• Restorative Dentistry and Endodontics
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• 제44권1호
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• pp.4.1-4.10
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• 2019

Objectives: Biofilm formation is critical to dental caries initiation and development. The aim of this study was to investigate the effects of nicotine exposure on Streptococcus mutans (S. mutans) biofilm formation concomitantly with the inhibitory effects of sodium chloride (NaCl), potassium chloride (KCl) and potassium iodide (KI) salts. This study examined bacterial growth with varying concentrations of NaCl, KCl, and KI salts and nicotine levels consistent with primary levels of nicotine exposure. Materials and Methods: A preliminary screening experiment was performed to investigate the appropriate concentrations of NaCl, KCl, and KI to use with nicotine. With the data, a S. mutans biofilm growth assay was conducted using nicotine (0-32 mg/mL) in Tryptic Soy broth supplemented with 1% sucrose with and without 0.45 M of NaCl, 0.23 M of KCl, and 0.113 M of KI. The biofilm was stained with crystal violet dye and the absorbance measured to determine biofilm formation. Results: The presence of 0.45 M of NaCl, 0.23 M of KCl, and 0.113 M of KI significantly inhibited (p < 0.05) nicotine-induced S. mutans biofilm formation by 52%, 79.7%, and 64.1%, respectively. Conclusions: The results provide additional evidence regarding the biofilm-enhancing effects of nicotine and demonstrate the inhibitory influence of these salts in reducing the nicotine-induced biofilm formation. A short-term exposure to these salts may inhibit S. mutans biofilm formation.

• Biotechnology and Bioprocess Engineering:BBE
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• 제3권1호
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• pp.6-10
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• 1998

A plasmid vector containing two reporter genes, mer-lux and lac-GFP, was transformed to both Escherichia coli and Pseudomonas putida. Their cellular activities and biofilm characteristics were investigated in flow-cell units by measuring bioluminescent lights and fluorescent levels of GFP. Bioluminescence was effective to monitor temporal cell activities, whereas fluorescent level of GFP was useful to indicate the overall cell activities during biofilm development. The light production rates of E. coli and P. putida cultures were dependent upon concentrations of HgCl2. Mercury molecules entrapped in P. putida biofilms were hardly washed out in comparison with those in E. coli biofilms, indicating that P. putida biofilms may have higher affinity to mercury molecules than E. coli biofilms. It was observed that P. putida expressed GFP cDNA in biofilms but not in liquid cultures. This may indicate that the genetic mechanisms of P. putida were favorably altered in biofilm conditions to make a foreign gene expression possible.

• 한국환경보건학회지
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• 제37권2호
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• pp.124-132
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• 2011

Biofilm and aeration tank of pilot and full RABC (rotating activated Bacillus contactor) plant were analyzed to characterize and determine bacterial community structure in food wastewater treatment system at winter. Concentration of heterotrophic bacteria and Bacillus group was $10^7$ and $10^5$ CFU/ml, respectively, at biofilm of pilot-plant while others represented $10^6$ and $10^4$ CFU/ml, respectively. Five and eight phyla were detected at biofilm of pilot- and full-plant, respectively, by 16S rDNA sequencing. Biofilm of pilot-plant was dominated by ${\beta}$-Proteobacteria (38.8%), ${\gamma}$-Proteobacteria (22.4%), and Bacteroidetes (12.2%), and the most dominant genus was Zoogloeae genus (22.4%). Candidate division TM7 (12.5%) was only detected at biofilm of full-plant and it was dominated by Bacteroidetes (33.3%), ${\gamma}$-Proteobacteria (29.2%), and ${\beta}$-Proteobacteria (20.8%). Clostridium genus specific primer set enabled to detect the sequences of Clostridium genus. These suggested that anaerobic and aerobic bacteria were coexisted even from the initial period of biofilm formation and ${\beta}$-Proteobacteria, ${\gamma}$-Proteobacteria and Bacteroidetes were major phyla in biofilm of food wastewater treatment system at winter.

• Journal of Microbiology and Biotechnology
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• 제26권9호
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• pp.1593-1604
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• 2016

Recently, bacterial quorum quenching (QQ) has been proven to have potential as an innovative approach for biofouling control in membrane bioreactors (MBRs) for advanced wastewater treatment. Although information regarding the microbial community is crucial for the development of QQ strategies, little information exists on the microbial ecology in QQ-MBRs. In this study, the microbial communities of biofilm were investigated in relation to the effect of QQ on anoxic/oxic MBRs. Two laboratory-scale MBRs were operated with and without QQ-beads (QQ-bacteria entrapped in beads). The transmembrane pressure increase in the QQ-MBRs was delayed by approximately 100-110% compared with conventional- and vacant-MBRs (beads without QQ-bacteria) at 45 kPa. In terms of the microbial community, QQ gradually favored the development of a diverse and even community. QQ had an effect on both the bacterial composition and change rate of the bacterial composition. Proteobacteria and Bacteroidetes were the most dominant phyla in the biofilm, and the average relative composition of Proteobacteria was low in the QQ-MBR. Thiothrix sp. was the dominant bacterium in the biofilm. The relative composition of Thiothrix sp. was low in the QQ-MBR. These findings provide useful information that can inform the development of a new QQ strategy.

