On Some Metacercariae and Adult Trematodes of Fishes

Doctorate Research Award-2014 in Parasitology

ON SOME METACERCARIAE AND ADULT TREMATODES OF FISHES

Barrister Kumar Gupta Department Of Zoology University Of Lucknow, Lucknow

Supervisor

Prof. Nirupama Agarwal

International Library for Thesis

Indexed on: December, 2014

UBN : 015-A94510112008 1

ON SOME METACERCARIAE AND ADULT TREMATODES OF FISHES

THESIS SUBMITTED FOR THE AWARD OF DEGREE OF DOCTOR OF PHILOSOPHY IN ZOOLOGY

AT THE UNIVERSITY OF LUCKNOW, LUCKNOW

BY BARRISTER KUMAR GUPTA

M. Sc.

DEPARTMENT OF ZOOLOGY UNIVERSITY OF LUCKNOW, LUCKNOW JUNE, 2011

CONTENTS

Pages

Acknowledgements Introduction 8-9 Material and methods 10 Historical review 11-13

Part I: Metacercaria

1. Neascus bhopalensis n. sp. 15-18 2. Neascus dohrighatensis n. sp. 19-21 3. Neascus khurramnagarensis n. sp. 22-24 4. Neascus kaisarbaghensis n.sp. 25-27 5. Tetracotyle bhopalensis n. sp. 28-30 6. Tetracotyle mauensis n. sp. 31-33 7. Tetracotyle allahabadensis n. sp. 34-36 8. Tetracotyle madhubanensis n. sp. 37-39 9. Tetracotyle saiensis n. sp. 40-42 10. Tetracotyle daliganjensis n. sp. 43-45 11. Tetracotyle megapseudosuckerai n. sp. 46-48 12. Tetracotyle multilobulata n. sp. 49-51 13. Tetracotyle varanasiensis n. sp. 52-54 14. Tetracotyle trilobulata n. sp 55-57 15. Metacercaria of Bucephalopsis garuai Verma, 1936 58-60 16. Metacercaria of B. linguiformis Chakrabarti and Baugh, 1974 61-63 17. Metacercaria of Orchipedum Braun, 1901 64-66 18. Metacercaria of Opisthorchis elongatus Agrawal, 1975 67-69 19. Plagiorchiid metacercaria 70-72 20. Metacercaria of Ommatobrephus Mehra, 1928 73-75 5

21. Metacercaria of Isoparachis hypselobagri (Billet, 1898) Odhner, 1927 76-78 22. Metacercaria of Genarchopsis goppo Ozaki, 1925 79-81

Part II: Adult Trematodes

1. Bucephalus bhagalpurensis n. sp. 83-85 2. Bucephalopsis karvei Bhaleroa, 1937 86-88 3. Bucephalopsis fusiformis Verma, 1936 89-91 4. Allocreadium itwarensis n. sp. 92-94 5. Allocreadium catlai Kakaji, 1969 95-97 6. Allocreadium badatalabensis n. sp. 98-100 7. Eucreadium madhubanensis n. sp. 101-103 8. Rhynchocreadium dohrighatensis n. sp. 104-106 9. Neopodocotyle dohrighatensis n. sp. 107-109 10. Peracreadium bhopalensis n. sp. 110-112 11. Genarchopsis goppo Ozaki, 1925 113-115 12. Haplorchoides varanasiensis n. sp. 116-118 13. Opisthorchis pedicellata Verma, 1927 119-121 14. Oudhia itwaraensis n. sp. 122-124 15. Phyllodistomum vachius Dayal, 1949 125-128 16. Pleuragenoid madhubanensis n. sp. 129-131

References 132-169 Summary 170-175

ACKNOWLEDGEMENT

One of the special advantages I have had is my access to the extraordinary community of researchers at the University. I express my profound gratitude to my Teacher and my Supervisor Prof. Nirupama Agrawal, for her valuable suggestions, consistent advice and scholary critics throughout my research tenure, who corrected my misconceptions and in many cases reviewed chapters. It is a great privilege to express my deep gratitude and indebtedness to Prof. K. C. Pandey, (My PI, DST sponsored project) Department of Zoology, University of Lucknow for his encouragement, and help during the study has kept me on track if not always on schedule, showing inordinate patience. The new art for this thesis includes work from Fine Line Illustrations, coordinated by him. I have been gratified to witness the process by which my often indecipherable scribbles have been transformed into the elegant figures that grace this thesis. My sincere thanks are due to Dr. P. K. Tyagi Reader, Vardhman College, Department of Zoology, Bijnor for advice, moral support and help. It was a great privilege to work with so talented and energetic research colleagues. My colleagues provided me with timely data, critiqued sections of the text, helped me develop figures, answered my questions, and helped in innumerable ways. Space precludes a detailing of every contribution, but I am especially grateful to the following colleagues Dr. Sasikant Shukla, Dr. Richa shukla, Dr. Amita Devak, Dr. Anuradha Pandey, Dr. R. K. Sharma, Dr. Kaushal kishor Singh, Mr. P. G. Yadav, Ms. Ritika Srivastava, Mr. Shailendra Ray, Ms. Amrin Ali, Ms. Saroj and Mr. Lavkush Kumar. The thesis has benefited enormously from the combined wisdom and experience of countless reviewers. I, of course, assume full responsibility for errors of fact or emphasis. I am indebted to all of them for their creative input. Constructive challenges inspired many of the improvements in this thesis. 7

I should not forget the selfless love of my grand mother and bereaved soul of my grand father whose virtual presence inspired me to proceed with my task and other members of the family who endured my odd time working besides providing me financial support. Thanks to all my family members for their support to build up my academic carrier through constant inspiration. My heartiest thanks to my father Shri Ram Gupta, mother Kama Devi for their constant encouragement and blessings. I express my respect and honour to my Father-in-law Mr. Om Prakash, Mother-in-law Mrs. Mithilesh Devi and brother-in-law Pankaj Gupta and Amit Jaiswal for their inspiration and encouragement during research work, so that I could provide much time for preparation of the manuscript and complete the thesis. Also I have no words to express my feelings for my elder brother Mr. Balmiki Gupta, his wife Mrs. Savita Gupta for their enthusiastic inspiration and help alongwith their daughter Barsha for her pleasant presence. My writing has been supported by many people whose advice and encouragement were critical throughout the years of research, my uncles and aunties are some of them. The final manuscript benefited from the superb copyediting by my better-half Priyanka Gupta and my younger brother Dhananjay Gupta. They have all had much to do with the quality of the thesis before you. I shall be failing in my duty if I do not show gratefulness to my friend circle Nirpendar, Sonu, Ashish, Gaurav, Anjali, Nandita, and Bijayata. This is an opportune time to express my thankfulness to all the members of the Deptartment of Zoology, Lucknow University who cooperated in many ways. Financial assistance for this study has been provided by DST, New Delhi also gratefully acknowledged. My sincere apologies to all the wonderful people I have missed. Barrister Kumar Gupta 8

INTRODUCTION

Helminth studies in India has developed enormously during last forty years or so due to contributions of earlier worker like late Mehra, Thapar, Bhalerao, Srivastava, Lal, Dayal, Johari, Pande, Chatterjee, Gogate, Gupta, Singh, and some of the recent worker like Hanumantha Rao, Pandey, Agrawal, Singh, Srivastava, Sood, Ghosh, Tandon, Madhavi and many others. However in comparison to other helminth groups like cestodes, nematodes and acanthocephalan, trematodes have received the maximum attention. Morphology and systematics have been the two main fields of investigation in the study of trematodes by above workers.

Adult trematodes are found infecting almost all vertebrate groups ranging from fishes, amphibians, reptiles, birds and mammals. Their larval form metacercariae are also found infecting fishes, amphibians, reptiles, and some times even birds.

Fish provides nutritious food and are important source of fresh animal proteins. However, almost all of them carry infections of adult trematodes or metacercariae. They infect all the body parts and cause diseases thus reducing their food value. In case of heavy infections, mortality is also caused, which in turn is a great loss to fish industry. They can also transfer infections to man when infected with trematodes larvae i.e. metacercariae. They are dangerous parasites, primarily of carps and siluroids, causing massive epizootics.

In view of growing importance of fish as food in our country, it was suggested by Prof. Nirupama Agrawal to investigate trematode infection of fishes. Since a colleague of mine was already working on trematodes of marine fishes, I selected the freshwater fish for my study. During course of study from 2007-2010 fishes were therefore, examined and author has 9

succeeded in making a collection of some metacercariae and adult trematode parasites which form the basis of the present work.

The thesis has been divided in to two parts. Part I deals with description of metacercariae while Part II deals with adult trematodes. Some of them are new to science while others are recorded only. In the beginning of the thesis, material and methods, applied for the work and a brief history of Indian work done have been given. In the end, a list of references consulted for the work is also appended.

MATERIAL AND METHODS

Fish for the present work were collected from the water bodies of districts Aazamgarh, Allahabad, Ballia, Basti, Belthara, Bhagalpur, Bhatpar Rani, Bhopal (M. P.), Bijnor, Chapra (Bihar), Deoria, Dohrighath, Faizabad, Gonda, Gorakhpur, Jhansi, Kanpur, Lakhimpur Khieri, Lucknow, Madhuban, Mau, Merrut, Pallia, Sahi (U. P.) and Varanasi, and fish markets during 2007- 2011. They were identified using fish base (Froese R., Pauly D. 2007). Their visceral organs were slit open and washed thoroughly in petridishes, containing normal saline. Other organs like body muscles, scales, fins, eye, nasal organ, gills, gall bladder, liver, air bladder, and urinary bladder were also examined thoroughly under dissecting microscope. The worms collected were washed in distilled water and transferred to clean petridishes containing normal saline. They were then studied alive. Subsequently, they were fixed in 70% alcohol under slight pressure of cover slip with grease applied at corners for 24 hours. In case of prolonged storage, 70%, alcohol was used. After fixation, they were washed in distilled water, stained with aquous Aceto-alum Carmine, differentiated in acid water, dehydrated in ascending grades of alcohol, cleared in Clove oil and mounted in Canada Balsam. Drawings were made using drawing tube, attached to a phase contrast microscope (Image analyzer Model BX-51). The measurements in millimeter were taken with the help of an oculometer.

A BRIEF HISTORY OF INDIAN WORK DONE ON ADULT TREMATODES AND METACERCARIAE OF FISHES

Southwell (1913), reported the metacercaria viz; “Distomum” species from Nandus marmoratus (Ham.); and Isoparorchis trisimilitubis from Wallago attu (Bl. and Sch.) and Barbus tor (Ham.) at Calcutta. Later, Southwell and Prasad (1918) described Clinostomum piscidium from Nandus nandus (Ham.) and Trichogaster fasciatus (Bl. and Sch.) at Khulna. Faust (1927) described some strigeid larvae from fishes, after about a decade, viz. Diplostomum schizothoracis from Shizothorax zarudnyi (Nikols.); Schizothorax curvifrqns (Heck.) and Crossocheilus latia (Ham.) respectively, in Kashmir. Subsequently Bhalerao (1926, 1932, 1942a, 1942b, 1943); Srivastava (1950); Chauhan (1947); Kaw (1950a, 1950b); Abraham and Anantaraman (1955); Agarwal (1955, 1958a, 1958b, 1959); Singh (1956, 1959); Premavati (1956); Singh (1956a, 1956b, 1956c); Jaiswal (1957); Khera (1958); Bhardwaj (1961); Rai (1971a, 1971b, 1971c, 1971d); Ganapati et. al. (1962); Gupta, (1963); Pande and Rai (1964); Trivedi (1964); Sinha (1964); Rai and Pande (1965a, 1965b, 1969a, 1969b); Pande et al. (1964); Matta and Pande (1966); Pandey (1968a, 1968b, 1969a,1969b, 1970a, 1970b, 1970c, 1970d, 1971a, 1971b, 1971c, 1971d, 1973a, 1973b, 1973c, 1973d); Thapar (1967); Chakrabarti, (1968a, 1968b, 1970a, 1970b, 1970c, 1974,); Pandey and Baugh (1969, 1970); Rai and Pande (1969a, 1969b); Nath and Pande (1970); Chakrabarti and Baugh (1970, 1973, 1974); Matta and Rai (1971); Pande and Shukla (1972a, 1972b, 1973, 1974); Dutta and Srivastava (1972); Madhavi (1978a, 1978b); Agarwal (1975a, 1975b); Pandey and Agarwal (1977); Agrawal and Singh (1980); Dwivedi and Dwivedi (1981); Agarwal and Khan (1982a, 1982b, 1982c); Tewari (1982); Chopra, Kumar and Singh (1983); Pandey and Tewari (1983, 1984, 1986); Pandey and Tiwari(1983); Pandey and Tyagi (1986); Tewari and Tyagi (1986); Shameem and Madhavi (1988); Sinha, Sinha and Nikhil (1988); Madhavi and Shamim (1993); Thakur and Prasad (1997); Pandey and Pandey (2000); Pandey and Kiran, 2002; Fatima 12

and Gupta (2008), Pardeshi & Hiware (2010); and have worked on trematode larvae, the metacercariae of fishes in India. Credit goes to Verma (1927) who described the first adult piscine trematoda Opisthorchis pedicellata from Bagarius yarrellii (Syk.) and Rita rita (Ham.) and a number of Bucephalid worms from Allahabad. Thapar (1930) added a new genus Gomtia from Bagarius yarrellii (Syk.) at Lucknow. Subsiquent workers who contributed on taxonomy of adult digenetic trematodes of fishes in India are Mehra (1941); Bhalerao, (1936a,1936b, 1937, 1941, 1942a, 1942b, 1943); Verma, (1927a, 1927b, 1936a, 1936b); Dayal (1930, 1935, 1938a, 1938b, 1938c, 1938d, 1938e, 1939, 1942, 1944, 1948a, 1950, 1952); Harshey (1933, 1937); Chatterji (1933); Srivastava (1933, 1934, 1935a, 1935b, 1935c, 1936, 1938a, 1938b, 1938c, 1939); Pande (1934, 1937a, 1937b, 1938a, 1938b); Thapar (1930, 1960); Chauhan (1943, 1947, 1949, 1953a, 1953b,); Kaw (1943, 1944,); Chatterji (1953a, 1953b, 1957); Singh, (1956a, 1956b); Gupta, (1950, 1951a, 1951b, 1951c, 1951d, 1951e, 1953a, 1953b, 1955,1956a, 1956b, 1958, 1961, 1963); Dayal and Gupta (1953); Baugh (1956, 1958, 1963); Jaiswal (1957); Agarwal (1959); Gupta and Srivastava (1960); Srivastava (1960); Motwani and Srivastava (1961a); Rai (1962, 1964a, 1964b); Srivastava (1962a, 1962b, 1963, 1968, 1982); Agarwal (1963, 1964a, 1964b, 1966); Simha and Prasad (1964); Prasad (1965); Gupta and Agarwal (1966, 1967); Srivastava and Singh (1967); Srivastava and Ghosh (1967); Rai and Pande (1968); Kakaji, (1968, 1969a, 1969b); Sircar and Sinha (1969); Chauhan and Dwevedi (1969); Rai (1971a, 1971b, 1971c, 1971d); Pandey (1969, 1970a, 1970b, 1971, 1973a, 1973b, 1975); Baugh and Chakrabarti (1970); Gupta and Kumari (1970a, 1970b, 1970c, 1970d); Agarwal and Verma (1972, 1981); Verma (1973a, 1973b) Singh and Sinha (1975, 1976); Vasantha Kumari and Srivastava (1976); Pande and Shukla (1976); Agarwal and Kumar (1977, 1979, 1981, 1983, 1985, 1987); Bhadauria and Dandotia (1977,1979); Ahmad (1985); Agarwal and singh (1978); Madhavi (1978a, 1978b); Agarwal and Singh (1980, 1981); Dandotia and Bhadauria (1979); Singh and Prasad (1978); Agarwal and 13

Agarwal (1980, 1984); Kumar and Agarwal (1980); Ubdage and Agarwal (1980); Pandey and Dewedi (1983); Agarwal and Agarwal (1984); Gupta and Singh (1985a, 1985b); Dhar and Kharoo (1986); Swarup and Jain (1987); Duggal and Bedi (1987); Prakash (1988); Agarwal and Agarwal (1984, 1989); Maurya, Agarwal and Singh (1989a, 1989b, 1993); Pandey and Tiwari (1989); Gupta and Gupta (1990); Agarwal and Sharma (1989, 1991); Maurya and Agarwal (1992,1998a,1998b,1999,2000,2004); Jhansilakshmibai and Madhavi (1997); Umadevi and Madhavi (2000, 2006); Vasandakumar and Janardanan (2002a, 2002b); Govind, Sabhyata, and Gundevia (2004); Sabhyata and Govind (2004, 2008a, 2008b); Pandey, Pandey and Govind (2004); Sen and Siddiqui (2006); Pandey and Govind (2007); Sabhyata and Gundevia (2007a, 2007b); Pardeshi and Hiware (2010).

Part-I Metacercaria

Neascus bhopalensis n. sp. (Figs. 1 - 3)

Host: Xenentodon cancila (Ham.) No. of host examined: 18 No. of host found infected: 03 No. of metacercariae collected: 28 Location: Liver Locality: Itwara fish market, Bhopal

Cyst (Fig.1) oval, two layered, transparent, 0.82 - 0.93 mm x 0.29 - 0.38 mm. Body (Fig.2) aspinose, foliaceous, differentiated into attenuated fore body, 0.63 - 0.88 mm x 0.27 - 0.30 mm, and conical hind body 0.22 - 0.27 mm x 0.18 - 0.20 mm. Oral sucker terminal, oval, 0.03 - 0.07 mm x 0.03 - 0.04 mm. Ventral sucker slightly larger, spherical, located in middle of body, 0.04 - 0.05 mm x 0.05 - 0.06 mm. Hold fast organ strongly developed, oval, situated behind ventral sucker, 0.12 - 0.15 mm x 0.13 - 0.16 mm. Hold fast gland bilobed and surrounds posterior border of holdfast organ. Rudimentary gonads, represented by 3 dark stained cell masses, in hind body; two large transversely elongated masses appear to be presumptive testes of 0.02 - 0.07 mm x 0.06 - 0.12 mm and 0.02 - 0.08 mm x 0.10 - 0.16 mm and one small mass, presumptive ovary in between two testes, 0.01 - 0.03 mm x 0.05 - 0.09 mm. Bursa copulatrix prominent, represented by dark stained cell mass, at hind end of body, 0.06 - 0.12 mm x 0.07 - 0.11 mm. Excretory bladder (Fig.3) ‘V’ shaped and opens at hind end of body. Two main reserve excretory canals arise, one on each side of excretory bladder, run antero-laterally to join an anterior transverse canal behind oral sucker. A median excretory canal arises from anterior transverse canal, descends posteriorly, upto hind end of fore body and joins main canals by a posterior transverse canal. Two more side canals, one on each side of median excretory canal, descend from anterior 16

transverse canal and run posteriorly to join transverse canal. Reserve canals are gorged with free floating oval to round excretory corpuscles.

