Vibrio vulnificus Klinik İzolatlarının Virülens Genlerine Dayalı Olarak Selektif Zenginleştirme Ortamının Geliştirilmesi

Abstract

Bu çalışmanın amacı, farklı zenginleştirme besiyerlerinin V. vulnificus'un klinik ve çevresel suşlarının seçimini etkileyip etkilemediğini bulmaktı. Bu nedenle, geleneksel zenginleştirme ve koruyucu besiyerlerinin selektif özellikleri, V. vulnificus’ un klinik ve çevresel suşları için test edildi. Laboratuarda, aynı tuz konsantrasyonuna sahip Alkali Peptonlu Su (APW) ve Beyin Kalp İnfüzyon Broth (BHIB) eşit konsantrasyonlarda klinik ve çevresel suşlar ile ekimi yapıldı ve 35 °C' de 20 saat boyunca inkübe edildi. Kantitatif Polimeraz Zincir Reaksiyon (PZR) sayım sonuçları; BHIB’ de geliştirilen suşların %85’ ini klinik suşlar olduğunu göstermiştir. Bu oran vcgC geninin vvhA genine yüzdelik hesaplamalarıyla belirlenmiştir. APW' de klinik suşların (47%) çevresel suşlara (53%) oranı neredeyse eşitti. BHIB, APW ve Luria Bertani (LB) broth ile zenginleştirilen deniz suyu numunelerinin büyük kısmında, klinik suşlarının toplam suşlara oranı BHIB’de yüksek çıkarken, APW ve LB’de çıkmamıştır. Ancak, deniz suyu örneklerinde klinik veya çevresel suşların birbirine oranında mevsimsel farklılıklar zenginleşme sürecini etkilemiştir. Çalışmamız BHIB10'un virülans faktörlerini çalışmak için avantajlı olabileceğini ve APW10'un V. vulnificus'ta izolasyon ve MPN sayımı için uygun olabileceğini göstermektedir. Tuzluluk oranı %10 olan BHIB besiyerinin klinik suşlar için selektif özelliği, demirin insanlarda oluşturduğu infeksiyonlarda önemli bir rol oynadığını gösterir.

Keywords

• Vibrio vulnificus,Selektif zenginleştirme,vcgC,vvhA

Development of Selective Enrichment Medium for Clinical Isolates of Vibrio vulnificus Based upon Virulence Correlating Genes

Abstract

The objective of this study was to find out if different enrichment media would influence the selection of clinical and environmental strains of Vibrio vulnificus. Therefore, traditional enrichment and maintenance medium for V. vulnificus were tested for selective preference for clinical or environmental strains. In the laboratory, Alkaline Peptone Water (APW) and Brain Heart Infusion Broth (BHIB), with adjusted salinity to that of APW, were inoculated with equal concentrations of clinical and environmental strains and incubated for 20 hr at 35 °C. BHIB selected for clinical strains (85%) as indicated by percentage of vcgC to vvhA genes, enumerated with quantitative PCR. In APW, the ratio of clinical (47%) to environmental strains (53%) was roughly equal. Enrichments of seawater with BHIB, APW, and Luria Bertani broth (LB) resulted in higher percentage of clinical strains in BHIB but not in APW or LB, in most samples. However seasonal differences in predominance of clinical or environmental strains in the seawater samples influenced the enrichment process. Our study suggests that BHIB10 might be advantageous for studying virulence factors and APW10 could be suitable for isolation and MPN enumeration in V. vulnificus. The selective preference of the BHIB10 medium for clinical strains indicates that iron plays an important role in the infection of the human host.

Keywords

• Vibrio vulnificus,Selective enrichment,vcgC,vvhA

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