• Title/Summary/Keyword: direct plate counting

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Effect of Diluent Salt Concentration and pH on the Enumeration of Vibrio parahaemolyticus by Direct Plating on Selective Agar

  • Lee, Jong-Kyung;Jung, Da-Wa;Yoon, Ki-Sun;Yoon, Sun-Kyung;Kwak, No-Seong
    • 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.

Growth inhibition of Saccharomyces cerevisiae by alternation current pulse (교류 펄스에 의한 Saccharomyces cerevisiae의 생장억제 효과)

  • 정지환;박현근;한홍의
    • Korean Journal of Microbiology
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    • v.25 no.3
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    • pp.249-253
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    • 1987
  • Effects of Ac pulse at low voltage on Saccharomyces cerevisiae were studied. The treatment of yeast suspensions contained 0.2m NaCl with 500mA for 350 sec at $40^{\circ}C$ was shown about 50% of lethality, whereas in the treatment of the same suspensions with 250mA for 250 sec at the temperature ($10^{\circ}C$) corresponding to about 10% of lethality, growth was completely inhibited instead of lethality. The effect of growth inhibition was due to occurrence of auxotrophic strains under experimental conditions. Detection of auxotrophic yeasts was done tentatively with the difference of the number of viable yeast cells between direct counting by methylene blue staining and plate-count on yeast morphology agar.

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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.

Studies on the Maintenance of Bioluminescence from Stored Photobacterium phosphoreum (Photobacterium phosphoreum의 생체발광 유지도에 관한 연구)

  • 김현숙;정성제;전억한
    • Microbiology and Biotechnology Letters
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    • v.28 no.2
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    • pp.117-123
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    • 2000
  • The object of this work is to improve the maintenance of bioluminescence from stored Photobacterium phosphoreum in a view of developing continuous monitoring system for pollutants. The long-term experiments were performed to determine the effect of storage temperature and immobilization on the maintenance of bioluminescence and viability of P. phosphoreum. A naturally luminescent bacterium, P. phosphoreum was starved in 2.5% Nael solution at $20^{\circ}C$, $4^{\circ}C$, -$20^{\circ}C$ and -$70^{\circ}C$ for 30 days. In vivo luminescence was measured by luminometry, and total cell concentrations and concentrations of culturable and viable cells were determined by acridine orange staining, dilution plate counting, and direct viable counting, respectively. The bioluminescence emission from cells stored at 4De was maintained up to 10 days while those with starved cells at other temperature ranges decreased to background level within 3 days. In terms of viability of cells, concentrations of cells stored at $20^{\circ}C$ were rapidly decreased as a result of cell lysis, leading to a drop in culturable and viable counts while cells stored at $4^{\circ}C$ was shown viable but nonculturable state during starvation. With immobilized cells on strontium alginate, the bioluminescence showed higher maintenance than free cells and decreased with count number of nonculturable cells.

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The Effect of Hwangryunhaedoktang on Proliferations of Various Human Cancer Cells (황련해독탕이 수종의 인간 암세포 증식에 미치는 영향)

  • Sung, Hyun Kyung;Min, Sang Yeon;Kim, Jang Hyun
    • The Journal of Pediatrics of Korean Medicine
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    • v.27 no.1
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    • pp.59-68
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    • 2013
  • Objectives The aim of this study is to investigate whether hwang-ryun-haedok-tang (HDT) affect proliferations of androgen-dependent LNCaP prostate cancer cells, androgen-independent PC-3, DU-145 prostate cancer cells, MCF-7 human breast cancer cells, A549, NCI-H292 human pulmonary cancer cells and K-562 human chronic myelogenous leukemia cells. Materials and Methods Effects of HDT on proliferations of each cancer cell line were investigated. 20,000 cells/well were plated in each well of 96-well culture plate. After 24 hrs, 0.01-10% of HDT in culture medium was added to cancer cells. The number of cells was counted by using SRB assay or direct cell counting method after 72 hours from drug treatment. Effect of baicalein or berebrine on proliferation was assessed according to the same method. Results (1) HDT inhibited proliferations of LNCaP, PC-3 and DU-145 prostate cancer cells. (2) HDT inhibited proliferation of MCF-7 breast cancer cells. (3) HDT also inhibited proliferations of A549, NCI-H292 pulmonary cancer cells and K-562 chronic myelogenous leukemia cells. (4) Baicalein and berberine also showed inhibitory effects on proliferations of prostate and breast cancer cells. Conclusion : HDT inhibited proliferations of human prostate, breast, pulmonary and blood cancer cells. These results suggest us the potential use of HDT as a chemopreventive or chemotherapeutic agent. Effect of HDT on human cancer should be further investigated using in vivo experimental models that can reflect pathophysiology of human cancer through another studies.

