• Journal of Periodontal and Implant Science
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• v.44 no.2
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• pp.79-84
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• 2014

Purpose: While single-species biofilms have been studied extensively, we know notably little regarding multispecies biofilms and their interactions. The purpose of this study was to develop and evaluate an in vitro multispecies dental biofilm model that aimed to mimic the environment of chronic periodontitis. Methods: Streptococcus gordonii KN1, Fusobacterium nucleatum ATCC23726, Aggregatibacter actinomycetemcomitans ATCC33384, and Porphyromonas gingivalis ATCC33277 were used for this experiment. The biofilms were grown on 12-well plates with a round glass slip (12 mm in diameter) with a supply of fresh medium. Four different single-species biofilms and multispecies biofilms with the four bacterial strains listed above were prepared. The biofilms were examined with a confocal laser scanning microscope (CLSM) and scanning electron microscopy (SEM). The minimum inhibitory concentrations (MIC) for four different planktonic single-species and multispecies bacteria were determined. The MICs of doxycycline and chlorhexidine for four different single-species biofilms and a multispecies biofilm were also determined. Results: The CLSM and SEM examination revealed that the growth pattern of the multispecies biofilm was similar to those of single-species biofilms. However, the multispecies biofilm became thicker than the single-species biofilms, and networks between bacteria were formed. The MICs of doxycycline and chlorhexidine were higher in the biofilm state than in the planktonic bacteria. The MIC of doxycycline for the multispecies biofilm was higher than were those for the single-species biofilms of P. gingivalis, F. nucleatum, or A. actinomycetemcomitans. The MIC of chlorhexidine for the multispecies biofilm was higher than were those for the single-species biofilms of P. gingivalis or F. nucleatum. Conclusions: To mimic the natural dental biofilm, a multispecies biofilm composed of four bacterial species was grown. The 24-hour multispecies biofilm may be useful as a laboratory dental biofilm model system.

• Food Science of Animal Resources
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• v.29 no.6
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• pp.702-708
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• 2009

We investigated biofilm formation in various media, growth in low pH, and the hemolytic activity of 14 strains of Cronobacter spp. (Enterobacter sakazakii) isolated from a variety of foods including powdered infant formula (n=75), infant cereal (n=100), honey (n=30), and other infant foods (n=100) in Korea. The Cronobacter spp. adhered and formed biofilms on polyethylene, and a greater extent of biofilm was observed in nutrient-rich media. No clear difference in biofilm-forming ability was noted among the media constituents and the pattern of biofilm formation was strain-dependent. Seven strains out of 14 strains (50%) grew at pH 4.1, indicating that the acid resistance of these Cronobacter spp. isolated in Korea was relatively low. Hemolytic activity was not observed in any of the strains. This study provides basic information for the physiological and biochemical characteristics of Cronobacter spp. isolated from a variety of infant foods in Korea.

• International Journal of Oral Biology
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• v.43 no.4
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• pp.171-175
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• 2018

Quorum sensing (QS) is a cell density-dependent communication mechanism between bacteria through small signaling molecules. When the number of QS signaling molecules reaches a threshold, they are transported back into the cells or recognized by membrane-bound receptors, triggering gene expression which affects various phenotypes including bioluminescence, virulence, adhesion, and biofilm formation. These phenotypes are beneficial for bacterial survival in harsh environments. This review summarizes the application of QS inhibitors for control of biofilm formation and virulence expression of periodontal pathogens.

• Journal of Food Hygiene and Safety
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• v.36 no.5
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• pp.430-439
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• 2021

