The Effect of Nisin and Chloramphenicol Combinationon Staphylococcus aureus ATCC 6538 Biofilm Structure

Abstract

Nowadays, due to the rapid spread of antibiotic resistance and the difficulty of discovering new antimicrobial agents, the reuse and combinational strategies of old antibiotics have come to the fore. In this study, we investigated the combinational efficacy of nisin and chloramphenicol against the ATCC 6538 strain of Staphylococcus aureus. The MIC values of nisin and chloramphenicol were >64 and 32 µg/mL, respectively. The checkerboard assay was carried out for the inspection of synergism between nisin and chloramphenicol. Furthermore, the crystal violet assay was employed to assess antibiofilm effects. Additionally, the expressions of various virulence genes (agrA, spa, icaA and saeR) were investigated using the colony biofilm assay and qRT-PCR methods. In combination application, MIC values of nisin and chloramphenicol was decreased. In addition, biofilm formation was decreased.It was shown that the expression level of the agrA gene decreased compared to the control in the nisin, chloramphenicol and combinational applications. The expression levels of other genes were increased compared to the control. The results showed that the activities of nisin and chloramphenicol combinations had synergistic and antibiofilm activity. This study sheds light on the combinatorial use of older antibiotics.

Keywords

• Staphylococcus aureus
• Combination
• Antibiotic resistance
• Nisin
• Chloramphenicol

Nisin ve Kloramfenikol Kombinasyonunun Staphylococcus aureus ATCC 6538 Biyofilmi Üzerindeki Etkileri

Abstract

Günümüzde antibiyotik direncinin hızla yayılması ve yeni antimikrobiyal ajanların keşfedilmesinin zorluğu nedeniyle eski antibiyotiklerin yeniden kullanımı ve kombinasyon stratejileri ön plana çıkmıştır. Bu çalışmada, Staphylococcus aureus'un ATCC 6538 suşuna karşı nisin ve kloramfenikolün kombinasyonel etkinliğini araştırdık. Nisin ve kloramfenikolün MİK değerleri sırasıyla >64 ve 32 µg/mL olarak bulundu. Nisin ve kloramfenikol arasındaki sinerjizmin incelenmesi için dama tahtası (Checkerboard) testi yapıldı. Ayrıca, antibiyofilm etkilerini değerlendirmek için kristal viole testi kullanıldı. Koloni biyofilm testi ve qRT-PCR yöntemleri kullanılarak çeşitli virülans genlerinin (agrA,spa, icaA ve saeR) ekspresyonları araştırıldı. Kombinasyonel uygulamada, nisin ve kloramfenikolün MİK değerlerinde azalma gözlendi. Ayrıca, kombinasyonel uygulamada, biyofilm oluşumu azalma gösterdi. Nisin, kloramfenikol ve kombinasyon uygulamalarında agrA geninin ekspresyon seviyesinin kontrole göre azaldığı gösterilmiştir. Diğer genlerin ekspresyon seviyeleri ise kontrole göre artış göstermiştir. Sonuçlarımız, nisin ve kloramfenikol kombinasyonlarının aktivitelerinin sinerjistik ve antibiyofilm aktiviteye sahip olduğunu göstermiştir. Bu çalışma, eski antibiyotiklerin kombinasyonel kullanımının önemine ışık tutmaktadır.

Keywords

• Staphylococcus aureus
• Kombinasyon
• Antibiyotik dirençliliği
• Nisin
• Kloramfenikol

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