수처리용 분리막 시스템의 경우, 생물막 형성됨에 따라 분리막의 효율성이 저해된다. 이러한 문제를 완화 시키고자 생물막 형성을 방해하는 정족수 감지 억제 기술이 연구 되고 있다. 본 연구에서는 미생물의 내생기작과 외생기작을 각각 적용하여 생물막 저감 효과를 한 줄로 구성된 중공사막 모듈과 다발형 모듈에서 비교하였다. 그 결과 한 줄로 구성된 모듈에서는 내생기작과 외생기작이 미생물 생성 억제 성능에 큰 차이가 없었지만, 다발형 모듈에서는 외생 기작이 더 효과적인 것으로 나타났다. 따라서 다발형 모듈로 구성된 실제 중공사 수처 리 시스템에서는 외생 기작을 가진 정족수 감지 억제 미생물을 사용하여 분리 막 표면에 생기는 생물막을 저감하는 것이 내생 기작을 가진 미생물을 적용하는 것 보다 효율적인 것으로 보여진다.

Due to ever increasing trends in food safety, food manufacturers should take sanitary/ hygienic processing/packaging into key consideration. Minimizing the attachment of spoilage and pathogenic organisms to food contact surfaces and packaging materials is one of the major challenges in the fields of food science and biosafety. The bacteria adhering to the surface can produce subsequent hard-to-remove biofilms that potentially cause cross-contamination of processed foods. The contamination of spoilage to the food leads to shorten product shelf-life and, if foodborne pathogens are present, the consumption of contaminated food may possess a health hazard to consumers. In particular, a variety of surfaces in food packaging are vulnerable to development of biofilms as bacterial community is formed by adhering pathogens from native microflora in raw materialsfor prolonged contact time. It is now recognized that 80% of food outbreaks are related to biofilms. Therefore, inhibition of initial adhesion of biofilm-forming pathogenic cells on food contact surface is critically needed to minimize foodborne pathogen outbreaks.Bacterial adhesion is highly influenced by the substratum topography, i.e. roughness and porosity. Recent advancements in fabrication have made it possible to create well-organized nanofeatures (i.e. nanoporous and nanopillared) uniformly over a large surface area of a metal specimen. Nanosmooth (control) and nanoporous stainless steel foil surfaces were fabricated by precisely anodizing the degreased specimen in a 5% vol. of perchloric acid in anhydrous ethylene glycol. The applied voltage and anodization time were varied to obtain different pore diameters. The presences of 50 and 80 nm nanoporous patterns significantly inhibited the adhesion of L. monocytogenes by 2.0 to 2.3 log-cycles, depending on the pore diameters. It was found that nanoscale surface patterning and treatments are capable of enabling precise controls of molecular, physical, and biochemical interactions that govern bacterial adhesion to the solid substratum.

Cronobactersakazakii is a newly emerging high hazard pathogen, which causes encephalomeningitis and necrotic colitis. Recently, successful biocontrol of harmful microorganisms in several foods through the use of bacteriophages has been reported. In this study, bacteriophages were isolated from kimchi and sewages. Morphological analysis by TEM indicated that phages belonged to the Myoviridae family. In case of heat stability, KCES2 and ESP 2949-2 phages were susceptible to temperatures above 70oC. KCES2 and ESP 2949-2 phages inhibited the growth of C. sakazakii in culture broth. When KCES2 and ESP 2949-2 phages were applied to biofilm-formed C. sakazakii, C. sakazakii was efficiently reduced. Therefore, newly isolated KCES2 and ESP 2949-2 phage for C. sakazakii might effectively reduce C. sakazakii in various foods.

Minimal inhibitory concentration (MIC) is the lowest antibiotic concentration that inhibits the visible growth of bacteria. Sub-minimal inhibitory concentration (Sub-MIC) is defined as the concentration of an antimicrobial agent that does not have an effect on bacterial growth but can alter bacterial biochemistry, thus reducing bacterial virulence. Many studies have confirmed that sub-MICs of antibiotics can inhibit bacterial virulence factors. However, most studies were focused on Gram-negative bacteria, while few studies on the effect of sub-MICs of antibiotics on Gram-positive bacteria. In this study, we examined the influence of sub-MICs of doxycycline, tetracycline, penicillin and amoxicillin on biofilm formation and coaggregation of Streptococcus gordonii, Streptococcus mutans, Actinomyces naeslundii, and Actinomyces odontolyticus. In this study, incubation with sub-MIC of antibiotics had no effect on the biofilm formation of S. gordonii and A. naeslundii. However, S. mutans showed increased biofilm formation after incubation with sub-MIC amoxicillin and penicillin. Also, the biofilm formation of A. odontolyticus was increased after incubating with sub-MIC penicillin. Coaggregation of A. naeslundii with S. gordonii and A. odontolyticus was diminished by sub-MIC amoxicillin. These observations indicated that sub-MICs of antibiotics could affect variable virulence properties such as biofilm formation and coaggregation in Gram-positive oral bacteria.

