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    Characterization of white spot syndrome virus of penaeid shrimp: Genomic cloning and sequencing, structural protein analyzing and sequencing, genetic diversity, pathology and virulence

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    Author
    Wang, Qiong
    Issue Date
    1999
    Keywords
    Biology, Microbiology.
    Agriculture, Animal Pathology.
    Agriculture, Fisheries and Aquaculture.
    Advisor
    Lightner, Donald V.
    
    Metadata
    Show full item record
    Publisher
    The University of Arizona.
    Rights
    Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
    Abstract
    The purpose of this dissertation was to characterize virulence, genomic and protein composition of a newly emerged virus of penaeid shrimp: white spot syndrome virus (WSSV). A partial genomic library, covering approximately 30-50% of the genome, of WSSV isolated from crayfish Orconectes punctimanus, was constructed by digesting viral DNA with endonuclease ClaI and cloning into the system pBluescript-JM109. Three viral inserts of approximately 2.2 kb, 2.8 kb and 6.3 kb, named as QW245, CR44, and QW237 respectively, were sequenced and analyzed. Six geographic isolates of WSSV, from China, India, Thailand, South Carolina, Texas, as well as from crayfish obtained from the US National Zoo in Washington D.C. were compared by electron microscopy (TEM) and SDS-PAGE. All viral isolates contained three major polypeptides of 25, 23 and 19 kDa. A fourth major polypeptide at the 14.5 kDa position was observed in four of the viral isolates. The 19 kDa polypeptide of the crayfish WSSV appeared larger in size than that of the other isolates. Amino acid composition of four of the major structural polypeptides of the South Carolina WSSV was analyzed. The NH₂ terminal amino acids of the 25, 23 and 14.5 kDa polypeptides of the SC WSSV were sequenced as MDLSFTLSVVTA, MEFGNLTNLDVA, and VARGGKTKGRRG, respectively. The genomic composition of the six geographic isolates of WSSV were compared by combining the methods of restriction analysis using nine endonucleases AccI, BglII, ClaI, BamHI, EcoRI, HindII, HaeI, SacI, XhoI and Southern blot hybridization applying three digoxigenin-11-dUTP labeled WSSV genomic probes LN4, C42 and A6. No distinctive difference among five WSSV isolated from penaeid shrimp was detected; differences were observed in the crayfish isolate of WSSV. The virulence of the six geographic isolates of WSSV were compared by per os challenge of Litopenaeus vannamei postlarvae and juveniles, and Farfantepenaeus duorarum juveniles. The Texas WSSV caused higher and more rapid mortalities; the crayfish WSSV caused lower and less rapid mortalities. L. vannamei postlarvae and juveniles were very susceptible to WSSV infection, while Fa. duorarum juveniles showed moderate resistance.
    Type
    text
    Dissertation-Reproduction (electronic)
    Degree Name
    Ph.D.
    Degree Level
    doctoral
    Degree Program
    Graduate College
    Veterinary Science and Microbiology
    Degree Grantor
    University of Arizona
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