Fredrick O. Obonyo, Ndichu Maingi, Samuel M. Githigia, Kevin O. Ochwedo, Anne A. Owiti, Evans N. Nyaboga
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Genotyping was performed on 15 selected positive samples using multilocus nested polymerase chain reaction restriction fragment length polymorphism (Mn-PCR-RFLP) with eight genetic markers (’SAG 2, 5’SAG 2, Alt. SAG 2, SAG 3, GRA 6, C29-2, BTUB and L358).</p><h3>Results</h3><p><i>Toxoplasma gondii</i> DNA was detected in 36.5% (35/96) of the blood samples. The sequences obtained exhibited 98.2–99.5% homology with those deposited in GenBank. Phylogenetic analysis demonstrated that the obtained sequences are conserved and clustered with those of infecting animals from other regions of the world. Eighteen distinct <i>T. gondii</i> haplotypes were identified to be circulating in donkeys from central Kenya. The <i>T. gondii</i> DNA samples exhibited high haplotype diversity (Hd: 0.915) and limited genetic diversity (π = 0.01027). PCR-RFLP of <i>T. gondii</i> DNA-positive samples revealed three different genetic combinations that consisted of alleles I, II and III, indicating the dissemination of atypical genotypes.</p><h3>Conclusion</h3><p>This study demonstrated that <i>T. gondii</i> is widespread in donkeys from Kenya and could be a possible source of infection in humans. These findings are important for designing control strategies for this parasite to improve the livestock sector, which is one of the main sources of livelihood for farmers in Kenya.</p></div>","PeriodicalId":6932,"journal":{"name":"Acta Parasitologica","volume":"69 3","pages":"1480 - 1491"},"PeriodicalIF":1.2000,"publicationDate":"2024-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s11686-024-00905-z.pdf","citationCount":"0","resultStr":"{\"title\":\"First Report on the Molecular Detection and Genetic Characterization of Toxoplasma gondii From Donkeys in Kenya\",\"authors\":\"Fredrick O. Obonyo, Ndichu Maingi, Samuel M. Githigia, Kevin O. Ochwedo, Anne A. Owiti, Evans N. 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引用次数: 0
摘要
目的：本研究旨在通过分子检测和对肯尼亚中部地区获得的 DNA 样本进行遗传多样性分析，确定驴体内是否存在弓形虫：方法：从梅鲁县和基里尼亚加县的驴子身上共采集了 363 份血样，其中 96 份血样曾用间接 ELISA 方法检测出弓形虫血清阳性，对其进行了基于内部转录间隔 1 (ITS-1) 基因扩增的巢式 PCR 检测，随后进行了 DNA 测序和系统发育分析。利用多焦点巢式聚合酶链反应限制性片段长度多态性（Mn-PCR-RFLP）和八个遗传标记（'SAG 2、5'SAG 2、Alt. SAG 2、SAG 3、GRA 6、C29-2、BTUB 和 L358）对 15 个选定的阳性样本进行了基因分型：结果：36.5%（35/96）的血液样本中检测到弓形虫 DNA。获得的序列与 GenBank 中的序列有 98.2-99.5% 的同源性。系统进化分析表明，所获得的序列与世界其他地区感染动物的序列是一致的，并且是聚类的。在肯尼亚中部地区的驴中发现了18种不同的淋病双球菌单倍型。淋球菌 DNA 样本表现出较高的单倍型多样性（Hd：0.915）和有限的遗传多样性（π = 0.01027）。淋球菌 DNA 阳性样本的 PCR-RFLP 显示出三种不同的基因组合，包括等位基因 I、II 和 III，表明非典型基因型的传播：这项研究表明，淋病双球菌广泛存在于肯尼亚的驴中，可能是人类的一个传染源。这些发现对于设计这种寄生虫的控制策略以改善作为肯尼亚农民主要生计来源之一的畜牧业非常重要。
First Report on the Molecular Detection and Genetic Characterization of Toxoplasma gondii From Donkeys in Kenya
Purpose
The present study was conducted to determine the presence of Toxoplasma gondii in donkeys by molecular tests and genetic diversity analysis of the obtained DNA samples from central Kenya.
Method
A total of 363 blood samples were collected from donkeys in Meru and Kirinyaga Counties, and 96 samples that were previously seropositive for T. gondii using indirect ELISA were subjected to nested PCR based on the amplification of the internal transcribed spacer 1 (ITS-1) gene followed by DNA sequencing and phylogenetic analysis. Genotyping was performed on 15 selected positive samples using multilocus nested polymerase chain reaction restriction fragment length polymorphism (Mn-PCR-RFLP) with eight genetic markers (’SAG 2, 5’SAG 2, Alt. SAG 2, SAG 3, GRA 6, C29-2, BTUB and L358).
Results
Toxoplasma gondii DNA was detected in 36.5% (35/96) of the blood samples. The sequences obtained exhibited 98.2–99.5% homology with those deposited in GenBank. Phylogenetic analysis demonstrated that the obtained sequences are conserved and clustered with those of infecting animals from other regions of the world. Eighteen distinct T. gondii haplotypes were identified to be circulating in donkeys from central Kenya. The T. gondii DNA samples exhibited high haplotype diversity (Hd: 0.915) and limited genetic diversity (π = 0.01027). PCR-RFLP of T. gondii DNA-positive samples revealed three different genetic combinations that consisted of alleles I, II and III, indicating the dissemination of atypical genotypes.
Conclusion
This study demonstrated that T. gondii is widespread in donkeys from Kenya and could be a possible source of infection in humans. These findings are important for designing control strategies for this parasite to improve the livestock sector, which is one of the main sources of livelihood for farmers in Kenya.
期刊介绍：
Acta Parasitologica is an international journal covering the latest advances in the subject.
Acta Parasitologica publishes original papers on all aspects of parasitology and host-parasite relationships, including the latest discoveries in biochemical and molecular biology of parasites, their physiology, morphology, taxonomy and ecology, as well as original research papers on immunology, pathology, and epidemiology of parasitic diseases in the context of medical, veterinary and biological sciences. The journal also publishes short research notes, invited review articles, book reviews.
The journal was founded in 1953 as "Acta Parasitologica Polonica" by the Polish Parasitological Society and since 1954 has been published by W. Stefanski Institute of Parasitology of the Polish Academy of Sciences in Warsaw. Since 1992 in has appeared as Acta Parasitologica in four issues per year.
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