Science.gov

Sample records for lactobacillus coryniformis cect5711

  1. Lactobacillus

    MedlinePLUS

    Acidophilus, Acidophilus Bifidus, Acidophilus Lactobacillus, L. Acidophilus, L. Amylovorus, L. Brevis, L. Bulgaricus, L. Casei Immunitas, L. Casei, L. Crispatus, L. Delbrueckii, L. Fermentum, L. Gallinarum, L. ...

  2. Lactobacillus

    MedlinePLUS

    ... and also for other types of infections including urinary tract infections (UTIs), vaginal yeast infections, to prevent the common ... use lactobacillus suppositories to treat vaginal infections and urinary tract infections (UTIs). There are concerns about the quality of ...

  3. Lactobacillus apinorum sp. nov., Lactobacillus mellifer sp. nov., Lactobacillus mellis sp. nov., Lactobacillus melliventris sp. nov., Lactobacillus kimbladii sp. nov., Lactobacillus helsingborgensis sp. nov. and Lactobacillus kullabergensis sp. nov., isolated from the honey stomach of the honeybee Apis mellifera

    PubMed Central

    Alsterfjord, Magnus; Nilson, Bo; Butler, Èile; Vásquez, Alejandra

    2014-01-01

    We previously discovered a symbiotic lactic acid bacterial (LAB) microbiota in the honey stomach of the honeybee Apis mellifera. The microbiota was composed of several phylotypes of Bifidobacterium and Lactobacillus. 16S rRNA gene sequence analyses and phenotypic and genetic characteristics revealed that the phylotypes isolated represent seven novel species. One grouped with Lactobacillus kunkeei and the others belong to the Lactobacillus buchneri and Lactobacillus delbrueckiisubgroups of Lactobacillus. We propose the names Lactobacillus apinorum sp. nov., Lactobacillus mellifer sp. nov., Lactobacillus mellis sp. nov., Lactobacillus melliventris sp. nov., Lactobacillus kimbladii sp. nov., Lactobacillus helsingborgensis sp. nov. and Lactobacillus kullabergensis sp. nov. for these novel species, with the respective type strains being Fhon13NT (?=?DSM 26257T?=?CCUG 63287T), Bin4NT (?=?DSM 26254T?=?CCUG 63291T), Hon2NT (?=?DSM 26255T?=?CCUG 63289T), Hma8NT (?=?DSM 26256T?=?CCUG 63629T), Hma2NT (?=?DSM 26263T?=?CCUG 63633T), Bma5NT (?=?DSM 26265T?=?CCUG 63301T) and Biut2NT (?=?DSM 26262T?=?CCUG 63631T). PMID:24944337

  4. Selective enumeration of Lactobacillus delbrueckii ssp. bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus, bifidobacteria, Lactobacillus casei, Lactobacillus rhamnosus, and propionibacteria.

