• Biomedical Science Letters
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• v.9 no.3
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• pp.139-144
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• 2003

Since water borne infection causes acute diseases and results in spread of diseases by secondary infection, the prevention is very important. Therefore, it is necessary to have a method that is rapid and effective to monitor pathogenic bacteria in drinking water. In this study, we employed a systematic method, Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) analysis, to develop an effective monitoring system for possible bacterial contaminants in drinking water. For this purpose, PCR primers were derived from 992 bp region of the 16s rRNA gene that is highly conserved through the different species of prokaryotes. To test whether the PCR primers designed are indeed useful for detecting all the possible microbial contaminants in the water, the primers were used to amplify 16s rRNA regions of different microbial water-borne pathogens such as E. coli, Salmonella, Yersinia, Listeria, and Staphylococcus. As expected, all of tested microorganisms amplified expected size of PCR products indicating designed PCR primers for 16s rRNA indeed can be useful to amplify all different microbial water-borne pathogens in the water. Furthermore, to test whether these 16s rRNA based PCR primers can detect bacterial populations present in the water, water samples taken from diverse sources, such as river, tap, and sewage, were used for amplification. PCR products were for then subjected for cloning into a T-vector to generate a library containing 16s rRNA sequences from various bacteria. With cloned PCR products, RFLP analysis was done using PCR products digested with restriction enzyme such as Hae III to obtain species-specific RFLP profiles. After PCR-RFLP, the bacterial clones which showed the same RFLP profiles were regarded as the same ones, and the clones which showed distinctive RFLP profiles were subsequently subjected for sequence analysis for species identification. By this PCR-RFLP analysis, we were able to reveal diverse populations of bacteria living in water. In brief, in unsterilized natural river water, over 60 different species of bacteria were found. On the other hand, no PCR products were detected in drinking tap-water. The results from this study clearly indicate that the PCR-RFLP-sequence analysis can be a useful method for monitoring diverse, perhaps pathogenic bacteria contaminated in water in a rapid fashion.

• Journal of Microbiology and Biotechnology
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• v.26 no.1
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• pp.120-129
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• 2016

An industrial complex in Wonju, contaminated with trichloroethene (TCE), was one of the most problematic sites in Korea. Despite repeated remedial trials for decades, chlorinated ethenes remained as sources of down-gradient groundwater contamination. Recent efforts were being made to remove the contaminants of the area, but knowledge of the indigenous microbial communities and their dechlorination abilities were unknown. Thus, the objectives of the present study were (i) to evaluate the dechlorination abilities of indigenous microbes at the contaminated site, (ii) to characterize which microbes and reductive dehalogenase genes were responsible for the dechlorination reactions, and (iii) to develop a PCE-to-ethene dechlorinating microbial consortium. An enrichment culture that dechlorinates PCE to ethene was obtained from Wonju stream, nearby a trichloroethene (TCE)-contaminated industrial complex. The community profiling revealed that known organohalide-respiring microbes, such as Geobacter, Desulfuromonas, and Dehalococcoides grew during the incubation with chlorinated ethenes. Although Chloroflexi populations (i.e., Longilinea and Bellilinea) were the most enriched in the sediment microcosms, those were not found in the transfer cultures. Based upon the results from pyrosequencing of 16S rRNA gene amplicons and qPCR using TaqMan chemistry, close relatives of Dehalococcoides mccartyi strains FL2 and GT seemed to be dominant and responsible for the complete detoxification of chlorinated ethenes in the transfer cultures. This study also demonstrated that the contaminated site harbors indigenous microbes that can convert PCE to ethene, and the developed consortium can be an important resource for future bioremediation efforts.

• KSBB Journal
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• v.13 no.5
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• pp.469-476
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• 1998

Among 31 water-born microbial strains isolated from various sites in Korea, strain DJ-4 was selected as a test organism for toxicity measurements in that its growth was completely inhibited by the presence of 668.4 mg/L of chloroform and 297.5 mg/L of toluene in the liquid LB medium whereas others did not. It was observed that lag periods and specific growth rates of DJ-4 batch vial cultures were prolonged and decreased, respectively, by phenol, benzene, toluene, ethylbenzene, p-xylene, perchloroethylene, trichloroethylene, and chloroform at the concentrations between 3.6 and 417.8 mg/L. There changes were found to be linear with respect to the concentrations of the toxic compounds. From the first-order regression equations, 50% effective concentrations (EC50${\mu}$ for concentrations of toxic compounds causing 50% decrease of specific growth rates and EC50lag for 50% increase of length of lag periods) were calculated for each compounds. By comparing DJ-4 EC50${\mu}$ values with Daphnia LC50's from a literature for benzene, ethylbenzene, toluene, and trichloroethlyene, it was concluded that microbial specific growth could be a new, fast, and reliable parameter for toxicity tests.

