TWO NEW SPECIES OF PTYCHOBOTHRIUM LOENNBERG, 1889 (PLATYHELMINTHES: CESTOIDEA) FROM FRESHWATER FISHES OF INDIA

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1 Research Article NBU J. Anim. Sc. 11: (2017) ISSN TWO NEW SPECIES OF PTYCHOBOTHRIUM LOENNBERG, 1889 (PLATYHELMINTHES: CESTOIDEA) FROM FRESHWATER FISHES OF INDIA Suranjana Banerjee¹*, Buddhadeb Manna² and Ashok Kanti Sanyal¹ 1 Zoological Survey of India, M-Block, New Alipore, Kolkata , India 2 Parasitology Research Unit, Department of Zoology, University of Calcutta, India Received: March 27, 2017 Revised & Accepted: November 17, 2017 ABSTRACT The genus Ptychobothrium Loennberg, 1889 contains 10 valid species. In this paper, two new species, Ptychobothrium leiodoni sp.n. and Ptychobothrium raiamusi sp.n. are described from the freshwater fishes, Leiodon cutcutia Hamilton, 1822 and Raiamus bola Hamilton, 1822 belonging to West Bengal and Systomus sarana Hamilton, 1822 and Raiamus bola Hamilton, 1822 belonging to Bihar, respectively. The hosts were purchased from a local fish market. Ptychobothrium leiodoni sp.n. is characterized by a small, inverted heart-shaped scolex, measuring mm in length and mm in breadth; testes in number and, round in shape and a transversely elongated ovary while Ptychobothrium raiamusi sp. n. is characterized by body measuring mm in length and mm in breadth; small, inverted heart-shaped scolex measuring mm in length and mm in breadth, testes numbering and a transversely elongated ovary that differentiates it from the rest of the described species in the genus. Keywords: Ptychobothrium leiodoni sp.n., Ptychobothrium raiamusi sp.n., Leiodon cutcutia, Systomus sarana, Raiamus bola, West Bengal, Bihar, India. INTRODUCTION The genus Ptychobothrium was created by Loennberg in 1889 under the family Ptychobothriidae Luhe, 1902 and it was selected as the type genus for that family by Loennberg in The genus Ptychobothrium Loennberg, 1889 is presently placed under the family Bothriocephalidae Blanchard, Khalil et al. (1994) synonymized the family Ptychobothriidae Luhe, 1902 with the family Bothriocephalidae Blanchard, The type species of Ptychobothrium is Ptychobothrium belones Dujardin, Ptychobothrium belones was recorded by Dujardin (1845) as Bothriocephalus belones from the marine fish, Belones acus at Cette, France. Loennberg (1889) made Dujardin s species the genotype of the new genus Ptychobothrium. Linton (1891) reported it as Dibothrium restiforme from Tylosurus carribaeus, Woods Hole. Linton (1934) described *Author for correspondence: Dr. S. Banerjee, E. Mail: serenebanerjee@gmail.com small immature worms from Exocoetus volitans as Bothriocephalus sp. and its detailed description was published by Janicki (1928). It was also found by Ariola (1896) in Belone vulgaris. Yamaguti (1934) redescribed it from Tylosurus shismtorhynchus in Japanese waters. Shuler (1938) recorded it from Strongylura notata and Tylosurus raphidoma at Dry Tortugas, Florida. In India, six species of Ptychobothrium have been described from freshwater fish hosts and only one species, Ptychobothrium cypseluri was recorded by Rao (1959) from a marine teleost Cypselurus poecilopterus, off the Waltair coast in Andhra Pradesh from the Bay of Bengal. In course of a study tour to collect cestode parasites from fish hosts from West Bengal and Bihar, three cestode parasites belonging to the genus Ptychobothrium Loennberg, 1889 were recovered from freshwater fishes Leiodon cutcutia Hamilton, 35

2 Suranjana Banerjee, Buddhadeb Manna and Ashok Kanti Sanyal 1822, Systomus sarana Hamilton, 1822, Raiamus bola Hamilton, The genus Ptychobothrium Loennberg, 1889 has been reported only from fish hosts in India. The proposed new cestode Ptychobothrium leiodoni sp.n. from the freshwater fishes Leiodon cutcutia and Raiamus bola has been added in the present work from West Bengal. Ptychobothrium raiamusi sp.n. collected from the two fish hosts Systomus sarana and Raiamus bola is described and proposed to be new to science from Bihar. This is also the first report of the genus Ptychobothrium Loennberg, 1889 from both the states of West Bengal and Bihar, India. MATERIALS AND METHODS One cestode parasite of the proposed new species Ptychobothrium leiodoni sp.n. was recovered from the intestine of a freshwater fish Leiodon cutcutia from Bongaon, North 24- Parganas, West Bengal and another single cestode