    PubMed

    Tharmaraj, N; Shah, N P

    2003-07-01

    Nineteen bacteriological media were evaluated to assess their suitability to selectively enumerate Lactobacillus delbrueckii ssp. bulgaricus, Streptococcus thermophilus, Lactobacillus casei, Lactobacillus rhamnosus, Lactobacillus acidophilus, bifidobacteria, and propionibacteria. Bacteriological media evaluated included Streptococcus thermophilus agar, pH modified MRS agar, MRS-vancomycine agar, MRS-bile agar, MRS-NaCl agar, MRS-lithium chloride agar, MRS-NNLP (nalidixic acid, neomycin sulfate, lithium chloride and paramomycine sulfate) agar, reinforced clostridial agar, sugar-based (such as maltose, galactose, sorbitol, manitol, esculin) media, sodium lactate agar, arabinose agar, raffinose agar, xylose agar, and L. casei agar. Incubations were carried out under aerobic and anaerobic conditions at 27, 30, 37, 43, and 45 degrees C for 24, 72 h, and 7 to 9 d. S. thermophilus agar and aerobic incubation at 37 degrees C for 24 h were suitable for S. thermophilus. L. delbrueckii ssp. bulgaricus could be enumerated using MRS agar (pH 4.58 or pH 5.20) and under anaerobic incubation at 45 degrees C for 72 h. MRS-vancomycine agar and anaerobic incubation at 43 degrees C for 72 h were suitable to enumerate L. rhamnosus. MRS-vancomycine agar and anaerobic incubation at 37 degrees C for 72 h were selective for L. casei. To estimate the counts of L. casei by subtraction method, counts of L. rhamnosus on MRS-vancomycine agar at 43 degrees C for 72 h under anaerobic incubation could be subtracted from total counts of L. casei and L. rhamnosus enumerated on MRS-vancomycine agar at 37 degrees C for 72 h under anaerobic incubation. L. acidophilus could be enumerated using MRS-agar at 43 degrees C for 72 h or Basal agar-maltose agar at 43 degrees C for 72 h or BA-sorbitol agar at 37 degrees C for 72 h, under anaerobic incubation. Bifidobacteria could be enumerated on MRS-NNLP agar under anaerobic incubation at 37 degrees C for 72 h. Propionibacteria could be enumerated on sodium lactate agar under anaerobic incubation at 30 degrees C for 7 to 9 d. A subtraction method was most suitable for counting propionibacteria in the presence of other lactic acid bacteria from a product. For this method, counts of lactic bacteria at d 3 on sodium lactate agar under anaerobic incubation at 30 degrees C were subtracted from counts at d 7 of lactic bacteria and propionibacteria. PMID:12906045

  5. Pentitol metabolism in Lactobacillus casei.

    PubMed Central

    London, J; Chace, N M

    1979-01-01

    Strains of Lactobacillus casei capable of growing on either ribitol or xylitol carry out a heterolactic fermentation producing ethanol, acetate, and a mixture of D- and L-lactate. Following conversion of the pentitols to ribulose 5-phosphate or xylulose 5-phosphate via enzymatic steps unique to these organisms, the intermediate products are further metabolized by enzymes of the pentose pathway. The initial enzymes of the pathway, i.e., pentitol:phosphoenolypyruvate phosphotransferase and penititol phosphate dehydrogenase, do not appear to be stringently regulated by glucose or intermediate products of glycolysis. PMID:118163

  6. Genomic diversity of Lactobacillus salivarius.

    PubMed

    Raftis, Emma J; Salvetti, Elisa; Torriani, Sandra; Felis, Giovanna E; O'Toole, Paul W

    2011-02-01

    Strains of Lactobacillus salivarius are increasingly employed as probiotic agents for humans or animals. Despite the diversity of environmental sources from which they have been isolated, the genomic diversity of L. salivarius has been poorly characterized, and the implications of this diversity for strain selection have not been examined. To tackle this, we applied comparative genomic hybridization (CGH) and multilocus sequence typing (MLST) to 33 strains derived from humans, animals, or food. The CGH, based on total genome content, including small plasmids, identified 18 major regions of genomic variation, or hot spots for variation. Three major divisions were thus identified, with only a subset of the human isolates constituting an ecologically discernible group. Omission of the small plasmids from the CGH or analysis by MLST provided broadly concordant fine divisions and separated human-derived and animal-derived strains more clearly. The two gene clusters for exopolysaccharide (EPS) biosynthesis corresponded to regions of significant genomic diversity. The CGH-based groupings of these regions did not correlate with levels of production of bound or released EPS. Furthermore, EPS production was significantly modulated by available carbohydrate. In addition to proving difficult to predict from the gene content, EPS production levels correlated inversely with production of biofilms, a trait considered desirable in probiotic commensals. L. salivarius displays a high level of genomic diversity, and while selection of L. salivarius strains for probiotic use can be informed by CGH or MLST, it also requires pragmatic experimental validation of desired phenotypic traits. PMID:21131523

  7. Lactobacillus salivarius 1077 (NRRL B-50053) bacteriocin

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Lactobacillus salivarius 1077 (NRRL B-50053) was isolated from poultry intestinal materials after demonstrating in-vitro anti-Campylobacter jejuni activity. The isolate was then used for in-vitro fermentation. The protein content of the cell-free supernatant from the spent medium was precipitated ...

  8. Antagonistic Activity of Lactobacillus Isolates against Salmonella typhi In Vitro

    PubMed Central

    Abdel-Daim, Amira; Hassouna, Nadia; Hafez, Mohamed; Ashor, Mohamed Seif Aldeen; Aboulwafa, Mohammad M.