Biofilms of oral microbes can cause various diseases in the oral cavity, such as dental caries, periodontitis and mucosal disease. Electrolyzed water generated by an electric current passed via water using a metal electrode has an antimicrobial effect on pathogenic bacteria which cause food poisoning. This study investigated the antimicrobial activity of electrolyzed waters using various metal electrodes on the floatage and biofilms of oral microbes. The electrolyzed water was generated by passing electric current using copper, silver and platinum electrodes. The electrolyzed water has a neutral pH. Streptococcus mutans, Porphyromonas gingivalis and Tannerella forsythia were cultured, and were used to form a biofilm using specific media. The floatage and biofilm of the microbes were then treated with the electrolyzed water. The electrolyzed water using platinum electrode (EWP) exhibited strong antimicrobial activity against the floatage and biofilm of the oral microbes. However, the electrolyzed water using copper and silver electrodes had no effect. The EWP disrupted the biofilm of oral microbes, except the S. mutans biofilm. Comparing the different electrolyzed waters that we created the platinum electrode generated water may be an ideal candidate for prevention of dental caries and periodontitis.

The purpose of this study was to assess the efficacy of photodynamic therapy (PDT) using erythrosine and a halogen light source to treat a biofilm formed on a machined surface titanium disk in vivo. Ten volunteers carried an acrylic appliance containing six machined surface titanium disks on the upper jaw over a period of five days. After the five days of biofilm formation period, the disks were removed. PDT using 20 μM erythrosine and halogen light was then applied to the biofilms formed on the disks. Experimental samples were divided into a negative control group (no erythrosine and no irradiation), E0 group (erythrosine 60s + no irradiation), E30 group (erythrosine 60s + halogen light 30s), and E60 group (erythrosine 60s + halogen light 60s). Following PDT, the bacteria in the biofilm were found to be detached from each disk. Each suspension with detached bacteria were diluted and cultivated on a blood-agar plate for five days under anaerobic conditions. The cultivated bacterial counts in the E60 group were significantly lower than the control group (86.4%) or E0 group (76.7%). In the experimental groups also, the light exposure time and bacterial counts showed a negative correlation. In conclusion, PDT using erythrosine and halogen light has bactericidal effects on biofilms formed on a titanium disk in vivo. Notably, applying 20 μM erythrosine and 60 seconds of halogen light irradiation had a significantly potent effect.

Bacillus cereus is widely distributed on various foods and is known to cause clinical infections, food poisoning toxin induced diarrhea and vomiting. In this study, B. cereus group detected and analyzed rice, rice bran, and biofilm characterization of B. cereus confirmed. B. cereus was identified in approximately 34.6% of brown rice and 50.0% of rice bran. B. thuringiensis was detected in 3.9% of brown rice and 23% of rice bran, and B. mycoides was isolated from rice bran. The microtiter plate assay detected differences in biofilm-forming ability among B. cereus group isolates. Biofilm of B. cereus seemed to increase the MIC values of antimicrobial agent and antibiotic compounds compared with planktonic cells. Therefore, sufficient attention should be given to good manufacturing practice and good agriculture practice to avoid contamination of B. cereus group raw material including rice.

The present study was conducted to investigate serotype distribution and biofilm formation of Actinobacillus (A.) pleuropneumoniae isolated from pneumonic lungs of pigs. A total of 37 A. pleuropneumoniae were isolated between January 2009 and June 2010. Serotypes of A. pleuropneumoniae isolates were determined using two different PCRs. The majority of isolates belonged to serotype 5 (n=31, 83.8%), and the others belonged to serotype 1 (n=4, 10.8%), and 2 (n=2, 5.4%), respectively. The ability of biofilm formation of the isolates was also determined by quantitative microtiter plate assay. Biofilm formation was observed in both 23 (62.2%) of the 37 field isolates and seven (43.8%) of the 16 reference strains. On the other hand, biofilm formation was various according to the serotypes: 20 (64.5%) of serotype 5, and three (75.0%) of serotype 1. However, two isolates of serotype 2 did not produce the biofilm in this study. Consequently, A. pleuropneumoniae serotype 5 was the most frequently detected (83.8% of the isolates), and 23 (62.2%) of 37 isolates exhibited biofilm positive phenotype in this study.

