Jaeeun Kim

Silver-ion-mediated reactive oxygen species generation affecting bactericidal activity.

Silver ions have been widely used as disinfectants that inhibit bacterial growth by inhibiting the essential enzymatic functions of the microorganism via interaction with the thiol-group of l-cysteine. However, silver-ion-mediated perturbation of the bacterial respiratory chain has raised the possibility of reactive oxygen species (ROS) generation. We used bacterial reporter strains specifically responding to superoxide radicals and found that silver-ion-mediated ROS-generation affected bactericidal activity. Almost half the log reduction in Escherichia coli and Staphylococcus aureus populations (model strains for gram negative and positive bacteria, respectively) caused by silver-ion disinfection was attributed to ROS-mediated bactericidal activity. The major form of ROS generated was the superoxide-radical; H(2)O(2) was not induced. Furthermore, silver ions strongly enhanced paraquat-induced oxidative stress, indicating close correlation and synergism between the conventional and ROS-mediated silver toxicity. Our results suggest that further studies in silver-based disinfection systems should consider the oxygen concentration and ROS reaction.

Comparison of the Antimicrobial Effects of Chlorine, Silver Ion, and Tobramycin on Biofilm

ABSTRACT The systematic understanding of how various antimicrobial agents are involved in controlling biofilms is essential in order to establish an effective strategy for biofilm control, since many antimicrobial agents are effective against planktonic cells but are ineffective when they are used against the same bacteria growing in a biofilm state. Three different antimicrobial agents (chlorine, silver, and tobramycin) and three different methods for the measurement of membrane integrity (plate counts, the measurement of respiratory activity with 5-cyano-2,3-ditolyl tetrazolium chloride [CTC] staining, and BacLight Live/Dead staining) were used along with confocal laser scanning microscopy (CLSM) and epifluorescence microscopy to examine the activities of the antimicrobials on biofilms in a comparative way. The three methods of determining the activities of the antimicrobials gave very different results for each antimicrobial agent. Among the three antimicrobials, tobramycin appeared to be the most effective in reducing the respiratory activity of biofilm cells, based upon CTC staining. In contrast, tobramycin-treated biofilm cells maintained their membrane integrity better than chlorine- or silver-treated ones, as evidenced by imaging by both CLSM and epifluorescence microscopy. Combined and sequential treatments with silver and tobramycin showed an enhanced antimicrobial efficiency of more than 200%, while the antimicrobial activity of either chlorine or tobramycin was antagonized when the agents were used in combination. This observation makes sense when the different oxidative reactivities of chlorine, silver, and tobramycin are considered.

Mechanisms of Escherichia coli inactivation by several disinfectants

The objective of this study was to elucidate dominant mechanisms of inactivation, i.e. surface attack versus intracellular attack, during application of common water disinfectants such as ozone, chlorine dioxide, free chlorine and UV irradiation. Escherichia coli was used as a representative microorganism. During cell inactivation, protein release, lipid peroxidation, cell permeability change, damage in intracellular enzyme and morphological change were comparatively examined. For the same level of cell inactivation by chemical disinfectants, cell surface damage was more pronounced with strong oxidant such as ozone while damage in inner cell components was more apparent with weaker oxidant such as free chlorine. Chlorine dioxide showed the inactivation mechanism between these two disinfectants. The results suggest that the mechanism of cell inactivation is primarily related to the reactivity of chemical disinfectant. In contrast to chemical disinfectants, cell inactivation by UV occurred without any changes measurable with the methods employed. Understanding the differences in inactivation mechanisms presented herein is critical to identify rate-limiting steps involved in the inactivation process as well as to develop more effective disinfection strategies.

Furanone derivatives as quorum-sensing antagonists of Pseudomonas aeruginosa

The biofilm formation of Pseudomonas aeruginosa, an opportunistic human pathogen, is developed by cell-to-cell signaling, so-called quorum sensing (QS). To control the biofilm formation, we designed and synthesized new QS inhibitors of P. aeruginosa based on the structure of the previously known QS inhibitor, furanone. Newly synthesized compounds were a series of analogs of (5-oxo-2,5-dihydrofuran-3-yl)methyl alkanoate, and the structures of all six synthesized compounds was confirmed by NMR and GC/MS analyses. These new QS inhibitor candidates could remarkably inhibit both Pseudomonas QS signaling and biofilm formation, which were assayed by using the recombinant reporter system and flow cell confocal microscopy. The degree of QS inhibition by these new inhibitors varied from 20% to 90%. For the profound understanding about inhibition mechanism, we tried to estimate the binding energy between QS receptor, LasR, and our inhibitors from the in silico modeling system. The predicted binding pattern from the modeling system and our experimental data about QS inhibition were in good agreement. From these results, we suggest a new approach to develop the QS inhibitors and biofilm control agents based on structural modeling.

