• Title/Summary/Keyword: direct plate count method

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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.

Application of qDVC Method for Measuring Viable Cells in Lakes (호수 생태계에서 살아있는 세균을 측정하기 위한 qDVC 방법의 적용)

  • Kim, Mi-Ree;Seo, Eun-Young;Choi, Seung-Ik;Ahn, Tae-Seok
    • Korean Journal of Microbiology
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    • v.42 no.3
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    • pp.205-209
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    • 2006
  • For measuring the viable cells in lakes, quantitative direct viable count (qDVC) method is applied. In the qDVC process, the final concentration of glycine is fixed as 2%. For confirming the effectiveness of qDVC for enumerating the viable cells, the viable bacterial numbers were measured by plate count, CTC reduction method and qDVC method at 5 different lakes. Among these 3 methods, the bacterial numbers by qDVC is $2.4{\sim}6.0$ times higher than those by the other 2 methods. And by the qDVC method, the viable cells were easily discriminated from dead or dormant cells.

Direct and Quantitative Analysis of Salmonella enterica Serovar Typhimurium Using Real-Time PCR from Artificially Contaminated Chicken Meat

  • Park, Hee-Jin;Kim, Hyun-Joong;Park, Si-Hong;Shin, Eun-Gyeong;Kim, Jae-Hwan;Kim, Hae-Yeong
    • Journal of Microbiology and Biotechnology
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    • v.18 no.8
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    • pp.1453-1458
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    • 2008
  • For quantitative PCR assay of Salmonella enterica serovar Typhimurium in food samples, a real-time PCR method was developed, based on DNA genome equivalent. Specific primers and probe designed based on the STM4497 gene of S. Typhimurium LT2 showed the specificity to S. Typhimurium. Threshold cycle (Ct) values of real-time PCR were obtained from a quantitative standard curve with genomic DNA of Salmonella Typhimurium. In addition, the recovery of S. Typhimurium inoculated artificially to chicken samples with $4.5{\times}10^5$ to 4.5 CFU/ml was evaluated by using real-time PCR and plate-count methods. Result showed that the number of cells calculated from the real-time PCR method had good correlation with that of the plate-count method. This real-time PCR method could be applicable to the detection and quantification of S. Typhimurium in food samples.

Phylogenetic Characteristics of viable but Nonculturable Bacterial Populations in a Pine Mushroom (Tricholoma matsutake) Forest Soil (송이 자생군락 토양 내 난배양성 세균군집의 계통학적 특성)

  • Kim, Yun-Ji;Whang, Kyung-Sook
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.201-209
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    • 2007
  • The CFDA (6-carboxyfluorescein diacetate) direct viable count method and plate count (PC) method using conventional nutrient broth (NB) medium and $10^{-2}$ diluted NB (DNB) medium were applied to samples collected from Mt. Yongdoo In Andong, in an effect to determine the number of living bacteria pine mushroom forest soil. The number of living bacteria determined via plate count in NB medium comprised $5{\sim}8%$ of the CFDA direct viable count, and the bacteria in the DNB medium comprised $40{\sim}47%$. This result indicated that viable but nonculturable (VBNC) bacteria existed in the pine mushroom forest soil at a high percentage. The phylogenetic characteristics of the VBNC bacterial populations in the samples of pine mushroom (Tricholoma matsutake) forest soil were analyzed via the direct extraction of DNA and 16S rDNA-ARDRA. The 115 clones from pine mushroom forest soil were clustered into 31 different RFLP phylotypes by ARDRA. Based on the 16S rDNA sequences, the 31 ARDRA clusters were classified into 6 phylogenetic groups: ${\alpha}-,\;{\beta}-,\;{\gamma}-Proteobacteria$, Acidobacteria, Actinobacteria and Firmicutes. Among these bacterial populations, approximately 85% were classified as members of phylum Acidobacteria. The Acidobacteria phylum was shown to exist abundantly in the pine mushroom forest soil.

The Detection and a Quantitative Evaluation of Viable but Non-Culturable Soil Bacteria Using a Modified Direct Viable Count Method (변형된 DVC법을 이용한 난배양성 토양세균의 검출 및 정량적 평가)

  • 황경숙;양희찬;염곡효
    • Korean Journal of Microbiology
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    • v.39 no.3
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    • pp.181-186
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    • 2003
  • This study was performed to analyze quantitatively the number of living bacteria in forest soil samples collected from Mt. Keryong using improved direct viable count (DVC) and plate count (PC) methods. The number of living bacteria by DVC comprised 18~44% of the total direct count (TDC), whereas the number of living bacteria by PC was less than 1% of TDC. These results showed that viable but non-culturable (VBNC) bacteria existed in the soil with high percentages. Besides, DVC was proved to make it possible to make a quantitative detection of the VBNC bacteria. On the other hand, upon measuring the value from the conventional nutrient broth (NB) and $10^{-2}$ folded diluted nutrient broth (DNB), the values from the DNB showed 5 to 10 times higher than those from the conventional NB medium. These results indicate that oligotrophic bacterial groups, which could multiply in the low nutrient broth, abundantly exist in the soil ecosystem. It would also be possible to apply this kind of method to other substrate to make a quantitative detection of soil bacterial groups.

