• Title/Summary/Keyword: viable cell

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Improved Production of Live Cells of Lactobacillus rhamnosus by Continuous Cultivation using Glucose-yeast Extract Medium

  • Ling Liew Siew;Mohamad Rosfarizan;Rahim Raha Abdul;Wan Ho Yin;Ariff Arbakariya Bin
    • Journal of Microbiology
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    • v.44 no.4
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    • pp.439-446
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    • 2006
  • In this study, the growth kinetics of Lactobacillus rhamnosus and lactic acid production in continuous culture were assessed at a range of dilution rates $(0.05 h^{-1}\;to\;0.40h^{-1})$ using a 2L stirred tank fermenter with a working volume of 600ml. Unstructured models, predicated on the Monod and Luedeking-Piret equations, were employed to simulate the growth of the bacterium, glucose consumption, and lactic acid production at different dilution rates in continuous cultures. The maximum specific growth rate of L. rhamnosus, ${\mu}_{max}$, was estimated at $0.40h^{-1}$I, and the Monod cell growth saturation constant, Ks, at approximately 0.25g/L. Maximum cell viability $(1.3{\times}10^{10}CFU/ml)$ was achieved in the dilution rate range of $D=0.28h^{-1}\;to\;0.35h^{-1}$. Both maximum viable cell yield and productivity were achieved at $D=0.35h^{-1}$. The continuous cultivation of L. rhamnosus at $D=0.35h^{-1}$ resulted in substantial improvements in cell productivity, of 267% (viable cell count) that achieved via batch cultivation.

The Effect of Maesil(Prunus mime) Extract on the Acid Production and Growth of Yoghurt Starter (매실(Prunes mime) 착즙액이 Yoghurt Starter의 산생성 및 증식에 미치는 영향)

  • 박신인;이은희;남은숙
    • The Korean Journal of Food And Nutrition
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    • v.15 no.1
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    • pp.42-50
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    • 2002
  • This experiment was carried out to investigate the effect of Maesil extract on the acid production and growth of yoghurt starter in the skim milk medium. The Maesil extract was added to skim milk medium fur 1% to 9% and the medium was fermented by single or mixed culture of 4 types of lactic acid bacteria(Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus, bulgaricus, Lactobacillus casei). The chemical composition of Maesil, the changes in acid production (titratable acidity, pH) and number of viable cells of the medium during lactic fermentation in skim milk added with Maesil extract, and the keeping quality of curd yoghurts containing Maesil extract have determined. The composition of Maesil were 0.4% crude ash, 4.1% dietary fiber, 4.66%l citric acid, 0.264% total sugars and 405.34mg% vitamin C. The addition of Maesil extract stimulated the acid production and propagation of the lactic acid bacteria. Among the treatments tested, the addition of 3% Maesil extract with the mixed culture of Streptococcus thermophilus, Lactobacillus acidophilus and Lactobacillus casei produced the highest amount of acid(1.23%) and showed the highest number of viable cell counts(3.6$\times$10$^{11}$ cfu/mL). When the curd yoghurts containing 3% Maesil extract with the mixed culture of the lactic acid bacteria were kept at 4$^{\circ}C$ and 2$0^{\circ}C$ for 30 days, it was showed that the changes of titratable acidity, pH and number of viable cell counts of the lactic acid bacteria were not significantly different during storage. Therefore the keeping quality of the curd yoghurts adding 3% Maesil extract showed relatively good at the shelf-life.

Effect of Chlorella Extract on Acid Production and Growth of Yoghurt Starter (Chlorella 추출물 첨가가 Yoghurt Starter의 산 생성 및 증식에 미치는 영향)

  • 조은정;남은숙;박신인
    • The Korean Journal of Food And Nutrition
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    • v.17 no.1
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    • pp.8-17
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    • 2004
  • The effect of chlorella extract on the growth and acid production of yoghurt starter was investigated in order to prepare the yoghurt added with chlorella extract. The various levels of chlorella extract powder were added to skim milk medium and the medium was fermented by single or mixed culture of 4 types of lactic acid bacteria such as Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus casei, and Lactobacillus bulgaricus. The changes in acid production(pH, titratable acidity) and number of viable cells of the medium during fermentation in skim milk added with chlorella extract powder have determined. When chlorella extract powder was added to skim milk medium at the levels of 0.5%, 1.0%, 2.0%, and 3.0%, the addition of 0.5% chlorella extract powder with the single culture of Str. thermophilus, Lac. casei, and Lac. bulgaricus showed the highest number of viable cell counts after 9 hours incubation. And also all single cultures of the yoghurt starter produced the higher amounts of acid with the addition of 0.5% chlorella extract powder. When chlorella extract powder was added to the medium at the levels of 0.25%, 0.5%, 1.0%, and 2.0%, the addition of lower lever(0.25∼0.5%) of chlorella extract powder with the mixed culture of the lactic acid bacteria showed more the acidity of pH and the number of viable cell counts. Among the treatments tested, the addition of 0.25% chlorella extract powder with the mixed culture of Str. thermophilus and Lac. casei produced the highest number of viable cell counts after 12 hours incubation. Therefore it was suggested to manufacture the yoghurt with the addition of 0.25% chlorella extract powder and the inoculation of mixed culture of Str. thermophilus and Lac. casei for on the stimulation of growth of the yoghurt starter.

