陕西部分地区犊牛隐孢子虫和蓝氏贾第虫种群结构研究
发布时间：2018-07-25 10:13
【摘要】：隐孢子虫(Cryptosporidium spp.)和蓝氏贾第虫(Giardia lamblia)是两种重要的机会性致病肠道原虫,已在世界范围内的多个国家和地区发现其感染。Cryptosporidium spp.和G.lamblia宿主谱广,可感染人和多种动物,严重危害人类健康,影响动物的生产性能。牛是陕西省重要的经济动物,掌握本省犊牛Cryptosporidium和G.lamblia的感染状况,明确其种群结构,可为其防控提供基础资料和数据。本研究利用巢式PCR技术分别对陕西省部分地区奶牛和秦川牛犊牛的Cryptosporidium和G.lamblia感染状况进行了调查,并对其进行了种群结构分析,获得以下结果:1.对从陕西省杨凌、咸阳、铜川、汉中、宝鸡、渭南6个地区采集的371份(198份来自于奶牛,173份来自于秦川牛)犊牛粪便样品进行Cryptosporidium 18S rRNA基因的PCR扩增分析发现,Cryptosporidium总感染率为23.18%(86/371),其中奶牛犊牛感染率为31.31%(62/198),秦川牛犊牛感染率为13.87%(24/173),断奶前犊牛(1~2月龄)感染率为25.00%(50/200),断奶后犊牛(3~6月龄)感染率为21.05%(36/171)。86份阳性样品中,36份鉴定为安氏隐孢子虫(C.andersoni),21份鉴定为瑞氏隐孢子虫(C.ryanae),29份鉴定为牛隐孢子虫(C.bovis)。对36份C.andersoni分离株进行基于MS1、MS2、MS3和MS16基因位点的多位点序列分型结果显示,有15份样品在4个位点均扩增成功,分别形成3、1、2、1个基因亚型,构成4个MLST亚型(A4A4A4A1、A1A4A4A1、A4A4A2A1、A5A4A4A1),全部分布于断奶后犊牛。2.对从陕西省杨凌、咸阳、铜川、汉中、宝鸡、渭南6个地区采集的371份(198份来自于奶牛,173份来自于秦川牛)犊牛粪便样品进行G.lamblia分子检测,总感染率为18.87%(70/371),其中奶牛感染率为20.71%(41/198),秦川牛感染率为16.76%(29/173),断奶前犊牛(1~2月龄)感染率为20.00%(40/200),断奶后犊牛感染率为17.54%(30/171)。基于TPI基因的DNA序列比对发现了集聚体E(62/70)和集聚体A(8/70)。集聚体E形成了17个集聚体亚型(E1~E17),其中E13~E17为本研究中新鉴定的集聚体亚型;集聚体A形成了一个集聚体亚型A1。基于TPI、BG、GDH和SSU rRNA基因的多位点序列分型结果显示,共有37份G.lamblia分离株在4个位点均扩增成功,分别形成9、9、5、2个集聚体亚型,构成25个不同的MLGs,其中22个集聚体E的MLGs分布于奶牛和秦川牛的犊牛,1个集聚体A的MLG分布于奶牛断奶前犊牛,2个集聚体A和E混合感染的MLGs,分布于奶牛犊牛。
[Abstract]:Cryptosporidium (Cryptosporidium spp.) (Giardia lamblia) and Giardia lamblia are two important opportunistic intestinal protozoa, which have been found in many countries and regions all over the world. And G.lamblia host spectrum, can infect people and many animals, serious harm to human health, affect the performance of animals. Cattle are important economic animals in Shaanxi Province. It can provide basic data and data for prevention and control of calf Cryptosporidium and G.lamblia infection in Shanxi province. In this study, nested PCR technique was used to investigate the Cryptosporidium and G.lamblia infection of dairy cows and Qinchuan calves in some areas of Shaanxi Province, and the population structure was analyzed. The following results were obtained: 1. From Shaanxi Province Yang Ling, Xianyang, Tongchuan, Hanzhong, Baoji, The total infection rate of Cryptosporidium 18s rRNA gene was 23.18% (86 / 371) in fecal samples of 371 calves (198 samples from cows and 173 samples from Qinchuan cattle) collected from 6 regions of Weinan, including 31.31% (62 / 198) of cow calves and 31.31% (62 / 198) of calves from Qinchuan calves, and the total infection rate of Cryptosporidium was 23.18% (86 / 371). The infection rate was 13.87% (24 / 173) in cattle, 25.00% (50 / 200) in preweaning calves (1 ~ 2 months old) and 21.05% (36 / 171) .86 positive samples of postweaning calves (36 / 171) were identified as Cryptosporidium Angiosporidium (C.andersoni) 21 were identified as C.ryanae and 29 were identified as C.bovis. The results of multilocus typing based on MS1MS-2MS3 and MS16 gene loci of 36 C.andersoni isolates showed that 15 samples were successfully amplified at 4 loci, forming 3A1m2and 1 gene subtype, respectively, forming four MLST subtypes (A4A4A4A1A1A1A1A4A2A1A1A4A4A1), all of which were distributed in postweaning calves (A4A4A4A2A2A1A4A4A4A1). The feces samples from Yang Ling, Xianyang, Tongchuan, Hanzhong, Baoji and Weinan of Shaanxi Province were detected by G.lamblia molecular method. The total infection rate was 18.87% (70 / 371), including 20.71% (41 / 198) for cows, 16.76% (29 / 173) for Qinchuan cattle, 20.00% (40 / 200) for calves before weaning and 17.54% (30 / 171) for calves after weaning. The DNA sequence alignment based on TPI gene found the agglomeration E (62 / 70) and agglomeration A (8 / 70). Agglomeration E formed 17 agglomeration subtypes (E1~E17), among which E13~E17 was the newly identified agglomeration subtype and agglomeration A formed one agglomeration subtype A1. The results of multilocus sequence typing based on GDH and SSU rRNA gene showed that 37 G.lamblia isolates were successfully amplified at 4 loci, forming 9 GDH subtypes and 2 agglomeration subtypes, respectively. Among the 25 different MLGs, 22 MLGs of agglomeration E were distributed in calves of cows and Qinchuan cattle, 1 MLG of agglomeration A was distributed in pre-weaning calves, and 2 agglomeration A and E co-infected MLGs were distributed in dairy calves.