Essential oils with excellent antibacterial activity were used to study the inhibitory effect against the six types of food poisoning biofilms formed on the surfaces of polyethylene (PE) and stainless steel (SS) that are widely used for food processing instruments and containers. The antibacterial activity of 20 kinds of essential oils was tested using the disk diffusion method. The result showed the degree of antibacterial activity in the following order: cinnamon> clove> lemongrass> peppermint> pine needle (highest to lowest). The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of cinnamon and clove oil were in the range of 0.63-1.25 mg/mL and 1.25-2.50 mg/mL, respectively. The MIC and MBC of lemongrass oil were 1.25-2.50 mg/mL and 2.50-5.00 mg/mL, respectively, showing slightly less antibacterial activity. Although the preventive effect of three types of essential oils on the biofilm formation differed slightly depending on food poisoning bacteria, PE, and SS, it was found that the precoating of 0.5% cinnamon, clove, and lemongrass oil on the PE and SS affects the formation of biofilm. Increased essential oil concentration significantly inhibited the biofilm formation for all food poisoning bacteria (P<0.05), and biofilms of Listeria monocytogenes and Staphylococcus aureus were not formed when treated with 0.5% cinnamon and clove oil. The elimination effect of food poisoning bacteria biofilms formed on the surfaces of PE and SS differed depending on the type of food poisoning bacteria. Still, the biofilm elimination effect increased as the essential oil concentration increased, and the biofilm elimination rate of clove oil was generally high. Therefore, this study found that the cinnamon and clove essential oils (0.5%) are suitable natural materials that effectively prevent, inhibit, and remove the biofilms formed by the food poisoning bacteria on the surfaces of polyethylene and stainless steel.

• The Journal of Korean Academy of Prosthodontics
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• v.51 no.4
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• pp.245-251
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• 2013

Purpose: This study was conducted to compare in vivo biofilm formation on titanium surface and zirconia surface. Materials and methods: For biofilm formation on titanium and zirconia in oral cavity, after producing oral appliances using acrylic resin and orthodontic wire tailored to 9 subjects, we made titanium and zirconia specimens ($6mm{\times}6mm{\times}2mm$), fixed them on oral appliances and maintained them in oral cavity of test subjects for 24 and 72 hours. Test subjects who have equipped two pairs of specimens maintained oral hygiene not by using toothpaste but only by tooth brushing. After 24 and 72 hours, we removed and observed specimens through scanning electron microscopy (SEM). Results: Biofilm formation showed large deviation depending on individuals. For formation comparison between titanium and zirconia for 24 hours, zirconia showed less biofilm formation than titanium. Biofilm formation showed large deviation depending on individuals. As for formation comparison between zirconia and titanium, the degree of biofilm formation in zirconia was less than it was in titanium after a lapse of 24 hours. The result of biofilm formation in 72 hours trial show that zirconia has an inclination to formate less biofilm than it was in titanium. Conclusion: Based on the above results, we can conclude that early biofilm formation in oral cavity was influenced by difference of abutment materials.

• Journal of Microbiology and Biotechnology
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• v.18 no.8
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• pp.1459-1469
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• 2008

Using a rotating biological contactor modified with a sequencing bath reactor system (SBRBC) designed and operated to remove phosphate and nitrogen [58], the microbial community structure of the biofilm from the SBRBC system was characterized based on the extracellular polymeric substance (EPS) constituents, electron microscopy, and molecular techniques. Protein and carbohydrate were identified as the major EPS constituents at three different biofilm thicknesses, where the amount of EPS and bacterial cell number were highest in the initial thickness of 0-100${\mu}m$. However, the percent of carbohydrate in the total amount of EPS decreased by about 11.23%, whereas the percent of protein increased by about 11.15% as the biofilm grew. Thus, an abundant quantity of EPS and cell mass, as well as a specific quality of EPS were apparently needed to attach to the substratum in the first step of the biofilm growth. A FISH analysis revealed that the dominant phylogenetic group was $\beta$- and $\gamma$-Proteobacteria, where a significant subclass of Proteobacteria for removing phosphate and/or nitrate was found within a biofilm thickness of 0-250${\mu}m$. In addition, 16S rDNA clone libraries revealed that Klebsiella sp. and Citrobacter sp. were most dominant within the initial biofilm thickness of 0-250${\mu}m$, whereas sulfur-oxidizing bacteria, such as Beggiatoa sp. and Thiothrix sp., were detected in a biofilm thickness over 250${\mu}m$. The results of the bacterial community structure analysis using molecular techniques agreed with the results of the morphological structure based on scanning electron microscopy. Therefore, the overall results indicated that coliform bacteria participated in the nitrate and phosphorus removal when using the SBRBC system. Moreover, the structure of the biofilm was also found to be related to the EPS constituents, as well as the nitrogen and phosphate removal efficiency. Consequently, since this is the first identification of the bacterial community and structure of the biofilm from an RBC simultaneously removing nitrogen and phosphate from domestic wastewater, and it is hoped that the present results may provide a foundation for understanding nitrate and phosphate removal by an RBC system.

