Investigation on Antimicrobial, Antibiofilm Activities; and Synergistic Effects of Seeds of Two Vincetoxicum Taxa Growing in Turkey

Abstract

Aim: The aim of this study is to examine the antimicrobial, antibiofilm activities, and synergistic effects of the seeds of two Vincetoxicum taxa [Vincetoxicum canescens subsp. pedunculata (VC) and Vincetoxicum fuscatum subsp. fuscatum (VP)] of which some species have been traditionally used to treat neurosis, malaria, scrofula, scabies, internal fever, external cancers, injuries, and wounds.
Methods: Antimicrobial activities of ethanolic extracts obtained from seeds were determined by the microdilution method against 5 reference bacterial strains (Escherichia coli, Pseudomonas aeruginosa, Enterococcus faecalis, Staphylococcus aureus, Klebsiella pneumoniae) and 2 reference fungal strains (Candida albicans and Candida parapsilosis). Effect of the extracts on inhibiting biofilm formation and on formed biofilm was determined by the crystal violet method. Minimum biofilm inhibition concentration (MBIC50) and minimum biofilm reduction concentration (MBRC50) were determined by biofilm tests. The microdilution checkerboard method was used to evaluate the synergistic effect between the two extracts against E.coli.
Results: The extracts inhibited the tested bacteria and yeasts in the minimum inhibition concentrations (MIC) range of 62.5–250 µg/mL. It was determined that tested VF and VC extracts inhibited biofilm formation by 47%, 39%, 50%, and 34% at 0.5X and 0.25X MIC, respectively, and the MBIC50 value of both extracts was 62.5 µg/mL. In addition, it was determined that the extracts inhibited the preformed biofilm by 54%, 62%, and 56%, and 61% at 1X and 2X MIC, respectively, and the MBRC50 value was 125 µg/mL. It was determined that the extracts showed an additive effect (FIC=0.62) against E.coli.
Conclusions: Although the tested extracts have moderate and low antimicrobial effects, they have well effects both in preventing the formation of biofilm and in removing the formed biofilm

Keywords

• Vincetoxicum canescens subsp. pedunculata
• Vincetoxicum fuscatum subsp. fuscatum
• Seed
• Antimicrobial
• Antibiofilm
• Synergy test

Türkiye’de Yetişen İki Vincetoxicum Taksonunun Tohumlarının Antimikrobiyal, Antibiyofilm Aktiviteleri ve Sinerjik Etkilerinin Araştırılması

Abstract

Amaç: Bu çalışmanın amacı bazı türleri geleneksel olarak nevroz, sıtma, skrofula, uyuz, ateş, eksternal kanserler, incinme ve yara tedavisinde kullanılan Vincetoxicum cinsine ait iki taksonun (Vincetoxicum canescens subsp. pedunculata (VC) ve Vincetoxicum fuscatum subsp. fuscatum (VF)) tohumlarının antimikrobiyal, antibiyofilm aktiviteleri ve sinerjik etkilerini incelemektir.
Yöntem: Öğütülmüş tohumlardan elde edilmiş etanollü ekstrelerin antimikrobiyal etkinlikleri mikrodilüsyon yöntemiyle 5 referans bakteri suşu (Escherichia coli, Pseudomonas aeruginosa, Enterococcus faecalis, Staphylococcus aureus, Klebsiella pneumoniae) ve 2 referans maya suşuna (Candida albicans and Candida parapsilosis) karşı test edilmiştir. Ayrıca ekstrelerin P. aeruginosa’nın biyofilm oluşumunu inhibe etme ve oluşmuş biyofilm üzerine etkisi kristal viyole yöntemi ile değerlendirilmiştir. Biyofilm testleri ile minimum biyofilm inhibisyon konsantrasyonu (MBIC50) ve minimum biyofilm azaltma konsantrasyonu (MBRC50) belirlenmiştir. E.coli'ye karşı iki ekstre arasındaki sinerjik etkiyi değerlendirmek için ise mikrodilüsyon dama tahtası yöntemi kullanılmıştır.
Bulgu: Ekstreler, test edilen bakteri ve mayaları 62,5–250 µg/mL minimum inhibisyon konsantrasyonları (MİK) aralığında inhibe etmiştir. VF ve VC ekstrelerinin 0,5X ve 0,25X MİK'de biyofilm oluşumunu sırasıyla %47, %39 ve %50, %34 oranında inhibe ettiği ve iki ekstrenin MBIC50 değerinin 62,5 µg/mL olduğu belirlenmiştir. Ayrıca ekstrelerin önceden oluşmuş biyofilmi 1X ve 2X MİK'de sırasıyla %54, %62 ve %56, %61 oranında inhibe ettiği ve MBRC50 değerinin 125 µg/mL olduğu belirlenmiştir. Dahası ekstrelerin E.coli'ye karşı aditif etki (FİK=0,62) gösterdikleri de tespit edilmiştir.
Sonuç: Test edilen ekstreler orta ve düşük antimikrobiyal etki göstermekle birlikte hem biyofilm oluşumunu engelleme hem de oluşmuş biyofilmi ortadan kaldırmada iyi etkinliğe sahiptir.

Keywords

• Vincetoxicum canescens subsp. pedunculata
• Vincetoxicum fuscatum subsp. fuscatum
• Tohum
• Antimikrobiyal
• Antibiyofilm
• Sinerji testi

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