Daejeon, South Korea
Daejeon, South Korea

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Nam S.,Chungnam National University | Kim M.-J.,Water Analysis and Research Center | Park C.,Catholic University of Korea | Park J.-G.,Daegu University | And 2 more authors.
International Journal of Hygiene and Environmental Health | Year: 2013

The distribution characteristics of Enterococcus spp., which are indicators of fecal pollution, were investigated at 33 sites within the 3 major water systems of Korea. Enterococci were detected at concentrations ranging from 1 to 37 CFU/100. mL in 41 of 132 samples (31.1%) from the 3 major water systems. The overall average detected concentration was 1.2 CFU/100. mL, while the average concentration for all detection sites was 5.3 CFU/100. mL. After optimized multiplex polymerase chain reaction (PCR) was performed with newly developed VanA, VanB, VanC-1, and VanC-2/3 primers, concentrations of vancomycin-resistant Enterococcus spp. (VRE) ranging from 1 to 23 CFU/100. mL were detected in 17 of 132 samples (12.9%). Of 216 individual enterococcal colonies, 64 (29.6%) displayed the VanC genotype. The results of a susceptibility test to vancomycin showed that the range of the minimal inhibitory concentration (MIC), an indicator of bacterial resistance, was 4 to 24 μg/mL, with the average MIC at 9.2 ± 4.5 mu;g/mL. Of the bacterial isolates, 1 colony with the VanC-1 genotype was identified as E. gallinarum by 16S rDNA sequencing, whereas the other 63 colonies had the VanC-2/3 genotype and were identified as E. casseliflavus. Although these results imply that the major head bays of Korea are not contaminated with the highly vancomycin-resistant VanA- or VanB-type VREs, the misuse of antibiotics should be prohibited to minimize the presence of VREs and to maintain a safe water supply for protecting the health of the general population. Based on the study results, we also recommend the implementation of a continuous, broad-spectrum inspection program for Enterococcus spp. and VRE contamination in the major head bays. Furthermore, the multiplex PCR method described in this study can be used effectively for this purpose. © 2012 Elsevier GmbH.

Lee G.-C.,Water Analysis and Research Center | Jeon E.-S.,Chungbuk National University | Kim W.-S.,Chungbuk National University | Le D.T.,Research Team for Vectorborne Diseases | And 2 more authors.
Virology Journal | Year: 2010

Background. This study evaluated the clinical accuracy and analytical sensitivity of the NanoSign® Influenza A/B antigen kit in detecting 2009 pandemic influenza A/H1N1 viruses. The kit is one of the most popular rapid diagnostic tests for detecting influenza in Republic of Korea. Results. The NanoSign® Influenza A/B kit resulted in 79.4% sensitivity and 97.2% specificity compared to RT-PCR in the detection of the viruses from 1,023 specimens. In addition, the kit was able to detect two strains of novel influenza viruses, Influenza A/California/12/2009(H1N1) and clinically isolated wild-type novel influenza A/H1N1, both of which are spreading epidemically throughout the world. In addition, the correlation between NanoSign® Influenza A/B test and conventional RT-PCR was approximately 94%, indicating a high concordance rate. Analytical sensitivity of the kit was approximately 73 3.65 ng/mL of the purified viral proteins and 1.13 0.11 hemagglutination units for the cultured virus. Conclusions. As the NanoSign® Influenza A/B kit showed relatively high sensitivity and specificity and the good correlation with RT-PCR, it will be very useful in the early control of influenza infection and in helping physicians in making early treatment decisions. © 2010 Lee et al; licensee BioMed Central Ltd.

