本文主要研究内容
作者张宁（2019）在《福建省规模化猪场四种肠道原虫分子流行病学调查和人兽共患风险分析》一文中研究指出：毕氏肠微孢子虫(Enterocytozoon bieneusi)、芽囊原虫(Blastocystis)、隐孢子虫(Cryptosporidium)和蓝氏贾第鞭毛虫(Giardia duodenum)是四种重要的人畜共患肠道原虫,可感染人、野生动物、伴侣动物、鸟类和多种家畜,人类感染常引起不同程度的腹泻,尤其在儿童或免疫缺陷的人群中最为严重。猪是这四种肠道原虫的重要宿主,能够感染多种人兽共患基因型/虫种,为四种肠道原虫感染人类的潜在传染源。为了解四种肠道原虫在福建省猪的感染情况及人兽共患传播风险,本研究于2018年4月至10月采集福建省6个地区规模化猪场共725份猪粪便样本进行四种肠道原虫流行病学特征调查。基于不同位点进行常规PCR/巢式PCR对四种肠道原虫进行检测,经PCR扩增、电泳,测序、序列拼接比对以及系统进化分析,获得如下结果:1.基于毕氏肠微孢子虫核糖体内部转录间隔区(internal transcribed spacer,ITS)的巢式PCR扩增发现毕氏肠微孢子虫总感染率为24.4%(177/725),其中哺乳仔猪感染率最高(36%),公猪的感染率最低(7.1%),经统计学分析,不同群体猪毕氏肠微孢子虫感染率差异显著(P<0.05);不同地区中南平市感染率最高(43.9%)而漳州市最低(8.6%),经统计学分析,不同地区猪毕氏肠微孢子虫感染率呈显著差异(P<0.05);共鉴定出11个已知的毕氏肠微孢子虫基因型(EbpC、EbpA、CHN-RR2、KIN-1、CHG7、CHS5、CM11、CHG23、G、PigEBITS、D)和2个新基因型(FJF和FJS),其中EbpA和EbpC为优势基因型,也为人兽共患基因型,所占阳性比例分别为38.8%(78/201)和32.8%(66/201)。11个已知基因型和2个新基因型全部归属于具有潜在人兽共患风险的Group1中。应用多位点序列分型(multilocus sequence typing,MLST)方法,选择位点MS1、MS3、MS4以及MS7对毕氏肠微孢子虫阳性样本进行多态性和群体遗传结构分析,结果显示177份阳性样品中,共有52份阳性分离株在4个位点同时扩增成功,形成了48个多位点基因型(Multilocus Genotypes,MLGs),强烈的连锁不平衡和有限的基因重组表明猪群毕氏肠微孢子虫为克隆性群体结构。结合该虫在东北地区猪群体的毕氏肠微孢子虫遗传学数据比较分析,发现不同地区猪毕氏肠微孢子虫之间无明显的地理隔离现象。2.基于芽囊原虫小亚基核糖体RNA(Small Subunit ribosome RNA,SSU rRNA)基因位点进行常规PCR扩增发现芽囊原虫总感染率为45%(326/725),其中公猪感染率最高(67.9%),保育猪的感染率最低(12.5%),经统计学分析,不同群体猪的芽囊原虫感染率差异显著(P<0.05);不同地区中漳州市感染率最高(75.3%),福清市最低(32.2%),经统计学分析,不同地区猪毕氏肠微孢子虫感染率呈显著差异(P<0.05)。系统进化分析共鉴定出2种人兽共患基因亚型:ST1和ST5,其中ST5为优势亚型(97.9%,319/326),分布于不同地区和不同群体的猪,ST1占2.1%(7/326),仅在哺乳仔猪(6.9%,2/29)、断奶仔猪(2.8%,1/36)和保育猪(23.1%,3/13)中检测到。3.基于隐孢子虫SSU rRNA基因位点进行巢式PCR扩增发现隐孢子虫总感染率为7.6%(55/725),6个地区感染率为0-12.4%,福清地区感染率最高(12.4%),而漳州地区未检测到隐孢子虫感染,经统计学分析,不同地区隐孢子虫感染率呈显著差异(P<0.05),不同发育阶段的猪群中,育肥猪感染率最高(24.6%)而在公猪中未发现隐孢子虫感染,统计学分析显示不同群体猪隐孢子虫感染率呈显著差异(P<0.05)。本研究共鉴定出2种人兽共患隐孢子虫虫种:C.scrofarum(72.7%,40/55)和C.suis(27.3%,15/55)。C.scrofarum和C.suis感染存在年龄相关性,即在断奶仔猪中C.suis与C.scrofarum感染率接近,而保育猪阶段后则主要以C.scrofarum感染为主。4.通过巢式PCR基于蓝氏贾第鞭毛虫的β-贾第素基因(β-giardin,BG)、谷氨酸脱氢酶基因(glutamate dehydrogenase,GDH)和磷酸丙糖异构酶基因(triose-phosphateisomerase,TPI)的三个位点进行扩增:基于BG位点,检测到195个阳性样本,总感染率为26.9%(195/725),其中三明地区感染率最高为49.2%,莆田地区感染率最低为7.5%,统计学分析表明不同地区感染率为呈显著差异(P<0.05);哺乳仔猪感染率最高(35.1%),育肥猪感染率最低(10.5%),统计学分析表明不同群体猪感染率呈显著差异(P<0.05)。基于BG位点共鉴定出一种人兽共患集聚体类型:集聚体E。基于TPI和GDH基因对195份BG基因位点阳性样本进行多位点分析,分别在TPI和GDH基因位点检测到11个和6个阳性样本,序列分析均为集聚体E,未发现混合感染情况。共6个样本在三个位点同时得到扩增,形成一种MLG。综上所述,本研究利用分子生物学的方法对福建地区四种肠道原虫分子流行病学特征及人兽共患风险进行了调查评估,研究结果为四种肠道原虫在猪群的流行传播提供了参考,为福建省这四种肠道原虫的防控提供了重要的科学数据,研究结果表明需制定综合措施控制猪群中四种肠道原虫的感染,并采取有效手段防止其对人类的传播。
