• Title/Summary/Keyword: Lactobacillus identification

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Isolation and Identification of Acid- and Bile-Tolerant Lactobacillus salivarius subsp. salivarius from Human Faeces

  • Bae, Hyoung Churl;Choi, Seong Hyun;Nam, Myoung Soo
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.8
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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.

Characterization and Immunomodulation Activity of Lactobacillus sakei L2 and L8 Isolated from Chicken Cecum (닭의 맹장으로부터 분리한 Lactobacillus sakei L2와 L8의 특성 및 면역활성)

  • Sim, Insuk;Park, Keun-Tae;Lim, Young-Hee
    • Microbiology and Biotechnology Letters
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    • v.44 no.2
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    • pp.201-207
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    • 2016
  • The aim of this study was to investigate the potential of lactic acid bacteria (LAB) strains as probiotics. Two strains were isolated from healthy chicken cecum and their acid and bile tolerance, residual organic acids, antibacterial activity against pathogenic bacteria, and immunomodulation activity were measured. Identification of the isolated strains was performed using the API 50CHL system and phylogenetic analysis using 16S rDNA sequencing. The isolates were determined to be Lactobacillus sakei strains. The acid tolerance of strains L2 and L8 was high enough that 75% of the inoculum survived in pH 2 for 2 h. The bile tolerance of both strains was observed at a 1% Oxgall concentration in MRS broth. The production of organic acids (lactic acid and acetic acid) and pH changes during growth were monitored and the maximum concentrations were obtained after 48 h of incubation. Culture supernatants of the two LAB strains showed strong antibacterial activity against pathogenic bacteria. The heat-killed LAB cells also induced high levels of immune cell proliferation compared with the control, and stimulated IL-6 and TNF-α production in mouse macrophages. Therefore, L. sakei strains L2 and L8 can be considered suitable probiotic bacteria.

Identification of the Cell-envelope Proteinase of Lactic Acid Bacteria Isolated from Kimchi. (김치 유래 젖산균의 Cell-envelope Proteinase 존재 확인)

  • 이유진;최재연;이형주;장해춘;김정환;정대균;김영석;김소미;이종훈
    • Microbiology and Biotechnology Letters
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    • v.30 no.2
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    • pp.116-122
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    • 2002
  • The partial 165 rDNA sequences of 6 lactic acid bacterial strains isolated from Kimchi were determined. Two strains were Leuconostoc mesenteroides and the rest were incorrectly classified and turned out to be Lactobacillus. As the case of dairy lactic acid bacteria, the strains isolated from Kimchi also had cell-envelope proteinase (CEP) activity. As the result of partial CEP gene amplification with CEP-specific primers, the expected 1.2-kb amplificate was obtained not from Leu. mesenteroides but from Lactobacillus strains. The deduced amino acid sequence of PCR product amplified from the genomic DNA of Lactobacillus pentosus KFR1821 showed 95% and 92% homology with those of PrtPs from Lactococcus lactis subsp. cremoris and Lactobacillus paracasei subsp. paracasei, respectively. The PCR amplificate was used as a probe and the result of Southern hybridization illuminated the location of CEP gene in chromosomal DNA of Lb. pentosus KFR1821.

Characterization and Antibacterial Activity of Lactobacillus casei HK-9 Isolated from Korean Rice Wine, Makgeolli (막걸리에서 분리한 젖산세균인 Lactobacillus casei HK-9의 특성 및 항균 활성)

  • Baek, Hyun;Choi, Moon-Seup;Oh, Kye-Heon
    • KSBB Journal
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    • v.27 no.3
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    • pp.161-166
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    • 2012
  • The purpose of this work was to examine the antibacterial activity derived from a lactic acid bacterium isolated from korean rice wine, called makgeolli. Various physiological and biochemical properties of this strain were characterized. Both the BIOLOG system and phylogenetic analysis using 16S rRNA sequencing were utilized for identification, and the strain was designated as Lactobacillus casei HK-9, and registered in GenBank as [JQ951606]. Growth rate, production of organic acids (e.g., lactic acid and acetic acid), and pH changes during growth were monitored. The maximum concentrations of lactic acid and acetic acid were approximately 576 mM and 199 mM, respectively, and pH was changed from 7.00 to 3.74 after 72 h of incubation. HPLC was used to confirm the production of lactic acid and acetic acid. Significant antimicrobial activity of the concentrated supernatant was demonstrated against various food-poison causing bacteria (e.g., Bacillus cereus, Listeria monocytogenes, Staphylococcus aureus, methicillin-resistant Staphylococcus aureus, Escherichia coli, Salmonella enteritidis). Ethanol tolerance of L. casei HK-9 showed up to 12% of ethanol within the culture.