    2013-01-01

    Background. Enteric fever is a global health problem, and rapidly developing resistance to various drugs makes the situation more alarming. The potential use of Lactobacillus to control typhoid fever represents a promising approach, as it may exert protective actions through various mechanisms. Methods. In this study, the probiotic potential and antagonistic activities of 32 Lactobacillus isolates against Salmonella typhi were evaluated. The antimicrobial activity of cell free supernatants of Lactobacillus isolates, interference of Lactobacillus isolates with the Salmonella adherence and invasion, cytoprotective effect of Lactobacillus isolates, and possibility of concurrent use of tested Lactobacillus isolates and antibiotics were evaluated by testing their susceptibilities to antimicrobial agents, and their oxygen tolerance was also examined. Results. The results revealed that twelve Lactobacillus isolates could protect against Salmonella typhi infection through interference with both its growth and its virulence properties, such as adherence, invasion, and cytotoxicity. These Lactobacillus isolates exhibited MIC values for ciprofloxacin higher than those of Salmonella typhi and oxygen tolerance and were identified as Lactobacillus plantarum. Conclusion. The tested Lactobacillus plantarum isolates can be introduced as potential novel candidates that have to be subjected for in vivo and application studies for treatment and control of typhoid fever. PMID:24191248

  9. Recombinant lactobacillus for fermentation of xylose to lactic acid and lactate

    DOEpatents

    Picataggio, Stephen K. (Golden, CO); Zhang, Min (Lakewood, CO); Franden, Mary Ann (Littleton, CO); Mc Millan, James D. (Boulder, CO); Finkelstein, Mark (Fort Collins, CO)

    1998-01-01

    A recombinant Lactobacillus MONT4 is provided which has been genetically engineered with xylose isomerase and xylulokinase genes from Lactobacillus pentosus to impart to the Lactobacillus MONT4 the ability to ferment lignocellulosic biomass containing xylose to lactic acid.

  10. Biofilm formation by vaginal Lactobacillus in vivo.

    PubMed

    Ventolini, G; Mitchell, E; Salazar, M

    2015-05-01

    Biofilm formation by nonpathogenic bacteria is responsible for their stable maintenance in vivo ecosystems as it promotes long-term permanence on the host's vaginal mucosa. Biofilm formation by Lactobacilli has been reported in vitro but not in vivo. We hypothesize the presence of biofilm formation in vivo could be also documented by microscope photographs (MP) of wet mounts obtained from uninfected vaginal samples satisfying rigorous scientific identification criteria. We analyzed 400 MP from our database, and we were able to determine that 12 MP from 6 different patients contained clues of the formation of biofilm by Lactobacilli. The most probable lactobacillus involved is presumed to be Lactobacillus jensenii. The documentation of biofilm formation by vaginal Lactobacilli at fresh wet mount preparation is significant and has several important clinical preventive and therapeutic implications. PMID:25725906

  11. Lactobacillus mixtipabuli sp. nov. isolated from total mixed ration silage.

    PubMed

    Tohno, Masanori; Kitahara, Maki; Irisawa, Tomohiro; Ohmori, Hideyuki; Masuda, Takaharu; Ohkuma, Moriya; Tajima, Kiyoshi

    2015-06-01

    Using a polyphasic taxonomic approach, we investigated three bacterial strains - IWT30T, IWT8 and IWT75 - isolated from total mixed ration silage prepared in Hachimantai, Iwate, Japan. The isolates comprised Gram-stain positive, non-motile, non-spore-forming, catalase-negative, rod-shaped bacteria. Good growth occurred at 15-45 °C and at pH 4.0-7.5. Their major cellular fatty acids were C18:1?9c and C19:1 cyclo 9,10.The G+C content of genomic DNA of strain IWT30T was 44.6 mol%. Comparative 16S rRNA gene sequence analysis showed that these novel strains belonged to the genus Lactobacillus. These strains shared 100 % 16S rRNA gene sequence similarity and were most closely related to the type strains of Lactobacillus silagei, Lactobacillus odoratitofui, Lactobacillus similis, Lactobacillus collinoides, Lactobacillus paracollinoides and Lactobacillus kimchicus, with sequence similarity values of 99.5, 98.8, 98.7, 97.8, 97.8 and 96.8 %, respectively. The level of DNA-DNA relatedness between these strains and their closest phylogenetic neighbours was less than 30 %. On the basis of additional phylogenetic analysis of pheS and rpoA gene sequences and phenotypic and chemotaxonomic characteristics, we conclude that these three strains represent a novel species of the genus Lactobacillus, for which we propose the name Lactobacillus mixtipabuli sp. nov. The type strain is IWT30T (?= JCM 19805T = DSM 28580T). PMID:25807979

