• Journal of Microbiology and Biotechnology
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• v.26 no.11
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• pp.1829-1835
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• 2016

Dental caries is caused by cariogenic biofilm, an oral biofilm including Streptococcus mutans. Recently, the prevention of dental caries using various probiotics has been attempted. Lactococcus lactis HY 449 is a probiotic bacterium. The aim of this study was to investigate the effect of L. lactis HY 449 on cariogenic biofilm and to analyze its inhibitory mechanisms. Cariogenic biofilm was formed in the presence or absence of L. lactis HY 449 and L. lactis ATCC 19435, and analyzed with a confocal laser microscope. The formation of cariogenic biofilm was reduced in cultures spiked with both L. lactis strains, and L. lactis HY 449 exhibited more inhibitory effects than L. lactis ATCC 19435. In order to analyze and to compare the inhibitory mechanisms, the antibacterial activity of the spent culture medium from both L. lactis strains against S. mutans was investigated, and the expression of glucosyltransferases (gtfs) of S. mutans was then analyzed by real-time RT-PCR. In addition, the sucrose fermentation ability of both L. lactis strains was examined. Both L. lactis strains showed antibacterial activity and inhibited the expression of gtfs, a nd t he d ifference b etween both strains did not show. In the case of sucrose-fermenting ability, L. lactis HY 449 fermented sucrose but L. lactis ATCC 19435 did not. L. lactis HY 449 inhibited the uptake of sucrose and the gtfs expression of S. mutans, whereby the development of cariogenic biofilm may be inhibited. In conclusion, L. lactis HY 449 may be a useful probiotic for the prevention of dental caries.

• 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.

• 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.27 no.9
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• pp.1609-1616
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• 2017

The complex roles of cell surface modification in the biofilm formation of Campylobacter jejuni, a major cause of worldwide foodborne diarrheal disease, are poorly understood. In a screen of mutants from random transposon mutagenesis, an insertional mutation in the eptC gene (cj0256) resulted in a significant decrease in C. jejuni NCTC11168 biofilm formation (<20%) on major food contact surfaces, such as polystyrene and borosilicate glass, when compared with wild-type cells (p < 0.05). In C. jejuni strain 81-176, the protein encoded by eptC modified cell surface structures, such as lipid A, the inner core of lipooligosaccharide, and the flagellar rod protein (FlgG), by attaching phosphoethanolamine. To assess the role of eptC in C. jejuni NCTC11168, adherence and motility tests were performed. In adhesion assays with glass surfaces, the eptC mutant exhibited a $0.77log\;CFU/cm^2$ decrease in adherence compared with wild-type cells during the initial 2 h of the assay (p < 0.05). These results support the hypothesis that the modification of cell surface structures by eptC affects the initial adherence in biofilm formation of C. jejuni NCTC11168. In motility tests, the eptC mutant demonstrated reduced motility when compared with wild-type cells, but wild-type cells with the transposon inserted in a gene irrelevant to biofilm formation (cj1111c) also exhibited decreased motility to a similar extent as the eptC mutant. This suggests that although eptC affects motility, it does not significantly affect biofilm formation. This study demonstrates that eptC is essential for initial adherence, and plays a significant role in the biofilm formation of C. jejuni NCTC11168.

• Restorative Dentistry and Endodontics
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• v.39 no.4
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• pp.288-295
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• 2014

Objectives: This study was designed to evaluate the synergistic antibacterial effect of xylitol and ursolic acid (UA) against oral biofilms in vitro. Materials and Methods: S. mutans UA 159 (wild type), S. mutans KCOM 1207, KCOM 1128 and S. sobrinus ATCC 33478 were used. The susceptibility of S. mutans to UA and xylitol was evaluated using a broth microdilution method. Based on the results, combined susceptibility was evaluated using optimal inhibitory combinations (OIC), optimal bactericidal combinations (OBC), and fractional inhibitory concentrations (FIC). The anti-biofilm activity of xylitol and UA on Streptococcus spp. was evaluated by growing cells in 24-well polystyrene microtiter plates for the biofilm assay. Significant mean differences among experimental groups were determined by Fisher's Least Significant Difference (p < 0.05). Results: The synergistic interactions between xylitol and UA were observed against all tested strains, showing the FICs < 1. The combined treatment of xylitol and UA inhibited the biofilm formation significantly and also prevented pH decline to critical value of 5.5 effectively. The biofilm disassembly was substantially influenced by different age of biofilm when exposed to the combined treatment of xylitol and UA. Comparing to the single strain, relatively higher concentration of xylitol and UA was needed for inhibiting and disassembling biofilm formed by a mixed culture of S. mutans 159 and S. sobrinus 33478. Conclusions: This study demonstrated that xylitol and UA, synergistic inhibitors, can be a potential agent for enhancing the antimicrobial and anti-biofilm efficacy against S. mutans and S. sobrinus in the oral environment.