Lactic Acid Bacteria in Total Mixed Ration Silage Containing Soybean Curd Residue: Their Isolation, Identification and Ability to Inhibit Aerobic Deterioration

  • Li, Y.;Wang, F.;Nishino, N.
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.4
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    • pp.516-522
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    • 2016
  • We investigated the effects of the predominant lactic acid bacteria (LAB) on the fermentation characteristics and aerobic stability of total mixed ration (TMR) silage containing soybean curd residue (SC-TMR silage). The SC-TMR materials were ensiled in laboratory silos for 14 or 56 days. LAB predominant in SC-TMR silage were identified (Exp. 1). Lactobacillus fermentum (L. fermentum) and Streptococcus bovis (S. bovis) were found in the untreated materials, Leuconostoc pseudomesenteroides (L. pseudomesenteroides) in 14-day silage and Lactobacillus plantarum (L. plantarum) in all silages. Pediococcus acidilactici (P. acidilactici), Lactobacillus paracasei (L. paracasei), and Lactobacillus brevis (L. brevis) formed more than 90% of the isolates in 56- day silage. Italian ryegrass and whole crop maize were inoculated with P. acidilactici and L. brevis isolates and the fermentation and aerobic stability determined (Exp. 2). Inoculation with P. acidilactici and L. brevis alone or combined improved the fermentation products in ryegrass silage and markedly enhanced its aerobic stability. In maize silage, P. acidilactici and L. brevis inoculation caused no changes and suppressed deterioration when combined with increases in acetic acid content. The results indicate that P. acidilactici and L. brevis may produce a synergistic effect to inhibit SC-TMR silage deterioration. Further studies are needed to identify the inhibitory substances, which may be useful for developing potential antifungal agents.

Isolation and Identification of Lactobacillus plantarum CIB 001 with Bile Salt Deconjugation Activity from Kimchi (김치로부터 담즙산 분해능이 우수한 Lactobacillus plantarum CIB 001의 분리 및 동정)

  • Cha, Sang-Do;Kim, Tae-Woon;Lee, Dong-Hee
    • Microbiology and Biotechnology Letters
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    • v.38 no.2
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    • pp.222-226
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    • 2010
  • This study was carried out to isolate and characterize the Lactobacillus plantarum with bile salt deconjugation activity that was isolated from Kimchi. Some isolates were selected and identified as L. plantarum by 16S rRNA gene sequence and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of whole cell protein patterns. They were assayed to determine their capacities to express bile salt hydrolase (BSH) activity. Among the identified strains, L. plantarum CIB 001 showed the highest level of BSH activity. Then, resistance to gastric acidity and bile condition were analyzed for further characterization. This strain was able to maintain viability for 1h at pH 2.0 and to survive in a MRS (deMan, Rogosa, and Sharpe) broth with 1.0% of bile acids. L. plantarum CIB 001 would potentially be useful in the food industry as probiotics.

Heat-Killed Lactobacillus plantarum KCTC 13314BP Enhances Phagocytic Activity and Immunomodulatory Effects via Activation of MAPK and STAT3 Pathways

  • Jeong, Minju;Kim, Jae Hwan;Yang, Hee;Kang, Shin Dal;Song, Seongbong;Lee, Deukbuhm;Lee, Ji Su;Park, Jung Han Yoon;Byun, Sanguine;Lee, Ki Won
    • Journal of Microbiology and Biotechnology
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    • v.29 no.8
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    • pp.1248-1254
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    • 2019
  • Identification of novel probiotic strains is of great interest in the field of functional foods. Specific strains of heat-killed bacteria have been reported to exert immunomodulatory effects. Herein, we investigated the immune-stimulatory function of heat-killed Lactobacillus plantarum KCTC 13314BP (LBP). Treatment with LBP significantly increased the production of $TNF-{\alpha}$ and IL-6 by macrophages. More importantly, LBP was able to enhance the phagocytic activity of macrophages against bacterial particles. Activation of p38, JNK, ERK, $NF-{\kappa}B$, and STAT3 was involved in the immunomodulatory function of LBP. LBP treatment significantly increased production of $TNF-{\alpha}$ by bone marrow-derived macrophages and splenocytes, further confirming the immunostimulatory effect of LBP in primary immune cells. Interestingly, the immunomodulatory effects of LBP were much stronger than those of Lactobacillus rhamnosus GG, a well-known probiotic strain. These results indicate that LBP can be a promising immune-enhancing functional food agent.

Protective Activities of Lactobacillus casei YIT 9018 against Salmonella enteritidis KU101 and Characteristics of 16S-23S rRNA Intergenic Spacer Region Sequence (Lactobacillus spp의 Salmonella enteritidis KU 101에 대한 보호 효과와 L. casei YIT 9018의 16S-23S rRNA Intergenic Spacer Region 염기배열 특성)

  • Sung, Bae-Jin;Ho, Yoon-Yung
    • Journal of Animal Science and Technology
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    • v.45 no.3
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    • pp.473-482
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    • 2003
  • In vivo protective and in vitro inhibitory activities of Lactobacillus casei YIT 9018. against typical enteritis causing Salmonella enteritidis KU101 and IgA level after challenge have been determined. In order to identify the strains of lactobacilli the sequences of 16S-23S rRNA intergenic spacer region were determined. All the test strains of Lactobacillus spp. inhibited Salmonella enteritidis, the intensity varied depending upon the species of lactobacilli. Effects on the survival rate of the mouse after challenge with Salmonella enteritidis KU101 on feeding Lactobacillus spp. have shown the highest survival rate in L. helveticus CU 631 followed by L. casei YIT 9018 and L. johnsonii C-4 and the lowest in control mice. The higher level of total Ig A concentration in the intestinal fluid of lactobacilli fed mice than control mice was observed. The sequences of 16S-23S rRNA intergenic spacer region of seven strains of Lactobacillus casei could be utilized as a strain identification, those sequences showed some degree of difference in homology.

