• Microbiology and Biotechnology Letters
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• v.24 no.3
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• pp.364-370
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• 1996

Sediments collected from the Jungnang-cheon and its tributaries were used to enumerate anaerobic bacteria by most probable number (MPN) methods. A simple method was developed to detect ferrous ion in the culture fluid in order to count the number of iron ion reducers, and sulfate-reducing bacteria (SRB) and methanogens were detected by the presence of FeS precipitate in the culture or methane in the head space, respectively. The numbers of iron reducer was in the range of 10$^{7}$ - 10$^{8}$ /g in the sediment of the stream containing higher organic content than the tributaries. The sediments of tributaries were analyzed to contain iron reducers less than 10$^{7}$ cells/g. With one exception the numbers of SRB and methanogens were less than 10$^{3}$ cells/g in the sediment. From these results it is concluded that organics in the sediment support the growth of iron reducers, which out-compete SRB and methanogens.

• Journal of Korean Society on Water Environment
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• v.22 no.6
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• pp.1052-1059
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• 2006

For monitoring the fecal pollution at Nak-Dong River, one of the eutrophicated rivers, the differences between total coliforms (TC) and fecal coliforms (FC) using both of membrane filtration (MF)/MPN method, and also fecal streptococcus (FS) by MF-method was investigated. To evaluate the correlation between TC, FC, and FS statistical analyses were performed by using Minitab. And a part of the presumptive TC/FC and background colonies was purified and identified using API 20E kit (Biomeriux). As results, most (89%) of presumptive FC by MF was identified as Escherichia coli while only 14% (MPN) and 11% (MF) of TC were identified as E. coli. Furthermore, FC by MF was correlated significantly with other fecal indicators (TC/FS by MF and FC by MPN), while TC by MPN was not correlated with any other indicators. Thus, the detection of FC by MF-method may be the most reasonable for monitoring the fecal pollution.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.26 no.3
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• pp.205-213
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• 1993

The APHA-MPN procedure is the only officially accepted method for classifying shellfish growing areas in U. S. A. The method estimates the levels of fecal coliforms and E. coli, indicators of the sanitary quality of environmental waters. However, the MPN has several disadvantages requiring far more time, labor and expense for assay, as well as providing relatively poor precision. Several membrane filtration procedures have been developed to enumerate these indicators in waters. Of these, the mTEC technique has been shown to provide recoveries of fecal coliforms and E. coli comparable to those of the MPN method. In an abbreviated sanitary survey for Greenwich Bay in Rhode Island, U. S. A., classified as an approved shellfish growing area, the mTEC and conventional MPN methods were again compared for their recoveries of the indicator bacteria. It was found that the recoveries of fecal coliforms and E. coli provided by the mTEC technique are 1.08 and 1.27 times higher than those produced by MPN for water monitoring, respectively, and that the membrane filtration method appears to be a possible alternative to APHA-MPN.

• Journal of Korean Society on Water Environment
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• v.16 no.4
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• pp.541-554
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• 2000

Experimental study of anaerobic digestion process combined with and without the submerged separation membrane was investigated by using laboratory-scale reactor at the hydraulic retention time of 0.5 day and 1.0 day. The removal efficiencies of carbohydrate at the digester without and with membrane were 84.4 to 86.8 % and 99.6 to 99.7 %, respectively, and the methane gas recovery efficiencies were 0.4 to 1.2 % and 12.3 to 28.7 %. According to the measurement by the most probable numbers method. the numbers of various groups of bacteria in the digesters with membrane were enumerated in the following ranges ; acidogens : $1.7{\times}10^9$ to $5.0{\times}10^9MPN/m{\ell}$, homoacetogens : $5.0{\times}10^7$ to $2.4{\times}10^8MPN/m{\ell}$, $H_2$-utilizing methanogens : $1.3{\times}10^7$ to $9.2{\times}10^8MPN/m{\ell}$, and acetate-utilizing methanogens : $2.3{\times}10^6$ to $2.0{\times}10^8MPN/m{\ell}$. The number of methanogens at the digester with membrane increased by approximately $10^2$ to $10^4$ times in comparison with that of the digester without membrane.