Discussion

The Indian species of strigeid metacercariae of group Neascus Hughes, 1927 are N. vetastai Kaw, 1950, N. chelai Khera, 1958, N. indicus Thapar, 1967, N. cirrhinus Thapar, 1967, N. elongates (Singh, 1956) Pandey, 1970, N. hepatica Chakrabarti, 1970, N. channi Pandey, 1971, N. xenentodoni Pandey, 1971, N. komiyai Pandey, 1973, N. hoffmani Pandey, 1973, N. gussevi Chakrabarti, 1974, N. nanaksagrensis Baugh and Chakrabarti, 1977, N. chauhani Agrawal and Khan, 1982, N. hanumanthai Agrawal and Khan, 1982, N. simhai Agrawal and Khan, 1982, N. moghei Agrawal and Khan, 1982, N. shahjahanpurensis Pandey and Tiwari, 1986, N. ramalingami Pandey and Tiwari, 1986 and N. vedi Pandey and Tiwari, 1986. The present larva differs from N. vetastai in absence of pharynx and shape of holdfast gland, from N. chelai in ratio of fore and hind body, and absence of pharynx, N. indicus and N. cirrhinus in absence of pharynx and ratio of suckers, from N. elongates in absence of pharynx and number of genital rudiments, from N. hepatica, N. Xenentodoni and N. gussevi in absence of gut, from N. channi in ratio of fore and hind body and shape of holdfast gland, from N. komiyai in ratio of fore and hind body, absence of gut and shape of holdfast gland, from N. hoffmani in arrangement of genital rudiment, from N. nanaksagrensis in absence of pharynx and shape of genital rudiment, from N. chauhani by ratio of fore and hind body and location of genital rudiments, from N. hanumanthai in absence of pharynx and ratio of suckers, from N. simhai by absence of pharynx and shape of holdfast gland, from N. moghei and N. vedi in absence of gut and pharynx, from N. shahjahanpurensis in ratio of fore and hind body and location of holdfast organ, and from N. ramalingami in ratio of suckers and absence of pharynx and gut. 17

Among species known from foreign land, it also differs from Neascus of Uvulifer ambloplitis Hughes, 1927 in ratio of fore and hind body and ratio of sucker, from Neascus of Ornithodiplostomum ptychocheilus Hughes and Piszezek, 1928 in absence of pharynx and gut and ratio of suckers, from Neascus of Posthodiplostomum minimum Hughes, 1928 in absence of gut and pharynx, from Neascus of Crassiphiala bulboglossa Hughes, 1928 in absence of pharynx and gut and ratio of fore and hind body, from Neascus of Neodiplostomum perlatum Ciurea, 1930 in ratio of suckers and absence of pharynx, from Neascus of Posthodiplostomum cuticola Ciurea, 1930 in absence of gut and pharynx, from Neascus grandis Mueller and Van Cleave, 1932 by absence of gut, pharynx and holdfast gland, from Neascus rhinichthysi Hunter and Wanda, 1933 in absence of pharynx, gut and position of genital rudiments, from Neascus ellipticus Chandler, 1951 in absence of pharynx and gut, from Neascus longicollis Chandler, 1951 in ratio of suckers and fore and hind body, from Neascus pyriformis Chandler, 1951 in absence of pharynx and from Neascus nolfi Hoffman, 1955 in ratio of suckers and absence of gut, Therefore, the larva is regarded a new species and named Neascus bhopalensis n. sp. after the locality wherefrom the host was procured.

0.2mm

Fig.1

0.2mm 0.2mm

Fig.2 Fig.3 Neascus bhopalensis n. sp.

Fig.1 encysted metacercaria Fig.2 metacercaria Fig.3 metacercaria, showing reserve excretory system (drawn from live specimens) 19

Neascus dohrighatensis n. sp. (Figs. 4 - 6)

Host: Wallago attu (Bl. & Schn.) Location: Liver Locality: Dohrighat (U.P.) No. of host examined: 05 No. of host found infected: 01 No. of metacercariae collected: 09

Cyst (Fig.4) oval, delicate, thin walled, 0.60 - 0.70 mm x 0.10 - 0.16 mm. Body (Fig.5) elongated, aspinose, 0.54 - 0.60 mm x 0.16 - 0.20 mm; divided into a fore and a hind body. Fore body tongue-like, 0.35 - 0.40 mm x 0.15 - 0.20 mm., hind body oval, devoid of spines, 0.19 - 0.25 mm x 0.10 - 0.14 mm. Oral sucker terminal, elongated, 0.05 x 0.04 mm. Ventral sucker oval, pre-eqatorial, slightly smaller than oral sucker, 0.03 x 0.04 mm. Pre- pharynx, pharynx and intestinal caeca not observed. Hold fast organ strongly developed, 0.07 - 0.08 mm x 0.08 - 0.09 mm, in posterior region of fore body. Hold fast gland bilobed, transversely elongated, 0.06 - 0.07 mm x 0.03 - 0.04 mm. Gonads in form of three densely stained masses. One anterior oval mass, 0.09 - 0.10 x 0.02 - 0.03 mm, appears to be ovary. Two posterior 'V’ shaped, transversely placed masses are presumptive testes, 0.08 - 0.09 mm x 0.02 - 0.03 mm. Bursa copulatrix well developed, 0.05 - 0.06 mm x 0.03 - 0.04 mm, at posterior end of body. Excretory bladder (Fig.6) ‘V’ shaped. Reserve excretory system of two main reserve excretory canals, running one on each lateral side of body upto oral sucker and opening in an anterior transverse canal. A median longitudinal canal runs in median field of body upto holdfast organ and opens into a posterior transverse canal which also joins two main reserve excretory canals. These canals are filled with excretory corpuscles.

Discussion

Of all the known metacercariae of larval genus Neascus Hughes, 1927, the present larva comes closer to N. vitastai, N. indicus, N. cirrihinus, N. hepatica, N. channi, N. xenentodoni, N. komiyai, N. hoffmani, N. gussevi, N. nanaksagrensis, N. chauhani, N. hanumanthai, N. simhai, N. moghei, N. shahjahanpurensis, N. ramalingami and N. vedi in the number of genital rudiments but chiefly differs from them in shape of holdfast organ and holdfast gland. It further differs from N. vitastai in absence of prepharynx, from N. indicus, N. cirrihinus, N. chauhani and N. shahjahanpurensis in position of holdfast organ and holdfast gland and absence of pharynx and gut, from N. hepatica, N. channi, N. xenentodoni, N. komiyai, N. hoffmani, N. gussevi, N. nanaksagrensis, N. simhai, N. moghei, and N. vedi in ratio of suckers and from N. ramalingami in the presence of bursa copulatrix. Among the species described from foreign land, it comes closer to Neascus of Ornithodiplostomum ptychocheilus, Neascus of Neodiplosyomum perlatum, Neascus of Posthodiplostomum cuticola, Neascus of Posthodiplostomum minimum, and Neascus rhinichthysi in number of genital rudiments but differs in shape of body and absence of pharynx and gut. It further differs from Neascus of Ornithodiplostomum ptychocheilus, Neascus of Posthodiplostomum cuticola, Neascus of Posthodiplostomum minimum, in ratio of sucker, ratio of body, from Neascus of Neodiplosyomum perlatum and Neascus rhinichthysi in shape of holdfast organ and holdfast gland. Therefore, the larva is regarded a new species and named Neascus dohrighatensis n. sp. after the locality where from the host was procured.

0.1mm

Fig.4

0.1mm 0.1mm

Fig.5 Fig.6 Neascus dohrighatensis n. sp.

Fig.4 encysted metacercaria Fig.5 metacercaria Fig.6 metacercaria, showing reserve excretory system (drawn from live specimens) 22

Neascus khurramnagarensis n. sp. (Figs.7 - 9)

Host: Xenentodon cancila (Ham.) Location: Liver Locality: Khurram Nagar fish market, Lucknow No. of host examined: 25 No. of host found infected: 15 No. of metacercariae collected: 79

Cyst (Fig.7) oval, thick, transparent, 1.13 - 1.33 mm x 0.90 - 1.20 mm. Body (Fig.8) aspinose, elongated, differentiated into fore and hind body, fore body 0.47 - 1.03 mm x 0.19 - 0.21 mm, body tube like, 0.19 - 0.22 mm x 0.13 - 0.19 mm. Oral sucker terminal, oval, 0.03 - 0.04 mm x 0.02 - 0.03 mm. Ventral sucker elongate-oval, equal to oral sucker, post-eqatorial, 0.03 - 0.04 mm x 0.02 - 0.03 mm. Hold fast organ strongly developed, situated behind ventral sucker, at hind region of fore body, 0.12 - 0.17 mm x 0.10 - 0.16 mm. Hold fast gland bilobed. Pharynx and gut absent. Gonads represented by two dark stained cell masses; one large cresent, 0.02 - 0.07 mm x 0.06 - 0.12 mm, and the other mass, 0.01 - 0.03 mm x 0.05 - 0.09 mm. Bursa copulatrix oval and located in hind body. Excretory bladder (Fig.9) ‘V’ shaped and opens out by terminal excretory pore. Two main reserve excretory canals arise, one on each side of excretory bladder, run laterally, upto oral sucker and join anterior transverse canal. A median excretory canal arises from anterior transverse canal and runs posteriorly to join a posterior transverse canal, in region of ventral sucker. All canals contain excretory corpuscles of different sizes.

Discussion

The present larva closely resembles with N. vetastai, N. hepatica, N. xenentodoni, N. hoffmani, N. hanumanthai, N. simhai, N. moghei, N. 23

ramalingami and N. vedi in shape of body but differs from them in number of genital rudiments. It further differs from N. hepatica and N. vedi in ratio of suckers, from N. vetastai, N. xenentodoni, N. hoffmani, N. hanumanthai, N. simhai, N. moghei and N. ramalingami in shape of holdfast organ and holdfast gland. Among the species described from foreign land, it comes closer to Neascus of Uvulifer ambloplitis, Neascus of Crassiphiala bulboglossa, Neascus of Posthodiplostomum minimum, Neascus of Posthodiplostomum cuticola, Neascus grandis, Neascus pyriformis and Neascus ellipticus in shape of body but differs in absence of pharynx and gut. It further differs from Neascus of Uvulifer ambloplitis, Neascus of Crassiphiala bulboglossa, Neascus of Posthodiplostomum minimum, Neascus of Posthodiplostomum cuticola, Neascus grandis, Neascus pyriformis and Neascus ellipticus in ratio of fore and hind body, shape and position of holdfast organ and holdfast glands. Therefore, the larva is regarded a new species and named Neascus khurramnagarensis n. sp. after the locality wherefrom the host was procured.

0.2mm

Fig.7

0.2mm 0.2mm

Fig.8 Fig.9 Neascus khurramnagarensis n. sp.

Fig.7 encysted metacercaria Fig.8 metacercaria Fig.9 metacercaria, showing reserve excretory system (drawn from live specimens) 25

Neascus kaisarbaghensis n. sp.

(Figs. 10 - 12)

Host: Anabas testudineus (Bl.) Location: Cranium/Branchial region Locality: Kaisarbagh fish market, Lucknow No. of host examined: 05 No. of host found infected: 02 No. of metacercariae collected: 07

Cyst (Fig.10) oval, transparent, 1.13 - 1.33 mm x 0.90 - 1.20 mm. Body (Fig.11) aspinose, foliaceous, well differentiated into fore and hind body, fore body 0.47 - 1.03 mm x 0.19 – 0.21 mm and hind body 0.19 - 0.22 mm x 0.13 - 0.19 mm. Oral sucker terminal, 0.03 - 0.04 mm x 0.02 - 0.03 mm. Ventral sucker smaller than oral sucker, elongate-oval, 0.03 - 0.04 mm x 0.02 - 0.03 mm. Pharynx absent. Intestinal caeca reaching upto posterior end of hind body. Hold fast organ strongly developed, situated behind ventral sucker, 0.13 - 0.15 x 0.17 - 0.20 mm. Hold fast gland bilobed, transversely elongated, located behind holdfast organ, 0.28 - 0.30 mm x 0.02 - 0.04 mm. Gonads represented by three dark stained cell masses; two large masses appear to be testes, 0.02 - 0.07 mm x 0.06 - 0.12 mm and 0.02 - 0.08 mm x 0.10 - 0.16 mm and one small elongated mass in between two testes, 0.01 - 0.03 mm x 0.05 - 0.09 mm, could be future ovary. Copulatory bursa, oval and located in hind body. Excretory bladder (Fig.12) ‘V’ shaped and opens outside terminally through an excretory pore, at hind body region. Two main collecting canals of reserve excretory system originate, one of each side, from cornua of excretory bladder, run through lateral body margins upto level of pharynx where they unite with each other by an anterior transverse canal. A median reserve excretory canal originates from anterior transverse canal, runs posteriorly upto holdfast organ and joins posterior transverse canal which further opens into 26

two main canals. Two lateral canals, on an each side of median canal also originate from anterior transverse canal, runs upto holdfast organ and form posterior transverse canal. Two median longitudinal canals, originate at junction of fore and hind body. These major canals are joined together by short transverse canals but their arrangement could be observed in detait. Small, black, excretory corpuscles flow freely in canals.

Discussion

The present larva closely resembles with N. vetastai, N. hepatica, N. xenentodoni, N. hoffmani, N. hanumanthai, N. simhai, N. moghei, N. ramalingami and N. vedi in shape of body and number of genital rudiments but differs from them in shape of holdfast organ and holdfast gland. It further differs from N. vetastai, N. xenentodoni, N. hoffmani, N. simhai and N. moghei in ratio of suckers, from N. hepatica and N. vedi in ratio of fore and hind body, from N. hanumanthai and N. ramalingami in absence of pharynx. Among the species described from foreign land, it comes closer to Neascus of Ornithodiplostomum ptychocheilus, Neascus of Posthodiplostomum minimum, Neascus of Posthodiplostomum cuticola, Neascus grandis, Neascus rhinichthysi, Neascus ellipticus, and Neascus pyriformis in shape of body but differs in absence of pharynx and ratio of body. It further differs from Neascus grandis, Neascus pyriformis and Neascus ellipticus in number of genital rudiments, from Neascus of Ornithodiplostomum ptychocheilus, Neascus of Posthodiplostomum minimum and Neascus of Posthodiplostomum cuticola in ratio of suckers and from Neascus rhinichthysi in shape of holdfast organ and holdfast gland. Therefore, the larva is regarded a new species and named Neascus kaisarbaghensis n. sp. after the locality wherefrom the host was procured. 27

0.2mm

Fig.10

0.2mm 0.2mm

Fig.11 Fig.12 Neascus kaisarbaghensis n. sp. Fig.10 encysted metacercariae Fig.11 metacercaria Fig.12 metacercaria, showing reserve excretory system (drawn from live specimens) 28

Tetracotyle bhopalensis n. sp. (Figs. 13 - 15)

Host: Channa punctatus (Bloch) Location: Gut Locality: Fish market, Bhopal No. of host examined: 20 No. of host found infected: 05 No. of metacercaria collected: 10

Cyst (Fig.13) oval, thin, transparent, 1.02 - 1.05 mm x 0.66 - 0.68 mm. Body (Fig.14) elongated, aspinose, 1.25 - 1.27 mm x 0.63 - 0.64 mm. Suckers well developed and muscular. Oral sucker sub-terminal, more or less circular, 0.10 mm - 0.11 mm. Ventral sucker pre-equatorial, larger than oral sucker, 0.11 - 0.12 mm x 0.12 - 0.13 mm. Prepharynx absent. Pharynx elongate-oval, 0.04 - 0.05 mm x 0.04 - 0.06 mm. Pseudo-suckers situated on each side of intestinal bifurcation, 0.15 - 0.17 mm x 0.09 - 0.10 mm. Hold fast organ well developed, 0.36 - 0.38 mm x 0.21 - 0.27 mm, elongate, 3 lobed, with a distinct ‘U’ shaped cavity. A ‘U’ shaped holdfast gland, behind holdfast organ present. Genital rudiment represented by two tandem, elongated cell masses, at hind region of body. Excretory bladder (Fig.15) ‘V’ shaped, located at posterior end of body, opening outside by terminal excretory pore. Main reserve excretory canals, one on each lateral side, run anteriorly upto pseudosuckers and join fellow canal by a transverse canal. A median excretory canal, runns posteriorly, in median region of body, upto gonads and joins main canal. Canals are filled with free floating, round to oval corpuscles.

Discussion

To the best of my knowledge, the following species of metacercariae are known under strigeid group Tetracotyle Faust, 1918 from India viz. T. 29

ranae Kaw, 1950; T. sophorensis Singh, 1956; T. indicus Singh, 1956; T. ujjainensis Trivedi, 1964; T. szidati Chakrabarti and Baugh, 1970; T. xenentodoni Chakrabarti, 1970; T. aglandulata Baugh and Chakrabarty, 1970; T. muscularis Chakrabarty, 1970; T. glossogobii Chakrabarti, 1970; T. lali Pandey, 1970; T. lucknowensis Pandey, 1971; T. singhi Pandey, 1973; T. baughi Pandey, 1973; T. tandani Pandey, 1973; T. bufoi Agrawal, 1975; T. lymnaei Pandey and Agrawal, 1978; T. gyanpurensis, Agrawal and Singh, 1980; T. pandei Agrawal and Khan, 1982; T. srivastavi Agrawal and Khan, 1982; T. ramalingi Agrawal and Khan, 1982; T. simhai Pandey and Tiwari, 1983; T. sanjivi Pandey and Tiwari, 1983; T. fotedari Pandey and Tiwari, 1983; T. satendri Tiwari and Tyagi, 1986; T. kawi Pandey and Tyagi, 1987; T. kalyani Pandey and Pandey, 2000; and T. satyapalii Pandey and Pandey, 2000. Of the above species, the present larva closely resembles with T. ranae, T. indicus, T. ujjainensis, T. lali, T. singhi, T. gyanpurensis, and T. satyapali in position of pseudosuckers and holdfast organ but differs from them in the number of genital rudiments which is two in the present larva. It further differs from T. indicus, T. singhi, and T. satyapalii, T. ujjainensis and T. gyanpurensis in the shape of holdfast organ, and from T. ranae and T. lali in shape of body which is elongate-oval. Among the species described from foreign land, it differs from Tetracotyle diminuta Hughes, 1928 in ratio of sucker and position of pseudosuckers, Tetracotyle of Apatemon fuligulae Yamaguti, 1933 and Tetracotyle of Apatemon pellucidus Yamaguti, 1933 in shape of body, absence of prepharynx, oesophagus and intestinal caeca, from Tetracotyle of Cotyluris communis Hughes, 1928, Tetracotyle biwaensis Yamaguti, 1942 and Tetracotyle tahoensis Haderlie, 1953 in number of genital rudiments, shape of holdfast organ and holdfast glands. Therefore, the larva is regarded new to science and named T. bhopalensis n. sp. after the locality where from the host was obtained.

0.2mm

Fig. 13

0.2mm 0.2mm

Fig.14 Fig.15

Tetracotyle bhopalensis n. sp.

Fig.13 encysted metacercaria Fig.14 metacercaria Fig.15 metacercaria, showing reserve excretory system (drawn from live specimens) 31

Tetracotyle mauensis n. sp.

(Fig. 16 - 18)

Host: Xenentodon cancila (Ham.) Location: Liver/Gut Locality: Mau (U.P.) No. of host examined: 18 No. of host found infected: 02 No. of metacercaria collected: 05

Cyst (Fig.16) oval, thick, 1.22 - 1.24 mm x 0.75 - 0.76 mm., two layered, outer layer thick, fibrous, tough, pigmented and inner layer thin, transparent. Body (Fig.17) undivided, aspinose, with narrow anterior and broad posterior ends, 0.33 - 0.43 mm x 0.24 - 0.29 mm. Oral sucker 0.09 - 0.11 mm x 0.09 - 0.10 mm. Ventral sucker smaller than oral sucker, 0.04 - 0.05 mm x 0.04 - 0.05 mm. Pseudo-suckers large, muscular, somewhat triangular, located at lateral sides of pharynx, 0.09 - 0.15 mm x 0.09 - 0.10 mm. Oesophagus and intestinal caeca not visible. Hold-fast organ well developed, pear shape, posterior to ventral sucker, having a distinct cavity. A dark mass of cells, representing holdfast gland, at hind end of hold fast organ present. Genital rudiment, represented by an elongate-oval, dark stained cell mass at hind region of body, overlap the holdfast gland. Excretory bladder (Fig.18) ‘V’ shaped. Inner and outer longitudinal canals are joined in region of pseudo-suckers. 9 - 11 transverse canaliculae join to outer longitudinal canals. Reserve excretory system filled with round excretory corpuscles.