  12. Epithelial cell adhesion and gastrointestinal colonization of Lactobacillus in poultry.

    PubMed

    Spivey, Megan A; Dunn-Horrocks, Sadie L; Duong, Tri

    2014-11-01

    Administration of probiotic Lactobacillus cultures is an important alternative to the use of antibiotic growth promoters and has been demonstrated to improve animal health, growth performance, and preharvest food safety in poultry production. Whereas gastrointestinal colonization is thought to be critical to their probiotic functionality, factors important to Lactobacillus colonization in chickens are not well understood. In this study we investigate epithelial cell adhesion in vitro and colonization of Lactobacillusin vivo in broiler chickens. Adhesion of Lactobacillus cultures to epithelial cells was evaluated using the chicken LMH cell line. Lactobacillus cultures were able adhere effectively to LMH cells relative to Bacillus subtilis and Salmonella Typhimurium. Epithelial cell adhesion was similar for Lactobacillus crispatus TDCC 75, L. cristpatus TDCC 76, and Lactobacillus gallinarum TDCC 77, and all 3 were more adherent than L. gallinarum TDCC 78. However, when colonization was evaluated in the ileum and cecum of broiler chicks, L. crispatus TDCC 75 and L. gallinarum TDCC 77 were more persistent than L. crispatus TDCC 76 and L. gallinarum TDCC 78. The reduction of growth in medium supplemented with oxgal was greater for L. gallinarum TDCC 78 than L. gallinarum TDCC 77, suggesting that whereas adhesion was similar for the 2 strains, the difference in colonization between L. gallinarum strains may be due in part to their bile sensitivity. This study demonstrates that whereas adhesion to epithelial cells may be important in predicting gastrointestinal colonization, other factors including bile tolerance may also contribute to the colonization of Lactobacillus in poultry. Additionally, the chicken LMH cell line is expected to provide a platform for investigating mechanisms of Lactobacillus adhesion to epithelial tissue and evaluating the probiotic potential Lactobacillus in poultry. PMID:25239531

  13. Lactobacillus Decelerates Cervical Epithelial Cell Cycle Progression

    PubMed Central

    Vielfort, Katarina; Weyler, Linda; Söderholm, Niklas; Engelbrecht, Mattias; Löfmark, Sonja; Aro, Helena

    2013-01-01

    We investigated cell cycle progression in epithelial cervical ME-180 cells during colonization of three different Lactobacillus species utilizing live cell microscopy, bromodeoxyuridine incorporation assays, and flow cytometry. The colonization of these ME-180 cells by L. rhamnosus and L. reuteri, originating from human gastric epithelia and saliva, respectively, was shown to reduce cell cycle progression and to cause host cells to accumulate in the G1 phase of the cell cycle. The G1 phase accumulation in L. rhamnosus-colonized cells was accompanied by the up-regulation and nuclear accumulation of p21. By contrast, the vaginal isolate L. crispatus did not affect cell cycle progression. Furthermore, both the supernatants from the lactic acid-producing L. rhamnosus colonies and lactic acid added to cell culture media were able to reduce the proliferation of ME-180 cells. In this study, we reveal the diversity of the Lactobacillus species to affect host cell cycle progression and demonstrate that L. rhamnosus and L. reuteri exert anti-proliferative effects on human cervical carcinoma cells. PMID:23675492

  14. Growth Stimulation of Lactobacillus Species by Lactic Streptococci1

    PubMed Central

    Branen, A. L.; Keenan, T. W.

