• Title/Summary/Keyword: Culture Supernatant

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Antibacterial Activity of Lactobacillus casei Isolated from a Fermented Milk Against Pathogenic Enteric Bacteria (발효유(醱酵乳)에서 분리된 Lactobacillus casei의 병원성 장내세균에 대한 항균작용)

  • Choi, Chul-Soon;Chung, Jae-Bong;Chung, Sang-In;Yang, Yong-Tae
    • The Journal of the Korean Society for Microbiology
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    • v.19 no.1
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    • pp.41-48
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    • 1984
  • The present study was undertaken to elucidate the antibacterial spectrum of L. casei phage type $J_1$ strain isolated from a fermented milk product against pathogenic enteric bacteria. Growth inhibitory effects and minimum inhibitory concentration(MIC) of culture supernatants of L. casei grown in MRS broth were measured by both plate culture method and microplate broth dilution technique against Salmonella typhi, Salmonella typhimurium, Shigella flexneri, Shigella dysenteriae, enterpathogenic E. coli, Klebsiella pneumoniae and Pseudomonas aeruginosa. The results are summarized as follows: 1. The MRS broth culture of L. casei gave a similar extent of growth inhibitory effects against S. typhi, S. typhimurium, S. flexneri, S. dysenteriae, E. coli, K. pneumoniae and P. aeruginosa, respectively. 2. The inhibitory effects of L. casei culture were observed either in whole broth culture or in culture supernatant, but neither the bacterial suspension nor the neutralized culture supernatant showed such as antibacterial activities. 3. The MIC titres of the culture supernatants were ${\log_2}5$ to ${\log_2}6$, whereas those of the neutralized culture supernatant dropped markdely to ${\log_2}2$ to ${\log_2}3$. These results indicated that major portion of growth inhibitory effects of MRS broth culture of L. casei against enteric bacterial pathogens was possibly due to the acids produced, and minor portion to other antibacterial substances.

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Antibacterial Activity against Pathogenic Bacteria of Lactiplantibacillus argentratensis Isolated from Blueberries (블루베리로부터 분리된 Lactiplantibacillus argentratensis의 병원성균에 대한 항균활성)

  • Natsag Lkhagvasuren;Gil-Ha Kim;Batchimeg Namshir;Woan Sub Kim
    • Journal of Dairy Science and Biotechnology
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    • v.41 no.4
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    • pp.191-202
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    • 2023
  • In this study, lactic acid bacteria (LAB) was isolated from blueberries. The isolated LAB were rod-shaped and gram-positive, as shown using gram staining. In addition, the identified bacteria showed high homology to Lactiplantibacillus argentoratensis. The culture supernatant was isolated from L. argentoratensis and its antibacterial activity against the pathogenic bacteria Salmonella and Escherichia was analyzed. Culture supernatants of L. argentoratensis significantly inhibited the growth of Salmonella. Enteritidis NCCP 16947, Salmonella Typhimurium NCCP 16960, and Salmonella. Thompson NCCP 11704. Interestingly, the higher the concentration of the culture supernatant, the more significant was the antibacterial activity. Additionally, the culture supernatant of L. argentoratensis showed significant antibacterial activity against Escherichia strains. To determine whether the antibacterial substance is stable to heat and pH, the LAB culture supernatant was heat-treated under 65℃ for 30 min, 75℃ for 15 min, 85℃ for 10 min, and 100℃ for 5 min. Measurement of antibacterial activity against pathogenic strains by adding 5% of heat-treated culture medium showed the same antibacterial activity as before heat treatment. However, in a test where the pH of the culture supernatant was adjusted to 7.0 from 3.73, no antibacterial activity was observed.

