• Journal of Periodontal and Implant Science
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• 제49권3호
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• pp.193-204
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• 2019

Purpose: The reaction of cells to a titanium implant depends on the surface characteristics of the implant which are affected by decontamination. The aim of this study was to evaluate the cytocompatibility of titanium disks treated with various decontamination methods, using salivary bacterial contamination with dental pellicle formation as an in vitro model. Methods: Sand-blasted and acid-etched (SA) titanium disks were used. Three control groups (pristine SA disks [SA group]; salivary pellicle-coated SA disks [pellicle group]; and biofilm-coated, untreated SA disks [NT group]) were not subjected to any decontamination treatments. Decontamination of the biofilm-coated disks was performed by 14 methods, including ultrasonic instruments, rotating instruments, an air-powder abrasive system, a laser, and chemical agents. MG63 cells were cultured in the presence of the treated disks. Cell proliferation assays were performed on days 2 and 5 of cell culture, and cell morphology was analyzed by immunofluorescence and scanning electron microscopy (SEM). A vascular endothelial growth factor (VEGF) assay was performed on day 5 of culture. Results: The cell proliferation assay revealed that all decontaminated disks, except for the 2 groups treated using a plastic tip, showed significantly less cell proliferation than the SA group. The immunofluorescence and SEM analyses revealed that most groups showed comparable cell density, with the exception of the NT group, in which the cell density was lower and bacterial residue was observed. Furthermore, the cells grown with tetracycline-treated titanium disks showed significantly lower VEGF production than those in the SA group. Conclusions: None of the decontamination methods resulted in cytocompatibility similar to that of pristine SA titanium. However, many methods caused improvement in the biocompatibility of the titanium disks in comparison with the biofilm-coated, untreated titanium disks. This suggests that decontamination is indispensable for the treatment of peri-implantitis, even if the original biocompatibility cannot be restored.

• The Plant Pathology Journal
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• 제39권5호
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• pp.417-429
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• 2023

Ralstonia solanacearum species complex (RSSC) is a soil borne plant pathogen causing bacterial wilt on various important crops, including Solanaceae plants. The bacterial pathogens within the RSSC produce exopolysaccharide (EPS), a highly complicated nitrogencontaining heteropolymeric polysaccharide, as a major virulence factor. However, the biosynthetic pathway of the EPS in the RSSC has not been fully characterized. To identify genes in EPS production beyond the EPS biosynthetic gene operon, we selected the EPS-defective mutants of R. pseudosolanacearum strain SL341 from Tn5-inserted mutant pool. Among several EPSdefective mutants, we identified a mutant, SL341P4, with a Tn5-insertion in a gene encoding a putative NDP-sugar epimerase, a putative membrane protein with sugar-modifying moiety, in a reverse orientation to EPS biosynthesis gene cluster. This protein showed similar to other NDP-sugar epimerases involved in EPS biosynthesis in many phytopathogens. Mutation of the NDP-sugar epimerase gene reduced EPS production and biofilm formation in R. pseudosolanacearum. Additionally, the SL341P4 mutant exhibited reduced disease severity and incidence of bacterial wilt in tomato plants compared to the wild-type SL341 without alteration of bacterial multiplication. These results indicate that the NDP-sugar epimerase gene is required for EPS production and bacterial virulence in R. pseudosolanacearum.

• 치위생과학회지
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• 제18권2호
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• pp.69-75
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• 2018

