长春地区梅花鹿毕氏肠微孢子虫分子流行病学调查

为了掌握长春地区梅花鹿毕氏肠微孢子虫(Enterocytozoon bieneusi)的感染率及基因型种类的分布情况,探究其在人与动物之间传播的源由,本研究于2018年12月-2019年12月期间采集长春地区8个养殖场共计538份梅花鹿粪便样品进行毕氏肠微孢子虫的流行病学调查及基因型分析,经PCR鉴定,测序分析、序列拼接比对以及系统进化分析。试验结果表明,长春地区梅花鹿毕氏肠微孢子虫的感染率为17.84%(96/538),不同养殖场的感染率为0~47.96%;研究共发现14种基因型(BEB6、EbpC、Ⅰ、JLD-Ⅲ、JLD-Ⅸ、JLD-ⅩⅤ、JLD-ⅩⅥ、JLD-ⅩⅦ、JLD-ⅩⅧ、JLD-ⅩⅨ、JLD-ⅩⅩ、JLD-ⅩⅪ、JLD-ⅩⅫ和JLD-ⅩⅩⅢ),其中BEB6为优势基因型。应用多位点序列分型(MLST)方法,选择MS1、MS3、MS4以及MS7位点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,在96份阳性样品中,共有16份阳性分离株在至少2个位点同时扩增成功,形成10个多位点基因型(multilocus genotypes,MLGs),相同的基因型之间存在不同亚型,表明长春地区梅花鹿毕氏肠微孢子虫具有丰富的遗传多样性。本研究应用分子生物学方法对长春地区梅花鹿毕氏肠微孢子虫的流行病情况及基因型进行分析,研究结果不仅为梅花鹿毕氏肠微孢子虫防控提供科学依据,也为防控毕氏微孢子虫在人和动物间的传播提供基础数据,对保障公共卫生与健康意义重大。

Molecular Epidemiological Investigation of Enterocytozoon bieneusi in Sika Deer in Changchun Region

In order to investigate the infection and distribution of genotypes of Enterocytozoon bieneusi in Changchun region,and explore the nature of its transmission between humans and animals.A total of 538 feces samples were collected from 8 farms in Changchun region from December 2018 to December 2019 for epidemiological investigated and genotype analysis.After PCR amplification,electrophoresis identification,sequencing analysis,sequence splicing comparison,and phylogenetic analysis,the following results were obtained:The prevalence rate of Enterocytozoon bieneusi in Sika deer in Changchun region was 17.84%(96/538),and the prevalence in different farms was 0 to 47.96%.There were 14 genotypes of Enterocytozoon bieneusi was found in Sika deer in Changchun area(BEB6,EbpC,Ⅰ,JLD-Ⅲ,JLD-Ⅸ,JLD-ⅩⅤ,JLD-ⅩⅥ,JLD-ⅩⅦ,JLD-ⅩⅧ,JLD-ⅩⅨ,JLD-ⅩⅩ,JLD-ⅩⅪ,JLD-ⅩⅫand JLD-Ⅹ).Moreover,BEB6 was the dominant genotype.The multilocus sequence typing(MLST)method was used to analyze the polymorphism and population genetic structure of Enterocytozoon bieneusi positive samples by selecting gene loci MS1,MS3,MS4,and MS7.The results showed that 16 positive isolates of 96 positive samples were successfully amplified at two or more loci simultaneously,forming 10 multilocus genotypes(MLGs).At the same time,there were different subtypes between the same genotype.This result indicated that there was a rich genetic diversity of Enterocytozoon bieneusi of Sika deer in Changchun.In this study,molecular biological methods were used to analyze the epidemic situation and genotypes of Enterocytozoon bieneusi of Sika deer in Changchun area.The results not only provided a scientific basis for the prevention and control of Enterocytozoon bieneusi in Sika deer,but also provided basic data for the prevention and control of the spread of Enterocytozoon bieneusi between humans and animals,which was of great significance to public health and health.