• Asian-Australasian Journal of Animal Sciences
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• 제14권8호
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• pp.1170-1178
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• 2001

This study was conducted to isolate lactobacilli having characteristics to be used as health adjuncts with fermented milk products. Acid tolerant strains were selected in Lactobacilli MRS broth adjusted to pH 4.0 from human faeces. Bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. Microhemagglutination tests using swine erythrocytes were performed to select lactobacilli having adherence properties to survive in the intestinal tract. By examination of these characteristics the strain Nam 27, which was isolated from adult faeces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing.

• 대한수의학회지
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• 제40권4호
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• pp.701-706
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• 2000

This study was carried out to select the lactic acid bacteria(Lactobacillus, Streptococcus and Bifidobacterium) and yeast for probiotic use in pigs. Acid-tolerant 536 strains were isolated from the feces of 30 pigs. To select useful strains, the first screened strains were treated with strong acid solution(pH 2.5 to 3.0) for 3 hours and subsequentely treated with the anaerobic diluent solution containing 0.15% Oxgall for 3 hours. Among these strains, 151 strains showed strong tolerance to both acid and bile. Lactobacillus and Streptococcus tolerant to the acid and bile were treated with heat at $80^{\circ}C$ for 15 min, and at $70^{\circ}C$ for 5 min in Bifidobacterium and yeast. As a result of heat treatment, 38 strains were obtained as heat-tolerant strains. All of heat-tolerant strains were tested for antibiotic resistance against virginiamycin, sulfathiazole, aureomycin, neomycin, linsmycin, tiamulin and ASP250 which were used as feed additives for growth promotion in pigs. Finally, one strain each from Lactobacillus, Streptococcus, Bifidobacterium and yeast that showed resistance to acid, bile, heat and antibiotics was selected for probiotic use in pigs.

• Food Science and Biotechnology
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• 제27권6호
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• pp.1747-1754
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• 2018

The objective of this study was to evaluate probiotic effects of two Lactobacillus plantarum strains (GBL16 and 17) isolated from black raspberry. Results revealed that the number of GBL16 was gradually decreased as bile salt concentration was increased from 0.3 to 1%. However, GBL17 did not show any difference when GBL17 was applied to 1% bile salt, and it indicates that GBL17 is more tolerant to bile salt than GBL16. GBL17 exhibited higher heat resistance and adhesion ability to Caco-2 cells than GBL16. Regarding gut microbiome, no significant change in the number of total bacteria in intestines of mice after treatment with GBLs was determined. However, the combination of GBL16 and GBL17 significantly increased the number of total bacteria in intestines of mice after they were orally administered. Therefore, the results suggest that both GBL16 and 17 strains could be one of major probiotics that can improve human gut health.

• Journal of Microbiology and Biotechnology
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• 제30권5호
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• pp.739-748
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• 2020

In this study, a method of heat adaptation was implemented in an attempt to increase the upper thermal threshold of two Streptococcus thermophilus found in South Korea and identified the alterations in membrane fatty acid composition to adaptive response to heat. In order to develop heat tolerant lactic acid bacteria, heat treatment was continuously applied to bacteria by increasing temperature from 60℃ until the point that no surviving cell was detected. Our results indicated significant increase in heat tolerance of heat-adapted strains compared to the wild type (WT) strains. In particular, the survival ratio of basically low heat-tolerant strain increased even more. In addition, the strains with improved heat tolerance acquired cross protection, which improved their survival ratio in acid, bile salts and osmotic conditions. A relation between heat tolerance and membrane fatty acid composition was identified. As a result of heat adaptation, the ratio of unsaturated to saturated fatty acids (UFA/SFA) and C18:1 relative concentration were decreased. C6:0 in only heat-adapted strains and C22:0 in only the naturally high heat tolerant strain were detected. These results support the hypothesis, that the consequent increase of SFA ratio is a cellular response to environmental stresses such as high temperatures, and it is able to protect the cells from acid, bile salts and osmotic conditions via cross protection. This study demonstrated that the increase in heat tolerance can be utilized as a mean to improve bacterial tolerance against various environmental stresses.

