Mastitisli Sığırlardan İzole Edilen Staphylococcus aureus İzolatlarının Virülens Gen Profillerinin Belirlenmesi

Abstract

Bu çalışmada anabilim dalı kültür koleksiyonunda bulunan subklinik mastitisli sığır sütlerinden izole edilen 140 adet stafilokok izolatından Staphylococcus aureus izolatlarının moleküler identifikasyonu ve bazı önemli virülens genlerinin tespiti amaçlandı. Bu amaçla 140 stafilokok izolatı termonükleaz (nuc) gen varlığı yönünden PZR ile incelendi. nuc geni (279 bp) tespit edilen 42 izolat S. aureus olarak identifiye edildi. Ayrıca izolatlarda lökotoksin geni (pvl), stafilokokal klasik enterotoksin genleri (sea, seb, sec), eksfolyatif toksin genleri (eta, etb), hemolizin genleri (hla, hlb), adezin faktör genleri (fnbA, fnbB, clfa) ve toksik şok sendrom toksin geni (tst) olmak üzere 12 farklı virülens geninin varlığı 3 farklı multipleks PZR kullanılarak araştırıldı. İzolatların 30 (%71.4)’u clfa, 24 (%57.1)’ü hla, 14 (%33.3)’ü hlb, 10 (%23.8)’u fnbB, 6 (%14.2)’sıfnbA, 6 (%14.2)’sıetb, 5 (%11.9)’i sec, 5 (%11.9)’i tst, 3 (%7.1)’ü sea, 2 (%4.7)’si pvl ve 1 (%2.3)’i eta geni açısından pozitif bulundu. İzolatların hiçbirinde seb geni saptanamadı. İzolatların38’inde bir ya da daha fazla virülens geni, 29’unda ise iki ya da daha fazla virülens geni belirlendi. 4 izolatta ise incelenen virülens genlerin hiçbiri saptanamadı. İncelenen izolatlarda en fazla tespit edilen virülens genlerin clfa ve hla olduğu görüldü. Sonuç olarak sınırlı dahi olsa sığır mastitislerinin patogenezinde rol oynayan virülens gen patternleri ortaya konuldu. Çalışma sonuçlarının sığır mastitislerinin etiyolojisinde önemli rol oynayan S. aureus’a ait virülens faktörlerinin karakterizasyon çalışmalarına katkı sağlayacağı kanaatine varıldı.

Keywords

• Mastitis
• Sığır
• S. aureus
• Virülens gen profili

Determination of Virulence Gene Profiles of Staphylococcus aureus Isolated from Cattle with Mastitis

Abstract

This study aimed to determine the molecular identification of Staphylococcus aureus isolates and some important virulence genes from 140 staphylococci samples isolated from cow milk with mastitis in the culture collection of the department. For this purpose, the presence of the thermonuclease (nuc) gene in 140 S. aureus isolates was examined by PCR. Forty‐two isolates that were positive for the nuc gene (279 bp) were identified as S. aureus. In addition, the isolates were examined for the presence of 12 different virulence genes, including leukotoxin gene (pvl), staphylococcal classical enterotoxin genes (sea, seb, sec), exfoliative toxin genes (eta, etb), hemolysin genes (hla, hlb), adhesin factor genes (fnbA, fnbB, clfa), and toxic shock syndrome toxin gene (tst), using three different multiplex PCR. Of the isolates were found to be positive 30 (71.4%) for clfa, 24 (57.1%) hla, 14 (33.3%) hlb, 10 (23.8%) fnbB, 6 (14.2%) fnbA, 6 (14.2%) etb, 5 (11.9%) sec, 5 (11.9%) tst, 3 (7.1%) sea, 2 (4.7%) pvl, and 1 (2.3%) eta genes. The seb gene was not detected in any of the isolates. At the same time one or more virulence genes were determined in 38 of 42 isolates, two or more virulence genes in 29 of the isolates. None of the virulence genes examined in 4 isolates could be detected. It was seen that the most detected virulence genes in the analyzed isolates were clfa and hla. As a result, virulence gene patterns that play a role in the pathogenesis of bovine mastitis, even if limited, were revealed. It was concluded that the results would contribute to the characterization studies of virulence factors belonging to S. aureus, which play an important role in the etiology of bovine mastitis.

Keywords

• Bovine
• Mastitis
• S. aureus
• Virulence gen profile

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