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P a R T . III. STUDIES on Hepatozoon in SOME SNAICES

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P a R T . III. STUDIES on Hepatozoon in SOME SNAICES PART. III. STUDIES ON HepatOZOOn IN SOME SNAICES 140 INTRODUCTION Hepatozoon is an intra-corpuscular non-pigmented parasite of vertebrates. Levine,et al»,(1980) placed Haemogregarina Danilewsky, 1885, Hfepatozoon Miller, 1908 and Karyolysus Labb^,1894 in three distinct: families viz,, Haemogregarinidae Leger,1911,„ Hepatozoidae WenYon,1926 and Karyolysidae Wenyon,1926 of the suborder Adeleina under the Class Sporozoea, The characteristic features of the genus Haemogregarina Danilewsky,1885 has • been given in Part II, page nimber. 151, Genus Hepatozoon Miller,1908 is characterised by the presence of schizogony cycle in cells- of the internal organs ( liver, spleen, kidney, lung and bone- marrow etc. ) of vertebrate hosts. At an interval of several generations, some merozoites enter: erythrocytes or: leucocytes as in the case may be, and develop into gametocytes. Sporogony stages are knovm to occur in arthropod hosts. Zygote develops into oocyst stage. These increase in size and become ^quite large. Oocysts contain num.erous sporoblasts and ultjjnately to sporozoites. Genus Karyolysus Labbe'', 1894 is distinguished from Haemogregarina and Hepatozoon' on the basis of the following characters. 141 Schizogony occurs in the endothelial cells of the blood vessels and the gametocytes enter the red blood corpuscles of vertebrate hosts., Sporogony occurs in invertebrate host (mite) v/here large oocyst deveops containing sporoblasts. These escape from oocyst as motile vermicules and enter the egg in v/hich these become sporocysts within v/hich sporozoites are-: developed. The above three genera have been reported to occur in the peripheral blood of cold-blooded vertebrates, Gametocytes-of these genera do not always provide a reliable clue to their generic differentiation. As a result, most of the snake haemogregarines have been placed under genus Haemogreqarina although in many cases erythrocytic schizogony - and sporogony of these parasites are unknown. Many species of Hepatozoon have been described from v/arm-blooded vertebrates but there are very- few reports about their pccurrence in cold-blooded animals. In India/ there has-been little work on Hepatozoon. A syrvey of Hepatozoon in ophidians from some parts of West Bengal was undertaken. Results of this survey are given in this chapter. The geographical and hostal differences relating to the haemogregarines described in this work appear to be enough to distinguish them from previously named species given the present ver^^ imperfect state of our knowledge of this group. 1'42 REVIEVI OE LITERATURE Hepatozoon was first seen by Bentley (1905) in leucocytes of dog from rndia,and it was named Leucooytozoon canis by James(1905). Balfour<r905) observed leucocytic parasite of rat,Rattus norveqicus in Sudan and named it.Leucooytozoon muris in 1906, Miller (1908) founded genus Hepatozoon for a parasite of leuco­ cytes of rat and named it Hepatozoon perniciosijm, PTepatozoon in Ophidia. An ophidian Hepatozoon was discovered by Garnham (1950) from an African sv/amp snake, Crotaphopeltis deqeni' which he named Hepatozoon minchini. Hull Sc Camin (1960) described some haem.ogregarines in snalces and stated that these parasites should be placed under genus Hepatozoon,all belonging to a single species, Marguardt (1966) described Hepatozoon sp. in some snaEes in Southern Illinois. Mkrtchyan (1967) described Hepatozoon" ophidia from the blood of Russian snake,Elaphe hohenackeri. Ball, e^t al., (1967) transferred Haemogreqarina rarefaciens to genus Hepatozoon on the basis of thee study of its life cycle,the material being collected from Drymarchon corasis' in Coliraa and Te^ic,Mexico, Clark & Bradford(1969) desc­ ribed five types of a haemoqregaine of the genus Hepatozoon in a single blood smear of a snake,Pituophis melanoleucus mj catenifer collected from Pacific North West. Ball^et al.^ (1969) described Ifepafeozoon fusifex in Boa constrictor collected in Colima ,Mexico. Pessoa S:; Cavalheiro (1969a/ b) and Pessoa.et. CLI ., (1970) described the sporogonic stages of three species of Hepatozoon in water snakes of Brazil. Booden/et al.,(1970) described Hepatozoon sp. in Boa constrictor and its transmission to a lizard by mosquito vector. Oda.et^; al., (1971) observed Hepatozoon in a snake and its transmission. Pessoa Sc 3iasi(1973) described several species of Hepatozoon parasites of Br^silian snakes. Ball S Chao(1973) observed the sporogo- nous stages of Hepatozoon rarefaciens cultured in a Culex pipiens WORIC DCN'fi IN INDIA. Sinha (1980^ described Hepatozoon sp. in a snake/itophiesma stolata collected from West Bengal. Hfe (1981^ also recorded Hepatozoon'sp, Ih a rat snake, Ptyas mucosus collected from Bongaon,24-Parganas,West Bengal. 