• Journal of Microbiology and Biotechnology
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• v.32 no.2
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• pp.220-227
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• 2022

The spread of antibiotic-resistant strains of Staphylococcus aureus, a gram-positive opportunistic pathogen, has increased due to the frequent use of antibiotics. Inhibition of the quorum-sensing systems of biofilm-producing strains using plant extracts represents an efficient approach for controlling infections. Torilis japonica is a medicinal herb showing various bioactivities; however, no studies have reported the anti-biofilm effects of T. japonica extracts against drug-resistant S. aureus. In this study, we evaluated the inhibitory effects of T. japonica ethanol extract (TJE) on biofilm production in methicillin-sensitive S. aureus (MSSA) KCTC 1927, methicillin-resistant S. aureus (MRSA) KCCM 40510, and MRSA KCCM 40511. Biofilm assays showed that TJE could inhibit biofilm formation in all strains. Furthermore, the hemolysis of sheep blood was found to be reduced when the strains were treated with TJE. The mRNA expression of agrA, sarA, icaA, hla, and RNAIII was evaluated using reverse transcription-polymerase chain reaction to determine the effect of TJE on the regulation of genes encoding quorum sensing-related virulence factors in MSSA and MRSA. The expression of hla reduced in a concentration-dependent manner upon treatment with TJE. Moreover, the expression levels of other genes were significantly reduced compared to those in the control group. In conclusion, TJE can suppress biofilm formation and virulence factor-related gene expression in MSSA and MRSA strains. The extract may therefore be used to develop treatments for infections caused by antibiotic-resistant S. aureus.

• Journal of Food Hygiene and Safety
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• v.37 no.5
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• pp.328-331
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• 2022

Biofilms are complexly structured communities of microorganisms composed of surface-attached microorganisms, where their effects on the host have been controversial. In this study, we investigated the potential biofilm-forming capacity of Lacticaseibacillus rhamnosus LRH020 (DSM25568) by detecting genes known to promote biofilm formation. It was shown that the aggregation substance gene (asa 1) was presented in the LRH020 strain. Therefore, we investigated the phenotypic activities of the gene asa1 via two methods: biofilm formation and auto-aggregation activity. It was shown that the strain LRH020 had significantly less ability to form biofilm compared to the positive control strain Enterococcus faecalis ATCC 19433. Furthermore, LRH020 exhibited biofilm-forming activity comparable to Lacticaseibacillus rhamnosus GG (LGG), widely used probiotics. The auto-aggregation activity of LRH020 was also within the safe range similar to that of LGG. In conclusion, this study shows that both biofilm-forming and auto-aggregation activities of the LRH020 are comparable to one of the most studied probiotics strains, LGG.

• The Journal of the Convergence on Culture Technology
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• v.8 no.6
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• pp.615-624
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• 2022

In this study, we assessed anti-oxidant activity, anti-microbial, inhibition of biofilm formation and inflammatory factors(nitric oxide, interleukin-6, interleukin-8) inhibitory effect of Perillae semen hydrothermal extract(PSW) and three kinds of Perillae semen supercritical fluid extract(PSSs) extracted by controlling temperature with no variation of pressure. Compared with PSW, PSSs had significantly lower minimal inhibitory concentrations(MICs) against Staphylococcus aureus(S. aureus) and the ability of PSSs to inhibit formation of biofilm was also superior. PSSs reduce the production of inflammatory mediator and inflammatory cytokines significantly compared to PSW. We suggest, therefore, Perillae semen supercritical fluid 45℃ extract which showed the best anti-microbial, inhibition of biofilm formation, and inhibition of inflammatory factors production among the supercritical fluid extracts could be used for protecting patients with atopic dermatitis from pruritus and transepidermal water loss as a functional ingredient from nature.

• Proceedings of the Korean Geotechical Society Conference
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• 2000.03b
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• pp.331-337
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• 2000

When microorganism is injected into porous medium such as soils, biomass retained in the pore. Bacteria within these microcolonies produced large amounts of exopolysaccharides and formed a plugging biofilm. Soil pore size and shape are varied from the initial condition as a result of biofilm formation, which make hydraulic conductivity reduced and friction rate between soil aggregates increased. In this research, hydraulic conductivity reduction was measured after microorganism are inoculated and cultured with synthetic substrate and nutrient. Also, pore sand of before and after biofilm formation compared with scanning electron microscopy. Hydraulic conductivity of Sand and Poorly Graded Sand was decreased approximately 1/10∼1/100 after biomass inoculation and cultivation. Biofilm attached on soil aggregates is resistant to acidic or basic condition.

• Journal of Environmental Science International
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• v.2 no.4
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• pp.331-336
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• 1993

A mathematical model for organic removal efficiency was investigated in a fluidized bed biofilm reactor by changing the feed flow rate, the residence time and the recycle flow rate. In batch experiment, organic removal could be assumed as first order and an intrinsic first order rate constant(k1) was found $6.4{\times}^{-6}cm^3/mg{\cdot}sec$ at influent COD range of 3040 - 6620 mg/L. In continuous experiment, at the condition of the influent COD, 3040 mg/L, the superficial upflow velocity, 0.47 cm/sec, the biofilm thickness 336 ${\mu}m$ and the biofilm dry density 0.091 g/mL, the calculated COD removal efficiency from the mathematical model gave 60% which was very close to the observed value of 66 %. As the feed flow rate was increased, the COD removal efficiency was sharply decreased and at constant feed flow rate, the COD removal efficiency was decreased also as the residence time being decreased.