目的为了解青海省犬科动物流行的钩虫种类,为青藏高原地区钩虫的流行分布及种群进化研究提供依据。方法2014—2016年在青海省达日县和兴海县分别收集犬科动物粪便样本,洗脱获取粪便表面动物细胞,通过PCR扩增和测序对粪便标本溯源,以饱...目的为了解青海省犬科动物流行的钩虫种类,为青藏高原地区钩虫的流行分布及种群进化研究提供依据。方法2014—2016年在青海省达日县和兴海县分别收集犬科动物粪便样本,洗脱获取粪便表面动物细胞,通过PCR扩增和测序对粪便标本溯源,以饱和蔗糖水漂浮法进行粪便虫卵检查,收集虫卵提取DNA,以聚合酶链反应-限制性内切酶片段长度多态性(PCR-restriction fragment length polymorphism,PCR-RFLP)法进行虫种鉴定。结果通过动物粪便溯源试验,获得动物线粒体D-loop区域长度为372bp的基因片段,运用基本局部比对搜索工具(basic local alignment search tool,BLAST)分析显示达日县和兴海县动物粪便标本来源分别为家犬和红狐;虫卵内转录间隔区(internal transcribed spacer,ITS)的PCR扩增,获得长度约为544bp的特异性电泳条带,PCR产物经限制性内切酶RsaⅠ和HinfⅠ的酶切反应显示特异性的酶切、电泳条带,两个钩虫样本均被鉴定为锡兰钩虫。结论锡兰钩虫首次在青藏高原地区报道,并在野生红狐体内被发现,有一定的生物学意义。展开更多
Schistosomiasis is a public health concern in Benin. Freshwater snails of the genus Bulinus serve as intermediate hosts for schistosomes, trematode parasites responsible for bilharzia. The urinary form, caused by Schi...Schistosomiasis is a public health concern in Benin. Freshwater snails of the genus Bulinus serve as intermediate hosts for schistosomes, trematode parasites responsible for bilharzia. The urinary form, caused by Schistosoma haematobium, is the most widespread and is transmitted to humans by these mollusks, with Bulinus truncatus and Bulinus globosus being the most important species. Effective strategies to combat the transmission of these parasites require a prior understanding of the molecular characterization of Bulinus snails. For this purpose, 293 Bulinus snails were collected and morphologically identified from two localities in southern Benin, Sô-Ava and Azowlissè. The snails were preserved in absolute alcohol. To achieve the set objectives, DNA was extracted from the collected biological material, and SSU gene fragments were amplified. Using PCR-RFLP, the amplified fragments were digested with the restriction endonucleases HaeIII, HinfI, and DdeI to perform molecular characterization. In this study, 80 individuals of B. globosus and 10 of B. truncatus were subjected to molecular analysis. The PCR-RFLP profiles showed bands of different sizes for the Bulinus species when analyzed with the three endonucleases using the SSU molecular marker. PCR-RFLP analysis revealed that the snails belonged to the freshwater genus Bulinus, including Bulinus globosus and B. truncatus, based on reference profiles from studies conducted in Nigeria, which enabled precise identification of these gastropods. This study provided initial insights, although still incomplete, into the molecular diversity of these species.展开更多
文摘目的为了解青海省犬科动物流行的钩虫种类,为青藏高原地区钩虫的流行分布及种群进化研究提供依据。方法2014—2016年在青海省达日县和兴海县分别收集犬科动物粪便样本,洗脱获取粪便表面动物细胞,通过PCR扩增和测序对粪便标本溯源,以饱和蔗糖水漂浮法进行粪便虫卵检查,收集虫卵提取DNA,以聚合酶链反应-限制性内切酶片段长度多态性(PCR-restriction fragment length polymorphism,PCR-RFLP)法进行虫种鉴定。结果通过动物粪便溯源试验,获得动物线粒体D-loop区域长度为372bp的基因片段,运用基本局部比对搜索工具(basic local alignment search tool,BLAST)分析显示达日县和兴海县动物粪便标本来源分别为家犬和红狐;虫卵内转录间隔区(internal transcribed spacer,ITS)的PCR扩增,获得长度约为544bp的特异性电泳条带,PCR产物经限制性内切酶RsaⅠ和HinfⅠ的酶切反应显示特异性的酶切、电泳条带,两个钩虫样本均被鉴定为锡兰钩虫。结论锡兰钩虫首次在青藏高原地区报道,并在野生红狐体内被发现,有一定的生物学意义。
文摘Schistosomiasis is a public health concern in Benin. Freshwater snails of the genus Bulinus serve as intermediate hosts for schistosomes, trematode parasites responsible for bilharzia. The urinary form, caused by Schistosoma haematobium, is the most widespread and is transmitted to humans by these mollusks, with Bulinus truncatus and Bulinus globosus being the most important species. Effective strategies to combat the transmission of these parasites require a prior understanding of the molecular characterization of Bulinus snails. For this purpose, 293 Bulinus snails were collected and morphologically identified from two localities in southern Benin, Sô-Ava and Azowlissè. The snails were preserved in absolute alcohol. To achieve the set objectives, DNA was extracted from the collected biological material, and SSU gene fragments were amplified. Using PCR-RFLP, the amplified fragments were digested with the restriction endonucleases HaeIII, HinfI, and DdeI to perform molecular characterization. In this study, 80 individuals of B. globosus and 10 of B. truncatus were subjected to molecular analysis. The PCR-RFLP profiles showed bands of different sizes for the Bulinus species when analyzed with the three endonucleases using the SSU molecular marker. PCR-RFLP analysis revealed that the snails belonged to the freshwater genus Bulinus, including Bulinus globosus and B. truncatus, based on reference profiles from studies conducted in Nigeria, which enabled precise identification of these gastropods. This study provided initial insights, although still incomplete, into the molecular diversity of these species.
