• Asian-Australasian Journal of Animal Sciences
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• v.15 no.12
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• pp.1798-1807
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• 2002

This study was conducted to investigate lactobacillus salivarius subsp. salivarius having probiotic properties to be used as the health adjuncts with fermented milk products. Acid- and bile-tolerant lactobacillus salivarius subsp. salivarius was isolated with lactobacilli MRS broth from faeces of 80 healthy persons (infants, children and adults). It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at 37$^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by 0.9-1.0% at 37$^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced 23-38% of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 h at 37$^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with 15-25 mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

• Journal of Dairy Science and Biotechnology
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• v.30 no.1
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• pp.45-53
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• 2012

Lactobacillus acidophilus isolated from the feces of a 7-day-old breast-fed infant was characterized to examine the scope of its commercial use. Forty-three Lactobacillus strains, which could grow at pH 5.5, were isolated. From these Lactobacillus isolates, 14 Lactobacillus strains were selected, which demonstrated more than 80% viability and homofermentative lactic fermentation. Finally, 9 L. acidophilus strains (NB 201~NB 209) were identified as candidate strains based upon biochemical properties, carbohydrate utilization, and cellular fatty acid composition. L. acidophilus isolates demonstrated a survival rate of more than 80% when exposed to pH 2.5 for 2 h. In particular, L. acidophilus NB 204 showed a strong acid tolerance, with a 71% survival rate even at pH 2.0. L. acidophilus isolates also manifested high bile acid tolerance; more than 87% of the cells survived on agar containing 1% bile extract, except for L. acidophilus NB 206, which showed a 73% survival rate. All L. acidophilus isolates were confirmed to have proteolytic activity; L. acidophilus NB 204 and NB 209 yielded higher levels of TCA-soluble peptides and free amino acids. The ${\beta}$-galactosidase activity of the L. acidophilus isolates was in the range of 1.97~2.45 units/mL.

• Journal of Life Science
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• v.22 no.5
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• pp.575-581
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• 2012

This study was conducted to develop an economical optimum medium composition for the mass production of $Lactobacillus$ $plantarum$ and $Lactobacillus$ $reuteri$, livestock probiotics. Medium ingredient factors were selected on the basis of MRS broth composition, and the 15 ingredient variables were as follows: sucrose, glucose, molasses, yeast extract, corn steep liquor, soy peptone, dipotassium phosphate, manganese chloride, magnesium chloride, tween 80, sodium chloride, sodium acetate, ammonium citrate, sodium sulphate, and ferrous sulphate. The Plackett Burman design, consisting of 20 runs, was employed for the analysis of ingredient effects on cell growth of $L.$ $plantarum$ and $L.$ $reuteri$. As a result, sucrose, glucose, molasses, yeast extract, corn steep liquor, soy peptone, sodium acetate, and ammonium citrate positively influenced the growth of $L.$ $plantarum$. Additionally, yeast extract, soy peptone, $K_2PHO_4$, and tween 80 positively influenced the growth of $L.$ $reuteri$. Positive effects were found from sucrose, yeast extract, and soy peptone in the integrated analysis of the effects of both $L.$ $plantarum$ and $L.$ $reuteri$. Finally, effective medium components for both strains were found as follows: sucrose (20.0 g/l), glucose (5.0 g/l), soy peptone (11.0 g/l), yeast extract (5.0 g/l), $K_2PHO_4$ (0.2 g/l), $CH_3COONa$ (2 g/l), and $MgCl_2$ (0.02 g/l).

• Journal of The Korean Society of Grassland and Forage Science
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• v.35 no.4
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• pp.321-326
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• 2015

This study was performed to analyze the resident microbial flora and the effects of probiotic inoculation on the fermentation characteristics of whole grain naked barley (Hordeum Vulgare L.) with the goal of evaluating the possibility of utilization as fermented feedstuff. Naked barley grains were harvested 35 days after heading, and the microbial flora was analyzed using MALDI-TOF mass spectrometer. After inoculation of commercial microbes to the naked barley grain (BT), the pH and number of bacteria, such as aerobic bacteria, lactic acid bacteria, yeast and E. coli, were measured and compared with the non-inoculated control (BC). A total of 122 colonies was isolated from the naked barley grain and the most popular bacteria species was Staphylococcus xylosus (n = 30, 24.59%). The pH value decreased more rapidly in BT than in BC, and was significantly lower after 7 days of fermentation at $4.33{\pm}0.02$ and $4.83{\pm}0.01$, respectively. The number of aerobic bacteria, lactic acid bacteria and yeast showed an increasing trend within the first 7 days of fermentation, however, their numbers decreased at 28 and 42 days of fermentation. The population of lactic acid bacteria in BT was higher than in BC, but there was no significant different at 7 days of fermentation, with respective levels of $9.24{\pm}0.20$ and $9.01{\pm}0.10logCFU/g$ (p>0.05). The initial number of E. coli was very high in the naked barley grain but subsequently decreased significantly. After 7 days of fermentation, E. coli was not detected in either BT or BC samples. From these results, it appears that the fermentation of naked barley grain proceeded adequately after 7 days, and that fermentation contributes to the safety of naked barley grain during storage.

