• Korean Journal of Microbiology
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• v.28 no.3
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• pp.229-235
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• 1990

A rapid and sensitive method to detect the extracellular enzymatic activity of bacteria colonies grown on agar plates is described. Selective agar media supplemented with protein, starch, chitin, Tween-80, etc. are conventionally used to detect biochemical properties of bacteria. It has been experimentally demonstrated with bacteria pure cultures that fluorogenic Methylumbelliferyl (MUF)-substrates are excellent substrate analogues for normally occurring polymers. Based on MUF-substrate hydrolysis the new method provides reliable qualitative estimates of extracellular enzymatic properties of bacteria within minutes using pure cultures as well as agar p;ates prepared for colony counts. Extracellular enzyme activities of heterotrophic bacteria from freshwater ecosystems and marine sediment using this method are discussed.

• Journal of Industrial Technology
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• v.21 no.B
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• pp.11-20
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• 2001

This laboratory study examined the impact of free chlorine residual and pipe material on the formation of biofilm in drinking water distribution pipe surfaces. Result of heterotrophic plate counts(HPC) of the biofilm in the tap water-supplied reactor averaged $2.17{\times}10^5CFU/cm^2$ on PVC and $2.43{\times}10^5CFU/cm^2$ on STS 316, respectively. HPCs on the surface exposed to the tap water containing 0.2mg/L of free chlorinne residual averaged $4.24{\times}10^4CFU/cm^2$ on PVC and $6.54{\times}10^4CFU/cm^2$ on STS 316, respectively. Average of HPC/Total direct counts in the tap water-supplied reactor ranged from 1.08%(PVC) to 1.26%(STS 316) and from 0.38%(PVC) to 0.65%(STS 316) in the reactor supplemented with disinfectant, respectively. No correlation was observed between disinfectant addition and biofilm density. With regard to the biofilm formation, little difference existed between PVC and STS 316. Yellow and red pigmented bacteria were the dominant expressions in bulk fluid, whereas non-pigmented bacteria were found dominant in the biofilm. Pink/red pigmented bacteria were found to be facultative anaerobic, while yellow pigmented bacteria and non-pigmented bacteria were found to be obligate aerobic.

• Journal of Microbiology and Biotechnology
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• v.27 no.5
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• pp.995-1004
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• 2017

Culture-dependent methods, such as heterotrophic plate counting (HPC), are usually applied to evaluate the bacteriological quality of hemodialysis water. However, these methods cannot detect the uncultured or viable but non-culturable (VBNC) bacteria, both of which may be quantitatively predominant throughout the hemodialysis water treatment system. Therefore, propidium monoazide (PMA)-qPCR associated with HPC was used together to profile the distribution of the total viable bacteria in such a system. Moreover, high-throughput sequencing of 16S rRNA gene amplicons was utilized to analyze the microbial community structure and diversity. The HPC results indicated that the total bacterial counts conformed to the standards, yet the bacteria amounts were abruptly enhanced after carbon filter treatment. Nevertheless, the bacterial counts detected by PMA-qPCR, with the highest levels of $2.14{\times}10^7copies/100ml$ in softener water, were much higher than the corresponding HPC results, which demonstrated the occurrence of numerous uncultured or VBNC bacteria among the entire system before reverse osmosis (RO). In addition, the microbial community structure was very different and the diversity was enhanced after the carbon filter. Although the diversity was minimized after RO treatment, pathogens such as Escherichia could still be detected in the RO effluent. In general, both the amounts of bacteria and the complexity of microbial community in the hemodialysis water treatment system revealed by molecular approaches were much higher than by traditional method. These results suggested the higher health risk potential for hemodialysis patients from the up-to-standard water. The treatment process could also be optimized, based on the results of this study.

