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Prevalence and genotyping of Toxoplasma gondii in stray cats in Mashhad area, Iran

机译:伊朗Mashhad地区流浪猫弓形猫的患病率和基因分型

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BACKGROUND:Cats as a definitive host have an important role in the epidemiology of toxoplasmosis in humans and animals. The aim of the study was to determine the frequency of Toxoplasma gondii infection and isolate and identify the genotypes of T. gondii in stray cats in the Mashhad suburb.METHODS:From April 2016 to August 2017, 175 fecal samples from stray cats and 31 brain samples from cats killed in driving accidents were collected. The fecal samples were examined by fecal flotation technique and T. gondii-specific PCR. The brain samples were investigated by T. gondii-specific PCR and consequently examined by mice bioassay. The DNA of T. gondii isolated was genotyped using SAG1, SAG2, SAG3, BTUB and GRA6 as PCR-restriction fragment length polymorphism (PCR-RFLP) markers.RESULTS:In the present study, Toxoplasma-like oocysts were microscopically observed in 2.2% (4/175) fecal samples. The presence of Toxoplasma oocysts was confirmed in one microscopy-positive sample by PCR. In addition, T. gondii DNA was detected in 4% (7/175) microscopy-negative samples using PCR. T. gondii was isolated from one brain PCR-positive sample by mice bioassay. The isolate was avirulent and many T. gondii cysts were observed in mice brain. The isolate was successfully genotyped by PCR-RLFP analysis. The isolated genotyped was type II. Besides, eight Toxoplasma-positive fecal samples contained insufficient DNA and only amplified at SAG-3 locus in PCR. These samples were also showed type II pattern at this locus.CONCLUSIONS:Parasitological and molecular results showed low frequency of Toxoplasma infection in the stray cats, and identified the genotype of T. gondii isolate as type II, for the first time in Mashhad area, Khorasan Razavi Province.
机译:背景:作为最终宿主的猫对人类和动物的弓形虫病的流行病学具有重要作用。该研究的目的是确定弓形虫感染的频率和孤立,并鉴定MASHHAD郊区的流浪猫中T.Gondii的基因型。方法:从2016年4月到2017年8月,来自杂志和31例脑的175个粪便样本收集来自驾驶事故中的猫的样品。通过粪便浮选技术和T.Gondii特异性PCR检查粪便样品。通过T.Gondii特异性PCR研究了脑样品,并因此通过小鼠生物测定来研究。分离的T.Gondii的DNA使用SAG1,SAG2,SAG3,BTUB和GRA6基因分型作为PCR限制性片段长度多态性(PCR-RFLP)标记。结果:在本研究中,在2.2%的情况下显微镜观察到弓形虫卵囊。 (4/175)粪便样本。通过PCR在一个显微镜阳性样品中证实弓形虫卵囊的存在。此外,使用PCR在4%(7/175)显微镜阴性样品中检测到Gondii DNA。通过小鼠生物测定法从一个脑PCR阳性样品中分离出来的Gondii。分离物是无毒,在小鼠脑中观察到许多巨杆菌囊肿。通过PCR-RLFP分析成功基因分离分离。分离的基因分型是II型。此外,八个弓形阳性粪便样品含有不足的DNA,并且仅在PCR中的SAG-3位点扩增。这些样品也在该基因座中显示II型图案。结论:寄生学和分子结果显示杂散猫类中的弓形虫感染频率低,并鉴定了T.Gondii孤立的基因型,如II型,第一次在MASHHAD地区首次, Khorasan Razavi省。

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