Typing of Mannheimia haemolytica isolates isolated from respiratory tract and investigation of virulence genes

Öz

In this study, it was aimed to determine the biochemical properties, distribution of important virulence genes of Mannheimia haemolytica isolates identified from the respiratory tracts of sick and healthy cattles and sheeps. 48 (87.3%) Mannheimia haemolytica isolates from naso-pharyngeal and trachea-bronchial swaps were identified as Mannheimia haemolytica by Real Time-PCR. According to the differences in arginine and sorbitol tests, 4 different biochemical profiles were determined in the isolates examined. Three virulence gene profiles were detected in the isolates examined by Real-Time PCR. 37.5%, 33.3%, 12.5% of the isolates examined were identified as I, III and II, respectively. While all virulence-related genes were identified in the isolates with virulence gene profile II, it was determined that there were no nmaA gene in profile I isolates and nmaA and tbpB genes in profile III isolates. At the same time, it was determined that biochemical profile II was associated with disease cases and this was related to arginine negativity. In addition, it was determined that isolates with virulence gene profile I were associated only with biochemical profile I and that this was due to arginine negativity, whereas the relationship between arginine positive isolates and virulence gene profile III was found to be significant. As a result; arginine negativity and gcp, gs60, tbpB, lktC, adh positive, nmaA negative isolates may be the epidemiological criteria that can be used to differentiate commensal and pathogen Mannheimia haemolytica isolates and new studies on the subject should be done.

Anahtar Kelimeler

• Mannheimia haemolytica
• Typing
• Virulence genes

Solunum Sisteminden İzole Edilen Mannheimia haemolytica izolatlarının tiplendirilmesi ve virulens genlerinin araştırılması

Öz

Bu çalışmada, hasta/sağlıklı sığır ve koyunların solunum yollarından izole edilen Mannheimia haemolytica izolatlarının biyokimyasal özellikleri, önemli virülens genlerinin dağılımı tespit edilmesi amaçlandı. Nazo-farengiyal ve trake-bronşiyal svap örneklerinden Real-Time PCR ile 48 (%87.3) adet Mannheimia haemolytica izolatı kullanıldı. İncelenen izolatlarda, hastalık ve virülens genleri ile ilişkili biyokimyasal özelliklerin arginin ve sorbitol testlerindeki farklılıklara göre 4 farklı biyokimyasal profil belirlendi. Real Time-PCR yöntemiyle virülens genleri incelenen izolatlarda, %37.5’i I, %33.3’ü III ve %12.5’i II olmak üzere 3 farklı virülens gen profili tespit edildi. Virülens gen profil II’ye sahip izolatlarda virülens ile ilişkili genlerin tamamı belirlenirken, profil I’e sahip izolatlarda nmaA, profil III’e sahip izolatlarda ise nmaA ve tbpB genlerinin bulunmadığı tespit edildi. Bununla birlikte biyokimyasal profil II’nin hastalık olguları ile ilişkili ve bunun arginin negatiflik ile ilişkili olduğu belirlendi. Ayrıca virülens gen profil I özelliğine sahip izolatların sadece biyokimyasal profil I ile ilişkili olduğu ve bunun arginin negatiflikten kaynaklandığı belirlenirken, arginin pozitif izolatlar ile virülens gen profil III arasındaki ilişkinin önemli olduğu gözlendi. Sonuç olarak; arginin negatiflik ile gcp, gs60, tbpB, lktC, adh pozitif, nmaA negatif izolatların kommensal ve patojen Mannheimia haemolytica izolatlarının ayrımında kullanılabilecek epidemiyolojik kriter olabileceği ve konu ile ilgili yeni çalışmaların yapılması gerektiği düşünülmektedir.

Anahtar Kelimeler

• Mannheimia haemolytica
• Tiplendirme
• Virülens gen

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