VANKOMİSİN DİRENÇLİ ENTEROCOCCUS FAECALIS İZOLATLARI ÜZERİNDE KATYONİK STEROİD ANTİBİYOTİK CSA-44’ÜN ANTİMİKROBİYAL VE ANTİBİYOFİLM AKTİVİTESİNİN İNCELENMESİ

Abstract

Günümüz kliniklerinde oldukça korkulan bir etken olmaya başlayan Enterococcus faecalis, bu özelliklerini gösterdikleri yüksek antibiyotik direncine borçludur. Ağır antibiyotik stresine maruz kalmalarına rağmen bu direnç özellikleri sayesinde yaşamlarına devam ederek sağaltımı zor enfeksiyonlara neden olmaktadırlar. Sefalosporinler, penisilinler ve trimetoprim/sülfametoksazol gibi oldukça sık ve yaygın kullanılan moleküllere karşı doğal ve kazanılmış direnç gösterebilmesine ek olarak glikopeptitlere karşı ortaya çıkan kazanılmış direnç tedaviyi giderek zorlaşmasına neden olmaktadır. Son seçenek olarak kullanılabilecek vankomisine karşı görülen direncin son yıllarda giderek arttığı rapor edilmiştir. Yaşanan bu krizin aşılmasında antibiyotiklere alternatif olabilecek tüm metotların denenmesi bir zorunluluk haline gelmiştir. Bu çalışmada, yeni katyonik peptid türevi bir molekül olan CSA-44’ün E. faecalis izolatlarına karşı antibakteriyel ve antibiyofilm etkilerinin araştırılması amaçlandı. Bu bakterilerin antimikrobiyal duyarlılıkları otomatize sistemle belirlenerek EUCAST kriterleri doğrultusunda değerlendirildi. Daha sonra seragenin CSA-44 minimum inhibe edici konsantrasyon (MİK) değerleri sıvı mikrodilüsyon metoduyla saptandı. Molekülün zamana bağlı öldürme kinetikleri tespit edildi. CSA-44’ün antibiyofilm aktivitesi kristal viyole metoduyla saptandı. Biyofilme olan etkinlik ayrıca taramalı elektron mikroskobuyla (SEM) görüntülendi. CSA-44’ün MİK değerleri 2-4 µg/mL aralığında saptandı. MİK dozları uygulandığında CSA-44’ün bakteri biyofilmlerini izolata bağlı olarak %50’ye varan oranlarda inhibe ettiği belirlendi. Moleküllerin 2MİK ve MİK konsantrasyonlarında uygulandığında 3-6 saat aralığında bakterisidal etkinlik gösterdiği gözlendi. Ancak, MİK konsantrasyonlarda inkübasyon uzadığında yeniden bakteri miktarında artış olduğu saptandı. Molekülün bakteriyel biyofilmi inhibe ettiği ve membranda hasara neden olduğu SEM ile görüntülendi. Katyonik steroid antibiyotiklerin, dirençli E. faecalis izolatları üzerinde etkili olabileceği bu çalışma tarafından gösterdiğinden, toksikolojik ve farmakodinamik özellikleri inceleyen yeni çalışmaların yapılması uygun olacaktır.

Keywords

• antibiyofilm
• CSA-44
• Enterococcus faecalis

Investigation of the Antimicrobial and Antibiofilm Activity of the Cationic Steroid Antibiotic CSA-44 on Vancomycin-Resistant Enterococcus faecalis Isolates

Abstract

Enterococcus faecalis, which has emerged as a highly feared agent in present day’s clinics, possesses these attributes as a result of its high-level antibiotic resistance. Despite being exposed to significant antibiotic stress, they persist in their existence as a result of these resistance characteristics, leading to infections that are challenging to treat. In addition to the ability to display natural and acquired resistance to common and widely used molecules such as cephalosporins, penicillins, and trimethoprim/sulfamethoxazole, the acquired resistance to glycopeptides makes treatment increasingly challenging. In recent years, there has been a gradual increase in the resistance to vancomycin molecules, which can be employed as a last resort. It has become essential to explore all potential alternatives to antibiotics in order to resolve this crisis. In this study, it was aimed to investigate the antibacterial and antibiofilm effects of CSA-44, a new cationic peptide derivative molecule, against E. faecalis isolates. Antimicrobial susceptibilities of these bacteria were determined with an automated system and evaluated in accordance with EUCAST criteria. Subsequently, minimum inhibitory concentration (MIC) values of ceragenin CSA-44 were determined by broth microdilution method. The time-kill kinetics of the molecule were determined. Antibiofilm activity of CSA-44 was determined by crystal violet method. The antibiofilm activity, was also imaged with scanning electron microscopy (SEM). CSA-44 MIC values were found in the range 2-4 µg/mL. When MIC doses were administered, it was determined that CSA-44 inhibited bacterial biofilms by up to 50% depending on the isolate. The molecules exhibited bactericidal activity during the 3-6-hour period when applied at 2MIC and MIC concentrations. However, at MIC concentrations, the quantity of bacteria increased again as the incubation time prolonged. It was imaged with SEM that the molecule inhibits the bacterial biofilm and causes damage to the membrane. Since this study shows that cationic steroid antibiotics can be effective on resistant E. faecalis isolates, it would be appropriate to conduct new studies examining the toxicological and pharmacodynamic properties.

Keywords

• Enterococcus faecalis
• CSA-44
• antibiofilm

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