parasite was collected from the intestine of Raiamus bola from Mynaguri, West Bengal in the month of May, On close examination, both the collected cestode parasites revealed the same morphological characters though they belonged to two different hosts. One cestode parasite of the proposed new species Ptychobothrium raiamusi sp.n. was collected from the intestine of Raiamus bola and another single cestode parasite having the same morphological characters was collected from the intestine of Systomus sarana from Bodh Gaya, Bihar in the month of July, The parasites were fixed, stained, observed and identified following standard techniques. The parasite specimens were fixed and preserved in 70% alcohol. They were then stained in alcoholic borax carmine and mounted on slides in Canada balsam. The specimens were studied under microscope (Magnus MLX-Trinoculor from Olympus, Japan) and camera lucida drawings and photomicrographs were made as per need to describe the species. All the measurements given in the text are in millimetres unless otherwise stated. RESULTS 1. Ptychobothrium leiodoni sp.n. (Fig.1 and 2): Diagnosis: The body is divided into many immature, mature and gravid proglottids with complete and incomplete segmentations. The immature proglottids measure in length and in breadth. The mature proglottids measure in length and in breadth. However, the gravid proglottids were not present. The parasite measures in length and in breadth. The scolex is well-developed, inverted heart-shaped with a rounded anterior end. Each lateral surface of the scolex has a furrow continuous with that of the proglottid and is not folded longitudinally but has smooth margins. The scolex measures in length and in breadth. The two bothria are dorsal and ventral in position, well-developed and present a more or less H-shaped appearance. Each bothridium measures in length and in width. Neck is short and measures 0.11 in length and 0.2 in breadth. The testes are rounded, measuring to 0.01 in length and 0.01 in breadth, in number, lie in the dorsal medulla in two lateral groups and are densely arranged. In mature proglottids, the testes are preovarian, postovarian and are also present on the lateral side of the ovary. The cirrus sac is small, elongated and is transversely placed, slightly to the left side of the median line of mature and immature segments. The cirrus sac measures in length and in breadth. It opens through the genital pore in the middle of the segment. The genital pores are irregularly alternate. The cirrus sac opens through the common genital pore into the genital atrium in front of the ovary. The ovary is transversely elongated, curved anteriorly and present in the posterior half of the segment. In immature proglottids, the ovary measures in length and in breadth. In mature proglottids, the ovary measures in length and in breadth. The oviduct opens into the seminal receptacle. The vagina extends upto the genital pore 36

3 Two new species of Ptychobothrium from freshwater fish Fig. 1 Camera lucida drawings of Ptychobothrium leiodoni sp.n. a Scolex b Immature proglottids c Mature proglottids d Gravid proglottids Fig. 2 Photomicrographs of Phychobothrium leiodoni sp.n. a Scolex b Immature proglottids c Mature proglottids d Gravid proglottids 37

4 Suranjana Banerjee, Buddhadeb Manna and Ashok Kanti Sanyal and is a curved tube. It opens into the genital atrium through the common cirro-vaginal pore which measures 0.04 in length and 0.03 in breadth. The uterus is sac-like, occupying the entire medullary region of the proglottid. It is filled with oval, non operculated eggs. The uterine duct coils forward and is roughly S-shaped in appearance. It opens ventrally through a large uterine pore towards the anterior margin of the segment, above the common genital pore in the median line. The uterine pore measures 0.05 in length and 0.02 in breadth. The vitelline follicles are oval in shape and intermingle with the testes in two lateral fields. The eggs are numerous, oval, thin-shelled and non-operculated, measuring 0.01 in length and in breadth. The longitudinal osmoregulatory canals are two in number. Type Host: Leiodon cutcutia Type Locality: Bongaon, North 24 Parganas, Mynaguri, West Bengal, India. Location: Intestine Dates: and No. of specimens: Two, mounted on two slides. Holotype: One specimen in one slide under the Zoological Survey of India Accession number W9923/1 Paratype: One specimen in one slide under the Zoological Survey of India Accession number W9924/1(Host: Raiamus bola). Deposition: The type specimens having the accession numbers W9923/1 and W9924/1 have been deposited in the Platyhelminthes Section, Zoological Survey of India, New Alipore, Kolkata. Etymology: Considering all these differences in characteristics from all the valid species of the genus, the present species is considered new to science and the authors suggest its name as Ptychobothrium leiodoni sp. n. named after the host Leiodon cutcutia from where the parasite had been recovered. 