-
[show abstract]
[hide abstract]
ABSTRACT: A highly significant increase in anti-Vesivirus (family Caliciviridae) antibody prevalence, along the axis from healthy blood donors; donors with elevated transaminase; patients with clinical hepatitis; and patients with post-transfusion/dialysis hepatitis, has been reported in human sera from the USA and Europe. Asian samples have now been tested retrospectively using serology and enzyme immunoassay (EIA) with a Vesivirus partial-capsid antigen expressed as a fusion protein. Anti-vesiviral antibodies were measured by optical densities (OD(650) ) and compared in patients separated by age, gender and Groups A-F as follows: Control Group A, an Experimental Group B, which was divided further into Group C, patients with elevated enzymes (alanine transaminase (ALT), aspartate transaminase (AST), and γ-glutamyl transpeptidase (γ-GT); Group D, patients receiving transfused blood; Group E, patients with high enzyme levels after transfusion; and Group F, hepatitis B and C positive patients. Using multivariate logistic regression analyses, a significantly greater proportion of patients receiving transfusion(s), were positive for anti-Vesivirus antibody compared with non-transfused patients (P = 0.008; OR: 3.86, 95% CI: 1.43-10.43). Also, anti-Vesivirus antibody was significantly associated with elevated biochemical liver function tests: ALT ≥120 IU or AST ≥ 120 IU (P = 0.017; OR: 4.23, 95% CI: 1.30-13.80). In the blood transfusion group, anti-Vesivirus antibody was significantly correlated with high enzyme levels (ALT, P = 0.018; AST, P = 0.010; γ-GT, P = 0.020). These data provide serologic evidence of vesiviral transfusion-transmission-associated disease, which could include infection of any organ system where cytopathology resulted in high levels of either ALT or AST. J. Med. Virol. 84:1943-1952, 2012. © 2012 Wiley Periodicals, Inc.
Journal of Medical Virology 12/2012; 84(12):1943-52. · 2.82 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Norovirus (NoV) and hepatitis A virus (HAV) are pathogenic enteric viruses responsible for public health concerns worldwide. The viral transmission occurs through fecally contaminated food, water, fomites, or direct contact. However, the difficulty in cultivating these viruses makes it challenge to characterize the resistance to various environmental stresses. In this study, we characterized the inactivation rates of murine norovirus (MNV), MS2, and HAV on either lacquer coating rubber tree wood or stainless steel under different temperature and relative humidity (RH) conditions. The viruses were analyzed at temperatures of 15˚C, 25˚C, 32˚C, and 40˚C and at RHs of 30%, 50%, and 70% for 30 days. Overall, they survived significantly longer on wood than on steel at lower temperature (P < 0.05). The inactivation rate of MS2 and MNV increased at higher RH levels, whereas HAV survived the best at a medium RH level (50%). The effect of RH was significant only for MS2 (P < 0.05). MS2 persisted longest under all the environmental conditions examined. Both a linear and a nonlinear Weibull model were used to describe the viral inactivation data in this study. The data produced a better fit to the survival curves that were predicted by the Weibull model.
Environmental Science & Technology 11/2012; · 4.80 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Various enteric viruses including norovirus, rotavirus, adenovirus, and astrovirus are the major etiological agents of food-borne and water-borne disease outbreaks and frequently cause non-bacterial gastroenteritis worldwide. Sensitive and high-throughput detection methods for these viral pathogens are compulsory for diagnosing viral pathogens and subsequently improving public health. Hence, we developed a sensitive, specific, and high-throughput analytical assay to detect most major enteric viral pathogens using "Combimatrix" platform oligonucleotide probes. In order to detect four different enteric viral pathogens in a sensitive and simultaneous manner, we first developed a multiplex RT-PCR assay targeting partial gene sequences of these viruses with fluorescent labeling for the subsequent microarray. Then, five olignonucleotides specific to each of the four major enteric viruses were selected for the microarray from the oligonulceotide pools targeting the specific genes obtained by multiplex PCR of these viruses. The oligonucleotide microarray was evaluated against stool specimens containing single or mixed viral species. As a result, we demonstrated that the multiplex RT-PCR assay specifically amplified partial sequences of four enteric viruses and the subsequent microarray assay was capable of sensitive and simultaneous detection of those viruses. The developed method could be useful for diagnosing enteric viruses in both clinical and environmental specimens.
