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.

Detection and identification of Toxoplasma gondii type one infection in sheep aborted fetuses in Qazvin province of Iran

The aim of this study was to apply the nested-PCR and bioassay methods in detection and genotyping of Toxoplasma gondii infection in provided sheep aborted fetus samples from Qazvin Province of Iran. Eighteen sheep aborted fetal samples were studied by nested-PCR-RFLP, histopathological observation and microbiological assay. Bioassay in mice was carried out by inoculating the brain samples intraperitoneally. The results demonstrated the frequency of 66% infected sheep aborted fetal samples with T. gondii type one. Although we could not isolate any parasite from inoculated mice even after three passages, but it was confirmed histopathologically formation of cyst like bodies in prepared mice brain sections. The results of the performed nested-PCR and formation of brain cyst in inoculated mice exhibited that T. gondii type one infection might be considered as one of the major causative agents for abortion in ewes

Molecular and pathological study of bovine aborted fetuses and placenta from Neospora caninum infected dairy cattle

The objective of the study was to evaluate the presence of Neospora caninum organisms in the brain of aborted fetuses and placentas of full-term calves born of seropositive cows. During 2006-2007, 12 brains of aborted calves from Neospora seropositive cattle and 7 placentas from seropositive dams giving birth to full-term calves, from four dairy cattle farms located around Tehran province, Iran were examined by Nested-PCR and histopathology techniques. The Nested-PCR demonstrated that all of 12 aborted fetal brain samples and 5 of 7 placentas were infected by N. caninum. Mild to severe placentitis was observed in 5 placentas. Severe hyperemia and perivascular and perineuronal edema revealed in all fetal brain. In 3 out of 12 brains, scattered foci of hemorrhages, neuropilar necrosis and gliosis were present. In addition, nonpurulent encephalitis with severe lymphohistiocytic perivascular cuffing in one case and a small tissue cyst like Neospora caninum cyst in other calf were observed. Our results confirmed the molecular and histopathologic findings of other studies about Neospora caninum infection and it seems to support the hypothesis that Neospora infection is associated with bovine abortion in Iran