144 RSrERIALS The following hosts were examined for Hepatozoon Infection in their peripheral blood. -'-• Pt.yQ.s mucosus (Linnaeus) This is commonly known as Dhaman* in Bengal. It belongs to Eanily Colubridae under Order Squamata. It inhabits plains,often in the vicinity of human habitations. It is diurnal,non-poisonous,timid and is capable of climbing trees. Itr is olive-green,brown,yellowish in colour- above with', irregular cross-bars black in colour on posterior half of boSy. Ventral side is yellowish in colour . Posterior ventrals and sub-caudals are edged black. Head is disiilnct from neck. Eyes are large with a round pupil. It is known as rat- eater althbugh it has no particular choice of food. rt is distributed throughout India, Indo-China ,Sri'Lanka and adjoining countries. Five specimens were collected from Bongaon,24-Parganas,West Bengal and two of these were found to harbour Hepatozoon in their peripheral blood. 2» Anphiesma stolata (Linnaeus) This snake is known as striped keel-back. It belongs to Family Colubridae an under Order Squamata. Body is olive-green or brown in colour above t45 with black spots or reticulated cross-bars intersected by- two dorso-lateral yellow stripes. These stripes were well marked on the^hinder part of body. Ventral parts are white sometimes with a black spots on the side of each ventral shield. Head is olive-green in colour and the shields are edged with black. It . : is diurnal^non- poisonous and can be handled easily. It feeds chiefly on frogs and toads. I-tj:. is distributed throughout India, Sri Lanka and adjoining countries. Six speciemens were collected from Chakdah, Nadia/West Bengal and examined. Two of these were positive for Hepatozoon in their peripheral blood, 3. Naja naja (Linnaeus) This poisonous snake is popularly known as'Gdkhura'in Bengal. It belongs to Family Slapidae under Order Sguamata. It is olivaceous or brovTnish to black in colour. Head is not very dis­ tinct from neck. Neck region is dilatable which forms hood. Nostrils are large. Scales are placed obliquely, Eyes are round. It; feeds on rats,mice,toads and frogs. It occurs in Bengal,Eastern Himalaya and in Nepal. 146" Pour specimens were collected from Baruipur,24-Parganas,West Bengal, India,for examination and one was foxind to harbour Hfepatozoon in its peripheral blood, METHODS The following methods were adopted for the prepararation of blopd smears and tissue sections. Blood of snakes were obtained either by clipping the tail region or from the facial vein. Infected snakes were killed and small pieces of tissue from liver and lung were fixed in Bouin's•and Camoy's fixatives. Blood smears were stained in Eeishman's and Giemsa's stains. Tissue sections were stained in Heidenhain "^ iron-haematoxylin and haematoxylin-eosin stains. Measurements of parasites were made from fixed and stained materials and Camera lucida drawings were made, Photomicrographs were also taken in some cases. OB; •^S-iLAT I On 147/ Hepatozoon in Ophidia. parasite No. 1. Hepatozoon n. sp. (a) (FIGS. 1 - 2fe ) OcKurrenoe The parasite was encountered in the peripheral blood smears of rat snakes ( Ptyas mucosus Linnaeua ) collected from Bongaon,24-Parganas/West Bengal,Tndia. Morphology Parasite in peripheral blood were intra-erythro- cytic gametoocytes which were at various stages of develop­ ment. Gametocytes (Pigs.l - 10.) These were abundant in the blood smears. Two distinct morphological types were found depending on their size,shape and staining properties. Sexes of these gametocytes could not however,be conclusively distinguiished. Type. I. ( Figs. 1-4.) Young individuals (Pigs.l - 2 ) were oval or bean-shaped measuring 5.5/um - 9.2/Jm X 1.67um - 3.8/um (average 7 Aim X 3.2/um ). Cytoplasm was homogeneous, non­ granular and stained very faintly . Nucleus was central, non-compact and irregular in shape. It stained deep pink in colour. Average measurement of the nucleus was 3.2/um X 2. 3 Aim • 148 Type. II.. As the development advanced, parasites were enclosed in a polar cap at each end which stained deep blue and fitted over parasite as a cup. Polar caps were very prominent xirith a maximiam length of 2.6 /am. Mature gametocyt^s ( Pigs. 3-4) measured 11/um - 13.2/um X 2.7 AMI - 3.3 Aim ( average 12/um X 3/am ), Cytoplasm was homogeneous, basophilic: and non-granjalar . Nucleus was compact, sub- spherical or oval,central or sub-central v;ith deeply stained chromatins. Average measurement of the nucleus was 3.3 /am X 2.6 /um. Type. rr.. Young individuals (Figs. 5 -6j., were thin and narrow,one end lying broader than other. These measured 9.7 Aim - 11.2 Aim X 1.4/um - 1.6/um ( average 10.5/um X 1.5/um X. Cytoplftsm was homogeneous,non-granu­ lar and faintly stained. Nucleus was elongate or oval sub-central and occupied entire v/idth of cell body. As the development advanced, a capsule developed. Inside the capsule,parasite gradually assumed the shape of a charact­ eristic vermicular form. One end of parasite was sharply bent forming a hook like structure. Other end was broad or tapered slightly. Mature forms ( Pigs. 7 - 10) 4 5 nrr\ T 8 10 11 12 FIGS.I - 10, Camera lucida drawings of Hfepatozoon *H.Sf.
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