Isolation and Genotyping of Toxoplasma gondii Strains in Ovine Aborted Fetuses in Khorasan Razavi Province, Iran

Toxoplasmosis is an important zoonotic disease that can cause abortion in humans and animals. The aim of this study was isolation and subsequent genotyping of Toxoplasma gondii isolates in ovine aborted fetuses. During 2012-2013, 39 ovine aborted fetuses were collected from sheep flocks in Khorasan Razavi Province, Iran. The brain samples were screened for detection of the parasite DNA by nested PCR. The positive brain samples were bioassayed in Webster Swiss mice. The serum samples of mice were examined for T. gondii antibodies by IFAT at 6 weeks post inoculation, and T. gondii cysts were searched in brain tissue samples of seropositive mice. The positive samples were genotyped by using a PCR-RLFP method. Subsequently, GRA6 sequences of isolates were analyzed using a phylogenetic method. The results revealed that T. gondii DNA was detected in 54% (20/37, 95% CI 38.4-69.0%) brain samples of ovine aborted fetuses. In bioassay of mice, only 2 samples were virulent and the mice were killed at 30 days post inoculation, while the others were non-virulent to mice. The size of cysts ranged 7-22 µm. Complete genotyping data for GRA6 locus were observed in 5 of the 20 samples. PCR-RLFP results and phylogenetic analysis revealed that all of the isolated samples were closely related to type I. For the first time, we could genotype and report T. gondii isolates from ovine aborted fetuses in Khorasan Razavi Province, Iran. The results indicate that the T. gondii isolates are genetically related to type I, although most of them were non-virulent for mice.

[Molecular identification of leishmania species causing cutaneous leishmaniasis in Mashhad area, Iran]

Background and Objective: Cutaneous leishmaniasis is an endemic diseases with important public health in Iran. The cinical signs of disease were seen as dry and wet forms. It is essential to distinguish leishmania species in every area for designing control of diseases. The morphology of leishman bodies in each speiecis are very similar and need to sensitive diagnostic method such as PCR to differntiate of them. The present study, a molecular study has been done to identify of leishmania species in Mashhad area from autumn in 2011 to summer in 2013

Methods: Firstly, tissue smears were collected from the lesion of one hundred patients and from the ulcer of 25 paitents were cultured in NNN nutrified with RPMI. The positive samples [tissue smears and culture] were used for DNA extraction and PCR. PCR methods were used in two steps. Firstly a sensitive PCR was used to detect the leishmania genus and secondly, the positive samples were examined with species specific semi-nested - PCR

Results: In first step of PCR, all of sampled were positive for Leishmania spp and in second step Leishmania tropica and L.major were detected in 94% and 6% in positive -PCR amplicon, respectively

Conclusion: Based on the results, Leishmania tropica is more prevalent than L.major in Mashhad area