Screening of Sterilized Ramen Soup by DEFT/APC Method and Its Quality Properties as Affected by Irradiation (DEFT/APC 측정에 의한 시판 분말수프의 살균처리여부 확인 및 감마선 처리에 따른 품질특성 평가)

  • Ahn, Jae-Jun;Kim, Kwang-Hoon;Park, Sung-Hyun;Kwon, Joong-Ho
    • Korean Journal of Food Science and Technology
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    • v.41 no.5
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    • pp.515-521
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    • 2009
  • The DEFT (direct epifluorescent filter technique)/APC (aerobic plate count) test was utilized to screen powdered Ramen soup samples (RS-1, RS-2) whether or not they have been microbial-decontaminated. The initial microbial loads of commercially-packaged samples were log DEFT 6.46 (RS-1) and 7.05 (RS-2), but the viable bacterial counts were log APC 2.74 (RS-1) and 1.95 (RS-2), respectively; this finding showed that they have been already decontaminated by methods other than irradiation. The same samples were then subjected to gamma irradiation at 0, 5 and 10 kGy in order to evaluate the microbial and physicochemical changes during post-irradiation storage for 6 months under room conditions ($10{\pm}3^{\circ}C$). The DEFT count was constant in irradiated samples even at different doses, but APC decreased with dose increases; this implies that the log DEFT/APC increased in a linear fashion with dose. No coliforms, yeasts and molds were detected in any of the samples, whereas the initially detected aerobic bacteria ($5.49{\times}10^2CFU/g$) were inactivated by 5 kGy or more and no growth was observed during the subsequent storage period. The pH of RS-1 was reduced by irradiation (p<0.01), but increased (p<0.01) with increasing storage time. Irradiation induced a reduction in volatile basic nitrogen (VBN), whereas an increase in thiobarbituric acid (TBA) values was observed. The storage time proved more influential than irradiation up to 10 kGy in terms of changes in the VBN and Hunter’s color values of powdered Ramen soups.

Production of Standard Sample for Quality Control of Total Coliform (총대장균군 측정의 정도관리에 적합한 균주의 조제)

  • Kim, Ju-Young;Seo, Eun-Young;Kim, Mi-Ree;Jeon, Nam-Hui;Chung, Hyen-Mi;Kim, Myeong-Woon;Ahn, Tea-Seok
    • Korean Journal of Microbiology
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    • v.44 no.1
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    • pp.43-48
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    • 2008
  • Standard sample for quality control of total coliform measurement was procured by addition of nalidixic acid and cephalexin as bacteriostatic agents to Escherichia coli cultured broth. After making the standard sample, the number of E. coli was measured by fluorescence microscopic count method and plate count method by 12 hr interval. The numbers of E. coli remained unchanged for at least for 48 hr at room temperature which ranged from 3.5 to $4.2{\times}10^5\;cells/ml$ and from 1.0 to $1.2{\times}10^4\;CFU/ml$ by direct fluorescence microscopic count method and plate count method, respectively. This result suggests that microbial standard sample with bacteriostatic agents of nalidixic acid and cephalexin is usable for quantitative quality control.

Studies on the Hand Hygiene Practices of Food-Service Businesses Workers: A Comparison of Full-time and Part-time Workers (외식업소 종사자의 손 위생관리에 관한 연구)

