Sultan ve Hürmetçi Sazlıklarından Belirli Gıda Patojenlerine Karşı İzole Edilen Litik Fajların Fenotipik Karakterizasyonu

Year 2025, Volume: 6 Issue: 1, 26 - 34, 30.06.2025

Nurhan Ertaş Onmaz , Yasin Özkaya , Rumeysa Nur Karakuş

https://doi.org/10.58833/bozokvetsci.1686329

Abstract

Bu çalışmada, Sultan ve Hürmetçi sazlıklarından spesifik gıda patojenlerine karşı litik bakteriyofajların izole edilmesi, fenotipik ve biyolojik özelliklerinin belirlenmesi ve etkinliklerinin değerlendirilmesi amaçlandı. Bu kapsamda, toplanan su örnekleri, Escherichia coli, Staphylococcus aureus, Salmonella Typhimurium, Listeria monocytogenes ve Pseudomonas aeruginosa bakterilerine özgü litik fajların izolasyonu amacıyla zenginleştirildi; faj varlığı spot test yöntemiyle ön değerlendirildi ve pozitif sonuçlar çift tabaka agar yöntemiyle saflaştırıldı. İzole edilen litik fajların optimal MOI değerleri, adsorpsiyon kinetikleri, tek aşamalı büyüme eğrileri, konak spektrumları ve çevresel stabiliteleri çift tabaka agar yöntemi ve spot test kullanılarak karakterize edildi. Çalışmada, L. monocytogenes hariç diğer test bakterilerine karşı çapları 1-3 mm arasında değişen toplam 40 litik faj izole edildi. Elde edilen fajların titreleri 10⁹-10²⁰ pfu/mL arasında bulundu, tüm faj izolatları 0.1–100 MOI seviyelerinde bakteriyel büyümeyi inhibe etti ve 1 MOI düzeyinde en yüksek çoğalma gözlendi. Fajların adsorpsiyon süreleri, latent periyotları ve lizis süreleri sırasıyla 15-20 dk, 15-25 dk ve 20-30 dk arasında değişirken, patlama boyutlarının 100–120 pfu/cfu arasında olduğu belirlendi. İzole edilen fajlar pH 3.0-9.0 aralığında ve 50°C’ye kadar stabilite gösterdi. Ayrıca, izole edilen fajların 4°C'de altı ay boyunca aktivitelerini koruduğu ve yalnızca hedef bakterilere karşı litik etkinlik göstererek yüksek konak özgüllüğü sergilediği belirlendi. Sonuç olarak, çalışmada sazlıklardan izole edilen fajlar, geniş litik spektrumları, yüksek özgüllükleri ve çevresel koşullara karşı gösterdikleri stabilite özellikleri ile biyokontrol ve terapötik uygulamalarda kullanılabilecek ajan adayları olabilecekleri gösterilmiştir.

Keywords

Bakteriyofaj , Sultan Sazlığı , Hürmetçi Sazlığı , litik etkinlik , stabilite

Phenotypic Characterization of Lytic Phages Targeting Specific Food Pathogens Isolated from Sultan and Hürmetçi Marshes

Abstract

In this study, the isolation of lytic bacteriophages against specific food pathogens from the Sultan and Hürmetçi marshes, the characterization of their phenotypic and biological properties, and the evaluation of their activity were aimed. For this purpose, water samples collected from the marshes were enriched for the isolation of lytic phages specific to Escherichia coli, Staphylococcus aureus, Salmonella Typhimurium, Listeria monocytogenes, and Pseudomonas aeruginosa. The presence of phages was initially assessed by the spot test method, and positive results were subsequently purified using the double-layer agar method. The isolated lytic phages were characterized for their optimal multiplicity of infection (MOI), adsorption kinetics, one-step growth curves, host range, and environmental stability using spot tests and the double-layer agar method. In the study, a total of 40 lytic phages with plaque diameters ranging from 1 to 3 mm were isolated against the test bacteria, excluding L. monocytogenes. The phage titers ranged between 10⁹ and 10²⁰ pfu/mL, and all phage isolates inhibited bacterial growth at MOI levels between 0.1 and 100, with the highest proliferation observed at an MOI of 1. The adsorption times, latent periods, and lysis times of the phages were determined to range between 15-20 min, 15-25 min, and 20–30 min, respectively, while the burst sizes ranged between 100 and 120 pfu/cfu. The isolated phages demonstrated stability across a pH range of 3.0 to 9.0 and up to a temperature of 50°C. Additionally, the phages retained their activity after six months of storage at 4°C and exhibited high host specificity by displaying lytic activity only against their target bacteria. In conclusion, it was shown that phages isolated from reeds in the study could be candidate agents that can be used in biocontrol and therapeutic applications with their broad lytic spectrum, high specificity and stability against environmental conditions.

Keywords

Bacteriophage , Sultan Marshes , Hürmetçi Marshes , lytic activity , stability

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