Campero Lucía María, Gos María Laura, Moore Dadin Prando, Regidor-Cerrillo Javier, Unzaga Juan Manuel, Moré Gastón, Ortega-Mora Luis Miguel, Venturini María Cecilia
Immunoparasitology Laboratory, School of Veterinary Sciences, National University of La Plata, 60 and 118 s/n, 1900 La Plata, Argentina; National Scientific and Technical Research Council, Godoy Cruz 2290, C1033AAJ Buenos Aires, Argentina.
National Scientific and Technical Research Council, Godoy Cruz 2290, C1033AAJ Buenos Aires, Argentina; Balcarce Agricultural Experimental Station, National Institute of Agricultural Technology, Ruta 226 Km 73.5, 7620 Balcarce, Argentina.
Vet Parasitol. 2018 May 15;255:58-60. doi: 10.1016/j.vetpar.2018.03.024. Epub 2018 Mar 28.
Neospora caninum is an apicomplexan protozoan responsible for abortion in ruminants. The present study aimed to diagnose an abortion from an Anglo Nubian goat from a dairy herd located in Buenos Aires province, Argentina. The goat delivered a fetus of approximately 3 months gestation that was studied by indirect fluorescent antibody test (IFAT), Immunoblot (IB), histopathology (HP), immunohistochemistry (IHC), and molecular assays (PCR, sequencing and microsatellite genotyping). Interferon gamma knock-out mice were inoculated with a pool of tissues for isolation attempts. The mother had IFAT titers of 1:3200 and 1:400 for N. caninum and Toxoplasma gondii, respectively, as well as positive IB reactions, whereas the fetus was seronegative to both parasites by IFAT and IB. The fetus had severe multifocal necrotizing myocarditis and hepatitis, moderate interstitial pneumonia, and nephritis. Myocardium sample resulted positive by IHC, evidencing clusters of N. caninum tachyzoites within myocardiocytes associated with histopathological lesions. Neospora caninum-DNA was detected by PCR in heart, liver, lungs, kidney, and muscle from the fetus, and was negative for T. gondii by PCR. NC-5 and 18 S rRNA gene fragment sequences showed 100% identity with N. caninum. Inoculated mice bled 30 days post-inoculation resulted seronegative to N. caninum and T. gondii by IFAT, and showed no clinical signs. Multilocus-microsatellite genotyping revealed a genetic profile that differed from previously reported N. caninum genotypes, with unique MS21 and MS10 alleles. These findings indicate that N. caninum was efficiently transmitted from the mother to the fetus. We report the first case of direct detection of N. caninum in a goat fetus in Argentina and N. caninum microsatellite genotyping in naturally infected goat.
犬新孢子虫是一种顶复门原生动物，可导致反刍动物流产。本研究旨在诊断一例来自阿根廷布宜诺斯艾利斯省一个奶牛场的安哥拉努比亚山羊的流产病例。这只山羊产下了一个妊娠约3个月的胎儿，对其进行了间接荧光抗体试验（IFAT）、免疫印迹（IB）、组织病理学（HP）、免疫组织化学（IHC）和分子检测（PCR、测序和微卫星基因分型）。为了尝试分离病原体，将一组组织接种到干扰素γ基因敲除小鼠体内。母羊的犬新孢子虫和刚地弓形虫的IFAT滴度分别为1:3200和1:400，且免疫印迹反应呈阳性，而胎儿通过IFAT和免疫印迹对这两种寄生虫均呈血清学阴性。胎儿患有严重的多灶性坏死性心肌炎和肝炎、中度间质性肺炎和肾炎。心肌样本经免疫组织化学检测呈阳性，显示心肌细胞内有犬新孢子虫速殖子簇，伴有组织病理学病变。通过PCR在胎儿的心脏、肝脏、肺、肾脏和肌肉中检测到犬新孢子虫DNA，而通过PCR检测弓形虫为阴性。NC-5和18 S rRNA基因片段序列与犬新孢子虫显示100%的同一性。接种后30天采血的小鼠通过IFAT对犬新孢子虫和弓形虫呈血清学阴性，且无临床症状。多位点微卫星基因分型显示出一种与先前报道的犬新孢子虫基因型不同的遗传图谱，具有独特的MS21和MS10等位基因。这些发现表明犬新孢子虫能够有效地从母体传播给胎儿。我们报告了阿根廷首例在山羊胎儿中直接检测到犬新孢子虫以及对自然感染山羊进行犬新孢子虫微卫星基因分型的病例。
