• 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.

• 한국식품영양과학회지
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• 제28권4호
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• pp.830-836
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• 1999

돼지장에 존재하는 많은 균총들로부터 생균제를 분리하기 위하여 생균제로서 필수 조건인 내산성, 내담즙산, 유해균 억제능력들을 조사하였는데 그 결과 다른 균들에 비해 생균제로서 작용 능력이 뛰어난 한 균주인 PA10을 얻을 수 있었다. 생균제로서 중요한 기능인 설사 유발 대장균과 Salmonella를 억제하기 위하여 돼지의 장으로부터 내산, 내담즙의 특성을 가진 미생물틀을 분리 검색하였다. 이들 중, 위산과 담즙에 잘 견디며, 대장균 및 Salmonella에 대한 억제 능력이 뛰어난 한 미생물을 분리하였는데 Lactobacillus salivarious로서 동정되었다. 이미 생물은 pH 3의 위산과 0.3% 담즙산에서 각각 2시간, 24시간 동안 약 50~70%의 생존률을 나타냈으며, 대장균 또는 Salmonella를 MRS배지에서 혼합 배양시 24시간안에 이들 유해균을 완전히 제거할 수 있었다. 이 실험에서 L salivarius로 명명된 이 균은 돼지장에 서식하는 수 많은 미생물중 내산, 내담즙산, 유해균 억제능을 동시에 만족 시킬 수 있는 특성을 가지는 균주임을 알 수 있었다. 앞으로 균주 보존 조건을 확립하고, 동물 실험에서도 유사한 결과를 얻을 수 있다면 이 분리균은 생균제로서 유용하게 이용될 수 있을 것으로 생각된다.

• 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.

• 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.

• 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.

• 한국식품과학회지
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• 제27권1호
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• pp.101-104
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• 1995

유제품 30여종과 야채발효 제품 2종에서 Lactobacillus casei ssp.(5 strains), Lactobacillus acidophilus(2 strains), Lactobacillus delbrueckii ssp. bulgaricus(6 strains), Lactobacillus plantarum(3 strains), Streptococcus salivarius ssp. thermophilus(11 strains), and Streptococcus faecalis(2 strains)를 각각 분리하여 이들의 내산성과 담즙산 내성을 평가하였다. 초기생균수를 고려하여 공분산분석으로 내산성을 비교한 결과, L. casei에 속하는 균주들의 내산성이 가장 우수하였으며, L. bulgaricus와 균주들은 가장 낮은 내산성을 보였다(p<0.05). 담즙산 내성은 L. casei, L. plantarum, L. acidophilus, S. feacalis 균주들의 경우 비슷한 경향을 보였으나, L. bulgaridus와 S. thermophilus 그룹에 속하는 균주들의 담즙산 내성은 거의 없는 것으로 나타났다.

• 대한치과의사협회지
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• 제9권12호
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• pp.807-811
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• 1971

This report was concerned with the isolation and identification of bacterial flora in the human dental plaque and the dextransucrase activity of isolated species. The results were obtained as follows: 1. The bacterial flora, isolated from the human dental plaque, was identified as 3 species of resembling streptococci, Streptococcus salivarius strain SD-1, Streptococcus bacilli, Lactobacillus brevis strain SD-3, Lactobacillus acidophilus strain SD-2 and SD-7, resembling Staphylococcus sp, and one species of resembling Leuconostoc mesenteroides strain SD-6. 2. The dextransucrase activites of resembling Streptococcus mitis strain SD-9 and Streptococcus salivarius strain SD-1 were exhibited the highest among the isolated species of human dental plaque.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• 제18권3호
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• pp.370-374
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• 2005

Two experiments were conducted to evaluate the effects of direct-fed microbial supplementation on the growth performance and nutrient digestibility in pigs. In experiment 1, forty eight pigs were used in a 42-d growth assay. There were four pigs per pen and three pens per treatment. Dietary treatments included 1) NC (without antibiotic basal diet), 2) PC (NC diet+0.1% antibiotic, 100 g/kg chlortetracycline), 3) DFM-1 (NC diet+0.2% Lactobacillus reuteri and Lactobacillus salivarius complex) and 4) DFM-2 (NC diet+0.2% Lactobacillus reuteri, Lactobacillus salivarius, Lactobacillus plantarum and Yeast complex). For the overall period, no treatment had significant effects in growth performance. In the nutrition digestibility, the pigs fed DFM diets were improved in DM and N digestibility compared with the pigs fed NC and PC diets but it was not significantly different. In experiment 2, sixty four crossbred pigs were used in a 98-d growth assay. There were four pigs per pen and four pens per treatment. Dietary treatments included 1) HND (high nutrient diet), 2) LND (low nutrient diet), 3) HND+DFM (HND diet+0.2% Lactobacillus reuteri and Lactobacillus plantarum complex) and 4) LND+DFM (LND diet+0.2% Lactobacillus reuteri and Lactobacillus plantarum complex). For overall period of growing phase, the pigs fed LND diets had improved gain/feed (p=0.01) and for overall period in the finishing phase, the pigs fed LND with DFM diets had higher ADG, ADFI and gain/feed than the others but there were no significant differences (p>0.13). In total period of growing-finishing phase, the pigs fed LND diet had higher gain/feed than the pigs fed HND diets (p<0.05). In growing phase, there were not significant differences among the treatments means for DM and N digestibility. However, the pigs fed diets with DFM had improved N digestibility (p<0.02) compared to the pigs fed diets without DFM in finishing phase. In conclusion, DFM slightly improved the growth performance in growing-finishing pigs.

• 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.