Discussion

The present larva closely resembles with T. glossogobii, T. lucknowensis, T. kawi, T. sanjivi, T. pandei and T. satendri in shape of body 32

and shape and position of pseudosuckers but differs from them in ratio of suckers. It chiefly differs from T. glossogobii, T. lucknowensis, T. sanjivi, T. pandei and T. satendri in the number of genital rudiments, from T. kawi in shape of holdfast organ and holdfast gland. It also differs from T. bhopalensis n. sp. Described in earlier pages, in shape, size and position of pseudosuckers, and also in the number of genital rudiments.

Among the species described from foreign land, it comes closer to Tetracotyle of Cotyluris communis, T. diminuta, Tetracotyle of Apatemon pellucidus, Tetracotyle biwaensis and Tetracotyle tahoensis by the absence of prepharynx and position of pseudosuckers but differs in shape of holdfast organ and holdfast gland. It further differs from Tetracotyle of Cotyluris communis, T. diminuta, Tetracotyle biwaensis and Tetracotyle tahoensis in absence of esophagus and intestinal bifurcation and ratio of suckers, from Tetracotyle of Apatemon fuligulae in absence of prepharynx and shape of holdfast organ and holdfast gland and from Tetracotyle of Apatemon pellucidus in the presence of pharynx and ratio of body. Therefore, the larva is regarding as new to science and named Tetracotyle mauensis n. sp. after the locality wherefrom the host was procured.

0.1mm

Fig.16

0.1mm 0.1mm

Fig.17 Fig.18

Tetracotyle mauensis n. sp.

Fig.16 encysted metacercaria Fig.17 metacercaria Fig.18 metacercaria, showing reserve excretory system (drawn from live specimens) 34

Tetracotyle allahabadensis n. sp. (Fig. 19 - 21)

Host: Mastacembelus puncalus (Ham.) Location: Gut Locality: Allahabad No. of host examined: 15 No. of host found infected: 03 No. of metcercaria collected: 07

Cyst (Fig.19) elongate oval, 0.84 - 0.90 mm x 0.55 - 0.61 mm. Body (Fig.20) aspinose, ovoidal, 1.23 mm x 0.70 mm, fore body cup-shaped 0.90 mm x 0.70 mm, hind body somewhat cylindrical but short, 0.20 - 0.25 mm x 0.25 - 0.30 mm. Suckers well developed, circular, unequal. Oral sucker terminal, more or less circular, 0.15 - 0.12 mm. Ventral sucker circular, slightly larger than oral sucker, in middle of body, 0.17 - 0.16 mm. Pseudosuckers prominent, muscular, oval, on either side of esophagus, 0.26 - 0.32 mm x 0.16 - 0.19 mm. Prepharynx absent. Pharynx subglobular, feebly muscular, 0.03 - 0.05 mm x 0.02 - 0.04 mm. Oesophagus short. Intestinal bifurcation obscured. Genital rudiments represented by two tandem, elongate cell masses of dark staining cells, at posterior end of body. Anterior mass may be presumptive testis, 0.05 - 0.06 mm x 0.10 - 0.12 mm, while posterior mass ovary, 0.04 - 0.06 mm x 0.11 - 0.12 mm. Hold fast organ well developed, bowl-shaped, lobes protrusible and overlap each other, 0.40 - 0.48 mm x 0.15 - 0.17 mm, enclosing ventral sucker. Hold fast gland in just close to holdfast organ. Excretory bladder (Fig.21) ‘V’ shaped, excretory pore terminal. Main reserve excretory canals, one on each side of body run upto oral sucker and join each other by a transverse canal. A median canal runs upto the holdfast organ and joins two more canals by a posterior transverse canal. Small and round excretory corpuscles of different sizes flow in excretory canals.

Discussion

The present larva closely resembles with T. sophorensis, T. indicus, T. xenentodoni, T. glossogobii, T. singhi and T. srivastavi in shape of body, position and shape of pseudosuckers but differs from them in shape of holdfast organ and holdfast gland. It differs from T. indicus, T. xenentodoni T. singhi and T. srivastavi in the number of genital rudiments and from T. sophorensis and T. glossogobii in position of holdfast organ and holdfast gland. Among the species described from foreign land, it comes closer to Tetracotyle of Cotyluris communis, Tetracotyle of Apatemon fuligulae, Tetracotyle of Apatemon pellucidus, Tetracotyle biwaensis and Tetracotyle tahoensis in ratio of suckers but differs in number of genital rudiments. It further differs from Tetracotyle of Cotyluris communis, Tetracotyle biwaensis and Tetracotyle tahoensis in shape of body and absence of intestinal bifurcation, from Tetracotyle of Apatemon fuligulae in absence of prepharynx, shape of holdfast organ and holdfast gland and from Tetracotyle of Apatemon pellucidus in the presence of pharynx and ratio of body. Therefore, the larva is regarded a new species and named Tetracotyle allahabadensis n. sp. after the locality where from the host was collected.

0.2mm

Fig. 19

0.2mm 0.2mm

Fig.20 Fig.21

Tetracotyle allahabadensis n. sp.

Fig.19 encysted metacercaria Fig.20 metacercaria Fig.21 metacercaria, showing reserve excretory system (drawn from live specimens) 37

Tetracotyle madhubanensis n. sp.

(Fig. 22 - 24)

Host: Mastacembelus puncalus (Ham.) Location: Branchial region Locality: Fish market, Madhuban No. of host examined: 10 No. of host found infected: 03 No. of metacercaria collected: 06

Cyst (Fig.22) oval, thick, transparent, devoid of pigments. Body (Fig.23) aspinose, broadly oval, divided into a fore body, 0.80 - 1.00 mm x 0.85 - 1.03 mm and a short hind body, 0.25 - 0.27 mm x 0.50 - 0.53 mm. Oral sucker terminal, round to oval, 0.10 - 0.13 mm x 0.15 - 0.19 mm. Ventral sucker equatorial, smaller than oral sucker, 0.13 - 0.14 mm 0.13 - 0.14 mm. Lateral Pseudosuckers large, prominent, oval, covering almost entire antero- lateral region of body, extending upto ventral sucker, 0.47 - 0.53 mm x 0.22 - 0.26 mm. Prepharynx present. Pharynx leading into oesophagus, 0.08 - 0.09 mm x 0.09 - 0.10 mm. Intestinal caeca extending up to holdfast organ. Hold fast organ located close to ventral sucker, bilobed, protrusible. Hold fast gland present. Genital rudiments represented by three transvesrsally elongated post acetabular masses, located one behind other. Anterior and posterior masses may be presumptive testes while median mass ovary. Anterior mass 0.25 - 0.30 mm x 0.03 - 0.05 mm, median mass 0.18 - 0.20 mm x 0.02 - 0.04 mm, and the posterior mass 0.10 - 0.13 mm x 0.02 – 0.04 mm. Excretory bladder (Fig.24) ‘V’ shaped, excretory pore terminal. Each cornua of excretory bladder gives rise to a main reserve excretory canal, on each lateral side of body, reaching up to region of pharynx and joined together by an anterior transverse canals. From anterior transverse canal descend two lateral longitudinal canals, one on each medial side of main collecting canal, opening into posterior transverse excretory canal which also join the main collecting 38

canals at margin of fore and hind body. Each lateral longitudinal canal further join main collecting canal of its side by 08-10 short, transverse ducts. Furthermore, the main canals extend anteriorly on each lateral side upto oral sucker in the form of large broad ducts. Whole reserve system filled with freely moving, small, rounded excretory corpuscles.

Discussion

The present larva closely resembles with T. sophoriensis, T. ujjainensis, T. muscularis, T. glossogobii, T. lucknowensis, T. baughi, T. tandani, T. chauhani and T. kalyani in the number of genital rudiments, position of pseudosuckers but differs from them in the shape of holdfast organ and holdfast gland. It further differs from T. sophoriensis, T. chauhani and T. kalyani in the shape of body, from T. ujjainensis, T. glossogobii, T. lucknowensis, and T. baughi in the position of holdfast organ and holdfast gland and from T. muscularis and T. tandani in ratio of fore and hind body and suckers. Among the species described from foreign land, it comes closer to Tetracotyle diminuta, Tetracotyle of Cotyluris communis, Tetracotyle of Apatemon fuligulae, Tetracotyle of Apatemon pellucidus, Tetracotyle biwaensis and Tetracotyle tahoensis in shape of body and presence of esophagus and intestinal caeca but differs in shape of holdfast organ and holdfast gland. It further differs from Tetracotyle of Cotyluris communis, Tetracotyle of Apatemon fuligulae, Tetracotyle biwaensis and Tetracotyle tahoensis in number of genital rudiments, from Tetracotyle of Apatemon pellucidus and Tetracotyle diminuta in ratio of suckers. Therefore, the larva is regarded a new species and named Tetracotyle madhubanensis n. sp. after the locality where from the host was procured.

0.3mm

Fig.22

0.2mm 0.2mm

Fig.23 Fig.24

Tetracotyle madubanensis n. sp.

Fig.22 encysted metacercaria Fig.23 metacercaria Fig.24 metacercaria, showing reserve excretory system (drawn from live specimens) 40

Tetracotyle saiensis n. sp. (Fig. 25 - 26)

Host: Colisa fasciatus (Bl. & Schn.) Location: Body cavity Locality: River Sai, Lucknow No. of host examined: 20 No. of host found infected: 01 No. of metacercaria collected: 03

Cyst (Fig.25) oval, single layered, non pigmented, transparent, 0.81 - 0.85 mm x 0.52 - 0.68 mm. Body (Fig.26) aspinose, oval, 1.0 - 1.12 mm x 0.49 - 0.51 mm. Oral sucker terminal, round to oval, 0.07 - 0.09 mm x 0.08 - 0.09 mm. Prepharynx present. Pharynx oval, 0.02 -0.03 mm x 0.03 - 0.07 mm. Intestinal caeca absent. Ventral sucker round, 0.09 - 0.11 mm 0.08 - 0.11 mm, larger than oral sucker, pre-equatorial, circular. Pseudosuckers slightly compressed, close to pharynx. Holdfast organ ‘U’ shaped, bilobed, posterior to ventral sucker, lodges a distinct cavity, 0.17 mm x 0.13 mm. Holdfast gland ‘W’ shaped, consists of deeply stained cell mass, which partly covers posterior border of holdfast organ. Gonad is represented by one small, dark stained, elongate-oval mass of cells, located posterior to holdfast organ, at hind region of body. Details of reserve excretory system could not be studied due to small number of specimens collected.

Discussion

The present larva closely resembles with T. sophoriensis, Tetracotyle metacercaria, T. glossogobii, T. lucknowensis, T. singhi, T. tandani, T. lymnaei, T. gyanpurensis, T. pandei, and T. fotedari in number of genital rudiments and shape of body but differs from them in shape of holdfast organ and holdfast gland. It chiefly differs from T. glossogobii, T. lucknowensis, T. 41

singhi, T. lymnaei, and T. pandei in position of holdfast organ and holdfast gland, from T. sophoriensis, T. tandani and T. gyanpurensis in shape and position of pseudosuckers and from T. fotedari in ratio of suckers. Among the species described from foreign land, it comes closer to Tetracotyle of Cotyluris communis, Tetracotyle of Apatemon pellucidus, Tetracotyle of Apatemon fuligulae, Tetracotyle biwaensis and Tetracotyle tahoensis in ratio of suckers and presence of intestinal caeca but differs in shape of holdfast organ and holdfast gland. It further differs from Tetracotyle of Apatemon pellucidus and Tetracotyle of Apatemon fuligulae in number of genital rudiments and from Tetracotyle of Cotyluris communis, Tetracotyle biwaensis and Tetracotyle tahoensis in the presence of prepharynx and ratio of body. Therefore, the larva is regarded a new species and named Tetracotyle saiensis n. sp. after the locality where from the host was procured.

0.2mm 0.2mm

Fig.25 Fig.26

Tetracotyle saiensis n. sp. Fig.25 encysted metacercaria Fig.26 metacercaria

Tetracotyle daliganjensis n. sp. (Fig. 27 - 28)

Host: Xenentodon cancila (Ham.) Location: Liver Locality: Daliganj fish market, Lucknow No. of host examined: 15 No. of host found infected: 03 No. of metacercariae collected: 05

Cyst (Fig.27) elongate-oval, 0.80 - 0.92 mm x 0.25 - 0.40 mm, devoid of pigments. Body (Fig.28) large, oval, elongated, conical anteriorly, aspinose, thick walled, 1.35 - 1.54 mm x 0.30 - 0.52 mm. Fore body and hind body not clearly marked off. Oral sucker terminal, elongate-oval, 0.09 - 0.11 mm x 0.04 - 0.09 mm. Pseudosuckers well developed, forming a false collar, 0.12 - 0.14 mm x 0.09 - 0.12 mm. Prepharynx absent. Pharynx elongate-oval, well developed, muscular, 0.03 - 0.05 mm x 0.02 – 0.04 mm. Oesophagus absent. Intestinal caeca extending up to holdfast organ. Ventral sucker larger than oral sucker, equatorial, 0.10 - 0.14 mm x 0.09 - 0.12 mm. Hold fast organ bilobed, 0.29 - 0.32 mm x 0.11 - 0.13 mm, located behind ventral sucker, post equatorial, transversely elongated, with a median slit, bowl shaped. Hold fast gland ‘U’ shaped, protrusible. An small, 0.03 - 0.05 mm x 0.02 - 0.04 mm, dark stained, cell mass is rudimentary gonad located at hind end of body. Excretory bladder rounded, with small opening. Reserve excretory system detail could not be studied.

Discussion

The present larva closely resembles with T. ranae, Tetracotyle metacercaria, T. xenentodoni, T. glossogobii, T. lali, T. singhi, T. bufoi, T. aglandulata, T. kawi and T. satyapali in shape of body, position of 44

pseudosucker but differs from them in shape of holdfast organ and holdfast gland. It differs from T. xenentodoni, T. bufoi, and T. aglandulata in ratio of suckers, from T. glossogobii in number of genital rudiments, from T. ranae, and T. lali, in position of holdfast organ and holdfast gland, from T. kawi in absence of oesophagus and from T. satyapali in presence of intestinal caeca.

Among the species described from foreign land, it comes closer to Tetracotyle of Cotyluris communis, Tetracotyle of Apatemon fuligulae, Tetracotyle of Apatemon pellucidus, Tetracotyle biwaensis and Tetracotyle tahoensis in ratio of suckers but differs in shape of holdfast organ and holdfast gland. It further differs from Tetracotyle of Cotyluris communis, Tetracotyle of Apatemon fuligulae in number of genital rudiments, from Tetracotyle of Apatemon pellucidus and Tetracotyle biwaensis in position of holdfast organ and holdfast gland and absence of oesophagus and from Tetracotyle tahoensis in absence of pharynx. Therefore, the larva is regarded a new species and named Tetracotyle daliganjensis n. sp. after the locality wherefrom the host was procured.

0.2mm 0.2mm

Fig.27 Fig.28

Tetracotyle daliganjensis n. sp. Fig.27 encysted metacercaria Fig.28 metacercaria

Tetracotyle megapseudosuckerai n. sp. (Fig. 29 - 30)

Host: Mastacembelus armatus (Lacep.) Location: Liver Locality: Daliganj No. of host examined: 20 No. of host found infected: 10 No. of metacercariae collected: 09

Cyst (Fig.29) oval, 0.85 - 0.95 mm x 0.45 - 0.50 mm, single layered, devoid of pigments. Body (Fig.30) aspinose, foliaceous, divided into fore and hind body. Fore body 0.65 - 0.72 mm x 0.52 - 0.63 mm and hind body 0.20 - 0.28 mm x 0.18 - 0.36 mm. Oral sucker terminal, 0.03 mm - 0.06 mm x 0.04 - 0.06 mm. Prepharynx absent. Pharynx subglobular or elongate-oval, 0.02- 0.04 mm x 0.02 - 0.03 mm. Oesophagus short, 0.20 mm x 0.01 mm, intestinal caeca extending upto ventral sucker. Pseudosuckers large, well developed and located antero-laterally, 0.20 - 0.26 mm x 0.14 - 0.18 mm. Ventral sucker pre- equatorial, circular, larger than oral sucker, 0.10 - 0.13 mm 0.10 - 0.13 mm. Hold fast organ situated behind ventral sucker, 0.45 - 0.52 mm x 0.15 - 0.21 mm. Hold fast gland located close to hold fast organ. Two dark stained, tandem cell masses are rudimentary gonads, in hind end of body. Anterior mass may be presumptive testes while posterior mass may be ovary. Anterior mass 0.03 - 0.05 mm x 0.09 - 0.11 mm, elongate-oval, behind holdfast organ, and posterior mass 0.02 - 0.04 mm x 0.08 - 0.10 mm. Excretory bladder V- shaped. Excretory pore terminal. Reserve excretory system details could not be studied.

Discussion

The present larva closely resembles with T. ranae, T. sophorensis, T. indicus, T. metacercariae, T. szidati, T. muscularis, T. glossogobii, T. lali, T. lucknowensis, T. singhi, T. baughi, T. tandani, T. lymnaei, T. gyanpurensis, T. chauhani, T. pandei, T. srivastavai, T. satendri, T.kawi, T. kalyani and T. satyapali in ratio of suckers but differs from them in shape of hold fast organ and hold fast gland. It further differs from T. ranae, T. sophorensis, T. indicus, T. szidati, T. muscularis, T. glossogobii, T. lali, T. singhi, T. baughi, T. tandani, T. chauhani, T. srivastavai, T.kawi, T. kalyani and T. satyapali in number of genital rudiments, from T. metacercariae and T. lymnaei in shape of body and position of hold fast organ and hold fast gland and from T. lucknowensis, T. gyanpurensis, T. pandei and T. satendri in ratio of fore and hind body.

Among the species described from foreign land, it comes closer to Tetracotyle diminuta, Tetracotyle of Cotyluris communis, Tetracotyle of Apatemon fuligulae, Tetracotyle of Apatemon pellucidus, Tetracotyle biwaensis and Tetracotyle tahoensis in presence of oesophagus and intestinal caeca but differs in number of genital rudiments. It further differs from T. diminuta in ratio of suckers, from Tetracotyle of Cotyluris communis in presence of pharynx, from Tetracotyle of Apatemon fuligulae and Tetracotyle of Apatemon pellucidus by position and shape of hold fast organ and hold fast gland and from Tetracotyle biwaensis and Tetracotyle tahoensis in ratio of body. Therefore, the larva is regarded a new species and named Tetracotyle megapseudosuckerai n. sp., due to presence of large pseudosuckers.

