框镜鲤肠管单极虫病的组织病理学分析及分子鉴定  

Pathohistological analysis and PCR detection of intestinal thelohanellosis in mirror carp(Cyprinus carpio var. specularis)

作　　者：叶彩燕 汪开毓[1,2] 何琦瑶 杨倩[1] 刘韬[1] 余泽辉[1] 谢恒 何晟毓 何洁[1] 赵武义 YE Caiyan;WANG Kaiyu;HE Qiyao;YANG Qian;LIU Tao;YU Zehui;XIE Heng;HE Shengyu;HE Jie;ZHAO Wuyi(Fisheries Department of Sichuan Agricultural University,Chengdu 611130,China;Key Laboratory of Animal Disease and Human Health of Sichuan Province,Sichuan Agricultural University,Chengdu 611130,China)

机构地区：[1]四川农业大学鱼病研究中心,四川成都611130 [2]四川农业大学动物疫病与人类健康四川省重点实验室,四川成都611130

出　　处：《水产学报》2019年第4期1154-1161,共8页Journal of Fisheries of China

基　　金：四川省科技厅应用基础项目(2014JY0143);教育部"长江学者和创新团队发展计划"创新团队项目(IRT0848)~~

摘　　要：2017年6月,四川德阳某养殖场的框镜鲤患病且大量死亡。主要症状为腹部膨大,肠管有大小不等的肿物。为明确其死亡病因,进行了细菌学、寄生虫形态学观察,组织病理学和PCR检测。细菌学检查为阴性;病理组织学观察发现肠道的损伤最为严重,表现为有大量寄生虫样孢囊突出于肠管。寄生虫形态学观察可见孢子长约23～30μm,宽10～15μm,极囊长11～15μm,呈长卵圆形,约占孢子长的1/3～1/2,孢子外面被有一层无色透明的薄膜,前端孢子质中有一个大核和两个小核,与单极虫形态一致;针对吉陶单极虫的18S SSUrRNA基因进行巢式PCR检测,扩增出大小为716 bp的目的片段(命名为TKF-1),目的条带的测序和序列分析结果表明其为吉陶单极虫。根据组织病理特点及PCR检测结果确认框镜鲤的死亡是感染吉陶单极虫所致。In order to identify the pathogen which caused mass mortality in farmed Cyprinus carpio var.specularis in Deyang district of Sichuan Province,China,the pathogen of this event was determined following bacteriology,parasite morphological observation,histopathology and PCR detection.The results showed that bacteriological examination was negative whereas parasite was positive with Thelohanellus kitauei.Histologically,intestinal damage is the most serious,where a large number of parasitoid cysts were noted.Morphological observation showed that the total length and width of the spores were about 23 30μm and 10 15μm,respectively.In addition,the polar capsule was 11 15μm long and ovalellipsoidal,which accounted for about 1/3 1/2 of spores.The outer layer of the spores covered with a thin colorless and transparent film.One pyriform macronucleus was in the upper part and two globular micronuclei near the iodinophilous vacuole.Tissue DNA from infected intestine was identified as T.kitauei(TKF-1)by nest polymerase chain reaction amplification of the 18S SSUrRNA gene and the sequenced 18S SSUrRNA gene of TKF-1 has a homology up to 100%with T.kitauei in GenBank.According to the results we confirmed that the death of C.carpio var.specularis in this region was caused by T.kitauei.

关 键 词：框镜鲤 吉陶单极虫 组织病理 PCR检测 

分 类 号：S941.513[农业科学—水产养殖]