• 한국축산식품학회:학술대회논문집
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• 한국축산식품학회 2004년도 정기총회 및 제33차 춘계 학술대회
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• pp.89-119
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• 2004

This study was conducted to isolate lactobacilli having probiotic 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 80 healthy persons (infants, children and adults). And bile tolerant strains were examined in Lactobacilli MRS broth in which 1.0% bile salt was added. By estimation above characteristics, the strains No. 27, which was isolated from adult feces, was selected and identified as Lactobacillus salivarius subsp. salivarius based on carbohydrate fermentation and 16S rDNA sequencing. 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 $0.9{\sim}1.0%$ 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 $23{\sim}38%$ of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 hours at $37^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with $15{\sim}25$ 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. Purified ${\alpha}$-galactosidase was obtained by DEAE-Sephadex A-50 ion exchange chromatography, Mono-Q ion exchange chromatography and HPLC column chromatography from L. salivarius subsp. salivarius 27. The specific activity of the purified enzyme was 8,994 units/mg protein, representing an 17.09 folds purification of the original cell crude extract. The molecular weight of enzyme was identified about 53,000 dalton by 12% SDS-PAGE. Optimal temperature and pH for activity of this enzyme were $40^{\circ}C$ and 7.0 respectively. The enzyme was found to be stable between 25 and $50^{\circ}C$. ${\alpha}$-galactosidase activity was lost rapidly below pH 5.0 and above pH 9.0. This enzyme was liberated galactose from melibiose, raffinose, and stachyose, and also the hydrolysis rate of substrate was compound by HPLC. 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.

• 한국축산식품학회지
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• 제26권1호
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• pp.127-135
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• 2006

우유와 두유를 2:1로 혼합하고 홍삼 추출물을 첨가하여 발효유를 제조하였고 2종류의 스타터를 사용하여 발효 특성을 시험하였다. 2종류의 스타터를 사용한 모든 처리구에서 배양 15시간 후 pH는 $3.90{\sim}3.94$ 사이로 나타나 산 생성 촉진 효과가 있었고, 산 생성은 홍삼 추출물 첨가량이 증가함에 따라 조금씩 증가되는 것으로 나타났다. 홍삼 추출물 첨가량이 증가할수록 생균수는 증가하여 Lactobacillus acidophilus KCTC 3150를 스타터로 사용한 균에서 1.0% 홍삼 추출물 첨가량은 $6.26{\times}10^8\;cfu/mL$로서 최대 균수를 나타내었으며, 배양 12시간 이후에 대조구에서는 감소하는 반면 홍삼 추출물 첨가구에서는 젖산균수는 계속해서 최대균수를 유지하고 있었다. 유기산 생성은 lactic acid인 경우 Lactobacillus acidophilus KCTC3150에서 1.0% 홍삼 추출물 첨가구에서 332.22 mM로 가장 높았으며 홍삼 추출물 첨가량이 증가됨에 따라 lactic acid의 생성량도 증가되었다. 당 분해율은 두 균주 모두 홍삼 추출물 첨가량이 증가함에 따라 stachylose의 분해율이 증가하였고, Lactobacillus acidophilus KCTC3150 균주의 경우 stachylose의 분해율이 최대 19%인 반면, Lactobacillus salivarius subsp. salivarius CNU27의 경우 stachylose의 최대 분해율은 54%로서 분해율이 월등히 높았다. 점도는 홍삼추출물 첨가량이 1.0% 첨가구에서 Lactobacillus acidophilus KCTC3150, Lactobacillus salivarius subsp. salivarius CNU27 각각 780 cP로 가장 높게 나타났다. 관능검사 결과는 0.2% 홍삼 추출물을 첨가하고 Lactobacillus salivarius subsp. salivarius CNU27를 스타터로 사용한 발효유가 $4.14{\pm}0.64$로 기호도가 가장 좋게 나타났다.