A number of bacterial species coexist in oral cavities as a biofilm rather than a planktonic arrangement. By forming an oral biofilm with quorum sensing properties, microorganisms can develop a higher pathogenic potential and stronger resistance to the host immune system and antibiotics. Hence, the inhibition of biofilm formation has become a major research issue for the future prevention and treatment of oral diseases. In this study, we investigated the effects of pentose on biofilm formation and phenotypic changes using wild type oral bacteria obtained from healthy human saliva. D-ribose and D-arabinose were found to inhibit biofilm formation, but have no effects on the growth of each oral bacterium tested. Pentoses may thus be good candidate biofilm inhibitors without growth-inhibition activity and be employed for the future prevention or treatment of oral diseases.

In our present study, we investigated the effects of continentalic acid on Streptococcus mutans (S. mutans) biofilm. Methanol extract of Aralia continentalis (A. continentalis) was suspended in water and sequentially partitioned with CHCl3, ethyl acetate (EtOAc), and n-butanol (n-BuOH). The CHCl3 fraction showed the highest activity and an antibacterial compound against S. mutans was isolated from this preparation through various chromatography methods by bioassay guided fractionation. MS, 1H - NMR and 13C-NMR analysis showed that the active principle was continentalic acid which was confirmed to show significant inhibitory effects against S. mutans biofilm. These results may provide some scientific rationale for the traditional use these extracts for the treatment of dental diseases.

Most oral microorganisms exist as biofilms which initiate formation via the attachment of an early colonizer to host proteins on the tooth surface. Fusobacterium nucleatum act as a bridge between early and late colonizers. Dental biofilms eventually comprise dental pathogens such as Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia. To evaluate the effects of mutual interactions between oral bacteria on the growth of biofilms, periodontopathogens were co-cultured with a 0.4 μm barrier. Streptococcus gordonii inhibited the growth of F. nucleatum and periodontopathogens. However, F. nucleatum, P. gingivalis and T. denticola activated the growth of other bacteria. A co-culture system of early and late colonizers could be a useful tool to further understand bacterial interactions during the development of dental biofilm.

Denitrification activity of microbial communities in sand-gravel sediments and biofilms developing on a concrete riverbed of an urban small stream, Nogawa River, was measured monthly by laboratory incubations after the acetylene inhibition technique during une year from December 1987. The annual mean denitrification rate of the biofilm samples, 1.5 mgN m-2 h-1. was higtrer than the rate ut the sandgravel sediment samples. 0.8 mgN m-2 h-1. Contrary, ratios of the denitrified nitrogen to the total inorganic nitrogen fluxes was lower at the stretch where an attached microbial community developed un the concrete riverbed the stretch with sand-gravel riverbed. The longer residence times of water masses ut the site with a sand-gravel riverbed is responsible for the above relationship. Based on the average denitrifieation rates, if all the channel of Nogawa River were covered with concrete, the proportion of denitrified nitrogen to the input nitrogen should decrease to 1.3 %, whereas the ratio is 2.5% when the riverbed is composed of sands and gravel. The results suggest that the efficiency of nitrogen removal by benthic denitrification is strongly affected by the structure of a channel of the river.

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.

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.

Haloacetic acids (HAAs) concentrations have been observed to decreased at drinking water distribution system extremities. This decrease is associated with microbiological degradation by pipe wall biofilm. The objective of this study was to evaluate HAAs degradation in a drinking water system in the presence of a biofilm and to identify the factors that influence this degradation. Degradation of monochloroacetic acid (MCAA), dichloroacetic acid (DCAA) and trichloroacetic acid (TCAA) was observed in a simulated distribution system. The results obtained showed that different parameters came into play simultaneously in the degradation of HAAs, including retention time, water temperature, biomass, and composition of organic matter. Seasonal variations had a major effect on HAAs degradation and biomass quantity (ATP concentration) was lower by 25% in the winter compared with the summer.

