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犬血清对犬神经肉孢子虫和克鲁斯肉孢子虫抗原的血清学反应性。

Serologic reactivity of canine sera to Sarcocystis neurona and Sarcocystis cruzi antigens.

作者信息

Oliveira Cleusa Maria Carneiro, Oliveira Philipe Brito, Albuquerque George Rego, Gondim Luís Fernando Pita

机构信息

Departamento de Anatomia, Patologia e Clínicas, Universidade Federal da Bahia, Escola de Medicina Veterinária e Zootecnia, Avenida Adhemar de Barros, 500, Ondina, Salvador 40170-110, Bahia, Brazil.

Departamento de Ciências Agrárias e Ambientais, Universidade Estadual de Santa Cruz, Ilhéus, Bahia, Brazil.

出版信息

Vet Parasitol Reg Stud Reports. 2020 Jul;21:100439. doi: 10.1016/j.vprsr.2020.100439. Epub 2020 Jul 22.

Abstract

Sarcocystis neurona, a coccidian parasite shed by opossums (Didelphis spp.) in the Americas, is the major cause of equine protozoal myeloencephalitis (EPM) and induces disease in other domestic and wild animal species, including domestic dogs. Sarcocystis cruzi, despite its low pathogenicity for cattle (intermediate hosts), is worldwide distributed and uses mostly dogs as definitive hosts. The aims of this study were to test serological reactivities of dog sera to S. neurona and S. cruzi antigens, and to investigate potential serological cross-reactivity to these parasites. Sera from 353 Brazilian dogs were obtained from rural areas in the municipality of Ilhéus, Bahia, and examined by immunofluorescent antibody tests (IFAT). Antigens used in serological reactions consisted of S. neurona merozoites from a North American strain (SN138), and bradyzoites of S. cruzi obtained from Brazilian bovine hearts, with parasite species identity confirmed by PCR and sequencing of the 18S gene of the rDNA. Seropositivity to S. neurona and to S. cruzi were detected in 3.39% (12/353) and 4.81% (17/353) of the dogs, respectively. Ten canine sera reacted solely to S. neurona and 15 serum samples reacted only to S. cruzi. Two serum samples were simultaneously positive for both parasites. Sera from 14 dogs that tested positive by IFAT (9 for S. neurona and 3 for S. cruzi) and from two dogs that were negative by IFAT for the two parasites, were examined by Western blot using S. neurona as antigen; these sera reacted to a great number of protein bands, including antigens on the 16 and 30 KDa positions, which encompass immunodominant antigens for S. neurona in horses. Western blot did not show any specific pattern for S. neurona infection/exposure using canine sera. Dogs act as definitive hosts for several Sarcocystis spp. that infect farm animals, including horses, sheep, goats, water buffaloes and pigs, and for this reason, should contain antibodies to a broad repertoire of Sarcocystis spp. antigens. In conclusion, low percentages of dogs from rural areas of Ilhéus, Bahia, were reactive to both S. neurona and S. cruzi antigens. It is possible that other Sarcocystis species, besides S. neurona and S. cruzi, might have contributed for the seropositivity observed in this study. IFAT was more specific than Western blot to differentiate canine serological reactions to S. neurona and S. cruzi antigens.

摘要

肉孢子虫属神经元种是一种球虫寄生虫,由美洲负鼠(绵毛负鼠属)排出,是马属动物原虫性脑脊髓炎(EPM)的主要病因,并可在包括家犬在内的其他家养和野生动物物种中引发疾病。克氏肉孢子虫尽管对牛(中间宿主)致病性较低,但分布于全球,主要以犬作为终末宿主。本研究的目的是检测犬血清对肉孢子虫属神经元种和克氏肉孢子虫抗原的血清学反应性,并调查对这些寄生虫的潜在血清学交叉反应性。从巴伊亚州伊列乌斯市农村地区的353只巴西犬采集血清,通过免疫荧光抗体试验(IFAT)进行检测。血清学反应中使用的抗原包括来自北美毒株(SN138)的肉孢子虫属神经元种裂殖子,以及从巴西牛心脏获得的克氏肉孢子虫缓殖子,通过PCR和rDNA的18S基因测序确认寄生虫种类。分别在3.39%(12/353)和4.81%(17/353)的犬中检测到对肉孢子虫属神经元种和克氏肉孢子虫的血清阳性。10份犬血清仅对肉孢子虫属神经元种有反应,15份血清样本仅对克氏肉孢子虫有反应。两份血清样本对两种寄生虫均呈阳性。对IFAT检测呈阳性的14只犬(9只对肉孢子虫属神经元种阳性,3只对克氏肉孢子虫阳性)以及对两种寄生虫IFAT检测均为阴性的两只犬的血清,以肉孢子虫属神经元种为抗原进行蛋白质印迹分析;这些血清与大量蛋白条带发生反应,包括16和30 kDa位置的抗原,这些抗原包含马属动物中肉孢子虫属神经元种的免疫显性抗原。蛋白质印迹分析未显示使用犬血清检测肉孢子虫属神经元种感染/暴露的任何特定模式。犬作为几种感染农场动物(包括马、绵羊、山羊、水牛和猪)的肉孢子虫属物种的终末宿主,因此,应含有针对多种肉孢子虫属物种抗原的抗体。总之,巴伊亚州伊列乌斯市农村地区的犬中,对肉孢子虫属神经元种和克氏肉孢子虫抗原呈反应性的比例较低。除肉孢子虫属神经元种和克氏肉孢子虫外,其他肉孢子虫属物种可能导致了本研究中观察到的血清阳性。在区分犬对肉孢子虫属神经元种和克氏肉孢子虫抗原的血清学反应方面,IFAT比蛋白质印迹分析更具特异性。

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