• Title/Summary/Keyword: Lactobacillus Reuteri

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Analysis of nucleotide sequence of a novel plasmid, pILR091, from Lactobacillus reuteri L09 isolated from pig

  • Lee, Deog-Yong;Kang, Sang-Gyun;Rayamajhi, Nabin;Kang, Milan;Yoo, Han Sang
    • Korean Journal of Veterinary Research
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    • v.48 no.4
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    • pp.441-449
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    • 2008
  • The genus Lactobacillus is the largest of the genera included in lactic acid bacteria and is associated with mucosal membranes of human and animal. Only a few Lactobacillus plasmid-encoded functions have been discovered and used. In this study, a novel plasmid (pILR091) was isolated from a wild L. reuteri isolated from pig and described the characteristics of its replicons, genetic organization, and relationship with other plasmids. After digestion of the plasmid, pILR091, with SalI, plasmid DNA was cloned into the pQE-30Xa vector and sequenced. The complete sequence was confirmed by the sequencing of PCR products and analyzed with the Genbank database. The isolate copy number and stability were determined by quantitative-PCR. The complete sequence of L. reuteri contained 7,185 nucleotides with 39% G-C content and one cut site by two enzymes, SalI and HindIII. The similar ori sequence of the pC194- rolling circle replication family (TTTATATTGAT) was located 63 bp upstream of the protein replication sequence, ORF 1. Total of five ORFs was identified and the coding sequence represented 4,966 nucleotides (70.4%). ORF1 of pILR091 had a low similarity with the sequence of pTE44. Other ORFs also showed low homology and E-values. The average G-C content of pILR091 was 39%, similar with that of genomic DNA. The copy number of pILR091 was determined at approximately 24 to 25 molecules per genomic DNA. These results suggested that pILR091 might be a good candidate to construct a new vector, which could be used for cloning and expression of foreign genes in lactobacilli.

Analysis of Vaginal Lactic Acid Producing Bacteria in Healthy Women

  • Nam, Hye-Ran;Whang, Kyung-Hee;Lee, Yeon-Hee
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.515-520
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    • 2007
  • Vaginal lactic acid-producing bacteria of 80 pre-menopausal women were studied by isolation on Blood and DeMan-Rogosa-Sharpe agar, PCR with group-specific primers for Lactobacillus-denaturing gradient gel electrophoresis (DGGE), and PCR with specific primers for V3 region in 16S rRNA-temporal temperature gel electrophoresis (TTGE). Conventional isolation method on media detected only one lactobacillus (Lactobacillus brevis) while TTGE detected only Lactobacillus sp. DGGE detected seven Lactobacillus species; L. coleohominis, L. crispatus, L. iners, L. reuteri, L. rhamnosus, L. vaginalis, and Leuconostoc lactis. L. acidophilus and L. gasseri, which are prevalent in Western women, were not detected in Korean women. Furthermore, L. rhamnosus, Leuc. lactis, L. coleohominis, and Weissella cibaria, which were not previously reported in the vaginal microbiota of Korean women, were detected. The five most prevalent LABs in vaginal microbiota in Korean women were L. iners, Enterococcus faecalis, L. crispatus, Leuc. lactis, and W. cibaria.

Bile Salt Deconjugation Activity of Lactobacillus Strains Isolated from Yogurt Products (요구르트에서 분리한 Lactobacillus들의 담즙산염 분해 능력)

  • 김근배;이재환;임광세;허철성;배형석;백영진;김현욱
    • Microbiology and Biotechnology Letters
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    • v.27 no.5
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    • pp.385-390
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    • 1999
  • To investigate bile salt hydrolase activities of the bacterial strains isolated from fermented milk products, 21 strains of Lactobacillus were tested for their abilities to produced cholic acid from taurocholic and glycocholic acids. The production of cholic acid was measured by HPLC analysis during the growth in broth media for 24hrs. All strains of Lactobacillus acidophilus and L. plantarum deconjugated both taurocholate and glycocholate, whereas none strains of L. delbrueckii subsp. bulgaricus, L. casei subsp. casei, L. casei subsp. rhamnosus, L, reuteri did. L. acidophilus stains isolated from yogurts had the higher decojugation activities on glycocholate than taurocholate, however, L. acidophilus 1009 isolated from the human intestine showed the similar deconjugation activities on both taurocholate and glycocholate.

