• Journal of Periodontal and Implant Science
• /
• v.49 no.3
• /
• pp.193-204
• /
• 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
• /
• v.39 no.5
• /
• pp.417-429
• /
• 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.

• Journal of dental hygiene science
• /
• v.18 no.2
• /
• pp.69-75
• /
• 2018

The purpose of our study is to compare the adhesion and biofilm formation abilities of isolates from water discharged from dental unit waterlines (DUWLs). Bacteria were isolated from a total of 15 DUWLs. Twelve isolates were selected for the experiment. To confirm the adhesion ability of the isolates, each isolate was attached to a glass coverslip using a 12-well plate. Plates were incubated at $26^{\circ}C$ for 7 days, and the degree of adhesion of each isolate was scored. To verify the biofilm formation ability of each isolate, biofilms were allowed to form on a 96-well polystyrene flat-bottom microtiter plate. The biofilm accumulations of all isolates formed at $26^{\circ}C$ for 7 days were identified and compared. A total of 56 strains were isolated from 15 water samples including 12 genera and 31 species. Of the 56 isolates, 12 isolates were selected according to the genus and used in the experiment. Sphingomonas echinoides, Methylobacterium aquaticum, and Cupriavidus pauculus had the highest adhesion ability scores of +3 among 12 isolates. Among these three isolates, the biofilm accumulation of C. pauculus was the highest and that of S. echinoides was the third-most abundant. The lowest biofilm accumulations were identified in Microbacterium testaceum and M. aquaticum. Most isolates with high adhesion ability also exhibited high biofilm formation ability. Analysis of adhesion and biofilm formation of the isolates from DUWLs can provide useful information to understand the mechanism of DUWL biofilm formation and development.

• Journal of Korean Society of Water and Wastewater
• /
• v.27 no.6
• /
• pp.689-701
• /
• 2013

Bacterial growth and corresponding consumption of carbon and phosphorus were examined in which tap water samples containing a very low concentration of free chlorine were supplemented with organic carbon and/or phosphorus. The experiments were performed in a fed-batch mode under a controlled temperature of $20^{\circ}C$. In the phosphorus alone-added water, there was no significant increase in bacterial numbers measured as heterotrophic plate count (HPC) in the bulk water. However, bacterial growth was stimulated by the addition of carbon (e.g., bulk HPC levels increased to $10^3CFU/mL$) and further stimulated by the combined addition of carbon and phosphorus (e.g., bulk HPC to $10^5CFU/mL$). The same effects were observed in biofilm HPC and biomass formed on polyethylene (PE) slide surfaces. In the water where organic carbon and phosphorus were added together, the highest biofilm HPC and biomass (measured as extracellular polymeric substance components) densities were observed which were $7.6{\times}10^5CFU/cm^2$ and $5.3{\mu}g/cm^2$, respectively. In addition to the bacterial growth, additions of organic carbon and/or phosphorus resulted in different bacterial carbon-to-phosphorus (C/P) consumption ratios. Compared to a typical bacterial C/P consumption ratio of 100:1, a higher C/P ratio (590:1) occurred in the carbon alone-added water, while a lower ratio (40:1) in phosphorus alone-added water. Comparative value (80:1) of C/P ratio was also observed in the water where organic carbon and phosphorus were added together. At the given experimental conditions, bacterial growth was deemed to be more sensitive to microbially available organic carbon than phosphorus. The effect of phosphorus addition, which resulted in a lower C/P consumption ratio, seemed to be tightly associated with the presence of microbially available organic carbon. These results suggested that the control of extrinsic carbon influx seemed to be more important to minimize bacterial regrowth in drinking water system, since even low content of phosphorus naturally occurring in drinking water was enough to allow a bacterial growth.

• Corrosion Science and Technology
• /
• v.18 no.6
• /
• pp.221-227
• /
• 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
• /
• v.15 no.5
• /
• pp.938-945
• /
• 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.

• Microbiology and Biotechnology Letters
• /
• v.32 no.1
• /
• pp.1-10
• /
• 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.

• Journal of Korean Society of Water and Wastewater
• /
• v.20 no.3
• /
• pp.431-441
• /
• 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
• /
• v.31 no.4
• /
• pp.661-668
• /
• 2001

Anti-P. gingivalis immune sera were obtained from mice immunized with either P. gingivalis alone, or F. nucleaturm followed by P. gingivalis. Two groups of immune sera were examined for binding capacity to P. gingivalis biofilm by confocal laser scanning microscope, Antibody avidity index was also determined for each immune sera. The results indicated that prior immunization of mice with F. nucleaturm impaired P. gingivalis-specific immune sera in binding capacity to biofilm and antibody avidity to P. gingivalis. Elevated antibody responses in patients with destructive periodontal disease has often been related to suboptimal level of protective antibody $(opsonophagocytosis)^{1-3)}$ while post-immune sera obtained with experimental animals using a single periodontal pathogen demonstrated satisfactory levels of protective function against the homologous bacterial $challenge^{4,5)}$.The reason is unclear why elevated IgG responses in periodontal patients to periodontal pathogens do not necessarily reflect their protective function. Such an immune deviation might be derived from the fact that destructive periodontal disease is cumulative result of immunopathologic processes responding to an array of different colonizing microorganisms sequentially infecting in the subgingival environmental niche. Fusobacterium nucleaturm is one of the key pathogens in gingivitis, in the transitional phase of conversion of gingivitis into destructive periodontitk, and in adult $periodontitis^{6-8)}$. It also plays a central role in coaggregation with other important microbial species in subgingival $area^{6,9,10)}$ as well as in $biofilm^{11)}$, especially with Porphyromonas gingjvalis in synergism of virulence in human periodontal disease or in animal $models^{12-14)}$. This organism has also been reported to have immune modulating activity for secondary immune response to Actinobacillus $actinomycetemcomitans^{15)}$. It is presumed that sequential colonization and intermicrobial coaggregation between intermediate and late colonizers could potentially modulate the immune responses and development of specific T cell phenotypes in periodontal lesions. We have recently demonstrated the skewed polarization of P. gingivalis-specific helper T cell clones in mice immunized with F. nucleaturm followed by P. $gingivalis.^{16)}$. Consequently F. nucleaturm may initially prime the immune cells and modify their responses to the successive organism, P. gingivalis. This could explain why one frequently observes non-protective serum antibodies to P. gingivalis in periodontal patients in contrast with those obtained from animals that were immunized with $P.gingivalis\;alone^{17)}$. The present study was performed to investigate the immune modulating effect of F. nucleatum on serum binding to experimental biofilms and the avidity of anti-P. gingivalis antibody.

• Journal of Periodontal and Implant Science
• /
• v.48 no.3
• /
• pp.164-173
• /
• 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.