Abstract
bi shi chang wei bao zi chong (Enterocytozoon bieneusi)、ya nang yuan chong (Blastocystis)、yin bao zi chong (Cryptosporidium)he lan shi gu di bian mao chong (Giardia duodenum)shi si chong chong yao de ren chu gong huan chang dao yuan chong ,ke gan ran ren 、ye sheng dong wu 、ban lv dong wu 、diao lei he duo chong jia chu ,ren lei gan ran chang yin qi bu tong cheng du de fu xie ,you ji zai er tong huo mian yi que xian de ren qun zhong zui wei yan chong 。zhu shi zhe si chong chang dao yuan chong de chong yao su zhu ,neng gou gan ran duo chong ren shou gong huan ji yin xing /chong chong ,wei si chong chang dao yuan chong gan ran ren lei de qian zai chuan ran yuan 。wei le jie si chong chang dao yuan chong zai fu jian sheng zhu de gan ran qing kuang ji ren shou gong huan chuan bo feng xian ,ben yan jiu yu 2018nian 4yue zhi 10yue cai ji fu jian sheng 6ge de ou gui mo hua zhu chang gong 725fen zhu fen bian yang ben jin hang si chong chang dao yuan chong liu hang bing xue te zheng diao cha 。ji yu bu tong wei dian jin hang chang gui PCR/chao shi PCRdui si chong chang dao yuan chong jin hang jian ce ,jing PCRkuo zeng 、dian yong ,ce xu 、xu lie pin jie bi dui yi ji ji tong jin hua fen xi ,huo de ru xia jie guo :1.ji yu bi shi chang wei bao zi chong he tang ti nei bu zhuai lu jian ge ou (internal transcribed spacer,ITS)de chao shi PCRkuo zeng fa xian bi shi chang wei bao zi chong zong gan ran lv wei 24.4%(177/725),ji zhong bu ru zai zhu gan ran lv zui gao (36%),gong zhu de gan ran lv zui di (7.1%),jing tong ji xue fen xi ,bu tong qun ti zhu bi shi chang wei bao zi chong gan ran lv cha yi xian zhe (P<0.05);bu tong de ou zhong na ping shi gan ran lv zui gao (43.9%)er zhang zhou shi zui di (8.6%),jing tong ji xue fen xi ,bu tong de ou zhu bi shi chang wei bao zi chong gan ran lv cheng xian zhe cha yi (P<0.05);gong jian ding chu 11ge yi zhi de bi shi chang wei bao zi chong ji yin xing (EbpC、EbpA、CHN-RR2、KIN-1、CHG7、CHS5、CM11、CHG23、G、PigEBITS、D)he 2ge xin ji yin xing (FJFhe FJS),ji zhong EbpAhe EbpCwei you shi ji yin xing ,ye wei ren shou gong huan ji yin xing ,suo zhan yang xing bi li fen bie wei 38.8%(78/201)he 32.8%(66/201)。11ge yi zhi ji yin xing he 2ge xin ji yin xing quan bu gui shu yu ju you qian zai ren shou gong huan feng xian de Group1zhong 。