Trichodinid ectoparasites from fresh water fishes of Washim region of Maharashtra, India

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1 Parasitological Communication Biosci. Biotech. Res. Comm. 8(1): (2015) Trichodinid ectoparasites from fresh water fishes of Washim region of Maharashtra, India JR Somatkar* and DS Dabhade Post Graduate and Research Department of Zoology, R. A. Arts, Shri M. K. Commerce and Shri S. R. Rathi Science Mahavidhyalaya, Washim , India ABSTRACT The present study is undertaken to study the morphometrics of ciliated protozoan parasites together with histopathology of freshwater fishes infected with parasitic protozoans. Four ectoparasitic Trichodinids (Ciliophora: Peritrichida) were isolated from freshwater fishes mainly the Indian major carps collected from various water resources in the Washim region of Maharashtra, India. The present research paper emphasizes only the morphometrics of ciliated protozoan parasites found on local fish fauna. The ectoparasitic Trichodinids reported and morphologically described are Tripartiella obtusa, Tripartiella bulbosa, Tripartiella copiosa and Trichodinella epizootica. Taxanomic descriptions of these Trichodinids are based on the structure of the buccal ciliature and on the appearance of the adhesive disc revealed by silver impregnation method (Klein, 1958). During the present study, it was observed that the presence of these ectoparasitic Trichodinid species heavily damage and distruct the gill tissues of fish which in turns results in degeneration of the respiratory surface of fish. KEY WORDS: PARASITES, TRICHODINIDS, TRIPARTIELLA, TRICHODINE-LLA, FRESHWATER FISHES, WASHIM. INTRODUCTION Aquaculture has been an important vector in the introduction, transfer, and spread of some diseases and parasites. Parasite, an important group of pathogen causes infection and diseases of fish both in freshwater and marine environments Klinger and Floyd, 2002, Hemananada (2012). With the increasing interests in aquaculture parasitic infestations are becoming threats for fish health management and aquatic crop production throughout the world. It is therefore an essential area for proper attention to be given by the scientists for sustainable aquaculture production. Ciliated protozoa (Trichodinids) are among the most common external parasites present on the skin and gills of fish (Dickerson and Clark, 1996; Lom, 1995 (Chandra, 2006). The ectoparasitic Trichodinids have been earlier reported by many workers in Indian fishes from various regions. Mukherjee and Haldar (1982) were the first ARTICLE INFORMATION: *Corresponding Author Received 20 th April, 2015 Accepted after revision 28 th June, 2015 BBRC Print ISSN: Online ISSN: NAAS Journal Score : 3.48 A Society of Science and Nature Publication, All rights reserved. Online Contents Available at: http// 35

