• Journal of Dairy Science and Biotechnology
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• v.37 no.1
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• pp.40-48
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• 2019

Bifidobacteria are a prototype probiotic, which normally inhabit the intestinal tract of humans. In the present study, four species of Bifidobacterium isolated from the feces of infants were characterized. The tolerance for acid or bile salt, autoaggregation, and antibiotic resistance of the bacteria were examined. The four species were resistant to low pH, bile salts, and up to 3% bile acid. Autoaggregation rates were as high as 90%. The bacteria were consistently resistant to gentamicin, kanamycin, streptomycin, ciprofloxacin, and nalidixic acid. Due to their tolerance to environmental factors like acid and bile salts, B. longum MG723, B. breve MG729, B. bifidum MG731, and B. animalis subsp. lactis MG741 are potentially valuable as probiotics and may be useful for industrial application.

• Korean Journal of Microbiology
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• v.51 no.3
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• pp.241-248
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• 2015

Although studies on probiotics have been performed mostly with viable microbes, the beneficial functions of dead or heat-killed form of probiotic strains have also been examined. In this study, live and heat-killed forms of Lactobacillus acidophilus CBT LA1 were investigated in vitro and in vivo to evaluate the properties necessary for gut barrier protection. Cell surface hydrophobicity (CSH), autoaggregation (AA), cell adhesion, and lipopolysaccharide (LPS)-binding properties were evaluated. In addition, the suppressive effect on LPS-induced interleukin (IL)-8 expression was investigated in HT-29 cells. To identify optimal conditions for CBT LA1 to adhere to HT-29 cells, CBT LA1 cells were heat-treated at 80, 85, 90, 95, 100, or $121^{\circ}C$ for 10 min; cells treated at $80^{\circ}C$ for 10 min showed the highest adhesion. Heat-killed bacteria at $80^{\circ}C$ showed higher levels of LPS-binding, CSH, AA, adhesion to HT-29, and suppression of IL-8 expression than did live CBT LA1. In vivo imaging was performed to evaluate the ability of live or heat-killed CBT LA1 to remove LPS from the intestine in a rat model of infection. At 16 h after infection, fluorescence from FITC-conjugated LPS had mostly disappeared from the intestine of the rats administered with live or heat-killed CBT LA1; the effect was greater with heat-killed CBT LA1 at $80^{\circ}C$. These results suggest that heat-killed CBT LA1 as well as its live form can be applied as a pharmabiotic for protection of the gut barrier.

• Journal of Microbiology and Biotechnology
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• v.17 no.7
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• pp.1120-1126
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• 2007

This study was performed to screen probiotic bifidobacteria for their ability to bind and neutralize lipopolysaccharides (LPS) from Escherichia coli and to verify the relationship between LPS-binding ability, cell surface hydrophobicity (CSH), and inhibition of LPS-induced interleukin-8 (IL-8) secretion by HT-29 cells of the various bifidobacterial strains. Ninety bifidobacteria isolates from human feces were assessed for their ability to bind fluorescein isothiocyanate (FITC)-labeled LPS from E. coli. Isolates showing 30-60% binding were designated LPS-high binding (LPS-H) and those with less than 15% binding were designated LPS-low binding (LPS-L). The CSH, autoaggregation (AA), and inhibition of LPS-induced IL-8 release from HT-29 cells of the LPS-H and LPS-L groups were evaluated. Five bifidobacteria strains showed high levels of LPS binding, CSH, AA, and inhibition of IL-8 release. However, statistically significant correlations between LPS binding, CSH, AA, and reduction of IL-8 release were not found. Although we could isolate bifidobacteria with high LPS-binding ability, CSH, AA, and inhibition of IL-8 release, each characteristic should be considered as strain dependent. Bifidobacteria with high LPS binding and inhibition of IL-8 release may be good agents for preventing inflammation by neutralizing Gram-negative endotoxins and improving intestinal health.