Antioxidative and Cholesterol-reducing Activity of Bacillus polyfermenticus SCD (Bacillus polyfermenticus SCD의 항산화 및 콜레스테롤 저하효과)

  • 정황영;김태훈;박준석;김기태;백현동
    • KSBB Journal
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    • v.18 no.5
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    • pp.371-376
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    • 2003
  • Antioxidative and cholesterol-reducing activity of Bacillus polyfermenticus SCD were measured to characterize its probiotic properties. DPPH (1,1-diphenyl-2-picyryl hydrazyl) radical scavenging activity of the culture supernatant of B. polyfermenticus SCD was estimated to be 48%. The culture supernatant on the peroxidation of linoleic acid were investigated and the value was shown to be about 45%. The inhibition of TBA (2-thiobarbituric acid) formation of the culture supernatant was revealed 60% when stimulator was presented. The SOD-like activity of the culture supernatant was about 15%, which is similar to BHT (butylated hydroxytoluene) and ${\alpha}$-tocopherol. After cultured in TSB broth added soluble cholesterol either 0.1% or 0.3% of oxgall in 37$^{\circ}C$ for 24 h aerobically, cholesterol-reducing activities were revealed about 67% or 64%, respectively. To test whether the products are cholesterol-related or not, residual activity was determined. The cholesterol activity was rarely changed. In addition, when the cell extracts recovered after cultivation, was tested in absence of cholesterol, cholesterol activity was not detected. However, cholesterol activity was detected in the presence of cholesterol. Thus, it was assumed that B. polyfermenticus SCD could reduce cholesterol by conjugating with it, rather than by digesting the cholesterol using cholesterol-hydrolyzing enzymes.

Secretion of Cytokine Stimulating Intercellular Adhesion Molecule-l Expression from THP-l Cells Infected with Human Cytomegalovirus (HCMV에 감염된 TBP-1 세포에서 세포간 부착분자-1 발현 촉진물질의 방출)

  • Kim, Mi-Suk;Yi, Hyun-Ah;Lee, Chan-Hee
    • Korean Journal of Microbiology
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    • v.44 no.2
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    • pp.140-146
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    • 2008
  • Human cytomegalovirus (HCMV) stimulates the expression of intercellular adhesion molecule (ICAM-l) on the surface of monocytic THP-1 cells. Stimulation of ICAM-l did not require HCMV gene expression since UV-inactivated HCMV (UV-HCMV) was able to induce ICAM-l expression. ICAM-l expression was also stimulated in uninfected THP-l cells which were fed with culture supernatant of HCMV-infected THP-l cells. Co-culture experiment using trans-well insert supported that HCMV-infected THP-l cells secreted some cytokine(s) stimulating ICAM-l expression. The stimulation of ICAM-l by HCMV-infected cell culture supernatant appears to involve $NF-{\kappa}B$ pathway. Culture supernatant from THP-l cells infected with UV-HCMV, whose gene expression was abrogated, failed to stimulate ICAM-l expression on naive THP-l cells. Thus, HCMV gene expression seems to be required in secretion of cytokine(s) stimulating ICAM-l expression.

Cytokines Stimulate Lung Epithelial Cells to Release Nitric Oxide

  • Robbins, Richard A.;Kwon, O-Jung
    • Tuberculosis and Respiratory Diseases
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    • v.42 no.4
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    • pp.447-454
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    • 1995
  • Cytokine release from alveolar macrophages and subsequent interaction of these cytokines with the bronchial epithelium can induce epithelial cells to release inflammatory mediators. Nitric oxide(NO), a highly reactive gas formed from arginine by nitric oxide synthase(NOS), is known to be involved in inflammation and edema formation, and the inducible form of NOS(iNOS) can be increased by cytokines. In this context, we hypothesized that lung epithelial cells could be stimulated by cytokines released by alveolar macrophages to express iNOS. To test this hypothesis, the murine lung epithelial cell line, LA-4, or the human lung epithelial cell line, A549, were stimulated with culture supernatant fluids from alveolar macrophages. NO production was assessed by evaluating the culture supernatant fluids for nitrite and nitrate, the stable end products of NO. Both murine and human cell culture supernatant fluids demonstrated an increase in nitrite and nitrate which were time- and dose-dependent and attenuated by $TNF{\alpha}$ and IL-$1{\beta}$ antibodies(p<0.05, all comparisons). Consistent with these observations, cytomix a combination of $TNF{\alpha}$, IL-$1{\beta}$, and $\gamma$-interferon, stimulated the lung epithelial cell lines as well as primary cultures of human bronchial epithelial cells to increase their NO production as evidenced by an increase in nitrite and nitrate in their culture supernatant fluids, an increase in the iNOS staining by immunocytochemistry, and an increase in iNOS mRNA by Northern blottin(p<0.05, all comparisons). The cytokine effects on iNOS were all attenuated by dexamethasone. To determine if these in vitro observations are reflected in vivo, exhaled NO was measured and found to be increased in asthmatics not receiving corticosteroids. These data demonstrate that alveolar macrophage derived cytokines increase iNOS expression in lung epithelial cells and that these in vitro observations are mirrored by increased exhaled NO levels in asthmatics. Increased NO in the lung may contribute to edema formation and airway narrowing.

