Publication: Çevreyi algılama sistemi inhibitörlerinin bazı antibiyotikler ile kombinasyonlarının pseudomonas aeruginosa’ ya karşı antibakteriyel ve antivirülans etkinliklerinin araştırılması
Abstract
Amaç: Çalışmamızda çevreyi algılama (Quorum sensing=QS) sistemi inhibitör maddelerin tek başlarına ve bazı antibiyotiklerle kombinasyonlarının, Pseudomonas aeruginosa’ ya karşı antibakteriyel ve antivirulans etkinlikleri araştırılmıştır.Gereç ve Yöntem: Çalışmada P.aeruginosa PA01, P.aeruginosa 27853 ATCC, P.aeruginosa klinik suş kullanılmıştır. Kullanılan maddelerin minimum inhibitör konsantrasyonları (MİK) sıvı mikrodilüsyon yöntemi ile belirlenmiştir. MİK’leri belirlenen maddelerin P. aeruginosa’nın elastaz aktivitesi üzerine inhibisyon etkileri elastin congo red (ECR), piyosiyanin üretimi üzerini inhibisyon etkileri kloroform ekstraksiyonu yöntemi ile belirlenmiştir. QS sistemi inhibitörlerinin ve antibiyotiklerin biyofilm oluşumuna etkisi kristal viyole metodu kullanılarak araştırılmıştır. İnhibitör maddelerin antibiyotik kombinasyonlarının antimikrobiyal aktivitesi MİK belirleme yöntemi ile belirlenmiştir. Kayma hareketi, kayma testi uygulanarak inokülasyonun yapıldığı noktadan çevreye doğru yayılmanın çapının ölçülmesiyle test edilmiştir.Bulgular: Çalışmada kullanılan kurkumin, piperin, yüksek saflıkta kurkumin, sinnamaldehit, siprofloksasin, meropenem ve tobramisin için MİK değerleri sırasıyla 87,5-175 μg/ mL, 125 μg/ mL, 62,5-125 μg/ mL, 0.0315 μg/ mL, 0,5-0,06 μg/ mL, 0,25-1 μg/ mL, 0,25-2 μg/ mL arasında bulunmuştur. İnhibitör maddelerin P. aeruginosa suşlarının kayma (swarming) hareketi, biyofilm oluşumu, piyosiyanin ve elastaz aktivitesi üzerine inhibisyon etkileri olduğu saptanmıştır. Antibiyotiklerin MİK konsantrasyonunda PA01’ de en etkili tobramisin, klinik suşta da siprofloksasin, MİK (½) konsantrasyonunda ise her iki suşta da siprofloksasin en etkili antibiyotik olarak belirlenmiştir.Sonuçlar: Çalışmada kullanılan QS inhibitör maddelerinin P. aeruginosa üzerinde antibakteriyel ve antivirulans aktiviteleri olduğu saptanmıştır. Antibiyotik ve inhibitör maddelerin kombinasyonlarının tek başlarına etkili oldukları MİK konsantrasyonundan daha düşük konsantrasyonlarda antibakteriyel aktiviteye sahip oldukları saptanmıştır.
Objective: In this study, it was aimed to investigate the effects of natural compounds and combinations with antibiotics on antimicrobial activities and production of P. aeruginosa virulence factors.Material and Methods: İn the study P. aeruginosa PA01, P. aeruginosa 27853 ATCC, P. aeruginosa clinical isolate was used. The MIC of the substances used were determined by the liquid microdilution method. Inhibition effects of substances whose MICs were determined on the elastase activity of P. aeruginosa were determined by elastin congo red (ECR), chloroform extraction method on the production of pyocyanin. The effect of QS system inhibitors and antibiotics on biofilm formation was investigated using crystal violet method. Antimicrobial activity of antibiotic combinations of inhibitors was determined by MIC determination method. The swarming motility was tested by measuring the diameter of the spread from the point of inoculation to the environment by applying the swarming test.Results: MIC values for curcumin, piperine, high purity curcumin, cinnamaldehyde, ciprofloxacin, meropenem and tobramycin used in the study were 87,5-175 μg / mL, 125 μg / mL, 62,5-125 μg / mL, 0,0315 μg / mL respectively It was found between 5-0,06 μg / mL, 0,25-1 μg / mL, 0,25-2 μg / mL. Inhibitors have been found to have inhibition effects on the swarming motility, biofilm formation, pyocyanin and elastase activity of P. aeruginosa strains. In the MIC concentration of antibiotics, the most effective tobramycin in PA01, ciprofloxacin in the clinical strain, and in the MIC (½) concentration, ciprofloxacin was the most effective antibiotic in both strains.Conclusion: The QS inhibitor substances used in the study were found to have antibacterial and antiviral activities on P. aeruginosa. Combinations of antibiotic and inhibitory agents were found to have lower concentrations of antibacterial activity than MIC concentrations that alone were effective.
Objective: In this study, it was aimed to investigate the effects of natural compounds and combinations with antibiotics on antimicrobial activities and production of P. aeruginosa virulence factors.Material and Methods: İn the study P. aeruginosa PA01, P. aeruginosa 27853 ATCC, P. aeruginosa clinical isolate was used. The MIC of the substances used were determined by the liquid microdilution method. Inhibition effects of substances whose MICs were determined on the elastase activity of P. aeruginosa were determined by elastin congo red (ECR), chloroform extraction method on the production of pyocyanin. The effect of QS system inhibitors and antibiotics on biofilm formation was investigated using crystal violet method. Antimicrobial activity of antibiotic combinations of inhibitors was determined by MIC determination method. The swarming motility was tested by measuring the diameter of the spread from the point of inoculation to the environment by applying the swarming test.Results: MIC values for curcumin, piperine, high purity curcumin, cinnamaldehyde, ciprofloxacin, meropenem and tobramycin used in the study were 87,5-175 μg / mL, 125 μg / mL, 62,5-125 μg / mL, 0,0315 μg / mL respectively It was found between 5-0,06 μg / mL, 0,25-1 μg / mL, 0,25-2 μg / mL. Inhibitors have been found to have inhibition effects on the swarming motility, biofilm formation, pyocyanin and elastase activity of P. aeruginosa strains. In the MIC concentration of antibiotics, the most effective tobramycin in PA01, ciprofloxacin in the clinical strain, and in the MIC (½) concentration, ciprofloxacin was the most effective antibiotic in both strains.Conclusion: The QS inhibitor substances used in the study were found to have antibacterial and antiviral activities on P. aeruginosa. Combinations of antibiotic and inhibitory agents were found to have lower concentrations of antibacterial activity than MIC concentrations that alone were effective.