2 in India, who reported the members of the genus Tripartiella. After that Samal (1987), Sarkar (1988), Saha et al. (1995), Basu and Haldar (1998), Mitra and Haldar (2003), Mohilal and Hemananada (2012) have reported many species of Tripartiella from India. During the present research work, four known species of Trichodinids viz. Tripartiella obtusa, Tripartiella bulbosa, Tripartiella copiosa and Trichodinella epizootica. were reported from freshwater fishes. The Trichodinid parasite species have not been reported earlier from Maharashtra, India and the present research is the first record of the ectoparasitic Trichodinid from Washim in Maharashtra state of India. The present research work has been undertaken to study the parasitic population present on locally available freshwater fishes in order for public understanding and support to make the people aware of the value of fish by improving protective measures by parasitic diseases. MATERIALS AND METHODS The fishes for the present study were collected during January 2009 to July 2013 from the resources such as local fish markets and nearby dams in Washim region of Maharashtra. The live fishes were collected and transported to the laboratory by placing them into well aerated water in plastic bag. Fishes available in the local market were purchased, freezed in ice-box and brought to the laboratory of P. G. and research department of Zoology, R. A. College, Washim for further investigation. In the laboratory, the length and weight of fishes sampled were measured and fishes were subjected to necropsy procedure for parasitological examination. Gill and skin smears were prepared. Air-dried smears were examined under microscope. Slides with trichodinid ciliates were impregnated with Klein s dry sliver impregnation technique (Klien, 1958). Examinations of preparations were made under Olympus phase-contrast microscope at 100x magnification and The photographs of the slide were taken with the camera set on phase contrast microscope and drawing will be made with Camera Lucida. RESULTS AND DISCUSSION The various trichodinid parasites reported during the present study are described as below: 1. Tripartiella obtusa (Ergens and Lom 1970) (Fig. 3-4) Taxonomic summary: Type host: Labeo rohita, Cirrhinus mrigala, Parasite: Type locality: Site of: Holotype: Tripartiella obtusa (Ergens and Lom 1970) Ekburgi dam, Supkhela dam Gills infection In slide no. TR15c/2013, deposited in P.G. and Research department of Zoology, R.A. College, Washim Dist. Washim Maharashtra. Description: 1. Small ciliophoran with a disc shaped body. 2. Concave disc surrounded by a finely striated border membrane. 3. Uniformly dark central area of the adhesive disc. 4. The blade elongated and slanted in a backward direction. 5. The distal margin is rounded with curved tangent point which forms a line with y - 1 axis. 6. The anterior projection resembles a rounded triangle, forms together with a V shaped blade pointed anticlockwise. 7. The part of blade adjoining this projection is stalk like. 8. Posterior projection is ill-developed. 9. Anterior projection fits well into the notch of the preceding denticle and is connected to central part by a well developed, obliquely backward, but straight section. 10. The central part is delicate and bluntly rounded in shape. 11. The ray is stout, straight and short to a lesser extent, directed anteriorly and terminates in a rounded point. 12. The adoral ciliary spiral turns about 190o 200o. Reported by: Ergens and Lom (1970) reported first time from Czechoslovakia Lom and Haldar (1977) from Czechoslovakia. Mohilal and Hemananda (2012) from Manipur. Remarks: (Refer Table 1) Tripartiella obtusa was first reported by Ergens and Lom (1970) from the gills of Gibio gobio in Czechoslevakia. Lom and Haldar (1977) also reported this ciliated parasite from the gill of Gibio gobio in Czechoslovakia. Mohilal and Hemananda (2012) also revealed the presence of the Tripartiella species in freshwater fishes of Manipur. The Tripartiella species reported in the present investigation have identical denticle structure and dimensions with T. obtusa as mentioned by Ergens and Lom (1970), Lom and Haldar (1977) and Mohilal and Hema- 36 TRICHODINID ECTOPARASITES FROM FRESH WATER FISHES OF WASHIM REGION OF MAHARASHTRA, INDIA BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS

PLATE 1: Showing microphotographs of Trichodinid parasites along with drawing. nanda (2012). Hence the present Tripartiella species is identified as Tripartiella obtusa (Ergens and Lom, 1970). 2.

3 PLATE 1: Showing microphotographs of Trichodinid parasites along with drawing. nanda (2012). Hence the present Tripartiella species is identified as Tripartiella obtusa (Ergens and Lom, 1970). 2. Tripartiella copiosa (Lom, 1959) (Fig.1-2) Taxonomic summary: Type host: Labeo rohita, Cirrhinus mrigala, Catla catla Parasite: Tripartiella copiosa (Lom 1959) Type locality: Ekburgi dam, Tornala dam Site of infection: Gill Holotype: In slide no. TR12c/2013, deposited in P.G. and Reserech department of Zoology, R. A. College,Washim Dist. Washim Maharashtra. Description: 1. Small ciliophoran with a disc shaped body. 2. Adhesive disc surrounded by a finely striated border membrane. BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS TRICHODINID ECTOPARASITES FROM FRESH WATER FISHES OF WASHIM REGION OF MAHARASHTRA, INDIA 37

