• Title/Summary/Keyword: MPN method

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Survival of Sanitary Indicative Bacteria Inoculated in Fish Muscle Homogenates during Freezing and Frozen Storage (어육에 접종한 위생지표세균의 동결저장중 변화)

  • CHOI Jong Duck;CHANG Dong Suck;KIM Young Man
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.19 no.4
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    • pp.356-362
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    • 1986
  • This experiment was designed to evaluate effects of freezing and frozen storage on survival of sanitary indicative bacteria in seafoods. Culture of bacteria such as Escherichia coli type I, Citrobacter freundii type I, Klebsiella aerogenes type I and Streptococcus faecalis was inoculated into homogenates of pollack, shrimp, and sardine frozen in a contact plate freezer at $-40^{\circ}C$ and chest freezer at $-20^{\circ}C$, stored at $-20^{\circ}C$, and then survival of the inoculated bacteria was determined over a period of 95 days. Coliform group was highly sensitive to freezing and frozen storage showing survival of about $2\%$ after 95 days of frozen storage at $-20^{\circ}C$, whereas Streptococcus faecalis was relatively resistant with $20\%$ survival rate. The sanitary indicative bacteria count was rapidly decreased in the early stage of frozen storage revealing 90 to $95\%$ loss of coliform group and 40 to $70\%$ loss in case of Streptococcus faecalis after 10 days storage. In determining recovery rate, most probable number (MPN) method gave more reproducible recovery of the tested strain than did the selected agar plate method.

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Rapid Enumeration of Listeria monocytogenes in Pork Meat Using Competitive PCR

  • Lim, Hyung-Kun;Hong, Chong-Hae;Choi, Weon-Sang
    • Food Science and Biotechnology
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    • v.14 no.3
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    • pp.387-391
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    • 2005
  • Competitive polymerase chain reaction (cPCR) was used to develop a direct enumeration method of Listeria monocytogenes in pork meat. Pork meat was artificially inoculated with L. monocytogenes and DNA was extracted using guanidine thiocyanate-phenol-chloroform and subjected to PCR amplification. Sixteen primer sets for L. monocytogenes hlyA gene were tested for sensitive detection and the DG69/DG74 primer set was selected. The detection limit achieved with this primer set was as low as 860 colony-forming units (cfu) per 0.1 g of pork meat. When the samples were cultured at $30^{\circ}C$ for 16 hr in Brain Heart Infusion (BHI) medium, even a single bacterium could be detected with this primer set by PCR. For cPCR, the hlyA gene, which features a 148 bp-deletion, was cloned in the pGEM-4Z vector. A known amount of competitor DNA which has the same primer binding sites was co-amplified with L. monocytogenes total DNA from the artificially inoculated pork meat. The cell-number determined by cPCR was approximately equal to cfu from the Most Probable Number (MPN) method. The whole procedure took only 5 hr.

Comparison of an Automated Technique TEMPO with Direct Plate Count Method for the Enumeration of Staphylococcus aureus (황색포도상구균의 TEMPO STA와 표준 평판 배지를 이용한 정량분석법 비교)

  • Cho, Yong Sun;Lee, Da-Yeon;Lee, Joo-Young;Wang, Hae-Jin;Shin, Dong-Bin
    • Journal of Food Hygiene and Safety
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    • v.28 no.3
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    • pp.252-257
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    • 2013
  • In this study the automated technique TEMPO STA method, a new automated enumeration based on MPN method, was compared to the direct plate count method by testing various ready-to-eat compound food products. Artificially inoculated food samples with Staphylococcus aureus - Bibimbap, Bibimnaengmyeon, Mulnaengmyeon, Kimmaki, Salad, Japchae, and Sushi- were tested. Statistical analysis of the results showed above 99% in agreement between the two enumeration methods. Also, we monitored 466 various ready-to-eat compound food products samples. The measure uncertainties of the two methods is also familiar, while TEMPO STA automated method is simpler, less time-consuming, more effective and more stable.