• 한국미생물·생명공학회지
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• 제50권4호
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• pp.548-556
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• 2022

Quorum quenching 활성을 나타내는 acylase 효소는 생물막 형성에 중요한 영향을 미친다. 본 연구에서는 acylase 처리 조건(acylase 처리 시기 및 기간)이 다종 생물막(multispecies biofilm) 형성에 미치는 영향을 규명하였다. 서로 다른 속(genus)에 속하는 9종의 박테리아로 구성된 컨소시엄을 사용하여 acylase 처리 조건에 따른 10가지 episode에서 다양한 acylase 농도(1, 5, 10, 20, 50 mg·l^-1)에 따라 5일 동안 생물막을 형성시켰다. 각 농도별로 5일 동안 acylase를 처리한 조건에서 acylase의 농도가 증가함에 따라 생물막 형성은 억제되었다(linear regression, Y = -0.05· x + 2.37, p < 0.05, R² = 0.88). 모든 acylase 농도 조건에서 acylase를 생물막이 형성된 후에 처리한 경우(L1-L4)에 비해서 생물막 형성 시작 단계에 처리한 경우(B1-B4) 생물막 형성이 더 효과적으로 억제되었다(p < 0.05). ANOVA 결과에 따르면 acylase 10 mg·l^-1 이상 농도에서 acylase 처리 기간(period)은 acylase 처리 시기(application timings, beginning or later)에 상관없이 생물막 형성에 영향을 미쳤다(p < 0.05). 각 농도별 처리 시기(L1-L4 또는 B1-B4)에서 처리 기간과 생물막 형성 사이의 선형 회귀 분석 결과에 따르면 acylase 10 mg·l^-1 이상 농도에서 acylase 처리 기간이 증가함에 따라 생물막 형성은 억제되었다(p < 0.05, 20 mg·l^-1 농도의 L1-L4 제외). 시간에 따른 생물막 형성 결과에 따르면 모든 10가지 episode에서 생물막은 시간에 따라 점차 증가했으며(p < 0.05), 배양 시간과 생물막 형성 사이의 선형 회귀 분석 기울기 값은 acylase를 생물막 형성 시작 단계에 처리했을 때 더 낮게 나타났다(p < 0.05). 본 연구 결과는 생물막 형성 억제에 대한 acylase의 처리 시기 및 기간의 중요성을 시사한다.

• Journal of Microbiology and Biotechnology
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• 제26권11호
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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 Dairy Science and Biotechnology
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• 제33권2호
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• pp.139-151
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• 2015

Biofilms are aggregates of microorganisms present in a self-produced matrix of extracellular polymeric substance (EPS) adhered to a surface. Formation of a biofilm in the environment on farms and in dairy plants comprises several stages: attachment, growth (development), and detachment. Generally, biofilms are harmful to humans and need to be controlled. Stainless steel (SS) surfaces that are untreated or are scratched comprise substrata that are especially vulnerable to biofilm formation; therefore, SS surfaces should be polished and sanitized. Various approaches are available for the destruction of biofilms; cleaning-in-place (CIP) is the method mainly used in dairy plants. Further study on optimum detergents, cleaning conditions, and methods for this purpose is needed.

• 한국미생물·생명공학회지
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• 제47권4호
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• pp.473-486
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• 2019

In the aquatic environment, microorganisms are predominantly organized as biofilms. Biofilms are formed by the aggregation of microbial cells and are surrounded by a matrix of extracellular polymeric substances (EPS) secreted by the microbial cells. Biofilms are attached to various surfaces, such as the living tissues, indwelling medical devices, and piping of the industrial potable water system. Biofilms formed from a single species has been extensively studied. However, there is an increased research focus on multispecies biofilms in recent years. It is important to assess the microbial mechanisms underlying the regulation of multispecies biofilm formation to determine the drinking water microbial composition. These mechanisms contribute to the predominance of the best-adapted species in an aquatic environment. This review focuses on the interactions in the multispecies biofilms, such as coaggregation, co-metabolism, cross-species protection, jamming of quorum sensing, lateral gene transfer, synergism, and antagonism. Further, this review explores the dynamics and the factors favoring biofilm formation and pathogen transmission within the drinking water distribution systems. The understanding of the physiology and biodiversity of microbial species in the biofilm may aid in the development of novel biofilm control and drinking water disinfection processes.

• 대한약침학회지
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• 제25권1호
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• pp.37-45
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• 2022

Objectives: The quorum-sensing-inhibitory and anti-biofilm activities of the methanol extract of E. globulus leaves were determined against clinically isolated multidrug-resistant Pseudomonas aeruginosa. Methods: The preliminary anti-quorum-sensing (AQS) activity of eucalyptus was investigated against a biosensor strain Chromobacterium violaceum ATCC 12472 (CV12472) by using the agar well diffusion method. The effect of sub-minimum inhibitory concentrations (sub-MICs) of the methanol extract of eucalyptus on different quorum-sensing-regulated virulence factors, such as swarming motility, pyocyanin pigment, exopolysaccharide (EPS), and biofilm formation, against clinical isolates (CIs 2, 3, and 4) and reference PA01 of Pseudomonas aeruginosa were determined using the swarm diameter (mm)-measurement method, chloroform extraction method, phenol (5%)-sulphuric acid (concentrated) method, and the microtiter plate assay respectively, and the inhibition (%) in formation were calculated. Results: The preliminary AQS activity (violacein pigment inhibition) of eucalyptus was confirmed against Chromobacterium violaceum ATCC 12472 (CV12472). The eucalyptus extract also showed concentration-dependent inhibition (%) of swarming motility, pyocyanin pigment, EPS, and biofilm formation in different CIs and PA01 of P. aeruginosa. Conclusion: Our results revealed the effectiveness of the E. globulus extract for the regulation of quorum-sensing-dependent virulence factors and biofilm formation at a reduced dose (sub-MICs) and suggest that E. globulus may be a therapeutic agent for curing and controlling bacterial infection and thereby reducing the possibility of resistance development in pathogenic strains.

• Journal of Microbiology and Biotechnology
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• 제24권9호
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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.