Molecular survey of Anaplasma phagocytophilum in small ruminants from Şanlıurfa

Yıl 2016, Cilt 6, Sayı 2, 1 - 8, 01.12.2016

Müzeyyen ATAŞ Nazir Dumanlı Kürşat ALTAY

Öz

This study was performed to determine the presence and prevalence of Anaplasma phagocytophilum in sheep and goats in the Şanlıurfa region. Whole blood samples were taken from a total of 208 animals (134 sheep and 74 goats) in the Şanlıurfa region. Polymerase chain reaction (PCR) using primers specific to the A. phagocytophilum 16S SSU rRNA gene of the bacterium was applied to the blood samples. The amplified DNA fragments in PCR positive samples were sequenced in order to confirm their identity. PCR amplification indicated the presence of A. phagocytophilum in 9 animals (4.33 %). A. phagocytophilum was present in 5.97 % (8/134) of sheep and 1.35 % (1/74) of goats. The nucleotide sequence of a sample selected to represent the PCR positive samples was determined. The resulting DNA sequence was compared to other DNA sequences available in the GenBank using BLAST analysis and placed into GenBank under accession number as KJ183079. The presence of A. phagocytophilum in the sheep and goats in the Şanlıurfa region was demonstrated for the first time using PCR and sequence analyses and epidemiologic data regarding the pathogen were also obtained.

Anahtar Kelimeler

Anaplasma phagocytophilum, goat, PCR, sheep, Şanlıurfa

Şanlıurfa yöresinde koyun ve keçilerde Anaplasma phagocytophilum’un moleküler yöntemlerle araştırılması

Öz

Bu araştırma, Şanlıurfa yöresinde koyun ve keçilerde Anaplasma phagocytophilum’un varlığı ve yaygınlığının belirlenmesi amacıyla yapılmıştır. Bu amaçla, Şanlıurfa yöresinde 134 koyun ve 74 keçi olmak üzere toplam 208 hayvandan tam kan örneği alınmıştır. Bu örneklerden total genomic DNA’lar izole edilmiş, elde edilen DNA örnekleri Anaplasma phagocytophilum’un varlığı yönünden 16S SSU rRNA genine spesifik primerler kullanılarak, polimeraz zincir reaksiyonu (PZR)’na tabi tutulmuştur. Polimeraz zincir reaksiyonu sonucunda pozitif olduğu belirlenen örnekleri temsilen seçilen bir örneğin 16S SSU rRNA geninin DNA dizilimi belirlenmiştir. İncelenen 208 koyun ve keçiye ait kan örneğinin 9 (% 4.33)’unda A. phagocytophilum’un varlığını gösteren PZR amplifikasyon ürünü elde edilmiştir. A. phagocytophilum pozitiflik oranının koyunlarda % 5.97 (8/134), keçilerde % 1.35 (1/74) olduğu saptanmıştır. Pozitif örnekleri temsilen seçilen bir örneğinin DNA dizilimi belirlenmiş, bu dizilim BLAST analiz yoluyla GenBankasındaki diğer dizilimlerle karşılaştırıldıktan sonra GenBanka KJ183079 accession numarası ile kayıt edilmiştir. Bu çalışma ile Şanlıurfa yöresindeki koyun ve keçilerde PZR ve sekans analizi ile A. phagocytohilium varlığı ilk defa ortaya konmuş ve etkene ait epidemiyolojik veriler de ilk olarak elde edilmiştir

Anahtar Kelimeler

Anaplasma phagocytophilum, koyun, keçi, PZR, Şanlıurfa

Kaynakça

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