The Journal of Microbiology 10/2012; · 1.10 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Exposure to bioaerosols causes various adverse health effects including infectious and respiratory diseases, and hypersensitivity. Controlling exposure to bioaerosols is important for disease control and prevention. In this study, we evaluated the efficacies of various functional filters coated with antimicrobial chemicals in deactivating representative microorganisms on filters or as bioaerosols. Tested functional filters were coated with different chemicals that included (i) Ginkgo and sumac, (ii) Ag-apatite and guanidine phosphate, (iii) SiO2, ZnO, and Al2O3, and (iv) zeolite. To evaluate the filters, we used a model ventilation system (1) to evaluate the removal efficiency of bacteria (Escherichia coli and Legionella pneumophila), bacterial spores (Bacillus subtilis spore), and viruses (MS2 bacteriophage) on various functional filters, and (2) to characterize the removal efficiency of these bioaerosols. All experiments were performed at a constant temperature of 25 degrees C and humidity of 50%. Most bacteria (excluding B. subtilis) rapidly decreased on the functional filter. Therefore, we confirmed that functional filters have antimicrobial effects. Additionally, we evaluated the removal efficiency of various bioaerosols by these filters. We used a six-jet collision nebulizer to generate microbial aerosols and introduced it into the environmental chamber. We then measured the removal efficiency of functional filters with and without a medium-efficiency filter. Most bioaerosol concentrations did not significantly decrease by the functional filter only but decreased by a combination of functional and medium-efficiency filter. In conclusion, functional filters could facilitate biological removal of various bioaerosols, but physical removal of these by functional was minimal. Proper use of chemical-coated filter materials could reduce exposure to these agents.
Journal of Microbiology and Biotechnology 09/2012; 22(9):1288-95. · 1.38 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The ability to distinguish between viable and/or infectious micro-organisms and inactivated cells is extremely important for correctly performing microbial risk assessments. In this study, we evaluated whether propidium monoazide (PMA)-qPCR could distinguish between viable and nonviable bacteria and viruses.
A PMA-qPCR combined assay was applied to viable and inactivated bacteria (Escherichia coli and Bacillus subtilis) and viruses (MS2 and murine norovirus [MNV]). PMA, a DNA-intercalating agent, in combination with PCR was better able to distinguish between viable and nonviable bacteria and viruses than conventional PCR.
These results suggest that a combined PMA-qPCR assay can be used to measure the viability of bacterial cells and bacteriophage MS2, but not MNV.
PMA-qPCR could potentially be used to measure the viability of some micro-organisms, including virus. However, a thorough evaluation should be performed prior to measuring the viability of micro-organisms by PMA-qPCR in a quantitative way.
Letters in Applied Microbiology 06/2012; 55(3):182-8. · 1.62 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: We investigated the resistance of murine norovirus (MNV) and coliphage MS2, a culturable human norovirus surrogate, to temperature, salt, and pH. Virus inactivation was measured by plaque, real-time TaqMan reverse transcription (RT) PCR, and long-template RT-PCR assays. Both MNV and MS2 were rapidly inactivated at temperatures above 60°C. Similarly, MNV tolerated low salt concentrations (0.3% NaCl) to a greater degree than high salt concentrations (3.3 to 6.3% NaCl). MNV was relatively resistant to strong acidic conditions (pH 2) and was more tolerant of slightly acidic (pH 4) or neutral (pH 7) conditions. In contrast, MS2 was resistant to high salinity. Overall, temperature had a greater effect on infectivity than salt or low pH. Additionally, temperature and low pH had a synergistic effect on MNV infectivity. Both real-time and long-template RT-PCR assays significantly underestimated the inactivation by temperature, salt, and pH. The inactivation kinetics of both MNV and MS2 under various environmental conditions gave a good fit by the Weibull model (R² > 0.9). This study suggests both the capacity of infectious human norovirus to persist in the face of various environmental conditions and its sensitivity to high temperatures, which may provide a mechanism of protection against this virus.