    1969-01-01

    Cell extracts of Streptococcus species important in cheese starters stimulated the growth of Lactobacillus species common to Cheddar cheese. All Lactobacillus strains employed, with the exception of a strain of L. casei, were significantly stimulated by a strain of S. diacetilactis. L. casei was highly stimulated by both a strain of S. lactis and a strain of S. diacetilactis. The stimulant(s) was dialyzable and was partially inactivated by heat. The stimulatory principle was active at 10 C, indicating that the stimulatory effect may be influencing the growth of lactobacilli in Cheddar cheese during curing. Viable Streptococcus cells did not inhibit the growth of Lactobacillus species. PMID:5775912

  15. A Chinese rhesus macaque (Macaca mulatta) model for vaginal Lactobacillus colonization and live microbicide development

    PubMed Central

    Yu, Rosa R.; Cheng, Andrew T.; Lagenaur, Laurel A.; Huang, Wenjun; Weiss, Deborah E.; Treece, Jim; Sanders-Beer, Brigitte E.; Hamer, Dean H.; Lee, Peter P.; Xu, Qiang; Liu, Yang

    2015-01-01

    Background We sought to establish a nonhuman primate model of vaginal Lactobacillus colonization suitable for evaluating live microbial microbicide candidates. Methods Vaginal and rectal microflora in Chinese rhesus macaques (Macaca mulatta) were analyzed, with cultivable bacteria identified by 16S rRNA gene sequencing. Live lactobacilli were intravaginally administered to evaluate bacterial colonization. Results Chinese rhesus macaques harbored abundant vaginal Lactobacillus, with Lactobacillus johnsonii as the predominant species. Like humans, most examined macaques harbored only one vaginal Lactobacillus species. Vaginal and rectal Lactobacillus isolates from the same animal exhibited different genetic and biochemical profiles. Vaginal Lactobacillus was cleared by a vaginal suppository of azithromycin, and endogenous L. johnsonii was subsequently restored by intravaginal inoculation. Importantly, prolonged colonization of a human vaginal Lactobacillus jensenii was established in these animals. Conclusions The Chinese rhesus macaque harbors vaginal Lactobacillus and is a potentially useful model to support the pre-clinical evaluation of Lactobacillus-based topical microbicides. PMID:19367737

  16. Saccharomyces cerevisiae expressing bacteriophage endolysins reduce Lactobacillus contamination during fermentation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    One of the challenges facing the fuel ethanol industry is the management of bacterial contamination during fermentation. Lactobacillus species are the predominant contaminants that decrease the profitability of biofuel production by reducing ethanol yields and causing “stuck” fermentations, which i...

  17. Viricidal Effects of Lactobacillus and Yeast Fermentation

    PubMed Central

    Gilbert, Jeannine P.; Wooley, Richard E.; Shotts, Emmett B.; Dickens, J. Andra

    1983-01-01

    The survival of selected viruses in Lactobacillus- and yeast-fermented edible waste material was studied to determine the feasibility of using this material as a livestock feed ingredient. Five viruses, including Newcastle disease virus, infectious canine hepatitis virus, a porcine picornavirus, frog virus 3, and bovine virus diarrhea, were inoculated into a mixture of ground food waste (collected from a school lunch program) containing Lactobacillus acidophilus. Mixtures were incubated at 20, 30, and 40°C for 216 h. In a second trial, four viruses, including Newcastle disease virus, infectious canine hepatitis virus, frog virus 3, and a porcine picornavirus, were inoculated into similar edible waste material containing Saccharomyces cerevisiae. Mixtures were incubated at 20 and 30°C for 216 h. Samples were obtained daily for quantitative (trial 1) and qualitative (trial 2) virus isolation. Temperature, pH, and redox potential were monitored. Controlled pH and temperature studies were also done and compared with the inactivation rates in the fermentation processes. In trial 1 (Lactobacillus fermentation), infectious canine hepatitis virus survived the entire test period in the fermentation process but was inactivated below pH 4.5 in the controlled studies. Newcastle disease virus was inactivated by day 8 in the fermentation process and appeared to be primarily heat sensitive and secondarily pH sensitive in the controlled studies. The porcine picornavirus survived the fermentation process for 8 days at 20°C but was inactivated more rapidly at 30 and 40°C. The controlled studies verified these findings. Frog virus 3 was inactivated by day 3 in the fermentation process and appeared to be sensitive to low pH in the controlled studies. Bovine virus diarrhea was rapidly inactivated in the fermentation process (less than 2 h) and was pH and temperature sensitive. In trial 2 (yeast fermentation), infectious hepatitis virus survived the entire test period in the fermentation process. Newcastle disease virus was inactivated by day 7 at 20°C and day 6 at 30°C. The porcine picornavirus was inactivated by day 7 at 30°C but survived the entire test period at 20°C. Frog virus 3 was inactivated by day 3 at 20°C and day 2 at 30°C. PMID:6414372