0.2mm

Fig.29 Fig.30

Tetracotyle megapseudosuckerai n. sp. Fig.29 encysted metacercaria Fig.30 metacercaria 49

Tetracotyle multilobulata n. sp. (Fig. 31 - 32)

Host: Macrognethus aculeatus (Bloch) Location: Cranium Locality: Daliganj, river Gomati No. of host examined: 10 No. of host found infected: 01 No. of metacercariae collected: 03

Cyst (Fig.31) oval, 0.60 - 0.75 mm x 0.40 - 0.48 mm, devoid of pigments. Body (Fig.32) aspinose, foliaceous and not clearly divided into fore and hind body, 0.75 – 0.82 mm x 0.58 - 0.63 mm. Oral sucker sub-terminal, elongate-oval, 0.08 - 0.10 mm x 0.10 - 0.13 mm. Ventral sucker pre- equatorial, 0.08 - 0.10 mm x 0.10 - 0.13 mm. Prepharynx absent. Pharynx present, subglobular, prominent, 0.04 - 0.06 mm x 0.02 - 0.05 mm.. Pseudosuckers well developed and located antero-laterally, 0.29 - 0.32 mm x 0.11 - 0.17 mm, extending upto ventral sucker. Oesophagus long, intestinal caeca reaching upto holdfast organ. Hold fast organ situated behind ventral sucker, 0.45 - 0.55 mm x 0.26 - 0.30 mm. Hold fast gland not clearly visible. Genital rudiments represented by three stained cell masses. Two transversely elongated, masses may be presumptive testes while an antero-lateral round to oval mass ovary. Anterior mass 0.10 - 0.13 mm x 0.02 - 0.04 mm, antero- lateral mass 0.10 - 0.12 mm x 0.03 - 0.04 mm., and hind mass 0.01 - 0.02 mm x 0.01 – 0.02 mm. Excretory bladder ‘Y’ shaped. Excretory pore terminal. Reserve excretory system details could not be studied

Discussion

The present larva closely resembles with, T. sophorensis, T. ujjainensis, T. szidati, T. muscularis, T. glossogobii, T. baughi, T. tandani, T. 50

chauhani, T. simhai, T. fotedari, and T. kalyani in the number of genital rudiments but differs from them in shape of hold fast organ. It further differs from T. shophorensis and T. szidati in ratio of suckers and position of pseudosuckers, from T. ujjainensis, T. muscularis, T. glossogobii, T. baughi, T. tandani, T. simhai and T. fotedari in ratio of body and from T. chauhani and T. kalyani in position of hold fast organ.

Among the species described from foreign land, it comes closer to Tetracotyle diminuta, Tetracotyle of Cotyluris communis, Tetracotyle of Apatemon fuligulae, Tetracotyle of Apatemon pellucidus, Tetracotyle biwaensis and Tetracotyle tahoensis in presence of oesophagus and intestinal caeca but in differs in shape of hold fast organ. It further differs from Tetracotyle of Cotyluris communis, Tetracotyle of Apatemon fuligulae, Tetracotyle biwaensis and Tetracotyle tahoensis in number of genital of rudiments and ratio of suckers, from T. diminuta in ratio of body and position of pseudosuckers and from Tetracotyle of Apatemon pellucidus in presence of pharynx and position of hold fast organ. Therefore, the larva is regarded a new species and named Tetracotyle multilobulata n. sp. after the presence of multilobed holdfast organ.

0.2mm

Fig.31 Fig.32

Tetracotyle multilobulata n. sp. Fig.31 encysted metacercaria Fig.32 metacercaria

Tetracotyle varanasiensis n. sp. (Fig. 33 - 34)

Host: Macrognethus aculeatus (Bloch) Location: Gut Locality: Varanasi No. of host examined: 25 No. of host found infected: 15 No. of metacercariae collected: 17

Cyst (Fig.33) oval, single layered, pigmented, transparent, 0.68 - 0.85 mm x 0.35 - 0.55 mm. Body (Fig.34) aspinose, elongate-oval, large, divided into broadly oval fore body, 0.70 - 0.81 mm x 0.60 – 0.77 mm and small, conical, hind body, 0.20 - 0.25 mm x 0.23 - 0.41 mm. A collar is present at anterior region of body. Oral sucker terminal, round to oval, 0.08 - 0.10 mm x 0.04 - 0.09 mm. Prepharynx absent. Pharynx elongate-oval, 0.02 - 0.04 mm x 0.02 - 0.03 mm. Intestinal caeca not visible. Ventral sucker round to oval, encased by holdfast organ and holdfast gland, larger than oral sucker, 0.10 - 0.13 mm x 0.08 - 0.12 mm. Pseudosuckers well developed, elongate-oval, close to pharynx, 0.21 - 0.23 mm x 0.12 - 0.14 mm. Hold fast organ bilobed, 0.40 - 0.52 mm x 0.29 - 0.35 mm. Hold fast gland present. Rudimentry gonads two, masses of dark staining cells, posterior to hold fast gland. Anterior mass may be presumptive testis, 0.09 - 0.10 mm x 0.03 - 0.05 mm, while posterior mass 0.08 - 0.10 mm x 0.02 - 0.05 mm, ovary. Bursa copulatrix absent. Excretory bladder ‘V’ shaped, excretory pore terminal. Reserve excretory system details could not be studied.

Discussion

The present larva closely resembles with T. ranae, T. sophoriensis, T. indicus, T. szidati, T. muscularis, T. glossogobii, T. lali, T. lucknowensis, T. 53

singhi, T. baughi, T. tandani, T. lymnaei, T. gyanpurensis, T. chauhani, T. pandei, T. srivastavai, T. satendri, T. kawi, T. kalyani and T. satyapali in ratio of suckers but differs from them in shape of holdfast organ and holdfast gland. It chiefly differs from T. sophorensis, T. szidati, T. tandani, T. chauhani, T. srivastavi, T. kalyani and T. satyapali in number of genital rudiments, from T. szidati, T. muscularis and T. baughi in position of pseudosuckers, from T. ranae, T. indicus, T. lali, T. lucknowensis, T. singhiand and T. kawi in ratio of body and absence of oesophagus and intestinal caeca and from T. metacercariae in position of hold fast organ and hold fast gland Among the species described from foreign land, it comes closer to Tetracotyle of Cotylurus communis, Tetracotyle of Apatemon fuligulae, Tetracotyle of Apatemon pellucidus, Tetracotyle biwaensis and Tetracotyle tahoensis in ratio of sucker but differs in shape of hold fast organ and hold fast gland. It further differs from Tetracotyle of Cotylurus communis, Tetracotyle of Apatemon fuligulae and Tetracotyle of Apatemon pellucidus in number of genital rudiments and absence of osophagus and intestinal caeca, and from Tetracotyle biwaensis and Tetracotyle tahoensis in shape of body, ratio of body and position of ventral sucker. Therefore, the larva is regarded a new species and named Tetracotyle varanasiensis n. sp. after the the locality wherefrom the host was procured.

0.2mm 0.2mm

Fig.33 Fig.34

Tetracotyle varanasiensis n. sp. Fig.33 encysted metacercaria Fig.34 metacercaria

Tetracotyle trilobulata n. sp. (Fig. 35 - 36)

Host: Mastacembelus armatus (Lacep.) Location: Liver Locality: Daliganj No. of host examined: 10 No. of host found infected: 05 No. of metacercariae collected: 10

Cyst (Fig.35) round to oval, 0.62 - 0.74 mm x 0.41 - 0.55 mm. Body (Fig.36) aspinose, oval, not clearly divided into fore and hind body, 0.80 - 0.96 mm x 0.60 - 0.73 mm. Oral sucker sub-terminal, round to oval, 0.06 - 0.08 mm x 0.05 - 0.07 mm. Ventral sucker pre-equatorial, larger than oral sucker, round to oval, 0.08 - 0.11 mm x 0.09 - 0.12 mm. Pseudosuckers well developed. Pre-pharynx absent. Pharynx sub-globular, 0.03 - 0.04 mm x 0.02 - 0.03 mm. Hold fast organ and holdfast gland present. Gonadal rudiment represented by a tiny mass of dark staining cells, located at posterior end of body. Bursa copulatrix not observed. Excretory bladder ‘V’ shaped. Excretory pore terminal. Details of reserve excretory system could not be observed.

Discussion

The present larva closely resembles with T. ranae, T. indicus, T. lali, T. singhi, T. aglandulata, T. srivastavai, T. ramalingi, T. kawi T. satyapali and T. bufoi in the number of genital rudiment but differs from them in shape of hold fast organ and hold fast gland. It further differs from T. ranae and T. kawi in shape of body, absence of intestinal caeca and position of pseudosuckers, from T. indicus, T. srivastavai and T. ramalingi in ratio of body, from T. lali in absence of oesophagus, from T. xenentodoni, T. 56

aglandulata and T. bufoi in ratio of suckers, from T. singhi in absence of prepharynx and from T. satyapali in position of hold fast organ and hold fast gland. Among the species described from foreign land, it comes closer to, Tetracotyle of Cotyluris communis, Tetracotyle of Apatemon fuligulae, Tetracotyle of Apatemon pellucidus, Tetracotyle biwaensis and Tetracotyle tahoensis in ratio of suckers but differs in absence of oesophagus and intestinal caeca. It further differs from Tetracotyle of Cotyluris communis in ratio of body and position of pseudosuckers, from Tetracotyle of Apatemon fuligulae in position of hold fast organ and hold fast gland and absence of pre pharynx, from Tetracotyle of Apatemon pellucidus in number of genital rudiments and presence of pharynx and from Tetracotyle biwaensis and Tetracotyle tahoensis in shape of hold fast organ and hold fast gland. Therefore, the larva is regarded a new species and named Tetracotyle trilobulata n. sp. because of the presence of a trilobed holdfast organ.

0.2mm 0.2mm

Fig.35 Fig.36

Tetracotyle trilobulata n. sp. Fig.35 encysted metacercaria Fig.36 metacercaria

Metacercaria of Bucephalopsis garuai Verma, 1936 (Fig. 37 - 38)

Host: Clupisoma garua (Ham.) Location: Gill Locality: Chapra (Bihar) No. of host examined: 10 No. of host found infected: 03 No. of metacercariae collected: 05

Cyst (Fig.37) small, round to oval, thin, delicate, easily breakable, 0.20 - 0.22 mm x 0.08 - 0.11 mm. Body (Fig.38) cyclindrical or spindle-shaped, aspinose, with broader posterior end, 0.40 - 0.45 mm x 0.22 - 0.26 mm. Broader apical sucker well developed, sub-terminal, 0.09 - 0.11 mm x 0.10 - 0.13 mm,. Pharynx well developed, spherical, post-equatorial, 0.01 - 0.02 mm x 0.01 - 0.02 mm. Oesophagus not prominent. Gut sac-like, long right side of pharynx, 0.60 - 0.70 mm x 0.55 - 0.62 mm. Reproductive organs well marked. Ovary 0.01 - 0.02 mm x 0.02 - 0.03 mm, small, globular, near left margin, at junction of middle of body, sinistral, at level of pharynx, pre-testicular, followed by a small darkly stained mass may be shell gland. Testes two, round to oval, left to pharynx, one behind each other, 0.02 - 0.03 mm x 0.01 - 0.02 mm each. Uterus slender, tube, darkly stained, running in sinuous course towards right side of body and along cirrus sac, opening into genital tongue. Genital tongue long, slender. Vitellaria not yet developed. Vasa efferentia arise from testes and form a common duct, vas deferens opens into cirrus. Cirrus sac long, 0.15 - 0.17 mm x 0.02 - 0.03 mm, near posterior end of body, along left margin, enclosing vesicula seminalis and a double layered broad pars prostatica, surrounded by prostate gland cells. Excretory bladder very large, saccular, occupying space between pharynx to cirrus sac. Excretory pore terminal and opens into genital pore.

Discussion

To the best of my knowledge, the following species of metacercariae are known under the genus Bucephalopsis (Diesing, 1855) Yamaguti, 1958, from India viz. metacercaria of B. garuai Verma, 1936, metacercaria of B. fusiformis Verma, 1936, B. devi Sinha, 1964, Bucephalopsis metacercaria Prasad & Sinha, 1964, Bucephalopsis metacercaria Pande and Rai, 1964, B. pentaglandulata Chakrabarti, 1968, B. hexaglandulata Pandey, 1969, B. multiglandulata Pandey, 1969, B. oxygasteri Pandey, 1969 and B. linguiformis Chakrabarti and Baugh, 1974. Of these, the present larva closely resembles with metacercaria of B. garuai described by Pande, Chauhan and Arora in 1968, from musculature and viscera of Bufo andersonii (Boulinger), Aspidoparia morar (Ham.) and Barilius ovazardi (Day) from Jamuna river, Mathura. This is a new host and locality record. The appended table shows the measurements of various body organ described by Pande et al., and present larva.

Table

Name of Metacercaria of Bucephalopsis Present specimens organs garuai Verma, 1936 Cyst 0.23 mm x 0.38 mm 0.20-0.22 mm x 0.08-0.11 mm Body 0.63-0.88 mm x 0.23-0.35 mm 0.40-0.45 mm x 0.22-0.26 mm Ap. sucker ? 0.09-0.11 mm x 0.10-0.13 mm Pharynx 0.04 mm x 0.06 mm 0.01-0.02 mm x 0.01-0.02 mm Gut ? 0.60-0.70 mm x 0.55-0.62 mm Ovary 0.020 mm x 0.022 mm 0.01-0.02 mm x 0.02-0.03 mm Testes 0.02 mm x 0.027 mm 0.02-0.03 mm x 0.02-0.03 mm Cirrus 0.07-0.14 mm x 0.02-0.03 mm 0.15-0.17 mm x 0.02-0.03 mm

0.1mm 0.1mm

Fig.37 Fig.38

Metacercaria of Bucephalopsis garuai Verma, 1936 Fig.37 encysted metacercaria Fig.38 metacercaria 61

Metacercaria of B. linguiformis Chakrabarti and Baugh, 1974 (Fig. 39 - 40)

Host: Clarias batrachus (Linn.) Location: Liver Locality: Kaisarbagh, Fish market (Lucknow) No. of host examined: 10 No. of host found infected: 01 No. of metacercariae collected: 01

Cyst (Fig.39) small, oval, thin walled, membranous, 0.25 - 0.30 mm x 0.08 - 0.10 mm. Body (Fig.40) aspinose, elongated, with rounded ends, 0.40 - 0.42 mm x 0.14 - 0.16 mm. Apical sucker terminal, round to oval, 0.06 - 0.07 mm x 0.05 - 0.06 mm. Mouth equatorial. Pharynx round to oval, 0.03 mm x 0.04 mm. Oesophagus short, 0.004 - 0.006 mm x 0.005 - 0.006 mm. Intestine saccular, containing mass of round bodies of various sizes, 0.06 - 0.07 mm x 0.04 - 0.05 mm. Gonads fairly well differentiated, in posterior half of body. Testes two, small, round, equal masses, behind pharynx, 0.01 - 0.03 mm x 0.01 - 0.02 mm each. Ovary small cell mass, pretesticular, below pharynx, 0.008 - 0.01 mm x 0.005 - 0.01 mm. Cirrus sac cylindrical, on right side of body, narrow anteriorly and broader towards posterior region, enclosing long tubular seminal vesicle, with incompletely developed pars prostatica and ejaculatory duct, 0.12 - 0.20 mm x 0.05 - 0.10 mm. Vitelline follicles not yet developed. Excretory bladder elongated, tubular, roughly sigmoid, raching upto oral sucker, with excretory granules, opening outside by terminal excretory pore.

Discussion

The present larva closely resembles with metacercaria of B. linguiformis Chakrabarti and Baugh, 1974. Chakrabarti and Baugh (1974) 62

described the larva from musculature and body cavity of Esomus danricus (Ham) and Nemachilus botia (Ham.) from river Gomti, Lucknow. This is a new host and locality record. The appended table shows the measurements of various body organs described by Chakrabarti and Baugh (1974) and present specimens.

Table

Name of B. linguiformis metacercaria Present specimens organs Chakrabarti and Baugh, 1974 Cyst 0.22-0.30 mm x 0.18-0.24 mm 0.25 - 0.30 mm x 0.08 - 0.10 mm Body 0.64-0.38 mm x 0.18-0.26 mm 0.40 - 0.42 mm x 0.14 - 0.16 mm Ap. sucker 0.08 mm x 0.14 mm 0.06 - 0.07 mm x 0.05 - 0.06 mm Pharynx 0.04 mm x 0.07 mm 0.03 mm x 0.04 mm Oesophagus ? 0.004 - 0.006 mm x 0.005 - 0.006 mm Gut ? 0.06 - 0.07 mm x 0.04 - 0.05 mm Ovary ? 0.008 - 0.01 mm x 0.005 - 0.01 mm Testes 0.03-0.04 mm x 0.05-0.06 mm 0.01 - 0.03 mm x 0.01 - 0.02 mm Cirrus 0.18 mm x 0.24 mm 0.10 - 0.20 mm x 0.05 - 0.10 mm

0.1mm 0.1mm

Fig.39 Fig.40

Metacercaria of B. linguiformis Chakrabarti and Baugh, 1974 Fig.39 encysted metacercaria Fig.40 metacercaria

Metacercaria of Orchipedum Braun, 1901 (Fig. 41 - 42)

Host: Channa punctatus (Bloch) Location: Body cavity Locality: Fish market, Sector-3 (Lucknow) No. of host examined: 10 No. of host found infected: 01 No. of metacercariae collected: 03

Cyst (Fig.41) small, oval, thin walled, 1.25 - 1.30 mm x 0.55 - 0.68 mm. Body (Fig.42) aspinose, linguiform, with rounded ends, 2.05 - 2.10 mm x 0.60 - 0.81 mm. Suckers well developed. Oral sucker terminal, almost circular, 0.18 - 0.23 mm x 0.19 - 0.24 mm. Prepharynx absent. Pharynx small and opens into short oesophagus, 0.03 - 0.04 mm x 0.05 - 0.06 mm., ending into a swollen bulb, surrounded by a large number of unicellular glands, at intestinal bifurcation. Intestinal caeca have smooth margins, broad anteriorly but narrow posteriorly, extending upto hind end of body. Ventral sucker round to oval, very large, 0.65 - 0.71 mm x 0.60 - 0.65 mm. Genital rudiments not fully developed. Testes represented by a number of darkly stained, round cell mass in posterior region of body. Their number could not be ascertained. Ovary represented by round mass, pre-testicular, 0.04 - 0.06 mm x 0.06 - 0.08 mm, located behind ventral sucker. Vitelline follicles, though not yet well developed, foreshadowed by small granules in the lateral margin behind ventral sucker. Excretory bladder long, tubular and opens outside by a subterminal excretory pore.

Discussion

The larva has a conical forebody and backwardly tapered hind body, ventral sucker larger than oral sucker. No prepharynx, oesophagus very short 65

or practically absent. Caeca reaching in posterior region of body, testes arranged in one row at level of caecal ends, genital pore immediately behind intestinal bifurcation., ovary rounded, on one side of median line, immediately infront of testicular area, receptaculum seminis small, posterior to overy. Uterus winding mostly in intercaecal field, between tetstes and genital pore, therefore it appears to be a metacercariae of avian trematode Orchipedum Braun, 1901. To the best of my knowledge, this is the first record of the present metacercaria of above genus from India. It is probable that some piscivorus bird feeds an infected fishes and the metacercariae develop into adults.

0.5mm

0.1mm

Fig.41 Fig.42

Metacercaria of Orchipidum Braun, 1901 Fig.41 encysted metacercaria Fig.42 metacercaria

Metacercaria of Opisthorchis elongatus Agrawal, 1975 (Fig. 43 - 44)

Host: Esomus danricus (Ham.) Location: Gill Locality: Aquarium shops (Lucknow) No. of host examined: 10 No. of host found infected: 03 No. of metacercariae collected: 05

Cyst (Fig.43) spherical or ovoidal, transparent, thick, double layered, 0.40 - 0.44 mm x 0.10 - 0.15 mm. Body (Fig.44) elongate-oval, leaf like, aspinose, 0.55 - 0.62 mm x 0.15 - 0.21 mm. Oral sucker terminal, circular, 0.08 - 0.10 - 0.08 - 0.10 mm. Prepharynx absent. Pharynx well developed, globular, 0.05 - 0.07 mm x 0.04 - 0.06 mm. Oesophagus long, 0.10 mm - 0.13 mm., divides near ventral sucker into short, divergent caeca, reaching upto posterior testis. Ventral sucker postequatorial, smaller than oral sucker, circular, 0.04 - 0.06 mm x 0.04 - 0.06 mm. Gonads well developed. Testes two, obliquely placed, round to oval, anterior testis close to ventral sucker, circular, 0.02 - 0.03 mm x 0.02 - 0.03 mm., while posterior testis close to end of intestinal caeca, 0.02 - 0.03 mm x 0.01 - 0.02 mm. Ovary small, close to posterior margin of ventral sucker, 0.01 - 0.02 mm - 0.01 - 0.02 mm. Thick dark staining band of cells, representing cirrus sac, runs from anterior margin of ventral sucker and ends infront of ovarian rudiments. Excrtetory bladder well developed, elongate oval and filled with excretory granules. Excretory pore terminal.