Journal of food protection 03/2012; 75(3):533-40. · 1.94 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: We previously demonstrated that genotyping followed by proper statistical analyses of F plus (F+)-specific RNA coliphages can effectively represent fecal origins of either humans or animals. Here, we performed microbial source tracking (MST) using F+ RNA coliphages as a target MST microorganism for identifying fecal sources contaminating ground and surface water in metropolitan Seoul and Gyeonggi Province in South Korea. In total, 71 groundwater and 5 surface water samples were collected and screened for the presence of F+ RNA coliphages. More than 124 F+ coliphages were isolated from six groundwater and five surface water samples by the single agar layer method. F+ RNA coliphages were predominant in both waters (100% and 91%, respectively). Genotyping of 118 F+ RNA coliphages revealed that most (51/60) of the groundwater F+ RNA coliphages belonged to group I, whereas both groups I (25/58) and IV (31/58) were predominantly observed in surface water. Further comparison of phage isolates from human and animal (pig, cow, goose, and chicken) fecal sources using nucleic acid sequencing and principal coordinate analysis showed that groundwater samples formed clusters associated with cow feces, whereas surface waters formed clusters related to chicken and human feces. These results indicate the potential of the F+ RNA coliphage-based MST for identifying fecal contamination sources, which may be further exploited and validated in different geographical regions of the world.
Science of The Total Environment 12/2011; 412-413:127-31. · 3.29 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Seasonal and spatial variation in the genotypic richness of 3480 Escherichia coli isolates obtained from the Yeongsan River basin in South Korea was investigated by using the horizontal fluorophore-enhanced rep-PCR (HFERP) DNA fingerprinting technique. The relationship between 60 E. coli isolates from each of 58 freshwater samples was determined by using multidimensional scaling (MDS) analysis and self-organized maps (SOMs). The MDS analysis, done based on HFERP DNA fingerprints, showed that E. coli isolates obtained in October through December clustered tightly, while those obtained in other sampling periods were more genetically diverse. However, site-specific E. coli genotypes were not observed. SOMs analysis, done using the 10 most frequently isolated E. coli genotypes, showed the occurrence of season-specific E. coli genotypes and the main SOMs clusters were most influenced by temperature, strain diversity and biochemical oxygen demand. Diversity among E. coli genotypes tended to decrease as water temperature decreased, and the numbers of E. coli genotypes observed in urban area were greater, more diverse and less dependent on water temperature than those obtained from agricultural areas. Taken together, our findings indicate that that an ecological approach needs to be considered in order to obtain a better understanding of E. coli community dynamics in the environment and that SOMs analysis is useful to visualize the multidimensional dependent variables that are influencing the types and dynamics of specific E. coli genotypes in the environment.
Environmental Microbiology 08/2011; 13(12):3103-13. · 5.84 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: We compared the composition of the fecal microbiotas of Korean and U.S. adult twins. Our data indicated that the gut microbiota shows some signature of biogeography, potentially mediated by differences in diet and/or other environmental factors; however, these regional differences may be masked by other phenotypic variations, such as obesity.
Applied and environmental microbiology 08/2011; 77(20):7433-7. · 3.69 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Lactobacillus ruminis SPM0211 is a potential probiotic strain that shows antimicrobial activity against emerging pathogens. Here we present the draft genomic sequence of L. ruminis SPM0211, isolated from a fecal sample from a healthy Korean and describe both the common and unique features of this strain.