  18. Cholesterol Assimilation by Lactobacillus Probiotic Bacteria: An In Vitro Investigation

    PubMed Central

    Jones, Mitchell L.; Shah, Divya; Jain, Poonam; Saha, Shyamali; Prakash, Satya

    2014-01-01

    Excess cholesterol is associated with cardiovascular diseases (CVD), an important cause of mortality worldwide. Current CVD therapeutic measures, lifestyle and dietary interventions, and pharmaceutical agents for regulating cholesterol levels are inadequate. Probiotic bacteria have demonstrated potential to lower cholesterol levels by different mechanisms, including bile salt hydrolase activity, production of compounds that inhibit enzymes such as 3-hydroxy-3-methylglutaryl coenzyme A, and cholesterol assimilation. This work investigates 11 Lactobacillus strains for cholesterol assimilation. Probiotic strains for investigation were selected from the literature: Lactobacillus reuteri NCIMB 11951, L. reuteri NCIMB 701359, L. reuteri NCIMB 702655, L. reuteri NCIMB 701089, L. reuteri NCIMB 702656, Lactobacillus fermentum NCIMB 5221, L. fermentum NCIMB 8829, L. fermentum NCIMB 2797, Lactobacillus rhamnosus ATCC 53103 GG, Lactobacillus acidophilus ATCC 314, and Lactobacillus plantarum ATCC 14917. Cholesterol assimilation was investigated in culture media and under simulated intestinal conditions. The best cholesterol assimilator was L. plantarum ATCC 14917 (15.18 ± 0.55?mg/1010?cfu) in MRS broth. L. reuteri NCIMB 701089 assimilated over 67% (2254.70 ± 63.33?mg/1010?cfu) of cholesterol, the most of all the strains, under intestinal conditions. This work demonstrates that probiotic bacteria can assimilate cholesterol under intestinal conditions, with L. reuteri NCIMB 701089 showing great potential as a CVD therapeutic. PMID:25295259

  19. Antimicrobial substance from a human Lactobacillus strain.

    PubMed Central

    Silva, M; Jacobus, N V; Deneke, C; Gorbach, S L

    1987-01-01

    Lactobacillus sp. strain GG, which was isolated from the feces of a normal person, produced a substance with potent inhibitory activity against a wide range of bacterial species. It inhibited anaerobic bacteria (Clostridium spp., Bacteroides spp., Bifidobacterium spp.), members of the family Enterobacteriaceae, Pseudomonas spp. Staphylococcus spp., and Streptococcus spp., as demonstrated by a microbiological assay; however, it did not inhibit other lactobacilli. The inhibitory activity occurred between pH 3 and 5 and was heat stable. Bactericidal activity against Escherichia coli was demonstrated at a dilution of 1:128. The inhibitory substance was distinct from lactic and acetic acids. It had a low molecular weight (less than 1,000) and was soluble in acetone-water (10:1). Because of these characteristics, the inhibitory material could not be considered a bacteriocin; it most closely resembled a microcin, which has been associated previously with members of the family Enterobacteriaceae. PMID:3307619

  20. Molecular Characterization of Lactobacillus plantarum DMDL 9010, a Strain with Efficient Nitrite Degradation Capacity

    PubMed Central

    Fei, Yong-tao; Liu, Dong-mei; Luo, Tong-hui; Chen, Gu; Wu, Hui; Li, Li; Yu, Yi-gang

