• Korean Journal of Food Science and Technology
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• v.38 no.1
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• pp.135-139
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• 2006

Most bacteria form biofilm as self-defence system, making efficient food sanitization, preservation, and instrument washing more difficult. Biofilm formation of Salmonella, E. coli, B. cereus, and S. aureus was observed during 24 hr food preservations by performing microtiter plate and glass wool assays. Most cells formed biofilm and attached onto glass wool. When biofilm formation and injury were analyzed on the microtiter plate, 10 and 20% acid-injured E. coli and S. aureus, respectively, 30-50% cold temperature $(4^{\circ}C)-injured$ B. cereus and E. coli, and 30-55% 6% sodium chloride solution-injured Salmonella showed significant biofilm formation. Results indicate biofilm formation level differed within species depending on type of stress.

• Journal of Microbiology and Biotechnology
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• v.27 no.7
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• pp.1242-1248
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• 2017

Several medicinal plants are ethnomedically used in Korea as agents for treating infection, anti-inflammation, and pain relief. However, beyond typical inhibitory effects on cell growth, little is known about the potential anti-biofilm activity of these herbs, which may help to prevent cavities and maintain good oral health. This study aimed to investigate the antimicrobial and anti-biofilm activities of the methanol extracts of 37 Korean medicinal plants against dental pathogens Streptococcus mutans and Candida albicans, which synergize their virulence so as to induce the formation of plaque biofilms in the oral cavity. The antimicrobial activities were investigated by broth dilution and disk diffusion assay. The anti-biofilm and antioxidant activities were evaluated based on the inhibitory effect against glucosyltransferase (GTase) and the DPPH assay, respectively. Among 37 herbs, eight plant extracts presented growth and biofilm inhibitory activities against both etiologic bacteria. Among them, the methanol extracts (1.0 mg/ml) from Camellia japonica and Thuja orientalis significantly inhibited the growth of both bacteria by over 76% and over 83% in liquid media, respectively. Minimum inhibitory concentration (MIC) values of these methanol extracts were determined to be 0.5 mg/ml using a disk diffusion assay on solid agar media. Biofilm formation was inhibited by more than 92.4% and 98.0%, respectively, using the same concentration of each extract. The present results demonstrate that the medicinal plants C. japonica and T. orientalis are potentially useful as antimicrobial and anti-biofilm agents in preventing dental diseases.

• Journal of the Korea Society of Computer and Information
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• v.21 no.5
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• pp.135-139
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• 2016

Scrophularia ningpoensis hemsl has been traditionally used in China and Vietnam for treatment of bacteria, atopy, pimple, tonsillitis, angina and encephalitis for a long time. The main objectives of this study were to evaluate the antibacterial activity of the Scrophularia ningpoensis hemsl extract on biofilm formation of Klebsiella pneumoniae. Antibacterial activity was conducted using disc diffusion assay and minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) were determined using the broth micro dilution method in accordance to Clinical and Laboratory Standards Institute guidelines(CLSI). Furthermore, cytotoxicity on L929 were assessed using animal cell culture for the proliferation test(MTT cell assay) and the biofilm forming capacity of the K. pneumoniae were determined using the colony forming unit (CFU) assay. The extract exhibited considerable antibacterial activity. K. pneumoniae was susceptible to the extract with the MIC and MBC of 0.1875 and $1.5mg/m{\ell}$ respectively. Cytoxicity test in L929 showed no sign of toxicity at the concentration of $0.75mg/m{\ell}$ and at the same concentration the extract caused inhibition of bacterial biofilm formation. The extract of Scrophularia ningpoensis hemsl possesses an in vitro antibacterial antibiofilm activities against K. pneumoniae, with no sign of cytoxicity on L929.