• KSBB Journal
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• v.32 no.2
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• pp.153-159
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• 2017

The purpose of this study was to investigate the probiotic properties of lactic acid bacterial strains isolated from human feces. The properties were tested on the basis of guideline for probiotic selection protocol such as tolerance for acid or bile salt, autoaggregation, antibiotic resistance, and antimicrobial activity. Total 25 lactic acid bacteria were isolated from human feces, and their antibacterial activity was tested against Staphylococcus aureus, Escherichia coli, E. coli O157:H7, Vibrio parahaemolyticus, V. alginolyticus using an agar diffusion assay. Among them, 4 selected strains were identified by analysis of their 16S rRNA, as Lactobacillus rhamnosus MG316, L. acidophilus MG501, L. reuteri MG505, and L. gasseri MG570. Results show that resistance to low pH and bile salts. Also, the selected strains were resistant to bile acid up to 3% and their autoaggregation rates were as high as 60%. All strains tested were resistance to nalidixic acid and kanamycin.

• Journal of Life Science
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• v.33 no.4
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• pp.334-342
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• 2023

Forty species of lactic acid bacteria isolated from gajami sikhae were identified as Lactobacillus plantarum, Leuconostoc mesenteroides, Lactobacillus brevis, and Weissella paramesenteroides. 10 of the 40 strains were selected and used for the test. In this study, experiments such as those using acid and artificial gastric juice resistance, bile acid resistance, autoaggregation, coaggregation, and cell surface hydrophobicity were conducted to utilize lactic acid bacteria separated from gajami sikhae as probiotics. The separated lactic acid strains showed high survival rates through displaying resistance to acidic and artificial gastric juices; L. plantarum GS11 showed the best resistance. Also, as a result of a measurement of bile acid resistance, all lactic acid bacteria stocks showed survival of more than 100% with a probiotic number of 10⁸ to 10⁹ log CFU/ml. After evaluating cohesion to indirectly measure cell surface adhesion, autoaggregation ability was shown to be more than 46%. Measurement of xylene adhesion for cell surface hydrophobicity evaluation revealed better cell adhesion than B. subtilis, which has 32.2% hydrophobicity in isolated lactic acid strains. Antibacterial force measurement found antibacterial activity in lactic acid bacteria, excluding L. plantarum GS12 and L. plantarum GS13. Therefore, it was judged that lactic acid bacteria separated from gajami sikhae could be used as probiotics with various probiotic properties.

• Food Science and Biotechnology
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• v.17 no.2
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• pp.257-261
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• 2008

Previous studies have suggested a possible correlation between cell surface hydrophobicity (CSH) and stress tolerance in Bifidobacterium. In this study, the relationship was examined between CSH and environmental stress tolerance in Lactobacillus spp. By measuring the adhesion to hexadecane, 2 Lactobacillus fermentum strains- KLB 261 and KLB 231 were found to have high and low CSH, respectively. To measure their tolerance to various stresses, cells were subjected to salt (2 M NaCl), acid (pH 2), $H_2O_2$ (0.01 %, v/v), ethanol (20%, v/v), heat ($60^{\circ}C$), and cold ($-20^{\circ}C$). Compared with KLB 231, the hydrophobic KLB 261 was found to be much more resistant to the various stresses examined. After being subjected to different stresses for a period of time, KLB 261 and KLB 231 showed 50 and 0% survivability in 2 M NaCl, 108.2 and 0.6% in 0.01 %(v/v) $H_2O_2$, 40.2%(v/v), and 3.7% at $60^{\circ}C$ incubation, 4 and 0.6% at $-20^{\circ}C$, 12.9 and 0.1 % in pH 2, 33.8 and 0.2% in 20%(v/v) ethanol, respectively. Autoaggregation test and morphological observation were also conducted in an attempt to explain these differences. These results suggested that high CSH could strengthen the stress tolerance of lactobacilli.

• Journal of Microbiology and Biotechnology
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• v.27 no.11
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• pp.1971-1982
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• 2017