Application of Epifluorescence, Microscopy for Measurement of Bacterial Population in Water Supplies (용수중(用水中) 세균계수(細菌計數)를 위한 형광검경법(螢光檢鏡法)의 응용(應用))

  • Rhee, Young-Hwan;Shin, Seung-Yee
    • Korean Journal of Environmental Agriculture
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    • v.1 no.1
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    • pp.48-52
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    • 1982
  • Methods for the measurement of aquatic bacteria can be divided into two groups. The first group of these methods is based on the 'replicon' concept that live bacterial cells, when diluted and transferred to a suitable medium, produce colonies. These methods distinguish living from dead bacteria, but they massively underestimate bacterial numbers. The second group of enumeration methods uses visual counting technique using specific apparatus such as a microscope. These methods are generally direct and simple, but it is very hard to distinguish between live and dead bacteria and between small particle and bacteria. Recently developed technique in staining methods has provided a reliable method of visual determination of aquatic bacteria. This uses epifluorescence microscopy to measure the total bacterial population. In order to present the fluorescence microscopy as a new methodology for the determination of bacterial numbers in water supplies, data were obtained from chlorine and monochloramine doses added to samples. Total counts by fluorescence microscopy were compared with standard plate count method. The total number of bacteria in water supplies can be determined with fluorescence microscopy. This technique allows better resolution of small bacteria and differentiation of particle from bacteria. Chloramine was found to persist longer in natural waters and prevent bacterial regrowth.

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Interspecific Variation in the Protoplast Formation of the Genus Cellulomonas (Cellulomonas속 종간의 원형질체 형성조건의 차이에 대하여)

  • Lee, Eun-Ju;Bae, Moo
    • Korean Journal of Microbiology
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    • v.24 no.2
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    • pp.154-160
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    • 1986
  • In order to develope interspecific fusion of the genus Cellulomonas capable of assimilation cellulose, the optimun conditions for the protoplast formation was investigated to examine the susceptibility of cell wall, between different species of the same genus using scanning electron microscope. The variation in the susceptibilities of Cellulomonas sp. CS 1-1 and C. flavigena to lysozyme treatment were considerably remarkable, although they belong to the same genus. The rate of protoplast formation of CS1-1 was 99.9% being treated with lysozyme $(100{\mu}g/ml)$ for 30 minute and that of C. flavigena was about 80% being treated at the concentration of $600{\mu}g/ml$ of lysozyme for 6 hours. The susceptibility of cell wall to the lysozyme treatment on protoplast formation of the strain, CS1-1 seems not to be depend on the cultural periods of cells. On the contrary, that of C. flavigena was considerably depend on the periods. Cells of C. flavigena at mid exponential phase could be more efficiently converted to protoplast cells than those at late exponential phase be done. The rate of the protoplast formation was 95%, when cells of C. flavigena at mid exponential phase were treated with lysozyme $600{\mu}g/ml$ for 6 hours and observed by SEM. In the evalution of protoplast formation of the CS1-1 results of counting method in plate after osmotic shock treatment were similar to the results of the direct observation method by means of SEM. But in the case of C. flavigena the latter method was much more reliable than the former, because the differences between the number of spheroplasts and protoplasts were not able to figure out on conuting the number of protoplast after osmotic shock tretment.

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Adhesion Characteristics and the High Pressure Resistance of Biofilm Bacteria in Seawater Reverse Osmosis Desalination Process (역삼투 해수담수화 공정 내 바이오필름 형성 미생물의 부착 및 고압내성 특성)

  • Jung, Ji-Yeon;Lee, Jin-Wook;Kim, Sung-Youn;Kim, In-S.
    • Journal of Korean Society of Environmental Engineers
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    • v.31 no.1
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    • pp.51-57
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    • 2009
  • Biofouling in seawater reverse osmosis (SWRO) desalination process causes many problems such as flux decline, biodegradation of membrane, increased cleaning time, and increased energy consumption and operational cost. Therefore biofouling is considered as the most critical problem in system operation. To control biofouling in early stage, detection of the most problematic bacteria causing biofouling is required. In this study, six model bacteria were chosen; Bacillus sp., Flavobacterium sp., Mycobacterium sp., Pseudomonas aeruginosa, Pseudomonas fluorescens, and Rhodobacter sp. based on report in the literature and phylogenetic analysis of seawater intake and fouled RO membrane. The adhesion to RO membrane, the high pressure resistance, and the hydrophobicity of the six model bacteria were examined to find out their fouling potential. Rhodobacter sp. and Mycobacterium sp. were found to attach very well to RO membrane surface compared to others used in this study. The test of hydrophobicity revealed that the bacteria which have high hydrophobicity or similar contact angle with RO membrane ($63^{\circ}$ of contact angle) easily attached to RO membrane surface. P. aeruginosa which is highly hydrophilic ($23.07^{\circ}$ of contact angle) showed the least adhesion characteristic among six model bacteria. After applying a pressure of 800 psi to the sample, Rhodobacter sp. was found to show the highest reduction rate; with 59-73% of the cells removed from the membrane under pressure. P. fluorescens on the other hand analyzed as the most pressure resistant bacteria among six model bacteria. The difference between reduction rates using direct counting and plate counting indicates that the viability of each model bacteria was affected significantly from the high pressure. Most cells subjected to high pressure were unable to form colonies even thought they maintained their structural integrity.