山东嘉祥县牲畜交易市场驴肠道寄生虫调查及驴隐孢子虫多位点序列分型

发布时间：2018-02-28 16:08

  本文关键词： 马属动物 肠道寄生虫 流行病学调查 隐孢子虫 MLST 群体遗传结构　出处：《河南农业大学》2015年硕士论文　论文类型：学位论文

【摘要】：马属动物寄生虫病是现代马业中最常见的一种慢性、隐蔽性疾病,患病动物往往无明显的临床症状,严重时表现为消瘦、贫血、腹泻,甚至是死亡。引起马属动物寄生虫病的病原主要有圆线虫、马副蛔虫、绦虫、球虫、隐孢子虫、贾第虫等,其中隐孢子虫和贾第虫是全球分布的人兽共患原虫病原,可感染多种动物,在世界范围内广泛流行。为了解我国部分地区驴肠道寄生虫感染及流行情况,本研究于2013年3月至2014年5月对养驴相对集中的山东省嘉祥县牲畜交易市场进行驴肠道寄生虫流行病学调查。本次调查共采集驴粪便样品508份,采用卢戈氏碘液染色法和饱和蔗糖漂浮法进行检测,检出寄生虫阳性粪样246份,总感染率为48.43%。共发现8种肠道寄生虫,分别为:球虫、贾第虫、隐孢子虫、圆线虫、蛔虫、绦虫、鞭虫、钩虫。其中以圆线虫感染率最高,为32.48%(165/508),其次是蛔虫,感染率为20.28%(103/508),球虫、贾第虫、隐孢子虫、绦虫、鞭虫、钩虫感染率分别为3.74%(19/508)、0.39%(2/508)、5.91%(30/508)、2.76%(14/508)、0.59%(3/508)、0.20%(1/508)。本次调查发现驴肠道寄生虫混合感染感染率为16.14%(82/508),其中以两种寄生虫混合感染为主,圆线虫和另一种寄生虫混合感染居多。为解析驴源隐孢子虫的种系发育关系及群体遗传结构,本试验选用HSP70、MSC6-7、MSC6-5、CP47、CP56、ML2、Chom3T、RPGR 8个基因位点对本实验室保存的114份驴源隐孢子虫阳性样品进行多位点扩增。首先用GP60基因位点鉴定亚型,成功扩增出69个分离株,包括18个C.parvum(Ⅱd A19G1)分离株,2个Horse genotype分离株和49个Donkey genotype(ⅫA16G1)分离株。经过筛选,CP47和CP56位点扩增效果不好,舍弃这两个位点。共有14个隐孢子虫Donkey genotype样品在所有6个位点(Chom3T、HSP70、ML2、MSC6-5、MSC6-7、RPGR)全部有扩增结果,用多位点序列分型(MLST)方法分析隐孢子虫Donkey genotype群体遗传结构。结果表明,14个隐孢子虫Donkey genotype分离株在上述6个基因位点分别形成2、1、3、4、4、3个亚型且形成7个MLST亚型,连锁不平衡性分析结果表明此次调查的隐孢子虫Donkey genotype为流行性群体结构,暗示驴隐孢子虫病在马属动物群体中的流行是不稳定地的。
[Abstract]:Equine parasitosis is one of the most common chronic and hidden diseases in modern horse industry. The disease animals often have no obvious clinical symptoms. In severe cases, they are characterized by emaciation, anemia and diarrhea. Even death. The main pathogens causing parasitic diseases in equine animals are nematode, ascaris equina, tapeworm, coccidia, Cryptosporidium, giardia, among which Cryptosporidium and Giardia are the worldwide zoonotic protozoan pathogens. In order to understand the infection and epidemic situation of donkey intestinal parasites in some parts of China, From March 2013 to May 2014, an epidemiological survey of donkey intestinal parasites was carried out in the livestock market of Jiaxiang County, Shandong Province, where donkey breeding was relatively concentrated. A total of 508 donkey feces samples were collected in this study. Two hundred and forty-six parasite positive feces were detected by Lugo's iodine solution staining and sucrose floatation. The total infection rate was 48.43. Eight intestinal parasites were found, which were coccidia, giardia, Cryptosporidium, nematode, ascariasis. The infection rate of tapeworm, Trichuris and hookworm was the highest, 32.48% 165 / 508%, followed by Ascaris lumbricoides, and the infection rate was 20.280.103% 508%, coccidia, giardia, Cryptosporidium, tapeworm, Trichuris, The infection rate of hookworm is 3.74 / 508 / 0.390.39 and 5.91 / 5.91 / 3.76 / 5080.59 / 5080.59 / 5080.20 / 5080.20% / 508. this survey found that the infection rate of mixed infection of intestinal parasites in donkey is 16.14% 822% 508%, in which the two parasites are mainly mixed infection. In order to elucidate the phylogenetic relationship and population genetic structure of Cryptosporidium donkeys, nematode and another parasite were most commonly infected. In this experiment, 8 loci of HSP70 / MSC6-7 / CP47 / CP56 / ML2 / Chom3TGR were used to amplify 114 samples of Cryptosporidium donkeys. First, 69 isolates were successfully amplified by using GP60 gene loci to identify the subtype of Cryptosporidium donkeys. There were 18 C.parvum (鈪，

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