Lee G.-C.,Water Analysis and Research Center | Jung G.S.,Chungbuk National University | Lee C.H.,Chungbuk National University
Virus Genes | Year: 2012

The complete nucleotide and deduced amino acid sequences of the RNA genome of a recently isolated norovirus (NoV) from Korea, designated Hu/GII-4/CBNU2/ 2007/KR (CBNU2), were determined and characterized by phylogenetic comparison with several genetically diverse NoV sequences. The RNA genome of CBNU2 is 7,560 nucleotides in length, excluding the 30 poly (A) tract. It includes three open reading frames (ORFs): ORF1, which encodes the nonstructural polyprotein (5-5,104); ORF2, which encodes VP1 (5,085-6,707); and ORF3, which encodes VP2 (6,707-7,513). ORF2-based phylogenetic analysis revealed that CBNU2 belonged to the GII.4 genotype, the most prevalent genotype, and formed a cluster with NoVs isolated from Asian regions, between 2006 and 2008. Comparative analysis with the consensus sequence of 207 completely sequenced NoV genomes showed 47 mismatched nucleotides: 26 in ORF1, 14 in ORF2, and 7 in ORF3, resulting in 8 amino acid changes: 3 in ORF1, 2 in ORF2, and 3 in ORF3. Phylogenetic analysis with full genome ORF1, ORF2, and ORF3 nucleotide sequences obtained from CBNU2 and each of the other representative NoV genomes suggested that CBNU2 had not undergone recombination with any of the other NoVs. A SimPlot analysis further supported this finding. © Springer Science+Business Media, LLC 2012.

Lee G.-C.,Water Analysis and Research Center | Kim M.-J.,Water Analysis and Research Center | Nam S.,Water Analysis and Research Center | Lee C.H.,Chungbuk National University
Microbiology and Immunology | Year: 2014

Several bodies of surface water in Korea were surveyed for the presence of hepatitis A virus (HAV) between 2007 and 2010. Of 265 surface water samples, 9 (3.4%) were HAV-positive. HAVs were mainly detected in the summer (3/62, 4.8%) and spring (4/96, 4.2%) seasons. Comparing different water sources, the highest prevalence (6.6%) of positive samples was seen in lake water, four HAV-positive samples being from lakes. Comparing prevalence rates across the four representative Korean basin systems, no HAVs were found in the Han or Nakdong river basins. The highest HAV prevalence was found in samples from the Yeongsan river and other basins (6.3%); the Geum/Seom river was also found to have a high HAV prevalence (5.7%). HAVs from the nine positive samples were then sequenced and analyzed phylogenetically. Two of the HAVs belong to genotype IA and fall within the same cluster as HAVs 6-3(ASAN4) (EU049548), KANSAN-PS1 (EU049554), and ASAN-KM (EU049563), which were collected from the stools of patients with gastroenteritis in Korea. The seven other HAV nucleotide sequences belong to the genotype IB cluster. This is the first nationwide surveillance of HAV in major Korean water sources. © 2014 The Societies and Wiley Publishing Asia Pty Ltd.

Lee J.H.,Catholic Kwandong University | Lee G.-C.,Water Analysis and Research Center | Kim J.I.,Chungbuk National University | Yi H.A.,Chungbuk National University | Lee C.H.,Chungbuk National University
Journal of Virological Methods | Year: 2013

The development of rapid and effective methods to detect water- and food-borne enteric viruses is important for the prevention and control of mass infection. This study represents an attempt to develop a reliable cell culture-based detection system and optimize an effective and rapid protocol for the assaying of environmental samples for the presence of infectious enteric viruses. Six enteric viruses were used in this study: poliovirus, Coxsackie virus A9, Coxsackie virus B5, human rotavirus G1, hepatitis A virus, and adenovirus type 41. Among the cell lines from humans (A549, HeLa, HEK293, and HFF) and other primates (Vero, BS-C-1, FRhK-4, BGMK, and MA104), a cytopathic effect (CPE) analysis indicated that the MA104 cell line was the most optimal for use in the detection of infectious enteric viruses. Both the sensitivity and specificity of virus detection in MA104 cells were similar to or higher than those in standard BGMK cells. Next, a method was developed for the determination of the infectiousness of enteric viruses using the colorimetric thiazolyl blue (MTT) assay. This assay utilizes 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide to yield % values based on colorimetric results. These results were compared with those from a conventional CPE-based TCID50 assay, revealing no statistically significant difference between the two methods. The MTT% values in MA104 cells were comparable to those in BGMK cells. This MA104 cell-based MTT assay could substitute for the classical BGMK cell-based CPE assay for infectious enteric viruses. © 2013 Elsevier B.V.