ying yong duo wei dian xu lie fen xing (multilocus sequence typing,MLST)fang fa ,shua ze wei dian MS1、MS3、MS4yi ji MS7dui bi shi chang wei bao zi chong yang xing yang ben jin hang duo tai xing he qun ti wei chuan jie gou fen xi ,jie guo xian shi 177fen yang xing yang pin zhong ,gong you 52fen yang xing fen li zhu zai 4ge wei dian tong shi kuo zeng cheng gong ,xing cheng le 48ge duo wei dian ji yin xing (Multilocus Genotypes,MLGs),jiang lie de lian suo bu ping heng he you xian de ji yin chong zu biao ming zhu qun bi shi chang wei bao zi chong wei ke long xing qun ti jie gou 。jie ge gai chong zai dong bei de ou zhu qun ti de bi shi chang wei bao zi chong wei chuan xue shu ju bi jiao fen xi ,fa xian bu tong de ou zhu bi shi chang wei bao zi chong zhi jian mo ming xian de de li ge li xian xiang 。2.ji yu ya nang yuan chong xiao ya ji he tang ti RNA(Small Subunit ribosome RNA,SSU rRNA)ji yin wei dian jin hang chang gui PCRkuo zeng fa xian ya nang yuan chong zong gan ran lv wei 45%(326/725),ji zhong gong zhu gan ran lv zui gao (67.9%),bao yo zhu de gan ran lv zui di (12.5%),jing tong ji xue fen xi ,bu tong qun ti zhu de ya nang yuan chong gan ran lv cha yi xian zhe (P<0.05);bu tong de ou zhong zhang zhou shi gan ran lv zui gao (75.3%),fu qing shi zui di (32.2%),jing tong ji xue fen xi ,bu tong de ou zhu bi shi chang wei bao zi chong gan ran lv cheng xian zhe cha yi (P<0.05)。ji tong jin hua fen xi gong jian ding chu 2chong ren shou gong huan ji yin ya xing :ST1he ST5,ji zhong ST5wei you shi ya xing (97.9%,319/326),fen bu yu bu tong de ou he bu tong qun ti de zhu ,ST1zhan 2.1%(7/326),jin zai bu ru zai zhu (6.9%,2/29)、duan nai zai zhu (2.8%,1/36)he bao yo zhu (23.1%,3/13)zhong jian ce dao 。3.ji yu yin bao zi chong SSU rRNAji yin wei dian jin hang chao shi PCRkuo zeng fa xian yin bao zi chong zong gan ran lv wei 7.6%(55/725),6ge de ou gan ran lv wei 0-12.4%,fu qing de ou gan ran lv zui gao (12.4%),er zhang zhou de ou wei jian ce dao yin bao zi chong gan ran ,jing tong ji xue fen xi ,bu tong de ou yin bao zi chong gan ran lv cheng xian zhe cha yi (P<0.05),bu tong fa yo jie duan de zhu qun zhong ,yo fei zhu gan ran lv zui gao (24.6%)er zai gong zhu zhong wei fa xian yin bao zi chong gan ran ,tong ji xue fen xi xian shi bu tong qun ti zhu yin bao zi chong gan ran lv cheng xian zhe cha yi (P<0.05)。