• Molecular & Cellular Toxicology
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• v.1 no.3
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• pp.189-195
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• 2005

Industrial development has increase consumption of crude oil and environmental pollution. A large number of microbial lipolytic enzymes have been identified and characterized to date. To development for a new lipase with catalytic activity in degradation of crude oil as a microbial enzyme, Acinetobactor sp. B2 was isolated from soil samples that were contaminated with oil in Daejon area. Acinetobactor sp. B2 showed high resistance up to 10 mg/mL unit to heavy metals such as Ba, Li, Al, Cr, Pb and Mn. Optimal growth condition of Acinetobactor sp. B2 was confirmed $30^{\circ}C$. Lipase was purified from the supernatant by Acinetobactor sp. B2. Its molecular mass was determined to the 60 kDa and the optimal activity was shown at $40^{\circ}C$ and pH 10. The activation energies for the hydrolysis of p-nitrophenyl palmitate were determined to be 2.7 kcal/mol in the temperature range 4 to $37^{\circ}C$. The enzyme was unstable at temperatures higher than $60^{\circ}C$. The Michaelis constant $(K_{m})\;and\;V_{max}$ for p-nitrophenyl palmitate were $21.8{\mu}M\;and\;270.3{\mu}M\;min^{-1}mg\;of\;protein^{-1}$, respectively. The enzyme was strongly inhibited by $Cd{2+},\;Co^{2+},\;Fe^{2+},\;Hg^{2+},\;EDTA$, 2-Mercaptoethalol. From these results, we suggested that lipase purified from Acinetobactor sp. B2 should be able to be used as a new enzyme for degradation of crude oil, one of the environmental contaminants.

• Environmental Engineering Research
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• v.21 no.2
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• pp.164-170
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• 2016

Since most of the existing wastewater treatment options lack the ability to treat micro-contaminants, the increased use of pharmaceuticals and personal care products (PPCPs) and release as human waste have become a serious concern in recent years. Constructed wetlands (CWs) are a low-cost technology for wastewater treatment, however, its performance in term of PPCPs removal has not yet been fully investigated. This study aimed to characterize the removal factors and efficiency of acetaminophen (ACT) removal by CWs. The results revealed the decreased concentrations of ACT with increasing hydraulic retention times (HRT) of 0, 3, 5 days. The contribution of removal factors was found to be varied with initial ACT concentration. At the low ACT concentration (i.e. 1 ppb), plant uptake was the dominant, followed by microbial and photolytic removal. In contrast, at the high ACT concentration (i.e. 100 ppb), microbial and photolytic removal were found as dominant factors. On the other hand, hydrogen peroxide ($H_2O_2$) concentration was found at higher level in the plant shoot than in the root probably due to occurrence of the Fenton reaction resulting in PPCPs removal.

• Journal of Environmental Health Sciences
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• v.34 no.1
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• pp.86-94
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• 2008

Lake Shiwha, an artificial lake located near metropolitan Seoul, offers a unique water environment and has been suspected to have high levels of chemical and microbiological contaminations. Lake Shiwha was originally connected to the sea but currently has four major surface water inputs from agricultural, municipal, industrial areas and in addition an occasional inflow from the sea. The objectives of this study are to investigate the relative contribution of microbial contaminants from each of the inflowing surface waters and to identify appropriate microbial indicator organisms in this unique water environment. We measured the levels of microbial contaminations in the four inflowing surface waters. A number of microbial indicator organisms including total coliform (TC), fecal coliform (FC), E. coli, Enterococci, somatic and male-specific coliphages were analyzed. Bacterial indicator microorganisms were detected and quantified by the $Colilert^{(R)},\;Enterolert^{(R)}$ kit. Surface water (50 l) was sampled by $ViroCap^{TM}\;5"$ cartridge filters and analyzed by the single agar layer method for detecting coliphages. The concentrations of TC, FC, E. coli, and Enterococci were 1543 CFU/100 ml${\sim}1.99{\times}10^6$ CFU/100 ml, 0 CFU/100 ml${\sim}202$ CFU/100ml, 0 CFU/100 ml${\sim}1.80{\sim}10^5$ CFU/100ml, 74 CFU/100 ml${\sim}3408$ CFU/100 ml, respectively. The male-specific and somatic coliphages were detected in three different inflowing surface waters. Isolated E. coli and Enterococci strains were further analyzed by 16s rDNA amplification and subsequent phylogenetic analysis from Jungwang-chun, Ansan-chun, Banwol-chun and penstock of inflowing surface water. Our results indicated that the concentrations of different fecal indicator microorganisms might not be highly correlated with each other. Multiple microbial indicator organisms should be used for monitoring microbial contamination and microbial source tracking methods.