우리 연구의 목적은 DUWL에서 배출되는 물에서 분리한 균주의 부착 능력과 바이오필름 형성 능력을 확인하고 두 능력 사이 관계를 확인하는 것이다. DUWL로부터 분리한 12균주를 실험에 사용하였다. 각 균주의 부착 능력을 확인하기 위해, 12-well plates의 각 well에 멸균된 glass coverslip, R2A 액체 배지, 그리고 $1{\times}10^8CFU/ml$의 농도로 조정된 세균 현탁액을 넣고 $26^{\circ}C$ 배양기에서 7일 동안 배양하였다. 배양 후, glass coverslip에 부착한 정도에 따라 1~3점으로 점수를 부여하였다. 분리 균주의 바이오필름 형성 능력을 확인하기 위해, 96-well polystyrene flat-bottom microtiter plate에 R2A 액체 배지와 세균 현탁액을 넣고 $26^{\circ}C$에서 7일 동안 배양하였다. 배양 후, plate에 형성된 바이오필름은 R2A 액체 배지에 현탁했고, 현탁액을 R2A 고체 배지에 도말하였다. $26^{\circ}C$에서 7일 배양한 후 세균의 집락을 계수하고 CFU/ml를 계산하였다. DUWL로부터 총 56균주가 분리되었으며, 12속과 31종을 포함하였다. 실험에는 속당 1균주씩 선택하여 총 12균주가 사용되었다. 12균주 중에 S. echinoides, M. aquaticum, C. pauculus의 부착 능력 점수는 +3으로 가장 높았다. 바이오필름 축적량은 C. pauculus가 가장 많았고, M. testaceum이 가장 적었다. 대부분의 부착 능력이 높은 균주는 바이오필름 형성 능력 또한 높았다. 본 연구의 결과는 DUWL 바이오필름의 형성 기전을 파악하는데 도움을 주며 나아가 바이오필름 형성을 억제하는 방법의 개발에 기본적인 정보를 제공할 수 있을 것이다.

• 상하수도학회지
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• 제27권6호
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• pp.689-701
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• 2013

잔류염소 농도가 상당히 낮은 수돗물에서 유기탄소(organic carbon)와 인(phosphorus)의 증가가 세균 생장에 미치는 영향과 세균에 의한 유기탄소와 인의 소모를 fed-batch 실험조건($20^{\circ}C$ 수온)에서 조사하였다. 수돗물에서 단지 인의 증가만으로는 부유성 세균의 현저한 수적 증가는 나타나지 않았다. 그러나 유기탄소의 증가는 부유성 세균을 $10^3CFU/mL$ 수준까지 증가시켰으며, 특히 유기탄소와 함께 동반된 인의 증가는 부유성 세균을 $10^5CFU/mL$ 수준까지 증가시켰다. 이러한 효과들은 polyethylene (PE) slide 표면에 형성된 생물막 세균과 생물량 측정에서도 동일하게 나타났다. 유기탄소와 함께 인 농도가 높은 수돗물에서 PE slide 표면에 형성된 생물막 세균과 세포외 중합체(extracellular polymeric substance) 구성 성분으로 측정된 생물량은 각각 $7.6{\times}10^5CFU/cm^2$와 $5.3{\mu}g/cm^2$로 가장 높았다. 세균 생장과 더불어, 수돗물에서 유기탄소와 인의 증가는 세균에 의한 탄소와 인의 이용 패턴에 영향을 미치는 것으로 나타났다. 전형적인 세균의 C/P 소모비율(100:1)과 비교했을 때, 상대적으로 높은 C/P 소모비율(590:1)은 유기탄소 농도가 높은 수돗물에서, 그리고 상대적으로 낮은 C/P 소모비율(40:1)은 인 농도가 높은 수돗물에서 관찰되었다. 또한 유기탄소와 함께 인 농도가 높은 수돗물에서도 상대적으로 낮은 C/P 소모비율(80:1)이 관찰되었다. 주어진 실험조건에서는 수돗물과 생물막내 세균 생장이 인의 증가보다는 유기탄소의 증가에 더욱 민감하게 반응하는 것으로 나타났다. 수돗물에서 인의 증가는 세균의 낮은 C/P 소모비율을 가져오지만, 세균 생장에 미치는 인의 영향은 세균이 쉽게 이용할 수 있는 유기탄소의 존재와 밀접하게 관련되어 있는 것으로 보인다. 그러므로 수돗물에 자연적으로 존재하는 낮은 농도의 인만으로도 세균 생장을 위해 필요한 인 요구량을 만족시킬 수 있기 때문에, 수돗물에서 세균 생장의 최소화를 위해서는 인보다는 유기탄소의 외부적인 유입을 제어하는 것이 더욱 중요한 것으로 판단된다.