• Microbiology and Biotechnology Letters
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• v.40 no.4
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• pp.389-395
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• 2012

T-RFLP (terminal restriction fragment length polymorphism) analysis, one of the most highly adopted culture-independent microbial community analysis methods, was applied to evaluate the colonization of probiotics in experimental animal gut. Lactic acid bacteria that exhibited cinnamoyl esterase activity were isolated from Korean fermented vegetables and identified by 16S ribosomal RNA sequence analysis. Lactobacillus plantarum KK3, which demonstrated high chlorogenic acid hydrolysis by cinnamoyl esterase activity, and acid/bile salt resistances, was cultured, freeze-dried, and fed to mice and the microbiota in their feces were monitored by T-RFLP analysis. The T-RF of L. plantarum was detected in the feces of mice after the start of administration and lasted at least 31 days after the initial 7 day feeding. T-RFLP analysis was considered a useful tool to evaluate the gut colonization of probiotic L. plantarum. In order to prove that L. plantarum was from viable cells, we reisolated L. plantarum in the feces using cinnamoyl esterase activity media as the screening step. The colonization of L. plantarum KK3 in the mouse gut was confirmed by this research.

• Journal of Animal Science and Technology
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• v.50 no.5
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• pp.667-676
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• 2008

This experiment was conducted to investigate the effects of dietary supplementation of various Lactobacilli strains on growth performance, intestinal microflora, ileal villi development and ileal mucosal surface of broiler chickens. Six hundred 1-d-old male chicks(Avian) were randomly divided into five groups of four replicates, each replicates containing 30 birds and fed corn-soybean meal diets containing Latobaillus were isolated from cecum of chichens at 107cfu/g diet for five weeks. The treatments were control(antibiotics-free diet), Lactobacillus crispatus avibro1(LCB), Lactobacillus reuteri avibro2(LRB), Lactobacillus crispatus avihen1(LCH) or Lactobacillus vaginalis avihen2(LVH). The body weight gain in groups fed Lactobacillus significantly improved as compared to those of control(P<0.05). Feed intake was not statistically different among the groups. The number of Lactobacillus in ileum and cecum of chicks fed various Lactobacillus tended to be increased or significantly increased as compared to those of the control(P<0.05), but there was difference by age of chicks and species of Lactobacillus. The number of yeast was significantly increased in cecum and ileum at three weeks old chicks fed Lactobacilli compared with the control(P<0.05). The anaerobes’ number of ileum and cecum tended to increase or significantly increased in Lactobacillus treatments compared with the control(P<0.05). The ileal villi length extended greatly at three weeks of age in groups fed Lactobacillus compared with the control(P<0.05). The length of ileal villi in chicks fed Lactobacillus was continuously increased up to five weeks of age, but did not increased in the control(P<0.05). Lactobacillus was found on ileal mucosal surface. And ileal mucosal surface was maintained better with Latobacillus feeding. These results suggest the possibility that cecal Lactobacillus of chickens could be used as probiotics by improving the growth performance and promoting development of ileal villi in broiler chicks.

• Journal of Life Science
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• v.27 no.2
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• pp.172-179
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• 2017

The fermentation of Panax ginseng can yield many compounds from ginsenosides that have a wide variety of biological functions. Lactic acid bacteria (LAB) strains are capable of converting ginsenosides. The purposes of this study were to: (1) characterize Enterococcus faecium SA5, an isolated LAB from Mongolian mare milk, (2) identify the existence of extracellular ${\beta}$-glucosidase activity in the milk, and (3) ascertain if the ${\beta}$-glucosidase has the capacity of converting ginsenoside in Korean ginseng. The results revealed that E. faecium SA5 was acid-resistant, bile salt-resistant, and has antibiotic activities against 4 pathogenic microorganisms (Salmonella typhimurium KCTC 3216, Listeria monocytogenes KCTC 3710, Bacillus cereus KCTC 1012, Staphylococcus aureus KCTC 1621). In addition, E. faecium SA5 had tolerance against some antibiotics such as colistin, gentamycin and neomycin. It was also found that E. faecium SA5 possessed bile salt hydrolase activity, which could lower blood cholesterol level. When incubated in 10% (w/v) skim milk as a yogurt starter, E. faecium SA5 caused to decrease pH of the medium as well as increase in viable cell counts. Using TLC and HPLC analysis on the samples incubated in MRS broth, our study confirmed that E. faecium SA5 can produce ${\beta}$-glucosidase, which was capable of converting ginsenoside $Rb_1$ into new ginsenosides $Rg_3-s$ and $Rg_3-r$. It was concluded that E. faecium SA5 possessed a potential of probiotic activity, which could be applied to yogurt manufacture as well as ginsenoside conversion in ginseng.