2. Ptychobothrium raiamusi sp.n. (Figs. 3 and 4): Diagnosis: The body is divided into many immature, mature and gravid proglottids, with incomplete segmentations. Secondary segmentation is present. Mature proglottids are broader than long but gravid proglottids are longer than broad. The immature proglottids measure in length and in breadth. The mature proglottids measure in length and in breadth. The parasite measures in length and in breadth. The scolex is well-developed, inverted heart-shaped with rounded anterior end. Each lateral surface of the scolex has a furrow continuous with that of the proglottid and is not folded longitudinally but has smooth margins. The scolex is remarkably small measuring in length and in breadth. The two bothria are dorsal and ventral in position, well-developed and present a more or less H-shaped appearance. Each bothridium measures 0.21 in length and in breadth. Neck is absent. The testes are rounded, in number, measure in length and 0.01 in breadth and lie densely arranged in the dorsal medulla in two lateral groups. In mature and gravid proglottids the testes are preovarian, postovarian and also present in the lateral sides of the ovary. The cirrus sac is small, elongated and is transversely placed, slightly to the left or right side of the median line of mature and immature proglottids. The cirrus sac measures in length and in breadth. It opens through the genital pore in the middle of the proglottid. The genital pores are irregularly alternate. The cirrus sac opens into a common genital atrium in front of the ovary. The ovary is transversely elongated, curved anteriorly and present in the posterior half of the proglottid. In immature proglottids, the ovary measures in length and in breadth. In mature proglottids, the ovary measures in length and in breadth. The oviduct opens into the seminal receptacle. The vagina arises from the 38

5 Two new species of Ptychobothrium from freshwater fish Fig. 3 Camera lucida drawings of Ptychobothrium raiamusi sp.n. a Scolex b Mature proglottids c Gravid proglottids Fig. 4 Photomicrographs of Ptychobothrium raiamusi sp.n. a Scolex b Mature proglottids c Gravid proglottids 39

6 Suranjana Banerjee, Buddhadeb Manna and Ashok Kanti Sanyal ovary and extends up to the genital pore and is a curved tube. It opens into the genital atrium through the common cirro-vaginal pore which measures 0.05 in length and 0.03 in breadth. The uterus is sac-like occupying the entire medullary region of the proglottid. It later fills the entire proglottid with oval non-operculated eggs. The uterine duct coils forward and is roughly S-shaped in appearance. It opens ventrally through the uterine pore towards the anterior margin of the segment, above the common genital pore in the median line. The uterine pore measures 0.08 in length and 0.09 in breadth. The vitelline follicles are oval and intermingle with the testes in two lateral fields. The eggs are numerous, oval, thin-shelled, non-operculated, measuring in length and 0.01 in breadth. Type Host: Raiamus bola Type Locality: Bodh Gaya, Bihar, India Location: Intestine Date: No. of specimens: Two in two slides Holotype: One in one slide under the Zoological Survey of India Accession number W9925/1 (Host: Raiamus bola) Paratype: One in one slide under the Zoological Survey of India Accession number W9926/1 (Host: Systomus sarana) Deposition:The type specimens having the accession numbers W9925/1 and W9926/1 have been deposited in the Platyhelminthes Section, Zoological Survey of India, New Alipore, Kolkata. Etymology: Considering all these differences in characteristics from all the valid species of the genus, the present species is considered new to science and the authors suggest its name as Ptychobothrium raiamusi sp.n. named after its host Raiamus bola from which the parasite had been recovered. DISCUSSION Currently, the following valid species (Schmidt, 1986) collected from fish hosts are present under the genus Ptychobothrium Loennberg, They are listed as follows: 1. Ptychobothrium belones (Dujardin, 1845) Loennberg, 1889 is the type species of this genus, which was collected from Belone sacus in France. 