4 Table 1: Morphometric comparison of Tripartiella obtusa obtained in the present study with those of Ergens and Lom (1970), Mitra(2002) and Mohilal and Hemananda (2012). Character T. obtuse T. obtusa T. obtusa T. obtusa Diameter of body Diameter of adhesive disc Dimension of body denticulate ring Width of the border membrane Number of denticles Number of radial pins/ denticle Width of the central part Adoral ciliary spiral Host G. gobio Notopterus notopterus L.rohita L.rohita, Cirrhinus mrigala Site of infection Gills Gills Gills Gills Reference Ergensand Lom(1970) Mitra (2002) Mohilal and Hemananda (2012) Present study 3. The centre of this disc is finely granular and uniformly dark when impregnated with silver nitrate. 4. Denticle small. The blade of denticle is broad, fills half of the space between y axis and obliquely directed backwards. 5. The distal margin in most cases is truncated, adjoins and runs parallel to border membrane. 6. The lateral margins of blade are parallel. The anterior margin is almost parallel to y +1 axis and constricts at the anterior projection. 7. The posterior margin also has the same course and adheres to y - 1 axis throughout its length. 8. The tangent point is flat, forms a line with y-axes. 9. The anterior projection is prominent, but is a delicate spike like extension. 10. The blade is connected to anterior projection by a delicate section. The posterior projection is also prominent and touches the section between blade and anterior projection by its tip. The lateral projection fits well into the well developed notch at the denticle. 11. The section between anterior projection and central part is delicate. 12. The central part is also delicate. 13. The ray is thin, slanted posteriorly terminating in a rounded point. 14. The adoral zone of ciliary spiral takes a turn of about 180o 200o. Reported by Lom(1959) reported for the first time from Czechoslovakia. Lom and Haldar (1977) from Czechoslovakia. Mohilal and Hemananda (2012) from Manipur Remarks (Refer Table 2): Tripartiella copiosa was first reported by Lom, (1959) from the gills of Rhodeus bericeus in Czechoslevakia. Lom and Haldar, (1977) contributed in characterizing T. copiosa as the ciliophoran of the genus Tripartiella. Mohilal and Hemananda, (2012) also revealed the presence of the Tripartiella copiosa in freshwater fishes of Manipur. The Tripartiella species reported in the present investigation have same morphometric features as described by Lom, (1959), Lom and Haldar, (1977) and Mohilal and Hemananda (2012). Hence the present Tripartiella species is identified as T. copiosa Lom, (1959). 3. Tripartiella bulbosa (Davis, 1947; Lom, 1959) (Fig.5-6) Taxonomic summary: Type host: Labeo rohita, Catla catla Parasite: Tripartiella bulbosa (Davis, 1947; Lom, 1959) Type locality: Kajal Amba dam, Ekburgii dam, Tornala dam Site of infection: Gills Holotype: In slide no. TR14c/2013, deposited in P.G. and Research department of Zoology, R.A. College, Washim Dist. Washim Maharashtra. Description 1. Small ciliophoran with a disc shaped body. 38 TRICHODINID ECTOPARASITES FROM FRESH WATER FISHES OF WASHIM REGION OF MAHARASHTRA, INDIA BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS

5 Table 2: Morphometric comparision of Tripartiella copiosa obtained in the present study with those of Lom and Haldar (1977), Mitra (2002) and Mohilal and Hemananda (2012). Character T. copiosa T. copiosa T. copiosa T. copiosa Diameter of body Diameter of adhesive disc Dimension of body denticulate ring Width of the border membrane Number of denticles Number of radial pins/denticle Apr Width of the central part Adoral ciliary spiral Host Rutilus rutilus L.bata C. mrigala L.rohita, Cirrhinus mrigala and C. catla Site of infection Gills Gills Gills Gills Reference Lom and Haldar (1977) Mitra (2002) Mohilal and Hemananda (2012) Present study 2. Adhesive discs dark. 3. Uniquely shaped denticles. 4. The blade is elongated, slanted obliquely backwards with parallel lateral margins, which are constricted at either end, but a sort of constriction is prominent at the mid length of blade. 5. The distal margins of the blades are almost rounded. Tangent point is flat and is situated lower than the distal margin. 6. Anterior projection of the blade is reduced to an insignificant tubercle. 7. Well developed posterior notch is present. 8. The section connecting anterior projection and central part is well formed and curved towards the central area. 9. The central part of the denticle is short, knee like and bent. 10. The ray is short, stumpy and directed towards the centre of adhesive disc. 11. Horse shoe shaped macronucleus observed. 12. The adoral ciliary spiral makes a turn of Reported by: Davis (1947) first described the species from West Virginia, USA. Chen (1956) from China Lom and Haldar (1977) from West Bengal Mukherjee and Haldar (1982) from West Bengal Das and Haldar(1987) from West Bengal Basu and Haldar (1998) from West Bengal Mohilal and Hemananda (2012) from Manipur Remarks (Refer Table 3): Tripartiella bulbosa was first reported by Davis (1947) from the gills of Margariscus margarila in West Virginia, USA. Chen (1956) also reported it from four Chinese herbivorous fishes. Mukherjee and Haldar (1982) reported T. bulbosa from freshwater fishes. Das and Haldar (1987) reported the same from Catla catla and Catla mrigala. Basu and Haldar (1998) reported T. bulbosa from cultured freshwater fishes in West Bengal. Mohilal and Hemananda (2012) also recorded the presence of Tripartiella bulbosa from Indian Major Carps in Manipur. According to Lom and Haldar (1977) T. bulbosa falls into the group of Tripartiella with denticles characterized by having a delicate central part, unconstructed blade at either end with parallel borders joining the centre by a more or less pronounced knee like bend. The parasite obtained in the present investigation have the same identical features mentioned above, Thus, the present parasite is identified as T. bulbosa (Davis 1947; Lom1959). 4. Trichodinella epizootica (Raabe, 1950) (Fig. 7-8) Taxonomic summary: Type host: Labeo rohita, Cirrhinus mrigala, Catla catla Parasite: Trichodinella epizootica Type locality: Ekburgi dam, Supkhela dam Site of infection: Gills Holotype: In slide no. TR11d/2013, deposited in P.G. and Reserech department of Zoology, R.A. College, Washim Dist. Washim Maharashtra. BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS TRICHODINID ECTOPARASITES FROM FRESH WATER FISHES OF WASHIM REGION OF MAHARASHTRA, INDIA 39

6 Table 3: Morphometric comparision of Tripartiella bulbosa obtained in the present study with those of Davis(1947), Das and Haldar (1987) and Mohilal and Hemananda (2012). Character T. bulbosa T. bulbosa T. bulbosa T. bulbosa Diameter of body Diameter of adhesive disc Dimension of body denticulate ring Width of the border membrane Number of denticles Number of radial pins/denticle Width of the central part Adoral ciliary spiral Host M.margarita C. mrigala, C. catla L.rohita, Calta catla and C. mrigala Site of infection Gills Gills Gills Gills Reference Davis(1947) Das and Haldar(1987) Mohilal and Hemananda(2012) L.rohita, C. catla Present study Table 4: Morphometric comparision of Trichodinella epizootica obtained in the present study with those of Lom and Haldar (1977), Mitra and Haldar (2004) and Tao and Zhao (2006). Character T. epizootica T. epizootica T. epizootica T. epizootica Diameter of body Diameter of adhesive disc Dimension of body denticulate ring Width of the border membrane Number of denticles Number of radial pins/denticle May Width of the central part Adoral ciliary spiral Host P. fluviatilis Puntius gelius A. nobilis, H.molitrix L.rohita, Cirrhinus mrigala and C. catla Site of infection Gills Gills Gills Gills Reference Lom and Haldar (1977) Mitra and Haldar (2004) Tao and Zhao (2006) Present study Description: 1. Small ciliated parasite having shape of a vaulted disc. 2. Adhesive disc concave. 3. Narrow border membranes with fine striations. 4. Blade elongated with almost parallel margins and tapering to region of anterior projection. 5. Distal margin closely adjoining border membrane truncated or slightly curved with blunt tangent point which forms small line with y-1- axis, situated lower than distal margin. 6. Anterior margin runs at slight angle to y-axes. 7. Anterior projection slender, but prominent and fits well into notch between central part and blade of preceding denticle, directed obliquely in a distal direction in some specimens. 8. Posterior projection not well developed. 9. Central part rounded and pronounced touches and extends slightly beyond y axes. 10. Macronucleus horseshoe shaped, but micronucleus could not be detected. 11. Adoral ciliary spiral takes a turn of 180. Reported by: Lom and Dykova (1992) from North America. Basson et. al. (1983) from South Africa 40 TRICHODINID ECTOPARASITES FROM FRESH WATER FISHES OF WASHIM REGION OF MAHARASHTRA, INDIA BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS

7 Tao and Zhao (2006) from China Basson (2010) from Australia Remarks (Refer Table 4): T. epizootica was first described by Raabe (1950) from the gills of fishes. Later on Lom and Dykova (1992), Basson et.al,(1983) described the species from Cyprinus carpio from South Africa. Mitra and Haldar (2004) also reported T. epizootica infestation in minor carp, Puntius gelius in India. Tao and Zhao (2006) found T. epizootica on the gills of H. molitrix and Aristichthys nobilis. The morphometric data and shape of denticle of Trichodinid species reported in the present study are consistent with Lom (1961), Mitra and Haldar (2004) and Tao and Zhao (2006), hence it is identified as T. epizootica (Raabe, 1950). CONCLUSION The present study indicates the presence of four species of ectoparasitic Trichodinids on gill surface of freshwater fishes mainly the Indian major carps in greater population which results in debilitary effect on host. So in conclusion controlling measures should be taken to interrupt the steps of parasitic transmission from one host to other. Emphasis should be given to control the parasite with a view to increase the protein production together with the rapid growth of fishes. Therefore extensive study should be carried out on the protozoan parasites otherwise: the pathogenecity caused by them by damaging the tissue and decreasing the nutritional value, will lower the productivity of fish. ACKNOWLEDGEMENTS We are thankful to Principal Dr. M. M. Sancheti, R.A. Arts, Shri M. K. Commerce and Shri S. R. Rathi Science Mahavidyalaya, Washim. (M.S.)., for encouraging research and providing highly equipped laboratory facility. REFERENCES Basson L. (2010). First records of Trichodinid ectoparasites (Ciliophora: Peritrichia) from introduced freshwater fishes in Tasmania, Australia, with comments on pathogenecity. Acta protozool. 49: Basson L., Van As J.G., and Paperna I. (1983). Trichodinid ectoparasites of cichlid and cyprinid fishes in South Africa and Israel. Syst. Parasitol. 5: Basu S. and Haldar D.P. (1998). Comparative study on prevalence of protozoan parasite in pure and hybrid carps. Environ Ecol 16(3): Chandra K.J. (2006). Fish parasitological studies in Bangladesh: a review. J Agric Rural Dev 4(1and2): Chen C.L. (1956). The protozoan parasites from four species of Chinese pond fishes: Ctenopharyngodon idellus, Mylopharyngodon piceus, Aristichthys nobilis and Hypophthalmichthys molitrix II. The protozoan parasites of Mylopharyngodon piceus. Acta Hydrobiol Sin 1: Dickerson H.W. and Clark T.G. (1996). Immune response of fishes to ciliates. Annual Review of Fish Diseases 6: Das M.K. and Haldar D.P. (1987). Urceolariid ciliates of genus Tripartiella invading gills of fresh water cultured carps in India. Arch Protistenk 134: Davis H.S. (1947). Studies of the protozoan parasites of freshwater fishes. U. S. Dep Interior Fish Bull 51:1 29 Golemansky V. and Grupcheva G. (1975): Studies on unicellular parasites of one year old fingerlings of herbivorous fishes in Bulgaria. Acta Zool Bulg 2:3 14 Kashkovsky V.V. (1974): Urceolariids (ciliate, peritricha) from Ural fishes. Parazitologiya. 8: Klien B.M. (1958). The dry silver method and its proper use. J Protozool 5: Klinger R.E. and Francis-Floyd R. (2002). Introduction to Freshwater Fish Parasites, CIR, Institute of Food and Agricultural Sciences, University of Florida: 716. Lom J. and Dykova I. (1992). Protozoan parasites of fishes. Developments in Aquaculture and Fisheries Science, vol. 26: 315. Lom J. (1995). Trichodinidae and other Ciliates (Phylum Ciliophora). Fish Diseases and Disorders: Protozoan and Metazoan Infections. CAB International, Cambridge Lom J. (1959). On the systematic of the genus Trichodinella Sra mek- Husˇek (=Brachyspira Raabe). Acta Parasitol Pol 7: Lom J. and Haldar D.P. (1977). Ciliates of the genera Trichodinella, Tripartiella and Paratrichodina (Peritricha, Mobilina) invading fish gills. Folia Parasitol 24: Mitra A.K. and Haldar D.P. (2003). Record of a trichodinid ciliophoran, Tripatriella copiosa Lom, (Mobilina: Trichodinidae) infesting gills of Mystus vittatus (Bloch) from Canning, South 24 Paraganas. Environ Ecol 21: Mitra A.K. and Haldar D. P. (2004): First record of Trichodinella epizootica (Raabe, 1950) Sramek-Husek, 1953, with description of Trichodina notopteridae sp.n. (Ciliophora: Pertrichida) from freshwater fishes of India. Acta Protozool. 43: Mohilal N. and Hemananda T. (2012). Record of the species of Tripartiella (Lom, 1959) from fishes of Manipur. J Parasit Dis (Jan-June 2012) 36(1): Mukherjee M. and Haldar D.P. (1982). Observations on the urceolariid ciliates of the genera Trichodina and Tripartiella in freshwater teleosts. Arch Protistenkd 126: BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS TRICHODINID ECTOPARASITES FROM FRESH WATER FISHES OF WASHIM REGION OF MAHARASHTRA, INDIA 41

