• Journal of Periodontal and Implant Science
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• 제44권2호
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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.

• 한국미생물·생명공학회지
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• 제23권4호
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• pp.492-498
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• 1995

Net type polyvinylidene chloride (PVDC) media and cillium type polyethlene polypropylene (PEPP) media were installed in the aereted submerged biofilm reactors. Synthetic phenol wastewater for feed were made to contain 1,480 mg of phenol per liter of water. The organic loading range of reactors were 0.439-0.456 kg COD/m$_{3}$, 0.882 - 0.919 kg COD/m$_{3}$ and 1.199-1.339 kg COD/m$_{3}$. Comparing PVDC to PEPP media, the bacterial number found in biofilm on PEPP were slightly higher. With the low temperature (10$\circ$C), the number of bacteria was some what deceered. Number of bacterial strains identified from PVDC were 23 and those from PEPP were 42. Genera identified in the PVDC media were Flavobacterium (47.8%), Unidentified (17.6%), Pseudomonas (13.0%), Micrococcus (8.7%) and Beggratoa (8.7%). Genera identified in the PEPP media reactor were Pseudomonas (35.7%), Alcaligenes (19.0%), Aeromonas (14.33%) and Micrococcus (11.9%), In the steady, state, a filamentous bacteria, type 1701 was identified in all of the reactors. Paramecium sp. and fungi were present in the PVDC media reactor. While, Difflugia sp, Paramecium sp. and fungi were found in the PEPP media reactor. The low diversity of protozoa was ascribed to high concentration of phenol.

• International Journal of Oral Biology
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• 제43권4호
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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.

• 한국축산식품학회지
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• 제40권4호
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• pp.668-674
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• 2020

Manuka honey (MH) has been shown anti-bacterial activity against several pathogenic bacteria. However, the inhibitory effect of MH on biofilm formation by Escherichia coli O157:H7 has not yet been examined. In this study, MH significantly reduced E. coli O157:H7 biofilm. Moreover, pre- and post-treatment with MH also significantly reduced E. coli O157:H7 biofilm. Cellular metabolic activities exhibited that the viability of E. coli O157:H7 biofilm cells was reduced in the presence of MH. Further, colony forming unit of MH-treated E. coli O157:H7 biofilm was significantly reduced by over 70%. Collectively, this study suggests the potential of anti-biofilm properties of MH which could be applied to control E. coli O157:H7.

• Journal of Microbiology and Biotechnology
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• 제15권6호
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• pp.1221-1228
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• 2005

Carbon electrode was applied to a wastewater treatment system as biofilm media. The spatial distribution of heterotrophic bacteria in aerobic wastewater biofilm grown on carbon electrode was investigated by scanning electron microscopy, atomic force microscopy, and biomass measurement. Five volts of electric oxidation and reduction potential were charged to the carbon anode and cathode of the bioelectrochemical system, respectively, but were not charged to electrodes of a conventional system. To correlate the biofilm architecture of bacterial populations with their activity, the bacterial treatment efficiency of organic carbons was measured in the bioelectrochemical system and compared with that in the conventional system. In the SEM image, the biofilm on the anodic medium of the bioelectrochemical system looked intact and active; however, that on the carbon medium of the conventional system appeared to be shrinking or damaging. In the AFM image, the thickness of biofilm formed on the carbon medium was about two times of those on the anodic medium. The bacterial treatment efficiency of organic carbons in the bioelectrochemical system was about 1.5 times higher than that in the conventional system. Some denitrifying bacteria can metabolically oxidize $H_{2}$, coupled to reduction of $NO_{3}^{-}\;to\;N_{2}$. $H_{2}$ was produced from the cathode in the bioelectrochemical system by electrolysis of water but was not so in the conventional system. The denitrification efficiency was less than $22\%$ in the conventional system and more than $77\%$ in the bioelectrochemical system. From these results, we found that the electrochemical coupling reactions between aerobic and anaerobic reactors may be a useful tool for improvement of wastewater treatment and denitrification efficiency, without special manipulations such as bacterial growth condition control, C/N ratio (the ratio of carbon to nitrogen) control, MLSS returning, or biofilm refreshing.

• 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.

• 한국환경과학회지
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• 제30권5호
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• pp.369-378
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• 2021

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.

• 한국미생물·생명공학회지
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• 제40권2호
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• pp.83-91
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• 2012