Journal of bacteriology 07/2011; 193(18):5034. · 3.94 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Norovirus (NoV) is the major etiological agent causing foodborne and waterborne outbreaks worldwide. We developed a novel duplex real-time quantitative RT-PCR assay designed for the simultaneous detection of and discrimination between NoV genogroups GI and GII, by targeting the short junction region between ORF1 and ORF2, with sensitivity and efficiency comparable to those of each simplex RT-PCR assay. This new duplex assay was evaluated against clinical stool (n = 82) and environmental (groundwater or surface water, n = 60) specimens from South Korea, and the results were compared with those of conventional RT-PCR (cRT-PCR) assays. The duplex assay detected more positive samples than did the cRT-PCR for both clinical (74 vs. 71) and, more strikingly, environmental (24 vs. 10) specimens. No cross-reactivity against specimens containing other enteric viruses such as rotavirus, adenovirus, and poliovirus were observed. These results suggest that this newly developed duplex real-time RT-PCR assay can be used for the sensitive and simultaneous genogroup-specific detection of NoV in both clinical and environmental specimens.
Virus Genes 06/2011; 43(2):192-200. · 1.85 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: South Korea uses more antibiotics for animal production than any other member of the OECD. The excessive use could potentially lead to a greater incidence of antibiotic resistant microorganisms, some of which may be pathogenic. In this study, we examined potential diarrheagenic and extraintestinal pathogenic E. coli (ExPEC) isolates using virulence gene profiling (eaeA, est, elt, ipaH, stx1 and stx2 as diarrheagenic and afa/dra, iutA, kpsMT II, papA/C, and sfa/foc as extraintestinal pathogenic virulence genes). A cluster analysis of DNA fingerprint data indicated that certain genotypes of chicken E. coli isolates are likely ExPEC. Moreover, our results showed 38 of the 60 potential diarrheagenic and ExPEC isolates obtained from animals and humans were also resistant to antibiotics, mostly to tetracycline groups, penicillin groups, and folate pathway inhibitors. Our results suggest that the use of antibiotics in agriculture should be controlled due to potential impacts on human health.
Microbes and Environments 05/2011; 26(3):198-204. · 1.91 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Vitamin A supplementation is associated with divergent clinical norovirus (NoV) outcomes in Mexican children. Fecal cytokine concentrations following NoV genogroup infections among 127 Mexican children 5-15 mo old enrolled in a randomized, double-blind, placebo-controlled, vitamin A supplementation trial were determined to clarify the role the gut immune response plays in these associations. Stools collected from supplemented children [20,000 IU retinol (3.3 IU = 1 μg retinol) for children < 12 mo of age; 45,000 iu for children ≥ 12 mo] or children in the placebo group were screened for NoV genogroups I (GI) and II (GII). Monocyte chemoattractant protein-1 (MCP-1), TNFα, IL-5, IL-6, IL-8, IL-4, IFNγ, and IL-10 fecal concentrations were also determined. Differences in cytokine levels between the 2 groups following GI and GII infections were determined using ordered logistic regression models. MCP-1 and IL-8 levels were greater among GI- and GII-infected children, respectively, compared with uninfected children, whereas IL-5 levels were greater following both genogroup infections. MCP-1, IL-8, and IL-6 fecal levels were reduced among supplemented children with GII-associated diarrhea compared with the placebo group. Vitamin A-supplemented, GII-infected children had reduced MCP-1 and TNFα levels compared with GII-infected children in the placebo group (P-interaction = 0.02 and 0.03, respectively). Supplemented children with GI-associated diarrhea had higher TNFα and IL-4 levels compared with children in the placebo group with diarrhea (P-interaction = 0.02 and 0.02, respectively). The divergent effects of supplementation on NoV outcomes may result from the different effects vitamin A has on the genogroup-specific immune responses.
Journal of Nutrition 03/2011; 141(5):957-63. · 3.92 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Groundwater is an important source of drinking and household water worldwide. Hence, the quality of groundwater is very important for preventing waterborne disease outbreaks and should be properly monitored. This study investigated the prevalence of waterborne viruses and fecal indicators in groundwater in metropolitan Seoul and Gyeonggi province, South Korea. A total of 116 samples of groundwater were taken using NanoCeram filters during both summer (June to August) and fall-winter seasons (October to December) in 2008. Among 71 sampling sites, 28 (48.3%) and 18 (35.3%) were positive for norovirus (NoV) from the summer and fall-winter season, respectively. The identified genotypes of NoV include GI-1, 4, 8, 9 and GII-4, 10, 11 (or 17), 13, 15 (or 16). None of fecal indicators was significantly correlated with NoV in groundwater. Among the tested fecal indicators, somatic coliphage (95.3%) showed an excellent true-negative rate of NoV occurrence. The combination of chemical, microbial and viral indicators increased the positive predictive value (50-100%). This study demonstrated a high prevalence of NoV in groundwater in metropolitan Seoul areas and characterized the positive and negative predictive values of a fecal indicator for predicting NoV prevalence.
