Abstract

Bu çalışma, süt ürünleri proses hatlarından izole edilen hem Gram-negatif hem de biyofilm üreticisi bakterilerin Quorum Sensing (QS) sinyallerini göstermeyi amaçlamıştır. QS sinyallerinin profilinin ve kimyasal bileşiminin tanımlanması, mikrobiyal direnç gelişimi ve biyofilm oluşumunun kontrolü için önemli bir faktördür. Özellikle Gram-pozitif ve Gram-negatif izolatların olağan dışı davranışlarına odaklanılmıştır. İzolatların uzun zincirli açil-homoserin lakton (AHL) sinyalleri (C14-HSL, C16-HSL ve C18-HSL) ve DFD (4,5-dihidroksi-2,3-pentanedion)-AI-2 sinyalleri yüksek performanslı sıvı kromatografisi (HPLC) ve Tandem kütle spektrometresi ile Sıvı Kromatografisi (LC-MSMS) yöntemleri ile incelenmiştir. Sonuç olarak, tüm Gram-pozitif izolatların olağandışı olarak AHL üreticisi olduğu tespit edilmiştir. Biyosensör ve HPLC yöntemleri ile AHL üreticisi olmadığı tespit edilen Gram-negatif bakteriler, LC-MSMS yöntemi ile AHL üreticileri olarak tanımlandı. Gram-pozitif izolatlar tarafından üretilen DFD sinyali, Gram-negatif izolatlardan Klebsiella pneumonia, Enterobacter cloacae ve Klebsiella oxytoca'da tespit edilmiştir. Sonuçlar, QS sisteminin karmaşık bir sistem olduğunu ve biyosensör mikroorganizmaların QS sinyal tanımlaması için en iyi yöntem olmayabileceğini göstermiştir. Bu çalışma, mikroorganizmaların biyofilm oluşumunu azaltmak ve direnç kazanımını önlemek için kimyasal iletişimi tamamen kesmek adına QS sinyallerinin profilini ve kimyasal bileşimini tanımlama konusunda önemli bir bakış açısı sunmaktadır.

EVALUATION OF QUORUM SENSING SIGNALS OF STRONG BIOFILM PRODUCING BACTERIA VIA LC-MSMS, HPLC AND BIOSENSORS

Abstract

This study aimed to show presence of Quorum Sensing (QS) signals of Gram-negative and Gram-positive biofilm producing bacteria isolated from real dairy process lines. Defining the profile and chemical composition of QS-signals is an important factor in control of microbial resistance and biofilm production. We especially focused on unusual behaviour of Gram-positive and Gram-negative isolates. Long-chain acyl-homoserine lactones (AHLs) signals (C14-HSL, C16-HSL and C18-HSL) and DFD (4,5-dihidroksi-2,3-pentanedione)-AI-2 signals of the isolates were studied by High-performance liquid chromatography (HPLC) and Liquid Chromatography with tandem mass spectrometry (LC-MSMS) methods. All Gram-positive isolates were defined as AHL-producers. All Gram-negative isolates, formerly defined as non-AHL producers by both biosensors and HPLC methods, were identified as AHL-producers. DFD signal was only detected from Gram-negative Klebsiella pneumonia, Enterobacter cloacae and Klebsiella oxytoca isolates. The results demonstrated that the QS-system is a complex system and biosensor microorganism may not be the best method for QS-signal identification. The results also provided new insights in defining the profile and chemical composition of QS-signals importance for interrupting the chemical communication completely to reduce biofilm formation and prevent resistance gain of microorganisms.

Keywords

• AHL Signals
• AI-2 Signals
• Biofilm
• Biosensors
• HPLC
• LC-MSMS

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