Discussion

To the best of my knowledge, following species of metacercariae are known under the genus Opisthorchis metacercaria R. Blanchard, 1895, from 68

India viz. O. caninus Rai and Pande, 1965 and O. elongatus Agrawal, 1975. Of these, the present larva closely resembles with O. elongatus. It is briefly recorded here. Agrawal, 1975 described this larva from mantle wall of Vivipara bengalensis (Linn.) from Lucknow. This is a new host and locality record of the worm. The appended table shows the measurements of various body organs, described by Agrawal, 1975.

Table

Name of Metacercaria of Opisthorchis Present species organs elongatus Agrawal, 1975 Cyst ? 0.40-0.44 mm x 0.10-0.15 mm Body ? 0.55-0.62 mm x 0.15-0.21 mm Oral sucker ? 0.08-0.10 mm x 0.08-0.10 mm Pharynx ? 0.05-0.07 mm x 0.04-0.06 mm Oesophagus ? 0.10 mm x 0.13 mm V. Sucker ? 0.04-0.06 mm x 0.04-0.06 mm Ovary ? 0.01-0.02 mm x 0.01-0.02 mm Testes ? 0.02-0.03 mm x 0.01-0.02 mm.

0.2mm 0.2mm

Fig.43 Fig.44

Metacercaria of Opisthorchis elongates Agrawal, 1975 Fig.43 encysted metacercaria Fig.44 metacercaria 70

Plagiorchiid metacercaria (Fig. 45 - 46)

Host: Channa punctatus (Bloch) Location: Body cavity Locality: Fish market, Sector: 3 (Lucknow) No. of host examined: 10 No. of host found infected: 01 No. of metacercariae collected: 05

Cyst (Fig.45) round to oval, thick, 0.78 - 0.82 mm x 0.42 - 0.49 mm. Body (Fig.46) annulated, cigar shaped, with narrow anterior and broad posterior ends, 1.10 - 1.22 mm x 0.45 - 0.63 mm. Prepharynx absent. Pharynx prominent, large, muscular, round to oval, 0.10 - 0.13 mm x 0.08 - 0.11 mm. Oral sucker terminal, well marked, round to oval, thick, muscular, 0.14 - 0.19 mm x 0.20 - 0.24 mm. Oesophagus short, leading into two intestinal caeca. Intestinal caeca long, reaching upt hind end of body. Ventral sucker in middle of body, almost equal to oral sucker, 0.13 - 0.18 mm x 0.18 - 0.22 mm. A mass of unicellular glands scattered in space between intestinal bifurcation and ventral sucker. Genital primordia consist of 2 small clusters of cells, diagonally placed, representing testicular rudiments, intercaecally. Ovarian rudiment pre-testicular behind ventral sucker. Cirrus sac represented by a mass of cells near anterior margin of anterior margin of ventral sucker. Excretory vesicle ‘Y’ shaped and contains excretory granules. Excretory pore terminal.

Discussion

Since the metacercaria has an elongate oval body, well developed suckers, well developed pharynx, intestinal caeca extending upto hind region of body, prominent genital rudiment, ovary pretesticular, testes one behind 71

other, cirrus sac around ventral sucker and ‘Y’ shaped excretory bladder, the larva belongs to Plagiorchiid group of trematodes.

To the best of my knowledge the following Plagiorchiid metacercariae are known from India viz. P. metacercaria Rai and Pande, 1965, Type II Matta and Pande, 1966, Type III Matta and Pande, 1966, Type IV Matta and Pande, 1966, Type V Matta and Pande, 1966, Type VI Matta and Pande, 1966, Type VII Matta and Pande, 1966, Type VIII Matta and Pande, 1966 and Plagiorchiid type IX Pandey and Tiwari, 1983. However, without performing feeding experiments, it is difficult to acertain the adult trematode of this metacercaria.

0.2mm 0.2mm

Fig.45 Fig.46

Plagiorchiid metacercaria Fig.45 encysted metacercaria Fig.46 metacercaria

Metacercaria of Ommatobrephus Mehra, 1928 (Fig. 47- 48)

Host: Colisa fasciatus (Bloch. & Schn.) Location: Body cavity/Gut Locality: Sector: 3 (Lucknow) No. of host examined: 10 No. of host found infected: 01 No. of metacercariae collected: 01

Cyst (Fig.47) round to oval, thin layered, 1.27 - 1.30 mm x 0.45 - 0.55 mm. Body (Fig.48) aspinose, with narrow, rounded anterior and broad posterior ends, 2.60 - 2.67 mm x 0.70 - 0.98 mm. Oral sucker terminal, almost circular, small, 0.22 - 0.25 mm x 0.21-0.24 mm. Prepharynx absent. Pharynx large, 0.16 - 0.18 mm x 0.18 - 0.21 mm. Oesophagus long, thick, muscular, 0.35 - 0.40 mm. Intestinal Caeca smooth, extend upto hind end, reaching upto level of testes. Ventral sucker equatorial, round to oval, large, 0.50 - 0.56 mm x 0.60 - 0.63 mm. Gonads well developed, intercaecal, represented by three darkly stained cell masses, at hind end of intestinal bifurcation. Testes elongated, lobed, parallel, laterally placed, 0.11 - 0.16 mm x 0.10 - 0.15 mm. Ovary small, pretesticular, round to oval, 0.05 - 0.07 mm x 0.06 - 0.08 mm. Uterus coiled, tubular. Cirrus sac elongate-oval, muscular, between intestinal bifurcation, occupied by a bipartite vesicula seminalis, 0.25 - 0.30 mm x 0.05 - 0.09 mm. Excretory bladder triangular, filled with rounded blackish excretory granules.

Discussion

The general topography of organ clearly shows the relationship of the larva with the genus Ommatobrephus Mehra, 1928, particularly in having fusiform to elongated pyriform, unspined, flattened body, large ventral sucker, 74

pre-equatorial, oral sucker small, pharynx comparatively large, esophagus moderately long, caeca reaching to posterior extremity, testes symmetrical, at posterior extremity, cirrus sac pyriform, entirely pre-acetabular, seminal vesicle convoluted, genital pore median, post-bifurcal, ovary a little to one side of median line, just pre-testicular. Therefore, it represents the metacercaria of genus Ommatobrephus Nicoll, 1914. To the best of my knowledge, so far no metacercaria of above genus has been reported in India. This appears to be the first record of the metacercaria of Ommatobrephus. Adult species are known from snakes and Varanus viz. O. lobatum Mehra, 1929, O. folium Thapar and Ali, 1929, O. lobatum najii Mehra, 1931, O. nicolli Gupta, 1954, O. medaacetabulus Simha, 1958, O. chauhani Dwivedi, 1968, O. bengalensis Mukherjee and Ghosh, O. minutum Dwivedi and Chauhan, 1969 and O. Piscatori Dandotia, 1972. However, without performing feeding experiments it is difficult to acertain to which adult species, this metacercaria belongs.

0.2mm 0.1mm

Fig.47 Fig.48

Metacercaria of Ommatobrephus Mehra, 1928 Fig.47 encysted metacercaria Fig.48 metacercaria

Metacercaria of Isoparachis hypselobagri (Billet, 1898) Odhner, 1927

(Fig. 49)

Host: Wallago attu (Bloch & Schn.) Location: Body cavity Locality: Kanpur fish market No. of host examined: 10 No. of host found infected: 03 No. of metacercariae collected: 04

Cyst absent. Body (Fig.49) aspinose, thick, elongated, anterior end conical, posterior end broad, 10.70 - 10.80 mm x 2.40 - 4.80 mm. Oral sucker round to oval, terminal, muscular, 0.80 - 0.90 mm x 0.75 - 0.80 mm. Prepharynx absent. Pharynx well developed, 0.40 - 0.45 mm x 0.38 - 0.40 mm. Oesophagus very short, 0.10 - 0.12 mm x 0.08 - 0.10 mm. Intestinal caeca serrated, broad, node like structures, at turning point, and appears yellow, extending upto posterior extremity. Ventral sucker pre-equatorial, circular, slightly smaller than oral sucker, 0.75 mm - 0.80 mm. Gonads well developed. Testes appear as two small, circular bodies, located at sides of ventral sucker in intercaecal field, 0.42 - 0.50 mm x 0.42 - 0.55 mm. Vasa efferentia, arising from testes unite infront of ventral sucker, to form a short vas deferens which continued into a narrow vesicula seminalis. Genital pore median. Ovary at hind end of body, having fine branches, each ending in a vesicle containing elliptical nodes. A transversely elongated structure represents uterus. Vitellaria incipient. Excretory bladder elongated, intercalated between terminal parts of intestinal caeca and leads outside by a terminal excretory pore.

Discussion To the best of my knowledge, the only one metacercarial species of Isoparachis hypselobagri (Billet, 1898) Odhner, 1927 is so far known, to which the worms under study closely resemble. It has already been described in detail by Pandey, (1969). It is, therefore, recorded here. This is a new locality record of the parasite. The appended table shows a comparision of the measurements of various body organ, described by Pandey (1969) and present larva.

Table

Name of Metacercaria of Isopararchis hypselobagri Present species organs (Billet, 1898) Odhner, 1927

Body 1.07 mm x 0.7 mm 10.70-10.80 mm x 2.40-4.80 mm Oral sucker 0.09-0.36 mm x 0.12-0.31 mm 0.80-0.90 mm x 0.75-0.80 mm Pharynx 0.08 mm x 0.09 mm 0.40-0.45 mm x 0.38-0.40 mm Oesophagus ? 0.10-0.12 mm x 0.08-0.10 mm V. Sucker 0.43 mm x 0.43 mm 0.75 mm x 0.80 mm Testes ? 0.42-0.50 mm x 0.42-0.55 mm

0.5mm

Fig.49 Metacercaria of Isoparachis hypselobagri (Billet, 1898) Odhner, 1927

Metacercaria of Genarchopsis goppo Ozaki, 1925 (Fig. 50 - 51)

Host: Channa punctatus (Bloch) Location: Gut Locality: Itwara fish market (Bhopal) No. of host examined: 10 No. of host found infected: 06 No. of metacercariae collected: 14

Cyst (Fig.50) round to oval or elliptical, thin, transparent, 0.70 - 0.85 mm x 0.20 - 0.25 mm. Body (Fig,.51) pear shaped or elliptical, 0.95 - 1.02 mm x 0.35 - 0.50 mm, broad anteriorly and narrow posteriorly. Oral sucker subterminal, round to oval, 0.18 - 0.21 mm x 0.17 - 0.20 mm. Prephyrynx absent. Pharynx well developed, muscular, leading into oesophagus, 0.05 - 0.07 mm x 0.08 - 0.10 mm. Oesophagus short, 0.01 - 0.03 mm x 0.02 - 0.03 mm. Intestinal caeca form a shoulder, reaching upto hind region of body, in pretesticular region, wall of caeca not smooth but have bulb like margins, where they unite. Ventral sucker large, almost double the size of oral sucker, thick, muscular, postequatorial, 0.36 - 0.41 mm x 0.34 - 0.42 mm. Testes preovarian, laterally placed, behind ventral sucker, parallel to each other, elongate-oval, 0.06 - 0.08 mm x 0.04 - 0.05 mm and 0.07 - 0.09 mm x 0.03 - 0.05 mm respectively. Cirrus sac oval, pre acetabular, behind intestinal bifurcation, 0.08 - 0.10 mm x 0.04 - 0.06 mm. It contains a prominent oval vesicula seminalis, pars prostatica, surrounded by prostrate glands and a short ejaculatory duct. Ovary median, 0.04 - 0.06 mm x 0.02 - 0.03 mm, post testicular, elongate-oval. Vitellaria represented by two prominent dark stain cell masses, located at hind region of body. Excretory bladder elongated, Y shaped, wide, sinuous collecting ducts, one on each side and run in lateral field of body and unite anteriorly by a short transverse duct.

Discussion

Since the present metacercaria has fusiform body, subterminal oral sucker, well developed pharynx, oesophagus short, with or without a pouch posteriorly, caeca united together at or near posterior extremity, ventral sucker large, equatorial or postequatorial, testes diagonal, postacetabular, vesicula seminalis elongate, saccular or somewhat winding, far anterior to ventral sucker, pars prostatica distinct enclosed in a thin walled sac, ductus hermaphroditicus short. Therefore, it appears to be a metacercaria of trematode genus Genarchopsis goppo. This appears to be the first record of metacercaria from India. The adult worm is found in gut of frogs and fishes.

0.2mm 0.2mm

Fig.50 Fig.51

Metacercaria of Genarchopsis goppo Ozaki, 1925 Fig.50 encysted metacercaria Fig.51 metacercaria (drawn from live specimen)

Part-II

Adult

Bucephalus bhagalpurensis n. sp. (Fig.1)

Host: Mystus teengra (Ham.) Location: Gut Locality: River Saryu Bhagalpur, (Near Belthara, U. P.) No. of host examined: 08 No. of host found infected: 03 No. of worms collected: 17

Body (Fig.1) elongate, with minute backwardly directed spines, 1.4 - 3.2 mm x 0.28 - 0.68 mm. Anterior sucker 0.16 - 0.36 mm x 0.14 - 0.24 mm, with 4 - 6 well developed tentacles. Pharynx circular, 0.08 - 0.14 mm., leading into a mouth by short oesophagus. Oesophagus 0.01 - 0.02 mm. Intestine simple, ovoid sac, 0.24 - 0.4 mm x 0.12 - 0.2 mm, extending forward from pharynx to level of ovary. Testes tandem, on left side, separated by uterine coil. Anterior testis 0.14 - 0.25mm x 0.12 - 0.20 mm and posterior testes 0.12 - 0.21 mm x 0.10 - 0.15 mm. Cirrus sac elongate, oval, 0.16 - 0.18 mm x 0.03 - 0.10 mm, enclosing a small, ovoid, vesicula seminalis, 0.06 - 0.10 mm x 0.04 - 0.08 mm, pars prostatica, surrounded by prostate glands and a narrow ductus ejaculatorius and opening into a roughly triangular genital sinus at the base of tongue like structure, known as genital tongue or cone. Genital sinus subterminal, opens by genital pore. Ovary spherical, 0.10 - 0.18 mm x 0.08 - 0.12 mm. Vitellaria follicular, latral, extending from anterior margin of ovary to uterus. Uterus short, anteriorly, upto 1/4th of body and opens at genitial sinus by a short metraterm. Eggs 0.02 -0.03 mm x 0.01-0.02 mm, oval. Excretory bladder straight, tubular, extending from a little in front of anterior limit of vitellaria to excretory pore.

Discussion

To the best of my knowledge, following species are known under the genus Bucephalus Baer, 1826 in India viz. B. tridenticularia Verma, 1936; B. aoria Verma, 1936; B. jagannathai Verma, 1936; B. indicus Srivastava, 1938; B. gangeticus Srivastava, 1938; B. barina Srivastava, 1938; B. bagarius Srivastava, 1963; B. tritentacularis Srivastava, 1963; B. allahabadensis Srivastava, 1963; B. keni Agrawal and Agrawal, 1988; B. ranjeetpurai Maurya & Singh, 2005; B. fischthali Gupta and Tiwari, 1983; B. kanagurtai Gupta and Tiwari, 1983; B. thapari Gupta and Tiwari, 1983; B. yamagutii Gupta & Singh, 1985; B. octotentacularis Kakaji, 1969; B. gangi Maurya and Agrawal, 1992; B. dasashwamedhai Maurya and Agrawal, 1992; B. varanasiensis Maurya and Agrawal, 1992 and B. aori Maurya and Agrawal, 1992. The present form chiefly differs from all the above species in the position of pharynx in body, short cirrus sac and extension of vitellaria. The position of genital pore further seperates it from all the above species.

0.1mm

Fig.1 Bucephalus bhagalpurensis n. sp. (Ventral view)

Bucephalopsis karvei Bhalerao, 1937 (Fig. 2)

Host: A. coila (Ham), X. cancila (Ham.), E. vacha (Ham.) Location: Gut Locality: Fish market, Kanpur, Banaras and Barabanki No. of host examined: 50 No. of host found infected: 35 No. of worms collected: 40

Body (Fig.2) oval or pear-shaped, 1.68 - 2.5 mm x 0.82 - 1.2 mm rounded/blunt anterior end and a narrow, pointed, posterior end. Anterior sucker large 0.10 - 0.30 mm x 0.20 - 0.40 mm, ventral, situated at anterior end. Pharynx muscular, surrounding mouth, 0.08 - 0.10 mm x 0.10 - 0.12 mm. Intestine wide, 0.19 - 0.22 mm x 0.09 - 0.12 mm. Anterior testis 0.24 - 0.40 mm x 0.14 - 0.26 mm; posterior testis 0.35 - 0.60 mm x 0.24 - 0.30 mm. Ovary 0.26 - 0.32 mm x 0.26 - 0.30 mm, pear-shaped. Cirrus sac highly developed, 0.66 - 0.90 mm x 0.20- 0.30 mm, extending forward up to level of anterior testis and enclosing a vesicula seminalis, 0.20 - 0.30 mm x 0.14 - 0.20 mm. Pars prostatica surrounded by prostrate gland, 0.36 - 0.54 mm x 0.10 - 0.15 mm and a narrow ductus ejaculatorius, 0.08 - 0.10 mm x 0.10 - 0.15 mm. Genital tongue extends into funnel-shaped genital sinus, opening ventrally. Vitellaria large, rounded follicles, near anterior sucker, on the lateral sides of body, extending from ovary to that of the anterior sucker. Uterus extends forward up to the anterior limit of vitellaria. Excretory bladder tubular. Eggs light brown, 0.02 - 0.03 mm x 0.01 - 0.03 mm.

Discussion

B. karvei was described by Bhalerao (1937) from the intestine of Xenentodon cancilla, at Poona. Gupta (1958) also redescribed B. karve from the type host at Lucknow and recorded variations particularly in the disposition and size 87

of gonads and extention of vitellaria. Chauhan and Koche (1975) described B. gaurai from the intestine of Xenentodon cancilla (Ham.) at Jabalpur. It is characterized by the position of ovary and extension of vitelline follicle which are variable features. Therefore, I regard B. gaurai a synonym of B. karvei. The record of B. karvei from Kanpur, Varanasi and Barabanki appears to be new localities for the worm. The appended table shows a comparision of the measurements of various body organ, described by Bhalerao, 1937 and present larva.