  • Kim, Jong-Gyu;Park, Jeong-Yeong;Kim, Joong-Soon
    • Journal of Environmental Health Sciences
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    • v.39 no.1
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    • pp.71-82
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    • 2013
  • Objectives: This study was performed in order to investigate hand hygiene practices among food-service businesses employees based on the awareness of hand-washing and load of indicator bacteria on their hands. It focused on the comparison of full-time and part-time workers in food-service workplaces. Methods: A direct-interview questionnaire survey and microbiological analysis were carried out with sixty workers each. Samples for microbiological analysis were collected through a modified glove-juice method from the hands of the food-service workers and were analyzed for aerobic plate count, total coliform, fecal coliform, Escherichia coli, Staphylococcus aureus, and Salmonella spp. Microbiological analysis was done according to the Food Code of Korea. Results: Significant differences (p<0.01) were found in the survey between the full-time and part-time workers in hand-washing frequency, use of hand-washing agents, and hand-drying methods. More full-time workers responded to washing their hands after preparing food, after visiting outside, after handling raw materials, and before putting on gloves/when changing gloves than did part-time workers (p<0.05). No remarkable difference was found in bacterial load on the hands except in the aerobic plate count between the two groups. The detection of E. coli, S. aureus, and Salmonella spp. on the hands of some food-service workers in both groups revealed poor hand hygiene practices. Conclusions: The results of this study indicate that there is a need for training programs in order to improve hand hygiene practices and strict hand hygiene compliance by food-service workers.

A Study on the Hand Washing Awareness and Practices of Food-service Employees and the Load of Index Microorganisms on the Hands (조리종사자의 손 씻기 의식과 실천 및 손의 지표미생물 오염도에 관한 연구)

  • Park, Jeong-Yeong;Kim, Joong-Soon;Kim, Jong-Gyu
    • Journal of Environmental Health Sciences
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    • v.36 no.2
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    • pp.95-107
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    • 2010
  • Hand-washing is one of the major factors in personal hygiene and public health. This study was undertaken to investigate the hygienic behavior of food-service employees, focusing on awareness of hand washing, hand washing practices, and the load of index microorganisms (aerobic plate count, total and fecal coliforms, Escherichia coli, and Staphylococcus aureus) on the hands of food-service employees. A questionnaire survey completed by direct interview, direct observation of restrooms by the researcher and trained observers, and microbiological examination according to the Food Code of Korea were carried out. In the survey, a positive attitude toward hand washing compliance was reported; however, improper hand washing and poor hand hygiene of the food-service employees were seen under direct observation. Significant differences (p<0.05) were found between the questionnaire survey and the direct observations in hand washing compliance after using the toilet, duration of hand washing, use of hand washing agents, use of hand washing tools, washing of different parts of the hands, hand-drying method, temperature of water, and method of turning off the water. Samples taken from employees' hands before washing showed higher levels of bacteria than those taken during work and/or after washing (p<0.05). Poor hand washing practices were indicated by the positive results for total and fecal coliforms, E. coli, and S. aureus on the hands of some food-service employees. This study showed that there is a marked difference between the food-service employees' awareness of hand-washing and their actual hand-washing practices. The poor hand hygiene of and improper hand washing by the food-service employees should be addressed for improved food safety.

Recent Development of Rapid and Automation Technology for Food Microbiological Examination

  • Hiroshi Kurata
    • Proceedings of the Korean Society of Food Hygiene and Safety Conference
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    • 1996.06a
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    • pp.33-33
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    • 1996
  • Interests in the field of rapid methods and automation in microbiology have been growing steadily on an international scale in recent years. International meetings concerned this problem have been held in elsewhere in the world countries since the past twenty years. But, unfortunately in the field of microbial examination in food hygiene, this problem have not yet been developed so much as in the field of clinical microbiology. Today, I would like to introduce you here present aspects of rapid and automation technologies, those which are manly carrying in milk and meats industries. My illustration will be given recent improved technologies using automatic apparatus and instruments along with process of microbial count procedure. Recent direct microbiological counting system (ChemeScan \ulcorner) as real time ultrasensitive analysis created by Cheminex Ltd., France is now most evolutional instrument to provide direct microbial counts, down to one cell, within 30 minutes. The results from these evaluations how a good correlation between the ChemScan system and the standard plate count method. This system will be successful application for not only in the field of pharmacology but also food microbiology. In addition, current identification of microbes by sophisticated instruments suitable for food microbiology, one of which Biology is manual system (BIOLOG\ulcorner), provides reference-level capability at a modes price. For the manual system, the color reactions in the microplate are read by eye and manually keyed into personal computer. Species identification appears on the computer screen within seconds, along with biotype patterns, a list of closely related species, and other useful statistics. In present this is useful application for microbial ecology and epidemiological survey. RiboPrinter system newly produced by DuPont is now focusing among microbiologists in the world, and is one of the biggest microbial characterization system using a DNA-based approach. The technology analyzer is bacterial culture for its genetic fingerprint or riboprint pattern. Finally Bio-cellTracer system for automatic measurement of fungal growth and Fukitori-Maseter, a Surface Hygiene Monitoring Kit by using swabe procedure in food processing environment are briefly illustrated in this presentation.