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Determination of Optimal Electrotransformation Conditions for Various Lactobacillus spp. (다양한 Lactobacillus 균주에 대한 electrotransformation 최적 조건 탐색)

  • Lee, Yoo-Won;Im, Sung-Hoon;Xin, Chun-Feng;So, Jae-Seong
    • KSBB Journal
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    • v.24 no.2
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    • pp.182-188
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    • 2009
  • Lactobacillus spp., primary members of probiotics, have significant benefits for health and well-being of human. In this study Lactobacillus strains representing six species (L. paracasei KLB58, L. fermentum MS79 and KLB282, L. plantarum KLB213, L. gasseri KLB238, and L. reuteri KLB270) isolated from Korean adults were electrotransformed with plasmid pNCKH104. To determine optimal electrotransformation conditions, various conditions including cell wall weakening agent, electroporation buffer, electric field strength and time constant were tested for each strain. Overall, high transformation efficiency of approximately 2.5 ${\times}$ $10^3$ ${\sim}$ 5.5 ${\times}$ $10^4$ CFU/${\mu}g$ DNA was obtained where conditions of 0.5 M sucrose electroporation buffer, 1.8 kV pulse voltage and 5 ms time constant were applied. The common conditions developed in this study will make transformation of various Lactobacillus spp. easier than previous procedures.

Characterization of Lactobacillus reuteri BCLR-42 and Lactobacillus plantarum BCLP-51 as novel dog probiotics with innate immune enhancing properties

  • Kim, Eun Jin;Kang, Yeong Im;Bang, Tae Il;Lee, Myoung Han;Lee, Sang Won;Choi, In Soo;Song, Chang Seon;Lee, Joong Bok;Park, Seung Yong
    • Korean Journal of Veterinary Research
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    • v.56 no.2
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    • pp.75-84
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    • 2016
  • Probiotics that are able to provide beneficial effects on animal health have become important ingredients of dog foods. This study was conducted to characterize the probiotic potentials of two strains, Lactobacillus reuteri BCLR-42 and Lactobacillus plantarum BCLP-51, that were derived from feces of healthy dogs and evaluated based on tolerance to low pH and bile acid, antimicrobial activities, enzyme profiles, sensitivity to antibiotics, and innate immune enhancing potentials. Both strains showed survival of more than 90% at pH 3 and 0.2% bile acid and exhibited broad antimicrobial activities against indicator bacteria. Moreover, both strains showed high sensitivity to antibiotics, except vancomycin, metronidazole, and gentamicin. The alkaline phosphatase was negligible (score 0), whereas they showed strong beta galactosidase activity (score range 5 or 3, respectively). The phagocytosis and oxidative burst activities of canine granulocytes were significantly enhanced in response to both strains. These results show that both strains have the capability to act as probiotics and the potential for application as ingredients in dog foods.

Inhibitory Activities of Digestive Enzymes and Antioxidant Activities of Fermented Beverages Using Momordica charantia L. (여주를 첨가한 발효음료의 소화효소 억제와 항산화 활성)

  • Park, Suin;Yeo, Seoungsoon;Lee, Youngseung;Jeong, Yoonhwa;Kim, Misook
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.11
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    • pp.1308-1315
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    • 2017
  • This study was conducted to develop Momordica charantia L. juice fermented by four Lactobacillus species such as Lactobacillus paracasei (LPA), Lactobacillus plantarum (LPL), Lactobacillus rhamnosus (LRH), and Lactobacillus reuteri (LRE) as well as to investigate their inhibitory effects against digestive enzymes and antioxidant activities. Fermentation was performed at $37^{\circ}C$ without nutrient supplementation for 72 h. The pH and total lactic acid contents were within the ranges of 3.75~3.96 and 5.21~10.04% in fermented juices, respectively. The type of starter culture and fermentation time induced changes in flavonoid contents more than total phenolic contents. All juices fermented for 48 h strongly inhibited ${\alpha}$-glucosidase activity with the percentage of inhibition ranging of 91.24~95.05%. Antioxidant activities of all juices mostly increased after 48 h of fermentation. Our results suggest that fermented juice possesses inhibitory activity against digestive enzymes and antioxidant activity, and they can be used as health functional beverages.

In vitro safety and efficacy of probiotics mixture on carbohydrate digestion inhibition (프로바이오틱스 혼합물의 in vitro에서의 안전성 및 탄수화물 소화 저해 효능 평가)

  • Eunsol Seo;Jang-Bin Woo;MinYeong Seo;Jeongmin Woo
    • Food Science and Preservation
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    • v.30 no.3
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    • pp.538-545
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    • 2023
  • This study aimed to assess the in vitro safety of a probiotics mixture (Lactobacillus acidophilus PBS066, Lactiplantibacillus plantarum PBS067, and Limosilactobacillus reuteri PBS072), along with its inhibitory effect on carbohydrate digestion. All three strains met the antibiotic resistance profile of the European Food Safety Authority (EFSA) guidelines. None of the strains exhibited hemolytic activity or cytotoxicity against Caco-2 cells. Strains PBS067 and PBS072 inhibited α-amylase activity, whereas all three strains suppressed α-glucosidase activity, indicating that the mixture might limit carbohydrate digestion in the gastrointestinal tract. These findings support the safety of this probiotics mixture and its potential to modulate carbohydrate metabolism in the gut.

Selection and immunomodulatory evaluation of lactic acid bacteria suitable for use as canine probiotics (개 생균제 사용에 적합한 유산균주의 선발 및 면역활성 평가)

  • Park, Su-Min;Park, Ho-Eun;Lee, Wan-Kyu
    • Korean Journal of Veterinary Research
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    • v.55 no.2
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    • pp.81-88
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    • 2015
  • This study was conducted to isolate lactic acid bacteria (LAB) from dog intestine and identify potential probiotic strains for canine use. One hundred and one LAB were isolated from feces of 20 healthy dogs. Acid, bile, and heat resistance along with adherence to Caco-2 cells and antimicrobial activity against pathogens were examined. To analyze immunomodulative effects, the production of nitric oxide (NO), TNF-${\alpha}$, and IL-$1{\beta}$ was measured using RAW 264.7 macrophages. Additionally, RAW BLUE cells were used to evaluate nuclear factor-${\kappa}B$ (NF-${\kappa}B$) generation. Ultimately, three strains were selected as canine probiotics and identified as Lactobacillus reuteri L10, Enterococcus faecium S33, and Bifidobacterium longum B3 by 16S rRNA sequence analysis. The L10 and S33 strains showed tolerance to pH 2.5 for 2 h, 1.0% Oxgall for 2 h, and $60^{\circ}C$ for 5 min. These strains also had strong antimicrobial activity against Escherichia coli KCTC 1682, Salmonella Enteritidis KCCM 12021, Staphylococcus aureus KCTC 1621, and Listeria monocytogenes KCTC 3569. All three strains exerted better immunomodulatory effects than Lactobacillus rhamnosus GG (LGG), a well-known commercial immunomodulatory strain, based on NO, NF-${\kappa}B$, IL-$1{\beta}$, and TNF-${\alpha}$ production. These results suggested that the three selected strains could serve as canine probiotics.

A Comparison of Feeding Lactobacillus and Virginiamycin Influence on Performance and Intestinal Microflora of Broiler Chicks (유산균과 버지니아마이신의 급여가 육계의 생산성 및 장내 미생물에 미치는 영향)