Table

Name of Bucephalopsis karvei Bhalerao, Present specimens organs 1937 Body 0.50 - 0.96 mm x 0.27 - 0.57 mm 1.68 - 2.5 mm x 0.82 - 1.2 mm Apical sucker ? 0.10 - 0.30 mm x 0.20 - 0.40 mm Pharynx ? 0.08 - 0.10 mm x 0.10 - 0.12 mm Intestine ? 0.19 - 0.22 mm x 0.09 - 0.12 mm Ovary ? 0.26 - 0.32 mm x 0.26 - 0.3 mm Testes ? 0.24 - 0.60 mm x 0.14 - 0.30 mm Cirrus sac ? 0.66 - 0.90 mm x 0.20- 0.30 mm Ves.seminalis ? 0.20 - 0.30 mm x 0.14 - 0.20 mm P. prostatica ? 0.36 - 0.54 mm x 0.10 - 0.15 mm Ejacul. duct ? 0.08 - 0.10 mm x 0.10 - 0.15 mm Eggs 0.01 - 0.02 mm x 0.009 - 0.01 mm 0.02 - 0.03 mm x 0.01 - 0.03 mm

0.2mm

Fig.2 Bucephalopsis karvei Bhaleroa, 1937 (Ventral view)

Bucephalopsis fusiformis Verma, 1936 (Fig. 3)

Host: Eutropiicthys vacha (Ham.) Location: Gut Locality: Madhuban Tal No. of host examined: 15 No. of host found infected: 08 No. of worms/specimens collected: 15

Body (Fig.3) elngate, spinose, laterally compressed, rounded at both ends 1.48 - 1.78 mm x 0.25 - 0.30 mm. Anterior sucker terminal, 0.17 - 0.19 x 0.17 - 0.19 mm. Pharynx conspicuous, small, 0.03 - 0.05 mm x 0.05 - 0.08 mm. Oesophagus very short. Intestine wide, 0.21 - 0.23 mm x 0.10 - 0.13 mm. Testes round to oval, behind middle of body, anterior testis rounded or regular 0.10 - 0.15 mm x 0.06 - 0.10 mm; posterior testis, 0.08 - 0.13 mm x 0.05 - 0.09 mm. Cirrus sac long, from posterior testis to hind end of body, about one fourth as long as body, 0.46 - 0.59 mm x 0.05 - 0.09 mm. It includes a rounded vesicula seminalis, long pars prostatica surrounded by prostate glands and short ejaculatory duct, opening into a genital sinus by genital tongue. Ovary near right margin, rounded, 0.10 - 0.13 mm x 0.10 - 0.13 mm. Uterus with descending and ascending limbs, extending anterior to ovary and opens at genital sinus by a short metraterm. Genital pore at hind end of body. Vitellaria compact, follicles 14 - 16 in each lateral group, between ovary and anterior sucker, 0.03 mm x 0.03 mm. Eggs 0.01 - 0.02 mm x 0.084 - 0.01 mm.

Discussion

Verma (1936) described B. fusiformis from intestine of Eutropiicthys vacha (Ham.) at Allahabad. This appears to be a new record of the worm from Lucknow. The appended table shows a comparision of the measurements of various body organ, described by Verma ,1936 and present worm. 90

Table

Name of Bucephalopsis fusiformis Verma, Present specimens organs 1936 Body 1.24 - 2.52 mm x 0.39-0.84 mm 1.48 - 1.78 mm x 0.25 - 0.30 mm Apical sucker 0.17 mm x 0.24 mm 0.17 - 0.19 mm x 0.17 - 0.19 mm Pharynx 0.07 mm x 0.08 mm 0.03 - 0.05 mm x 0.05 - 0.08 mm Oesophagus ? 0.001 mm x 0.003 Ovary 0.14 mm x 0.21 mm 0.10 - 0.13 mm x 0.10 - 0.13 mm. Testes 0.16 - 0.29 mm x 0.15 - 0.26 mm 0.08 - 0.15 mm x 0.05 - 0.10 mm Cirrus sac 0.46 - 0.70 mm x 0.10 - 0.14 mm 0.46 - 0.59 mm x 0.05 - 0.09 mm Eggs 0.01 - 0.02 mm x 0.008 - 0.01 mm 0.01 - 0.02 mm x 0.084 - 0.01 mm

0.2mm

Fig.3 Bucephalopsis fusiformis Verma, 1936 (Ventral view)

Allocreadium itwarensis n. sp. (Fig. 4)

Host: Mastacembalus puncalus (Ham.) Location: Gut Locality: Itwara (Bhopal) No. of host examined: 07 No. of host found infected: 03 No. of worms collected: 08

Body (Fig.4) elongated, smooth, aspinose, 1.40 mm x .44 mm. Oral sucker terminal, more or less circular, 0.12 - 0.14 mm x 0.11 - 0.18 mm. Ventral sucker, pre-equatorial, larger than oral sucker, 0.16- 0.19 mm x 0.18 - 0.19mm. Prepharynx absent. Pharynx muscular, ovoid, 0.06 - 0.07 mm x 0.05 - 0.06 mm. Oesophagus short, 0.07 - 0.07 mm. Intestinal caeca simple, extending slightly behind posterior testis. Testes entire, oval, tandem, postequatorial, anterior testis 0.14 - 0.16 mm x 0.18 - 0.19 mm; posterior testis larger, close to anterior testis, 0.21 - 0.27 mm x 0.19 - 0.21 mm. Cirrus sac elongated, behind intestinal bifurcation, upto middle of ventral sucker, 0.31 - 0.46 mm x 0.07 - 0.09 mm. It includes an elongated vesicula seminalis, short pars prostatica, an ejaculatory duct and short cirrus. Ovary entire, transversely oval, pretesticular, 0.12 - 0.14 mm x 0.04 - 0.05 mm. An elongated saccular receptaculum seminis present just anterior to ovary. Uterus preovarian, short and opens at genital pore by a short metraterm. Genital pore just behind intestinal bifurcation. Vitellaria follicular, 0.02 - 0.03 mm x 0.01 mm x 0.02 mm, extending from hind end of ventral sucker or a little posterior to hind end of body. Follicles mainly lateral in position but behind posterior testis, intruding in intercaecal space. Excretory pore terminal. Excretory vesicle tubular, may or may not reach to posterior testis. Eggs few, oval, operculated, 0.11 - 0.16 mm x 0.15 - 0.22 mm.

Discussion

Looss (1900) erected the genus Allocreadium with A. isoporum as a type species. To the best of my knowledge, so far following species are described from India under the genus Allocreadium viz. A. handiai Pande, 1937; A. nicolli Pande, 1938 ; A. kosia Pande, 1938; A. schizothoracis Pande, 1938; A. mahaseri Pande, 1938; A. thapari Gupta, 1950; A. nemachilus Kaw, 1950; A. kamalai Gupta, 1956; A. mehrai Gupta, 1956; A. spindale Saksena, 1958; A. ophiocephali Srivastava, 1960; A. dollfusi Rai, 1962 ; A. singhi Rai, 1962 ; A. hirnai Rai, 1962; A. makundi Gupta, 1964; A. heteropneustusius Agrawal, 1964; A. catlai Kakaji, 1969; A. fasciatusi Kakaji, 1969; A. guptai Kakaji, 1969; A. calbasi Gupta & Puri, 1979; A. manteri Gupta & Puri, 1979; A. fotedari Dhar & Kharoo, 1984 and A. seenghali Sabhyata & Govind, 2008. The present form differs from A. handiai, A. nicolli, A. schizothoracis, A. mahaseri, A. thapari, A. nemachilus, A. dollfusi, A. singhi, A. hirnai, A. heteropneustusius, A. spindale, A. mehrai and A. kamalai in the extension of vitelline follicles. It also differ from A. kosia and A. mahaseri by the presence of short oesophagus and in the position of genital pore, from A. mehrai by the ratio of suckers and by the position of receptaculum seminis, from A. singhi by the ratio of suckers and absence of pre-pharynx, from A. macundi and A. heteropneustusius by the relative size of testes, from A. fasciatusi and A. guptai by the extension of vitelline follicles, absence of pre-pharynx. It further differs from A. fasciatusi by the ratio of suckers, position of receptaculum seminis and shape and position of cirrus sac, from A. guptai and A. catlai by the ratio of suckers, extension of cirrus sac and position of genital pore, shape and position of receptaculum seminis. Lastly, the present form differs from A. lucknowensis, A. indicum and A. fotedari in the extension of vitelline follicles, shape and extension of cirrus sac and position of genital pore. Therefore, the present form is regarded a new species and named as Allocreadium itwarensis n. sp. after the locality where from the host was procured.

0.2mm

Fig.4 Allocreadium itwaraensis n. sp. (Ventral view)

Allocreadium catlai Kakaji, 1969 (Fig. 5)

Host: Colisa fasciatus (Bloch & Schneider) Location: Body cavity Locality: Kanpur fish market No. of host examined: 10 No. of host found infected: 03 No. of worms/specimens collected: 08

Body (Fig.5) elongated, aspinose, 2.05 - 4.50 mm x 0.60 - 0.85 mm. Oral sucker terminal, ovoid, 0.22 - 0.39 mm x 0.23 - 0.40 mm. Ventral sucker round, pre-equatorial, more or less equal to oral sucker, 0.22 - 0.39 mm x 0.23 - 0.42 mm. Prepharynx absent. Pharynx muscular, ovoid, 0.10 - 0.20 mm x 0.12 - 0.21 mm. Oesophagus short, 0.08 - 0.10 mm x 0.16 - 0.20 mm. Caeca simple, form a short shoulder, extending upto posterior region of body. Testes entire, round to oval, tandem, postequatorial, anterior testis 0.20 - 0.52 mm x 0.31 - 0.50 mm; posterior testis large, close or slightly apart from anterior testis 0.31 - 0.51 mm x 0.31 - 0.56 mm. Cirrus sac anterior to ventral sucker, 0.20 - 0.26 mm x 0.07 - 0.09 mm. It contains a bipartite vesicula seminalis, pars prostatica, ejaculatory duct and a short cirrus. Ovary entire, oval, pre- equatorial, pretesticular, 0.12 - 0.29 mm x 0.12 - 0.25 mm. Receptaculum seminis elongated or pear-shaped, lateral to ovary, 0.21 - 0.27 mm x 0.06 - 0.12 mm. Uterus, pretesticular. Vitellaria follicular, lateral, extending from intestinal bifurcation upto hind end of body, but behind posterior testis, filling intercaecal space. Genital pore between intestinal bifurcation and ventral sucker. Eggs oval, few, non-operculated, 0.09 - 0.10 mm x 0.05 - 0.08 mm. Excretory bladder Y-shaped. Excretory pore terminal.

Discussion

A. catlai was discribed by kakaji (1969) from the intestine of Catla catla (Ham.) at Lucknow. This is another record of the worm from a new host and locality. The appended table shows the body measurements of worm as given by Kakaji (1969) and present form.

Table

Name of Allocreadium catlai Kakaji, 1969 Present specimens organs Body 3.36 mm x 1.06 mm 2.05 - 4.50 mm x 0.60 - 0.85 mm Oral sucker 0.27 mm 0.22 - 0.39 mm x 0.23 - 0.40 mm Pharynx 0.12 mm x 0.13 mm 0.10 - 0.20 mm x 0.12 - 0.21 mm Oesophagus 0.04 mm 0.08 - 0.10 mm x 0.16 - 0.20 mm V. Sucker 0.31 mm 0.22 - 0.39 mm x 0.23 - 0.42 mm Ovary 0.17 mm x 1.57 mm 0.12 - 0.29 mm x 0.12 - 0.25 mm Testes 0.30 - 0.34 mm x 0.28 - 0.30 mm 0.20 - 0.52 mm x 0.31 - 0.56 mm Cirrus sac 0.27 mm x 0.07 mm 0.20 - 0.26 mm x 0.07 - 0.09 mm Rec. seminis ? 0.21 - 0.27 mm x 0.06 - 0.12 mm Eggs 0.10 mm x 0.19 mm 0.09 - 0.10 mm x 0.05 - 0.08 mm

0.3mm

Fig.5 Allocreadium catlai Kakaji, 1969 (Dorsal view)

Allocreadium badatalabensis n. sp. (Fig. 6)

Host: Heteropneustes fossilis (Bloch) Location: Gut Locality: Bada Talab (Bhopal) No. of host examined: 06 No. of host found infected: 02 No. of worms/specimens collected: 12

Body (Fig.6) elongated, aspinose, broad anteriorly, narrow posteriorly 1.31 -1.65 mm x 0.42 - 0.62 mm. Oral sucker large, sub-terminal, 0.17 - 0.23 mm x 0.18 - 0.25 mm. Ventral sucker ovoid, smaller than oral sucker, 0.12 - 0.18 mm x 0.15 - 0.22 mm. Prepharynx absent. Pharynx muscular, ovoid, 0.06 - 0.10 mm x 0.06 - 0.10 mm. Oesophagus short, 0.05 -0.06 mm x 0.01 - 0.02 mm. Caeca simple extending upto hind end of body. Testes entire, oval, tandem, anterior testis 0.08 - 0.10 mm x 0.08 - 0.12 mm, posterior testis larger, 0.09 - 0.11 mm x 0.13 - 0.15 mm. Cirrus sac flask-shaped, median, between intestinal bifurcation and ventral sucker, 0.08 - 0.10 mm x 0.04 - 0.06 mm. Vesicula seminalis bipartite, pars prostatica tubular and surrounded by a large number of prostate gland cells. Ejaculatory duct short. Ovary entire, oval, pre-equatorial, pre-testicular, 0.07 - 0.10 mm x 0.13 - 0.19 mm. From anterior end of ovary arises oviduct which opens at ootype. Uterus short and opens at genital pore by a short metraterm. Genital pore located just behind intestinal bifurcation. Vitellaria follicular, extending from hind end of ventral sucker upto to posterior body extremity and occupy almost all intercaecal space behind ovary. Eggs large, oval, operculated, 0.04 - 0.07 mm x 0.06 - 0.08 mm. Excretory pore terminal, bladder tubular extending upto hind end of posterior testis.

Discussion

The present worm belongs to the genus Allocreadium Looss, 1900 for having pre-equatorial ventral sucker and less extensive vitellaria. Yamaguti (1971) listed 11 Indian valid species of the genus viz., A. handiai Pande, 1937; A. nicolli Pande, 1938; A. schizothoracis Pande, 1938; A. nemachilus Kaw, 1950; A. mehrai Gupta, 1956; A. dollfusi Rai, 1962; A. heteropneustusius Agrawal, 1964; A. makundi Gupta, 1964; A. catlai Kakaji, 1969 and A. fasciatusi Kakaji, 1969. Gupta and Verma (1976) described three new species Viz., A. mrighali, A. saranai and A. baranai. Later on Gupta and Puri (1979) added another two species A. calabasii and A. manteri of this genus. Sabhyata and Govind described A. seenghali from Varanasi. The new species differs from all the above species of the genus Allocreadium except A. handiai, A. Nicolli, A. kosia, A. thapari and A. heteropneustusius in having a larger oral sucker. It differs from A. nicolli and A. kosia in the presence of bipartite vesicula seminalis and position of genital pore between intestinal bifurcation and ventral sucker and from A. handiai and A. heteropneustusius in the commencement of vitellaria from the anterior level of ovary instead of from the posterior level of ventral sucker. It also differs from A. seenghali in position of genital pore and size of eggs. Accordingly, it is regarded as a new species with the specific name, Allocreadium badatalabensis n. sp. after the locality where from the host was procured.

100

0.3mm

Fig.6 Allocreadium badatalabensis n. sp. (Ventral view) 101

Eucreadium madhubanensis n. sp. (Fig.7)

Host: Eutropiicthys vacha (Ham.) Location: Gut Locality: Madhuban, District Mau No. of host examined: 08 No. of host found infected: 03 No. of worms/specimens collected: 08

Body (Fig.7) elongated, unarmed, 1.10 - 1.13 mm x 0.28 – 0.32 mm. Oral sucker 0.13 - 0.18 mm x 0.12 - 0.16 mm. Ventral sucker slightly larger than oral sucker, pre-equatorial, 0.12 - 0.18 mm x 0.11 - 0.17 mm. Pharynx muscular, 0.02 - 0.04 mm x 0.03 - 0.05 mm. Oesophagus tubular, slightly curved, 0.1 - 0.02 mm. Intestinal caeca extend upto hind region of body. Testes lobed, tandem, intercaecal, anterior testis, 0.12 - 0.13 mm x 0.15 - 0.17 mm; posterior testis 0.11 - 0.13 mm x 0.13 – 0.19 mm. Cirrus sac transversely elongated, 0.12 - 0.18 mm x 0.02 - 0.04 mm. Vesicula seminalis, transversely elongated, bipartite, pars prostatica, surrounded by prostate gland cells. Ejaculatory duct short and opens at genital pore by a short cirrus. Ovary entire, transversely elongated, pretesticular, 0.03 – 0.06 mm x 0.09 – 0.11 mm. Vitelline follicles mostly lateral, from oral sucker to hind end of body, in post testicular region, merge together. Uterine coils short and opens at genital pore by a short metraterm. Genital pore lateral, anterior to ventral sucker. posteriorly parallel to ovary. Eggs oval, operculated, 0.02 - 0.04 mm x 0.01- 0.03 mm. Excretory bladder tubular. Excretory pore at posterior end of body.

Discussion

Since the worm has small, elongated, unarmed body a large, subterminal oral sucker, short prepharynx, pharynx, caeca reaching to posterior 102

extremity, ventral sucker large, testes lobed, tandem, equatorial, cirrus sac pre-acetabular, genital pore sub-median, pre-acetabular, ovary lobed, between ventral sucker and anterior testis, vitellaria extending in lateral fields from pharynx to posterior extremity, confluent in post-testicular area, uterine coils confined to pre-testicular area, excretory vesicle tubular, therefore it belongs to the genus Eucreadium Dayal, 1942 with E. eutropiichthyius as type species. Subsequently, following species were added in India viz E. eucreadium Dayal, 1950; E. cameroni Gupta, 1963; E. jhingrani Srivastava & Singh, 1967; E. gangi Pandey, 1970; E. varanasi Agarwal & Verma, 1972; E. daceai Bashirullah and Mustaque, 1972; E. guptai Verma, 1973; E. thapari Agarwal & Kumar, 1979; E. hemlatae Gupta & Govind, 1983; E. pandeyi Srivastava, Saxena & Kumar, 1983 and E. kulpaharensis Agarwal and Agarwal, 1987. The present form chiefly differs from E. eutropiichthyius and E. eucreadium by the ratio of suckers and shape of ovary, from E. gangi by absence of pre-pharynx and entire testes, from E. jhingrani, E. daceai and E. kulpaharensis by the shape of testes, from E. cameronii, E. varanasi and E. thapari by the ratio of suckers and extension of vitellaria and from E. guptai and E. hemlatae by the position of gonads. It further differs from E. hemlatae by the extension of vitellaria. Therefore, the present worm is described as a new species and named as Eucreadium madhubanensis n. sp.

103

0.2mm

Fig.7 Eucreadium madhubanensis n. sp. (Ventral view)

104

Rhynchocreadium dohrighatensis n. sp. (Fig. 8)

Host: Channa marulius (Ham.) Location: Body cavity/ Gut Locality: Dohrighat No. of host examined: 10 No. of host found infected: 03 No. of worms collected: 08

Body (fig.8) small, elongated, cylindrical, smooth, crescent shaped, 1.33 - 2.85 mm x 0.41 - 0.88 mm. Oral sucker subterminal, round, 0.01 - 0.02 mm x 0.01 mm x 0.02 mm. Ventral sucker large, pedicellate, 0.12 - 0.23 mm x 0.15 - 0.30 mm. Prepharynx absent. Pharynx muscular, small, well marked, oval, 0.07 - 0.09 mm x 0.08 - 0.09 mm. Oesophagus long, slender, 0.15 mm x 0.27 mm, bifurcates into two intestinal caeca, slightly anterior to ventral sucker. Caeca almost upto posterior extremity, masked by vitellaria. Testes tandem, almost equal; anterior testis 0.14 - 0.20 mm x 0.13 - 0.15 mm; posterior testis 0.14 - 0.22 mm x 0.13 - 0.15 mm. Vasa efferentia from both testes run anteriorly, meet near ventral sucker, before opening into cirrus sac. Cirrus sac 0.16 - 0.21 mm x 0.07 - 0.08 mm, club-shaped, pre acetabular. Vesicula seminalis bipartite. Cirrus muscular. Ovary round to oval, close to ventral sucker, pre-testicular, pre-equatorial, 0.10 - 0.22 mm x 0.06 - 0.19 mm. Genital pore lateral, just anterior to ventral sucker. Vitelline glands extend from posterior margin of ovary to hind region of body, filling intercaecal space. Uterus pre testicular, with few eggs. Eggs small, non filamentous, operculated, 0.02 - 0.06 mm x 0.008 - 0.01 mm. Excretory pore at posterior end of body, terminal excretory bladder long, tubular.