Detection of Enterovituses from Surface Water by Combined Cell Culture-PCR (지표수로부터 세포배양-연계 PCR법에 의한 장바이러스의 검출)

  • 정은영;정종문;류재익;신판세;전홍기;장경립
    • Journal of Life Science
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    • v.10 no.5
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    • pp.484-489
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    • 2000
  • Enterovirues may cause gastrointestinal symptoms, cold, and fever, mainly in young children. They are also recognized as important agents in acute infections of the central nervous system such as meningitis and encephalitis, and in subacute and chronic infections of the cardiovascular system such as pericarditis, myocarditis and cardiomyopathy. They also can lead to postviral fatigue syndrome. For the detection of enteroviruses from the environmental samples, the combined cell culture-polymerase chain reaction (CC-PCR) technique was employed. In contrast to EPA standard method which mainly depends on the cell culture, it involved the use of cell culture, followed by PCR to improve the sensitivity and the accuracy of the test. According to the results of survey, from 1999 to 2000, for the presence of enteroviruses in the surface water samples from Nak-dong river, four out of twelve samples were positive for viruses. The titer of viruses in the surface water was ranged from 25 to 250 MPN. All of the viruses isolated were poliovirus type I with 98% nucleotide sequence homology. The result also clearly suggests the seasonal difference in the distribution of the waterborne enteroviruses in surface water because most of the viruses were mainly detected from the summer through the early autumn.

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A Study on Bacterial Contamination of Ice Tea Sold on the Street in Seoul Area (路上販賣冷茶의 세균오염에 관한 조사연구)

  • Jang, Jae Seon
    • Journal of Environmental Health Sciences
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    • v.12 no.2
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    • pp.39-48
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    • 1986
  • This study was carried out to investigate the bacterial contamination of Ice Tea sold on the l street in Seoul area. For this, study 81 samples were collected on the street from July to September, 1985 and were examined on the following items. 1. Degree of bacterial contamination. 2. The relation of the occurrence of fecal coliform and salmonella. 3. The change of bacterial contamination in Ice Tea against temperature. As the results of this study, the following conclusion were obtained. 1. The mean count of total viable bacteria by standard plate count was $6.5{\times}10^3$/ml, the mean count of total coliform and fecal coliform by MPN method were $3.4{\times}10^2$/100ml, 5.5/100ml and those of fecal streptococci was $3.2{\times}10^2$/100ml. 2. The mean count of Staphylococcus aureus was 10.5/ml, the isolated rate of salmonella was 7.41%. 3. In relation to the occurrence of fecal colfform and salmonella, salmonella isolated that for values above $10^2$ fecal coliform 100ml. 4. In the change of bacterial contamination in Ice Tea against temperature, the number of total coliform and fecal coliform increased at $25{\circ}$C, decreased at $4{\circ}$C, but fecal streptococci increased at $25{\circ}$C and $4{\circ}$C.

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Effect of $Cd^{2+}$ and $Cu^{2+}$ on the Growth of a Methanogen and a Sulfate-Reducing Bacterium isolated from sea-based landfill (해안폐기물매립지로부터 분리한 메탄생성균과 환산염 환원균의 $Cd^{2+}$$Cu^{2+}$에 대한 감수성 검토)

  • Chang, Young-Cheol;Jeong, Kweon;Jeon, Eun-Mi;Bae, Il-Sang;Kim, Kwang-Jin
    • Journal of Environmental Health Sciences
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    • v.26 no.4
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    • pp.65-74
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    • 2000
  • The sensitivity of a mehtanogen and sulfate-reducing bacterium isolated from a sea-based landfill site Cd$^{2+}$ and CU$^{2+}$ was studied. Methanogens and sulfate-reducing bacteria in leachates of the waste disposal site were enumerated using the MPN method. Methanobacterium thermoautotrophicum KHT, isolated from the leachate, could not grow at 0.5 mM Cd$^{2+}$ or 1.0 mM CU$^{2+}$. Desulfotomaculum sp. RHT, isolated from the same leachate, was able to insolubilization 3.0 mM Cd$^{2+}$ or 2.0 mM CU$^{2+}$ by production of hydrogen sulfide. When strains KHT and RHT were cultured together in the presence of the heavy metals, strain KHT could grow at high heavy metal concentrations after insolubilization of the metals by strain RHT. strain RHT.

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Effects of $PCO_2$ on Methane Production Rate and Matter degradation in Anaerobic Digestion (혐기성소화의 물질분해 및 메탄생성에 대한 $CO_2$ 분압의 영향)

  • 이국의;김영철;서명교
    • Journal of Environmental Health Sciences
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    • v.26 no.2
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    • pp.59-66
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    • 2000
  • Effects of carbon dioxide partial pressure(PCO2) on bacterial population, methane production rate and matter degradation in anaerobic digestion were investigated by using anaerobic chemostat type reactors at 35$\pm$1$^{\circ}C$, at the HRT of 7 days. At PCO2 of 0.5 atm, the specific methane production rate and specific substrate removal rate reached the maximum rates. The methane production rates in the reactors fed by mixed substrate were 26% higher than those obtained under the controlled condition. The number of acetate consuming methanogenic bacteria enumerated by the MPN(most probable number) method, decreased when PCO2 exceeded 0.7 atm. Hydrogen consuming methanogenic bacteria and homoacetogenic bacteria increased as PCO2 increased from 0.1 to 0.6 atm, however, decreased slightly at PCO2 above 0.7 atm. The number of hydrolytic bacteria, sulfate-reducing bacteria and H2-producing acetogenic bacterial were not much influenced by the change of PCO2. The potential methanogenic activity reached the maximum at PCO2 0.5 atm, however, decreased significantly when PCO2 exceeded 0.7 atm, would depend on free PCO2 concentration in solution.