Science of The Total Environment 03/2011; 409(11):2078-84. · 3.29 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The virus adsorption-elution (VIRADEL) technique has been widely used in the recovery of various enteric viruses in water, and an electropositive filter such as 1 MDS has been commonly applied. However, effective methods of monitoring waterborne norovirus (NoV) have not yet been well characterized and optimized. Hence, in this study, the VIRADEL technique was evaluated and optimized for effectively detecting NoV in water by two commonly used electropositive filters (1MDS and NanoCeram filter). Various elution and concentration methods were evaluated by using both murine norovirus (MNV) and human NoV. Among the tested elution buffers, the most effective was 1.5% beef extract plus 0.01% Tween 80 for both 1MDS (67.5%) and NanoCeram (85.7%) microfilters. The recovery rate of GII-4 human NoV was higher by organic flocculation (86.6%) than by polyethylene glycol (PEG) precipitations (11.6~73.6%). When both 1MDS and NanoCeram filters were tested to detect NoV in surface and groundwater, the sensitivity of NoV recovered by these filters appeared to depend on the types and conditions of environmental water. The results of this study will help to set a standard of detection method for NoV in water.
Journal of Water and Health 03/2011; 9(1):27-36. · 1.37 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Microbial compositions of human and animal feces from South Korea were analyzed and characterized. In total, 38 fecal samples (14 healthy adult humans, 6 chickens, 6 cows, 6 pigs and 6 geese) were analyzed by 454 pyrosequencing of the V2 region of the 16S rRNA gene. Four major phyla, Actinobacteria, Proteobacteria, Firmicutes and Bacteroidetes, were identified in the samples. Principal coordinate analysis suggested that microbiota from the same host species generally clustered, with the exception of those from humans, which exhibited sample-specific compositions. A network-based analysis revealed that several operational taxonomic units (OTUs), such as Lactobacillus sp., Clostridium sp. and Prevotella sp., were commonly identified in all fecal sources. Other OTUs were present only in fecal samples from a single organism. For example, Yania sp. and Bifidobacterium sp. were identified specifically in chicken and human fecal samples, respectively. These specific OTUs or their respective biological markers could be useful for identifying the sources of fecal contamination in water by microbial source tracking.
The ISME Journal 02/2011; 5(2):362-5. · 7.38 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: To inspect the norovirus contamination of groundwater in South Korea, a nationwide study was performed in the summer (June to August) and winter (October to December) of 2008. Three-hundred sites designated by the government ministry were inspected. Water samples were collected for analysis of water quality, microorganism content, and viral content. Water quality was assessed by temperature, pH, turbidity, residual chlorine, and nitrite nitrogen content. Microorganism contents were analyzed bacteria, total coliforms, Escherichia coli, and bacteriophage. Virus analyses included panenterovirus and norovirus. Two primer sets were used for the detection of norovirus genotypes GI and GII, respectively. Of 300 samples, 65 (21.7%) were norovirus positive in the summer and in 52 (17.3%) were norovirus positive in the winter. The genogroup GI noroviruses that were identified were GI-1, GI-2, GI-3, GI-4, GI-5, GI-6, and GI-8 genotypes; those in the GII genogroup were GII-4 and GII-Yuri genotypes. The analytic data showed correlative relationships between the norovirus detection rate and the following parameters: water temperature and turbidity in physical-chemical parameters and somatic phage in microbial parameters. It is necessary to periodically monitor waterborne viruses that frequently cause epidemic food poisoning in South Korea for better public health and sanitary conditions.