In this study we followed biofilm formation and development in a granular activated carbon (GAC) filter on pilot-scale during the 12 months of operation. GAC particles and water samples were sampled from four different depths (-5, -25, -50 and –90 cm from surface of GAC bed) and attached biomass were measured with adenosine tri-phosphate (ATP) analysis and heterotrophic plate count (HPC) method. The attached biomass accumulated rapidly on the GAC particles of top layer throughout all levels in the filter during the 160 days (BV 23,000) of operation and maintained a steady-state afterward. During steady-state, biomass (ATP and HPC) concentrations of top layer in the BAC filer were 2.1 μg·ATP/g·GAC and 3.3×108 cells/g·GAC, and 85%, 83% and 99% of the influent total biodegradable dissolved organic carbon (BDOCtotal), BDOCslow and BDOCrapid were removed, respectively. During steady-state process, biomass (ATP and HPC) concentrations of middle layer (-50 cm) and bottom layer (-90 cm) in the BAC filter were increased consistently. Biofilm development (growth rate) proceed highest rate in the top layer of filter (μATP = 0.73 day-1; μHPC = 1,74 day-1) and 78%∼87% slower in the bottom layer (μATP = 0.14 day-1; μHPC = 0.34 day-1). This study shows that the combination of different analytical methods allows detailed quantification of the microbiological activity in drinking water biofilter.

A fixed biofilm reactor system composed of anaerobic, anoxic(1), anoxic(2), aerobic(1) and aerobic(2) reactor was packed with synthetic activated ceramic (SAC) media and adopted to reduce the inhibition effect of low temperature on nitrification activities. The changes of nitrification activity at different wastewater temperature were investigated through the evaluation of temperature coefficient, volatile attached solid (VAS), specific nitrification rate and alkalinity consumption. Operating temperature was varied from 20 to 5 ℃. In this biofilm system, the specific nitrification rates of 15 ℃, 10 ℃ and 5 ℃ were 0.972, 0.859 and 0.613 when the specific nitrification rate of 20 ℃ was assumed to 1.00. Moreover the nitrification activity was also observed at 5 ℃ which is lower temperature than the critical temperature condition for the microorganism of activated sludge system. The specific amount of volatile attached solid (VAS) on media was maintained the range of 13.6-12.5 mg VAS/g media at 20~10 ℃. As the temperature was downed to 5 ℃, VAS was rapidly decreased to 10.9 mg VAS/g media and effluent suspended solids was increased from 3.2 mg/L to 12.0 mg/L due to the detachment of microorganism from SAC media. And alkalinity consumption was lower than theoretical value with 5.23 mg as CaCO3/mg NH4 +-N removal at 20 ℃. Temperature coefficient (θ) of nitrification rate (20 ℃ ~ 5 ℃) was 1.033. Therefore, this fixed film nitrogen removal process showed superior stability for low temperature condition than conventional suspended growth process.

The vertical distributions of nitrifying bacteria in aerobic fixed biofilm were investigated to evaluate the relationship between nitrification performance and microbial community at different HRT. Fluorescent in situ hybridization (FISH) and portable ion selective microelectrode system were adopted to analyze microbial communities and ions profiles according to the biofilm depth. Cilia media packed MLE (Modified Ludzack-Ettinger) like reactor composed of anoxic, aerobic Ⅰ/Ⅱ was operated with synthetic wastewater having COD 200 mg/L and NH4 +-N 40 mg/L at HRT of 6 hrs and 4 hrs. Total biofilm thickness of aerobic Ⅰ, Ⅱ reactor at 4 hrs condition was over two times than that of 6 hrs condition due to the sufficient substrate supply at 4 hrs condition (6 hrs; aerobic Ⅰ 380 ㎛ and Ⅱ 400 ㎛, 4 hrs; aerobic Ⅰ 830 ㎛ and Ⅱ 1040 ㎛). As deepen the biofilm detection point, the ratio of ammonia oxidizing bacteria (AOB) was decreased while the ratio of nitrite oxidizing bacteria (NOB) was maintained similar distribution at both HRT condition. The ratio of AOB was higher at 4 hrs than 6 hrs condition and NH4 +-N removal efficiency was also higher at 4 hrs with 89.2% than 65.4% of 6 hrs. However, the ratio of NOB was decreased when HRT was reduced from 6 hrs to 4 hrs and NO2 --N accumulation was observed at 4 hrs condition. Therefore, it is considered that insufficient HRT condition could supply sufficient substrate and enrichment of AOB in all depth of fixed biofilm but cause decrease of NOB and nitrite accumulation.