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

• 미생물학회지
• /
• 제42권3호
• /
• pp.205-209
• /
• 2006

호소수내의 '살아있는 세균'을 측정하기 위하여 quantitative direct viable count (qDVC) 방법을 적용하였다. qDVC방법에 적용되는 최적 glycine 농도는 2%였으며, '살아있는 세균'을 계수하는데 있어서 평판계수법, CTC법 보다는 qDVC 방법이 보다 효과적이라는 것을 확인하였다. qDVC방법으로 '살아있는 세균'을 측정한 결과 다른 두 방법보다 $2.4{\sim}6.0$배 높은 값이었다. 또한 qDVC방법은 '살아있는 세균'을 죽은 세포 또는 휴면세포와 쉽게 구별할 수 있었다.

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

• 한국수산과학회지
• /
• 제29권6호
• /
• pp.882-887
• /
• 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
• /
• 제17권9호
• /
• pp.1558-1562
• /
• 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
• /
• 제29권4호
• /
• pp.374-385
• /
• 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.

• 한국축산식품학회지
• /
• 제29권1호
• /
• pp.47-54
• /
• 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
• /
• 제3권2호
• /
• pp.51-54
• /
• 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.

• 미생물학회지
• /
• 제52권3호
• /
• pp.313-318
• /
• 2016

Viable but nonculturable (VBNC) 상태에 들어간 세균은 일반적인 증균 배지에서는 집락을 형성하지 않지만, 죽은 것이 아니라 낮은 대사활성상태로 유지되고 있다. 본 연구에서는 $10^{\circ}C$의 저온 빈영양 해수에서 Edwardsiella tarda를 VBNC 상태로 유도한 후, 해수 온도를 10에서 $25^{\circ}C$로 상승시킬 때 첨가된 유기물의 종류에 따라 VBNC 상태인 균의 소생 가능성을 알아보고자 하였다. E. tarda가 접종된 빈영양 해수 microcosm을 $10^{\circ}C$에 유지하였을 때 VBNC 유도 기간은 42-84일까지 다양하였다. 유도 기간 동안 acridine orange direct counting법으로 계수한 총 균수는 초기 접종 농도인 약 $10^8cells/ml$로 일정하였으며, direct viable counting법으로 계수한 생존 균수는 약 $10^4cells/ml$로 감소되었다. VBNC E. tarda에 효모추출물, 넙치근육추출물 그리고 혈청을 첨가하여 $25^{\circ}C$에서 소생을 유도한 결과 전체 시료 개수의 37%, 23%, 37%에서 각각 소생이 확인되었으며 소생된 E. tarda의 특성은 VBNC 유도 전 원래의 세균과 일치하였다. 소생된 E. tarda를 넙치(Paralichthys olivaceus)에 복강 주사 하였을 때 접종 후 5일 이내에 시험어가 모두 사망함으로써 VBNC 상태의 E. tarda가 독력을 유지하고 있었음을 시사하였다. 그러므로 E. tarda는 우리나라 남해 연안 겨울의 저온 빈영양 해수에서 VBNC 상태로 유도되었다가 여름과 가을 시기에 수온 상승과 더불어 소생되어 양식 넙치에 지속적인 발병 요인이 되고 있는 것으로 생각된다.

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