Biofilms associated with human infection have high levels of pathogenicity due to their resistance to antibiotics. The discovery of an active antibiofilm agent against polymicrobial biofilms is a necessary consequence for coping with biofilmrelated infections. Thymol and Eugenol are essential oils that have potential as antibacterial and antifungal. This study aimed to determine the effectiveness of thymol and eugenol inhibits C. albicans, P. Aeruginosa, E. coli S. aureus and polymicrobial biofilm. Biofilm formation inhibition assay and biofilm degradation assay of thymol and eugenol were determined using microtiter broth method.The antibiofilm efficacy of thymol and eugenol towards polymicrobial biofilms were analyzed by calculating minimum biofilm inhibitor concentration (MBIC50) and minimum biofilm eradication concentration (MBEC50) values. The data were analyzed using Statistical Package for the Social Sciences (SPSS) with 95% confidence level. Thymol and eugenol showed inhibitory activity against the formation of mono and polymicrobial biofilms of the microbial tested.The result also demonstrated an evidence of activity of thymol and eugenol in breaking down mono and polymicrobial biofilm. Therefore, thymol and eugenol serves as a potential source for new antibiofilm drugs towards polymicrobial biofilm.
Background:As part of our search for new potential natural resources to eradicate infection, we have revealed the prominent potency of massoia bark (Massoia aromatica Becc, Lauraceae) in combating immunosuppressed-related infection.Materials and Methods:The extract was prepared by macerating the pulverized dried bark in ethanol 95%, followed by solvent evaporation. The oil was extracted from the dried bark by steam-hydrodistillation of which preparative thin-layer chromatography was performed on the oil to isolate the active constituent, C-10 massoia lactone (ML). Anti-biofilm assay against Candida albicans was conducted on polystyrene 96 wells microtiter plates, followed by a confocal laser scanning microscope observation to get three-dimensional profiles of the affected biofilms. Effects on the hyphae development inoculated on RPMI-1640 agar plates were observed for 7 days. Influences of samples on mice macrophage phagocytosis were examined by an in vitro technique. Samples concentration tested were in the range of 2.0–0.0625 mg/mL and done in triplicate.Results:Massoia bark extracts (oil and solid phase) and ML exhibited promising activities as anti-biofilm against C. albicans at IC50 0.074% v/v, 271 μg/mL and 0.026 μg/mL, respectively. The ML did not inhibit the hyphae development at the concentration tested; however, the extracts showed inhibition at 62.5 μg/mL. Macrophage phagocytosis stimulation was correlated to the ML content.Conclusion:Massoia bark is potential to be developed as anti-infective in immunosuppressed condition of which the C10 ML (C10H16O2) plays a major role in exerting activity.SUMMARY Massoia bark extracts (oily and solid phase) and C-10 Massoia lactone exhibited promising activities as antibiofilm against Candida albicans at IC50 are 0.074 %v/v, 271 μg/mL and 0.026 μg/mL respectively. The major constituent, C-10 Massoia lactone (C10H16O2) plays major role in exerting anticandida activity and potentially acts as an immunomodulator as well. However extracts showed inhibition of hyphae development of C. albicans which showed no correlation to the content of the Massoia lactone. Abbreviations used: GC/MS: Gas Chromatography/Mass Spectrometry, ML: Massoia Lactone, TLC: Thin Layer Chromatography, ATCC: American Type Culture Collection, RPMI: Roswell Park Memorial Institute, PBS: Phosphate Buffer Sterile, LSM: Laser scanning microscope, DMSO: Dimethyl sulfoxide, UV: Ultra violet, SDB: Sabouraud dextrose agar, MeOH: Methanol, LB: Luria Bertani, EtOAc: Ethyl acetate, CLSM: Confocal Laser Scanning Microscope, PI: Propidium iodide.
The essential oil of Massoia (Massoia aromatica Becc., Lauraceae) bark is a potential immunomodulator in vitro. This study evaluated the potential immunomodulatory effects of Massoia bark infusion on the nonspecific immune response (phagocytosis) of Wistar rats. For the in vitro assay, macrophages were treated with the freeze-dried infusion at the concentrations of 2.5, 5, 10, 20, or 40 µg/mL media. For the in vivo assay, two-month-old male Wistar rats were divided into five groups. The baseline group received distilled water at the dose of 1 mL/100 g body weight (BW), with the herbal product containing Phyllanthus niruri extract that was administered as the positive control at the dose of 0.54 mL/rat. The treatment groups received the infusion at a dose of 100, 300, or 500 mg/100 g BW. Treatments were given orally every day for 14 days. The ability of macrophage cells to phagocyte latex was determined as phagocytic index (PI), and it was observed under microscopy with 300 macrophages. The in vitro study revealed that the phagocytic activity of the infusion-treated macrophages significantly increased in comparison with that of the control macrophages in a concentration-dependent manner. Among all of the treatment concentrations, the concentration of 40 µg/mL provided the highest activity with a PI value of 70.51 ± 1.11%. The results of the in vivo assay confirmed those of the in vitro assay. The results of the present study indicate that Massoia bark can increase the phagocytic activity of rat macrophage cells.
Biofilm acts as the mediator for infection nowadays. Approximately, more than 80% infection incidents are biofilm-formation related. Biofilm as bacteria's defense system is more difficult to eradicate by antibiotic; therefore, pathogen bacteria on their biofilm forms can make serious problems for human health. The invention of a new candidate for polymicrobial biofilm can be an essential challenge to be studied, in order to prevent infections related to biofilm. Tannin is a polyphenol compound with anti-bacterial and anti-fungal potential. This study aims to acknowledge the effectiveness of tannin in inhibition and degradation of C. albicans, P. aeruginosa, E. coli, S. aureus, and polymicrobial biofilm. The assay for biofilm inhibition and degradation were determined with microtiter broth method. The effectivity of tannin antibiofilm against polymicrobial biofilm were analyzed by calculating minimum biofilm inhibitory concentration (MBIC50) and minimum biofilm eradication concentration (MBEC50) values. The mechanism of action of tannin against polymicrobial biofilm was tested using scanning electron microscopy (SEM). The data were analyzed using the Statistical Package for the Social Sciences (SPSS) with a 95% confidence level. Tannin 1% gave inhibition activity of mono-species biofilm formation S. aureus in the middle phase and maturation of 79.04±0.01, 61.48±0.03, E. coli 74.56±0.01, 67.91±0.02, P. aeruginosa 67.32±0.05, 35.13± 0.01, C. albicans 60.62±0.01, 47.16±0.01. The results also provide evidence that tannin activity can degrade and damage the matrix of extracellular polymeric substance (EPS) polymicrobial biofilms. Hence, tannins can be a potential candidate for new antibiofilm for polymicrobial biofilm.
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