Vibrio anguillarum’u Enfekte Eden vY6 ve vY28 Bakteriyofajlarının Izolasyonu ve Ön Karakterizasyonu

Öz

Çoklu ilaca dirençli Vibrio türleri, hem halk sağlığı hem de küresel akuakültür endüstrisi için önemli bir tehdit oluşturmaktadır. Bu türler arasında yer alan Vibrio anguillarum, yetiştiriciliği yapılan balıklarda görülen vibriozis salgınlarının başlıca etkenlerinden biri olarak kabul edilmektedir. Bu çalışmada, V. anguillarum’u enfekte eden iki yeni bakteriyofaj (vY6 ve vY28), Türkiye’deki bir akuakültür çiftliğinden alınan su örneklerinden izole edilmiş ve biyolojik kontrol ajanı olma potansiyellerini değerlendirmek amacıyla karakterize edilmiştir. Tek basamaklı büyüme eğrisi analizi sonucunda, latent periyotlar vY6 için 20 dakika, vY28 için ise 25 dakika olarak belirlenmiştir. Her iki fajın da geniş bir pH ve sıcaklık aralığında stabil kaldığı gözlemlenmiştir. Tüm genom dizileme çalışmaları; vY6 için 84.553 bp, vY28 için ise 99.873 bp büyüklüğünde taslak genomları ortaya çıkarmış ve bu genomlarda sırasıyla 57 ve 55 tahmini kodlayıcı dizi saptanmıştır. Yapılan analizlerde, her iki genomda da antimikrobiyal direnç genine veya virülansla ilişkili bir gene rastlanmamıştır. Genom anotasyonu aracılığıyla; yapısal bileşenlerin sentezi, DNA paketlenmesi, replikasyon ve konak hücre lizisi ile ilişkili genler tanımlanmıştır. V. anguillarum’a karşı antibakteriyel aktivite, 600 nm dalga boyunda optik yoğunluk (OD600) değişimlerinin izlenmesiyle değerlendirilmiştir. Faj uygulaması sonucunda, bakteriyel büyümenin kontrol grubuna kıyasla anlamlı şekilde baskılandığı tespit edilmiştir. Deneyin 24. saati sonunda vY6 bakteriyel büyümeyi yaklaşık %61, vY28 ise %48 oranında azaltırken; faj kombinasyonunun yaklaşık %69’luk bir inhibisyon sağladığı görülmüştür. Canlı hücre sayımı analizleri de bu güçlü antibakteriyel aktiviteyi doğrulamış ve enfeksiyonun erken evrelerinde 5.62 ± 0.71 log birimlik bir azalma kaydedilmiştir. Elde edilen bulgular, vibriozisin kontrolüne yönelik faj temelli yaklaşımların geliştirilmesi için güçlü bir temel oluşturmaktadır.

Anahtar Kelimeler

• akuakültür
• antimikrobiyal direnç
• faj terapi
• vibriozis

Isolation and Preliminary Characterization of Bacteriophages vY6 and vY28 Infecting Vibrio anguillarum

Öz

Multi-drug resistant Vibrio species pose a significant threat to both public health and the global aquaculture industry. Among them, Vibrio anguillarum is one of the major causative agents of vibriosis outbreaks in farmed fish. In this study, two novel bacteriophages (vY6 and vY28) infecting V. anguillarum were isolated from water samples collected at an aquaculture farm in Türkiye and characterized to evaluate their potential as biological control agents. One-step growth curve analysis showed latent periods of 20 min for vY6 and 25 min for vY28 and both phages remained stable across a wide range of pH and temperature conditions. Whole genome sequencing revealed draft genome sizes of 84.553 bp for vY6 and 99.873 bp for vY28, encoding 57 and 55 predicted coding sequences, respectively, with no detectable antimicrobial resistance genes or virulence-associated genes. Genome annotation identified genes involved in structural assembly, DNA packaging, replication, and host cell lysis. Antibacterial activity against V. anguillarum was evaluated by monitoring optical density at 600 nm. Phage treatment significantly suppressed bacterial growth compared to the untreated control. After 24 h, vY6 and vY28 reduced bacterial growth by approximately 61% and 48%, respectively, while the phage combination resulted in approximately 69% inhibition. Viable cell count analysis confirmed strong antibacterial activity, with reductions of up to 5.62 ± 0.71 log during the early stages of infection. These findings provide a basis for further studies exploring phage-based approaches to control vibriosis.

Anahtar Kelimeler

• aquaculture
• antimicrobial resistance
• phage therapy
• vibriosis

Kaynakça

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