• 한국축산식품학회지
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• 제25권4호
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• pp.483-493
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• 2005

우유와 두유를 혼합하여 요구르트를 제조하고자 스타터에 따른 발효물의 성상을 시험하였다. 4종류의 서로 다른 스타터를 접종한 모든 처리구에서 배양 15시간 후 pH는 lactic culture 2(B. longum, L. acidophilus, S. thermophilus)를 제외한 나머지 처리구에서 $4.16\~3.75$ 사이로 나타나 발효 촉진효과가 있었고, 산 생성은 우유와 두유의 홉합비가 2:1이고 lactic culture 1 [Lactobacillus delbrueckii ssp. bulgaricus(LB12), Streptococcus salivarius ssp. thermophilus(ST36)]을 스타터로 사용한 요구르트에서 가장 높게 나타났다. 생균수는 1:3 비율에서 15시간째 Lactobacillus salivarius subsp. salivarius CNU27을 스타터로 사용한 균에서 $2.17\times10^9 cfu/mL$로서 최대 균수를 나타내었다. 두유 혼합요구르트에서 생산된 유기산의 함량을 측정한 바, 유기산 생산 중 lactic acid인 경우 lactic culture 1 [Lacto-bacillus delbrueckii ssp. bulgaricus(LB12), Streptococcus salivarius ssp. thermophihus(ST36)]을 스타터로 사용했을 때 1:1에서 412.52 mM로 가장 높았다. Acetic acid인 경우는 혼합비 3:1에서 lactic culture 2(B. longum, L. acidophilus, S. thermophilus)가 394.01mM로 가장 높게 생산되었다. 당 분해율은 혼합비 1:3에서 Lactobacillus salivarius subsp. salivarius CNU27를 스타터로 사용한 요구르트에서 대조구와 비교해 stachyose가 $53.92\%$로 가장 많이 분해되었다. 점도는 두유 혼합비가 가장 높은 1:3 처리구에서 $1,300\~1,660 cP$ 로 가장 높게 나타났다. 관능검사 결과는 2:1 혼합비에서 Lactobacillus acidophilus KCTC3150를 스타터로 사용한 요구르트가 $4.17\pm0.69,$ 3: 1 혼합비에서 Lactobacillus salivarius subsp. salivarius CNU27를 스타터로 사용한 요구르트가 $4.00\pm0..58$으로 기호도가 가장 좋은 것으로 나타났다.

• 한국축산식품학회지
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• 제25권2호
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• pp.257-264
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• 2005

홍삼 추출물이 유산간균 3종(L. acidophilus, L. casei 및 L. salivarius ssp. salivarius)과 병원성 균으로서 E. coli 및 L. monocytogenes에 대한 성장 촉진과 억제 효과를 홍삼 추출물 0, 1, 2, 5와 $10\%$ 농도로 조사하였으며, 홍삼 추출물에 의한 pH 저하를 buffer제를 사용하여 조정하고 미생물의 성장을 대조구와 비교하여 조사하였다. 유산간균 3종의 경우 홍삼 추출물 $1\%$ 농도까지는 무처리구와 유사한 성장을 보였으나, $2\%$이상의 홍삼 추출물 첨가 시 농도가 높아질수록 유산균의 성장도 둔화되었다. E. coli의 경우 홍삼 추출물이 $1\%$부터 높아질수록 성장이 억제되었고, L. monocytogenes의 경우 $5\%$ 이상의 처리구에서 억제 효과가 있었다. Buffer제를 사용하여 pH를 조정하고 홍삼 추출물을 $10\%$ 첨가한 구와 pH를 조정하지 않은 구를 비교하여 보면 유산간균과 병원성 균의 성장이 두 구간에 모두 억제되므로 미생물의 억제 효과가 홍삼 추출물 중의 유기산에 의한 pH 저하가 원인이 아님을 알 수 있었으며, 이와 같은 미생물의 성장억제 효과는 홍삼 추출물 중 다른 유효성분에 의한 것으로 생각되어진다.

• Journal of Microbiology and Biotechnology
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• 제20권1호
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• pp.110-116
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• 2010