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Inhibition of Aflatoxin Production of Aspergillus flavus by Lactobacillus casei

  • Chang, In-Jeong;Kim, Jeong-Dong
    • Mycobiology
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    • v.35 no.2
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    • pp.76-81
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    • 2007
  • Lactobacillus casei KC-324 was tested for its ability to inhibit aflatoxin production and mycelial growth of Aspergillus flavus ATCC 15517 in liquid culture. flatoxin $B_{1}$ biosynthesis and mycelial growth were inhibited in both simultaneous culture and individual antagonism assays, suggesting that the inhibitory activity was due to extracellular metabolites. produced in cell-free supernatant fluids of the cultured broth of L. casei KC-324. In cell-free supernatant fluids of all media tested, deMan, Rogosa and Sharpe broth, potato dextrose broth, and Czapek-Dox broth+1% yeast extract showed higher antiaflatoxigenic activity. In these case, fungal growths, however, was not affected as measured by mycelial dry weight. The antiaflatoxigenic metabolites from L. casei KC-324 were produced over wide range of temperatures between $25^{\circ}C$ and $37^{\circ}C$. However, these metabolites were not thermostable since the inhibitory activity of the supernatant was inactivated within 30 minutes at $100^{\circ}C$ and $121^{\circ}C$. The inhibitory activity was not influenced by changing pH of supernatant between 4 and 10. However, the antiaflatoxigenic activity was slightly reduced at pH 10.

Control Effect of Dinoflagellate Bloom by Powder of Marine Rock and Fungus Culture Supernatant (해양암석 분말과 곰팡이 배양액에 의한 적조생물 편조류의 구제효과)

  • Hyun, Sung-Hee;Shin, Hyun-Woung
    • Korean Journal of Microbiology
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    • v.42 no.1
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    • pp.7-11
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    • 2006
  • To see effect of marine rock powder and fungal culture supernatant, we analyzed the biodegradation rates of harmful marine dinoflagellate, Heterosigma akashiwo and Prorocentrum minimum for developing the effective control methodology of algal bloom. Relatively low removal rates were observed in the treatment of marine rock powder or buffer solution alone. However, the lysis of H. akashiwo and P. minimum was enhanced in the combined treatments of marine rock powder with fungal supernatant. The effective concentration and exposure time of fungal supernatant for the lysis of H. akashiwo and P. minimum were 5 ml/l and 30 minutes, respectively. These results suggest that the fungal supernatant may be a biocontrol agent for the control of algal blooms in seawater.

FACTORS AFFECTING THE FORMATION OF SOLUBLE GLUCAN BY LACTOCOCCUS LACTIS 1370 (유산구균 1370의 수용성 글루캔 형성에 영향을 미치는 인자)

  • Oh, In-Gyun;Yang, Kyu-Ho;Chung, Jin;Oh, Jong-Suk
    • Journal of the korean academy of Pediatric Dentistry
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    • v.27 no.2
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    • pp.185-191
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    • 2000
  • The water-soluble glucan produced by Lactococcus lactis 1370 affects the formation of dental plaque by Streptococcus mutans. In this study the factors affecting the formation of water-soluble glucan were assessed as the optical density of culture supernatant of Lactococcus lactis 1370 in the spectrophotometer. 1. The optical density of culture supernatant was high when Lactococcus lactis 1370 was cultured in M17 broth with 5% sucrose, while being low in culture supernatant of Streptococcus mutans. 2. The optical density of culture supernatant was higher when Lactococcus lactis 1370 was cultured in M17 broth with 10% sucrose than when being cultured without sucrose (p<0.05), and was higher at pH 7 than pH 5 (p<0.05). 3. The optical density of culture supernatant was the highest at $37^{\circ}C$ among $32^{\circ}C,\;37^{\circ}C\;and\;42^{\circ}C$, and was higher in the anaerobic incubator than in the aerobic incubator (p<0.05). 4. The optical density of culture supernatant was the highest in the media containing 1.0mM $CaCl_2$ (p<0.05), 2.5mM KCl (p<0.05), and 1.6mM $MgCl_2$. 5. When Streptococcus mutans was cultured in the media containing a quarter culture supernatant of Lactococcus lactis 1370 grown in M17 broth, the mean weight of produced artificial plaque was 103.0mg on the wire, whereas being significantly reduced to 5.6mg in the media containing a quarter culture supernatant of Lactococcus lactis 1370 grown in M17 broth containing 5% sucrose (p<0.05). These results indicate that the water-soluble glucan is more formed by Lactococcus lactis 1370 in the media containing sucrose or under the adequate conditions for the growth of bacteria, and inhibits the formation of artificial plaque by Streptococcus mutans.

