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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.

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