• 미생물학회지
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• 제47권3호
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• pp.231-240
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• 2011

숙성된 갓김치에서 분리된 유산균을 pH 2.5에서 2시간 반응시킨 후 0.3% oxgall 존재 하에서 3시간 배양시킨 결과 MLK11, MLK22, MLK27, MLK41 및 MLK67 균주들은 $10^5$ CFU/ml 이상의 균수를 유지하여 높은 저항성을 보인 반면, MLK53 균주는 대부분의 균수가 사멸되어 매우 민감한 것으로 나타났다. 담즙산에 대한 내성이 강한 균주들 대부분은 복합 담즙산의 탈포합능이 있었으며, MLK22와 MLK67 균주는 sodium glycocholate로부터 3.5 mM 이상의 cholic acid을 유리시켜 가장 높은 탈포합능을 보인 반면, MLK13과 MLK41은 각각 1.35와 1.16 mM 정도 낮은 양의 cholic acid를 유리시켰다. 특히, MLK22와 MLK67의 탈포합능은 sodium taurocholate 혹은 포합담즙산 혼합물 보다는 sodium glycoholate 존재 하에서 더 높게 나타났다. 게다가 sodium glycocholate와 sodium taurocholate으로부터 MLK22와 MLK67이 생산하는 bile salt hydrolase (BSH)의 활성은 정지기 초기에 최대를 이르렀고 MLK67 보다는 MLK22의 BSH 활성이 다소 높았다. 한편, 실험 균주들의 콜레스테롤 제거능은 5.22-39.16 ${\mu}g$/ml로 균주별 유의적인 차이가 있었으며(p<0.05), 그 중에서 MLK67 균주는 0.3% oxgall, cholic acid 및 taurocholic acid로부터 가장 높은 콜레스테롤 동화능을 나타내었다. 따라서 실험 균주 중 높은 내산성과 내담즙성을 가지며, 담즙산 탈포합능 및 콜레스테롤 동화능이 유의하게 높은 MLK67 균주는 probiotic 균주로서의 가능성이 있는 것으로 간주되어 이를 생화학적 특성과 당분해능 및 염기서열 분석에 의해 동정한 결과 Pediococcus pentosaceus MLK67로 확인되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제22권1호
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• pp.119-123
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• 2009

In vitro acid- and bile-tolerant lactic acid bacteria isolated and identified from Indonesian traditional fermented milk dadih might be considered as potential probiotic strains after further characterization with animal models, especially for their therapeutic properties. Five dadih lactic bacteria isolates each had moderate survival rate for 2 h at pH 2.0, as well as bile tolerance. The aim of this research was to identify candidate probiotic lactic bacteria among indigenous dadih lactic isolates originated from Bukit Tinggi, West Sumatra, especially their in vivo antimutagenic property. Milk cultured with Enterococcus faecium IS-27526 significantly lowered fecal mutagenicity of rats as compared to the control group, skim milk, and milk cultured with L. plantarum IS-20506. These results suggest that Enterococcus faecium IS-27526 may serve as a potential probiotic strain with its antimutagenicity.

• 대한의생명과학회지
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• 제11권4호
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• pp.539-543
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• 2005

Elevated level of serum cholesterol in humans is a risk factor correlated with the development of coronary heart disease. We have screened lactic acid bacteria from pig and human feces for the development of probiotics which have an anti-cholesterol effect. We have used special media to isolate only lactic acid bacteria and they were subjected to the experiments such as oxgal test, carbohydrate fermentation test. Results from the acid tolerance test and growth test in the presence of oxgal demonstrated that some strains would likely survive in thuman stomach, where acidity is high, and in small intestine, where bile fluid is present. In conclusion, we were able to screen lactic acid bacteria which were tolerant against bile acid and supposed to be prominent to lower the cholesterol level in human serum.