• Korean Journal of Microbiology
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• v.49 no.4
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• pp.424-428
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• 2013

To identify risk factors for Legionella contamination, water quality variables routinely measured in examination of natural and city waters were meta-analyzed for significance of correlation to Legionella incidences. For evaluation of abundance of Escherichia coli as a risk factor, which is currently used as an indicator of Legionella contamination in an official guideline in Korea, odds ratio (OR) of above-cutoff total coliform counts for Legionella presence/absence was used as the effect size in the meta-analysis. The OR was estimated as 1.05 (0.36-3.12, 95% CI), and the probability of having identical odds reached 0.92. Also, ORs from individual studies showed significant heterogeneity (P=0.008), which contributed to 63% of total variance of the ORs. In the case of heterotrophic plate count (HPC), the OR for Legionella presence/absence was 2.72 (2.04-3.63) with highly significant deviation from identical odds (P<0.0001). ORs from different studies were seemingly homogeneous ($Q_{df=8}$=12.7, P=0.12). Turbidity and concentrations of chlorine, iron ion and cupper ion were other routine variables that could be considered as risk factors. However, statistical measures from different studies were not uniform enough to develop an appropriate effect size while the number of studies reporting the variables was also small (3-5). In conclusion, HPC appeared to be appropriate as indicator of Legionella contamination, rather than fecal bacteria contamination. HPC may imply abundance of habitats (amoebas and biofilms) of Legionella in water. This result warrants further studies for standardizing protocols and cutoff values to infer Legionella risks from HPC.

• Journal of Environmental Health Sciences
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• v.41 no.6
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• pp.397-404
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• 2015

Objectives: The aim of this study is to investigate microbial contamination in the school food service environment for the assessment of microbial food safety. Methods: We collected both swab samples from tables and desks and airborne bacterial samples from an elementary school (School A) and a high school (School B). Heterotrophic plate count, total coliform, Staphylococcus aureus, and Bacillus cereus were measured with selective media to quantify microbial concentration. PCR assay targeting 16S rRNA genes was performed to identify the strains of S. aureus and B. cereus isolated. In addition, we made a food service checklist for the locations to evaluate the food service environment. A Wilcoxon test was employed to examine the differences in microbial concentration between before lunchtime and afterwards. Results: Heterotrophic plate counts showed higher levels after-lunch compared to before-lunch at School B. However, levels of S. aureus were higher in the after-lunch period (p<0.05) in both classrooms and in the cafeteria in School A. B. cereus was only sparsely detected in School B. Several samples from food dining carts were found to be contaminated with bacteria, and facilities associated with food delivery were found to be vulnerable to bacterial contamination. Although microbial concentrations in the air showed little difference between before- and after-lunchtime in the cafeteria in School A, those in classrooms were greater after-lunchtime at both schools. Conclusion: Our results suggested that the microbial safety in schools after lunchtime of concern. Necessary preventive measures such as hygiene education for students and food handlers should be required to minimize microbial contamination during food service processes in schools.

• Korean Journal of Microbiology
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• v.44 no.3
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• pp.244-250
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• 2008

To evaluate bacteriological water quality, samples were taken from drinking water dispensers placed at S company (S-C) and U highschool (U-H) in Ulsan. The medians of heterotrophic plate counts (HPCs) were 53 CFU/ml for the 74 water samples of S-C and 80 CFU/ml for the 36 cold water samples of U-H, and 38% of the S-C and 42% of the U-H samples showed HPC bacterial concentrations higher than 100 CFU/ml. Coliform bacteria were detected from one sample of S-C. To determine the major source of bacterial contamination, water samples were taken daily for $6\sim8$ days from the bottled water containers as well as the faucets of an experimental water dispenser. While the average HPCs in the bottled water containers were 33 CFU/ml for the first and 132 CFU/ml for the 2nd analysis, the HPC concentration in the cold water samples was 1,022 CFU/ml for the 2nd analysis. These results suggest that the majority of bacteria detected in the cold water samples were originated from the biofilms on the surface of water passages within the water dispensers. There was no significant increase in HPC bacterial concentrations within the bottled water container after installation on the water dispenser. We could isolate and tentatively identify 3 genera 6 species of Gram-positive and 7 genera 7 species of Gram-negative bacteria from the plate count agar plates of U-H samples. Among the isolates, 72% were observed as Gram-positive, and Micrococcus spp. was the most abundant with 54% of the total, followed by Sphingomonas paucimobilis with 16%. It appears that most of the HPC bacteria detected in water dispensers originate from indoor airborne bacteria, which may play important roles in the formation of biofilms on the surface of water passages within the water dispensers.