  • 김상호;박수영;유동조;이상진;류경선
    • Korean Journal of Poultry Science
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    • v.28 no.1
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    • pp.15-25
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    • 2001
  • This experiment was conducted to investigate the effect of feeding two strains of Lactobacillus and virginiamycin on performance, nutrients digestibility and intestinal microflora of broiler chicks(Abor acres$\times$Abor Acres) were randomly allocated into six treatments with four replications for five weeks. Control(no supplement), 0.05% virginiamycin(VM), Lactobacillus crispatus avibro1(LC), Lactobacillus reuteri avibro2(LR), LC+0.05% VM(LC+VM), LR+0.05% VM(LR+VM) were supplemented into basal diets, which contained ME 3,100kcal/kg and CP 22.0, 20.0% for starting and finishing period, respectively. Weight gain, feed intake and feed conversion(FC) were weekly measured. Nutrients digestibility, intestinal microflora and fecal noxious gas were examined at the end of experiment. Weight gains of chicks fed Lactobacillus or VM was significantly higher than control(P〈0.05). Feed intake increased significantly in those supplemental groups(P〈0.05). FC of chicks fed Lactobacillus or VM significantly lower than control(P〈0.05). Degestibility of crude protein, calcium, and phosphorus improved significantly in alone or combined Lactobacillus treatments(P〈0.05). Whereas DM, crude fat and ash digestibility were not statistically different. Feeding Lactobacilli tended to increase the total Lactobacillus spp. in ileum at one and three weeks of age(WOA) and showed significantly higher in cecum than control at 5 WOA. Total yeast were not shown difference at 1 and 3 WOA, but significantly increased at 5 WOA(P〈0.05). The ileal and cecal anaerobes were started to increase from the first WOA. Fecal NH$_3$gas tended to decrease in Lactobacillus treatments compared to that of other treatments.

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Effects of Dietary Supplementation of Lactobacillus on Performance, Nutrient Digestibility, Intestinal Microflora, and Fecal $NH_{3}$ Emission in Laying Hens (산란계의 생산성, 영양소 소화율 분의 암모니아 발생량 및 장내 미생물 변화에 대한 유산균의 급여 효과)

  • 김상호;유동조;박수영;이상진;최철환;나재천;류경선
    • Korean Journal of Poultry Science
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    • v.29 no.3
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    • pp.213-223
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    • 2002
  • This study was conducted to investigate the influence of feeding various Lactobacillus on production performance, nutrients digestibility, intestinal microflora, and fecal $NH_{3}$ gas emission in laying hens. Three hundred and sixty ISA Brown layers, 21 weeks of age, were randomly allotted to nine treatments, with low replicates per treatment. Nine treatments consisted of Control(no Lactobacillus), Lactobacillus crispatus avibrol(LCB: KFCC-11195), Lactobaciilus reuteri avibro2(LRB: KFCC-11196), Lactobacillus crispatus avihenl(LCH: KFCC-11197), Lactobacillus vaginalis avihen2(LVH: KFCC-11198). Each Lactobacillus was added at two levels ($10^{4}$and $10^{7}$ cfu/g diets). Egg production, and egg weight were measured daily. A metabolism trial was conducted following the 12-week feeding trial, during which egg qualities, intestinal microflora and fecal $NH_{3}$ gas emission were examined. Egg production and daily egg mass improved significantly by the addition of various Lactobacillus(P<0.05), of which effect was more notable during the latter part of the feeding trial. But, no significant differences were found among Lactobacillus strains and between two levels of supplementation. Egg weight and feed intake showed no difference among all treatments. Feed conversion ratio of birds fed lactobacillus was significantly improved compared to that of the Control(P<0.05), but not different among lactobacillus treatments. Digestibility of crude protein, ether extract and crude ash improved significantly in lactobacillus treatments(P<0.05). However, there were not statistically different by adding levels and strains. Total counts of Lactobacillus spp. in ileum of layers fed Lactobacillus were significantly higher than that of the control, but no consistent trend was found in cecum. There were no significant differences in intestinal yeast and anaerobes counts among all treatments. The Lactobaciilus supplementation did not exert my effect on the eggshell quality and Haugh unit. Fecal $NH_{3}$ gas emission decreased significantly in Lactobacillus treatments, and showed no difference between the two supplementation levels. From the result of this study, it could be concluded that dietary supplementation of Lactobacillus, regardless of their species, Improves the laying performance and decreases the fecal ammonia gas emission. The proper level of supplementation appears to be $10^{4}$ cfu/g of diet.