    2014-01-01

    Nitrites commonly found in food, especially in fermented vegetables, are potential carcinogens. Therefore, limiting nitrites in food is critically important for food safety. A Lactobacillus strain (Lactobacillus sp. DMDL 9010) was previously isolated from fermented vegetables by our group, and is not yet fully characterized. A number of phenotypical and genotypical approaches were employed to characterize Lactobacillus sp. DMDL 9010. Its nitrite degradation capacity was compared with four other Lactobacillus strains, including Lactobacillus casei subsp. rhamnosus 719, Lactobacillus delbrueckii subsp. bulgaricu 1.83, Streptococcus thermophilus 1.204, and lactobacillus plantarum 8140, on MRS medium. Compared to these four Lactobacillus strains, Lactobacillus sp. DMDL 9010 had a significantly higher nitrite degradation capacity (P<0.001). Based on 16S rDNA sequencing and sequence comparison, Lactobacillus sp. DMDL 9010 was identified as either Lactobacillus plantarum or Lactobacillus pentosus. To further identify this strain, the flanking regions (922 bp and 806 bp upstream and downstream, respectively) of the L-lactate dehydrogenase 1 (L-ldh1) gene were amplified and sequenced. Lactobacillus sp. DMDL 9010 had 98.92 and 76.98% sequence identity in the upstream region with L. plantarum WCFS1 and L. pentosus IG1, respectively, suggesting that Lactobacillu sp. DMDL 9010 is an L. plantarum strain. It was therefore named L. plantarum DMDL 9010. Our study provides a platform for genetic engineering of L. plantarum DMDL 9010, in order to further improve its nitrite degradation capacity. PMID:25423449

  1. 21 CFR 184.1924 - Urease enzyme preparation from Lactobacillus fermentum.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ...2012-04-01 2012-04-01 false Urease enzyme preparation from Lactobacillus fermentum...Affirmed as GRAS § 184.1924 Urease enzyme preparation from Lactobacillus fermentum. (a) This enzyme preparation is derived from the...

  2. 21 CFR 184.1924 - Urease enzyme preparation from Lactobacillus fermentum.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ...2014-04-01 2014-04-01 false Urease enzyme preparation from Lactobacillus fermentum...Affirmed as GRAS § 184.1924 Urease enzyme preparation from Lactobacillus fermentum. (a) This enzyme preparation is derived from the...

  3. 21 CFR 184.1924 - Urease enzyme preparation from Lactobacillus fermentum.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ...2010-04-01 2009-04-01 true Urease enzyme preparation from Lactobacillus fermentum...Affirmed as GRAS § 184.1924 Urease enzyme preparation from Lactobacillus fermentum. (a) This enzyme preparation is derived from the...

  4. 21 CFR 184.1924 - Urease enzyme preparation from Lactobacillus fermentum.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ...2011-04-01 2011-04-01 false Urease enzyme preparation from Lactobacillus fermentum...Affirmed as GRAS § 184.1924 Urease enzyme preparation from Lactobacillus fermentum. (a) This enzyme preparation is derived from the...

  5. 21 CFR 184.1924 - Urease enzyme preparation from Lactobacillus fermentum.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ...2013-04-01 2013-04-01 false Urease enzyme preparation from Lactobacillus fermentum...Affirmed as GRAS § 184.1924 Urease enzyme preparation from Lactobacillus fermentum. (a) This enzyme preparation is derived from the...

  6. Recombinant lactobacillus for fermentation of xylose to lactic acid and lactate

    DOEpatents

    Picataggio, S.K.; Zhang, M.; Franden, M.A.; McMillan, J.D.; Finkelstein, M.

    1998-08-25

    A recombinant Lactobacillus MONT4 is provided which has been genetically engineered with xylose isomerase and xylulokinase genes from Lactobacillus pentosus to impart to the Lactobacillus MONT4 the ability to ferment lignocellulosic biomass containing xylose to lactic acid. 4 figs.

