Investigation of Toxoplasma gondii, Neospora caninum and Tritrichomonas foetus in abortions of cattle, sheep and goats in Turkey: Analysis by real-time PCR, conventional PCR and histopathological methods.

The aim of this study was to identify Neosopora caninum, Toxoplasma gondii and Tritricomonas foetus in all cattle aborted fetus samples and N. caninum and T. gondii in sheep and goat aborted fetuses sent to Elazig Veterinary Control Institute during two years. Total genomic DNAs were obtained using a commercial kit. Real-time PCR analysis was performed separately for each agent. Conventional PCR was set up for confirmation of positive samples. Then, fetal brain, heart, lung and liver samples were analysed by hematoxylin-eosin (HE) and Avidin-Biotin Complex (ABC) Immunohistochemistry (IHC) methods. Totally, we tested 55 aborted fetus samples. Of these samples, seven (12.7 %) was belonged to goats, 18 (32.7 %) to sheep and 30 (54.5 %) to cattle. T. gondii was detected in six (10.90 %) samples, and four (7.27 %) of them were positive with Real-time PCR, while only one of these four samples was positive for both classical PCR and IHC. N. caninum was determined by at least one of the three tests in 14 (25.45 %) of the samples studied, while 8 (14.54 %) of the positive samples were detected by Real-time PCR, only two of them were also positive with conventional PCR, eight (14.54 %) samples was determined as positive by IHC. Considering T. foetus in the samples, positivity was determined in two (3.63 %) of 55 aborted fetus (both of which were aborted cattle fetus) by Real-time PCR, while only one of them was positive with conventional PCR, while no positivity was detected with the IHC.

Serological and Molecular Detection of Dirofilaria Species in Stray Dogs and Investigation of Wolbachia DNA by PCR in Turkey.

BACKGROUND: Dirofilaria immitis and Dirofilaria repens are the most common species of filarial nematodes described in the dogs. A single-step multiplex PCR was applied to detect and differentiate simultaneously and unequivocally D. immitis and D. repens on DNA extracted from canine peripheral blood and besides to detect the seroprevalance of D. immitis by ELISA in Elazig Province, Turkey. A PCR detection of the Wolbachia, which plays an important role in D. immitis biology and contributes to the inflammatory pathology of the heartworm, was also applied for the first time in Turkey. METHODS: A total of 161 whole blood and sera samples were collected from stray dogs and stored at -20 °C until used. After DNA extraction, all samples were processed with Dirofilaria primers by multiplex-PCR and Wolbachia primers by conventional PCR besides ELISA for serology. The amplification was performed using a set of primers designed on a portion of the small subunit ribosomal RNA gene of the mitochondrion (12S rDNA). RESULTS: Three of the examined dogs (1.8%) were found to be infected with only D. immitis, one (0.6%) with D. repens and three (1.8%) with both parasites. Besides, 10 out of 161 dogs (6.2%) were found infected with Wolbachia sp. Finaly, the seroprevalence of dirofilariosis in the examined dogs was found to be 3.7% (6/161). CONCLUSION: Although dirofilariosis is not a serious problem in the region, the stray dogs still continue to be a source of infection.

Molecular discrimination of sheep and cattle isolates of Echinococcus granulosus by SSCP and conventional PCR in Turkey.

Cystic echinococcosis (CE), caused by hydatid cysts, is a widespread and hazardous disease in humans and animals worldwide. The aim of the current study was to investigate the genetic characteristics of sheep and cattle isolates of Echinococcus granulosus obtained from eastern Turkey using Single Stranded Conformation Polymorphism (SSCP) analysis and conventional PCR method. A total of 54 isolates collected from Erzurum and Elazig provinces of east-Turkey were examined. The 31 of these were obtained from liver of sheep while 23 cattle isolates (12 of liver and 11 of lung) were tested. After the total genomic DNA isolation 12S rRNA gene of all isolates were examined by PCR for the aim of genetic characterization by conventional PCR and mitochondrial CO1 gene for SSCP analysis. The 12S rRNA-PCR yielded 254 bp of amplification product with all samples analyzed. Thus, these samples were identified as G1-G3 cluster (E. granulosus sensu stricto). At least two major single stranded bands were resolved for G1-G3 cluster and G5 in SSCP analysis. While the resolution of more than two additional single stranded bands in SSCP indicated the existence of G7 genotype. The SSCP analysis was identified the G5 and G7 while failed to G1 and G3. The present SSCP analysis classified all 54 cyst isolates from sheep and cattle as E. granulosus sensu stricto (G1-G3 cluster). However, some sequenced samples for G1 and G3 showed the same band patterns by SSCP.