• Asian-Australasian Journal of Animal Sciences
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• 제16권11호
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• pp.1680-1685
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• 2003

Antimutagenicity of milk cultured with lactic acid bacteria isolated from dadih on the mutagenicity of heated salty and sweet tauco was examined using streptomycin dependent (SD) 510 strain of Salmonella typhimurium TA 98 as a tester culture. Cultured milk samples exhibited widely antimutagenic activity against mutagenic heated salty and sweet tauco. Lc. lactis subsp. lactis R-22, Lc. lactis subsp. casei R-35, Lc. lactis subsp. casei R-52 and E. faecalis subsp. liquefaciens R-55 exhibited no inhibitory effect on the mutagenic heated salty tauco. Mutagenicity of heated sweet tauco was inhibited by cultured milks stronger than that of heated salty tauco. Milk cultured with Lc. lactis subsp. cremoris R-48, Leuc. mesentroides R-51 and Lc. lactis subsp. casei R-68 showed high inhibition against the mutagenicity of both heated salty and sweet taucos. Antimutagenic activity of the cultured milks against mutagenic heated tauco was attributed to the bacterial cells. Among the three strains which showed high antimutagenicity, only Leuc. mesentroides R-51 was tolerant to both acid and bile; so this strain can be used as probiotic in preventing the occurrence of mutagenesis caused by mutagenic heated food like tauco.

• Journal of Microbiology and Biotechnology
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• 제12권1호
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• pp.162-165
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• 2002

With the objective of identifying the commercial potential of new direct-fed microbials, several temperature-tolerant strains were isolated from cane molasses at $39^{\circ}C$ and tested for their tolerance to pH, bile salts, and a mixture of volatile fatty acids (acetic acid:propionic acid:butyric acid=6.5:2.0:1.5). It was found that the isolated strain DY 252 grew very well up to pH 2.0 and was resistant to relatively high concentrations of bile salts. Among the strains tested, DY 252 was least inhibited by the addition of volatile fatty acids to the growth medium at $39^{\circ}C$. Accordingly, it would appear that strain DY 252, identified as yeast Issatchenkia orientalis, may be a potential candidate for use as a microbial feed additive.

• Asian-Australasian Journal of Animal Sciences
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• 제16권5호
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• pp.726-731
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• 2003

The aim of this research was to identify candidate probiotic lactic bacteria among indigenous dadih lactic isolates. Dadih is an Indonesian traditional fermented milk of West Sumatra which is fermented naturally. Viability of the strain is critical in determining the capacity of lactic bacteria to induce immune stimulation as well as to colonize in the intestinal tract. Therefore, LAB are proposed to exert health promoting or probiotic effects in human, such as inhibition of pathogenic microflora, antimutagenic, and the reduction of cholesterol levels. This manuscript reports in vitro probiotic properties of indigenous dadih lactic bacteria, especially some important colonization factors in GI tract, such as lysozyme, acid and bile tolerance. Bile Salt Hydrolase (BSH) activity, spectrum of bacteriocin, and antimutagenic activity of bacterial cells were also assessed. Twenty dadih lactic isolates were screened further for their tolerance to low pH, at pH 2 and 3 as well as their bile tolerance. There were ten isolates classified as acid and bile acid tolerant, and further screened for lysozyme tolerance, BSH activity. The spectrum of bacteriocin activity of isolates was assayed using cell-free neutralized supernatants by agar spot test against variety of pathogens. Lc. lactis subsp. lactis IS-10285, IS-7386, IS-16183, IS-11857 and IS-29862, L. brevis IS-27560, IS-26958 and IS-23427, Leu.mesen.mesenteroides IS-27526, and L. casei IS-7257 each has good survival rate at low pH values and in the presence of lysozyme, and short lag time in the presence of 0.3 % oxgall. Lc. lactis subsp. lactis IS-11857 and IS-29862 each has high BHS activity, Lc. lactis subsp. lactis IS-10285 and IS-16183 each had a positive spectrum of bacteriocin activity against E. coli 3301 and Lysteria monocytogenes ATCC 19112, while L. brevis IS-26958 has high BHS activity as well as positive spectrum of bacteriocin against E. coli 3301, Lysteria monocytogenes ATCC 19112, and S. aureus IFO 3060. All of the ten dadih lactic strains performed in vitro acid and bile tolerance, indicating a possibility to reach the intestine alive, and display probiotic activities.