Ticks and tick-borne pathogens are one of the major threats to livestock production worldwide. The aim of present study is to specify the molecular epidemiology and its associated risk factors of two well-distributed tick-borne pathogens Anaplasma marginale and Theileria annulata in cattle population from North-western Pakistan. Blood samples were Collected from 60 (32.6 %) local breed cattle, 101 (54.9 %) from crossbreed, and 24 (13.0 %) from exotic breed with total of 184 blood samples. Species-specific PCR assays were performed to detect the presence of A. marginale and T. annulata based on 16S rRNA and 18S rRNA genetic makers respectively. PCR results showed that A. marginale was 16.3 % prevalent and T. annulata was 29.9 % prevalent in the study area with a total prevalence rate of 46.2 % (85/184) of the tested blood samples. District wise analysis revealed that both pathogens were highly prevalent in district Dir Upper (13.6 %) and least prevalent in district Chitral (10.3 %). Univariable analysis of risk factors showed that only breed and acaricidal treatment were significant determinants (P < 0.05) for A. marginale infection, however, in case of T. annulata infection; breed, age, gender, grazing practice, and acaricidal treatment were potential determinants (P < 0.05). Multivariable analysis specified that breed and acaricidal treatment were considered as significant risk factors for A. marginale infection (P < 0.05) whereas acaricidal treatment was found to be a significant determinant for T. annulata infection (P < 0.05). Phylogenetic analysis indicated that A. marginale 16S rRNA and T. annulata 18S rRNA isolates showed similarities and shared phylogeny with same isolates reported from Asia. This is the first molecular report on the epidemiology and risk factors analysis of A. marginale and T. annulata infections in cattle population from NW Pakistan. Further large scale study is required to investigate molecular, epidemiological and genotypic aspects as well as potential risk factors analysis from the country to facilitate designing strategies to control tick-borne pathogen and reduce losses to cattle industry.

中文翻译：

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巴基斯坦西北部牛的无性无浆体和无环泰勒菌的分子流行病学及相关危险因素。

壁虱和壁虱传播的病原体是全球牲畜生产的主要威胁之一。本研究的目的是详细说明巴基斯坦西北部牛群中两种分布均匀的tick传病原体Anaplasma marginale和Theileria annulata的分子流行病学及其相关的危险因素。从60头（32.6％）本地牛，101头（54.9％）杂种和24头（13.0％）的外来品种中采集血液样本，总共184个血液样本。分别基于16S rRNA和18S rRNA遗传标记物进行物种特异性PCR检测，以检测边缘缘农杆菌和环线虫的存在。PCR结果显示，在研究区域中，拟南芥的患病率为16.3％，而环纹菌的患病率为29.9％，被测血样的总患病率为46.2％（85/184）。地区明智的分析显示，两种病原体在上迪尔上游地区（13.6％）高度流行，而在Chitral地区（10.3％）最不流行。危险因素的单因素分析表明，仅品种和杀螨剂处理是边缘margin曲霉感染的重要决定因素（P <0.05），而在无环线虫感染的情况下；品种，年龄，性别，放牧习惯和杀螨剂处理是潜在的决定因素（P <0.05）。多变量分析表明，品种和杀螨剂处理被认为是A.marginale感染的重要危险因素（P <0.05），而杀螨剂处理被认为是环孢菌感染的重要决定因素（P <0.05）。系统发育分析表明，A.边缘16S rRNA和T. 环孢菌18S rRNA分离株显示出相似性，并且与亚洲报道的相同分离株具有相同的系统发育。这是关于巴基斯坦西北部牛群中margin缘和环线虫感染的流行病学和危险因素分析的第一份分子报告。该国需要进一步的大规模研究，以调查分子，流行病学和基因型方面，以及来自该国的潜在危险因素分析，以促进设计策略来控制pathogen传播的病原体并减少对牛业的损失。