Molecular Detection of Anaplasma phagocytophilum in Ixodid Ticks Infesting White Fulani Cattle in Zaria and its Environs, Kaduna State, Nigeria.

M. Rabiu, A. Natala, O. O. Okubanjo, R. David, S. Ola-Fadunsin, K. Hussain, I. Ganiyu, Ismail Ayoade Odetokun, N. Elelu, D. Sani, A. Y. Baba
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Abstract

Anaplasmosis, a disease caused by various species of Anaplasma, poses important economic constraints to animal breeders. In Zaria, molecular detection of tick-borne pathogens infecting cattle is very sketchy. Being hematophagous, ticks are capable of transmitting disease agents such as viruses, bacteria and protozoa. This study was carried out to detect for the presence of Anaplasma pathogen in the various tick species infesting white Fulani breeds of cattle owing to their high population in Zaria and environs using a Polymerase Chain Reaction (PCR). Three hundred and eighty-four (n=384) white Fulani breed of cattle of varying age and sex were sampled from 32 herds in 4 villages (Bomo, Tofu, Ungwan Dabosa and Majeru). The DNA was extracted using Qiagen commercial kit following manufacturer’s instructions. Genomic DNA of the species of ticks was amplified in a semi-nested PCR targeting the 16S rRNA fragment of the Anaplasma spp at expected amplicon size of 711bp. The PCR products were purified using Gel Extraction Kit (Bioland, Scientific LLC) following the manufacturer’s protocol. All sequences were subjected to a Basic Local Alignment Search Tool (BLAST) to determined their identities and assess their homologues and similarities to those in the GenBank. A neighbor-joining tree was generated using the Molecular Engineering Genetic Analysis (MEGA 7.0) to derive the close relationship between the isolates. The result of this study implies that Anaplasma spp (MN044909) is present in all the five species of ticks infesting cattle in Zaria, Kaduna State, Nigeria and pose a high risk on humans and animals health. Anaplasmosis, a disease caused by various species of Anaplasma, poses important economic constraints to animal breeders. In Zaria, molecular detection of tick-borne pathogens infecting cattle is very sketchy. Being hematophagous, ticks are capable of transmitting disease agents such as viruses, bacteria and protozoa. This study was carried out to detect for the presence of Anaplasma pathogen in the various tick species infesting white Fulani breeds of cattle owing to their high population in Zaria and environs using a Polymerase Chain Reaction (PCR). Three hundred and eighty-four (n=384) white Fulani breed of cattle of varying age and sex were sampled from 32 herds in 4 villages (Bomo, Tofu, Ungwan Dabosa and Majeru). The DNA was extracted using Qiagen commercial kit following manufacturer’s instructions. Genomic DNA of the species of ticks was amplified in a semi-nested PCR targeting the 16S rRNA fragment of the Anaplasma spp at expected amplicon size of 711bp. The PCR products were purified using Gel Extraction Kit (Bioland, Scientific LLC) following the manufacturer’s protocol. All sequences were subjected to a Basic Local Alignment Search Tool (BLAST) to determined their identities and assess their homologues and similarities to those in the GenBank. A neighbor-joining tree was generated using the Molecular Engineering Genetic Analysis (MEGA 7.0) to derive the close relationship between the isolates. The result of this study implies that Anaplasma spp (MN044909) is present in all the five species of ticks infesting cattle in Zaria, Kaduna State, Nigeria and pose a high risk on humans and animals health.
尼日利亚卡杜纳州扎里亚及其周边地区白富拉尼牛感染伊蚊蜱无原体嗜吞噬细胞分子检测
无形体病是一种由多种无形体引起的疾病,对动物饲养者造成了重要的经济限制。在扎里亚,对感染牛的蜱传病原体的分子检测非常粗略。作为食血动物,蜱虫能够传播病毒、细菌和原生动物等疾病媒介。本研究采用聚合酶链式反应(PCR)技术,对扎里亚及其周边地区富拉尼白牛种群众多的各种蜱类进行无原体病原体检测。从4个村(Bomo、Tofu、Ungwan Dabosa和Majeru)的32个畜群中采集了384头不同年龄和性别的白色富拉尼牛。按照制造商的说明,使用Qiagen商用试剂盒提取DNA。采用半巢式PCR方法,以无原体的16S rRNA片段为目标,扩增出蜱的基因组DNA,扩增子大小为711bp。PCR产物使用凝胶提取试剂盒(Bioland, Scientific LLC)按照制造商的协议纯化。所有序列都经过基本局部比对搜索工具(BLAST)来确定它们的身份,并评估它们与GenBank中的同源物和相似性。利用分子工程遗传分析软件MEGA 7.0建立了相邻连接树,分析了菌株间的亲缘关系。这项研究的结果表明,在尼日利亚卡杜纳州扎里亚(Zaria)感染牛的所有五种蜱虫中都存在无原体spp (MN044909),并对人类和动物健康构成高风险。无形体病是一种由多种无形体引起的疾病,对动物饲养者造成了重要的经济限制。在扎里亚,对感染牛的蜱传病原体的分子检测非常粗略。作为食血动物,蜱虫能够传播病毒、细菌和原生动物等疾病媒介。本研究采用聚合酶链式反应(PCR)技术,对扎里亚及其周边地区富拉尼白牛种群众多的各种蜱类进行无原体病原体检测。从4个村(Bomo、Tofu、Ungwan Dabosa和Majeru)的32个畜群中采集了384头不同年龄和性别的白色富拉尼牛。按照制造商的说明,使用Qiagen商用试剂盒提取DNA。采用半巢式PCR方法,以无原体的16S rRNA片段为目标,扩增出蜱的基因组DNA,扩增子大小为711bp。PCR产物使用凝胶提取试剂盒(Bioland, Scientific LLC)按照制造商的协议纯化。所有序列都经过基本局部比对搜索工具(BLAST)来确定它们的身份,并评估它们与GenBank中的同源物和相似性。利用分子工程遗传分析软件MEGA 7.0建立了相邻连接树,分析了菌株间的亲缘关系。这项研究的结果表明,在尼日利亚卡杜纳州扎里亚(Zaria)感染牛的所有五种蜱虫中都存在无原体spp (MN044909),并对人类和动物健康构成高风险。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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