Discussion

Since the worm has small cylindrical body, terminal oral sucker, pedicellate ventral sucker, prepharynx absent, pharynx small, oesophagus 105

slender, caeca almost at posterior extremity, testes oval, tandem, vesicula seminalis bipartite, cirrus pouch preacetabular, ovary round, pretesticular, vitellaria from ovarian level to posterior end of body, genital pore lateral, preacetabular, excretory vesicle reaching uoto ventral sucker, therefore, it belongs to the genus Rhynchocreadium Srivastava, 1962. To the best of my knowledge, only the type species R. aculeatum Srivastava, 1962 is so far known under the genus. The present worm chiefly differs from above species in ratio of suckers, ratio of testes and ovary and extension of vitellaria. Therefore, it is regarded as a new species and named R. dohrighatensis n. sp. where from the host was procured.

106

0.2mm

Fig.8 Rhynchocreadium dohrighatensis n. sp. (Ventral view)

107

Neopodocotyle dohrighatensis n. sp. (Fig. 9)

Host: Channa puctatus (Bloch.) Location: Gut Locality: Dohrighat No. of host examined: 15 No. of host found infected: 03 No. of worms collected: 06

Body (Fig.9) elongated, aspinose, rounded at extremities, 2.08 - 2.53 mm x 0.51 - 0.69 mm. Oral sucker subterminal, 0.26 - 0.30 mm x 0.24 - 0.31 mm. Ventral sucker smaller than oral sucker, pre-equatorial, 0.20 - 0.22 mm. Prepharynx absent. Pharynx muscular, ovoid, 0.08 - 0.11 mm x 0.10 - 0.11 mm. Oesophagus short, tubular, 0.03 - 0.11 mm. Intestinal caeca simple, extending to posterior testis. Genital pore median or on left side of body, intercaecal, either close or apart from intestinal bifurcation, 0.56 - 0.66 mm from anterior extremity. Excretory pore lies at posterior end of body. Excretory bladder tubular, extending upto hind end of posterior testis. Testes entire, oval, tandem, intercaecal, anterior testis smaller than posterior testis, 0.19 - 0.27 mm x 0.15 - 0.22 mm and posterior testis 0.28 - 0.45 mm x 0.19 - 0.23 mm. Cirrus sac flask-shaped, 0.12 - 0.22 mm x 0.08-0.10 mm, lies below intestinal bifurcation and ventral sucker. Vesicula seminalis bipartite, proximal part 0.03 - 0.06 mm x 0.03 - 0.50 mm; distal part 0.02 -0.05 mm x 0.02 - 0.04 mm. Pars prostatica opens into a muscular cirrus through an ejaculatory duct, 0.04 - 0.07 mm. Space around vesicula seminalis and pars prostatica surrounded by a large number of prostate gland cells. Ovary entire, oval, equatorial, pre-testicular, lying between ventral sucker and anterior testis, 0.18 - 0.21 mm x 0.13 - 0.17 mm. From anterior end of ovary arises oviduct which opens at ootype. Receptaculum seminis elongated, 0.12-0.20 mm x 0.05-0.10 mm, lying laterally on left or right side of ovary or a little 108

anterior to it, overlapping hind end of ventral sucker. Vitellaria follicular, extending from posterior region of ventral sucker or a little posterior to it upto a little infront of hind end of body. Follicles mainly lateral in position but behind posterior testis, intruding in intercaecal space. Eggs large, oval, operculated, few, 0.07 - 0.08 mm x 0.04 - 0.05 mm.

Discussion

To the best of my knowledge besides the type species N. indica Dayal, 1950 following more species are known under the genus viz. N. lucknowensis Gupta & Chakrabarti, 1966; N. mehrai Rai, 1971; N. spinipora Sircar and Sinha, 1969, N. dayali Pandey, 1973 and N. balliaensis Pandey 1973. The present worm chiefly difers from all the above species in ratio of sucker, position of genital pore and size of egg. Therefore, it is regarded a new species and named N. doharighathensis n. sp. after the locality where from the host was procured.

109

0.2mm

Fig.9 Neopodocotyle dohrighatensis n. sp. (Ventral view)

110

Peracreadium bhopalensis n. sp. (Fig. 10)

Host: Macrognathus aculeatus (Bloch) Location: Gut Locality: Bada Tal (Bhopal) No. of host examined: 07 No. of host found infected: 03 No. of worms collected: 08

Body (Fig.10) elongated, small, aspinose, 1.8 - 0.20 mm x 0.68 - 0.72 mm. Oral sucker subterminal, more or less circular, 0.13 - 0.15 mm x 0.14 - 0.16 mm. Prepharynx absent. Pharynx muscular, oval or sub-spherical, 0.07 - 0.08 mm x 0.08 - 0.10 mm. Oesophagus short, 0.06 - 0.09 mm x 0.01 – 0.02 mm. Intestinal caeca extending upto hind region of body. Ventral sucker large, spherical, pre-equatorial, 0.28 - 0.32 mm x 0.27 - 0.30 mm. Testes transversely elongated, tandem, postequatorial, anterior testis 0.15 – 0.18 mm x 0.25 - 0.30 mm, and posterior testis 0.16 - 0.20 mm x 0.30 - 0.35 mm. Cirrus sac large, crescent-shaped, lying on left side of body, 0.65 - 0.68 mm x 0.09 - 0.11 mm, extending behind ventral sucker. It includes an elongated vesicula seminalis, a prominent pars-prostatica, surrounded by prostrate gland, a short ejaculatory duct and a cirrus. Cirrus muscular, non-spiny. Ovary large, pre- testicular, transversely elongated, 0.19 - 0.21 mm x 0.32 - 0.33 mm. A receptaculum seminis present. Vitellaria follicular, small, extending from hind end of body upto anterior border of ventral sucker, 0.03 - 0.07 mm x 0.04 - 0.08 mm. Uterus short, pre-testicular, and opens at genital pore by short metraterm. Excretory bladder tubular and opens outside through an internal excretory pore. Genital pore behind intestinal bifurcation. Eggs oval, non- operculated, few in number, 0.03 - 0.04 mm x 0.01 - 0.02 mm.

111

Discussion

Since the worm has an elongated body, oral sucker and pharynx well developed, oesophagus short, caeca terminating at posterior extremity, Ventral sucker larger than oral sucker, pre-eqatorial. Testes tandem, post-eqatorial, cirrus sac long, extending posterior to ventral sucker, enclosing, seminal vesicle, genital pore median, near intestinal bifurcation, ovary sub median, immediately pre-testicular, receptaculum seminis present, vitelline follicles, extending in fore-and hind body, uterus comperatively short, winding between ovary and testes and genital pore, excretory vesicle short, therefore it belongs to the genus Peracreadium Nicoll, 1909. To the best of my knowledge so for following species of the worm Peracreadium are known viz. P. genu (Rudolphi, 1819) Nicoll, 1909; P. perezi Mathias, 1926; P. shawi (Mentosh, 1939) Winter, 1959; P. mycteropercae (Sogandares Bernal, 1959) Pritchard, 1966 and P. guptai Kakaji, 1969. Thus, the genus has only one species viz. P. guptai described by Kakaji (1969) in India. The present worm chiefly differs from all the above species in ratio of sucker, shape and size of gonads and size of eggs. Therefore, it is regarded a new species and named Peracreadium bhopalensis n. sp. after the locality wherefrom the host was procured.

112

0.2mm

Fig.10 Peracreadium bhopalensis n. sp. (Ventral view)

113

Genarchopsis goppo Ozaki, 1925 (Fig. 11)

Host: Wallago attu (Bl. & Schn.) Location: Gut Locality: Itwara fish market (Bhopal) No. of host examined: 03 No. of host found infected: 01 No. of parasites collected: 02

Body (Fig.11) elongated or spindle like, 2.65 - 3.02 mm x 0.85 - 1.1 mm. Oral sucker subterminal, oval, 0.25 - 0.35 mm x 0.38 - 0.44 mm. Prepharynx absent. Pharynx muscular, well developed, 0.10 - 0.14 mm x 0.08 - 0.15 mm. Oesophagus short, 0.02 - 0.04 mm x 0.01 - 0.03 mm. Intestinal caeca forming a shoulder, with crenated margin extend upto hind region of body and unite at posterior extremity. Ventral sucker larger than oral sucker, post-equatorial, 0.60 - 0.64 mm x 0.62 - 0.66 mm. Testes oval, post-acetabular, subequal, parallel, 0.20 - 0.24 mm x 0.14 - 0.19 mm. Cirrus sac absent. Vesicula seminalis elongated, saccular, 0.30 - 0.39 mm x 0.12 - 0.18 mm. Pars prostatica surrounded by a large number of prostate gland cells. Ejaculatory duct short. Ovary oval, submedian, near posterior extremity, 0.13 - 0.18 mm x 0.18 - 0.23 mm, post acetabular, post testicular. Receptaculum seminalis absent. Vitellaria consist of two, lobed, bean shaped masses, lying in posterior region of body. Right vitelline gland 0.37 - 0.41 mm x 0.26 - 0.29 mm and left 0.30 - 0.35 mm x 0.20 – 0.25 mm. Vitelline ducts arise from each lobe and join each other, before opening at ootype. Uterus arises from ootype, forms closely packed transverse coils, occupying all pre and post acetabular space. Genital pore near intestinal bifurcation. Excretory pore at hind end of body. Bladder Y-shaped. Eggs oval, non-operculated, 0.03 - 0.05 mm x 0.01- 0.02 mm.

114

Discussion

Ozaki (1925) established the genus Genarchopsis with G. goppo as the type species. For Genarchopsis mulleri Levinson, 1881 Looss (1899) created the genus Progonus. However, Srivastava (1933) regarded Genarchopsis as synonym of Progonus and transferred P. piscicola and P. ovocaudatum to the genus and established Ophiochorchis to include O. lobatum Srivastava (1933) and O. singularis Srivastava (1933) due to the presence of an oesophageal pouch. Gupta (1951) agreed to Srivastava (1933) and added O. dasus, O. indicus and O. faruquis to the genus Ophiochorchis. However, Chauhan (1953) doubted the validity of certain species of the genus Genarchopsis and Ophiochorchis and pointed out the wrong nomenclature given by Gupta (1951) for O. dasus and O. faruquis. Skrjabin et al., (1955) retained the genus Ophiochorchis and its five species valid because of the presence of oesophageal pouch. Yamagutti (1958) considered the genus Genarchopsis valid, considered with Progonus and Ophiochorchis its synonym and listed 11 species under the genus Genarchopsis of which eight were transferred from the genera Ophiochorchis and Progonus. The writer fully agrees with Yamagutti (1958) and regards Ophiochorchis and Progonus as synonym of Genarchopsis. Till now, the genus Genarchopsis includes 18 species viz. G. goppo (Tubangui) Ozaki, 1925 type species; G. gigi Yamaguti, 1919; G. anguillae Yamaguti, 1938; G. mulleri (Levinson, 1818); G. lobatum (Srivastava, 1933); G. ovocaudatum (Srivastava,1933); G. piscicola (Srivastava,1933); G. singularis (Srivastava,1933) G. dasus (Gupta,1951); G. faruquis (Gupta,1951); G. indicus (Gupta,1951); G. melanostictus Dwivedi, 1965; G. thapari Gupta and Chakrabarti, 1966; G. punctati Agrawal, 1966; G. cuchiai Kakaji, 1969; G. cameroni Kakaji, 1969; G. ozakii Bashirullah and Elahi, 1972; G. Bangladesensis Bashirullah and Elahi, 1972. Of these, Pandey (1973) considered only type species G. goppo valid and others as its synonyms, to which I also agree. 115

0.2mm

Fig.11 Genarchopsis goppo Ozaki, 1925 (Ventral view)

116

Haplorchoides varanasiensis n. sp. (Fig. 12)

Host: Eutropicthes vacha (Ham.) Location: Gut Locality: Fish market, Varanasi No. of host examined: 15 No. of host found infected: 08 No. of worms/specimens collected: 22

Body (Fig.12) small, pear shaped, broad anteriorly and narrow posteriorly, 1.02 - 1.27 mm x 0.51 - 0.63 mm, covered with small spines. Oral sucker terminal, oval, 0.08 - 10 mm x 0.10 - 0.11 mm. Prepharynx, 0.10 mm x 0.02 mm. Pharynx 0.03 - 0.04 mm x 0.04 - 0.05 mm. Oesophagus short, longer than prepharynx, 0.05 mm x 0.02 mm, leading into intestinal caeca, which run close to body wall, terminate in posterior half of post-testicular region. Ventral sucker absent. Testis large, transversely elongated, with irregular margin, post equatorial, 0.10 - 0.13 mm x 0.22 - 0.26 mm. Cirrus sac absent. Seminal vesicle joint with metraterm through small ejaculatory duct and opens into a ventrogenital complex (Gonotyle), behind intestinal bifurcation. Ventrogenital sinus, contain embedded ventral sucker, with circlet of about 16-24 weakely chitinised spines, arranged in a single row. Ovary entire, oval, pre-equatorial lying between ventro-genital sac and testis, 0.07-0.09 mm x 0.06-0.09 mm. Receptaculum seminalis oval, larger than ovary, anterior side to ovary, 0.10 - 0.12 mm x 0.11 - 0.15 mm. Genital sac opens through a groove on gonotyl. Gonotyl intercaecal, lying on left side of body, just behind intestinal bifurcation, 0.03 - 0.04 mm. Uterus fills entire space from intestinal bifurcation to hind region of body. Vitelline glands of large follicles, mainly on lateral sides, sometimes covering intestinal ceaca, irregular in shape, extending from posterior end of receptaculum seminis upto hinder end of intestinal caeca. Excretory bladder Y-shaped, elongated, opens at posterior end of body by a terminal aperture. Eggs oval, operculated, yellowish brown, 0.03 - 0.04 mm x 0.01 - 0.02 mm. 117

Discussion

The genus Haplorchoides Chen, 1949 includes following species viz. H. chirinus Looss, 1896; H. attenuatum Srivastava, 1935; H. piscicola Srivastava, 1935; H. taakree (Dayal, 1935); H. macrones (Dayal, 1949); H. gomtioensis Gupta, 1953; H. ritai Gupta, 1953; H. brahamputraensis Gupta, 1953; H. seenghali Gupta, 1953; H. parini Chatterjii, 1956; H. macronis Agrawal, 1964; H. pearsoni Pande and Shukla, 1976; H. mehrai Pande and Shukla, 1976; H. kherai Gupta & Govind, 1985; H. srivastavi Gupta & Govind, 1985; H. rajghatai Maurya & Agarwal, 1999; H. hardayali Maurya & Agarwal, 1999; H. mystusi Maurya & Agarwal, 1999 and H. dayali Maurya & Agarwal, 1999. The present worm chiefly differs from all above species in the position of ventrogenital sac, number of spines in ventrogenital sac and extension of vitellaria. It is therefore, considered a new species H. varanasiensis n. sp named after the locality wherefrom the host was procured.

118

0.2mm

Fig.12 Haplorchoides varanasiensis n. sp. (Ventral view)

119

Opisthorchis pedicellata Verma, 1927 (Fig. 13)

Host: Rita rita (Ham.) Location: Gall bladder Locality: Fish market, Lucknow No. of host examined: 05 No. of host found infected: 02 No. of worms collected: 03

Body (Fig.13) elongated, flattened, spinose, 4.25 - 8.70 mm x 0.13 - 0.95 mm. Oral sucker oval, terminal, 0.12 - 0.14 mm x 0.18 - 0.20 mm. Ventral sucker almost equal or slightly larger than oral sucker, 0.13 - 0.15 mm x 0.19 - 0.21 mm. Prepharynx present. Pharynx 0.06 - 0.08 mm x 0.07 - 0.09 mm. Oesophagus prominent 0.04 mm x 0.01 mm. Intestinal caeca extend upto hind end. Testes tandem, more or less lobed, close to each other in posterior region of body, 0.30 - 0.46 mm x 0.15 - 0.30 mm. Genital pore median, preacetabular. Ovary rounded, pretesticular, 0.20 - 0.26 mm x 0.09 - 0.12 mm. Receptaculum seminis prominent, oval, oblique, between ovary and anterior testis. Uterus extends from ovary to ventral sucker, touching caeca, sometimes overlapping and consists of convolutions. Eggs oval, 0.01 - 0.02 mm x 0.01 - 0.01 mm. Vitellaria from a little behind the ventral sucker to ovarian region, 0.02 - 0.04 mm x 0.02 - 0.03 mm.

Discussion

Verma (1927) described the worm from gall bladder of Rita rita at Allahabad. This is another record of the worm from Lucknow. The appended table shows the body measurements of worm, given by Verma, 1927 and present species.

120

Table

Name of Opisthorchis pedicellata Verma, Present specimens organs 1927 Body 6.3 - 12.7 mm x 0.23 - 1.6 mm 4.25 - 8.70 mm x 0.13 - 0.95 mm. Oral sucker 0.20 mm x 0.30 mm 0.12 - 0.14 mm x 0.18 - 0.20 mm Pharynx 0.13 mm x 0.19 mm 0.06 - 0.08 mm x 0.07 - 0.09 mm Oesophagus ? 0.04 mm x 0.01 mm V. Sucker 0.28 mm x 0.39 mm 0.13 - 0.15 mm x 0.19 - 0.21 mm Ovary ? 0.20 - 0.26 mm x 0.09 - 0.12 mm Testes 0.40 - 0.56 mm x 0.19 - 0.35 mm 0.30 - 0.46 mm x 0.15 - 0.30 mm Eggs 0.03 mm x 0.016 mm 0.01 - 0.02 mm x 0.01 - 0.01 mm

121

0.2mm

Fig.13 Opisthorchis pedicellata Verma, 1927 (Ventral view)

122

Oudhia itwaraensis n. sp. (Fig. 14)

Host: Heteropneustes fossilis (Bloch) Location: Gut Locality: Itwara fish market (Bhopal) No. of host examined: 10 No. of host found infected: 03 No. of worms collected: 12

Body (Fig.14) elongated, aspinose, rounded at extremities, 2.08 - 2.53 mm x 0.51 - 0.69 mm. Oral sucker terminal, oval, 0.26 - 0.30 mm x 0.24 - 0.31 mm. Ventral sucker equal to oral sucker, pre-equatorial, 0.22 - 0.28 mm x 0.21 x 0.27 mm. Prepharynx present, 0.05 mm x 0.01 mm. Pharynx muscular, ovoid, 0.08 - 0.11 mm x 0.10 - 0.11 mm. Oesophagus short, tubular, 0.03 - 0.11 mm, bifurcates into simple intestinal caeca, extending upto hind region of body. Intestinal caeca simple, reaching upto hind region of body. Testes entire, oval, located nearly in mid region of body, obliquely tandem, intercaecal, partly overlapping each other. Anterior testis 0.19 - 0.27 mm x 0.15 - 0.25 mm and posterior testis 0.28 - 0.45 mm x 0.19 - 0.23 mm. Cirrus sac long, tubular, consists of an elongated saccular distal part and a narrow tubular proximal part, 0.12 - 0.22 mm x 0.08 - 0.10 mm, lies on right side of body, extending from genital pore to anterior end of ventral sucker. It includes a bulbous vesicula seminalis. Pars prostatica is surrounded by a large number of prostate gland cells and is continued forward into a long narrow, tubular ejaculatory duct which ultimately terminates into small unarmed cirrus. Ovary entire, oval, pre-equatorial, pre-testicular, lying diagonal to ventral sucker, larger than testes, 0.358 - 0.52 mm x 0.22 - 0.26 mm. Receptaculum seminis oval, 0.03 - 0.08 mm x 0.04 - 0.08 mm, just behind ovary. Uterus occupies almost all space in hind region of the body and opens anteriorly at genital pore by a short metraterm. Vitellaria follicular, extending from ventral sucker or a 123

little posterior to it upto testes. Follicles mainly lateral in position, some lie posterior to ventral sucker as well. Excretory bladder tubular, extending upto hind region of posterior testis. Genital pore is located behind posterior border of oral sucker. Eggs numerous, operculated, oval, 0.02 - 0.03 mm x 0.01 - 0.02 mm.

