CAJ | 학술논문

锯缘青蟹(Scylla serrata)俗称青蟹,是我国重要的海水养殖蟹类.近年来,浙江、福建、广东等青蟹主要养殖地区出现了严重的青蟹病害.对浙江省养殖青蟹的发病原因和流行病学调查发现,白斑综合征病毒(white spotsyndrome virus,WSSV)与青蟹发病存在较大相关性.为进一步研究WSSV对青蟹的致病性和发病机理,作者采用白斑综合征病毒的除菌过滤液,以1:10-1:10000稀释度注射感染青蟹,结果表明1:10、1:100感染组的青蟹死亡率达100%,1:1000感染组死亡率为66.7%,1:10000感染组死亡率为38.9%.根据攻毒悬液的病毒浓度计算出WSSV对青蟹的LD_(50)为1.19×10~6拷贝/只(7.93×10~3拷贝/g组织);取WSSV感染青蟹血淋巴进行PCR检测,攻毒死亡青蟹的WSSV检出率为100%,表明WSSV对青蟹有很强的致病力.分析病毒感染濒死蟹的血清酚氧化酶(PO)、过氧化物酶(POD)、超氧化物歧化酶(SOD)、碱性磷酸酶(ALP)、谷丙转氨酶(GPT)、符草转氨酶(GOT)等主要酶指标,发现病毒感染青蟹的PO、POD和SOD活力明显低于对照组,而ALP、GPT和GOT的活力则明显高于对照组;用WSSV单克隆抗体对感染蟹进行免疫组化分析,发现WSSV主要侵染青蟹的鳃、甲壳下表皮、心脏、肠、胃等组织的上皮细胞,尤其以鳃上皮细胞损害最为严重.
거연청해(Scylla serrata)속칭청해,시아국중요적해수양식해류.근년래,절강、복건、엄동등청해주요양식지구출현료엄중적청해병해.대절강성양식청해적발병원인화류행병학조사발현,백반종합정병독(white spotsyndrome virus,WSSV)여청해발병존재교대상관성.위진일보연구WSSV대청해적치병성화발병궤리,작자채용백반종합정병독적제균과려액,이1:10-1:10000희석도주사감염청해,결과표명1:10、1:100감염조적청해사망솔체100%,1:1000감염조사망솔위66.7%,1:10000감염조사망솔위38.9%.근거공독현액적병독농도계산출WSSV대청해적LD_(50)위1.19×10~6고패/지(7.93×10~3고패/g조직);취WSSV감염청해혈림파진행PCR검측,공독사망청해적WSSV검출솔위100%,표명WSSV대청해유흔강적치병력.분석병독감염빈사해적혈청분양화매(PO)、과양화물매(POD)、초양화물기화매(SOD)、감성린산매(ALP)、곡병전안매(GPT)、부초전안매(GOT)등주요매지표,발현병독감염청해적PO、POD화SOD활력명현저우대조조,이ALP、GPT화GOT적활력칙명현고우대조조;용WSSV단극륭항체대감염해진행면역조화분석,발현WSSV주요침염청해적새、갑각하표피、심장、장、위등조직적상피세포,우기이새상피세포손해최위엄중.
Mud crab, Scylla serrata (Forskal) , is main economically cultivated marine crab in China, with production of 108,500 tons, 88.14% of the total production of the world. The main mud crab region distributed along the coastline of Southeast China, including Zhejiang, Fujian, Guangdong, Guangxi and Hainan Province. Recently, serious diseases occurred in main mud crab culture region, with the morbidity of 13.77% in cultivated mud crab in 2006 in whole China, resulting in 32.71 million dollars lost. White spot syndrome virus (WSSV) , the most pathogen of shrimp culture in China, was considered an important pathogen in cultivated mud crab. It showed that WSSV detection with PCR was closely relative with occurring of mud crabs diseases during the epidemics investigations from 2006 to 2007. To further study the pathoge-nicity of WSSV to mud crab, WSSV bacteria-free suspension, collected from typical WSSV infected black tiger shrimp (Penaeus monodon) , was used for challenging to healthy mud crab by intramuscular injection at 10-folded dilutions ranged from 1:10 to 1:10000. The results showed that the cumulative mortalities for dilutions of 1:10 to 1:100, 1:1000, 1: 10000 were 100% , 66.7% and 38.9% respectively. The LD50 of WSSV to mud crab was calculated as 1.19×10~6 copies/crab, or 7.93×10~3 copies/g of crab weight by Muench-Reed methods. Gills and hemolymph of moribund mud crabs challenged with WSSV were positive by PCR. These results indicated that WSSV was highly virulent to mud crab. The activities of main hemolymph enzymes, including phenoloxidase ( PO ) , peroxidase ( POD ) , superoxide dis-mutase (SOD) , alkaline phosphatase (ALP) , glutamate-pyruvate transaminase (GPT) and glutamic-oxaloacetic transam-inase (GOT) of mud crab were analyzed. WSSV infected moribund crabs showed significantly descent in activities of PO, POD and SOD, while ascent in ALP, GPT and GOT compared with the control group. WSSV mainly distributed in epithe-lia of gills, cuticular of skeletal plate, heart, intestine and stomach by immunohistochemistry with Mabs against WSSV, and gill epithelia showed heavily damaged with the main distribution of WSSV in nucleic. It can be concluded that WSSV was a main pathogen of mud crab with high pathogenicity.