Genetic diversity and Multilocus Sequence Typing Analysis of Bartonella henselae in domestic cats from Southeastern Brazil
Genetic diversity and Multilocus Sequence Typing Analysis of Bartonella henselae in domestic cats from Southeastern Brazil
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DOI：
10.1016/j.actatropica.2021.106037
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发表时间：
2021-07-09
期刊：
影响因子：
2.7
通讯作者：
Andre, Marcos Rogerio
中科院分区：
文献类型：
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作者：
Chiaradia Furquim, Maria Eduarda;do Amaral, Renan;Andre, Marcos Rogerio
汉赛巴尔通体（Bartonella henselae）是传染病猫抓病（CSD）的病原体，其可以是致命的。已知家养和野生猫科动物是其主要的哺乳动物宿主。本研究的目的是调查巴尔通体的发生和遗传多样性。在巴西东南部的圣保罗（SP）和米纳斯吉拉斯（MG）州采集的猫样本中。基于定量实时PCR（qPCR）分析，在39.9%（122/306）的血液样本（SP的46/151只猫; MG的76/155只猫）中检测到巴尔通体nuoG基因片段。将血液样本提交至预富集培养技术，该技术允许检测12个额外的阳性样本，这些样本在使用DNA血液样本作为模板的qPCR中显示为阴性。此外，5个B。从用于血液和预富集培养的qPCR阴性样品中获得汉赛氏菌分离株。24只猫栉首蚤中有7只巴尔通体阳性。在qPCR试验中;从巴尔通体阴性猫中收集了4/7只阳性跳蚤。23转B B。从九只猫的血液样本中获得了henselae克隆序列，显示存在13种不同的基因型。Median-joining network和SplitsTree距离分析表明，获得的序列代表了明显的B。henselae基因型进行比较。由于B的rpo B基因型不同，在寄主内存在多样性。在单个猫中检测到汉赛氏菌。对汉赛巴尔通体8个分子标记进行多位点序列分型，结果显示两个等位基因型（MG状态下的ST 37和SP状态下的ST 9）。本研究是第一次从米纳斯吉拉斯州的猫中发现巴尔通体的分子报告。总之，这部分工作显示了不同B的发生。henselae rpoB基因型在水库内宿主水平。基于血液样本和预增菌液体培养和分离的qPCR，在SP和MG状态的猫中分别发现33.1%（50/151）和56.8%（88/155）的巴尔通体属。B的两种不同等位基因谱。在圣保罗州（ST 9）和米纳斯吉拉斯州（ST 37）的猫中发现了henselae，表明Bartonelae在一定地理区域内发生了克隆进化。
Bartonella henselae is the causative agent for the infectious disease Cat Scratch Disease (CSD), which can be fatal. Domestic and wild felines are known to be its main mammal reservoirs. The present study aimed to investigate the occurrence and genetic diversity of Bartonella spp. in cats sampled in Sao Paulo (SP) and Minas Gerais (MG) States, Southeastern Brazil. Based on a quantitative real-time PCR (qPCR) assay, a Bartonella sp. nuoG gene fragment was detected in 39.9% (122/306) of the blood samples (46/151 cats of SP; 76/155 cats of MG). The blood samples were submitted to a pre-enrichment culture technique that allowed the detection of 12 additional positive samples, which showed to be negative in the qPCR using DNA blood samples as templates. Furthermore, five B. henselae isolates were obtained from qPCR-negative samples for both blood and pre-enrichment culture. Seven out of 24 Ctenocephalides felis fleas were positive for Bartonella spp. in the qPCR assay; 4/7 positive fleas were collected from Bartonella-negative cats. Twenty-three rpoB B. henselae cloned sequences were obtained from nine cats' blood samples, showing the occurrence of 13 different genotypes. Median-joining network and SplitsTree distance analysis showed that the obtained sequences represented distinct B. henselae genotypes when compared to those previously deposited in GenBank. Intra-host diversity was found, since different rpoB genotypes of B. henselae were detected in individual single cats. Bartonella henselae isolates showed two allelic profiles (ST37 in cats from MG state and ST9 in SP state) by MLST (Multilocus Sequence Typing) based on sequencing of eight molecular markers. The present study is the first molecular report of Bartonella sp. in cats from Minas Gerais State. In summary, this body of work showed the occurrence of different B. henselae rpoB genotypes at an intra-reservoir host level. Based on qPCR from blood samples and pre-enrichment liquid culture and isolation, occurrence of 33.1% (50/151) and 56.8% (88/155) for Bartonella sp. was found in cats from SP and MG states, respectively. Two different allelic profiles of B. henselae were found in cats from the states of Sao Paulo (ST9) and Minas Gerais (ST37), suggesting a clonal evolution of Bartonellae in a certain geographical region.