云南省独龙牛芽囊原虫分子流行病学调查  

Molecular epidemiological investigation of Blastocystis in Bos frontalis in Yunnan Province

作　　者：庄尔俊 岳凤娇 赵俊杰 李海龙 ZHUANG Erjun;YUE Fengjiao;ZHAO Junjie;LI Hailong(School of Basic Medical Sciences,Dali University,Dali 671000,China)

机构地区：[1]大理大学基础医学院,云南大理671000

出　　处：《黑龙江畜牧兽医》2022年第5期64-67,共4页Heilongjiang Animal Science And veterinary Medicine

基　　金：国家自然科学基金项目(31760726);大理大学媒介生物学创新团队基金项目(ZKLX2019104)。

摘　　要：为了了解云南省怒江州独龙牛芽囊原虫的感染情况及基因型,试验分4个季节从怒江州怒江流域(古泉村、亚左洛村、鸠门当、茨开镇)及独龙江流域(独龙江乡)5个地点采集到987份独龙牛粪便样本,基于芽囊原虫的18S rRNA基因序列设计引物,采用常规PCR方法对粪便DNA进行扩增,琼脂糖凝胶电泳后对阳性样本进行测序分析。使用Clustal X 1.83软件和BLAST程序对测得序列在GenBank上与芽囊原虫参考序列进行同源性比对,通过MEGA-X软件计算序列之间的遗传距离,采用邻接法(NJ)构建系统进化树,以SPSS 19.0软件对感染率进行统计分析。结果表明:共得到119份芽囊原虫阳性样本,与GenBank中单峰驼芽囊原虫SSU rRNA基因序列(KC148207.1)相比,同源性为97.06%~98.92%;与牛芽囊原虫SSU rRNA基因序列(KC148205.1)相比,同源性为96.86%~98.19%,鉴定出ST10、ST14两种基因亚型。独龙牛芽囊原虫的总感染率为12.06%,其中古泉村、亚左洛村、鸠门当、茨开镇、独龙江乡感染率分别为7.28%、12.30%、19.27%、12.28%、9.20%,鸠门当采样点芽囊原虫的感染率与其他采样点比较差异显著(P<0.05)。独龙江流域与怒江流域的芽囊原虫感染率分别为9.20%、12.67%,两者比较差异不显著(P>0.05)。春夏秋冬4个季节芽囊原虫感染率分别为8.03%、11.39%、10.94%、18.22%,春季芽囊原虫感染率最低,冬季最高,两者比较差异显著(P<0.05)。说明怒江州独龙牛存在芽囊原虫感染,不同采样点的芽囊原虫感染率存在差异,冬季芽囊原虫感染率最高,需采取相应措施进行预防。In order to understand the infection and genotype of Blastocystis in Bos frontalis in Nujiang Prefecture, Yunnan Province, the experiment was divided into 4 seasons and collected 987 feces samples of Bos frontalis from 5 locations in Nujiang River Basin(Guquan Village, Yazuoluo Village, Jiumendang, Cikai Town) and Dulong River Basin(Dulongjiang Township) in Nujiang Prefecture. Primers were designed based on the 18 S rRNA gene sequence of Blastocystis;the fecal DNA samples were amplified by conventional PCR method, and the positive samples were sequenced and analyzed after agarose gel electrophoresis. Clustal X 1.83 software and BLAST program were used to carry out homology alignment of the measured sequences with the reference sequence of Blastocystis on GenBank;MEGA-X software was used to calculate the genetic distance between the sequences, and the phylogenetic tree was constructed by Neighbor-Joining method;SPSS 19.0 software was used to carry out statistical analysis of infection rate. The results showed that a total of 119 Blastocystis positive samples were obtained. Compared with the SSU rRNA gene sequence of Camelus dromedarius(KC148207.1) in GenBank, the sequence homology was 97.06%-98.92%, and compared with the SSU rRNA gene sequence of Bos taurus(KC148205.1), the sequence homology was 96.86%-98.19%. Two genotypes, ST10 and ST14, were identified. The total infection rate of Blastocystis in Bos frontalis was 12.06%, among which the infection rates of Guquan Village, Yazoluo Village, Jiumendang, Cikai Town and Dulongjiang Township were 7.28% and 12.30%, 19.27%, 12.28%, 9.20%, respectively. Compared with other sampling sites, the infection rate of Blastocystis in Jiumendang sampling site was significantly different(P<0.05). The infection rates of Blastocystis in the Dulongjiang River basin and Nujiang River basin were 9.20% and 12.67%, respectively;the infection rate in two basins was not significant different(P>0.05). The infection rates of Blastocystis in spring, summer, autumn and winter were 8.03%, 11.3

关 键 词：怒江 独龙牛 芽囊原虫 18S rRNA 分子流行病学 

分 类 号：S823.89[农业科学—畜牧学] S852.729[农业科学—畜牧兽医]