• Journal of Dairy Science and Biotechnology
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• 제41권4호
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• pp.157-162
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• 2023

In the field of microbiology, numerous types of bacteria live dormant to survive stresses such as pasteurization and antibiotics. Some bacteria become 'persisters' by inactivating their ribosomes, allowing them to 'sleep' through stress and revive when the stress has been removed. Under stress, some cells morph into hollow, lifeless structures known as 'cell shells.' In microbiology, these cells have been confused with viable cells in the 'viable but non-culturable cells' phenomenon. Therefore, this review addressed the concept that when revival occurs, the always-viable persister cells revive, instead of the dead cell husks.

• Korean Journal of Microbiology
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• 제42권3호
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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.

• Journal of Microbiology and Biotechnology
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• 제23권12호
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• pp.1708-1716
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• 2013

Conventional molecular detection methods cannot distinguish between viable and dead Escherichia coli O157 cells. In this study, the loop-mediated isothermal amplification (LAMP) method combined with propidium monoazide (PMA) treatment was developed to selectively detect viable E. coli O157 cells. Four primers, including outer primers and inner primers, were specially designed for the recognition of six distinct sequences on the serogroups (O157) of the specific rfbE gene of the E. coli O157 genome. PMA selectively penetrated through the compromised cell membranes and intercalated into DNA. Amplification of DNA from dead cells was completely inhibited by $3.0{\mu}g/ml$ PMA, whereas the DNA derived from viable cells was amplified remarkably within 1 h by PMA-LAMP. Exhibiting high sensitivity and specificity, PMA-LAMP is a suitable method for evaluating the inactivation efficacy of slightly acidic electrolyzed water in broth. PMA-LAMP can selectively detect viable E. coli O157 cells. This study offers a novel molecular detection method to distinguish between viable and dead E. coli O157 cells.

• Korean Journal of Fisheries and Aquatic Sciences
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• 제29권6호
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• pp.882-887
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• 1996

Production of yeast diet has been carried out in batch fermenters under optimum culture conditions. The fermentation of Candida utilis on a $2\%$ complex medium resulted in 1.22 g/L/h productivity and $65\times10^8$ viable cells/ml, and the addition of $15{\mu}M$ zinc to the medium increased both the productivity and the number of viable cells just a little more. In the case of the fermentation of Kluyveromyces fragilis, the highest value of the biomass productivity, 1.94 g/L/h, was obtained on a $2.5\%$ fructose medium with $70\times10^8$ viable cells/ml, and $1\%$ peptone was found to be a growth factor in this fermentation. When $3.5\%$ NaCl was added to the given medium, both the biomass productivity and the number of viable cells decreased significantly in each fermentation, but this may be considered to preserve yeast diet long without osmotic lysis.

• Journal of Microbiology and Biotechnology
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• 제17권9호
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• pp.1558-1562
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• 2007

To elucidate the influence of pipe materials on the VBNC (viable but nonculturable) state and bacterial numbers in drinking water, biofilm and effluent from stainless steel, galvanized iron, and polyvinyl chloride pipe wafers were analyzed. Although no HPC (heterotrophic plate count) was detected in the chlorinated influent of the model system, a DVC (direct viable count) still existed in the range between 3- and 4-log cells/ml. Significantly high numbers of HPC and DVC were found both in biofilm and in the effluent of the model system. The pipe material, exposure time, and the season were all relevant to the concentrations of VBNC and HPC bacteria detected. These findings indicate the importance of determining the number of VBNC cells and the type of pipe materials to estimate the HPC concentration in water distribution systems and thus the need of determining a DVC in evaluating disinfection efficiency.

• The Plant Pathology Journal
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• 제29권4호
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• pp.374-385
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• 2013

Environmental stresses induce several plant pathogenic bacteria into a viable but nonculturable (VBNC) state, but the basis for VBNC is largely uncharacterized. We investigated the physiology and morphology of the copper-induced VBNC state in the plant pathogen Ralstonia solanacearum in liquid microcosm. Supplementation of $200{\mu}M$ copper sulfate to the liquid microcosm completely suppressed bacterial colony formation on culture media; however, LIVE/DEAD BacLight bacterial viability staining showed that the bacterial cells maintained viability, and that the viable cells contain higher level of DNA. Based on electron microscopic observations, the bacterial cells in the VBNC state were unchanged in size, but heavily aggregated and surrounded by an unknown extracellular material. Cellular ribosome contents, however, were less, resulting in a reduction of the total RNA in VBNC cells. Proteome comparison and reverse transcription PCR analysis showed that the Dps protein production was up-regulated at the transcriptional level and that 2 catalases/peroxidases were present at lower level in VBNC cells. Cell aggregation and elevated levels of Dps protein are typical oxidative stress responses. $H_2O_2$ levels also increased in VBNC cells, which could result if catalase/peroxidase levels are reduced. Some of phenotypic changes in VBNC cells of R. solanacearum could be an oxidative stress response due to $H_2O_2$ accumulation. This report is the first of the distinct phenotypic changes in cells of R. solanacearum in the VBNC state.

