• Korean Journal of Fisheries and Aquatic Sciences
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• v.57 no.2
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• pp.129-136
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• 2024

Lactic acid bacteria (LAB) growth in processed meat products produces slime. In this study, 10 different biofilm-forming LAB, including Leuconostoc mesenteroides, Lacticaseibacillus paracasei, Levilactobacillus brevis, Lactiplantibacillus plantarum, Leuconostoc citreum, Weissella viridescens, and Latilactobacillus sakei, were isolated from various meat products and identified based on 16S rRNA gene analysis. To inhibit biofilm formation by LABs, Eisenia bicycles methanolic extract (EB) and ethyl acetate soluble fraction (EA) were used as antibacterial and antibiofilm agents, respectively. Furthermore, EA and EB were employed to synthesize gold nanoparticles (AuNPs) such as EB-AuNPs and EA-AuNPs, which could serve as antibiofilm agents against the isolated LAB. These findings demonstrate that EA, EB-AuNPs, and EA-AuNPs exhibit significant antibacterial activity against the isolated LAB. Furthermore, EB-AuNPs reduced L. citreum biofilm production, whereas EA-AuNPs inhibited L. mesenteroides and L. brevis biofilm formation. The current results suggest that EB-AuNPs and EA-AuNPs can be used as nanomaterials to inhibit LAB that form biofilms on meat products.

• Journal of Life Science
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• v.33 no.11
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• pp.936-943
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• 2023

This study was conducted to investigate the potential of Stewartia koreana as anti-microbial materials. The branches, stems and leaves of S. koreana were extracted into 70% ethanol and their antibacterial activity against P. aeruginosa was confirmed. The leaf, branch and stems extracts (1 mg/disc) showed the antibacterial activity against P. aeruginosa and leaf extracts showed higher antibacterial activities than those from branch extracts. The MIC against P. aeruginosa was 0.8 mg/ml and showed bacteriostatic action. The inhibitory effects of extract on biofilm formation and gene expression related to biofilm formation of P. aeruginosa was determined by biofilm biomass staining, SEM and qRT-PCR analysis. The biofilm biomass and cell growth of P. aeruginosa in the cultures treated with 0.2~2.0 mg/ml of S. koreana leaf extracts were significantly decreased in a concentration-dependent manner. We observed that the extract had an inhibitory effect on the formation of P. aeruginosa biofilms at concentrations of 0.8 mg/ml by SEM. qRT-PCR analysis showed that the lasI and rh1I gene expression associated to quorum sensing (QS) in the cultures treated with 0.2~2.0 mg/ml of S. koreana leaf extracts were suppressed in a concentration-dependent manner. Based on the above results, it can be concluded that S. koreana leaf extracts can be used as anti-microbial material derived from natural materials, as demonstrated by the antibacterial action and inhibition of biofilm formation of P. aeruginosa by QS inhibition.

• Journal of Applied Biological Chemistry
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• v.64 no.4
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• pp.357-362
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• 2021

In this study, to investigate the anticariogenic effect of red ginseng, the antibacterial effect, cell adherence and biofilm formation inhibitory effect of n-hexane extract of red ginseng marc (HERGM) against Streptococcus mutans, the causative bacteria of caries, were measured. The growth of S. mutans was inhibited in proportion to the concentration of HERGM, and was hardly observed at a concentration above 125 ㎍/mL (MIC =125 ㎍/mL). It was found that HERGM acts on the cell membrane and the nucleic acid component of the cell was leaked. In addition, HERGM inhibited the adherence and biofilm formation of S. mutans by more than 90% at a concentration of 125 ㎍/mL. GTase activity was completely inhibited at a concentration of 50 ㎍/mL of HERGM. In conclusion, it was found that HERGM commonly inhibited the growth and biofilm formation of S. mutans.

