• Title/Summary/Keyword: Bile culture

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Probiotic Potential of Pediococcus acidilactici and Enterococcus faecium Isolated from Indigenous Yogurt and Raw Goat Milk

  • Sarkar, Shovon Lal;Hossain, Md. Iqbal;Monika, Sharmin Akter;Sanyal, Santonu Kumar;Roy, Pravas Chandra;Hossain, Md. Anwar;Jahid, Iqbal Kabir
    • Microbiology and Biotechnology Letters
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    • v.48 no.3
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    • pp.276-286
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    • 2020
  • Probiotics are live microorganisms that, when administered in adequate amounts, confer health benefits to the host. This study was conducted for the isolation of potential lactic acid bacteria (LAB) with probiotic properties from goat milk and yogurt. Several tests were conducted in vitro using the standard procedures for evaluating the inhibitory spectra of LAB against pathogenic bacteria; tolerance to NaCl, bile salt, and phenol; hemolytic, milk coagulation, and bile salt hydrolase activities; gastrointestinal transit tolerance; adhesion properties; and antibiotic susceptibility. Among 40 LAB strains screened according to culture characteristics, five isolates exhibited antagonistic properties. Three were identified as Pediococcus acidilactici, and two were identified as Enterococcus faecium, exploiting 16S rRNA gene sequencing. All the isolates succeeded in the gastrointestinal transit tolerance assay and successively colonized mucosal epithelial cells. Based on the results of these in vitro assays, both P. acidilactici and E. faecium can be considered as potential probiotic candidates.

Probiotic Properties and Optimization of Gamma-Aminobutyric Acid Production by Lactiplantibacillus plantarum FBT215

  • Kim, Jaegon;Lee, Myung-Hyun;Kim, Min-Sun;Kim, Gyeong-Hwuii;Yoon, Sung-Sik
    • Journal of Microbiology and Biotechnology
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    • v.32 no.6
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    • pp.783-791
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    • 2022
  • Gamma-aminobutyric acid (GABA) improves various physiological illnesses, including diabetes, hypertension, depression, memory lapse, and insomnia in humans. Therefore, interest in the commercial production of GABA is steadily increasing. Lactic acid bacteria (LAB) have widely been reported as a GABA producer and are safe for human consumption. In this study, GABA-producing LAB were preliminarily identified and quantified via GABase assay. The acid and bile tolerance of the L. plantarum FBT215 strain were evaluated. The one-factor-at-a-time (OFAT) strategy was applied to determine the optimal conditions for GABA production using HPLC. Response surface methodology (RSM) with Box-Behnken design was used to predict the optimum GABA production. The strain FBT215 was shown to be acid and bile tolerant. The optimization of GABA production via the OFAT strategy resulted in an average GABA concentration of 1688.65 ± 14.29 ㎍/ml, while it was 1812.16 ± 23.16 ㎍/ml when RSM was applied. In conclusion, this study provides the optimum culture conditions for GABA production by the strain FBT215 and indicates that L. plantarum FBT215 is potentially promising for commercial functional probiotics with health claims.

Assessment of Characteristics and Functional Properties of Lactobacillus Species Isolated from Kimchi for Dairy Use

  • Baick, Seung-Chun;Kim, Cheol-Hyun
    • Food Science of Animal Resources
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    • v.35 no.3
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    • pp.339-349
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    • 2015
  • The objective of this study was to identify lactic acid bacteria (LAB) isolated from kimchi and to evaluate its characteristics and functional properties for application in fermented dairy products as a probiotic or commercial starter culture. Eight stains isolated from kimchi were selected through an investigation of phenotypic characteristics. Two strains (DK211 and DK303) were identified as Lactobacillus plantarum, another two (DK207 and DK215) as Lactobacillus paracasei, and one (DK301) as Lactobacillus sakei. The remaining three strains were identified as species of Weissella. All selected Lactobacillus strains had acid and bile tolerance, even though there was wide variation in the ability of each strain. DK303 showed a remarkably higher proteolytic activity. There were no significant differences in β-galactosidase activity among the tested strains, except that DK301 showed no activity. Auto-aggregation varied between 82.1 and 90.0%, and hydrophobicity values ranged from 0.5 to 51.6%.The strongest auto-aggregation and hydrophobicity were observed in DK211. All selected strains showed better 1,1-diphenyl-2-picrylhydrzyl (DPPH) scavenging activity than commercial strains. DK211, DK215, DK301, and DK303 had effective inhibitory activity against all pathogens tested except E. coli. When selected strains were used for yogurt preparation as a single starter culture, the time required to reach target titratable acidity (0.9) was 11-12 h. The yogurt fermented with DK211 had favorable panelists ratings for most sensory attributes, which were comparable with yogurt fermented with a commercial strain. The results suggest that strains isolated from kimchi could be potential probiotic and starter cultures for use in yogurt manufacturing.

