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Historical review of piscine trypanosomiasis105Pleomorphism is expressed in the following features: Pleomorphism was observed in T. singhii Gupta and Size changesJairajpuri, 1981b; T. attii Gupta and Jairajpuri, 1982b(small, intermediate and large forms); T. colisi Gupta, Changes in length / width ratio1986 (stumpy, intermediate and slender forms), T.barielliana Gupta , Gupta and Yadav 1987 (slender, The number, width and depth of the waves of the intermediate and large forms) and T. ticti (small,undulating membraneintermediate and large forms) from Puntius ticto The presence or absence of a distinct karyosomein the nucleus(unpublished data of author; Fig. 2)EPresence and number of stainable cytoplasmicgranulesLength of the free end of the flagellumDistance of the kinetoplast from the posteriorendShifts in the position of the nucleus in the body Presence of pellicular striation (“ myonemes ”)in stained preparations.ADPleomorphism in Trypanosoma is well marked inhaving a markedly variegated shape and six differentstages in varying combinations at various periods ofthe digenetic life cycle (vertebrate and invertebratehosts) may occur (trypomastigote, amastigote,promastigote, sphaeromastigote and metacyclicstages). However, trypomastigotes are thepredominant stages reported from fish blood, but inthe vertebrate blood too, pleomorphism may bevisible. Pleomorphism has been reported by someauthors without assigning any names (Becker andOverstreet, 1979; Joshi, 1979) whereas others havegiven specific names to the various forms.Laveran and Mesnil (1907) distinguished the 'large'and 'small' forms of T. remarki; Tanabe (1925) Type I,II and III; Dutton et al. (1907) small, medium andlarge; Qadri (1955) small and large forms of T. striati;Qadri (1962a) large and stumpy forms of T. batrachi;Becker (1967) slender and broad forms of T.occidentalis; Joshi (1982) short, elongated andstumpy forms of T. aori.Pleomorphism in trypanosomes has also beenrecorded from the fishes of Rohilkhand region of UttarPradesh. Dimorphism with two distinguishable formswas recorded in T. aligaricus; Gupta and Jairajpuri,1982a (small and large forms); T. artii Gupta et al.,2002 (small and large forms); T. saulii Gupta et al.,2006 (minuta and magna forms) and T. heteropneustiiGupta et al., 2006 (small and large forms).B10 µmFig. 2. A E. Diagrams showing pleomorphism in Trypanosomaticti from the blood of Puntius ticto. A and B, small forms C andD, intermediate forms; E, large form.BLOOD COLLECTION AND STUDY OF LIVEPARASITESTrypanosomes can be visualized alive in fresh blood.Blood can be collected in several ways but a few dropscan easily be obtained by incising two or three rays atthe base of the caudal fin. The sample of blood can betaken from the heart using a thin Pasteur pipetteintroduced through the ventral body wall or below thepectoral fins (Lom and Dykova, 1992). The clottingtime for fish blood is much shorter than formammalian blood and is to be thus handledaccordingly. A heparinized pipette or a pipettecontaining a droplet of a heparin-saline solution isgenerally recommended. For large fish, blood can betaken directly from the heart. The fresh preparation(ordinary or hanging drop) is immediately examinedfor the presence of parasites which can be seenC
106Guptaactively wriggling about, pushing the blood cells entry to their vector, haematophagous leaches. Afterrandomly during their course of movement. The re-entry into the fish, they may either directly invadeaddition of citrated salt solution prevents blood (or be injected by the vector into) the blood or theycoagulation and leaves the motility of the parasites may enter it after an initial period of multiplication inunimpaired. tissue fluid local to the point of entry (Baker, 1976).During live condition, the parasite moves veryquickly, actively displacing the adjacent red bloodcorpuscles; wriggling movements and twisting of thebody into knots has also been observed occasionally(Qadri, 1962b). Occasionally, the parasites maydisplace themselves by a directional motion.Low intensity infection detectionLom and Dykova (1992) divided the development ofthe parasite in the fish host into four different phases:a) Phase I- Prepatent period (2-9 days)characterized by the absence of flagellates in theperipheral blood. It is not exactly known wherethe organisms are located during this phase andno tissue stages have been reported.Often trypanosomes are present in low intensities and b) Phase II- Appearance of slender forms of thetheir detection in routine blood smears is time flagellates in the peripheral blood- patent andconsuming and the parasites maybe overlooked. Thus increasing parasitaemia due to the division of thelow infection maybe detected in the following trypanosomes.manners:The division of the parasite in fish blood has beenA. Haematocrit centrifuge method (Woo, 1969) reported rarely, it was presumed by some authors thatthe parasites did not multiply in fish blood (Letch,A heparinized capillary tube is filled with about 0.061977). Occasional reports of divisional stages sprungml blood and one end sealed. The tube is centrifugedup without providing clear evidence of the sequentialfor 4 min. at 12,000 rpm. This tube