Isolation and genetic characterization of Toxoplasma gondii from chickens from public markets in Pernambuco State, Brazil

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Abstract

This study aimed to evaluate the presence and viability of Toxoplasma gondii in chickens intended for human consumption in the Pernambuco State, Brazil. Blood and tissue samples were collected from 25 chickens sold in markets in Recife, Pernambuco. Samples were evaluated by indirect immunofluorescence assay (IFA) to detect antibodies to T. gondii. Pools of brain and heart of seropositive chickens were subjected to bioassay in two Swiss Webster mice, which were evaluated for 45 days then tested by IFA to detect seroconversion. The mice were euthanized, and their brains were evaluated for cysts. Peritoneal lavage was also conducted in mice that exhibited clinical signs. Brains containing cysts or peritoneal lavage with tachyzoites were inoculated into MA-104 cells. Brains of mice inoculated with the same tissue were pooled and analysed by ITS1-PCR. We obtained a frequency of antibodies to T. gondii of 68.00% (17/25) in chickens, and a seroconversion rate of 70.58% (24/34) in mice. Detection of Toxoplasma ITS1 DNA confirmed an isolation rate of 41.1% (7/17). Three isolates were characterized by mnPCR-RFLP as genotypes ToxoDB#36 and ToxoDB#114. We highlight the occurrence of ToxoDB#36 in chickens in Pernambuco State and the parasitesʼ viability in chickens intended for human consumption.

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从巴西伯南布哥州公共市场的鸡中分离出刚地弓形虫并确定其遗传特征
本研究旨在评估巴西伯南布哥州供人类食用的鸡体内弓形虫的存在和存活情况。研究人员从伯南布哥州累西腓市场上出售的 25 只鸡身上采集了血液和组织样本。样本通过间接免疫荧光试验(IFA)进行评估,以检测淋病双球菌抗体。血清反应阳性鸡的大脑和心脏样本在两只瑞士韦伯斯特小鼠体内进行生物测定,对小鼠进行 45 天的评估,然后用间接免疫荧光法检测血清转换。小鼠安乐死后,对其大脑进行囊肿评估。还对出现临床症状的小鼠进行腹腔灌洗。将含有囊肿或腹腔灌洗液中含有速殖体的大脑接种到 MA-104 细胞中。将接种了相同组织的小鼠大脑集中起来,通过 ITS1-PCR 进行分析。我们在鸡体内获得了68.00%(17/25)的弓形虫抗体,在小鼠体内获得了70.58%(24/34)的血清转换率。弓形虫 ITS1 DNA 的检测证实分离率为 41.1%(7/17)。三个分离株通过 mnPCR-RFLP 鉴定为基因型 ToxoDB#36 和 ToxoDB#114。我们强调了 ToxoDB#36 在伯南布哥州鸡只中的出现以及寄生虫在供人类食用的鸡只中的存活率。
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