본 총설은 N-acyl-homoserine lactone (AHL)에 기반한 quorum sensing(QS)을 비롯한 다양한 QS 시스템 및 생물막 형성과의 관련성에 대한 연구 동향을 정리하였다. 또한 anti-QS으로서 quorum quenching 전략을 이용한 생물막 억제 연구 동향에 대해 중점적으로 서술하였다. 세균의 독특한 신호전달 체계인 QS는 AHL과 같은 특정한 신호분자의 농도에 의해 세균의 집단적 행동 양식이 결정되는 세포밀도-의존성 유전자 발현 조절 메커니즘이다. QS 시스템은 미생물의 부착 및 생물막 형성에 있어 중요한 역할을 한다. AI-1이나 AI-2에 의한 QS는 생물막 형성 과정에 필요한 세포외 다당류, 단백질, 세포 외 DNA 등 주요한 구성 성분 등의 생산뿐만 아니라, 세균의 운동성 조절, 부착, 생물막 해체 과정까지도 조절하는 기능을 한다. 일부 세균의 경우 QS시스템 이외에도 second messenger로 알려진 c-di-GMP에 의한 signaling이 QS와 서로 연결되어 생물막 형성이나 병독성과 같은 타깃들을 함께 조절한다. 생물막은 병원성 세균에 의한 감염 시 여러 가지 병독성 가운데 가장 중요한 요소 중 하나이기 때문에, 생물막 형성을 조절하는 QS를 차단하기 위한 다양한 anti-quorum sensing 전략이 연구되고 있다. Anti-QS 접근 방식은 의학적 이용뿐만 아니라 물에 노출되어있는 MBR을 비롯한 많은 산업적 장치 등에서 생물막 형성으로 인한 손상 및 오염을 방지하기 위해 쓰일 수 있다. Anti-QS 전략 중 신호분자인 AHL을 무력화 시키는 quorum quenching 효소(AHL-lactonase, AHL-acylase, oxidoreductas)를 이용하여 생물막 형성을 억제할 수 있으며, 막을 이용한 수처리 공정에서 막에 발생하는 biofouling을 완화시킬 수 있는 새로운 anti-fouling 처리 기술로서 이러한 QQ 효소의적용 가능성을 보여 주고 있다.

• Restorative Dentistry and Endodontics
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• 제33권6호
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• pp.560-569
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• 2008

본 review 논문의 목적은 통상의 근관치료로 해결되지 않는 persistent periapical lesion의 원인이 되는 주요 세균을 제거하고자 시행한 여러가지 실험을 비교분석하여 과연 (1) Enterococcus faecalis가 근관치료 실패의 주요 원인균인지 (2) 그리고 과연 그렇다면 근관치료에 실패한 증례에서 E. faecalis와 biofilm을 제거할 수 있는 치료 protocol이 있는 것인지를 확인하여 보다 나은 근관치료 성공을 위한 치료 protocol의 확립과 앞으로의 연구방향을 재조명하는 것이다. 지금까지 진행되어온 연구 결과에 대한 객관적인 분석이나 적절한 평가가 이루어지지 않은 가운데 어떤 특정한 연구를 통해 E. faecalis를 제거하는데 유의성있는 효과를 보인다고 알려진 세척액이나 약제를 막연한 기대감을 가지고 실제 임상에 사용하고 있는 실정에서 현재 진료실에서 사용하고 있는 치료 protocol에 대한 검증이 절실한 시점에서 review해 본 결과 현재까지 진행되어 왔던 여러 연구 결과를 통해 확신할 수 있는 것은 치료 protocol에 따라 현재 사용하고 있는 근관세척액이나 근관내 약제만으로도 E. faecalis나 그 biofilm을 대부분 제거할 수 있다는 사실이다. 하지만 이 그 protocol에 따라 근관치료 술식을 충실하게 이행한다 해도 근관치료가 100% 성공한다고 보장할 수는 없다. 물론 세균이 아닌 다른 요소에 의해 근관치료의 실패가 일어난다고도 할 수 있지만 그보다는 결국 체내의 면역반응에 저항하는 세균의 능력에 기인하는 것으로 보인다. 따라서 높은 수준의 치료 성공률을 지속적으로 유지하기 위해서는 위에서 언급된 바와 같은 제대로 된 치료 protocol을 따라 근관치료를 진행하면서 좀더 나은 결과를 얻기 위해 새로운 protocol을 개발하고 정립하는 과정이 계속되어야 한다.

• 미생물학회지
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• 제31권3호
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• pp.255-260
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• 1993

Enterobacter cloacae 를 이용하여 슬라이드 (slide glass) 와 아연도강관조각 (galvanized-iron coupon) 상에 생물막을 형성시킨 후 부착생장세균의 염소에 대한 내성을 측정한 결과 부유생장세균에 비해 각각 14배와 380배의 내성증가를 나타내었다. 또한 입자에 부착된 세균군집의 경우 부유세균군집에 비해 48배로 내성이 증가되었다. 슬라이드와 아연도강관조각을 수돗물에 75일간 접촉하였을때 각각 $4.75 {\times} 10^{4}$ cfu/$cm^{2}$, $1.12 {\times} 10^5 cfu/cm^{2}$의 생물막이 형성되는 것을 관찰하였다. 따라서 수돗물에서의 장내세균과 종속영양세균 입자에 부착 혹은 흡착되거나 응집된 상태로 존재함으로써 잔류염소에 대한 내성을 가지게 되며, 또한 정수과정에서 염소살균을 피하여 비급수계통으로 유입되면 배급수관 내벽에 생물막을 형성하여 잔류업소에 대한 내성을 가지게 됨으로써 배급수계통에서 생장하는 것으로 판단된다.