Lactic acid (LA) fermentation by Lactobacillus salivarius ATCC 11741 immobilized on loofa sponge (LS) was evaluated. To increase the surface area of LS for cell immobilization, $H_2O_2$ and chitosan were introduced as surface modifying reagents. Four chitosans of different molecular weights were separately coated on LS. All experiments were conducted in shaking flask mode at 100 rpm rotating speed and $37^{\circ}C$ with 5% $CaCO_3$ as a pH regulating agent. The effects of initial glucose concentration were investigated in the range of 20-100 g/l on LA fermentation by free cells. The results indicate that the maximum concentration of LA was produced with 50 g/l glucose concentration. The immobilized cell system produced 1.5 times higher concentration than free cells for 24 h of fermentation. Moreover, immobilized cells can shorten the fermentation time by 2-fold compared with free cells at the same level of LA concentration. At 1% (w/v) chitosan in 2% (v/v) acetic acid, the Yp/s and productivities of various molecular weights of chitosans were insignificantly different. Repeated batch fermentations showed 5 effective recycles with Yp/s and productivity in the range of 0.55-0.85 and 0.90-1.20 g/l.h, respectively. It is evident that immobilization of L. salivarius onto LS permits reuse of the system under these fermentation conditions. Scanning electron micrographs indicated that there were more intact cells on the chitosan-treated LS than on the untreated LS, thus confirming the effectiveness of the LS-chitosan combination when being utilized as a promising immobilization carrier for LA fermentation.

• Animal Bioscience
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• 제36권8호
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• pp.1209-1220
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• 2023

Objective: The use of probiotics as an alternative to antibiotics in animal feed has received considerable attention in recent decades. Lactic acid bacteria (LAB) have remarkable functional properties promoting host health and are major microorganisms for probiotic purposes. The aim of this study was to characterize LAB strains of the chicken digestive tract and to determine their functional properties for further use as potential probiotics in poultry. Methods: A total of 2,000 colonies were isolated from the ileum and cecal contents of the chickens based on their phenotypic profiles and followed by a preliminary detection for acid and bile tolerance. The selected 200 LAB isolates with exhibited well-tolerance in acid and bile conditions were then identified by sequencing the 16S rDNA gene, followed by acid and bile tolerance, antimicrobial activity, adhesion to epithelial cells and additional characteristics on the removal of cholesterol. Then, the two probiotic strains (L. ingluviei and L. salivarious) which showed the greatest advantage in vitro testing were selected to assess their efficacy in broiler chickens. Results: It was found that 200 LAB isolates that complied with all measurement criteria belonged to five strains, including L. acidophilus (63 colonies), L. ingluviei (2 colonies), L. reuteri (58 colonies), L. salivarius (72 colonies), and L. saerimneri (5 colonies). We found that the L. ingluviei and L. salivarius can increase the population of LAB and Bifidobacterium spp. while reducing Enterobacteria spp. and Escherichia coli in the cecal content of chickens. Additionally, increased concentrations of valeric acid and short chain fatty acids were also observed. Conclusion: This study indicates that all five Lactobacillus strains isolated from gut contents of chickens are safe and possess probiotic properties, especially L. ingluviei and L. salivarius. Future studies should evaluate the potential for growth improvement in broilers.

• Journal of Microbiology and Biotechnology
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• 제28권5호
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• pp.718-731
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• 2018

The beneficial effects of lactic acid bacteria (LAB) have been intensively investigated in recent decades with special focus on modulation of the host intestinal microbiota. Numerous discoveries of effective probiotics are driven by a significantly increasing demand for dietary supplements. Consequently, technological advances in the large-scale production and lyophilization are needed by probiotic-related industries for producing probiotic LAB for commercial use. Our study had a dual objective, to determine the optimum growth medium composition and to investigate appropriate cryoprotective additives (CPAs) for Lactobacillus salivarius, and compare its responses with other Lactobacillus species. The one-factor-at-a-time method and central composite design were applied to determine the optimal medium composition for L. salivarius cultivation. The following composition of the medium was established (per liter): 21.64 g maltose, 85 g yeast extract, 1.21 ml Tween 80, 6 g sodium acetate, $0.2g\;MgSO_4{\cdot}7H_2O$, $0.02g\;MnSO_4{\cdot}H_2O$, $1g\;K_2HPO_4$, $1.5g\;KH_2PO_4$, $0.01g\;FeSO_4{\cdot}7H_2O$, and 1 g sodium citrate. A cryoprotective additive combination comprising 10% (w/v) skim milk and 10% (w/v) sucrose supplemented with 2.5% (w/v) sodium glutamate was selected for L. salivarius, and its effectiveness was confirmed using culture-independent methods in the freeze-dried cells of the Lactobacillus strains. In conclusion, the optimized medium enhanced the species-specific cultivation of L. salivarius. On the other hand, the cryoprotective effects of the selected CPA mixture may also be dependent on the bacterial strain. This study highlights the necessity for precise and advanced processing techniques for large-scale production of probiotics in the food and feed industries.

