Gonçalves Luiz Ricardo, Harrus Shimon, Gutiérrez Ricardo, Herrera Heitor Miraglia, de Souza Ramos Inalda Angélica, Porfírio Grasiela Edith de Oliveira, Nachum-Biala Yaarit, de Sousa Keyla Carstens Marques, da Silva Thiago Merighi Vieira, Campos João Bosco Vilela, Lemos Wagner, Moraes Barros-Battesti Darci, Machado Rosangela Zacarias, André Marcos Rogério
Graduate Program of Agricultural and Livestock Microbiology, Faculty of Agrarian and Veterinary Sciences (FCAV/UNESP), Jaboticabal, Brazil.
Department of Veterinary Pathology, Faculty of Agrarian and Veterinary Sciences (FCAV/UNESP), Jaboticabal, Brazil.
Transbound Emerg Dis. 2020 Sep;67(5):1888-1897. doi: 10.1111/tbed.13517. Epub 2020 Mar 10.
Currently, five Bartonella species and an expanding number of Candidatus Bartonella species have globally been reported in ruminants. Likewise, different Bartonella genotypes were identified. However, studies relating to ruminant-associated Bartonella in Brazil are scarce. The current study aimed to assess the prevalence and genetic diversity of Bartonella in cattle, buffaloes and associated ectoparasites in Brazil. For this purpose, EDTA-blood samples from 75 cattle and 101 buffaloes were sampled. Additionally, 128 Rhipicephalus microplus and one Amblyomma sculptum ticks collected from cattle, and 197 R. microplus, one A. sculptum and 170 lice (Haematopinus tuberculatus) collected from buffaloes were included. Bartonella DNA was initially screened through an HRM real-time PCR assay targeting the 16S-23S internal transcribed spacer (ITS), and the positive samples were submitted to an additional HRM assay targeting the ssrA gene. The HRM-positive amplicons were sequenced, and the nucleotide identity was assessed by BLASTn. Bartonella spp.-positive DNA samples were analysed by conventional PCR assays targeting the gltA and rpoB genes, and then, the samples were cloned. Finally, the phylogenetic positioning and the genetic diversity of clones were assessed. Overall, 21 of 75 (28%) cattle blood samples and 13 of 126 (10.3%) associated ticks were positive for Bartonella bovis. Out of 101 buffaloes, 95 lice and 188 tick DNA samples, one (1%) buffalo and four (4.2%) lice were positive for Bartonella spp. Conversely, none of the ticks obtained from buffaloes were positive for Bartonella. The Bartonella sequences from buffaloes showed identity ranging from 100% (ITS and gltA) to 94% (ssrA) with B. bovis. In contrast, the Bartonella DNA sequences from lice were identical (100%) to uncultured Bartonella sp. detected in cattle tail louse (Haematopinus quadripertusus) from Israel in all amplified genes. The present study demonstrates the prevalence of new B. bovis genotypes and a cattle lice-associated Bartonella species in large ruminants and their ectoparasites from Brazil. These findings shed light on the distribution and genetic diversity of ruminant- and ectoparasite-related Bartonella in Brazil.
目前，全球已在反刍动物中报告了5种巴尔通体属物种以及数量不断增加的暂定巴尔通体属物种。同样，也鉴定出了不同的巴尔通体基因型。然而，巴西关于反刍动物相关巴尔通体的研究很少。本研究旨在评估巴西牛、水牛及相关体外寄生虫中巴尔通体的流行情况和遗传多样性。为此，采集了75头牛和101头水牛的乙二胺四乙酸抗凝血样本。此外，还纳入了从牛身上采集的128只微小扇头蜱和1只刻点钝缘蜱，以及从水牛身上采集的197只微小扇头蜱、1只刻点钝缘蜱和170只虱子（结节血虱）。最初通过针对16S - 23S内部转录间隔区（ITS）的高分辨率熔解曲线实时聚合酶链反应（HRM real-time PCR）检测筛选巴尔通体DNA，阳性样本再进行针对ssrA基因的额外HRM检测。对HRM阳性扩增子进行测序，并通过BLASTn评估核苷酸同一性。通过针对gltA和rpoB基因的常规聚合酶链反应检测分析巴尔通体属阳性DNA样本，然后对样本进行克隆。最后，评估克隆的系统发育定位和遗传多样性。总体而言，75份牛血样本中有21份（28%）以及126只相关蜱中有13只（10.3%）对牛巴尔通体呈阳性。在101头水牛、95只虱子和188份蜱DNA样本中，有1头水牛（1%）和4只虱子（4.2%）对巴尔通体属呈阳性。相反，从水牛身上采集的蜱均未检测出巴尔通体阳性。水牛的巴尔通体序列与牛巴尔通体的同一性在100%（ITS和gltA）至94%（ssrA）之间。相比之下，虱子的巴尔通体DNA序列在所有扩增基因中与在以色列牛尾虱（四角血虱）中检测到的未培养巴尔通体种完全相同（100%）。本研究证明了巴西大型反刍动物及其体外寄生虫中存在新的牛巴尔通体基因型和一种与牛虱子相关的巴尔通体物种。这些发现揭示了巴西反刍动物和体外寄生虫相关巴尔通体的分布和遗传多样性。
