Koyun ve Sığırlardan İzole Edilen Arcanobacterium pyogenes Suşlarının Moleküler Karakterizasyonu

The purpose of this study was to determine the presence of virulence genes, perform typing for the characterization of Arcanobacterium pyogenes field strains, and investigate the correlation between clonal types, virulence genes and occurence of disease. The isolates (n=51) used in this study were isolated from different sources (liver (n=4) of sheep; liver (n=25), lungs (n=5), broncho-alveolar lavage (BAL) fluid (n=3), milk (n=12) and suppurative tissue (n=1) of dairy cows and synovial fluid (n=1) of a calf ). The presence of haemolytic activity in A. pyogenes isolates was determined using rabbit, sheep, cattle, chicken, dog and human erythrocytes. Also, the presence of any cytotoxic effect was investigated by growth in Vero cell cultures. Genomic DNA fingerprinting for clonal analysis was generated by BOX-PCR typing. Conventional PCR was used for the determination of the presence of eight A. pyogenes virulence factor genes, namely, nanH (encoding neuraminidase H), nanP (encoding neuraminidase P), plo (encoding pyolysin-PLO), cbpA (encoding collagen-binding protein A) and fimA, fimC, fimE and fimG (encoding the major fimbrial subunit of four different fimbriae). Furthermore, the correlation between clonal types, virulence factors and the occurence of disease was investigated. The haemolysins of all strains had haemolytic effect on rabbit, sheep, cattle, chicken, dog and human erythrocytes. In addition, all strains were found to be cytotoxic to Vero cells. According to clonal analysis results, the A. pyogenes isolates were determined to belong to 12 different types. While all A. pyogenes strains were positive for the plo gene, the positivity rate was 62% for the nanH gene, 84% for the nanP gene, 58% for the cbp gene, 96% for the fimA gene, 66% for the fimC gene, 42% for the fimE gene and 10% for the fimG gene. It was determined that no correlation existed between the clonal types and virulence factors of A. pyogenes isolates and occurence of disease.