on digenetic trematodes of fishes of Fiji. III. Family: Acanthocolpidae, Available at: http://ijpa.tums.ac.ir 497

Fellodistomidae and Cryptogonomidae. J Parasit. 1963; 49: 443-450. 4. Fernandes, BM, Zouzu. On a new species of Acanthocolpus. Memo De Inst Oswald. 1973; 241 pp. 5. Gupta AN, Sharma PN. Studies on digenetic trematodes from marine fish of indian waters. III. Ann. Instt. De Biologia Maxico. 1972; 43:93-102. 6. Gupta V, Puri M. On three new species of the genus Acanthocolpus Luhe, 1906 (Trematode: Acanthocolpidae) from marine fishes of Puri, Orissa. Indian J Helminth. 1980; 32:129-137. 7. Verma PS, Singh J. Two new species of genus Acanthocolpus Luhe, 1906 (Digenia: Acanthocolpidae) from marine fishes of Trivendrum, Kerala. Kanpur University Research Journal (Science). 1987; 4:37-40. 8. Gupta SP, Srivastava AB. Studies on digenetic trematode parasites from marine fishes of Rameshwaram, Tamil Nadu. Fotedar commemoration volume. 1989; 93-101. 9. Lokhande LU. Acanthocolpus microtesticulus n.sp. from intestine of the Chirocentrrus dorab in Indian. J Vet Parait. 1990; (4): 55 58. 10. Yamaguti S. Systema Helminthum. Part-I. Digenetic trematodes of fishes. Published by Author. Tokyo 1954: 405. 11. Hafeezullah M. On some new and known digenetic trematodes from marine fishes of India. Indian Journal Helminth. 1971; 45: 73 88. 12. Gupta PC, Gupta SP. On a new trematode Parallelestis bubulcusi n.sp. (family: Echinostomidae Poche, 1920 from the liver of cattle egret, Bulbulcus ibis(linn.) from Udaipur, Rajasthan. Indian J Helminth. 1980; 32: 102-106. 13. Saxena AM, Prakash S. A new species of the genus Lecithochirium Luhe, 1901 from marine fishes, Deegha (W.B.). National J life Science. 2003; 3(1): 97-99. 498 Available at: http://ijpa.tums.ac.ir