白斑症病毒(WSSV)对克氏原螯虾血细胞的感染规律  

Infection Process of WSSV in Haemocytes of Procambarus clarkii

作　　者：王轶南[1] 战文斌[1] 唐小千[1] 邢婧[1] 

机构地区：[1]中国海洋大学海水养殖教育部重点实验室,山东青岛266003

出　　处：《中国海洋大学学报（自然科学版）》2010年第9期56-60,共5页Periodical of Ocean University of China

基　　金：国家重点基础研究发展计划项目(2006CB101806);国家高技术研究发展计划项目(2006AA100312)资助

摘　　要：用WSSV粗提液注射感染克氏原螯虾，感染后每24h采集螯虾血细胞。血细胞经与抗WSSV单克隆抗体及FITC标记的羊抗鼠抗体结合反应后，应用流式细胞仪检测WSSV对血细胞的感染，同时记录螯虾累积死亡率及血细胞密度。结果表明，感染WSSV后螯虾1～7d的累积死亡率分别为3．3％，13．3%，16．7％，36．7％，70．0％，90％和100％；1～5d血细胞被感染比例分别为6．47％，6．93％，10．65％，26．08％和4．94％；1～5d被感染细胞的平均荧光强度分别为10．7，10．89，11．71，13．77和15．47；1～6d血细胞密度分别为（7．56±0．30），（5．60±0．24），（4．21±0．30），（1.45±0．26），（1．21±0．21）和（1．14±0．18）×10^6个／mL，阴性对照组螯虾血细胞密度为（5．34±0．22）×10^6个／mL。可见，感染WSSV后螯虾血细胞密度呈现先升后降的趋势，至感染后第6天血细胞密度仅为对照组的21．3％；被感染血细胞内的病毒量始终呈上升趋势、达到最高感染率时螯虾处于濒死状态，表明血细胞的WSSV感染与螯虾死亡密切相关。Procambarus clarkii were injected with WSSV and then haemocytes were collected every 24 h. After reacted with WSSV specific monoclonal antibodies and secondary antibody conjugated with FITC, the haemocytes were detected for WSSV infection using a flow cytometer. Meanwhile, the accumulative mortalities and the haemocyte densities were also recorded. Results showed that after WSSV infection, the accumulative mortalities of Procambarus clarkii were 3. 3%, 13. 3%, 16. 7%, 36. 7% and 70. 0%, respectively, from day 1 to day 7; the infected rates of haemocyte were 6. 47%, 6. 93%, 10. 65%, 26.08% and 4. 94%, respectively, from day 1 to day 5; the mean fluorescence intensities of infected haemocytes were 10. 7, 10. 89, 11.71, 13. 77 and 15. 47, respectively, from day 1 to day 5; the haemocyte densities were （7. 56±0. 30）, （5.60±0. 24）, （4. 21±0. 30）, （1.45±0.26）, （1.21±0. 21） and （1.14±0. 18）× 10^6 cells/mL, respectively, from day 1 to day 6, while the haemocyte density of negative control was （5. 34±0. 22）× 10^6 cells/mL. The results indicated that after Procambarus clarkii infected with WSSV, the density of haemocytes in haemolymph rose at first and then declined, and till day 6 the value was only 21.3% of that in control ;and the WSSV amount in the infected haemocytes kept rising; the infected rate of haemocyte increased to the highest value when Procambarus clarkii went into moribund stage, which revealed that the WSSV infection degree of haemocytes in Procambarus clarkii has a close relationship with its mortality.

关 键 词：克氏原螯虾 血细胞 对虾白斑症病毒 

分 类 号：S917[农业科学—水产科学]