Evaluation of Microencapsulated Local Isolates Lactobacillus casei 97/L3 and Lactobacillus delbrueckii 94/L4 for Improved Probiotic and Yogurt Starter Culture Application

  • Juvi, Denny;Sthefanie, Sthefanie;Sugata, Marcelia;Lucy, Jap;Andrian, Danish;Rizkinata, Denny;Michelle, Michelle;Jan, Tan Tjie
    • Microbiology and Biotechnology Letters
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    • v.47 no.2
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    • pp.211-219
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    • 2019
  • The effect of microencapsulation on previously isolated Lactobacillus delbrueckii 94/L4 as starter culture for yogurt, and Lactobacillus casei 97/L3 as a probiotic candidate was investigated. Preliminary results showed that L. delbrueckii 94/L4 exhibited tolerance to bile, unlike L. casei 97/L3. Freeze drying significantly (p < 0.05) reduced the viability of both isolates by log 0.71-2.70. Although microencapsulation preserved the viability of L. casei 97/L3 cells exposed to simulated gastrointestinal tract conditions for 120 min, it did not impart significant (p < 0.05) protection against loss of viability during the first 30 min of exposure. Conversely, microencapsulated L. delbrueckii 94/L4 with the addition of Streptococcus thermophilus 24/S1 as starter culture was successfully incorporated into milk to form yogurt, yielding a significantly (p < 0.05) improved product quality.

Effect of sodium polyanethol sulfonate on the isolation of Salmonella typhi from blood culture (Sodium polyanethol sulfonate의 혈액배양에서의 Salmonella typhi 분리에 대한 영향)

  • Chong, Yun-Sop
    • The Journal of the Korean Society for Microbiology
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    • v.9 no.1
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    • pp.13-18
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    • 1974
  • Blood is one of the most important clinical specimens for the isolation of bacteria. A rapid isolation and a high isolation rate of bacteria are very important in blood culture because bacteremic patients are mostly in grave condition. Various blood culture media which support growth of most fastidious bacteria are available commercially. However, growth of bacteria are frequently delayed because of antibacterial activity of blood. Sodium polyanethol sulfonate(Liquoid) has been reported to inactivate the antibacterial substance and disrupt phagocytic cells. The beneficial effect of SPS is well recognized in the isolation of gram-positive bacteria. However, the effect does not seem to be prominent for gram-negative bacilli isolation mainly due to the rapidity of their growth. It has been experienced with Sal. typhi that the growth is much slower than that of other gram-negative bacilli. For the rapid growth of the organism, use of bile broth has been recommended. Although Sal. typhi is the most frequently isolated organism at present, about one half of total isolates are other organisms and, in case bile broth is used, other media which support growth of these organisms should be used together. Fluid thioglycollate medium(FTM) which is always used in blood culture to isolate anaerobes is inferior to brain heart infusion(BHI) for the isolation of aerobes. This study was done to determine the effect of SPS on the isolation of Sal. typhi from blood. During the Sep. 1973 to Sep. 1974 study period, 2460 blood cultures were made from the Severance hospital patients: BHI and FTM sets 1431 specimens, BHI with SPS(0.05%) and FTM sets 396 specimens, BHI and FTM with SPS sets 359 specimens, BHI and BHI with SPS sets 274 specimens. Mean incubation time required for the macroscopic detection of growth of Sal. typhi were 3.5 days on BHI and 2.7 days on BHI with SPS. The 0.8 day difference was statistically significant. On FTM the mean incubation time was 3.8 days while it was 2.9 days on FTM with SPS. The 0.9 day difference was statistically significant. The result on BHI with and without SPS sets showed faster growth on BRI with SPS in 7 specimens and slower growth in one specimen and the remaining 12 showed growth at the same time. These specimens had mean incubation time of 3.2 days on BHI and 2.3 days on BHI with SPS. The 0.9 day difference was statistically significant. This study indicates beneficial effect of SPS for the rapid isolation of Sal. typhi from clinical blood specimens.

