發(fā)布時(shí)間：2018-07-25 10:13

【摘要】：隱孢子蟲(Cryptosporidium spp.)和藍(lán)氏賈第蟲(Giardia lamblia)是兩種重要的機(jī)會(huì)性致病腸道原蟲,已在世界范圍內(nèi)的多個(gè)國家和地區(qū)發(fā)現(xiàn)其感染。Cryptosporidium spp.和G.lamblia宿主譜廣,可感染人和多種動(dòng)物,嚴(yán)重危害人類健康,影響動(dòng)物的生產(chǎn)性能。牛是陜西省重要的經(jīng)濟(jì)動(dòng)物,掌握本省犢牛Cryptosporidium和G.lamblia的感染狀況,明確其種群結(jié)構(gòu),可為其防控提供基礎(chǔ)資料和數(shù)據(jù)。本研究利用巢式PCR技術(shù)分別對(duì)陜西省部分地區(qū)奶牛和秦川牛犢牛的Cryptosporidium和G.lamblia感染狀況進(jìn)行了調(diào)查,并對(duì)其進(jìn)行了種群結(jié)構(gòu)分析,獲得以下結(jié)果:1.對(duì)從陜西省楊凌、咸陽、銅川、漢中、寶雞、渭南6個(gè)地區(qū)采集的371份(198份來自于奶牛,173份來自于秦川牛)犢牛糞便樣品進(jìn)行Cryptosporidium 18S rRNA基因的PCR擴(kuò)增分析發(fā)現(xiàn),Cryptosporidium總感染率為23.18%(86/371),其中奶牛犢牛感染率為31.31%(62/198),秦川牛犢牛感染率為13.87%(24/173),斷奶前犢牛(1~2月齡)感染率為25.00%(50/200),斷奶后犢牛(3~6月齡)感染率為21.05%(36/171)。86份陽性樣品中,36份鑒定為安氏隱孢子蟲(C.andersoni),21份鑒定為瑞氏隱孢子蟲(C.ryanae),29份鑒定為牛隱孢子蟲(C.bovis)。對(duì)36份C.andersoni分離株進(jìn)行基于MS1、MS2、MS3和MS16基因位點(diǎn)的多位點(diǎn)序列分型結(jié)果顯示,有15份樣品在4個(gè)位點(diǎn)均擴(kuò)增成功,分別形成3、1、2、1個(gè)基因亞型,構(gòu)成4個(gè)MLST亞型(A4A4A4A1、A1A4A4A1、A4A4A2A1、A5A4A4A1),全部分布于斷奶后犢牛。2.對(duì)從陜西省楊凌、咸陽、銅川、漢中、寶雞、渭南6個(gè)地區(qū)采集的371份(198份來自于奶牛,173份來自于秦川牛)犢牛糞便樣品進(jìn)行G.lamblia分子檢測(cè),總感染率為18.87%(70/371),其中奶牛感染率為20.71%(41/198),秦川牛感染率為16.76%(29/173),斷奶前犢牛(1~2月齡)感染率為20.00%(40/200),斷奶后犢牛感染率為17.54%(30/171)�；赥PI基因的DNA序列比對(duì)發(fā)現(xiàn)了集聚體E(62/70)和集聚體A(8/70)。集聚體E形成了17個(gè)集聚體亞型(E1~E17),其中E13~E17為本研究中新鑒定的集聚體亞型;集聚體A形成了一個(gè)集聚體亞型A1�；赥PI、BG、GDH和SSU rRNA基因的多位點(diǎn)序列分型結(jié)果顯示,共有37份G.lamblia分離株在4個(gè)位點(diǎn)均擴(kuò)增成功,分別形成9、9、5、2個(gè)集聚體亞型,構(gòu)成25個(gè)不同的MLGs,其中22個(gè)集聚體E的MLGs分布于奶牛和秦川牛的犢牛,1個(gè)集聚體A的MLG分布于奶牛斷奶前犢牛,2個(gè)集聚體A和E混合感染的MLGs,分布于奶牛犢牛。
[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.
【學(xué)位授予單位】：西北農(nóng)林科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：S858.23

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本文編號(hào)：2143488