Comparative Evaluation of Probiotic Activities of Bifidobacterium longum MK-G7 with Commercial Bifidobacteria Strains

  • Jung, Hoo-Kil;Kim, Eung-Ryool;Ji, Geun-Eog;Park, Jong-Hyun;Cha, Seong-Kwan;Juhn, Suk-Lak
    • Journal of Microbiology and Biotechnology
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    • v.10 no.2
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    • pp.147-153
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    • 2000
  • This study was conducted to compare probiotic activities and physiological functions of Bifidobacterium longum Mk-G7 with weveral commercial and type strains of bifidobacteria. bif. longum MK-G7 showed the highest acid tolerance against HCl and acetic acid, whereas bif. infantis Y-1 showed the lowest acid tolerance and more than 4 log cycles of viable cell count decreased due to acid injuty. Viable cell counts of bifidobacteria strains decreased more than 1.5 log cycles owing to oxygen toxicity, with the exception of Bif. longum MK-G7, Bif. infantis Y-2, Bif. longum Y-3, Bif. longum Y-6, and Bif. longum RD-13 showed the highest bile tolerance, whereas Bif. longum MK-G7 showed a medium level of bile tolerance. Only Bif. longum MK-G7 howed much higher antibiotic resistance against both tetracycline and penicillin-G in the MIC(minimum inhibitory concentration) level of 24.8 mg/I and 0.52mg/I, respectively. Bif longum Y-6, and Bif. bifidum ATCC 29539 showed more than 80% of anti-mutagenicity against NQO(4-nitroquinolinel-oxide). Since the production of cytokines such as $TNF(tumor necrosis factor)-{\alpha}$ and IL (interleukin)-6, and NO(nitric oxide) in the macrophage cell line Raw 264.7 cells increased as Bif. longum MK-G7 cell concentration increased, ti was suggested that Bif. longum MK-G7 is able to enhance immunopotentiating activity in vitro. When freeze-dred Bif. longum MK-G7 was administered to mice at the dose of 1,2,4, and 6 g/kg of body weight, all of the mice survived in all feeding groups, proving the GRAS(generally recognized as safe) status of Bif. longum MK-G7. When fermented milk containing Bif. longum MK-G7 was administered to human volunteers, viable cell count of total bifidobacteria and anaerobes in the feces increased up to 0.5 log cycles more than before the administration. In particular, Bif. logum MK-G7 ingibited the growth of Bacteroides at the level of 1.0-1.5 log cycles.

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Adhesion of Kimchi Lactobacillus Strains to Caco-2 Cell Membrane and Sequestration of Aflatoxin B1 (김치 유산균의 Caco-2 세포막 부착성 및 Aflatoxin B1 제거 효과)

  • Lee, Jeongmin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.5
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    • pp.581-585
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    • 2005
  • Five lactic acid bacteria (LAB) including 2 Lactobacillus strains isolated from Kimchi were evaluated to determine the binding ability to Caco-2 cells and $AFB_1$. LAB were divided into three different groups ; viable, heat-treated, and acid-treated cells. In the radioactive-labeling assay for bound cell counting, viable Lactobacillus Plantarum KCTC 3099 showed the higher adhesion to Caco-2 cells with the binding capacity of $39.2\%$, which was $149\%$ higher than Lactobacillus rhamnosus GG as a positive control. Leuconostoc mesenteroids KCTC 3100 showed the similar binding ability to L. rhamnosus GG. After 1 hour incubation at $37^{\circ}C$ with $AFB_1$, viable L. Planterum KTCC 3099 removed the toxin by $49.8\%$, which was similar level to L. rhamnosus GG. Both heat- and acid-treated groups showed high binding effect but acid-treated group was more effective for both Caco-2 cell binding and $AFB_1$ removal than the other. These results indicate that components of bacterial cell wall might be involved in tile binding to intestinal cells and toxins.

Effects of Knockout Serum Replacement in the Culture Medium on the Proliferation of Porcine Fetal Fibroblasts In Vitro

  • Kim, Eun-Ju;Park, Jung-Joo;Choi, Young-Ju;Park, Sang Kyu;Roh, Sang-Ho
    • International Journal of Oral Biology
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    • v.35 no.1
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    • pp.1-5
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    • 2010
  • Human fibroblasts that maintain the structural integrity of connective tissues by secreting precursors of the extracellular matrix are typically cultured with serum. However, there are potential disadvantages of the use of serum including unnatural interactions between the cells and the potential for exposure to animal pathogens. To prevent the possible influence of serum on fibroblast cultures, we devised a serum-free growth method and present in vitro data that demonstrate its suitability for growing porcine fetal fibroblasts. These cells were grown under four different culture conditions: no serum (negative control), 10% fetal bovine serum (FBS, positive control), 10% knockout serum replacement (KSR) and 20% KSR in the medium. The proliferation rates and viabilities of the cells were investigated by counting the number of cells and trypan blue staining, respectively. The 10% FBS group showed the largest increase in the total number of cells ($1.09\;{\times}\;10^5\;cells/ml$). In terms of the rate of viable cells, the results from the KSR supplementation groups (20% KSR:64.7%; 10% KSR: 80.6%) were similar to those from the 10% FBS group (68.5%). Moreover, supplementation with either 10% ($3.0\;{\times}\;10^4\;cells/ml$) or 20% KSR ($4.8\;{\times}\;10^4\;cells/ml$) produced similar cell growth rates. In conclusion, although KSR supplementation produces a lower cell proliferation rate than FBS, this growth condition is more effective for obtaining an appropriate number of viable porcine fetal fibroblasts in culture. Using KSR in fibroblast culture medium is thus a viable alternative to FBS.