• Corrosion Science and Technology
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• 제18권6호
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• pp.221-227
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• 2019

During the operation, maintenance and decommissioning of nuclear power plant radioactive contaminated waste is produced. This waste is stored in an underground repository 60-100 meters below the surface. The metallic portion of this waste comprises mostly carbon and stainless steel. A long-term field exposure showed high corrosion rates, general corrosion up to 29 ㎛ a^-1 and localized corrosion even higher. High corrosion rate is possible if microbes produce corrosive products, or alter the local microenvironment to favor corrosion. The bacterial and archaeal composition of biofilm formed on the surface of carbon steel was studied using 16S rRNA gene targeting sequencing, followed by phylogenetic analyses of the microbial community. The functional potential of the microbial communities in biofilm was studied by functional gene targeting quantitative PCR. The corrosion rate was calculated from weight loss measurements and the deposits on the surfaces were analyzed with SEM/EDS and XRD. Our results demonstrate that microbial diversity on the surface of carbon steel and their functionality is vast. Our results suggest that in these nutrient poor conditions the role of methanogenic archaea in corrosive biofilm, in addition to sulphate reducing bacteria, could be greater than previously suspected.

• Journal of Microbiology and Biotechnology
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• 제15권5호
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• pp.938-945
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• 2005

For elucidating excessive growth of biofilm that subsequently leads to the clogging problem in a small town's sewer lines of Wisconsin, the FISH method was employed. At the beginning of the simulated experiments, ${\beta}$-subclass proteobacteria prevailed in runs fed with industrial wastewater, while ${\gamma}$-subclass proteobacteria dominated in runs with domestic wastewater. However, the bacterial community structure changed significantly over six weeks; Cytophaga-Flavobacterium (CF)­group bacteria dominated in most runs fed with the small town's wastewater regardless of their source, while CF-group decreased strongly in run fed with domestic sewage from another city (Madison). It was also microscopically confirmed that most of those clogging materials was toilet tissue, which in turn may lead to vigorous growth of cellulose-degrading CF-group bacteria. This dominant presence of CF-group bacteria in the small town's sewer indicates that the main constituent of biofilm, toilet tissue (cellulose) in sewage, might have induced the unique pattern of their microbial community structure. Therefore, it suggests that molecular technique is useful for monitoring the clogging problems in sewer lines.

• 한국미생물·생명공학회지
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• 제32권1호
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• pp.1-10
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• 2004

Many bacteria monitor their population density and control the expression of specialized gene sets in response to bacterial cell density based on a mechanism referred to as quorum sensing. In all cases, quorum sensing involves the production and detection of extracellular signaling molecules, auto inducers, as which Gram-negative and Gram-positive bacteria use most prevalently acylated homoserine lactones and processed oligo-peptides, respectively. Through quorum-sensing communication circuits, bacteria regulate a diverse array of physiological functions, including virulence, symbiosis, competence, conjugation, antibiotic production, motility, sporulation, and biofilm formation. Many pathogens have evolved quorum-sensing mechanisms to mount population-density-dependent attacks to over-whelm the defense responses of plants, animals, and humans. Since these AHL-mediated signaling mechanisms are widespread and highly conserved in many pathogenic bacteria, the disruption of quorum-sensing system might be an attractive target for novel anti-infective therapy. To control AHL-mediated pathogenicity, several promising strategies to disrupt bacterial quorum sensing have been reported, and several chemicals and enzymes have been also investigated for years. These studies indicate that anti-quorum sensing strategies could be developed as possible alternatives of antibiotics.