• Journal of Korean Society on Water Environment
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• v.36 no.4
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• pp.275-283
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• 2020

Fecal coliforms are indicator bacteria to evaluate fecal contamination and microbiological safety in environment water. To examine fecal coliforms by membrane filtration, 1% rosolic acid solution dissolved in sodium hydroxide(0.2 M) should be added to m-FC medium according to Korean standard method. To reduce the exposure of researchers to harmful chemicals and expenditure of unnecessary cost, we evaluated if the rosolic acid solution is required to detect fecal coliforms. For 113 samples collected from five intake sources of Seoul, 42 samples of six tributaries, and 11 samples of sewage, the number of fecal coliforms was compared in medium with or without the reagent. As a result, the number was higher in m-FC medium without the reagent, but there was not a statistically significant difference. In the water intake, m-FC medium without the reagent could be used to examine fecal coliforms except in July, August and in case of rainfall. When heterotrophic plate counts exceeded 1,000 CFU/filter, or during rainfall, there was an effect of background bacteria in two types of the medium. However, it was more appropriate to use m-FC medium with the reagent to suppress gram-positive bacteria that can grow on medium without the reagent. In the tributary and sewage samples, the effect of the background bacteria was low, allowing the use of medium without the reagent regardless rainfall. Thus, it is necessary to present in standard method that the addition of rosolic acid solution in m-FC medium can be selected according to the characteristics of samples.

• Journal of Korean Society of Environmental Engineers
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• v.27 no.12
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• pp.1311-1320
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• 2005

The purpose of this research is to survey characteristics of microbial community and the removal efficiency of organic materials for biological activated carbon in water treatment plant. Coal based activated carbon retained more attached bacterial biomass on the surface of the activated carbon than the other activated carbon with operating time and materials. The heterotrophic plate count(HPC), eubacteria(EUB) and 4,6-diamidino-2-phenylindole(DAPI) counts were ranged from $0.95{\times}10^7$ to $52.4{\times}10^7$ CFU/g, from $3.8{\times}10^8$ to $134.2{\times}10^8$ cells/g and from $7.0{\times}10^8$ to $250.2{\times}10^8$ cells/g, respectively. The biomass of EUB and DAPI appeared to be much more $10^2$ than HPC, which were increasing in bed volume of 20,000 at the stage of steady-state. The change of microbial community by analyzing fluorescent in situ hybridization(FISH) method with rRNA-targeted oligonucleotide probes, the dominant group was $\alpha$-proteobacteria($\alpha$ group) and high G+C content bacteria(HGC) the lowest distributing rate before reaching the bed volume of 20,000. After reaching the bed volume of 20,000, $\alpha$ group and other groups of bacteria became decreased, on the other hand, the proportion of both $\beta$-proteobacteria($\beta$ group) and $\gamma$-proteobacteri($\gamma$ group) were increasing. Coconut and wood based activated carbons had similar trend with coal based activated carbon, but the rate of $\alpha$ group on coal based activated carbon had gradually increased. Bacterial production with the operating period appeared highest in coal based activated carbon at the range of $1.2{\sim}3.4\;mg-C/m^3{\cdot}h$ while the coconut and wood based activated carbon were ranged from 1.1 to 2.6 $mg-C/m^3{\cdot}h$ and from 0.7 to 3.5 $mg-C/m^3{\cdot}h$ respectively. The removal efficiency of assimilable organic carbon(AOC) showed to be highly correlated with bacterial production. The correlation coefficient between removal efficiency of AOC and bacterial production were 0.679 at wood based activated carbon, 0.291 at coconut based activated carbon and 0.762 at coal based activated carbon, respectively.