Lee G.-C.,Water Analysis and Research Center | Lee J.H.,Catholic Kwandong University | Kim B.Y.,Chungbuk National University | Lee C.H.,Chungbuk National University
Journal of Microbiology and Biotechnology | Year: 2013

Mitochondria often play central roles in apoptotic pathways, and disruption of the mitochondrial transmembrane potential (ΔΨm) has been observed in various cells undergoing apoptosis. Human cytomegalovirus (HCMV) infection induces apoptosis in permissive cells; however, investigations of mitochondria-targeted apoptosis in HCMV-infected human foreskin fibroblast (HFF) cells have been limited. Here, we investigated the mitochondrial apoptosis pathway in HCMV-infected HFF cells. Flow cytometry analysis using JC-1 revealed that HCMV infection induces disruption of ΔΨm in HFF cells when administered 24 h postinfection (hpi), and this disruption was maximized at 48 hpi. Moreover, cytochrome c, normally a mitochondrial inner membrane protein, was detected in cytoplasmic extracts of HCMV-infected cells, but not mock-infected cells, by western blot analysis at 24 hpi. A caspase activity assay based on fluorescence spectrophotometry using a fluorogenic substrate revealed an increase in caspase-3 activity at 48 hpi in HCMV-infected cells. Caspase-8 activity was increased at 72 hpi in HCMV-infected cells. These results imply that HCMV infection induces mitochondria-mediated apoptosis in HFF cells. © 2013 by The Korean Society for Microbiology and Biotechnology.

He W.,Sejong University | Choi I.,Water Analysis and Research Center | Lee J.-J.,Daegu University | Hur J.,Sejong University
Science of the Total Environment | Year: 2016

In this study, temporal and spatial variations in five defined molecular size fractions of dissolved organic matter (DOM) were examined for a well preserved wetland (Upo Wetland) and its surrounding areas, and the influencing factors were explored with many biotic and abioic parameters. For each DOM sample, the five size fractions were determined by size-exclusion chromatography coupled with organic carbon detector (SEC-OCD). For 2-year long monthly monitoring, bio-polymers (BP), humic substances (HS), building blocks (BB), low molecular-weight (LMW) neutrals, and LMW acids displayed the median values of 264, 1884, 1070, 1090, and 11 μg-C L-1, respectively, accounting for 6.2%, 41.7%, 24.5%, 26.4%, and 0.4% of dissolved organic carbon (DOC). The dominant presence of HS indicated that terrestrial input played important roles in DOM composition of the freshwater ecosystem, which contrasted with coastal wetlands in other reports. Both seasonal and periodic patterns in the variations were found only for HS and BB among the size fractions. It was also notable that the sources of HS were seasonally shifted from aquagenic origin in winter to pedogenic origin in summer. The correlations among the size fractions revealed that BB and LMW neutrals might be degradation products from HS and humic-like substances (HS + BB), respectively, while LMW acids, from LMW neutrals. Principle component analysis revealed that the humic-like substances and the aromaticity of DOM were associated with temperature, chlorophyll a, phosphorous, and rainfall, whereas the other fractions and the molecular weight of HS were primarily affected by solar irradiation. Significant correlations between DOM composition and some biotic factors further suggested that DOM may even affect the biological communities, which provides an insight into the potential coupling effects of biotic and abiotic factors on DOM molecular composition in freshwater wetlands. © 2015 Elsevier B.V.

