出口白虾种虾病毒性疾病之病理学及分子生物学诊断.doc

出口白蝦種蝦病毒性疾病之病理學及分子生物學診斷Pathological and molecular diagnosis on the viral diseases of exported broodstock shrimp, Litopenaeus vannamei 研究生：莊博智 Chuang, Po-Chih 指導教授：陳瑞雄、蔡信雄 Chern, Rey-Shyong、Tsai, Shinn-Shyong 【摘要】 病毒為蝦病最主要的病原之一，不僅具有高發病率及高死亡率，且會造成蝦類生長遲緩。其中以白點病毒 hite spot syndrome virus；WSSV 、傳染性皮下及造血組織壞死病毒 nfectious hypodermal and haematopoietic necrosis virus；IHHNV 、套拉症病毒 Taura syndrome virus；TSV 和黃頭病病毒 ellow head virus；YHV 常造成全球養蝦產業嚴重的經濟損失，且為出口種蝦必需檢測之疫病。本研究針對此四種病毒進行病理學檢查及分子生物學診斷目前所檢測之檢體中以IHHNV為最多，其比率高達69 18/26 ，其次為WSSV和TSV，各佔15 4/26 ，惟迄今尚未檢出有黃頭病病例。肉眼檢查，感染白點症、傳染性皮下及造血組織壞死及套拉症之白蝦種蝦，皆病變。但在光學顯微鏡下，罹患白點症蝦隻可見嗜鹼性核內包涵體，病變分佈以表皮層出現比例最高 50 ，其次為鰓 25％ 。傳染性皮下及造血組織壞死可見嗜酸性核內包涵體，病變分佈以表皮和輸精管所佔比率最高 63.6％ 。感染套拉症之種蝦，其類淋巴器官為最常被發現病灶之組織 100％ 分子生物學WSSV、IHHNV、TSV和YHV檢測，可各別增幅出941 bp、389 bp、231 bp及135 bp之預期產物。為了達到更快速診斷的目的，本乃針對此四種病毒性疾病建立單管多引子聚合酶鎖反應診斷技術，並證明其可行性。白蝦種蝦、白點症病毒、傳染性皮下及造血組織壞死病毒、套拉症病毒、黃頭病病毒、單管多引子聚合酶連鎖反應 【Abstract】 Viruses are the most major pathogens of shrimp, and they can induce high morbidity, mortality and growth retardation in cultured shrimp. Among them, white spot syndrome virus WSSV , infectious hypodermal and haematopoietic necrosis virus IHHNV , Taura syndrome virus TSV and yellowhead virus YHV often cause heavy economic losses. The present study is to investigate the viral diseases of exported broodstock shrimp by means of pathologic and molecular methods. We collected 26 cases 57 sirmps during 2002 to 2004. The prevalence of IHHNV in white shrimp is more than 69％ 18/26 , followed by WSSV and TSV, each has 15％. However, no YHV case is detected. Grossly, WSSV, IHHNV and TSV have not remarkable lesions. Histopathologic lesion typical for WSSV infection is the presence of basophilic intranuclear inclusion body. The cuticle is the main target tissue with the highest prevalence. IHHNV typically infects the tissues of ectodermal and mesodermal origin, showing eosinophilic intranuclear inclusion bodies. Both cuticle and vas defe