• Restorative Dentistry and Endodontics
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• v.46 no.2
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• pp.20.1-20.11
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• 2021

Objectives: The aim of this study was to evaluate bacterial nanocellulose (BNC) membranes incorporated with antimicrobial agents regarding cytotoxicity in fibroblasts of the periodontal ligament (PDLF), antimicrobial activity, and inhibition of multispecies biofilm formation. Materials and Methods: The tested BNC membranes were BNC + 1% clindamycin (BNC/CLI); BNC + 0.12% chlorhexidine (BNC/CHX); BNC + nitric oxide (BNC/NO); and conventional BNC (BNC; control). After PDLF culture, the BNC membranes were positioned in the wells and maintained for 24 hours. Cell viability was then evaluated using the MTS calorimetric test. Antimicrobial activity against Enterococcus faecalis, Actinomyces naeslundii, and Streptococcus sanguinis (S. sanguinis) was evaluated using the agar diffusion test. To assess the antibiofilm activity, BNC membranes were exposed for 24 hours to the mixed culture. After sonicating the BNC membranes to remove the remaining biofilm and plating the suspension on agar, the number of colony-forming units (CFU)/mL was determined. Data were analyzed by 1-way analysis of variance and the Tukey, Kruskal-Wallis, and Dunn tests (α = 5%). Results: PDLF metabolic activity after contact with BNC/CHX, BNC/CLI, and BNC/NO was 35%, 61% and 97%, respectively, compared to BNC. BNC/NO showed biocompatibility similar to that of BNC (p = 0.78). BNC/CLI showed the largest inhibition halos, and was superior to the other BNC membranes against S. sanguinis (p < 0.05). The experimental BNC membranes inhibited biofilm formation, with about a 3-fold log CFU reduction compared to BNC (p < 0.05). Conclusions: BNC/NO showed excellent biocompatibility and inhibited multispecies biofilm formation, similarly to BNC/CLI and BNC/CHX.

• Journal of Microbiology and Biotechnology
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• v.24 no.9
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• pp.1216-1225
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• 2014

Biofilm-related infections of Candida albicans are a frequent cause of morbidity and mortality in hospitalized patients, especially those with immunocompromised status. Options of the antifungal drugs available for successful treatment of drug-resistant biofilms are very few, and as such, new strategies need to be explored against them. The aim of this study was to evaluate the efficacy of phenylpropanoids of plant origin against planktonic cells, important virulence factors, and biofilm forms of C. albicans. Standard susceptibility testing protocol was used to evaluate the activities of 13 phenylpropanoids against planktonic growth. Their effects on adhesion and yeast-to-hyphae morphogenesis were studied in microplate-based methodologies. An in vitro biofilm model analyzed the phenylpropanoid-mediated prevention of biofilm development and mature biofilms using XTT-metabolic assay, crystal violet assay, and light microscopy. Six molecules exhibited fungistatic activity at ${\leq}0.5mg/ml$, of which four were fungicidal at low concentrations. Seven phenylpropanoids inhibited yeast-to-hyphae transition at low concentrations (0.031-0.5 mg/ml), whereas adhesion to the solid substrate was prevented in the range of 0.5-2 mg/ml. Treatment with ${\leq}0.5mg/ml$ concentrations of at least six small molecules resulted in significant (p < 0.05) inhibition of biofilm formation by C. albicans. Mature biofilms that are highly resistant to antifungal drugs were susceptible to low concentrations of 4 of the 13 molecules. This study revealed phenylpropanoids of plant origin as promising candidates to devise preventive strategies against drug-resistant biofilms of C. albicans.

• Journal of Veterinary Science
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• v.19 no.6
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• pp.808-816
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• 2018

Bacterial biofilms have been demonstrated to be closely related to clinical infections and contribute to drug resistance. Berberine, which is the main component of Coptis chinensis, has been reported to have efficient antibacterial activity. This study aimed to investigate the potential effect of a combination of berberine with ciprofloxacin (CIP) to inhibit Salmonella biofilm formation and its effect on expressions of related genes (rpoE, luxS, and ompR). The fractional inhibitory concentration (FIC) index of the combination of berberine with CIP is 0.75 showing a synergistic antibacterial effect. The biofilm's adhesion rate and growth curve showed that the multi-resistant Salmonella strain had the potential to form a biofilm relative to that of strain CVCC528, and the antibiofilm effects were in a dose-dependent manner. Biofilm microstructures were rarely observed at $1/2{\times}MIC/FIC$ concentrations (MIC, minimal inhibition concentration), and the combination had a stronger antibiofilm effect than each of the antimicrobial agents used alone at $1/4{\times}FIC$ concentration. LuxS, rpoE, and ompR mRNA expressions were significantly repressed (p< 0.01) at $1/2{\times}MIC/FIC$ concentrations, and the berberine and CIP combination repressed mRNA expressions more strongly at the $1/4{\times}FIC$ concentration. The results indicate that the combination of berberine and CIP has a synergistic effect and is effective in inhibiting Salmonella biofilm formation via repression of luxS, rpoE, and ompR mRNA expressions.