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S858.23
本文编号：2143488
[Abstract]:Cryptosporidium (Cryptosporidium spp.) (Giardia lamblia) and Giardia lamblia are two important opportunistic intestinal protozoa, which have been found in many countries and regions all over the world. And G.lamblia host spectrum, can infect people and many animals, serious harm to human health, affect the performance of animals. Cattle are important economic animals in Shaanxi Province. It can provide basic data and data for prevention and control of calf Cryptosporidium and G.lamblia infection in Shanxi province. In this study, nested PCR technique was used to investigate the Cryptosporidium and G.lamblia infection of dairy cows and Qinchuan calves in some areas of Shaanxi Province, and the population structure was analyzed. The following results were obtained: 1. From Shaanxi Province Yang Ling, Xianyang, Tongchuan, Hanzhong, Baoji, The total infection rate of Cryptosporidium 18s rRNA gene was 23.18% (86 / 371) in fecal samples of 371 calves (198 samples from cows and 173 samples from Qinchuan cattle) collected from 6 regions of Weinan, including 31.31% (62 / 198) of cow calves and 31.31% (62 / 198) of calves from Qinchuan calves, and the total infection rate of Cryptosporidium was 23.18% (86 / 371). The infection rate was 13.87% (24 / 173) in cattle, 25.00% (50 / 200) in preweaning calves (1 ~ 2 months old) and 21.05% (36 / 171) .86 positive samples of postweaning calves (36 / 171) were identified as Cryptosporidium Angiosporidium (C.andersoni) 21 were identified as C.ryanae and 29 were identified as C.bovis. The results of multilocus typing based on MS1MS-2MS3 and MS16 gene loci of 36 C.andersoni isolates showed that 15 samples were successfully amplified at 4 loci, forming 3A1m2and 1 gene subtype, respectively, forming four MLST subtypes (A4A4A4A1A1A1A1A4A2A1A1A4A4A1), all of which were distributed in postweaning calves (A4A4A4A2A2A1A4A4A4A1). The feces samples from Yang Ling, Xianyang, Tongchuan, Hanzhong, Baoji and Weinan of Shaanxi Province were detected by G.lamblia molecular method. The total infection rate was 18.87% (70 / 371), including 20.71% (41 / 198) for cows, 16.76% (29 / 173) for Qinchuan cattle, 20.00% (40 / 200) for calves before weaning and 17.54% (30 / 171) for calves after weaning. The DNA sequence alignment based on TPI gene found the agglomeration E (62 / 70) and agglomeration A (8 / 70). Agglomeration E formed 17 agglomeration subtypes (E1~E17), among which E13~E17 was the newly identified agglomeration subtype and agglomeration A formed one agglomeration subtype A1. The results of multilocus sequence typing based on GDH and SSU rRNA gene showed that 37 G.lamblia isolates were successfully amplified at 4 loci, forming 9 GDH subtypes and 2 agglomeration subtypes, respectively. Among the 25 different MLGs, 22 MLGs of agglomeration E were distributed in calves of cows and Qinchuan cattle, 1 MLG of agglomeration A was distributed in pre-weaning calves, and 2 agglomeration A and E co-infected MLGs were distributed in dairy calves.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S858.23
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