Detection of dirofilaria immitis and dirofilaria repens by multiplex-pcr, investigation of wolbachia dna by pcr and determination of dirofilaria immitis seroprevalance in stray dogs of Elazig province

Dirofilaria immitis and Dirofilaria repens are the most common species of filarial nematodes described in the dogs. In this research we aimed to performe a single-step multiplex PCR to detect and differentiate simultaneously and unequivocally D.immitis and D.repens on DNA extracted from canine peripheral blood and also detect the seroprevalance of D.immitis by ELISA. 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). We also aimed to PCR detection of the Wolbachia which is play an important role in D.immitis biology and contributes to the inflammatory pathology of the heartworm. For this aim, 161 blood and sera samples were collected from stray dogs and stored at -20 °C until used. After the DNA extraction, all samples will be worked with Dirofilaria primers by multiplex-PCR and Wolbachia primers by conventional PCR besides ELISA for serology. In this study, three of examined dogs (1.8 % ) were found to be infected with only D.immitis, one of them (0.6 % ) with D.repens and three of them (1.8 % ) with both parasites. Besides, 10 out of 161 (6.2 % ) were detected as infected with Wolbachia sp. Lastly, the seroprevalence of dirofilariosis in examined dogs were found to be 3.7 % (6/161). The simultaneous detection of both D. immitis and D. repens has been achived for the investigation of Wolbachia bacteria in those samples were made first time in our region. As a result, it was concluded that although dirofilariosis is not a serious problem in the region, stray dogs still continue to be a source of infection.