• Journal of Soil and Groundwater Environment
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• v.18 no.3
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• pp.73-81
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• 2013

Various remediation methods have been applied to clean soils contaminated with pollutants. They remove contaminants from the soils by utilizing physicochemical, biological, and thermal processes and can satisfy soil remediation standards within a limited time; however, they also have an effect on the biological functions of soils by changing soil properties. In this study, changes of the biological properties of soils before and after treatment with three frequently used remediation methods-soil washing, land farming, and thermal desorption-were monitored to investigate the effects of remediation methods on soil biological functions. Total microbial number and soil enzyme activities, germination rate and growth of Brassica juncea, biomass change of Eisenia andrei were examined the effects on soil microorganisms, plant, and soil organisms, respectively. After soil washing, the germination rate of Brassica juncea increased but the above-ground growth and total microbial number decreased. Dehydrogenase activity, germination rate and above-ground growth increased in both land farming and thermal desorption treated soil. Although the growth of Eisenia andrei in thermal desorption treated soil was higher than any other treatment, it was still lower than that in non-contaminated soil. These results show that the remediation processes used to clean contaminated soil also affect soil biological functions. To utilize the cleaned soil for healthy and more value-added purposes, soil improvement and process development are needed.

• Environmental Engineering Research
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• v.12 no.2
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• pp.37-45
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• 2007

Phytoremediation has been used effectively for the biodegradation of oil-based contaminants, including diesel, by the stimulation of soil microbes near plant roots (rhizosphere). However, the technique has rarely been assessed for itsinfluence on soil microbial properties such as population, community structure, and diversity. In this study, the removal efficiency and characteristics of rhizobacteria for phytoremediation of diesel-contaminated soils were assessed using barnyard grass (Echinochloa crusgalli). The concentration of spiked diesel for treatments was around $6000\;mg\;kg^{-1}$. Diesel removal efficiencies reached 100% in rhizosphere soils, 76% in planted bulk soils, and 62% in unplanted bulk soils after 3weeks stabilization and 2 months growth(control, no microbial activity: 32%). The highest populations of culturable soil bacteria ($5.89{\times}10^8$ per g soil) and culturable hydrocarbon-degraders($5.65{\times}10^6$ per g soil) were found in diesel-contaminated rhizosphere soil, also yielding the highest microbial dehydrogenase. This suggests that the populations of soil bacteria, including hydrocarbon-degraders, were significantly increased by a synergistic rhizosphere + diesel effect. The diesel treatment alone resulted in negative population growth. In addition, we investigated the bacterial community structures of each soil sample based on DGGE (Denaturing Gel Gradient Electrophoresis) band patterns. Bacterial community structure was most influenced by the presence of diesel contamination (76.92% dissimilarity to the control) and by a diesel + rhizosphere treatment (65.62% dissimilarity), and least influenced by the rhizosphere treatment alone (48.15% dissimilarity). Based on the number of distinct DGGE bands, the bacterial diversity decreased with diesel treatment, but kept constant in the rhizosphere treatment. The rhizosphere thus positively influenced bacterial population density in diesel-contaminated soil, resulting in high removal efficiency of diesel.

• KSBB Journal
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• v.21 no.6 s.101
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• pp.428-432
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• 2006

This study investigated the effect of sulfate source on removal efficiency in electrokinetic bioremediation which needs sulfate to degrade contaminants by an applied microorganism. The representative contaminant and the applied microorganism were phenanthrene and Sphingomonas sp. 3Y, respectively. When magnesium sulfate was used, the magnesium ion combined with hydroxyl ion electrically-generated at cathode to cause the decrease of electrolyte pH, and then the microbial activity was inhibited by that. When ammonium sulfate and disodium sulfate were used to solve the pH control problem, the pH values of electrolyte and soil solution were maintained neutrally, and also the high microbial activity was observed. With the former sulfate source, however, ammonium retarded the phenanthrene degradation, and so the removal efficiency decreased to 12.0% rather than 21.8% with magnesium sulfate. On the other hand, the latter improved the removal efficiency to 27.2%. This difference of removal efficiency would be outstanding for an elongated treatment period.

• Environmental Engineering Research
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• v.20 no.2
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• pp.149-154
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• 2015

In recent years, immobilization agents were introduced into organic contaminated soil remediation and more and more materials were screened and used as the immobilizing carrier. However, effect of the decomposition of the immobilizing carrier on the bioremediation was rarely concerned. Therefore, the decomposition experiment of immobilizing carrier -corncob was carried out in the lab with the efficient degradation fungi - Mucor mucedo (MU) existing, and polycyclic aromatic hydrocarbons (PAHs) residues E4/E6 of the dissolved organic matter and microbial diversity during the decomposition process were studied. The results showed that: a) during the decomposition, the degradation of pyrene (Pyr) was mainly in the first 28 d in which the content of extractable Pyr decreased rapidly and the highest decrease was in the treatment with only MU added. b) Anslysis of E4/E6 changes showed that rich microorganisms could promote aromatization and condensation of humus. c) From the diversity index analysis it can also be seen that there is no significant difference in effects of PAHs on the uniformity of microorganisms. These results will not only be useful to have a better understanding of the bioavailability of contaminants adsorbed to biodegradable carriers in PAHs contaminated soil remediation, but also be helpful to perfect the principle of immobilized microbial technique.