IADR Abstract Archives
Lactobacillus-reuteri supplemented Glycerol Reduces Enterococcus-faecalis and Candida-albicans Biofilm Growth
Objectives: Probiotic L. reuteri is known to produce antibacterial substances called reuterin in presence of glycerol. Objective of this study is to analyze L. reuteri Indonesian strain supplemented glycerol effect towards Enterococcus-faecalis and Candida-albicans biofilm growth.
Methods: L.reuteri collected from 20 Indonesian subjects (18-24years), grown in MRS-agar, 37°C, anaerobically. Single colonies were further cultured separately in broth medium before DNA extraction and confirmed with PCR, DNA sequencing and BLAST. Each E.faecalis ATCC-29212 and C.albicans ATCC-10231 cultured in BHI-broth, then distributed into 96well-microplate and incubated for 24-hours, 37°C to form biofilm. L.reuteri clinical isolate was cultured in MRS broth, 24h, anaerobic condition and suspended with glycerol (300mM) for 3-hours. Subsequently, diluted into different concentrations (108,106,and 104CFU/mL) and distributed into biofilm-containing well with incubation for 1h,3h,6h, and 24h. NaOCl (5,25%) and nystatin (100.000IU) used as positive-control. Biofilm wells without probiotic used as negative-control. Biofilm measured using crystal-violet dye using microplate-spectrophotometer (490nm). Data statistically analysed by one-way ANOVA test, p<0.05 set as level of significance.
Results: PCR result showed 1100bp confirmed presence of L.reuteri. BLAST result showed two L. reuteri strains obtained from Indonesian isolates had 96% and 95% values compared to L.reuteri ATCC-55730. It confirmed presence of different strains according to Average Nucleotide Identity(ANI). Biofilm assay result showed significant reduction of biofilm growth of E.faecalis and C.albicans after treatment with L. reuteri supplemented-glycerol in all concentrations and time periods (p<0.05). The ideal concentration to inhibit E. faecalis and C.albicans biofilm was L. reuteri 108 CFU/mL in 1 hour incubation period for both species with 83.36% and 56.41% of biofilm reduction respectively compared to control (p<0.05) set at 100%.
Conclusions: L.reuteri Indonesian strain supplemented glycerol reduces the biofilm growth of E.faecalis and C.albicans. This anti-biofilm activity may be useful in preventing biofilm growth in root canal. Further studies are needed to explore this result.
Methods: L.reuteri collected from 20 Indonesian subjects (18-24years), grown in MRS-agar, 37°C, anaerobically. Single colonies were further cultured separately in broth medium before DNA extraction and confirmed with PCR, DNA sequencing and BLAST. Each E.faecalis ATCC-29212 and C.albicans ATCC-10231 cultured in BHI-broth, then distributed into 96well-microplate and incubated for 24-hours, 37°C to form biofilm. L.reuteri clinical isolate was cultured in MRS broth, 24h, anaerobic condition and suspended with glycerol (300mM) for 3-hours. Subsequently, diluted into different concentrations (108,106,and 104CFU/mL) and distributed into biofilm-containing well with incubation for 1h,3h,6h, and 24h. NaOCl (5,25%) and nystatin (100.000IU) used as positive-control. Biofilm wells without probiotic used as negative-control. Biofilm measured using crystal-violet dye using microplate-spectrophotometer (490nm). Data statistically analysed by one-way ANOVA test, p<0.05 set as level of significance.
Results: PCR result showed 1100bp confirmed presence of L.reuteri. BLAST result showed two L. reuteri strains obtained from Indonesian isolates had 96% and 95% values compared to L.reuteri ATCC-55730. It confirmed presence of different strains according to Average Nucleotide Identity(ANI). Biofilm assay result showed significant reduction of biofilm growth of E.faecalis and C.albicans after treatment with L. reuteri supplemented-glycerol in all concentrations and time periods (p<0.05). The ideal concentration to inhibit E. faecalis and C.albicans biofilm was L. reuteri 108 CFU/mL in 1 hour incubation period for both species with 83.36% and 56.41% of biofilm reduction respectively compared to control (p<0.05) set at 100%.
Conclusions: L.reuteri Indonesian strain supplemented glycerol reduces the biofilm growth of E.faecalis and C.albicans. This anti-biofilm activity may be useful in preventing biofilm growth in root canal. Further studies are needed to explore this result.