Laurus nobilis Esansiyel Yağının Antibakteriyel Potansiyelinin ve 1,8-Sineol ile Gentamisin Arasındaki Sinerjik Etkileşimin Metisiline Dirençli Staphylococcus aureus’a Karşı İncelenmesi

Öz

Bu çalışmada, ticari olarak satın alınan Laurus nobilis uçucu yağının (LnEO) metisilin-dirençli S. aureus üzerine in vitro antibakteriyel aktivitesi ve 1,8-sineol’ün gentamisin ile sinejistik etkileşiminin belirlenmesi amaçlanmıştır. LnEO, GC-MS ile karakterize edilmiş, in vitro antibakteriyel aktivitesi ise disk difüzyon metodu ile test edilmiştir. MIC ve MBC değerleri broth makro-dilüsyon yöntemi ile değerlendirilmiştir. LnEO’nın S.aureus suşunu öldürme kinetiği araştırılmıştır. 1,8-sineolün gentamisin ile etkileşimi dama tahtası yöntemiyle değerlendirilmiştir. GC-MS analizi ile toplam uçucu yağın %100'ünü oluşturan 45 uçucu bileşik belirlenmiştir. Ana bileşenin ise 1,8-cineole (%51,43) olduğu gösterilmiştir. LnEO, 1:40 ve 1:20 seyreltme oranında S. aureus’un gelişimi üzerinde herhangi bir etki göstermemiştir. LnEO, 1:1 seyreltme oranında 14,2 mm, 1:5 seyreltme oranında 11,8 mm ve 1:10 seyreltme oranında ise 8,7 mm inhibisyon zonu oluşturmuştur. S. aureus’un 1:1, 1:5 ve 1:10 seyreltme oranlarında LnEO’na duyarlı olduğu sonucuna varılmıştır. MIC ve MBC değerleri sırasıyla 1:4 ve 1:2 olarak belirlenmiştir. Öldürme zamanı deneyinde, LnEO’nın bakterisit aktiviteye sahip olduğunu gösterilmiştir. LnEO ile 10 dakika temas süresi sonunda S. aureus’un %95’inin öldüğü, 90 dakika temas sonunda ise bakteri suşunun tamamının öldüğü tespit edilmiştir. Gentamisin ile 1,8-sineol kombinasyonu S. aureus üzerinde sinerjik etki göstermiştir (FICI≤ 0.5). Elde edilen veriler, LnEO’nın ve bileşiminde bulunan 1,8-sineol’ün MRSA enfeksiyonlarıyla mücadelede etkili stratejiler geliştirilmesine yardımcı olabileceğini göstermektedir. İlaç etken maddeleri yüksek dozda tek başına kullanılmak yerine, doğal biyoaktif bileşenlerle daha düşük dozda kombine edilerek kullanıldığında olası toksik etkiyi ve direnç gelişimini azaltmasının mümkün olabileceği düşünülmektedir.

Anahtar Kelimeler

• Metisilin dirençli Staphylococcus aureus
• Laurus nobilis
• 1,8 sineol
• Antibakteriyal aktivite
• Gentamisin

Exploring the Antibacterial Potential of Laurus nobilis Essential Oil and the Synergistic Interaction with 1,8-Cineole and Gentamicin Against Methicillin-Resistant Staphylococcus aureus

Abstract

This study aims to determine the in vitro antibacterial activity of the commercially purchased Laurus nobilis essential oil (LnEO) and the synergistic interaction of the 1,8-cineole with gentamicin against methicillin-resistant S. aureus. The chemical profile of the LnEO was evaluated by GC-MS. In vitro antibacterial activity of the LnEO was tested by the disk diffusion method. MIC and MBC values were assessed using the broth macro-dilution method. The killing kinetics of the LnEO against S. aureus were investigated. The interaction of the 1,8-cineole with gentamicin was assessed by the checkerboard method. 45 volatile compounds constituting 100% of the total essential oil were determined. 1,8-cineol (51.43%) was shown as the main component. The LnEO does not affect the growth of S. aureus at 1:40 and 1:20 dilution ratios. The LnEO formed an inhibition zone of 14.2 mm at a 1:1 dilution ratio, 11.8 mm at 1:5, and 8.7 mm at 1:10. It was concluded that S. aureus is sensitive to the LnEO at dilution ratios of 1:1, 1:5, and 1:10. The MIC and the MBC values were determined as 1:4, and 1:2, respectively. Time-kill assay has shown that the LnEO has a bactericidal activity. It was determined that 95% of S. aureus died after 10 minutes of contact with the LnEO, and after 90 minutes of contact, the entire bacterial strain was killed. The combination of gentamicin and 1,8-cineole showed a synergistic effect on S. aureus (FICI≤ 0.5). Obtained data show that LnEO and the 1,8-cineole contained in its composition may help develop effective strategies to combat MRSA infections. It is thought that it may be possible to reduce possible toxic effects and resistance development when drug-active ingredients are used in combination with natural bioactive components at lower doses instead of being used alone at high doses.

Keywords

• Methicillin resistant Staphylococcus aureus
• Laurus nobilis
• 1,8 cineole
• Antibacterial activity
• Gentamicin

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