is then placedpattern of division. Laveran and Mesnil (1907)horizontally in a drop of immersion oil on areported that fish trypanosomes multiply bymicroscope slide and examined at 100x magnificationlongitudinal binary fission and protoplasmic massesin a compound microscope. Trypanosomes, if presentundergoing division and young trypanosomes wereare found writhing at the junction of the buffy layerobserved in the blood of Cyprinus carpio and Esox(layer of packed white blood cells) and the clearlucius. Tanabe(1925) observed the dividing formsplasma. Alternatively, the tube is cut immediatelyonly once in the blood of leeches but these forms wereabove this layer, material transferred to a slide, smearnot described. Dividing nuclei in T. maguri (Tandonmade and stained in Giemsa's stain. Concentratedand Joshi, 1973) and T. mukasai (Sinha, 1986) andparasites may be visible under the microscope afterdividing kinetoplast in T. colisi (Gupta, 1986) havecentrifugation (Fig. 4c).been reported. Gupta and Saraswat (1991) observedB. Clot method (Lom and Dykova, 1992)dividing forms in T. rohilkhandae and binucleatedstages were the most common, flagellar division wasTrypanosome infection of extremely low intensity can rare.be detected if blood is allowed to clot in a centrifugetube placed overnight in a refrigerator. The next day, However, the complete sequential pattern of divisionthe flagellates can be found wriggling in the serum of T. danilewskyi in Carassius auratus was explainedoutside the blood clot and may be concentrated by by Woo (1981a). The author divided the stages intocentrifugation. If large amounts of blood are available, four phases arbitrarily : Stage I (production of a newthe trypanosomes may be separated in a DEAE anteriorly directed flagellum and kinetoplastcellulose column (Lanham and Godfrey, 1970; division), Stage II (enlargement and rounding ofLumsden et al., 1973), which is a more complicated posterior end and flagellar shift to posterior end ),method.Stage III (weakly formed undulating membrane andinitiation of nuclear division ) and Stage IV (two fullyMORPHOGENESIS IN THE FISH HOSTformed trypanosomes joined at their posterior end,transverse constriction between the two kinetoplasts,Trypanosoma are truly haematozoic extracellularformation of young parasites with weakly developedparasites spending substantial part of their life cycle inundulating membrane and the nucleus situated closethe plasma. Fish trypanosomes use blood as a route of
Page 1 and 2: TISSN: 0971-7196Journal ofVolume 30
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Page 9 and 10: 100Tandonthe Lifetime achievement a
Page 11 and 12: 102Guptablood of a large variety of
Page 13: 104Guptaused without a proper under
Page 17 and 18: 108Guptaindicate that the pattern o
Page 19 and 20: 110Guptaas several months after fee
Page 21 and 22: 112GuptaBrumpt E. 1906. Sur quelque
Page 23 and 24: 114GuptaLom J, Dykova I and Machako
Page 27 and 28: 118Singhfalciparum-infected human e
Page 29 and 30: 120 Singhmalarial antigens to T-lym
Page 31 and 32: 122 Singhmacrophage tumoricidal act
Page 33 and 34: 124 SinghTaliaferro WH and Connam P
Page 35 and 36: 126Chakrabarti and Mannathe present
Page 37 and 38: 128Chakrabarti and MannaFig. 2a. N.
Page 39 and 40: 130Chakrabarti and MannaAccession /
Page 41 and 42: 132Chakrabarti and Mannamosquito (D
Page 45 and 46: 136Vijayalakshmi and Ramalingam(a)(
Page 49 and 50: 140Raut et al.Table II. Ectoparasit
Page 53 and 54: 144Ravindran et al.RESULTSThe speci
Page 57 and 58: 148WakidKinyoun's modified techniqu
Page 59 and 60: 150 WakidTable I: Food handlers' na
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Page 63 and 64: 154Radha et al.observations have re
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156Radha et al.electron-lucid core
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158Radha et al.(a)(b)(c)(d)(e)Fig.
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160Radha et al.(a)(b)(c)(d)Fig. 4.
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162Radha et al.Morseth DH. 1966. Th
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164Choubisa and ChoubisaMATERIALS A
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166Choubisa and Choubisahelminthic
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Journal of Parasitic Diseases: Dece
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170Pramanik and Manna(a)(b)Fig. 2.
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174Das et al.Valsala KV and Bhaskar
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176Hirani et al.Table I. Incidence
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180Ravindran et al.parasitaemia. Ne
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182Aggarwal et al.positivity indica
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186Bapna et al.Fig. 3: Posterior en
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188 ObituaryHe published more than
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J P DAUTHOR INDEX (2006)Agrawal, MC
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J P DLIST OF REFEREES (2006)Dr. S.
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Ultrastructure, differential densit
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