• Journal of Environmental Health Sciences
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• v.39 no.6
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• pp.513-521
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• 2013

Objectives: This study was performed in order to detect indicator bacteria in drinking spring water (DSW) samples in Seoul Metropolitan City, and to identify their genus through 16S rRNA sequencing and then assessing the genetic relation of their strains. Methods: For indicator bacteria detection and identification of total coliforms, we analyzed DSW between the spring and summer seasons. In particular, DSW samples were chosen from sites repeatedly found unsatisfactory in recent years. Results: Heterotrophic plate counts of DSW in the spring and summer season were investigated in the range of 0-550 and 0-800 CFU/mL, respectively. Total coliforms of these were 0-1,900 and 0-2,100 CFU/100mL, fecal coliforms were 0-600 and 0-550 CFU/100mL, and Escherichia coli were 0-7 and 0-326 MPN/100mL. The detection ratio of fecal pollution indicators and that of fecal coliforms increased to 58.6% in the summer from 12.5% in the spring and Escherichia coli increased to 51.4% from 4.7%. As a result of genetic analysis on the isolated bacteria, the genus of total coliforms was classified in the order of Enterobacter spp. 12.7%, Serratia spp. 7.3%, E. hermanii 6.4%, Rahnella spp. 5.5%, Hafnia spp. 4.5%, Escherichia coli 3.6%, Klebsiella spp. 3.6% in the spring season. In the summer season, it was classified in order of Klebsiella spp. 16.6%, Enterobacter spp. 13.0%, Escherichia coli 11.0%, Serratia spp. 8.6%, Raoultella spp. 7.0%, Kluyvera spp. 5.6% and Citrobacter spp. 3.0%. Conclusions: The increase of fecal pollution in summer indicates that special attention to drinking DSW is required.

• Korean Journal of Ecology and Environment
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• v.37 no.1 s.106
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• pp.26-35
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• 2004

We investigated the effect of physicochemical environmental factors on the community dynamics of phytoplanktons and bacteria at the Hoeya Dam Reservoir, a drinking water reservoir for Ulsan city. Water samples were collected and analyzed every two to four weeks at three sites along the reservoir from April to October, 2001. During the study period, the Secchi depths were between 0.4 and 3.5 m. At the surface layer of water column, temperature ranged 10.2 ~ $32.0^{\circ}C$, pH 7.3${\sim}$9.6, dissolved oxygen 5.5 ${\sim}$ 12.4 mg $L^{-1}$, $BOD_5$ 0.8 ${\sim}$ 5.0 mg $L^{-1}$, $COD_{Mn}$ 3.7 ${\sim}$ 10.0 mg $L^{-1}$, and Chl-a 8.9 ${\sim}$ 60.9 mg $m^{-3}$. At the bottom layer, temperature varied 7.2 ${\sim}$ $28.9^{\circ}C$, pH 7.1 ${\sim}$ 9.3, dissolved oxygen 0.6 ${\sim}$ 9.7 mg $L^{-1}$, $BOD_5$ 0.8 ${\sim}$ 4.5 mg $L^{-1}$, $COD_{Mn}$ 3.9 ${\sim}$ 10.0 mg $L^{-1}$, and Chl-a 4.3 ${\sim}$ 81.9 mg $m^{-3}$. The numbers of phytoplanktons were 7.4${\pm}10^2{\sim}2.6{\pm}10^5$ cells $mL^{-1}$ at surface and 2.5${\pm}10^2{\sim}2.4{\pm}10^4$ cells $mL^{-1}$ at bottom, and were positively correlated with water temperature and Chl- a concentration. Genus Stephanodiscus and genus Oscillatoria dominated on April and on May, respectively. Cyanobacterial blooms of Aphanizomenon, Microcystis, Anabaena were observed from June to early September, and thereafter Stephanodiscus and Aulacoseiral dominated again. Total microbial counts ranged 1.73${\pm}10^4{\sim}1.68{\pm}10^5$ cells $mL^{-1}$, and were positively correlated with water temperature and phytoplankton counts at surface water. Heterotrophic plate counts (HPCs) ranged 30${\sim}4.1{\pm}10^3$ CFU $mL^{-1}$, and were positively correlated with $BOD_5$ and $NO^3\;^-$-N concentration at bottom water. Unlike the total microbial counts, the numbers of fecal coliforms and fecal streptococci as well as HPCs were higher at the bottom than the surface layer and were highest at the upper a site among the three sampling sites. Since the concentrations of fecal coliforms and streptococci were still high at the bottom of site c, where intake for water treatment plant is located, it appeared that special management of water treatment processes may be needed especially after strong rainfall.