• Journal of Environmental Science International
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• v.30 no.5
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• pp.369-378
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• 2021

In this study, we investigated the effects of indole on biofilm formation inhibition in Pantoea agglomerans (P. agglomerans). In the biofilm growth assay, indole inhibited biofilm formation across all the growth time. Depending on biofilm growth stage, indole exhibited biofilm inhibition and anti-bacterial effects on planktonic cells. Through the analysis of the proportion rate between biofilm and Colony Forming Units (CFU) and inhibition rate of indole, we confirmed that depending on the biofilm stage of P. agglomerans, indole treatment timing was more important than the treatment duration. By comparing gene expression rates through rt-qPCR P.agglomerans affected by indole was found to significantly change quorum sensing (pagI/R) and indole transportation (bssS) gene expressions. Throughout all, indole exhibited both antimicrobial and anti-biofilm effects on P. agglomerans. In addition, we confirmed the anti-biofilm effects of indole on mature biofilm. In conclusion, indole as a signal molecule, can exhibit anti-biofilm effects through bacterial quorum sensing inhibition and indole affects. Therefore, indole can regulate biofilm bacteria especially gram-negative opportunistic pathogens.

• Journal of Microbiology and Biotechnology
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• v.33 no.6
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• pp.715-723
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• 2023

Microbial biofilms are resilient, immune-evasive, often antibiotic-resistant health challenges, and increasingly the target for research into novel therapeutic strategies. We evaluated the effects of a nutraceutical enzyme and botanical blend (NEBB) on established biofilm. Five microbial strains with known implications in chronic human illnesses were tested: Candida albicans, Staphylococcus aureus, Staphylococcus simulans (coagulase-negative, penicillin-resistant), Borrelia burgdorferi, and Pseudomonas aeruginosa. The strains were allowed to form biofilm in vitro. Biofilm cultures were treated with NEBB containing enzymes targeted at lipids, proteins, and sugars, also containing the mucolytic compound N-acetyl cysteine, along with antimicrobial extracts from cranberry, berberine, rosemary, and peppermint. The post-treatment biofilm mass was evaluated by crystal-violet staining, and metabolic activity was measured using the MTT assay. Average biofilm mass and metabolic activity for NEBB-treated biofilms were compared to the average of untreated control cultures. Treatment of established biofilm with NEBB resulted in biofilm-disruption, involving significant reductions in biofilm mass and metabolic activity for Candida and both Staphylococcus species. For B. burgdorferi, we observed reduced biofilm mass, but the remaining residual biofilm showed a mild increase in metabolic activity, suggesting a shift from metabolically quiescent, treatment-resistant persister forms of B. burgdorferi to a more active form, potentially more recognizable by the host immune system. For P. aeruginosa, low doses of NEBB significantly reduced biofilm mass and metabolic activity while higher doses of NEBB increased biofilm mass and metabolic activity. The results suggest that targeted nutraceutical support may help disrupt biofilm communities, offering new facets for integrative combinational treatment strategies.

• Journal of the Korean Wood Science and Technology
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• v.47 no.2
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• pp.178-188
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• 2019

Licorice, which has an extensive history of use as an herbal medicine, has been suggested to have oral health benefits. However, to date, no systematic study has been conducted on the preparation method of licorice extracts for oral health. In this study, licorice extracts prepared using water and ethanol were investigated for its ability to inhibit the biofilm formation of Streptococcus mutans. The licorice extract prepared with around 60% ethanol effectively inhibited the biofilm formation of S. mutans. Licorice extracted with 50% ethanol almost completely inhibited the biofilm formation at 1.5 g/L of licorice extract. This inhibitory activity was confirmed in a microplate assay and a flow cell system. Glycyrrhetic acid was extracted from licorice effectively with 60% ethanol concentration. The strong inhibitory activity of glycyrrhetic acid and the synergistic inhibition with glycyrrhizin on biofilm formation were suggested as major reasons for a concentration-specific extraction. These results suggest that licorice extract prepared using around 60% ethanol effectively inhibits the biofilm formation of S. mutans.

• Restorative Dentistry and Endodontics
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• v.38 no.2
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• pp.65-72
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• 2013