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Morphologic Changes in Microcystin-LR Treated Hepatocytes In vitro

  • Rhee, Seong-Hee;Kim, Bum-Seok;Lim, Chae-Woong
    • Toxicological Research
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    • v.22 no.3
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    • pp.301-306
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    • 2006
  • Microcystin-LR(MC-LR), a cyanobacterial toxin produced by Microcystis aeruginosa, causes severe hepatotoxicity. Here we investigated the morphologic changes of rat hepatocyte spheroid induced by exposure of MC-LR($10^{-6}M$) in vitro. In addition, to determine the effects of such toxin in the process of hepatocyte spheroid formation, primarily isolated hepatocytes were incubated with MC-LR and the process of spheroid formation was observed. In both hepatocyte spheroid and suspension culture systems, the morphologic changes caused by MC-LR were noticible at 5 min post exposure and were characterized by the loss of microvilli, cytoplasmic vacuolation, the accumulation of lipid droplets, and blob formation. Especially, the size and numbers of blob on the cell surface were increased as the incubation time prolonged and the appearance of electron dense bodies were observed in the cytoplasm of hepatocyte at 20 min post exposure. Furthermore, bile canaliculi-like structures in the hepatocyte spheroids were slightly widened and the process of spheroids formation was inhibited in the isolated hepatocytes incubated with MC-LR. These results indicate that morphologic changes in. the hepatocyte membrane and organelles seem to be typical events in showing the MC-LR induced hepatotoxic effects and the spheroid culture method might be a useful experimental tool to evaluate hepatoxicity since it reflects the in vivo status of hepatocytes.

Characterization of airag collected in Ulaanbaatar, Mongolia with emphasis on isolated lactic acid bacteria

  • Choi, Suk-Ho
    • Journal of Animal Science and Technology
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    • v.58 no.3
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    • pp.10.1-10.10
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    • 2016
  • Background: Airag, alcoholic sour-tasting beverage, has been traditionally prepared by Mongolian nomads who naturally ferment fresh mares' milk. Biochemical and microbiological compositions of airag samples collected in Ulaanbaatar, Mongolia and physiological characteristics of isolated lactic acid bacteria were investigated. Methods: Protein composition and biochemical composition were determined using sodium dodecyl sulfate-gel electrophoresis and high performance liquid chromatography, respectively. Lactic acid bacteria were identified based on nucleotide sequence of 16S rRNA gene. Carbohydrate fermentation, acid survival, bile resistance and acid production in skim milk culture were determined. Results: Equine whey proteins were present in airag samples more than caseins. The airag samples contained 0.10-3.36 % lactose, 1.44-2.33 % ethyl alcohol, 1.08-1.62 % lactic acid and 0.12-0.22 % acetic acid. Lactobacillus (L.) helveticus were major lactic acid bacteria consisting of 9 isolates among total 18 isolates of lactic acid bacteria. L. helveticus survived strongly in PBS, pH 3.0 but did not grow in MRS broth containing 0.1 % oxgall. A couple of L. helveticus isolates lowered pH of skim milk culture to less than 4.0 and produced acid up to more than 1.0 %. Conclusion: Highly variable biochemical compositions of the airag samples indicated inconsistent quality due to natural fermentation. Airag with low lactose content should be favorable for nutrition, considering that mares' milk with high lactose content has strong laxative effect. The isolates of L. helveticus which produced acid actively in skim milk culture might have a major role in production of airag.

Storage characteristics of frozen soy yogurt Prepared with different proteolytic enzymes and starter cultures (단백분해효소와 Starter Culture의 종류에 따른 frozen soy yogurt의 저장성)

  • Lee Sook-Young;Lee Jung-Eun
    • Korean journal of food and cookery science
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    • v.21 no.2 s.86
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    • pp.217-224
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    • 2005
  • The storage characteristics of frozen soy yogurt prepared with hydrolyzed soy protein isolates were evaluated. In order to facilitate lactic fermentation bacteria grow and produce lactic acid as fast rate as possible, soy protein isolate(SPI) was hydrolyzed using two kinds of proteases; bromelain and a-chymotrypsin. The cultural systems employed thereafter for lactic fermentations were Bifidobacterium bifidum or B. bifidum and Lactobacillus bulgaricus. The viable cell counts, normal- and bile acid tolerances from the mixed cultures of B. bifidum and L. bulgaricus decreased sharply during the initial first 3 days of frozen storage and then showed a gradual decrease thereafter. Melt-down percent of the all frozen products have been favorably affected as was shown by less melting at raised testing temperature during 28 days of frozen storage except for the initial 3 days during which a minor change has been observed. Among the various volatile flavor components, the contents of acetaldehyde, acetone, diacetyl and methanol generally increased during the frozen storage. In sensory test, the frozen soy yogurt prepared with a-chymotrypsin and mixed culture of B. bifidum and L. bulgaricus was the most desirable, the highest scores in sourness, bitterness and mouthfeel.

Role of Stem Cell Factor on the Recruitment of Mast Cells in the Development of Liver Fibrosis Induced by Bile Duct Ligation in the Rat (담관 결찰에 의한 간섬유증 발생에서 비만세포 동원에 미치는 Stem Cell Factor의 역할)

  • Jekal, Seung Joo;Ramm, Grant A.
    • Korean Journal of Clinical Laboratory Science
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    • v.36 no.2
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    • pp.163-172
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    • 2004
  • Mast cells (MCs) have been implicated in the pathogenesis of tissue fibrosis. However, the role of MC in the development of liver fibrosis has not been fully elucidated. Stem cell factor (SCF) is known to recruit MCs to the liver following injury as it induces mast cell proliferation, survival and differentiation from resident tissue precursors. This study examines the interaction between activated hepatic stellate cells (HSCs) and MCs in rat fibrotic liver, and SCF production by HSCs during culture in vitro. Rats were studied 4, 7, 14 and 21 days after bile duct ligation (BDL). Fibrogenesis was assessed by a measurement of collagen stained with sirius red F3B. Activated HSCs and MCs were identified by ${\alpha}$-smooth muscle actin (${\alpha}-SMA$) immunohistochemical and alcian blue staining and measured by a computerized image analysis system. SCF production was determined in rat HSC cultures using Western blotting. Mild fibrotic changes were noted in BDL rat livers as early as 4 days after induction of cholestasis. Significant expansion and organization of fibrous tissue has occurred in day 14 BDL rats which progressed to bridging fibrosis by day 21. In BDL rats, both a large number of activated HSCs and MCs were detected in portal tracts and fibrous septa. Both area of activated HSCs infiltration and density of MCs were significantly higher in all BDL group compared with Shams. In BDL rats, both areas of activated HSCs infiltration and density of MCs were no significant difference between day 4 and 7 and were significantly higher in day 14. However, the areas of activated HSCs infiltration were significantly lesser in day 21 and the densities of MCs were significantly higher in day 21 compared with day14 BDL. In BDL rats, both areas of activated HSCs infiltration and density of MCs were highly correlated with areas of fibrosis. Western blotting showed that SCF protein was consistently produced in activated HSCs by culture on plastic and freshly isolated HSCs expressed relatively little 30kD SCF compared to late primary culture activated HSCs (day 14) and passaged HSCs. These results suggest that HSCs activated in vitro produce SCF, and may play an important role in recruiting mast cells to the liver during injury and fibrosis.

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Identification and Characterization of Hydrogen Peroxide-generating Lactobacillus fermentum CS12-1

  • Kang, Dae-Kyung;Oh, H.K.;Ham, J.-S.;Kim, J.G.;Yoon, C.H.;Ahn, Y.T.;Kim, H.U.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.1
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    • pp.90-95
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    • 2005
  • Lactic acid bacteria were isolated from silage, which produce high level of hydrogen peroxide in cell culture supernatant. The 16S rDNA sequences of the isolate matched perfectly with that of Lactobacillus fermentum (99.9%), examined by a 16S rDNA gene sequence analysis and similarity search using the GenBank database, thus named L. fermentum CS12-1. L. fermentum CS12-1 showed resistance to low pH and bile acid. The production of hydrogen peroxide by L. fermentum CS12-1 was confirmed by catalase treatment and high-performance liquid chromatography. L. fermentum CS12-1 accumulated hydrogen peroxide in culture broth as cells grew, and the highest concentration of hydrogen peroxide reached 3.5 mM at the late stationary growth phase. The cell-free supernatant of L. fermentum CS12-1 both before and after neutralization inhibited the growth of enterotoxigenic Escherichia coli K88 that causes diarrhea in piglets.