ben yan jiu gong jian ding chu 2chong ren shou gong huan yin bao zi chong chong chong :C.scrofarum(72.7%,40/55)he C.suis(27.3%,15/55)。C.scrofarumhe C.suisgan ran cun zai nian ling xiang guan xing ,ji zai duan nai zai zhu zhong C.suisyu C.scrofarumgan ran lv jie jin ,er bao yo zhu jie duan hou ze zhu yao yi C.scrofarumgan ran wei zhu 。4.tong guo chao shi PCRji yu lan shi gu di bian mao chong de β-gu di su ji yin (β-giardin,BG)、gu an suan tuo qing mei ji yin (glutamate dehydrogenase,GDH)he lin suan bing tang yi gou mei ji yin (triose-phosphateisomerase,TPI)de san ge wei dian jin hang kuo zeng :ji yu BGwei dian ,jian ce dao 195ge yang xing yang ben ,zong gan ran lv wei 26.9%(195/725),ji zhong san ming de ou gan ran lv zui gao wei 49.2%,pu tian de ou gan ran lv zui di wei 7.5%,tong ji xue fen xi biao ming bu tong de ou gan ran lv wei cheng xian zhe cha yi (P<0.05);bu ru zai zhu gan ran lv zui gao (35.1%),yo fei zhu gan ran lv zui di (10.5%),tong ji xue fen xi biao ming bu tong qun ti zhu gan ran lv cheng xian zhe cha yi (P<0.05)。ji yu BGwei dian gong jian ding chu yi chong ren shou gong huan ji ju ti lei xing :ji ju ti E。ji yu TPIhe GDHji yin dui 195fen BGji yin wei dian yang xing yang ben jin hang duo wei dian fen xi ,fen bie zai TPIhe GDHji yin wei dian jian ce dao 11ge he 6ge yang xing yang ben ,xu lie fen xi jun wei ji ju ti E,wei fa xian hun ge gan ran qing kuang 。gong 6ge yang ben zai san ge wei dian tong shi de dao kuo zeng ,xing cheng yi chong MLG。zeng shang suo shu ,ben yan jiu li yong fen zi sheng wu xue de fang fa dui fu jian de ou si chong chang dao yuan chong fen zi liu hang bing xue te zheng ji ren shou gong huan feng xian jin hang le diao cha ping gu ,yan jiu jie guo wei si chong chang dao yuan chong zai zhu qun de liu hang chuan bo di gong le can kao ,wei fu jian sheng zhe si chong chang dao yuan chong de fang kong di gong le chong yao de ke xue shu ju ,yan jiu jie guo biao ming xu zhi ding zeng ge cuo shi kong zhi zhu qun zhong si chong chang dao yuan chong de gan ran ,bing cai qu you xiao shou duan fang zhi ji dui ren lei de chuan bo 。
论文参考文献
论文详细介绍
论文作者分别是来自福建农林大学的张宁，发表于刊物福建农林大学2019-08-30论文，是一篇关于毕氏肠微孢子虫论文,芽囊原虫论文,隐孢子虫论文,蓝氏贾第鞭毛虫论文,基因分型论文,福建农林大学2019-08-30论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自福建农林大学2019-08-30论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。
标签：毕氏肠微孢子虫论文; 芽囊原虫论文; 隐孢子虫论文; 蓝氏贾第鞭毛虫论文; 基因分型论文; 福建农林大学2019-08-30论文;