full issue - Association of Biotechnology and Pharmacy
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Current Trends in <strong>Biotechnology</strong> <strong>and</strong> <strong>Pharmacy</strong>Vol. 5 (1) 1011-1020 January 2011. ISSN 0973-8916 (Print), 2230-7303 (Online)1017intranuclear inclusions (Fig. 5a <strong>and</strong> 5b). TheWSSV infection was reported to have occurredfirst in the stomach, gill <strong>and</strong> cuticular epidermis,than subsequently got spread systematically toother t<strong>issue</strong>s <strong>of</strong> mesodermal <strong>and</strong> ectodermal origin(21). The presence <strong>of</strong> dense basophilicintranuclear inclusion <strong>and</strong> chromatin marginationare reported to be characteristic <strong>of</strong> WSSVinfection (19,2). Cytopathological changes in theinfected gut <strong>and</strong> gills clearly suggest WSSVinfection in this shrimps. Our findings are inagreement with the cytopathological changes <strong>of</strong>different t<strong>issue</strong>s <strong>of</strong> WSSV infected shrimpsreported by Chang et al. (10) <strong>and</strong> Yogan<strong>and</strong>hanet al. (22). In the infected muscles also, severenecrosis, chromatin margination <strong>and</strong> densebasophilic intranuclear inclusions (Fig. 5d) havebeen identified indicating the damage caused byviral infection. Histopathological analysis <strong>of</strong>hepatopancreas <strong>of</strong> infected shrimp demonstratedsevere pyknosis <strong>and</strong> karyorrhexis in connectivet<strong>issue</strong> compartment (Fig. 5c). Prominent changeswere not seen in the tubular epithelial cells <strong>of</strong> thehepatopancreas. Our observation is in agreementwith Mohan <strong>and</strong> Shankar (23), who reported thatWSSV does not infect t<strong>issue</strong>s <strong>of</strong> endodermal originsuch as hepatopancreatic tubular epithelium.Nuclear Pyknosis <strong>and</strong> karyorrhexis have beenreported to be associated with several viralinfections (21). Pantoja <strong>and</strong> Lightner (24)observed pyknosis <strong>and</strong> karyorrhexis in animalsexperimentally infected with WSSV. Presentlyobserved Pyknosis <strong>and</strong> karyorrhexis in connectivet<strong>issue</strong> cells <strong>of</strong> hepatopancreas <strong>and</strong> also in gutindicate the role <strong>of</strong> viruses in nuclear damage.The transmission electron microscopyobservations <strong>of</strong> pellet from the filtrate <strong>of</strong> diseasedP. monodon cuticular epidermis stained bynegative staining indicated the presence <strong>of</strong> rodshaped viral particles (Fig. 4). These are similarto the viral particles observed in negative staining<strong>of</strong> spontaneously diseased shrimps reported byChou et al. (8). Electron microscopic observationconformed that the rod shaped virus wasconsidered to be the main causative agent in thepresent study. The virus is reported to be highlypathogenic <strong>and</strong> constitutes a threat to shrimpculture. It is also reported to be transmitted orallyor via water (8). Wongteerasupaya et al. (25)first described the TEM morphology <strong>of</strong> WSSVby negative staining. The presently observedmorphology <strong>of</strong> virus described here from SouthGujarat is very similar to the virus previouslyreported from other parts <strong>of</strong> India (14).In the present study, WSSV infected P.monodon shows differences in the expression<strong>of</strong> proteins in muscle <strong>and</strong> hepatopancreas fromuninfected shrimps. In the infected muscle t<strong>issue</strong>,four new differently expressed proteins withmolecular weight 79kDa, 58kDa, 36kDa <strong>and</strong>17kDa have been detected. In the infectedhepatopancreas, three new proteins withmolecular weight 17kDa, 20kDa <strong>and</strong> 46kDa havebeen detected. A newly expressed protein withmolecular weight 17kDa has been detected inboth the t<strong>issue</strong>s (Fig. 6a <strong>and</strong> 6b). The variation inthe pattern <strong>of</strong> expressions <strong>of</strong> proteins in t<strong>issue</strong>s<strong>of</strong> WSSV infected shrimp is possibly related tothe intensity <strong>of</strong> viral infection, degree <strong>of</strong>pathogenicity <strong>and</strong> t<strong>issue</strong> specific disintegration(14). Van Hulten et al. (26) have reported thatWSSV virion protein ranges from 18kDa to28kDa. Presently observed expression <strong>of</strong> 17kDaprotein in infected muscle <strong>and</strong> hepatopancreasindicates the expression <strong>of</strong> viral protein <strong>and</strong>confirms the presence <strong>of</strong> WSSV infection in boththe t<strong>issue</strong>s. Expression <strong>of</strong> new protein, 17kDa ininfected muscle as well as in hepatopancreas isfound to be similar to the expression <strong>of</strong> newprotein from WSSV infected shrimps reportedby Sathish et al. (27).In terms <strong>of</strong> morphological observations,histopathological effects, viral morphology <strong>and</strong>expression <strong>of</strong> new proteins in infected t<strong>issue</strong>s,Protein expression in WSSV