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)Protein Expression <strong>and</strong> Microscopic Evidence <strong>of</strong> White SpotSyndrome Virus (WSSV) in Tiger prawns Penaeus monodonBhatt, S.S. a* , Chavda, D. V. a <strong>and</strong> Sreepada, R. A. baB.R. Doshi School <strong>of</strong> Biosciences, Zoology Laboratory, Bakrol Vadtal Road,Satellite Campus, Sardar Patel University, Vallabh Vidhyanagar 388120, IndiabNational Institute <strong>of</strong> Oceanography, Dona Paula, Goa 403 004, India*For Correspondence - bhatt.sujata@gmail.com1011AbstractShrimp culture is being practiced in largeareas <strong>of</strong> coastal farms <strong>of</strong> South Gujarat region inWestern India. The disease caused due to whitespot syndrome virus (WSSV) has been detectedin the cultured Penaeus monodon in this region.White spots were clearly visible in the carapace<strong>of</strong> infected shrimps. The scanning electronmicrographs <strong>of</strong> the upper <strong>and</strong> lower surfaces <strong>of</strong>carapace revealed the presence <strong>of</strong> white spotsmeasuring about 78µm to 135µm. In the infectedshrimps, rod shaped viral particles have also beenidentified from the epidermis associated withcarapace by TEM using negative staining. Thehisto-pathological analysis <strong>of</strong> gills,hepatopancreas, gut <strong>and</strong> muscles <strong>of</strong> P. monodoninfected with WSSV indicated hypertrophiednuclei, presence <strong>of</strong> intranuclear inclusion bodies<strong>and</strong> chromatin margination in gill epithelial cells<strong>and</strong> necrosis <strong>of</strong> gill lamellae. In the infectedmuscles, necrosis, haemocytic infiltration <strong>and</strong>degeneration <strong>of</strong> muscle fibers were clearlyevident. Damage to the mucosal epithelial cells<strong>of</strong> gut was accompanied by haemocyticinflammatory response. Inflammatory response<strong>and</strong> necrotic changes in hepatopancreas <strong>of</strong>infected shrimps have been observed.Hypertrophied <strong>and</strong> piknotic nuclei as well asinfiltration <strong>of</strong> viral particles into connective t<strong>issue</strong>sassociated with vacuolization were observed inall the analyzed infected t<strong>issue</strong>s. Changes in theexpression <strong>of</strong> proteins have been detected bySDS-PAGE in the infected muscles <strong>and</strong>hepatopancreas. Four new proteins <strong>of</strong> 79kDa,58kDa, 36kDa <strong>and</strong> 17kDa were found to bedifferentially expressed in muscles. Inhepatopancreas, three proteins viz. 46kDa, 20kDa<strong>and</strong> 17kDa were differentially expressed ininfected shrimps; whereas 62kDa protein, whichwas expressed in healthy shrimps, was notexpressed in infected shrimps. No change in theprotein expression pattern was detected in gills.The current study has shown that WSSV, besidescausing damage to the t<strong>issue</strong>s, has also drasticallyaltered the protein expression patterns <strong>of</strong> theinfected t<strong>issue</strong>s <strong>of</strong> P. monodon.Key words: WSSV, protein expression, Penaeusmonodon, SDS-PAGEIntroductionIntensive shrimp aquaculture is one <strong>of</strong>the most important aquaculture practices in theworld. Intensive culture practice has also resultedin the development <strong>of</strong> diseases including viraldiseases in shrimp culture units (1). White spotsyndrome virus (WSSV) is one <strong>of</strong> the highlypathogenic shrimp viruses affecting the shrimpculture worldwide with severe economic losses;<strong>and</strong> its characterization is critical for developingeffective control methods (2,3). Though, prawnsinfected with WSSV or YHV have been reportedto be safe for human consumption (4); theProtein expression in WSSV