• 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 Korean Society of Environmental Engineers
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• v.28 no.1
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• pp.26-33
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• 2006

This study investigated the characteristics of the biofilm removal by free chlorine or monochloramine. The simulated drinking water distribution pipes on which biofilms had been formed were supplied with tap water containing 0.5, 1.0, 2.0 mg/L of free chlorine or monochloramine residuals. The biofilm removal was characterized by measurement of attached HPC and biomass on pipe surfaces. Chlorine was more effective in both inactivation of attached viable heterotrophic bacteria and removal of biofilm biomass compared to monochloramine. Biofilm matrix was not much eliminated from the surfaces by monochloramine disinfection. Free chlorine residual of 2.0 mg/L was found to be effective in biomass removal. However, biofilm level as low as $10CFU/cm^2$ of attached HPC and $5{\mu}g/cm^2$ of biomass still remained on the surfaces at 2.0 mg/L of chlorine residual. The measurement of biomass appeared to be a useful means in evaluating the characteristics of biofilm removal.

• Environmental Engineering Research
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• v.15 no.2
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• pp.71-78
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• 2010

To understand the effects of acclimation schemes on the formation of anode biofilms, different electrical performances are characterized in this study, with the roles of suspended and attached bacteria in single-chamber microbial fuel cells (MFCs). The results show that the generation of current in single-chamber MFCs is significantly affected by the development of a biofilm matrix on the anode surface containing abundant immobilized microorganisms. The long-term operation with suspended microorganisms was demonstrated to form a dense biofilm matrix that was able to reduce the activation loss in MFCs. Also, a Pt-coated anode was not favorable for the initial or long-term bacterial attachment due to its high hydrophobicity (contact angle = $124^{\circ}$), which promotes easy detachment of the biofilm from the anode surface. Maximum power ($655.0\;mW/m^2$) was obtained at a current density of $3,358.8\;mA/m^2$ in the MFCs with longer acclimation periods. It was found that a dense biofilm was able to enhance the charge transfer rates due to the complex development of a biofilm matrix anchoring the electrochemically active microorganisms together on the anode surface. Among the major components of the extracellular polymeric substance, carbohydrates ($85.7\;mg/m^2_{anode}$) and proteins ($81.0\;mg/m^2_{anode}$) in the dense anode biofilm accounted for 17 and 19%, respectively, which are greater than those in the sparse anode biofilm.

• Food Science of Animal Resources
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• v.34 no.2
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• pp.257-261
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• 2014

The objective of this study was to evaluate the effects of NaCl on the biofilm formations of the isolate from Staphylococcus aureus outbreaks linked to ham. The S. aureus ATCC13565 isolated from ham was exposed to NaCl concentrations of 0%, 2%, 4%, and 6% supplemented in tryptic soy broth (TSB) for 24 h at $35^{\circ}C$, followed by plating 0.1 mL of the culture on tryptic soy agar containing 0%, 2%, 4%, and 6% NaCl, respectively. After incubating at $35^{\circ}C$ for 24 h, the colonies on the plates were collected and diluted to $OD_{600}$ = 0.1. The diluents of S. aureus were incubated on a 96-well flat bottom plate containing TSB plus the appropriate NaCl concentrations, and the biofilm formation was quantified by crystal violet staining after being incubated at $35^{\circ}C$ for 9 h. Confocal laser scanning microscope (CLSM) was also used for visualizing the biofilm formation of S. aureus at NaCl concentrations of 0%, 2%, 4%, and 6%. The transcriptional analysis of biofilm-related genes, such as icaA, atl, clfA, fnbA, sarA, and rbf, was conducted by quantitative real-time PCR. Crystal violet staining and CLSM showed that the biofilm formations of S. aureus increased (p<0.05) along with the NaCl concentrations. Moreover, the expression of the icaA genes was higher at the NaCl concentrations of 4% and 6% as compared with 0% of NaCl by approximately 9-folds and 20-folds, respectively. These results indicated that the NaCl formulated in processed food may increase the biofilm formations of S. aureus by increasing the icaA gene expressions.