2. Ptychobothrium cypseluri Rao, 1959 from Cypselurus poecilopterus, off the Waltair coast in Andhra Pradesh from Indian waters. 3. Ptychobothrium khami Shinde and Deshmukh, 1975 from Nemacheilus botia, Chela phulo and Chela clupeoides from Aurangabad in Maharashtra, India. 4. Ptychobothrium phuloi Shinde and Deshmukh,1975 from Nemacheilus botia, Chela phulo and Chela clupeoides from Aurangabad in Maharashtra, India. 5. Ptychobothrium chelai Shinde and Deshmukh, 1976 from Nemacheilus botia, Chela phulo and Chela clupeoides from Aurangabad in Maharashtra, India. 6. Ptychobothrium ratnagiriensis Deshmukh and Shinde, 1976 from a marine fish Exocoetus bahiensis from Ratnagiri, Maharashtra. 7. Ptychobothrium clupeoidesii Chincholikar et al. (1976) from Chela clupeoides collected from Aurangabad, Maharashtra, India. 8. Ptychobothrium nayarensis Malhotra, 1983 from Barilius bola and Schizothorax richardsonii, from river East and West Nayar in Pauri Garhwal District of Uttar Pradesh, India. 9. P. rojanapaibuli Wongsawad, 1998 collected from Mystacoleucus marginatus at Maesa Stream, Chiang Mai, Thailand. 10. P. mystacoleucysi Wongsawad, 1998 collected 40

7 Two new species of Ptychobothrium from freshwater fish from Mystacoleucus marginatus at Maesa Stream, Chiang Mai, Thailand. The proposed new species Ptychobothrium leiodoni sp.n. measures in length and in breadth; scolex small, inverted heartshaped, long and broad; neck short; testes round, in number arranged in two lateral groups in the lateral medulla; cirrus sac small, obliquely placed on the left or right side of the median line, measures in length and in breadth; ovary transversely elongated; uterine sac round; uterine pore located on the midline measures 0.05 in length and 0.02 in breadth; genital pores located in the middle of the proglottid, posterior to the uterine pore. The observed species comes closer to P. mystacoleucysi Wongsawad, 1998 in having similar shape of scolex and number of testes. The observed species differs from P. belones (Dujardin, 1845) Lonnberg, 1889 in which the scolex is spatulate and elongated, more than 3 in length; testes in number and ovary is small and compact; from P. cypseluri Rao, 1959 in which the scolex is arrowhead-shaped, in length; testes in number, ovary triangular and lobulated; uterine sac V-shaped; from P. khami Shinde and Deshmukh, 1975 in which the scolex is 0.91 in length and 1.15 in breadth; testes in number; ovary is cylindrical with small sized acini; cirrus sac measures 0.09 to 0.13 in length and in breadth; from P. phuloi Shinde and Deshmukh,1975 in which the scolex is in length and 1.33 in breadth; testes in number; ovary cylindrical with broad and elongated acini; from P. chelai Shinde and Deshmukh,1976 in which the scolex has a length of 1.57 and breadth of 1.69; testes are in number; ovary is cylindrical with broad acini; from P. ratnagiriensis Shinde and Deshmukh,1976 in which the scolex is spatulate, 3.13 in length and 2.78 in breadth; testes in number; ovary is compact and lobulated; from P. clupeoidesii Chincholikar et al in which the scolex has a length of 1.06 and breadth of 1.27; testes in number; ovary is U-shaped; from P. nayarensis Malhotra,1983 in which the scolex is inverted heartshaped with an apical disc, scolex 1.22 in length and in breadth; testes in number; cirrus sac measures in length and in breadth; ovary V-shaped; from P. mystacoleucysi Wongsawad, 1998 in which the scolex is triangular; testes in number, preovarian; cirrus sac is placed in the centre; vagina is a long tube; ovary bilobed with acini; uterus is reticulate filled with many eggs; uterine pore is rounded placed near to the anterior margin of the proglottid, vitellaria granular; from P. rojanapaibuli Wongsawad, 1998 in which the scolex is heart-shaped; bothridia compressed; immature segments are square-shaped; testes in number, big and round in shape, arranged in two fields, post ovarian; cirrus sac located near the anterior margin of the proglottid, cirrus is