• Journal of Microbiology and Biotechnology
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• v.20 no.2
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• pp.391-396
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• 2010

The purpose of this study was to investigate the biosafety of Vibrio parahaemolyticus in raw salad vegetables at wet markets and supermarkets in Malaysia. A combination of the most probable number-polymerase chain reaction (MPN-PCR) method was applied to detect the presence of V. parahaemolyticus and to enumerate their density in the food samples. The study analyzed 276 samples of common vegetables eaten raw in Malaysia (Wild cosmos=8; Japanese parsley=21; Cabbage=30; Lettuce=16; Indian pennywort=17; Carrot=31; Sweet potato=29; Tomato=38; Cucumber=28; Four-winged bean=26; Long bean=32). The samples were purchased from two supermarkets (A and B) and two wet markets (C and D). The occurrence of V. parahaemolyticus detected was 20.65%, with a higher frequency of V. parahaemolyticus in vegetables obtained from wet markets (Wet market C=27.27%; Wet Market D=32.05%) compared with supermarkets (Supermarket A=1.64%; Supermarket B=16.67%). V. parahaemolyticus was most prevalent in Indian pennywort (41.18%). The density of V. parahaemolyticus in all the samples ranged from <3 up to >2,400 MPN/g, mostly <3 MPN/g concentration. Raw vegetables from wet markets contained higher levels of V. parahaemolyticus compared with supermarkets. Although V. parahaemolyticus was present in raw vegetables, its numbers were low. The results suggest that raw vegetables act as a transmission route for V. parahaemolyticus. This study will be the first biosafety assessment of V. parahaemolyticus in raw vegetables in Malaysia.

• Food Science and Biotechnology
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• v.15 no.6
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• pp.866-870
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• 2006

The maintenance of physiological activity during dilution is very critical for the accurate enumeration of Vibrio spp. in marine samples. We investigated the effect of various diluents on the recovery of Vibrio parahaemolyticus using the direct plate counting and most probable number (MPN) methods. The effects of NaCl (0.85 and 3%) and pH (from 6.6 to 7.4) in diluents based on distilled water or phosphate buffered saline (PBS) were evaluated with three V. parahaemolyticus strains. PBS-3% NaCl (pH 6.6), as opposed to PBS, was the most effective diluent at maintaining viable cell numbers up to 2 log CFU/g during dilution for direct plate counting using on thiosulfate citrate bile salts sucrose (TCBS) selective agar, as well as minimizing the difference in cell numbers between TCBS and non-selective nutrient agar. It also increased counts of V parahaemolyticus inoculated into oysters relative to PBS (p<0.01), suggesting that PBS-3% NaCl (PH 6.6) can reduce the problem of underestimating V. parhaemolyticus counts using PBS alone.

• Journal of Korean Society of Environmental Engineers
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• v.22 no.3
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• pp.417-427
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• 2000

The substrate metabolism and bacterial population in an anaerobic digestion with the submerged separation membrane were investigated by using a laboratory-scale reactor at the hydraulic retention time(HRT) 1.0 and 0.5 day. The removal efficiencies of carbohydrate at the HRT 1.0 and 0.5 day were 99.8~99.9% and 98.0~99.6%, respectively. After the 58 days, the mixed liquor volatile suspended solids(MLVSS) concentration at the HRT 1.0 and 0.5 day were approximately 6,050 and 7,750 mg/L, respectively. According to the measurement by the most probable number(MPN) method, the numbers of acidogenic bacteria, $H_2$-utilizing and acetate-utilizing methc.nogenic bacteria were found to be $10^9$, $10^7{\sim}10^8$ and $10^6{\sim}10^8MPN/mL$, respectively. The composition of $CH_4$ in the produced gas was 46~50%. It is suggested that sulfate-reducing bacteria $10^7{\sim}10^8MPN/mL$ play an important role in producing $H_2$ and acetate in sulfate-depleted environment.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.41 no.3
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• pp.155-162
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• 2008