Objectives: To evaluate the inhibitory effect of ursolic acid (UA)-containing composites on Streptococcus mutans (S. mutans) biofilm. Materials and Methods: Composite resins with five different concentrations (0.04, 0.1, 0.2, 0.5, and 1.0 wt%) of UA (U6753, Sigma Aldrich) were prepared, and their flexural strengths were measured according to ISO 4049. To evaluate the effect of carbohydrate source on biofilm formation, either glucose or sucrose was used as a nutrient source, and to investigate the effect of saliva treatment, the specimen were treated with either unstimulated whole saliva or phosphate-buffered saline (PBS). For biofilm assay, composite disks were transferred to S. mutans suspension and incubated for 24 hr. Afterwards, the specimens were rinsed with PBS and sonicated. The colony forming units (CFU) of the disrupted biofilm cultures were enumerated. For growth inhibition test, the composites were placed on a polystyrene well cluster, and S. mutans suspension was inoculated. The optical density at 600 nm ($OD_{600}$) was recorded by Infinite F200 pro apparatus (TECAN). One-way ANOVA and two-way ANOVA followed by Bonferroni correction were used for the data analyses. Results: The flexural strength values did not show significant difference at any concentration (p > 0.01). In biofilm assay, the CFU score decreased as the concentration of UA increased. The influence of saliva pretreatment was conflicting. The sucrose groups exhibited higher CFU score than glucose group (p < 0.05). In bacterial growth inhibition test, all experimental groups containing UA resulted in complete inhibition. Conclusions: Within the limitations of the experiments, UA included in the composite showed inhibitory effect on S. mutans biofilm formation and growth.

• Journal of Microbiology and Biotechnology
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• v.28 no.1
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• pp.77-86
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• 2018

The human pathogen and food spoiler Bacillus cereus can form biofilms that act as a persistent source of contamination, which is of public health concern. This study aimed to understand how the source of isolation might affect the behavior of biofilm formation. Biofilm formation abilities of 56 strains of B. cereus isolated from different environments, including human food poisoning, farm, and food, were determined. Crystal violet assay results revealed significant (p < 0.05) differences in biofilm formation abilities among the strains isolated from different sources only at an early stage of incubation. However, strain origin showed no impact on later stage of biofilm formation. Next, correlation of the group of isolates on the basis of their biofilm-forming abilities with the number of sessile cells, sporulation, and extracellular polymeric substance (EPS) formation was determined. The number of sessile cells and spores in biofilms was greatly influenced by the groups of isolates that formed dense, moderate, and weak biofilms. The contribution of extracellular DNA and/or proteins to EPS formation was also positively correlated with biofilm formation abilities. Our results that the source of isolation had significant impact on biofilm formation might provide important information to develop strategies to control B. cereus biofilm formation.

• Journal of Microbiology and Biotechnology
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• v.25 no.11
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• pp.1871-1879
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• 2015

The complex roles of flagella in the pathogenesis of Campylobacter jejuni, a major cause of worldwide foodborne diarrheal disease, are important. Compared with the wild-type, an insertional mutation of the flgA gene (cj0769c) demonstrated significant decrease in the biofilm formation of C. jejuni NCTC11168 on major food contact surfaces, such as polystyrene, stainless steel, and borosilicate glass. The flgA mutant was completely devoid of flagella and non-motile whereas the wild-type displayed the full-length flagella and motility. In addition, the biofilm formation of the wild-type was inversely dependent on the viscosity of the media. These results support that flagellar-mediated motility plays a significant role in the biofilm formation of C. jejuni NCTC11168. Moreover, our adhesion assay suggests that it plays an important role during biofilm maturation after initial attachment. Furthermore, C. jejuni NCTC11168 wild-type formed biofilm with a net-like structure of extracellular fiber-like material, but such a structure was significantly reduced in the biofilm of the flgA mutant. It supports that the extracellular fiber-like material may play a significant role in the biofilm formation of C. jejuni. This study demonstrated that flgA is essential for flagellar biosynthesis and motility, and plays a significant role in the biofilm formation of C. jejuni NCTC11168.

• Journal of Korean Society of Water and Wastewater
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• v.19 no.2
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• pp.181-192
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• 2005

This study evaluated the effect of factors influencing the initial biofilm formation in drinking water distribution pipe by running experiments using a $2^{4-1}$ fractional factorial experimental design with a replicate. Important variables used for assessing biofilm formation included BDOC(biodegradable dissolved organic carbon), viable heterotrophic bacteria present in drinking water, water temperature, and shear stress at two levels each. Based on the statistical analysis of biofilm levels measured as attached HPC(heterotrophic plate count) and community-level assay, the main factors that have significant effects on biofilm formation were found to be viable heterotrophic bacteria and BDOC. Water temperature only exhibited significant effect on the levels of attached HPC, while shear stress was not a significant factor under given conditions. Moreover, the statistical analysis revealed that interactions between the important variables were not statistically significant at a 0.05 significance level.