curved; uterine pore does not touch the anterior margin of the proglottid; ovary U-shaped; uterus reticulate and vitellaria arranged in 3-4 strip and from another new species described in the present work P. raiamusi sp.n. in which body measures in length and in breadth; scolex small in size, inverted heart-shaped, in length and in breadth; neck absent; testes arranged in two lateral bands, in number; cirrus sac small, transversely placed on the median line, in length and in breadth. The proposed new species Ptychobothrium raiamusi sp.n. measures in length and in breadth; scolex small in size, inverted heartshaped, in length and in breadth; neck absent; testes arranged in two lateral bands, in number; cirrus sac small, transversely placed on the median line, in length and in breadth; ovary transversely elongated; genital pores median; genital atrium present; seminal receptacle present; uterine sac round in shape; vitellaria in two lateral fields. 41

8 Suranjana Banerjee, Buddhadeb Manna and Ashok Kanti Sanyal The observed species comes closer to P. clupeoidesii Chincholikar et al., 1976 in shape of the scolex, absence of neck, number of testes and position of the uterine pore. The observed species differs from P. belones (Dujardin, 1845) Loennberg, 1889 in which the scolex is spatulate and elongated, more than 3 in length; testes in number and ovaryis small and compact; from P. cypseluri Rao, 1959 in which the scolex is arrow-headed, in length; testes in number, ovary triangular and lobulated; uterine sac V-shaped; from P. khami Shinde and Deshmukh,1975 in which the scolex is 0.91 in length and 1.15 in breadth; neck present; immature proglottids measure 0.66 in length and 0.96 in breadth; mature proglottids measure in length and 1.32 in breadth; gravid proglottids measure 2.33 in length and 1.57 in breadth; cirrus sac measures in length and in breadth; testes in number; ovary is cylindrical with small sized acini; from P.phuloi Shinde and Deshmukh, 1975 in which the scolex is in length and 1.33 in breadth; neck present; testes in number; ovary cylindrical with broad and elongated acini; mature proglottids measure in length and in breadth; from P. chelai Shinde and Deshmukh,1976 in which the scolex has a length of 1.57 and breadth of 1.69; testes in number; ovary cylindrical with broad acini; mature proglottids measure 2.68 in length and 2.7 in breadth; from P. ratnagiriensis Shinde and Deshmukh, 1976 in which the scolex is spatulate, 3.13 in length and 2.78 in breadth; neck present; testes in number; ovary compact and lobulated; mature proglottids measure 0.86 in length and 1.73 in breadth; from P. clupeoidesii Chincholikar et al., 1976 in which the scolex has a length of 1.06 and breadth of 1.27; testes in number; ovary U-shaped; mature proglottids are 1.06 long and 1.89 wide; from P. nayarensis Malhotra,1983 in which the scolex is inverted, heart-shaped with an apical disc, scolex 1.22 in length and in breadth; testes in number; cirrus sac measures in length and in breadth; ovary V-shaped; mature segments measure in length and in breadth; from P. mystacoleucysi Wongsawad, 1998 in which the scolex is triangular ; testes in number, preovarian; cirrus sac placed in the centre; vagina is a long tube; ovary bilobed with acini; uterus reticulate and filled with many eggs; uterine pore rounded placed near to the anterior margin of the proglottid; vitellaria granular; from P. rojanapaibuli Wongsawad, 1998 in which the scolex is heart-shaped; bothridia is compressed; immature proglottids are square-shaped; testes in number, big and round in shape, arranged in two fields, post ovarian; cirrus sac located near the anterior margin of the proglottid; cirrus is curved; ovary U-shaped; uterine pore does not touch the anterior margin of the proglottid; uterus reticulate and the granular vitellaria is arranged in 3-4 strips and from another new species described in the present work P. leiodoni sp.n. in which body measures in length and in breadth; scolex small, inverted heart-shaped, long and broad; short neck present; testes rounded, in number; cirrus sac small, measures in length and in breadth. Key to the species of the genus Ptychobothrium Loennberg, Scolex more than 3 in length Scolex less than 3 in length Testes in number... P. belones (Dujardin, 1845) Loennberg, Scolex 3.13 in length, testes in number... P. ratnagiriensis Deshmukh and Shinde, Scolex arrow-shaped... P. cypseluri Rao, Scolex less than 3 in length, inverted heartshaped Scolex less than 3 in length, inverted heartshaped, testes more than