Quang V.L.,Sejong University | Choi I.,Water Analysis and Research Center | Hur J.,Sejong University
Environmental Science and Pollution Research | Year: 2015

In this study, five different dissolved organic matter (DOM) fractions, defined based on a size exclusion chromatography with simultaneous detection of organic carbon (OCD) and ultraviolet (UVD), were quantitatively tracked with a treatment train of coagulation/flocculation–sand filtration–ozonation–granular activated carbon (GAC) filtration in a full-scale advanced drinking water treatment plant (DWTP). Five DOM samples including raw water were taken after each treatment process in the DWTP every month over the period of three years. A higher abundance of biopolymer (BP) fraction was found in the raw water during spring and winter than in the other seasons, suggesting an influence of algal bloom and/or meltwater on DOM composition. The greater extent of removal was observed upon the coagulation/flocculation for high-molecular-weight fractions including BP and humic substances (HS) and aromatic moieties, while lower sized fractions were preferentially removed by the GAC filtration. Ozone treatment produced the fraction of low-molecular-weight neutrals probably resulting from the breakdown of double-bonded carbon structures by ozone oxidation. Coagulation/flocculation was the only process that revealed significant effects of influent DOM composition on the treatment efficiency, as revealed by a high correlation between the DOM removal rate and the relative abundance of HS for the raw water. Our study demonstrated that SEC-OCD-UVD was successful in monitoring size-based DOM composition for the advanced DWTP, providing an insight into optimizing the treatment options and the operational conditions for the removal of particular fractions within the bulk DOM. © 2015, Springer-Verlag Berlin Heidelberg.

PubMed | Water Analysis and Research Center and Sejong University
Type: Journal Article | Journal: Environmental science and pollution research international | Year: 2016

Understanding the roles of inland reservoirs becomes increasingly important with respect to global carbon cycling as well as water resource management due to the unprecedented demand for construction in recent decades. In this study, the dissolved organic matter (DOM) quantity and quality in a newly constructed dam reservoir and its tributaries were monitored monthly during the initial impounding period (July to November 2014) using a size exclusion chromatography (SEC) with online organic carbon detector (OCD). The highest values were observed in the month of August with the highest precipitation for the bulk dissolved organic carbon (DOC), specific UV absorbance (SUVA), and most of the assigned size fractions (except for biopolymers) in the tributaries, indicating that allochthonous sources of DOM were dominant in the feeding stream waters of the reservoir. The bulk DOC and high molecular weight humic substance fraction (1kDa) were generally co-varied with the monthly precipitation in the tributaries, while building blocks (350-500Da), and low molecular weight (LMW) acids and neutrals showed different trends. In a dam site, the smaller molecular fractions became more abundant during the dry season (September to November), presumably due to the in-reservoir processes such as photo- and bio-degradation. Our results also revealed that storms mobilized a large amount of highly aromatic soil-derived DOM to the reservoir. A depth profile at the dam site showed the water is well mixed up to a depth of 20m. The SEC-OCD data coupled with non-metric multidimensional scaling provided a clear visualization of the spatiotemporal variations in DOM composition, which shed new light on the DOM composition formed in a newly constructed dam reservoir and also on the strategies for future water treatment options.

Nam S.,Water Analysis and Research Center | Lee G.,Water Analysis and Research Center
Biomedical and Environmental Sciences | Year: 2010

Objective To simultaneously detect viable Cryptosporidium parvum oocysts and Giardia duodenalis cysts for the purpose of reducing time and cost spent. Methods A duplex reverse transcription polymerase chain reaction (RT-PCR) method was newly developed. Results Using duplex RT-PCR method for the hsp70 gene, viable (oo)cyst concentrations of 101 and 103 (oo)cysts/100 μL could be detected for C. parvum and G duodenalis, respectively. However, after heat-shock stimulation the expression oihsp70 mRNAs was detectable at 10° an10(oo)cysts/100 μL concentrations of C. parvum and G duodenalis, respectively. Thus, the detection sensitivity was significantly increased when the viable (oo)cysts were exposed to heat shock. Conclusion This study describes a new duplex RT-PCR method for hsp70 gene to detect the viable (oo)cysts of the C. parvum and G duodenalis with less time consumed and at a lower cost. This newly developed duplex RT-PCR method may be used to detect these parasites not only in aquatic environments but also in clinical samples. Copyright © 2010 by China CDC.

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