Piliated Lactobacillus rhamnosus (pLR) strains possess higher adherent capacity than non-piliated strains. The objective of this study was to isolate and characterize probiotic pLR strains in human fecal samples. To this end, mouse polyclonal antiserum (anti-SpaA) against the recombinant pilus protein (SpaA) of L. rhamnosus strain GG (LGG) was prepared and tested for its reactivity and specificity. With the anti-SpaA, a method combining the de Man, Rogosa, and Sharpe (MRS) agar plating separation and colony immunoblotting (CIB) was developed to isolate pLR from 124 human fecal samples. The genetic and phenotypic characteristics of the resultant pLR isolates were compared by randomly amplified polymorphic DNA (RAPD) fingerprinting, and examination of adhesion to Caco-2 cells, hydrophobicity, autoaggregation, and in vitro gastrointestinal tolerance. Anti-SpaA specifically reacted with three pLR strains of 25 test strains, as assessed by western blotting, immunofluorescence flow cytometry, and immunoelectron microscopy (IEM) assays. The optimized MRS agar separation plus anti-SpaA-based CIB procedure could quantitatively detect $2.5{\times}10^3CFU/ml$ of pLR colonies spiked in $10^6CFU/ml$ of background bacteria. Eight pLR strains were identified in 124 human fecal samples, and were confirmed by 16S RNA gene sequencing and IEM identification. RAPD fingerprinting of the pLR strains revealed seven different patterns, of which only two isolates from infants showed the same RAPD profiles with LGG. Strain PLR06 was obtained with high adhesion and autoaggregation activities, hydrophobicity, and gastrointestinal tolerance. Anti-SpaA-based CIB is a rapid and inexpensive method for the preliminary screening of novel adherent L. rhamnosus strains for commercial purposes.

• Environmental Engineering Research
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• v.22 no.1
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• pp.61-74
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• 2017

This study investigated the effect of various cations ($Ca^{2+}$, $Mg^{2+}$, $Al^{3+}$, $Mn^{2+}$, $Zn^{2+}$) on the autoaggregation (AAg) and surface hydrophobicity (SHb) of three different bacteria (Brevibacillus panacihumi strain (ZB1), Lysinibacillus fusiformis strain (ZB2) and Enterococcus faecalis strain (ZL)) using a 2-level factorial design. The AAg ratio was measured from the changes in the absorbance of the media. Results show that ZB2 had maximum AAg for the three bacteria investigated. A microscopic clustering of cells was observed when $Ca^{2+}$ was added to ZB2. The AAg was in the range of 62%, 58% and 34% for ZB2, ZB1 and ZL, respectively and correlated to the SHb. The aggregation and SHb of the microbial cells increased with increasing ionic strength due to the repulsive steric or overlap forces between the polymer covered surfaces. $Ca^{2+}$ demonstrated a more significant effect on aggregation and SHb of microbial cells due to an attractive binding force.

• KSBB Journal
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• v.32 no.3
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• pp.199-205
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• 2017

The aim of this study was to investigate probiotic characteristics and fermentation profile of selected lactic acid bacteria (LAB) isolated from traditional fermented foods. Antibacterial activity against various pathogens, acid and bile salt tolerance, cell hydrophobicity, and antibiotic resistance were examined. 16S rRNA sequencing was carried out to identify eight presumptive LAB isolates. In general, all identified LAB (Enterococcus faecium MG89-2, Lactobacillus plantarum MG207, L. paracasei MG310, L. casei MG311, Streptococcus thermophilus MG510, L. bulgaricus MG515, L. helveticus MG585, and L. fermentum MG590) showed strong antimicrobial activity. Also, the selected strains were resistant to bile acid up to 3% and their autoaggregation rates were as high as 60%. All selected strains tested were sensitive to chloramphenicol, tetracycline, and ampicillin, whereas resistant to nalidixic acid and kanamycin.

• KSBB Journal
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• v.31 no.1
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• pp.40-45
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• 2016

Vaginitis, also known as vaginal infection and vulvovaginitis, is an inflammation of the vagina and possibly vulva. The three main kinds of vaginitis are bacterial vaginosis, vaginal candidiasis, and trichomoniasis. The purpose of this study was to characterize Lactobacillus plantarum MG989 and L. fermentum MG901 isolated from the vaginas of healthy Korean women in terms of their inhibitory activity against the vaginitis associated pathogens such as Gardnerella vaginalis and Candida albicans. Co-culture experiments showed that the two Lactobacillus strains MG989 and MG901 significantly reduced the viability of G. vaginalis and C. albicans. Also, the two strains were resistant to bile acid up to 1% and their autoaggregation rates were as high as 83.33%. Further studies are underway to demonstrate that the two strains can be applied as pharmaceutical agents for recovering healthy vaginal ecosystem.