9 Two new species of Ptychobothrium from freshwater fish -Scolex less than 3 in length, inverted heartshaped, testes less than Scolex 1.03 in length, inverted heart-shaped, testes in number, short neck present... P. phuloi Shinde and Deshmukh, Neck absent Scolex 1.22 in length, inverted heart-shaped; testes in number; apical disc present in scolex... P. nayarensis Malhotra, 1983 Apical disc absent in scolex Scolex in length, inverted heart-shaped; apical disc absent; testes in number; ovary transversely elongated... P. leiodoni sp.n. Ovary otherwise Scolex 1.06 in length, inverted heart-shaped; testes in number; Ovary U-shaped, neck absent... P. clupeoidesii Chincholikar et al., Scolex in length, inverted heartshaped; testes in number; ovary transversely elongated, neck absent... P. raiamusi sp.n. 9. Ovary cylindrical ovary not cylindrical Scolex 0.91 in length, inverted heart-shaped; testes in number; ovary cylindrical with small acini... P. khami Shinde and Deshmukh, Scolex 1.57 in length, inverted heart-shaped; testes in number, ovary cylindrical with broad and elongated acini... P. chelai Shinde and Deshmukh, Scolex less than 3 in length, inverted heart shaped; testes in number Scolex less than 3 in length, triangular; testes in number, Ovary U shaped... P. rojanapaibuli Wongsawad, Bilobed ovary with acini... P. mystacoleucysi Wongsawad, 1998 ACKNOWLEDGEMENTS The authors are thankful to the Director, Zoological Survey of India for providing necessary facilities during the course of the work. The authors express their deep thanks to Library Division of Zoological Survey of India, Kolkata HQ for providing necessary facilities. CONFLICT OF INTEREST Declared none by the authors. REFERENCES Ariola, V Sulla Bothriotaenia Plicata (Rud.) e sulsuosviluppo. Atti. Soc. Ligust. Sc. Nat. Geogr. Genova 7: Chincholikar, L. N A new cest ode Ptyc hobothrium cl upeoide sii (Cestoda: Pseudophyllidea) from a freshwater fish from Aurangabad, India. Nat. Sci. J. Marath. Univ. Aurangabad 15(8): Deshmukh, R.A. & Shinde, G.B On Ptychobothrium ratnagiriensis n. sp. from flying fish Exocoetus bahiensis caught off Ratnagiri (West Coast of India). J. Ind. Bios. Assoc. 1: Dujardin, F Histoire Naturelle des Helminthes ou Vers Intestinaux, Paris: xvi pp. Hamilton, F An account of the fishes found in the river Ganges and its branches. Edinburgh & London: i-vii , Pls Janicki, C Cestodes s. str. Aus Fischen und Amphibien. Results of the Swedish zoological expedition to Egypt and the White Nile 1901 under the direction of L.A. Jagerskiold. The Library of the Royal University of Uppsala, Uppsala, Sweden: Loennberg, E Bidrag till kannedomenom i Sverigeforekommande Cestoder (in Danish). Bihang. K. Sven. Vet. Akad. Handl.14:

10 Suranjana Banerjee, Buddhadeb Manna and Ashok Kanti Sanyal Luhe, M Ueber die Fixierung der Helminthenan der Darmwandun gihrer Wirt e und die dadurchverursachtenpathologisch-anatomischen Veranderungen des Wirtsdarmes. Verhandl. Int. Zool. Congr. Berlin: Linton, E A pseudophyllidean cestode from a flying fish. Trans. Am. Microsc. Soc. 53: Linton, E Notes on Entozoa of marine fishes with descriptions of new species. III, Ann. Rep. U.S. Comm. Fish Fisheries, Washington D.C. 16: Malhotra, S. K Cestode Fauna of Hill - Stream Fishes in Garhwal Himalayas, India. VI. Ptychobothrium nayarensis n.sp. from Barilius bola (Ham.) and Schizothorax richardsonii (Gray). Korean J. Parasitol. 21(2): Rao K.H On Ptychobothrium cypseluri n.sp. (Cestoda: Pseudophyllidea) from the flying fish, Cypselurus poecilopterus caught off Waltair. J. Helminthol. 33: Schmidt, G. D CRC Handbook of Tapeworm Identification, CRC Press Inc. Boca Raton Florida: Shinde, G.B. & Deshmukh, R.A On a new cestode Ptychobothrium chelai (Cestoda: Pseudophyllidea) from freshwater fishes from Maharashtra, India. J. Ind. Bios. Asso. 1 (7-9): Shinde, G.B. and Deshmukh, R.A On two new cestodes Ptychobothrium khami and Ptychobothrium phuloi (Cestoda: Pseudophyllidea) from freshwater fishes from Maharashtra, India. J. Ind. Bios. Asso. 1 (7-9): Shuler, R. H Some cestodes of fish from Tortugas, Florida. J. Parasitol. 24: Wongsawad, C A review of th e genus Ptychobothrium Loennberg, 1889 with two new speices. Rivista di Parassitol. 15 (L IX). 3: