宁夏部分地区奶牛贾第鞭毛虫分子流行病学调查  

Molecular epidemiological investigation of Giardia intestinalis in dairy cows in parts of Ningxia area

作　　者：黎玉琼 郜军荣 刘芸芸 李红兵 高海慧 赵正伟 于有利 牛晓昊 LI Yuqiong;GAO Junrong;LIU Yunyun;LI Hongbing;GAO Haihui;ZHAO Zhengwei;YU Youli;NIU Xiaohao(Institute of Animal Science,Ningxia Academy of Agriculture and Forestry Sciences,Yinchuan 750002,China;Guyuan City Yuanzhou District Animal Husbandry Technology Promotion Service Center,Guyuan 756000,China;Ningxia Agricultural Reclamation Helan Mountain Dairy Co.,Ltd.,Shizuishan 753000,China)

机构地区：[1]宁夏农林科学院动物科学研究所,银川750002 [2]固原市原州区畜牧技术推广服务中心,宁夏固原756000 [3]宁夏农垦贺兰山奶业有限公司,宁夏石嘴山753000

出　　处：《黑龙江畜牧兽医》2023年第20期83-88,共6页Heilongjiang Animal Science And veterinary Medicine

基　　金：宁夏农业高质量发展和生态保护科技创新示范专项(NGSB-2021-12-06);宁夏回族自治区重点研发计划项目(2022BBF02019);宁夏农林科学院自主创新项目(NKYJ-20-12)。

摘　　要：为了明确宁夏地区奶牛贾第鞭毛虫的优势基因型及其流行特征,试验首先于宁夏地区10个规模化奶牛养殖场采集880份奶牛粪便样品,针对贾第鞭毛虫保守基因——β-贾第素(β-giardin,βg)基因序列设计引物,然后以提取的粪便样品全基因组DNA为模板进行巢式PCR,将PCR产物的测序结果进行BLAST比对分析,鉴定贾第鞭毛虫的基因型,并建立系统进化树,最后对宁夏地区不同城市、不同月龄及腹泻和非腹泻奶牛粪便样品的感染阳性率和基因型进行统计。结果表明:880份粪便样品中,有145份样品扩增到大小为510 bp的条带,贾第鞭毛虫感染阳性率为16.5%(145/880),其中集聚体A的占比为1.4%(2/145),集聚体E的占比为98.6%(143/145);所有集聚体A的βg基因为1种序列(已上传至GenBank,登录号为OQ1012611NX),所有集聚体E的βg基因为4种序列(已上传至GenBank,登录号分别为OQ1012622NX、OQ1012633NX、OQ1012644NX和OQ1012655NX);OQ1012611NX(奶牛)与MK5773339.1(藏牛)、MF497409.1(麝香鹿)和LC437420.1(犬)等在同一分支上;OQ1012622NX(奶牛)、OQ1012633NX(奶牛)、OQ1012644NX(奶牛)与MK862313.1(马)、KR0755939.1(绵羊)、MK610388.1(滩羊)等在同一分支上;OQ1012655NX(奶牛)与KJ188047.1(奶牛)、KJ188056.1(奶牛)、MK5773337.1(藏牛)在同一分支上。宁夏地区四个城市奶牛贾第鞭毛虫感染阳性率表现为中卫市(50.0%,8/16)>银川市(16.3%,126/772)>吴忠市(14.3%,9/63)>石嘴山市(6.9%,2/29);仅在银川市同时检测出集聚体A(1.6%,2/126)和集聚体E(98.4%,124/126),在中卫市、石嘴山市和吴忠市均仅检测出集聚体E;不同月龄奶牛贾第鞭毛虫感染阳性率表现为3~4月龄(26.5%,30/113)>0~2月龄(23.3%,53/227)>5~7月龄(17.4%,49/282)>12月龄以上(6.9%,13/189)>8~12月龄(0,0/69),仅在5~7月龄奶牛粪便中同时检测出集聚体A(4.1%,2/49)和集聚体E(95.9%,47/49),其他月龄段均仅检测出集聚体E;腹泻样品中贾第鞭毛虫感染阳性率为23.6%(37/157),非腹泻In order to clarify the dominant genotype and epidemic characteristics of Giardia intestinalis in dairy cattle in Ningxia area,in this experiment,10 large-scale dairy farms in Ningxia area were selected to collect 880 cow feces samples;primers were designed for the conserved gene sequence of Giardia intestinalisβ-giardin(βg).Nested PCR was performed by using the whole genome DNA of extracted feces samples as a template;the sequencing results of PCR products were analyzed by BLAST to identify the genotype of Giardia intestinalis and establish a phylogenetic tree.Finally,the positive infection rates and genotypes of different cities,different months of age,diarrhea and non-diarrhea dairy cow feces samples in Ningxia area were counted.The results showed that among the 880 feces samples,145 samples were amplified and obtained a band of 510 bp,and the positive rate of Giardia intestinalis was 16.5%(145/880),of which cluster A was 1.4%(2/145)and cluster E was 98.6%(143/145).Theβg gene of all cluster A contained 1 sequence(uploaded to GenBank,accession number OQ1012611NX)and theβg gene of all cluster E contained 4 sequences(uploaded to GenBank,accession numbers OQ1012622NX,OQ1012633NX,OQ1012644NX and OQ1012655NX).OQ1012611NX(cow)was on the same branch with MK5773339.1(Tibetan cattle),MF497409.1(Musk deer)and LC437420.1(dog)and so on;OQ1012622NX(cow),OQ1012633NX(cow),and OQ1012644NX(cow)were on the same branch with MK862313.1(horse),KR0755939.1(sheep),MK610388.1(Beach sheep)and so on;OQ1012655NX(cow)was on the same branch with MK5773337.1(Tibetan cow),KJ188047.1(cow),KJ188056.1(cow).The positive rates of Giardia intestinalis infection in dairy cows in four cities in Ningxia area were as follows:Zhongwei City(50.0%,8/16)>Yinchuan City(16.3%,126/772)>Wuzhong City(14.3%,9/63)>Shizuishan City(6.9%,2/29).Cluster A(1.6%,2/126)and cluster E(98.4%,124/126)were detected simultaneously in Yinchuan City,and only cluster E were detected in Zhongwei City,Shizuishan City and Wuzhong City.The positive rate of Giardia intestinalis i

关 键 词：宁夏地区 贾第鞭毛虫 奶牛 β-贾第素 巢式PCR 粪便样品 

分 类 号：S851.3[农业科学—预防兽医学]