• 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.28 no.2
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• pp.225-233
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• 2019

In this study, we investigated the in vitro anti-biofilm activities of plant extracts of chives (Allium tuberosum), garlic (Allium sativum), and radish (Raphanus sativus L.) against environment harmful bacteria (gram-positive Staphylococcus aureus and, gram-negative Salmonella typhimurium and Escherichia coli O157:H7). In the paper disc assay, garlic extracts exhibited the highest anti-biofilm activity. The Minimal Inhibitory Concentration (MIC) of all plant extracts was generally higher for gram-negative bacteria than it was for gram-positive bacteria. Gram-negative bacteria were more resistant to plant extracts. The tetrazolium dye (XTT) assay revealed that, each plant extract exhibited a different anti-biofilm activity at the MIC value depending on the pathogen involved. Among the plant extracts tested, garlic extracts (fresh juice and powder) effectively reduced the metabolic activity of the cells of food-poisoning bacteria in biofilms. These anti-biofilm activities were consistent with the results obtained through light microscopic observation. Though the garlic extract reduced biofilm formation for all pathogens tested, to elucidate whether this reduction was due to antimicrobial effects or anti-biofilm effects, we counted the colony forming units of pathogens in the presence of the garlic extract and a control antimicrobial drug. The garlic extract inhibited the E. coli O157:H7 biofilm effectively compared to the control antimicrobial drug ciprofloxacin; however, it did not inhibit S. aureus biofilm significantly compared to ciprofloxacin. In conclusion, garlic extracts could be used as natural food preservatives to prevent the growth of foodborne pathogens and elongater the shelf life of processed foods.

• Journal of Microbiology and Biotechnology
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• v.24 no.11
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• pp.1574-1582
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• 2014

Bacteria recognize changes in their population density by sensing the concentration of signal molecules, N-acyl-homoserine lactones (AHLs). AHL-mediated quorum sensing (QS) plays a key role in biofilm formation, so the interference of QS, referred to as quorum quenching (QQ), has received a great deal of attention. A QQ strategy can be applied to membrane bioreactors (MBRs) for advanced wastewater treatment to control biofouling. To isolate QQ bacteria that can inhibit biofilm formation, we isolated diverse AHL-degrading bacteria from a laboratory-scale MBR and sludge from real wastewater treatment plants. A total of 225 AHL-degrading bacteria were isolated from the sludge sample by enrichment culture. Afipia sp., Acinetobacter sp. and Streptococcus sp. strains produced the intracellular QQ enzyme, whereas Pseudomonas sp., Micrococcus sp. and Staphylococcus sp. produced the extracellular QQ enzyme. In case of Microbacterium sp. and Rhodococcus sp., AHL-degrading activities were detected in the whole-cell assay and Rhodococcus sp. showed AHL-degrading activity in cell-free lysate as well. There has been no report for AHL-degrading capability in the case of Streptococcus sp. and Afipia sp. strains. Finally, inhibition of biofilm formation by isolated QQ bacteria or enzymes was observed on glass slides and 96-well microtiter plates using crystal violet staining. QQ strains or enzymes not only inhibited initial biofilm development but also reduced established biofilms.

• Journal of Food Hygiene and Safety
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• v.37 no.4
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• pp.285-295
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• 2022

The ability of volatile components of essential oils (EO) from cinnamon, clove, and lemongrass to inhibit biofilms formed on polyethylene and stainless steel by six types of food poisoning bacteria was investigated. The main components of cinnamon EO were identified as cinnamaldehyde (38.30%), linalool (9.61%), β-caryophyllene (8.90%), and 1,3,4-eugenol (8.19%). 1,3,4-Eugenol (61.84%) was the dominant component of clove EO. The major component of lemongrass EO was citral. Citral is a natural mixture of two isomeric acyclic monoterpene aldehydes: geranial (trans-citral, 19.11%) and neral (cis-citral, 19.23%). Among these major compounds, cinnamaldehyde, linalool, eugenol, and citral exhibited comparatively strong antimicrobial activity in the disc diffusion assay. Treatments with 0.1% eugenol and citral were highly effective on biofilm inhibition on both tested surfaces. Cinnamaldehyde (0.1%) was effective against biofilm formation by Listeria monocytogenes ATCC 19112 and Staphylococcus aureus KCCM 11812. These results suggested the potential of cinnamaldehyde, eugenol, and citral treatments in inhibiting the formation of biofilms by food poisoning bacteria.