Applied and environmental microbiology 02/2011; 77(4):1466-74. · 3.69 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: The microbial content of indoor air is an increasingly important issue. High microbial load is associated with various adverse health effects, such as infectious disease, asthma, and toxicosis. The most commonly applied method for analyzing airborne microorganisms is a six-stage Anderson microbiological impactor and subsequent cultivation on agar plates. However, such culture can take several days to produce results, and only a small proportion of viable microorganisms are culturable. Thus, cultivation cannot rapidly identify the degree of health risk or fully characterize microbial communities related to adverse health effects. We sampled bioaerosols using both a high volume SpinCon sampler and a six-stage Andersen microbial impactor. These samples were assayed by culturing, real-time TaqMan PCR, and ATP bioluminescence. We found a significant positive association between real-time TaqMan and cultivation, but not between total ATP and cultivation. Thus, a real-time TaqMan PCR assay could be used in place of cultivation to assess microbial air quality. Finally, when microbial communities were characterized using denaturing gradient gel electrophoresis (DGGE) and principal component analysis (PCA), a seasonal clustering of microbial profiles was observed.
Science of The Total Environment 02/2011; 409(9):1732-7. · 3.29 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: Enterococcus isolates (1500) obtained from the feces of 48 humans, 209 domesticated food animals, and 155 wild geese in South Korea were characterized with respect to species status by PCR analyses and resistance to antibiotics. Of the 1500 strains examined, the majority (n=577) were Enterococcus faecalis from 224 (54.4%) of the samples feces, while 299 were of E. faecium from 125 of the samples (30.3%), 224 were E. hirae from 101 (24.5%) of the samples, 94 were E. casseliflavus from 43 (10.4%) of the samples, and one was E. gallinarum. While 305 isolated from 125 (30.3%) of the samples were unidentified species. Approximately 15, 60, 50, 55, 3, and 40% of samples obtained from beef cattle, chickens, ducks, swine, wild geese, and humans, respectively, yielded Enterococcus isolates that were resistant to high-levels of aminoglycosides (i.e., of gentamicin, kanamycin, and streptomycin, minimum inhibitory concentrations were >1000 mg/l). The 180 Enterococcus isolates that showed high levels of resistance to aminoglycoside antibiotics (HLAR) were screened for virulence genes encoding for aggregation substance (agg), cytolysin activator (cylA), gelatinase (gelE) and surface protein (esp). Of those, the gelE gene was found most frequently in chickens and ducks of the HLAR isolates, while 56 E. faecalis and 13 E. faecium HLAR were gelatinase positive and showed hemolysin activity. Multiple antibiotic resistant Enterococcus isolates carrying virulence genes were most frequently isolated from poultry and swine, and were mostly E. faecalis or E. faecium. These findings suggest that restriction of the use of antibiotics in food animal operations in South Korea, especially those involved in poultry and swine production would be desirable.
International journal of food microbiology 10/2010; 144(3):387-92. · 3.01 Impact Factor
-
[show abstract]
[hide abstract]
ABSTRACT: We evaluated and optimized each step in an analytical method for detecting norovirus from various foods. We characterized the buffers needed for eluting norovirus from foods such as ham and lettuce. Two different concentration methods, polyethylene glycol (PEG) precipitation and hollow fiber ultrafiltration (HUF), were compared using both murine norovirus (MNV) and human norovirus (HuNoV). For PEG precipitation, an elution buffer containing 3% beef extract (pH 7.1) was more suitable than 0.05 M glycine plus 0.14 M NaCl (pH 7.5), and the recovery efficiency increased with increasing molecular weight of PEG. To determine the optimal buffer for concentrating norovirus by HUF, glycine buffers with different pH values and ionic strengths were examined as elution buffers. Overall, HUF was more efficient for norovirus recovery than was PEG precipitation. Because there was a significant positive correlation between MNV and HuNoV results, MNV could be a useful surrogate for detecting HuNoV in foods.
Journal of food protection 09/2010; 73(9):1651-7. · 1.94 Impact Factor