RT-PCR Targeting rpoB mRNA for Drug Susceptibility Test of Mycobacterium tuberculosis in Liquid Culture

  • Jin, Hyunwoo
    • Biomedical Science Letters
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    • v.22 no.4
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    • pp.215-219
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    • 2016
  • The problems of tuberculosis and its drug resistance are very severe. Therefore, rapid and accurate drug susceptibility assay is required. Recently, there has been an increased understanding of the genetic mechanism of Mycobacterium tuberculosis (MTB) drug resistance as well as advancement of molecular technologies. While many gene mutations correlate well with drug resistance, many genes do not show a strong correlation with drug resistance. For this reason, the current study assessed the utility of rpoB mRNA as a target to detect live mycobacteria. In this study, RT-PCR targeting of rpoB mRNA in BCG treated with rifampin was performed. Conventional RT-PCR and real-time PCR targeting rpoB mRNA as well as 85B mRNA was performed to determine whether these two methods could distinguish between viable and non-viable MTB. The levels of rpoB and 85B mRNA detected by RT- PCR were compared in parallel with colony forming unit counts of BCG that were treated with rifampin for different periods of time. The data suggests that that even though both mRNA levels of rpoB and 85B decreased gradually when rifampin-treatment increased, the rpoB mRNA seemed to represent live bacteria better than 85B mRNA. This study clearly indicates that RT-PCR is a good method to monitor viable cell counts in the liquid culture treated with the anti-tuberculosis drug.

Potential Health-Promoting Benefits of Paraprobiotics, Inactivated Probiotic Cells

  • Akter, Shahina;Park, Jong-Hyun;Jung, Hoo Kil
    • Journal of Microbiology and Biotechnology
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    • v.30 no.4
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    • pp.477-481
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    • 2020
  • Viability plays an important role in the beneficial microbes (probiotics) to produce health benefits. However, this idea has been changed after the invention of the term "paraprobiotics," indicating that non-viable microbes could produce health benefits similar to those produced by live probiotics. Occasionally, it might be dangerous to administer live probiotics to people with weak immunity. In such cases, ingestion of paraprobiotics could be a potential alternative. The definition of paraprobiotics refers to the use of inactivated (non-viable) microbial cells or cell fractions to provide health benefits to the consumer. Paraprobiotics have attracted much attention because of their long shelf life, safety, and beneficial effects, such as modulation of immunity, modification of biological responses, reduction of cholesterol, anti-inflammatory, and antiproliferative properties. These features indicate that paraprobiotics may play a vital role in improving the health of the consumer by enhancing particular physiological functions, even though the exact underlying mechanisms have not yet been completely elucidated. In this mini-review, we briefly discuss the historical backgrounds of paraprobiotics and evidence of their health-promoting effects, prophylactic, and therapeutic properties.

Production of Fermented Kale Juices with Lactobacillus Strains and Nutritional Composition

  • Kim, Seong Yeong
    • Preventive Nutrition and Food Science
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    • v.22 no.3
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    • pp.231-236
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    • 2017
  • Fermented kale juices using four types of lactobacilli were produced in the present study. After 48 h of fermentation time, viable cell counts of all ferments reached an above $10^9CFU/mL$. The viability of the ferments after cold storage in the refrigerator for 4 weeks showed $10^8CFU/mL$ in all ferments. Among four types of fermented kale juices, the ferment of Lactobacillus acidophilus IFO 3025 indicated a good nutritional composition, including neutral sugar ($1,909.76{\mu}g/mL$), reducing sugar ($564.00{\mu}g/mL$, P<0.05), and protein contents ($160.06{\mu}g/mL$, P<0.05). The results of mineral composition analysis had the highest potassium value in all ferments ($854.16{\sim}895.07{\mu}g/mL$), particularly in the ferment of Lactobacillus brevis FSB-1 (P<0.001), which is necessary to sustain osmotic pressure, prevention of high blood pressure, and protein synthesis. Moreover, calcium, phosphorus, and magnesium contents related to bone health were generally sufficient in all ferments. Consequently, in this study, fermented kale juices may be suggested as a healthy fermented beverage with essential nutrients. However, the acceptability of the fermented kale juice to the Korean taste should be further investigated with a trained taste panel to determine whether inoculated fermentation could be an option for the consumers.