Anaplasma marginale msp1a geninin yeni bir tandem raporunu da içeren, Türkiye’nin Karaman yöresindeki sığırlarda Anaplasma ve Ehrlichia türlerinin moleküler araştırması

Abstract

Kene kaynaklı patojenler evcil ve yabani hayvanlarda ciddi sağlık problemlerine ve verim kaybına neden olur. Karaman ilindeki sığırlarda Anaplasma / Ehrlichia (A/E) ile enfeksiyon sıklığını saptamak için moleküler bir çalışma yapıldı. Venöz kan numuneleri görünüşte sağlıklı olan 150 sığırdan 2016 yılında alınmıştır. A/E türlerinin 16S rRNA geninin değişken V1 bölgesi amplifiye edildikten sonra tür spesifik problar kullanılarak reverse line blot (RLB) deneyi gerçekleştirilmiştir. Bazı örnekler sadece A/E probuna sinyal verdiğinden, örnekler Anaplasma marginale'nin major surface proteinleri (MSPs) açısından analiz edilmiştir. A. marginale'nin msp1α gen dizileri için genetik çeşitlilik ve tandem tekrar analizi yapıldı. Mikroskobik inceleme ile dört (%2.66) hayvanda Anaplasma benzeri cisimler tespit edildi. Anaplasma centrale RLB ile sekiz (%5.33) hayvanda tespit edildi. Örnekler semi-nested PZR ile A. marginale msp1a geni açısından incelediğinde, toplam dokuz (%6.00) hayvanın [6'sı (%4.00) RLB ile A. centrale açısından pozitif olan] A. marginale ile enfekte olduğu bulunmuştur. Ek olarak, MSP1a amplikonlarının dizileri bir tane yeni tandem tekrarı (Tr70) ortaya çıkardı. Bu sonuçlara göre Karaman ilindeki sığırlarda A. marginale ve A. centrale'nin bulunduğu tespit edilmiş ve bu çalışma bölgedeki sığırlarda bir yeni tandem tekrarı ile A. marginale’nin genetik çeşitliliğinin ilk kanıtını sağlamıştır.

Keywords

• A. marginale,A. centrale,sığır,tandem tekrarı,Karaman

Molecular survey of Anaplasma and Ehrlichia species in cattle from Karaman of Turkey, including a novel tandem report of Anaplasma marginale msp1a gene

Abstract

Tick-borne pathogens cause serious health problems and loss of productivity in domesticated and wild animals. A molecular study was performed to detect the frequency of infection with Anaplasma/Ehrlichia (A/E) in cattle from Karaman province of Turkey. Venous blood samples were taken from 150 apparently healthy cattle in 2016. After amplification the hypervariable V1 region of the 16S rRNA gene of A/E species, a reverse line blot (RLB) assay was performed using species-specific probes. Since some samples gave signal only to A/E catch-all probe, the samples analyzed in terms of major surface proteins (MSPs) of Anaplasma marginale. Genetic diversity and tandem repeat analysis were made for msp1α gene sequences of A. marginale. Anaplasma-like bodies were detected in four (2.66%) animals via microscopic examination. Anaplasma centrale was detected in eight (5.33%) animals via RLB. When the samples were examined in terms of A. marginale msp1a gene with semi-nested PCR, a total of nine (6.00%) animals [six of them (4.00%) were positive for A. centrale with RLB] were found to be infected with A. marginale. In addition, the sequences of MSP1a amplicons revealed one new tandem repeat (Tr70). According to these results, it was determined that A. marginale and A. centrale were found in cattle in Karaman province and this study provided the first evidence of genetic diversity of A. marginale with one new tandem repeat in cattle in the region.

Keywords

• A. marginale,A. centrale,cattle,tandem repeat,Karaman

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