[关键词]
[摘要]
卤虫是虾、蟹苗种培育的关键动物性饵料,查清其能否携带白斑综合征病毒,对培育健康苗种至关重要.2002年7月至10月,自盐田中取野生卤虫,经暂养后选20尾抱卵雌虫,每尾置1个烧杯中,投喂单胞藻,产卵后取出亲体;20d后,每个烧杯中取5尾子一代个体,利用巢式体外DNA扩增技术对亲体和子一代成体进行WSSV病毒检测.检测结果显示:卤虫中可扩增出特异性的、与引物设计吻合的DNA片段;DNA测序分析表明,扩增片段的DNA序列与白斑综合征病毒序列一致;野生卤虫携带白斑综合征病毒的阳性率为58%,实验条件下野生卤虫后代的阳性率为43%;同一家系中,亲代与子代携带病毒的情况不完全一致,不能确定白斑综合征病毒可否通过繁殖,在卤虫世代间进行垂直传播.研究结果表明:卤虫很可能是白斑综合征病毒的携带者,苗种培育和养殖期间投喂卤虫可能造成虾、蟹感染白斑综合征病毒,卤虫检疫是培育健康苗种的关键措施.
[Key word]
[Abstract]
Brine shrimp Artemia is the key animal diet during the course of the penaeid shrimp and crabs' larviculture. It is very important for healthy larvae culture to investigate whether it could carry white spot syndrome virus ( WSSV ). In this study, wild Artemia sp. with eggs and experimentally cultured children Artemia were detected using nested-PCR for the presence of WSSV. Wild adult Artemia sp. were sampled from brine pan from July to October, 2002. 20 control groups were ,set up after ten-days' nursery period, that is, one female Artemia with eggs in each 500ml beaker, feeding microalgae. Parents Artemia were taken out after spawning and then frozen at -20℃. About 20 days later, five children ones were taken out and detected by nested-PCR together with the parents, respectively. The results showed that WSSV positive products could be detected in Artemia, not only in wild adult Artemia but also the filial generation, and DNA ,sequence of the amplified product accords with the designed specific amplified one according to sequence analysis using sequence analysis software Bioedit 4.8.8.. The positive rates of wild brine shrimp and the filial generation were 58 % and 43 %, respectively. In addition, there seems to have some relation between parents and filial generation' s detecting results of the same genealogies. However, we could not tell whether WSSV could vertically transmit in Artemia or not, and there still needs some work to further verify this. In conclusion, the results showed that brine shrimp Artemia could possibly act as reservoir or mechanical vector for WSSV, thus, Artemia diet during shrimp or crabs' larviculture might lead to WSSV infection. Therefore, it is necessary to quarantine the brine shrimp before being fed to shrimp or crabs in order to cultivate healthy larvae.
[中图分类号]
[基金项目]
国家高技术研究发展计划(863计划)