Abstract
The rapid rise of resistance causes existing antibiotics to become dysfunctional. Therefore, search for new antimicrobial active ingredients has increased in recent years. In this study, flower extracts of Castanea sativa were examined for antimicrobial and anti-quorum sensing aspects. The antimicrobial properties of methanol, ethyl acetate, ethanol and hexane extracts of C. sativa against some gram-positive and gram-negative bacterial species, as well as yeasts (Candida albicans and Candida parapsilosis) were investigated by the agar well diffusion method. The minimum inhibition concentration (MIC) and minimum bactericidal concentrations (MBC) of C. sativa extracts were also determined. Chromobacterium violaceum ATCC 12472, C. violaceum 35352, C. violaceum VIR07 and C. violaceum CV026 indicator strains were used for determination of the quorum sensing inhibitions, and the Pseudomonas aeruginosa PAO1 strain was used for the swarming tests. Additionally, biofilm inhibition was detected by the spectrophotometric method using the P. aeruginosa PAO1 strain. Methanol, ethanol and ethyl acetate extracts of C. sativa was found to have high antibacterial and antifungal effects, while the methanol extract also had anti-quorum sensing, anti- swarming and biofilm inhibition effects, but no activity was found in the n- hexane extract. To the best of our knowledge this is the first report revealed that methanol extract obtained from C. sativa flowers induced anti-quorum sensing activities mainly inhibited the violacein production, swarming and biofilm formation. The present investigation provided evidence that the C. sativa flower extract maybe a potential source of antimicrobial agents. Therefore, much attention should be paid to C. sativa flower content, which could be used with high efficacy against microorganisms.
Highlights
The effectiveness of current antibiotics on microorganisms has been decreasing rapidly in recent years
The results showed that the chestnut membrane had more antimicrobial compounds namely flavanol, glycosideand terpenoid than its leaf and shell parts
This study investigated the effects of methanol, ethyl acetate, ethanol and hexane extracts obtained from the flowers of chestnut plant for the antimicrobial, anti-quorum sensing and biofilm inhibition properties
Summary
Collection of C. sativa plant and preparation of its extracts The flowers of C. sativa used in the study were collected from İkizdere (Rize, Turkey), cleaned and fresh flowers were stored at -20 °C until used. The extracts were obtained using methanol, ethyl acetate, ethanol and hexane by maceration method (Solanki and Nagori, 2012). 10-20 g of the flower of chestnut stored at. 68 -20 °C were weighed, ground into fine powder and mixed with 100 or 200 mL solvent. Extraction was conducted on a shaker at room temperature (RT) for 48 h. The extracts were filtered through filter paper and the solvents were evaporated in an evaporator The extracts were suspended at 50-200 mg/mL in Dimethyl sulfoxide (DMSO) and aliquots of these were stored at -20 °C until needed for further assays. Pre-experimental working concentrations were diluted to 10 mg/mL, and the concentrations to be used in the experiments were adjusted from these dilutions
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