Tolerance of dormant and active cells in Pseudomonas aeruginosa PA01 biofilm to antimicrobial agents

OBJECTIVES The aim of the study was to determine the susceptibility of active and dormant cell populations from Pseudomonas aeruginosa biofilms to non-antibiotic antimicrobial agents such as chlorine, hydrogen peroxide and silver ions in comparison with antibiotics. METHODS Active cells in colony biofilm were differentially labelled by induction of a green fluorescent protein (GFP). Active and dormant cells were sorted in phosphate buffered solution by flow cytometry. Reductions in viability were determined with plate counts. RESULTS The spatial pattern of metabolic activity in colony biofilm was verified, and the active and dormant cells were successfully sorted according to the GFP intensity. Active cells had bigger cell size and higher intracellular density than dormant cells. While dormant cells were more tolerant to tobramycin and silver ions, active cells were more tolerant to chlorine. Metabolically active cells contain denser intracellular components that can react with highly reactive oxidants such as chlorine, thereby reducing the available concentrations of chlorine. In contrast, the concentrations of silver ions and hydrogen peroxide were constant during treatment. Aerobically grown stationary cells were significantly more tolerant to chlorine unlike other antimicrobial agents. CONCLUSIONS Chlorine was more effective in inactivation of metabolically inactive dormant cells and also more effective under anaerobic conditions. The high oxidative reactivity and rapid decay of chlorine might influence the different antimicrobial actions of chlorine compared with antibiotics. This study contributes to understanding the effects of dormancy and the presence of oxygen on the susceptibility of P. aeruginosa biofilm to a wide range of antimicrobial agents.

Ecotope mapping for landscape ecological assessment of habitat and ecosystem

An ecotope (spatial eco-space) map that considers topography and bio-organism-relevant variables emerges as an important basic framework when landscape-scale characteristics for ecosystem management and wildlife conservation are needed. A spatio-geoecological framework based on geographic information systems (GIS) and a vegetation survey were developed for wildlife habitat evaluation of national parks and applied to a representative rugged valley area of Mt. Sorak National Park in Korea. An ecotope map was classified into hundreds of types and dozens of groups by combining biological and geophysical variables. Variables included: forest vegetation type, topographic solar radiation, normalized difference vegetation index (NDVI), elevation, and anthropogenic factors, such as, streams and roads. Layers of GIS variables were produced by field surveys, modeling, satellite images, or digitalization. Vegetation surveys were carried out to identify finer-scale distribution of vegetation types in the rugged valley area. Digital forest vegetation maps from the Forestry Administrator were then modified using the field-surveyed vegetation maps. Topographic solar radiation was predicted with a daily topographic radiation model. The NDVI was calculated from the satellite imagery of a Landsat Thematic Mapper. A digital elevation model (DEM) was used and the other layers were digitized using topographical maps with a scale of 1:25 000. The aim of this study is to determine the geoecological factors relating to the spatial pattern of plant community. It was cleared by the spatial pattern of environmental variables and vegetation characteristics by detrended correspondence analysis using plant species and the environmental variables of each plot. The ordination component value of the first axis shows significant regression to some environmental variables. A case study of habitat evaluation was carried out using the resultant ecotope map. The spatial distribution of potential goral habitat and vegetation characteristics were predicted and the impact of human trails on the neighboring vegetation was also examined for restoration planning. The GIS-based framework developed for wildlife habitat evaluation is useful for natural resource management and human activity control in national parks in Korea.

Inhibition of bacterial adhesion on well ordered comb-like polymer surfaces

The surfaces of comb-like poly(oxyethylene) derivatives with n-alkylsulfonyl side groups were more effective at reducing Pseudomonas aeruginosa adhesion than the surfaces of common materials such as polystyrene, poly(methyl methacrylate), poly(dimethylsiloxane), fluorinated polyacrylate, and glass. When the comb-like poly(oxyethylene) was mixed with polystyrene and poly(methyl methacrylate), the topology and roughness of the surfaces varied according to the mixture compositions. However the surface energies of the mixtures were close to that of the comb-like poly(oxyethylene) in the range of 21-23 mN/m and bacterial adhesion resistances of the mixture surfaces were also comparable to that of the pure comb-like poly(oxyethylene) surface.

Inactivation of Pseudomonas aeruginosa biofilm by dense phase carbon dioxide

Dense phase carbon dioxide (DPCD) is one of the most promising techniques available to control microorganisms as a non-thermal disinfection method. However, no study on the efficiency of biofilm disinfection using DPCD has been reported. The efficiency of DPCD in inactivating Pseudomonas aeruginosa biofilm, which is known to have high antimicrobial resistance, was thus investigated. P. aeruginosa biofilm, which was not immersed in water but was completely wet, was found to be more effectively inactivated by DPCD treatment, achieving a 6-log reduction within 7 min. The inactivation efficiency increased modestly with increasing pressure and temperature. This study also reports that the water-unimmersed condition is one of the most important operating parameters in achieving efficient biofilm control by DPCD treatment. In addition, observations by confocal laser scanning microscopy revealed that DPCD treatment not only inactivated biofilm cells on the glass coupons but also caused detachment of the biofilm following weakening of its structure as a result of the DPCD treatment; this is an added benefit of DPCD treatment.

Effect of the PEG or PMMA micro-patterned surface roughness on bacterial adhesion

In this paper, we investigate the bacterial adhesion behaviors of microstructured PEG and PMMA surfaces. Microstructures on PEG and PMMA surface were fabricated using capillary lithography mediated by ultraviolet (UV) exposure for PEG and heating for PMMA to control the roughness of surface in terms of manipulating size and shape of patterns