Investigation and genetic polymorphism analysis of rodents infected with Echinococcus in Ili Prefecture, Xinjiang Uygur Autonomous Region, China

Bingjie Wang, Li Zhao, Wanli Ban, Xu Zhang, Chenxi Quan, Munila Teliewuhan, Lixiong He, Zhaoyang Chen, Zhuangzhi Zhang
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Abstract

Alveolar echinococcosis (AE) is a life-threatening disease in humans caused by the larval stage of Echinococcus multilocularis. Domestic animals, dogs, foxes, and small mammals constitute the circular chain of AE. To evaluate the infection, distribution, and genetic polymorphism of AE in the Ili Prefecture (Nilka, Xinyuan and Zhaosu), we conducted this survey.In June and July 2018, 267 small mammals were captured using water-infusion and mousetrap methods. Combined pathogenic and molecular biological methods were used to observe the histopathology of Echinococcus carried by rodents, amplify the mitochondrial nad1 gene of the pathogen, and investigate the genotype and haplotype diversity of Echinococcus in rodents in Ili Prefecture.Morphological identification revealed that these captured small mammals belonged to three species, with Microtus gregalis being the dominant species (183/267). Pathological and molecular biological results confirmed that E. multilocularis was the pathogen of echinococcosis in small mammals, with an infection rate of 15.73% (42/267). Among the three areas sampled, the highest infection rate of rodents was 25.45% (14/55) in Nilka County. However, there was no significant difference in the infection rates between regions (χ2 = 5.119, p > 0.05). Of the three captured rodent species, M. gregalis had the highest infection rate of 17.49% (32/183), but there was no significant difference in infection rates between the rodent species (χ2 = 1.364, p > 0.05). Phylogenetic analyses showed that the nad1 gene sequences obtained in this study clustered in the same clade as isolates from China. These isolates contained 21 haplotypes (Hap_1-21); Hap_2 was the most common haplotype (9/42). Furthermore, haplotype diversity (0.925 ± 0.027) and nucleotide diversity (0.01139 ± 0.00119) were higher in the Ili Prefecture than in other regions, indicating that population differentiation was high. Tajima’s D and Fu’s Fs tests were negative (p > 0.10), indicating that the population had expanded. The low fixation index (Fst) ranged from 0.00000 to 0.16945, indicating that the degree of genetic differentiation was different among different populations.In summary, Ili Prefecture is a high incidence area of AE, and Microtus spp. may play an important role in the transmission of AE in this area. The results of this study provide basic data for further study of the molecular epidemiology, genetic differences, and control of E. multilocularis in the Ili Prefecture, Xinjiang.
中国新疆维吾尔自治区伊犁州啮齿动物感染棘球蚴的调查与遗传多态性分析
肺泡棘球蚴病(AE)是由多形棘球蚴幼虫阶段引起的一种威胁人类生命的疾病。家畜、狗、狐狸和小型哺乳动物构成了 AE 的循环链。为评估伊犁州(尼勒克、新源、昭苏)AE的感染、分布和遗传多态性,我们开展了此次调查。2018年6月至7月,我们采用灌水法和捕鼠器法捕获了267只小型哺乳动物。采用病原学和分子生物学相结合的方法,观察了啮齿类动物携带棘球蚴的组织病理变化,扩增了病原的线粒体 nad1 基因,研究了伊犁州啮齿类动物棘球蚴的基因型和单倍型多样性。形态学鉴定结果显示,这些捕获的小型哺乳动物分属 3 个物种,其中灰麂为优势种(183/267)。病理学和分子生物学结果证实,多角体棘球蚴是小型哺乳动物棘球蚴病的病原体,感染率为 15.73%(42/267)。在取样的三个地区中,尼尔卡县的啮齿动物感染率最高,为 25.45%(14/55)。不过,不同地区之间的感染率没有明显差异(χ2 = 5.119,P > 0.05)。在捕获的三种啮齿类动物中,M. gregalis的感染率最高,为17.49%(32/183),但不同啮齿类动物之间的感染率无显著差异(χ2 = 1.364,P > 0.05)。系统进化分析表明,本研究中获得的 nad1 基因序列与中国的分离株属于同一支系。这些分离物包含 21 个单倍型(Hap_1-21);Hap_2 是最常见的单倍型(9/42)。此外,伊犁州的单倍型多样性(0.925 ± 0.027)和核苷酸多样性(0.01139 ± 0.00119)均高于其他地区,表明种群分化程度较高。田岛 D 检验和傅氏 Fs 检验均为阴性(P > 0.10),表明种群已经扩大。综上所述,伊犁州是AE的高发区,小尾寒羊可能在该地区的AE传播中扮演了重要角色。本研究结果为进一步研究多角体圆线虫在新疆伊犁州的分子流行病学、遗传差异和控制提供了基础数据。
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