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Isolation of lysozyme producing bacteria capable of solubilizing microbial cells (미생물 용해가 가능한 Iysozyme 분비 균주의 분리 및 특성)

  • Guo, Pengfei;Seo, Sun-Keun;Zhang, Lei;Kim, Hyo-Sang;Oh, Young-Khee;Jahng, Deok-Jin
    • KSBB Journal
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    • v.23 no.3
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    • pp.187-192
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    • 2008
  • Lysozyme-producing microorganisms were isolated to obtain bacteria which can efficiently solubilize microbial cells. Cells of normal and chloroform-treated Escherichia coli and Micrococcus Iysodeikticus were used as model substrates to isolate lysozyme-producing microorganisms and investigate the efficiency of cell lysis. The culture supernatant of the isolate New1 (98% similarity of 16S rDNA sequence with Thermomonas haemolytica) showed different lytic characteristics for different substrates. Thermal treatment (autoclave) of substrate cells showed a significant effect on cell solubilization by culture supernatant of the New1. For autoclaved substrate cells, E. coli, M. Iysodeikticus and chloroform-treated E. coli were solubilized by 58.7%, 49.4% and 79.1%, respectively, in the culture supernatant of New1. The lytic activity of New1 was mainly caused by lysozyme produced by the isolate. It was also showed that New1 exhibited high protease activity and a little cellulase activity.

Immunoenhancing Effect of 1,2-Benzopyrone on the Oxidative Burst Activity to Phagocytic Response of Canine Peripheral Blood Phagocytes (개 말초혈액 탐식세포의 탐식반응에 따른 순간산소 과소비현상에 있어 1,2-benzopyrone의 면역증강효과)

  • 김현아;강지훈;양만표
    • Journal of Veterinary Clinics
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    • v.21 no.3
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    • pp.236-242
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    • 2004
  • 1,2-benzopyrone can stimulate macrophages to increase the ability of phagocytosis. Peripheral blood polymorphonuclear cells (PMN) and macrophages destroy microbial organisms with reactive oxygen species (ROS), called oxidative burst activity (OBA). This study was undertaken to determine whether 1,2-benzopyrone affects the OBA on the phagocytic response of canine peripheral blood phagocytes. The OBA of phagocytes in the addition or absence of latex beads was analyzed by flow cytometry system using dihydrorhodamine 123 (DHR). The direct treatments of 1,2-benzopyrone have no effect on the OBA of peripheral blood mononuclear cells (PBMC), PMN and monocyte-rich cells regardless of addition of latex beads. When latex beads are added to PMN, the OBA of PMN was remarkably enhanced by culture supernatant from PBMC but not PMN treated with 1,2-benzopyrone. Similary, it was also enhanced by human recombinant (hr) $TNF-\alpha.$ However, when latex beads were not added to PMN, its OBA was not enhanced by culture supernatant from either PBMC or PMN treated with 1,2-benzopyrone. The OBA of latex beads-phagocytized PBMC and monocyte-rich cells was not enhanced by culture supernatant from either PBMC or PMN treated with 1,2-benzopyrone. These results strongly suggested that 1,2-benzopyrone has an immunoenhancing effect on the OBA of PMN when phagocytic response occurred only. This enhanced OBA may be mediated through active humoral substance(s), such as $TNF-\alpha,$ produced by PBMC stimulated with 1,2-benzopyrone.