• Journal of the Korean Applied Science and Technology
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• v.34 no.4
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• pp.807-817
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• 2017

The present study was undertaken to investigate the effect of fermented earthworm cast feed prepared by using three probiotic mixture with Bacillus subtilis strain on meat quality and growth performance of broiler chickens. A total of 240 one-day-old broiler chickens were randomly assigned to four treatments: 1) control, 2) 0.2% commercial product contain strain of Lactobacillus fermentum (CP), 3) 0.3% fermented earthworm cast feed (FECF3); and 4) 0.5% fermented earthworm cast feed (FECF5). Mean body weight gain, thymus, spleen, and F-bursal weight of birds fed with FECF were higher than those of birds fed with control or CP. Serum IgG levels were higher in birds fed with FECF than those in birds fed with CP or control. Cecal Lactobacillus counts were higher whereas Escherichia, Salmonella, Coliform bacteria, and total aerobic bacteria counts were lower in FECF groups than those in CP or control group. Water holding capacities in FECF groups were higher than those in CP or control group. Ratios of n-6/n-3 in chicken meat were lower in FECF groups compared to those in CP or control group.

• Journal of Animal Science and Technology
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• v.64 no.1
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• pp.166-182
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• 2022

Deer antler velvet is widely used in traditional medicine for its anti-aging, antioxidant, and immunity-enhancing effects. However, few studies have reported on the discovery of probiotic strains for deer antler fermentation to increase functional ingredient absorption. This study evaluated the ability of probiotic lactic acid bacteria to enhance the concentrations of bioactive molecules (e.g., sialic acid and gamma-aminobutyric acid [GABA]) in extracts of deer antler velvet. Seventeen strains of Lactobacillus spp. that were isolated from kimchi and infant feces, including L. sakei, L. rhamnosus, L. brevis, and L. plantarum, and those that improved the life span of Caenorhabditis elegans were selected for evaluation. Of the 17 strains, 2 (L. rhamnosus LFR20-004 and L. sakei LFR20-007) were selected based on data showing that these strains increased both the sialic acid and GABA contents of deer antler extract after fermentation for 2 d and significantly improved the life span of C. elegans. Co-fermentation with both strains further increased the concentrations of sialic acid, GABA, and metabolites such as short-chain fatty acids and amino acids. We evaluated the biological effects of the fermented antler velvet (FAV) on the antibacterial immune response in C. elegans by assessing worm survival after pathogen infection. The survival of the C. elegans conditioned with FAV for 24h was significantly higher compared with that of the control worm group fed only normal feed (non-pathogenic E. coli OP50) exposed to E. coli O157:H7, Salmonella typhi, and Listeria monocytogenes. To evaluate the protective effects of FAV on immune response, cyclophosphamide (Cy), an immune-suppressing agent was treated to in vitro and in vivo. We found that FAV significantly restored viability of mice splenocytes and immune promoting-related cytokines (interleukin [IL]-6, IL-10, inducible nitric oxide synthase [iNOS], interferon [IFN]-γ, and tumor necrosis factor [TNF]-α) were activated compared to non-fermented deer antlers. This finding indicated the protective effect of FAV against Cy-induced cell death and immunosuppressed mice. Taken together, our study suggests that immune-promoting antler velvet can be produced through fermentation using L. rhamnosus LFR20-004 and L. sakei LFR20-007.

• Biomolecules & Therapeutics
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• v.17 no.3
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• pp.276-281
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• 2009

Lactic acid bacteria (LAB) are considered as probiotics with immunostimulatory property. In this study, we investigated the molecular mechanism of its immunostimulating potency on macrophages using combined preparation of LAB (cpLAB). cpLAB is able to strongly stimulate nitric oxide (NO) production as well as inducible NO synthase (iNOS) expression from macrophage-like RAW264.7 cells. The cpLAB-induced NO release seemed to be mediated by extracellular signal-regulated kinase (ERK) but not p38 and C-Jun N-terminal kinase (JNK), since U0126, an ERK inhibitor, clearly suppressed NO production. cpLAB significantly diminished the binding of toll like receptor (TLR)-2 antibody up to 25%, implying that cpLAB-mediated activation of macrophages may be required for the functional activation of TLR-2, but not TLR-4. Therefore, our data suggest that cpLAB may directly allow macrophages to immunostimulating potency via activation of TLR-2 and ERK.