新疆阿克苏地区羊泰勒虫PCR检测及遗传多样性分析

doi:10.16431/j.cnki.1671-7236.2025.03.031

摘要/Abstract

摘要： 【目的】探究新疆阿克苏地区柯坪县和阿瓦提县羊泰勒虫分类和遗传多样性及发病情况。【方法】通过PCR方法对柯坪县和阿瓦提县养殖场的78份绵羊血液样品进行羊泰勒虫病病原检测，将阳性样本进行测序，将获得的序列与GenBank数据库中泰勒虫种18S rRNA序列进行相似性对及遗传多样性分析。【结果】柯坪县和阿瓦提县羊泰勒虫病的PCR总阳性率为84.6%(66/78)，柯坪县阳性率为73.3%(22/30)，阿瓦提县阳性率为91.7%(44/48)。测序获得的6条代表性序列相似性＞93%，与GenBank数据库中绵羊泰勒虫18S rRNA相似性＞97%。遗传进化树显示，绵羊泰勒虫、环形泰勒虫、鹿泰勒虫等9种泰勒虫具有独立分支，绵羊泰勒虫与环形泰勒虫为近缘种，与土耳其、伊拉克和沙特阿拉伯分离株亲缘关系较近。对中国、伊拉克、土耳其、沙特阿拉伯等7个国家共50条绵羊泰勒虫18S rRNA基因序列进行单倍型分析后发现，这些序列分布于16个单倍型中。本研究所获得的其中5条序列与来自中国河南、甘肃的序列分布在Hap_1，另1条作为Hap_16单独存在。以中国、伊拉克、土耳其、沙特阿拉伯4个国家共47条序列对绵羊泰勒虫种群结构进行分析，结果表明4个国家的绵羊泰勒株均存在分离位点，伊拉克分离株核苷酸多样性高，中国、土耳其、沙特阿拉伯分离株核苷酸多样性低。土耳其分离株平均差异数值最高，说明种群内基因频率分布分散，遗传差异大；中国、伊拉克、沙特阿拉伯分离株平均差异数值较低，说明遗传差异较小。土耳其、伊拉克、沙特阿拉伯分离株的单倍型多样性＞0.5，以伊拉克分离株值最高，反映了较高的遗传多样性，中国分离株单倍型多样性为0.250，说明遗传多样性较低。上述4个国家的Tajima’s D值均＜0，说明种群偏移了遗传平衡状态，可能是种群扩张的结果。【结论】新疆阿克苏地区柯坪县和阿瓦提县的羊普遍存在泰勒虫感染的情况，其主要病原为绵羊泰勒虫，未检测到尤氏泰勒虫与吕氏泰勒虫，绵羊泰勒虫与这2种泰勒虫亲缘关系较远；来自不同地区的绵羊泰勒虫种群单倍型和遗传多样性丰富，且种群正在发生扩张。本研究结果可为新疆阿克苏地区羊泰勒虫病的综合防控及地方养殖业的健康发展提供参考。

关键词: 羊; 泰勒虫; PCR检测; 遗传进化; 防控

Abstract: 【Objective】 The purpose of this experiment was to investigate the classification,genetic diversity and incidence of Theileria of sheep in Keping and Aatii counties,Aksu region,Xinjiang.【Method】 A total of 78 sheep blood samples from farms in Keping and Awati counties were tested for the pathogen of ovine theileriosis by PCR,and the positive samples were sequenced,and the sequences obtained were analyzed for similarity and genetic diversity with 18S rRNA sequences of Theileria species in GenBank.【Result】 The overall PCR positive rate of ovine theileriosis was 84.6% (66/78) in the two counties,with rates of 73.3% (22/30) in Keping county and 91.7% (44/48) in Awati county.The similarity of 6 representative sequences obtained by sequencing was more than 93%,and their similarity with 18S rRNA sequences of Theileria ovis in GenBank database was more than 97%.The genetic evolution tree revealed independent clades for 9 species of Theileria ovis,Theileria annulata and Theileria cervi,and so on.Theileria ovis was closely related to Theileria annulata,and was closely related to isolates from Turkey,Iraq and Saudi Arabia.Haplotype analysis of 50 18S rRNA gene sequences from seven countries including China,Iraq,Turkey,Saudi Arabia showed that these sequences were distributed in 16 haplotypes.Five sequences from this study were grouped into Hap_1 along with sequences from Henan and Gansu provinces in China,and the other one was isolated as Hap_16.Population structure analysis using a total of 47 sequences from China,Iraq,Turkey and Saudi Arabia indicated a separation site in all four countries.Strains from Iraq exhibited high nucleotide diversity,while that of isolates from China,Turkey and Saudi Arabia was low.Strains from Turkey had the highest average difference number,indicating large genetic differences within the population compared to the other three countries where genetic differences were small.Haplotype diversity for isolates from Turkey,Iraq and Saudi Arabia was more than 0.5,with Iraqi isolates showing the highest value reflecting high genetic diversity,whereas Chinese strain value was 0.250 indicating low genetic diversity.Tajima’s D value for all four countries above were less than 0,suggesting that the population had shifted its genetic balance state,possibly due to population expansion.【Conclusion】 The data of this study showed that Theileria infection was prevalent in Keping and Awali counties,Aksu,and Theileria ovis was the main pathogen. Theileria uilenbergi and Theileria luwenshuni were not detected,and Theileria ovis had a distant genetic relationship with them.Theileria ovis populations from different regions were characterized by high haplotype and genetic diversity,and population expansion was occurring.The results of this study could provide a reference for the comprehensive control of ovine theileriosis and the healthy development of local aquaculture in Aksu.

Key words: sheep; Theileria; PCR detection; genetic evolution; prevention and control

中图分类号:

S852.72

周娜, 李才善, 赵雪晴, 阿布都卡迪尔·米吉提, 邓聿钤, 刘诗语, 石文玉, 郭庆勇, 巴音查汗·盖力克. 新疆阿克苏地区羊泰勒虫PCR检测及遗传多样性分析[J]. 中国畜牧兽医, 2025, 52(3): 1292-1300.

ZHOU Na, LI Caishan, ZHAO Xueqing, ABUDUKADIER· Mejiti, DENG Yuqian, LIU Shiyu, SHI Wenyu, GUO Qingyong, BAYINGCHAHAN· Gailike. PCR Detection and Genetic Diversity Analysis of Theileria of Sheep in Aksu,Xinjiang[J]. China Animal Husbandry and Veterinary Medicine, 2025, 52(3): 1292-1300.

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