• Food Science of Animal Resources
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• 제29권1호
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• pp.47-54
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• 2009

Predictive modeling was applied to study the growth of microorganisms related to spoilage in frankfurter sausage containing various levels of dietary fiber (0, 1, 2, and 3%) from rice bran and to estimate its shelf-life. Using the Baranyi model, total viable cells, anaerobic and psychrotrophic bacteria were measured during 35 days of cold storage ($<4{\pm}1^{\circ}C$). The lag times (LT) demonstrated by control and treatment groups were 6.28, 623, 6.24, and 6.25 days, respectively. The growth rate of total viable cells in each group were 0.95, 0.91, 0.92, and 0.91 (Log CFU/g/day), respectively. The anaerobic and psychrotrophic bacteria had lower initial ($y_0$) and maximal bacterial counts ($y_{max}$) than total viable cells. Also, the anaerobic and psychrotrophic bacteria possessed lower growth rate and longer lag time than total viable cells. The estimated shelf-life of frankfurter containing rice bran fiber by the growth rate of total viable cells was 7.8, 7.9, 7.9, and 7.7 days, respectively. There were no significant differences in shelf-life as a function of fiber content. In other words, the addition of dietary fiber in sausage did not show the critically hazardous results in growth of microorganism. The 12 predictive models were then characterized by high $R^2$, and small RMSE. Furthermore, $B_f$ and $A_f$ values showed a very close relationship between the predictive and observed data.

• Biotechnology and Bioprocess Engineering:BBE
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• 제3권2호
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• pp.51-54
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• 1998

It has been proved that co-cultivation of human neroblastoma cells and human fibroblast cells can enhance nerve cell growth and the production of BDNF in perfusion cultivation. In batch co-cultivation, maximum cell density was increased up to 1.76${\times}$106 viable cells/mL from 9${\times}$105 viable cells/mL of only neuroblastoma cell culture. The growth of neuroblastoma cells was greatly improved by culturing both nerve and fibroblast cells in a perfusion process, maintaining 1.5${\times}$106 viable cells/mL, which was much higher than that form fed-batch cultivation. The nerve cell growth was greatly enhance in both fed-batch and perfusion cultivations while the growth of fibroblast cells was not. It strongly implies that the factors secreted from human fibrobast cells and/or the environments of co-culture system can enhance both cell growth and BDNF secretion. Specific BDNF production rate was not enhanced in co-cultures; however, the production period was increased as the cell growth was lengthened in the co-culture case. Competitive growth between nerve cells and fibroblast cells was not observed in all cases, showing no changes of fibroblast cell growth and only enhancement of the neuroblastoma cell growth and overall BDNF production. It was also found that the perfusion cultivation was the most appropriate process for cultivating two cell lines simultaneously in a bioreactor.

• Korean Journal of Microbiology
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• 제52권3호
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• pp.313-318
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• 2016

Bacteria in the viable but nonculturable (VBNC) state fail to produce colonies on routine bacteriological media, but are still alive in the state of very low metabolic activity. The aim of the present study was to induce the VBNC state of the Edwardsiella tarda using sea water microcosm under starvation conditions at $10^{\circ}C$ and to investigate resuscitation of the VBNC cells in temperatures changed from 10 to $25^{\circ}C$, with and without additives. E. tarda entered into the VBNC state within about 42-84 days of incubation in the microcosm. Throughout this period, the total cell counts as determined using acridine orange direct counting remained near the original inoculum level of ${\sim}10^8cells/ml$. The live cell counts measured with direct viable counting, on the other hands, declined to ${\sim}10^4cells/ml$. When the VBNC cells were incubated with addition of yeast extract, fish muscle extract or serum at $25^{\circ}C$, the ratios of resuscitated samples were 37%, 23%, and 37%, respectively. The characteristics of resuscitated E. tarda were consistent with those of the original E. tarda. When the resuscitated E. tarda were intraperitoneally injected into olive flounders, all fishes died within 5 days, indicating that the VBNC E. tarda might retain its pathogenic potential. Therefore, E. tarda under starvation conditions in the winter enter into the VBNC state and the VBNC E. tarda cells resuscitated at summer and autumn seawater temperature are considered to be pathogen continuously to olive flounder on the southern coast of Korea.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.390-393
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• 2001

The optimal temperature, pH and aeration rate for spore production by Bacillus polyfermenticus SCD in 500 ml shake flask and 5-1 jar fermenter were found to be $32^{\circ}C$, 7.0 and 1.0 vvm. respectively. When batch culture processes was performed under optimized culture conditions. viable cells were $3.3{\times}10^{10}$ CFU/ml and spore cells were $3.3{\times}10^{10}$ CFU/ml. Fed-batch culture processes were also examined with regard to higer maximum viable cell and spore production. The highe viable cells and spores were obtained in 5-1 jar fermenter at 72 h cultivation time by strategy in an intermediate feeding mode with 60% glucose solution 150 ml and 5% soybean flour solution 150 ml fed to the fermenter twice, and the productivity of spore cells was significantly increased. Finally. volumetric productivity of spore cells on fed-batch culture indicated $9.9{\times}10^8$ CFU/ml/h, which was approximately 2 times higher than batch culture. Thus, fed-batch culture show a promise as an industrial production method.