我国北方海洋经济动物病害纤毛虫的快速检测研究

Pathogenic ciliates are a group of parasites commonly found in marine economic animals. The diseases caused by these pathogens have been emerging problems inflicting significant economic loss in aquaculture in northern China. Currently, no effective antidote is available yet for the control and mitigation of these diseases, which makes the prompt detection of a possible infection be a critical early warning approach to prevent serious losses. The early detection based on traditional morphological methods, however, has been hampered by the small size of pathogenic ciliates and their morphological similarity to closely related species. Therefore, it is urgent in establishing a new identification system for the rapid detection and early warning. In this proposed study, by using the modern taxonomic methods in combination with phylogenetic analyses of the sequences of multiple genes, we aim to (i) make available a comprehensive identification key with photographs for marine pathogenic ciliates in northern China; (ii) establish systems and DNA probe kits for the rapid detection of pathogenic ciliates based on fluorescence in situ hybridization (FISH) targeted species-specific oligonucleotide probes, in combination with the identification key with photographs; and (iii) clarify confusions in their taxonomy and phylogeny. The accomplishment of this research is believed to improve our knowledge about the diversity and distribution of these organisms in the northern China, and provide protocols and kits for fast detections of the pathogenic ciliates, with the aim of reducing economic losses caused by these pathogens.

病害纤毛虫是海洋经济动物中常见且危害严重的原生动物病害原之一，已给我国北方水产养殖业造成巨大经济损失并呈加剧趋势。然而，对于纤毛虫病仍无有效的治疗手段，而且基于传统分类鉴定方法的病原检测难以满足生产中预防预警的需要。因此，构建快速、准确的分类鉴定与检测体系，进行预防预警就成为一项亟需的工作。本研究拟针对我国北方海洋经济动物病害纤毛虫开展采样调查，采用纤毛虫现代分类学研究技术，建立基于形态的图解式分类检索；进行多基因测序分析，针对目标类群特异性基因片段设计DNA探针试剂盒，结合荧光原位杂交（FISH）技术进行分子鉴定，从而构建基于特异性DNA探针、FISH技术结合形态分类图解的快速检测体系。同时，通过形态学结合多基因序列分析，澄清分类学误识，厘清系统地位混乱或不明类群的系统发育关系。本研究将在完善我国北方海域病害纤毛虫分类构成和流行情况的基础上，为该类病害的预防预警提供快速有效的检测手段。

病害纤毛虫是海洋经济动物中常见且危害严重的原生动物病害原之一，已给我国北方水产养殖业造成巨大经济损失并呈加剧趋势。然而，对于纤毛虫病仍无有效的治疗手段，而且基于传统分类鉴定方法的病原检测难以满足生产中预防预警的需要。因此，构建快速、准确的分类鉴定与检测体系，进行预防预警就成为一项亟需的工作。本研究针对我国北方海洋经济动物病害纤毛虫开展采样调查，采用纤毛虫现代分类学研究和原位杂交分子检测技术，针对9种较难鉴定的海洋病害纤毛虫开发了8个快速检测试剂盒，针对10种常见海洋病害纤毛虫构建了可快速鉴定的图解式检索表。同时，通过形态学结合多基因序列分析，澄清缘毛类纤毛虫的单系性和伪角虫属内部亲缘关系问题。本研究在完善我国北方海域病害纤毛虫分类构成和流行情况的基础上，为该类病害的预防预警提供快速有效的检测手段。