Identification and inhibiting effect of Lactobacillus salivarius the formation of plaque and the production of volatile sulfur compounds by anaerobic bacteria (치태형성과 혐기성 세균의 황화합물 생성에 대한 Lactobacillus salivarius의 억제효과 및 동정)

  • Kim, Mi-Hyung;Sun, Gem-Ju;Ahn, Yeon-Jun
    • Journal of Korean society of Dental Hygiene
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    • v.5 no.2
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    • pp.131-145
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    • 2005
  • There are normal inhabitants doing medically useful functions in the body. There are many kinds of bacteria performing specific functions in the oral cavity. Two strains of lactic add bacteria were isolated from normal inhabitants of children's oral cavity, which inhibited the production of volatile sulfur compounds by anaerobic bacteria. The authors identified the isolates by 16S rDNA partial sequencing. 1. Two isolates were Gram-positive bacilli and produced hydrogen peroxide. 2. When Streptococcus mutans was cultured in the media, the mean weight of formed artificial plaque on the orthodontic wires was $124.4{\pm}30.4$ mg, whereas being reduced to $5.2{\pm}2.0$ mg and $10.6{\pm}6.6$ mg in the media cultured with Streptococcus mutans and each isolate, respectively(p<0.05). 3. The number of viable cells of Streptococcus mutans was $3.4{\times}10^9$ per ml in the cultured solution, whereas those of Streptococcus mutans in the combined culture with each of isolates were $4.6{\times}10^8$ and $2.4{\times}10^8$ per ml. 4. The optical density was 1.286 in the supernatant of Fusoacterium nucleatum after vortexing for 30 minutes, whereas in the supernatant of combined Fusoacterium nucleatum and each isolate, they were reduced to 0.628 and 00497, which the percentages of coaggregation between them were 2904% and 57.8%, respectively. 5. The optical density of Fusoacterium nucleatum precipitate was 1.794 in the culture media containing cysteine and $FeSO_4$ being reduced to 1.144 and 0.915 in the coaggregated precipitates of Fusoacterium nucleatum and each isolate. 6. The similarity values of 16S rDNA sequence between each of isolates and Lactobacillus salivarius subsp. salicinius were 99.60% and 99.73%, respectively, meaning that isolates were Lactobacillus salivarius subsp. salicinius. These results indicated that two strains isolated from children's saliva, which inhibited the formation of plaque and the production of volatile sulfur compounds, were identified as Lactobacillus salivarius subsp. salicinius.

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Identification of Lactobacillus ruminus SPM0211 Isolated from Healthy Koreans and Its Antimicrobial Activity against Some Pathogens

  • Yun Ji-Hee;Yim Dong-sool;Kang Jin-Yang;Kang Byung-Yong;Shin Eun-ah;Chung Myung-Jun;Kim Soo-Dong;Baek Dae-Heoun;Kim Kyungjae;Ha Nam-Joo
    • Archives of Pharmacal Research
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    • v.28 no.6
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    • pp.660-666
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    • 2005
  • The intestinal microbiota are important to the host with regard to resistance they impart against bacterial infections and their involvement in mediating metabolic functions. Lactic acid producing bacteria such as Lactobacillus play an important physiological role in these matters. The aim of the present study was to isolate Lactobacillus sp. that inhibits enteric pathogens. Initially, 17 isolates from healthy Koreans were collected on Lactobacillus selective medium. Resistance of the isolates to antibiotics including rifampicin, streptomycin, clindamycin and vancomycin was measured. One of the isolate was identified as Lactobacillus ruminus on the basis of bacterial cell morphology, cultural characteristic and biochemical characteristics, 16S rRNA sequence analysis and PCR-RAPD. Antimicrobial activity of the bacterium against Vancomycin Intermediate Resistant Staphylococcus aureus (VISA) and Vancomycin-Resistant Enterococci (VRE) was measured. About $10^4$ cells of VISA or VRE were mixed with 1, 5, and 9 mL of L. ruminus SPM 0211 and the final volume was adjusted to 10 mL with brain heart infusion (BHI) broth. The cell suspension was incubated for 3, 6, 9, and 24 h, serially diluted and then plated on BHI agar plates. As numbers of L. ruminus SPM 0211 were increased, viable cell count of VISA and VRE decreased. The strongest antimicrobial activity of SPM 0211 was observed after 9 h incubation in any mixture, almost completely inhibiting the growth of these two bacteria. The results suggest that the freshly isolated L. ruminus SPM 0211 may be used as a pro-biotic microbe that prevents the colonization of enteric pathogens and can thereby promote good gastrointestinal health.