Discussion

The worm has spinose body, oral sucker funnel-shaped, prepharynx present, pharynx small, oesophagus very short, caeca terminating at hind end of body, ventral sucker in anterior half of body, testes obliquely overlapping each other behind ovary, cirrus pouch long, claviform enclosing bipartite seminal vesicle, well developed prostatic complex and long ejaculatory duct, genital pore sinistral to prepharynx, ovary subterminal, pretesticular, postacetabular, vitelline glands extending from middle of ventral sucker to posterior region of anterior testis, excretory bladder Y- shaped, eggs oval, operculated, therefore, it belong to the genus Oudhia Gupta, 1955. To the best of my knowledge apart form O. herai the type species, O. hardayali has been added by Kumar and Agarwal (1980) from the intestine of M. vittatus at Varanasi. The present species differs from both the above species, in ratio of suckers, extention of vitellaria and size and ratio of overy and testes. Furthermore, the position of genital pore further differentiates the species from described species. Therefore, it is regarded a new species and named O. itwarensis, n. sp. after the locality where from the worm was collected.

124

0.2mm

Fig.14 Oudhia itwaraensis n. sp. (Ventral view)

125

Phyllodistomum vachius Dayal, 1949 (Fig. 15)

Host: Mastacembalus puncalus (Ham.) Location: Urinary bladder Locality: Bada Talab (Bhopal) No. of host examined: 07 No. of host found infected: 03 No. of metacercariae collected: 14

Body (Fig.15) spatulate, divided into fore and hind body, 1.60 - 1.80 mm x 0.65 - 0.82 mm. Fore body tapering anteriorly, hind body foliate. Oral sucker terminal, 0.15 - 0.23 mm x 0.20 - 0.22 mm. Ventral sucker in middle of body, larger, 0.25 - 0.30 mm x 0.26 - 0.31mm. Pharynx absent. Oesophagus long, slender, 0.20 - 0.22 mm x 0.14 - 0.20 mm. Intestinal Caeca simple, reaching upto hind region of body. Testes lobed, intercaecal, diagonal, anterior testis 0.20 - 0.23 mm x 0.19 - 0.23 mm and posterior testis 0.15 - 0.20 mm x 0.12 - 0.19 mm. Vesicula seminalis saccular, bipartite, anterior chamber, 0.06 - 0.10 mm x 0.04 - 0.06 mm; posterior chamber 0.04 - 0.07 mm x 0.03 - 0.05 mm. Ovary oval, submedian, pre-testicular, 0.10 - 0.13 mm x 0.07 - 0.10 mm. Uterus runs in hind body, occupying post acetabular space and opens at genital pore by a short metraterm. Genital pore post-bifurcal. Excretory vesicle long, tubular and lies between testes. Eggs few, oval, thin, 0.02 - 0.04 mm x 0.01 - 0.02 mm.

Discussion

Dayal (1949) described Phyllodistomum vachius from the urinary bladder of Eutropiicthys vacha at Lucknow. It was also recorded by Pandey (1973) from the urinary bladder of H. fossilis at Lucknow. This is another record of the worm from a new host Xenentodon cancila and locality. The present worms differ from those described by Dayal (1949) in having straight 126

oesophagus, more or less equal testes and tubular excretory bladder. The looped oesophagus described by Dayal (1949) appears to be due to contraction in the anterior part of the body. The appended table shows the body measurements of P. vachius as given by Dayal, (1949) and present specimens.

Table

Name of Phyllodistomum vachius Dayal, Present specimens organs 1949 Body 7.1 mm x 5.0 mm 1.60 - 1.80 mm x 0.65 - 0.82 mm Oral sucker 0.75 mm x 0.60 mm 0.15 - 0.23 mm x 0.20 - 0.22 mm Oesophagus ? 0.20 - 0.22 mm x 0.14 - 0.20 mm V. Sucker 0.50 mm 0.25 - 0.30 mm x 0.26 - 0.31mm Ovary 0.35 mm x 0.45 mm 0.10 - 0.13 mm x 0.07 - 0.10 mm Testes 0.35 - 0.45 mm x 0.40 - 0.45 mm 0.12 - 0.23 mm x 0.15 - 0.24 mm Cirrus sac 0.50 mm x 0.30 mm 0.04 - 0.10 mm x 0.03 - 0.06 mm Eggs 0.035-0.038 mm x 0.027-0.029 mm 0.02 - 0.04 mm x 0.01 - 0.02 mm

I would like to further supplement that following species are known from India viz. P. lewisi Srivastava, 1938; P. macronius (Dayal, 1938) Yamaguti, 1958 Syn: Phyllochorus macronius Dayal, 1938, P. callichrous (Dayal, 1949) Yamaguti, 1958 Syn: Plesiodistomum callichrous Dayal, 1949; P. vachius Dayal, 1949; P. loossi Kaw, 1950; P. singhiai Gupta, 1951; P. vittatusi Gupta, 1953; P. chauhani Motwani & Srivastava, 1961; P. tripathii Motwani & Srivastava, 1961; P. srivastavi Rai, 1964; P. cameroni Agarwal, 1966; P. parorchium Jaiswal, 1957; P. indianum Jaiswal, 1957; Phyllodistomum sp. Jaiswal, 1957; P. lucknowensis Pandey, 1971; P. longicephalus Singh & Sinha, 1975; P. bimaculatusi Ahmad, Singh & Maurya, 1999 and P. guptai Shomorendra & Jha, 2006. Out of these, Rai (1971) has considered the following species valid viz., P. lewisi Srivastava 1938 (P. sp. Bhalerao); P. tripathi Motwani and 127

Srivastava, 1961; P. loosi Kaw, 1950 (P. camaroni Agarwal, 1966); P. srivastavai Rai, 1964; P. lucknowensis Pandey, 1970. On the persual of literature I am convinced that P. tripathii, P. loosi, P. parachis, P. indicum, P. srivastavai, P. lucknowensis P. longicephalus closely resemble with P. lewisi and not valid in view of the variable characters like size of body, position of gonads and shape of seminal vesicle. On which they are based. They are, therefore, further considered synonyms of P. lewisi. The other Indian species P. bimaculatusi closely resembles with P. vachius and therefore considered as synonym of P. vachius. Shomorendra and Jha (2006) have reported a pharynx in P. guptai. I doubt the observation, as a pharynx is absent in the genus Phyllodistomum. However, if it is present, P. guptai must be removed from the genus Phyllodistomum under a separate genus. I would also like to pointout that except for presence of pharynx, P. guptai simulates P. vachius in almost all the body features.

128

0.3mm

Fig.15 Phyllodistomum vachius Dayal, 1949 (Ventral view)

129

Pleurogenoid madhubanensis n. sp. (Fig.16)

Host: Ompok bimaculatus (Bloch) Location: Gut Locality: Madhuban Tal No. of host examined: 15 No. of host found infected: 04 No. of worms collected: 10

Body (Fig.16) elliptical, spinose, posterior extremity bifid, 0.70 - 0.88 mm x 0.34 - 0.46 mm. Oral sucker terminal, oval, 0.09 - 0.11 mm x 0.10 - 0.19 mm. Ventral sucker equatorial, equal to oral sucker, 0.10 - 0.13 mm x 0.08 - 0.12 mm. Pre-pharynx very small. Pharynx bean shaped, muscular, 0.02 - 0.03 mm x 0.04 - 0.06 mm. Oesophagus long, 0.06 - 0.10 mm x 0.01 - 0.02 mm, bifurcating into two intestinal caeca, extending upto testes. Testes elongate-oval, entire, equatorial, laterally placed, parallel, right testis 0.11 - 0.15 mm x 0.06 - 0.08 mm, and left testis 0.11 - 0.16 mm x 0.05 - 0.07 mm. Cirrus sac elongate-oval, on side of oesophagus upto ventral sucker. It contains a large vesicula seminalis, pars prostatica, surrounded by prostrate gland and short ejaculatory duct opening at genital pore. Ovary oval, pre-testicular, 0.08 - 0.14 mm x 0.05 - 0.09 mm. Oviduct arises from middle of its inner margin, opens at ootype. Vitellaria follicular, consisting of 12 to 17 follicles extending from level of pharynx to ovary, 0.04 - 0.06 mm x 0.01 - 0.03 mm. Two vitelline ducts unite to form a common duct to open at ootype. Uterus coiled, with descending and ascending limbs and opens at genital pore by a short metraterm. Genital pore lateral, at level of pharynx. Eggs oval, non-operculated, 0.01-0.029 mm x 0.011- 0.019 mm. Excretory bladder Y-shaped and opens by terminal excretory pore.

130

Discussion

The genus Pleurogenoides Travassos, 1921 includes following species P. tener (Looss, 1896) Travassos, 1921; P. medians (Olsson, 1876) Travassesos, 1921; P. arrectus (Duj, 1845) Dollfus, 1961. P. minus (Pigulewsky, 1931) Kaw, 1943; P. pabdai Pande, 1937; P. japonicus Yamaguti, 1936; P. sitapurii (Srivastava, 1934), Kaw, 1948; P. notopteris Bashirullah and Hafijuddin, 1976 and P.attui (Kakaji, 1968) Yamaguti, 1971. The present species differs from P. japanicus, P. sitapuri, P. minus and P. notopteris by having a bifid posterior body and extension of vitellaria. It further differs from P. attui and P. pabdai by the presence of prepharynx and shape of excretory bladder. It differs from P. minor by the position of gonads and genital pore and from P. attui by the ratio of suckers and extension of cirrus. Therefore, the present form is regarded as a new species and named as madhubanensis n. sp. The species has been named after locality wherefrom the host was procured.

131

0.2mm

Fig.16 Pleurogenoid madhubanensis n. sp. (Ventral view)

132

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SUMMARY

The work incorporated in the thesis entitled "ON SOME METACERCARIAE AND ADULT TREMATODES OF FRESHWATER FISHES" deals with the taxonomical studies of trematode parasites of fresh water fishes. It includes description of 22 metacercariae and 16 adult trematodes, found infecting freshwater fishes. Fishes for the present work were collected from fish markets and water bodies of districts Aazamgarh, Allahabad, Ballia, Basti, Belthara, Bhagalpur, Bhatpar Rani, Bhopal (M. P.), Bijnor, Chapra (Bihar), Deoria, Dohrighath, Faizabad, Gonda, Gorakhpur, Jhansi, Kanpur, Lakhimpur Khieri, Lucknow, Madhuban, Mau, Meerut, Pallia, Sahi (U. P.) and Varanasi during 2007 - 2011. Standard methods of killing, fixing, staining and mounting of adult trematodes and metacercariae were used. Camera lucida diagrams were made. The work is divided into 2 parts viz. Part-I and Part-II.

Part-I

Part -Ideals with the description of 22 metacercariae.

1. Neascus bhopalensis n. sp. was collected from the liver of Xenentodon cancila (Ham), bought from Itwara fish market, Bhopal. The new species is characterized by ratio of fore and hind body, absence of pharynx, ratio of suckers, position of holdfast organ, shape of holdfast gland and number of genital rudiments.

2. Neascus dohrighatensis n. sp. was collected from liver of Wallago attu (Bl. & Schn.), at Dohrighath. The new species is characterized by ratio of fore and hind body, absence of pharynx and shape of holdfast organ and holdfast gland.

3. Neascus khurramnagarensis n. sp. was collected from the liver of Xenentodon cancila (Ham.), bought from Khurram Nagar fish market, 171

Lucknow. The new species is characterized by shape of body, ratio of fore and hind body, shape of holdfast organ and holdfast gland and number of genital rudiments.

4. Neascus kaisarbaghensis n. sp. was collected from cranium/branchial region of Anabas testudineus (Bl.), bought from Kaisarbagh fish market, Lucknow. The new species is characterized by ratio of fore and hind body, absence of pharynx, intestinal caeca reaching upto posterior end of hind body, ratio of suckers and shape of holdfast organ and holdfast gland.

5. Tetracotyle bhopalensis n. sp. was collected from gut of Channa punctatus (Bloch), bought from fish market, Bhopal. The new species is characterized by pre-equatorial ventral sucker, larger than oral sucker, and number of genital rudimentss.

6. Tetracotyle mauensis n. sp. was collected from liver/gut of Xenentodon cancila (Ham.), at Mau (U.P.). The new species is characterized by oval, thick, two layered cyst, shape and size of pseudosuckers and number of genital rudiment.

7. Tetracotyle allahabadensis n. sp. was collected from gut of Mastacembelus puncalus (Ham.), bought from fish market, Allahabad. The new species is characterized by absence of pre-pharynx, short oesophagus, two genital rudiments and bowl-shaped hold fast organ.

8. Tetracotyle madhubanensis n. sp. was collected from branchial region of Mastacembelus puncalus (Ham.), bought from fish market, Madhuban. The new species is characterized by large lateral pseudosuckers, presence of pre-pharynx, extension of intestinal caeca upto holdfast organ and number of genital rudiments.

9. Tetracotyle saiensis n. sp. was collected from the body cavity of Colisa fasciatus (Bl. & Schn.), caught from river Sai, Lucknow. The new species 172

is characterized by ratio of suckers, shape and position of pseudosuckers and shape of holdfast organ and holdfast gland.

10. Tetracotyle daliganjensis n. sp. was collected from liver of Xenentodon cancila (Ham.), bought from Daliganj fish market, Lucknow. The new species is characterized by shape and position of pseudosuckers, by shape and position of holdfast organ and number of genital rudiment.

11. Tetracotyle megapseudosuckerai n. sp. was collected from liver of fish Mastacembelus armatus (Lacep.), caught from river Gomti, Daliganj. The new species is characterized by shape of body, ratio of suckers and position of holdfast organ and holdfast gland.

12. Tetracotyle multilobulata n. sp. was collected from cranium of Macrognethus aculeatus (Bloch) caught from river Gomti, Daliganj. The new species is characterized by long oesophagus, intestinal caeca reaching upto holdfast organ, hold fast organ situated behind ventral sucker and number of genital rudiments.

13. Tetracotyle varanasiensis n. sp. was collected from gut of fish Macrognethus aculeatus (Bloch), bought from fish market, Varanasi. The new species is characterized by a collar at anterior region of body, ratio of suckers, position of holdfast organ and holdfast gland and number of genital rudiments.

14. Tetracotyle trilobulata n. sp. was found in liver of Mastacembelus armatus (Lacep.), collected from fish market, Daliganj. The new species is characterized by shape of body, position of pseudosuckers, hold fast organ and holdfast gland.

15. Metacercaria of Bucephalopsis garuai Verma, 1936 was collected from the gills of Clupisoma garua (Ham.), bought from fish market Chapra (Bihar) is briefly recorded. 173

16. Metacercaria of B. linguiformis Chakrabarti and Baugh, 1974 was collected from liver of Clarias batrachus (Linn.), bought from Kaisarbagh fish market (Lucknow) and is briefly recorded.

17. Metacercaria of Orchipedum Braun, 1901 was collected from the body cavity of Channa punctatus (Bloch) at Sector-3 (Lucknow). The present specimen is the first record of the metacercaria from fish.

18. Metacercariia of Opisthorchis elongatus Agrawal, 1975 was collected from the gills of Esomus danricus (Ham) bought from an aquarium shop (Lucknow) and is briefly recorded.

19. A Plagiorchiid metacercariae was collected from the body cavity of Channa punctatus (Bloch), collected from fish market of Sector-3 (Lucknow) and is briefly recorded.

20. Metacercaria of Ommatobrephus Mehra, 1928 was collected from body cavity of Colisa fasciatus (Bloch. & Schn.), brought to lab from fish market of Sector: 3 (Lucknow) and is redescribed. It is the first record of the metacercaria from India.

21. Metacercaria of Isoparachis hypselobagri (Billet, 1898) Odhner, 1927, was collected from body cavity of Wallago attu (Bloch & Schn.) at Kanpur fish market. It is briefly recorded.

22. Metacercaria of Genarchopsis goppo Ozaki, 1925 was collected from gut of Channa punctatus (Bloch) at Itwara fish market (Bhopal). This appears to be first record of the metacercaria from India.

Part-II

Part-II deals with the description of 16 adult trematodes.

1. Bucephalus bhagalpurensis n. sp. was collected from gut of Mystus tengra (Ham.) at River Saryu Bhagalpur, (Near Belthara, U. P.). The new species is characterized by position of pharynx, short cirrus sac and extension of 174

vitellaria. The position of genital pore further separates the new parasite from other species.

2. Bucephalopsis karvei Bhalerao, 1937 was collected from gut of A. coila (Ham), X. cancila (Ham.) and E. vacha (Ham.), bought from fish market, Kanpur, Banaras and Barabanki, which appear to be new localities for the worm.

3. Bucephalopsis fusiformis Verma, 1936 was collected from gut of Eutropiicthys vacha (Ham) at Madhuban Tal and is briefly recorded.

4. Allocreadium itwarensis n. sp. was collected from gut of Mastacembalus puncalus (Ham.), bought from Itwara fish market, (Bhopal). The new species is characterized by the ratio of suckers, position of receptaculum seminis, extension of vitelline follicles, shape and extension of cirrus sac and position of genital pore.

5. Allocreadium catlai Kakaji, 1969, was collected from body cavity of Colisa fasciatus (Bloch & Schneider) at Kanpur fish market and is briefly recorded.

6. Allocreadium badatalabensis n. sp. was collected from gut of Heteropneustes fossilis (Bloch), caught from Bada Talab (Bhopal). The species is characterized by bipartite vesicula seminalis, position of genital pore, and extension of vitellaria and size of eggs.

7. Eucreadium madhubanensis n. sp. was found in the gut of Eutropiicthys vacha (Ham.), collected from Madhuban fish market, District Mau. The new species is characterized by the ratio of suckers, shape of gonads and extension of vitellaria.

8. Rhynchocreadium dohrighatensis n. sp., was found in the body cavity/gut of Channa marulius (Ham.), caught from river in Dohrighat. The species is characterized by ratio of suckers, ratio of testes and ovary and extension of vitellaria. 175

9. Neopodocotyle dohrighatensis n. sp. was collected from the gut of Channa puctatus (Bloch.), caught from river in Dohrighat. The new species is characterized by ratio of sucker, position of genital pore and size of eggs.

10. Peracreadium bhopalensis n. sp. was found in the gut of Macrognathus aculeatus (Bloch), caught from Bada Tal (Bhopal). The new species is characterized by ratio of suckers, position of genital pore and size of eggs.

11. Genarchopsis goppo Ozaki, 1925, was collected from the gut of Wallago attu (Bl. & Schn.), bought from Itwara fish market (Bhopal) and is briefly recorded.

12. Haplorchoides varanasiensis n. sp. collected from the gut of Eutropicthes vacha (Ham.), bought from a fish market at Varanasi. The new species is characterized by the position of ventrogenital sac, number of spines in ventrogenital sac and extension of vitellaria.

13. Opisthorchis pedicellata Verma, 1927, was collected from gall bladder of Rita rita (Ham.), bought from fish market, Lucknow and is briefly recorded.

14. Oudhia itwaraensis n. sp. was collected from the gut of Heteropneustes fossilis (Bloch), bought from Itwara fish market, (Bhopal). The species is characterized by in ratio of suckers, extention of vitellaria and position of genital pore.

15. Phyllodistomum vachius Dayal, 1949, was found from urinary bladder of Mastacembalus puncalus (Ham.), caught from Bada Talab (Bhopal) and is briefly recorded.

16. Pleuragenoid madhubanensis n. sp. was collected from the gut of Ompok bimaculatus (Bloch), caught from Madhuban Tal. The new species is characterized by ratio of suckers, extension of cirrus and position of gonads and genital pore.