• 한국식품영양학회지
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• 제1권1호
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• pp.33-40
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• 1988

A Study was conducted to investigate the effect of feeding Lactobacillus casei YS on the growing performance and gastrointestinal flora of the suckling piglets, which were delivered from 2 heads of three-way crossbred(Landrace$\times$Large White$\times$Duroc) pigs, for 4 weeks. The results from the present study was summarized as follows. Average body weight gains of feeding group was slightly better than that of control group and diarrhea was prevented by successive 7 days feeding. Population levels of lactic acid bacteria were maintained about 107 colony forming unit(cfu) per gram of the contents in both feeding and control group at upper parts of small intestine. In this part, coliform count was greatly reduced in (ceding group but not in control group. pH values of the intestinal contents were gradually decreased especially at the upper part of alimentary track of feeding group. Among lactic acid bacteria, L. salivarius, L. cases and L. fermentum were found most predominant strains in feeding group, Wheareas L. salivarius, L. acidophilus and L. cunts in control group. In the other hand, Escherichia coli recovered from scouring pigs were resistant to the drug such as streptomycin, ampicillin and sensitive to gentamycin and neomycin in vitro test.

• 대한수의학회지
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• 제51권4호
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• pp.281-288
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• 2011

Lactobacillus salivarius JWS 58 (JWS 58) and Lactobacillus plantarum JWS 1354 (JWS 1354) are isolated from duck intestine and have ability to produce bacteriocin. The objective of this study was to evaluate the immunomodulatory effects of JWS 58 and JWS 1354. The nitric oxide (NO) and cytokines (IL-$1{\beta}$ and TNF-${\alpha}$) were measured in C57BL/6 mouse peritoneal macrophages to determine immune enhancing effects of JWS 58 and JWS 1354. A Listeria (L.) monocytogenes challenge mice model was used to evaluate immune enhancement ability of JWS 58 and JWS 1354 in vivo. The results showed that JWS 58 and JWS 1354 increased the production of NO or cytokines by peritoneal macrophages and that oral administration of viable probiotic strains in mice elicited the immuno-modulatory effect upon L. monocytogenes challenge. JWS 1354 showed stronger immune enhancing effects than JWS 58. Collectively, this study demonstrated that Lactobacillus strain JWS 58 and JWS 1354 possess immune enhancing effect. Furthermore, two stains are expected to use feed supplement to prevent diseases by pathogenic bacteria through releasing bacteriocin and enhancing host immune responses in animal.

• 대한수의학회지
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• 제20권2호
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• pp.113-118
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• 1980

Total of 11 strains of Ldctobacillus isolated from lactobacillus-fermented milk and-beverage in March 1980 were examined for susceptibility to 8 drugs, and transferability and transfer frequency of R plasmids by conjugation. Of 11 isolates each 2 strains were classified as L. cellobiosus and L. helveticus, each 1 strain as L. plantarum, L. lactis, L. acidophilus, L. delbrueckii, L. casei subsp. casei, L. casei subsp, tolerans and L. salivarius subsp, salivarius by Bergey's manual. Resistance was the most active to na lidixic acid(NA), followed in decreasing order by chloramphenicol(CP), ampicillin(AP), kanamycin(KM) and streptomycin(SM). All of isolates were resistant to NA, each 10 strains to CP and AP, 7strains to KM and 6 strains to SM, indicating all of the isolates were resistant to two or more drugs in combination. No strain was resistant to erythromycin(EM), penicillin(PC) and tetracycline(TC). The most frequently encountered resistant patterns were CP NA AP SM KM, followed by CP NA AP KM, NA AP, CP NA, CP NA AP and CP NA AP SM in order. Transfer experiment of drug resistance showed that of 11 resistant strains, 9 strains transferred parts of their resistance to AP or AP CP or SM AP, indicating 9 strains carried R plasmids determining R(AP), R(AP CP) and R(SM AP). The conjugal frequency of R(AP) from Lactobacillus to E. coli ranged from 2.5{\times}10^{-1} to $5.6{\times}10^{-4}%$, that of R(CP) ranged from 5.0{\times}10^{-1} to 5.0{\times}10^{-3}% and that of R(SM) ranged from 6.0{\times}10^{-5} to 1.4{\times}10^{-5}%, at $37^{\circ}C$ for 18 hours of incubation.