Faham KHAMESIPOUR, Abbas DOOSTI, Mohammad SEBDANI MAZROUEI

Survey for the presence of Mycobacterium avium subsp. paratuberculosis in the bull frozen semen samples and blood samples of cattle, sheep and camel by nested-PCR

Mycobacterium avium subsp. paratuberculosis (MAP) Mycobacterium cinsi içinde zorunlu, Gram pozitif, aside dirençli bir bakteridir. Bu bazı hayvanlarda Johnes hastalığı ve belki de insanlardaki Crohns hastalığı etkenidir. Bu çalışmada nested-PCR ile sığır, deve ve koyunların herbir örneğinde MAPın belirlenmesi amaçlanmıştır. Kan numuneleri 144 sığır, 110 koyun ve 95 deveden toplandı ve 83 dondurulmuş semen örneği boğalardan elde edildi, takiben DNA izolasyonu gerçekleştirildi. Bu sırada, nested-PCR MAPın IS900 geni için özel primerler ile gerçekleştirildi. İkiyüz otuz bp uzunluğuna sahip PCR ürünleri, pozitif olarak tahmin edildi. İranda suni tohumlamada kullanılan boğalardan elde edilen 83 semen örneğinin 8inde (%9.638) ve sığır, deve ve koyunlardan alınan sırasıyla 144te 11 (%7.638), 95te 7 (%7.368) ve 110da 16 (%14.545) kan örneğinde MAP belirlendi. Bu çalışmanın sonuçları, nested-PCRın sığır, deve ve koyunlarda MAP gibi hücre içi bakterilerin belirlenmesinde yüksek verimlilikli iyi bir teknik olduğunu göstermektedir. Bu sonuçlar sığır, deve ve koyun Johnes hastalığında MAPin hızlı teşhisi için daha fazla dikkatin gerekli olduğunu ortaya koymaktadır.

Mycobacterium avium subsp. paratuberculosis varlığının dondurulmuş boğa semen örneklerinde ve sığır, koyun ve deve kan örneklerinde nested-PCR ile taranması

Mycobacterium avium subsp. paratuberculosis (MAP) is an obligate, Gram positive, acid-fast bacterium in the genus Mycobacterium . It is the causative agent of Johnes disease in some animals and perhaps the human disease Crohns disease. The aim of this study was to use nested- PCR as an exact and fast technique to trace MAP in cattle, camel and sheep from each samples. Blood samples were collected from 144 cattle, 110 sheep and 95 camels and 83 frozen semen samples were obtained from bulls were obtained and DNA was extracted. At that time, nested-PCR was performed by specific primers for IS900 gene of MAP. The PCR products with 230 bp length were estimated as a positive. The occurrence of MAP detected in 8 of the 83 (9.638%) frozen semen samples of bulls that used for artificial insemination, and was detected in 11 of the 144 (7.638%), 7 of the 95 (7.368%) and 16 of the 110 (14.545%) blood samples of cattle, camel and sheep in Iran, respectively. The results of this study were displayed nested-PCR is a good technique with high efficiency for detection of intracellular bacteria such as MAP in cattle, camel and sheep samples. Consequently, more attention to Johnes disease in cattle, camel and sheep to find MAP quickly is essential.

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