• 상하수도학회지
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• 제20권3호
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• pp.431-441
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• 2006

This study investigated the physiological characteristics of biofilm microorganisms formed onto the different drinking water distribution pipe surfaces. The simulated drinking water distribution pipe system which had several PVC, STS 304, and GS coupons was operated at flow velocity of 0.08 m/sec (Re 1,950) and 0.28 m/sec (Re 7,300), respectively. At velocity of 0.08 m/sec, the number of viable heterotrophic bacteria in the biofilm over the 3 months of operation averaged $3.3{\times}10^4$, $8.7{\times}10^4$, and $7.2{\times}10^3CFU/cm^2$ for PVC, STS, and GS surfaces, respectively. The number of attached heterotrophic bacteria averaged $1.4{\times}10^3$, $5.6{\times}10^2$, and $6.5{\times}10^2CFU/cm^2$ on PVC, STS, and GS surfaces at the system with relatively high flow velocity of 0.28m/sec. The changes of physiological profile of biofilm-forming microorganisms were characterized by community-level assay that utilized the Biolog GN microplates. Biofilms that formed on different pipe surfaces displayed distinctive patterns of community-level physiological profile (CLPP), which reflected the metabolic preference for different carbon sources and/or the utilization of these carbon sources to varying degrees. The CLPP patterns have shown that the metabolic potential of a biofilm community was different depending on the pipe material. The effect of the pipe material was also characterized differently by operation condition such as flow rate. At flow velocity of 0.08 m/sec, the metabolic potential of biofilm microorganisms on GS surface showed lower levels than PVC and STS biofilms. For biofilms on pipe material surfaces exposed to water flowing at 0.28 m/sec, the metabolic potential was in order of PVC>GS>STS. Generally, the levels of the bacterial biofilm's metabolic potentials were shown to be notably higher on pipe surfaces exposed to water at 0.08 m/sec when compared to those on pipe surfaces exposed to water at 0.28 m/sec.

• Journal of Periodontal and Implant Science
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• 제31권4호
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• pp.661-668
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• 2001

P. gingivalis를 단독면역하거나 또는 Fusobacterium nucleatum 선면역 후 P. gingivalis 항혈청을 각각 얻어냈다. 두 종류의 항혈청이 P. gingivalis biofilm을 침투해 들어가는 능력을 confocal laser scanning microscope를 이용하여 비교 감증하였다. 항혈청의 P. gingivalis에 대한 avidity index도 측정하였다. 결과적으로 F. nucleatum의 선면역은 P. gingivalis 특이 항혈청에 대해 세균성 biofilm의 침투능력을 저하시키고, 동일한 세균에 대한 avidity도 감소시켰다.

• Journal of Periodontal and Implant Science
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• 제48권3호
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• pp.164-173
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• 2018

Purpose: The aim of this study was to evaluate the antimicrobial effect of a newly devised toothbrush with light-emitting diodes (LEDs) on Porphyromonas gingivalis attached to sandblasted and acid-etched titanium surfaces. Methods: The study included a control group, a commercial photodynamic therapy (PDT) group, and 3 test groups (B, BL, and BLE). The disks in the PDT group were placed in methylene blue and then irradiated with a diode laser. The B disks were only brushed, the BL disks were brushed with an LED toothbrush, and the BLE disks were placed into erythrosine and then brushed with an LED toothbrush. After the different treatments, bacteria were detached from the disks and spread on selective agar. The number of viable bacteria and percentage of bacterial reduction were determined from colony counts. Scanning electron microscopy was performed to visualize bacterial alterations. Results: The number of viable bacteria in the BLE group was significantly lower than that in the other groups (P<0.05). Scanning electron microscopy showed that bacterial cell walls were intact in the control and B groups, but changed after commercial PDT and LED exposure. Conclusions: The findings suggest that an LED toothbrush with erythrosine treatment was more effective than a commercial PDT kit in reducing the number of P. gingivalis cells attached to surface-modified titanium in vitro.