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Evaluation of Dry Rehydratable Film Method for Enumeration of Microorganisms in Meat, Dairy and Fishery Products (축.수산식품 중 미생물 분석을 위한 건조필름법 평가)

  • Cho, Mi-Hee;Bae, Eun-Kyung;Ha, Sang-Do;Park, Young-Seo;Mok, Chul-Kyoon;Hong, Kwan-Pyo;Kim, Sang-Phil;Park, Ji-Yong
    • Korean Journal of Food Science and Technology
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    • v.37 no.2
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    • pp.294-300
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    • 2005
  • Contents of total aerobic bacteria, coliform, Escherichia coli, yeast, mold, and Staphylococcus aureus in meat, dairy, and fishery products were analyzed by dry rehydratable film method using 3M $Petrifilm^{TM}$ and compared against those obtained through conventional method. Two methods showed high correlations of 0.990-0.999, 0.975-0.999, 0.979-0.987, 0.978-0.984, and 0.999 for total aerobic bacteria, yeast and mold, coliform, E. coli, and S. aureus, respectively; therefore, dry rehydratable film method using 3M $Petrifilm^{TM}$ offers acceptable alternative to conventional method for enumeration of microorganisms in meat, dairy, and fishery products.

Effects of Soil Environmental Conditions on the Decomposition Rate of Insecticide Fenitrothion in Flooded Soils (담수토양중(湛水土壤中)에 있어서 살충제(殺蟲劑) fenitrothion의 분해속도(分解速度)에 미치는 각종(各種) 토양환경조건(土壤環境條件)의 영향)

  • Moon, Young-Hee
    • Korean Journal of Environmental Agriculture
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    • v.9 no.1
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    • pp.1-8
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    • 1990
  • The effects of soil environmental conditions on the degradation rates of fenitrothion(O-O-dimethyl O-4-nitro-m-tolyl phosphorothioate) in soils under flooded conditions were examined in the laboratory. Fenitrothion was degraded rapidly and the half life period was within 4 days. Furthermore the degradation was mere rapid under flooded conditions than under upland conditions. The decomposition rate was varied with soils and soil temperatures. Fenitrothion degraded more slowly at 30ppm than at l0ppm. Repeated applications of fenitrothion in soils accelerated the degradation rates. The degradation remarkably increased with amendment of rice straw. However, degradation rates ,were virtually unaffected by the addition of the mixed-fertilizer, the fungicide IBP and the herbicide butachlor. The population of fenitrothion-degrading microbes, which were counted by MPN method, always corresponded with the degradation rates in the soils.

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Relationship between the Organic Content, Heavy Metal Concentration and Anaerobic Respiration Bacteria in the Sediments of Shiwha-ho (시화호 저니(Sediment)에서의 유기물 및 중금속 농도와 혐기성호흡세균과의 상관관계)

  • 현문식;장인섭;박형수;김병홍;김형주;이홍금;권개경
    • Microbiology and Biotechnology Letters
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    • v.27 no.3
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    • pp.252-259
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    • 1999
  • Anoxic sediments collected from Shiwha-ho area were used to find the relationship between the heavy-metal, organic content and anaerobic respiration bacteria by most probable number (MPN) method. Analysis of the sediments showed that COD content was higher in the sediments collected from Ansan-cheon and Shiwha-ho than those collected from sea area nearby. Particularly noticeable was the fact that heavy metal concentration was much higher in the sediments of Shiwha-ho area contaminated by heavy-metal, although they were rich in electron donor and electron acceptor for Fe(III)-reducing bacteria using lactate as an electron donor was in the range of 1.1$\times$106-4.6$\times$107MPNs/ml in the sediments collected from the sea-side of the lake, which were lower in heavy-methal concentration and higher in Fe-Mn content than those from other region. The number of Fe(III)-reducing bacteria using acetate as an electron donor was in the rang eof 4.3$\times$102-8.1$\times$105MPNs/ml in the same sediments. Chromate-reducing bacteria were more populated(4.6$\times$104-8.1$\times$105MPNs/ml) in the sediments contaminated by heavy metals. The number of sulfate-reducing bacteria wee counted in the sediments collected from the more contaminate inner-side than those from the sea-side of the lake.

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