Feitosa Thais Ferreira, Vilela Vinícius Longo Ribeiro, de Almeida-Neto João Leite, de Melo Lídio Ricardo Bezerra, de Morais Dayana Firmino, Alves Bruna Farias, Nakashima Fabiana, Gennari Solange Maria, Athayde Ana Célia Rodrigues, de Jesus Pena Hilda Fátima
Federal University of Campina Grande (UFCG), Patos, PB, CEP 58708-110, Brazil.
Department of Veterinary Medicine, Federal Institute of Paraíba (IFPB), Sousa, PB, CEP 58800-970, Brazil.
Parasitol Res. 2017 Aug;116(8):2265-2270. doi: 10.1007/s00436-017-5531-x. Epub 2017 Jun 21.
This study evaluated, for the first time, the genetic diversity of Toxoplasma gondii isolates from free-range chickens from the state of Paraíba, Northeast Brazil. Tissue samples from 33 chickens from properties in five municipalities of Paraíba (Esperança, Olho d'Água, Malta, Monteiro, and Patos) were bioassayed in mice. The brains of mice infected with T. gondii cysts were used for DNA extraction and genotyping. Genotyping was performed using 11 PCR-RFLP markers and 15 microsatellite (MS) markers. Complete genotyping results were obtained for 29 isolates, with nine genotypes detected by RFLP and 15 genotypes identified by MS. Three genotypes (#273, #274, and #277) have only been recently identified from pigs in the region. Brazilian clonal types BrII and BrIII were identified from one isolate each. Clonal types I, II, and III were not detected by RFLP. Genotype #13 (Caribbean 1), detected in 48.3% (14/29) of isolates from four of the five municipalities investigated, was the most prevalent genotype in the state of Paraíba. However, the MS analysis showed that of these 14 isolates, only four were unique genotypes, and considering the distance between the municipalities from where they were collected, it is possible that only seven are independent isolates while the others are clones. The other genotypes were restricted to different microregions. The results indicate that the Caribbean 1 lineage of T. gondii is circulating widely in Northeast Brazil. The genotypic diversity of T. gondii in the state of Paraíba is high, and microsatellite analysis revealed this diversity with higher resolution than PCR-RFLP.
本研究首次评估了来自巴西东北部帕拉伊巴州散养鸡的刚地弓形虫分离株的遗传多样性。从帕拉伊巴州五个市（埃斯佩兰萨、奥约德阿瓜、马尔塔、蒙泰罗和帕托斯）的养殖场采集了33只鸡的组织样本，并在小鼠中进行了生物测定。用感染了刚地弓形虫包囊的小鼠大脑进行DNA提取和基因分型。使用11个PCR-RFLP标记和15个微卫星（MS）标记进行基因分型。对29个分离株获得了完整的基因分型结果，通过RFLP检测到9种基因型，通过MS鉴定出15种基因型。三种基因型（#273、#274和#277）最近才在该地区的猪中被鉴定出来。分别从一个分离株中鉴定出巴西克隆型BrII和BrIII。RFLP未检测到克隆型I、II和III。在调查的五个市中的四个市的48.3%（14/29）的分离株中检测到的基因型#13（加勒比1型）是帕拉伊巴州最常见的基因型。然而，MS分析表明，在这14个分离株中，只有4个是独特的基因型，考虑到采集它们的市之间的距离，有可能只有7个是独立的分离株，而其他的是克隆株。其他基因型局限于不同的微区域。结果表明，刚地弓形虫的加勒比1谱系在巴西东北部广泛传播。帕拉伊巴州刚地弓形虫的基因型多样性很高，微卫星分析比PCR-RFLP更能高分辨率地揭示这种多样性。