8 Samal K.K. (1987). Observations on the protozoan parasites in the brackish and freshwater edible fishes of Orissa. Ph. D. Thesis, Kalyani University. Saha B.S., Bandhyopadhyay P.K. and Haldar D.P. (1995). Biodiversity of trichodinid ciliates in fresh water fishes of West Bengal. Environ Ecol 13(4): Sarkar S. (1988). Studies in the Urceolariid ciliates (Peritricha,Mobilina) from edible fishes of West Bengal. PhD Thesis, Kalyani University, West Bengal, 250. Tao Y.F. and Zhao Y.J. (2006). Ectoparasitic Trichodinids from some freshwater fishes in the Chongqing area, China, with describtion of a new species of the genus Trichodina Eherenberg, Acta Zootaxonomica Sinica, 31(4): TRICHODINID ECTOPARASITES FROM FRESH WATER FISHES OF WASHIM REGION OF MAHARASHTRA, INDIA BIOSCIENCE BIOTECHNOLOGY RESEARCH COMMUNICATIONS

Protozoologica. Mohamad Ahmad ALI INTRODUCTION. Acta Protozool. (2009) 48:

Protozoologica. Mohamad Ahmad ALI INTRODUCTION. Acta Protozool. (2009) 48: Acta Protozoologica Acta Protozool. (2009) 48: 265 270 Light and Scanning Electron Microscopy of Dipartiella simplex (Raabe, 1959) Stein, 1961 Infesting Common Sole (Solea vulgaris) and Goby Fish (Gobius