The seawater in the Taean area was surveyed to evaluate the conditions of the bay and compliance with the bacteriological criteria for a designated area for shellfish production for export. Samples of seawater were collected monthly at 34 sampling stations established in the survey area from January 2002 to December 2004. The bacterial density in the coastal area close to a pollution source located to the northeast of the survey area was higher than in the open sea to the west. The bacteriological counts in the water did not change with 16.5mm of rainfall, but increased abruptly after 65.4mm of rainfall. The total coliform and fecal coliform most probable numbers (MPNs) of 1,224 seawater samples in the survey area were <1.8-2,400 and <1.8-790 MPN/100 mL, respectively. The geometric mean and estimated 90th percentile of total coliforms were 1.9-3.4 and 2.7-26.3 MPN/100mL, respectively, and for fecal coliforms were <1.8-2.6 and 1.8-12.0 MPN/100mL, respectively. The bacteriological water quality in the Taean seawater area met the National Shellfish Sanitation Program criteria for an approved area and the Korea Shellfish Sanitation Program criteria for a designated area for shellfish production for export.

• Journal of the Korean Society of Food Science and Nutrition
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• v.12 no.1
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• pp.19-26
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• 1983

The purpose of the present study is to obtain some fundamental data for hygienic control of sanitary indicative bacteria of sea water in fish preservatory tanks at popular swimming beaches in Busan including Songdo, Kwanganri, Haeundae and Songjeong from July to August in 1982. The geometric mean values of coliform group and fecal coliform MPN (most probable number) of sea water of Kwanganri were 27,300/100ml and 8,700/100ml respectively, which showed the highest population of the study regions. MPN's of the sea water of beaches and fish preservatory tanks showed 21,000/100ml and 8,800/100ml respectively at Kwanganri, which were also the highest numbers. The degree of the hygienic bacteriological populations of both the seawater and that of the fish tanks at a same region showed similar trend. The pollution caused by coliform group generally increased as viable cell counts increased. The types of coliform organisms isolated from seawater in fish presorvatory tanks by IMViC reaction were 42% Escherichia coli group, 2% Citrobacter freundii group, 44% Klebsiella aerogenes group and 12% others.

• Journal of fish pathology
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• v.6 no.2
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• pp.205-218
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• 1993

On the development of hirame(Paratichtys olivaceus) culture, outbreak of scuticociliata infection was reported to cause severe damage in Japan. To establish effective measures for isolation and cultivation of this ciliate, we tried to culture this pathogenic ciliate using medium for bacteria and fish cell lines in vitro. Scuticociliata from the brain tissues of infected fish was aseptically inoculated to CHSE-214 cells cultured in MEM-10 without antibiotic. Scuticociliata grew well and the number of ciliate reached $10^6\;cells/ml$ at temperatures of $15^{\circ}C$ to $20^{\circ}C$ for 10d. The number of ciliate cultured in the cell lines is 10 times higher than the numbers cultured in the liquid medium alone. This ciliata could be cloned by dilution method. Scuticociliata isolated could grow well on 42 different cell lines that were established from marine fish, warm freshwater fish, and salmonids. This ciliate could be preserved in liquid nitrogen for more than 6 months. Subsequently, we observed the optimal temperature and salinity for growth, and tested the sensitivities of this organism to formaldehyde, flagyl(Metronidazole), Ekuteshin(Combination compound of sulfamonometoxin and ormethoprim), and ozonixation. Optimal temperature for growth was $25^{\circ}C$ and salinity was 1.0 to 1.5%. Washed scuticociliata was killed by formaldehyde at the concentration of 50ppm for 10min, but was not completely killed even at a high concentration of 400ppm for 20min in MEM-5. Flagyl and Ekuteshin can inhibit the growth of scuticociliata at the concentration of 1,000 and $100{\mu}g/ml$ in MEM-10, respectively. More than 99% of this scuticociliata could be killed by ozonization at a dose equivalent to $1.0mg/\ell$ oxidant for 30sec in sea water. Isolated scuticociliata showed the pathogenicity to the cultured hirame by artificial infection(I. P. injection, $10^5\;cells$/fish). The number of scuticociliata in the water could be counted by most probable number(MPN) method using tissue culture, and the minimum detectable number was $1.8\;cells/\ell$. The number in the reservoir tank for water supply to the culture tank was 110 cells/l. After cleaning by elimination of the sediments from of the reservoir tank and disinfected with formaldehyde, number of scuticociliata decreased and was counted less than $1.8\;cells/\ell$ and infection rate of cultured hirame was decreased.