  7. Enteral feeding of premature infants with Lactobacillus GG.

    PubMed Central

    Millar, M R; Bacon, C; Smith, S L; Walker, V; Hall, M A

    1993-01-01

    The objectives of this study were to determine whether or not the probiotic Lactobacillus GG can colonise the immature bowel of premature infants and if so, does colonisation result in a reduction of the size of the bowel reservoir of nosocomial pathogens such as enterobacteriaceae, enterococci, yeasts or staphylococci, and does colonisation with Lactobacillus GG have any effect on the clinical progress and outcome. Twenty preterm infants with a gestational age of 33 weeks or less who were resident on a neonatal unit were studied from the initiation of milk feeds until discharge. The infants were randomised to receive either milk feeds or milk feeds supplemented with Lactobacillus GG 10(8) colony forming units twice a day for two weeks. The clinical features of the two groups of infants were similar. Orally administered Lactobacillus GG was well tolerated and did colonise the bowel of premature infants. However, colonisation with Lactobacillus GG did not reduce the faecal reservoir of potential pathogens and there was no evidence that colonisation had any positive clinical benefit for this particular group of infants. PMID:8285750

  8. Characterization of Selected Lactobacillus Strains for Use as Probiotics

    PubMed Central

    Song, Minyu; Yun, Bohyun; Moon, Jae-Hak; Park, Dong-June; Lim, Kwangsei; Oh, Sejong

    2015-01-01

    The aim of this study was to evaluate the functional properties of lactic acid bacteria from various sources and to identify strains for use as probiotics. Ten Lactobacillus strains were selected and their properties such as bile tolerance, acid resistance, cholesterol assimilation activity, and adherence to HT-29 cells were assessed to determine their potential as probiotics. Lactobacillus sp. JNU 8829, L. casei MB3, L. sakei MA9, L. sakei CH8, and L. acidophilus M23 were found to show full tolerance to the 0.3% bile acid. All strains without L. acidophilus M23 were the most acid-tolerant strains. After incubating the strains at pH 2.5 for 2 h, their viability decreased by 3 Log cells. Some strains survived at pH 2.5 in the presence of pepsin and 0.3% bile acid. Lactobacillus sp. JNU 8829, L. acidophilus KU41, L. acidophilus M23, L. fermentum NS2, L. plantarum M13, and L. plantarum NS3 were found to reduce cholesterol levels by >50% in vitro. In the adhesion assay, Lactobacillus sp. JNU 8829, L. casei MB3, L. sakei MA9, and L. sakei CH8 showed higher adhesion activities after 2 h of co-incubation with the intestinal cells. The results of this comprehensive analysis shows that this new probiotic strain named, Lactobacillus sp. JNU 8829 could be a promising candidate for dairy products.

  9. Salivary lactobacillus counts in the prediction of caries activity.

    PubMed

    Crossner, C G

    1981-08-01

    The aim of the study was to investigate the reliability and clinical value for prediction of caries activity of determining the number of lactobacilli in saliva, the salivary secretion rate and the presence of yeasts in the saliva. For this purpose one entire age group of pupils (115 14-year-olds) attending one of the compulsory 9-year comprehensive schools in Orebro was selected for longitudinal examination. Over a period of 64 weeks three clinical recordings were made and seven salivary samples were collected. The correlations between caries activity, caries frequency (DFS), lactobacillus counts, yeasts in saliva, salivary secretion rate and gingivitis were examined. The results showed that in spite of a well-organized dental health service, including adequate preventive measures, a pronounced variation in caries activity remained and thus a means of predicting the onset of caries would be a valuable asset. Statistically significant correlations were found between caries activity on the one hand and caries frequency and lactobacillus counts on the other. The lactobacillus count was found to be a suitable measure in predicting caries activity. It was concluded that the lactobacillus count should be used for caries prediction in healthy, properly treated patients. It is important that there are no areas of microbial retention on the teeth such as open carious lesions, poorly executed restorations, dentures or orthodontic bands. In such situations, the lactobacillus count seems to reflect the frequency of ingested fermentable carbohydrates and thus, indirectly, the risk of initiating carious lesions. PMID:6949671

  10. Lactobacillus rossiae, a Vitamin B12 Producer, Represents a Metabolically Versatile Species within the Genus Lactobacillus

    PubMed Central

    De Angelis, Maria; Bottacini, Francesca; Fosso, Bruno; Kelleher, Philip; Calasso, Maria; Di Cagno, Raffaella; Ventura, Marco; Picardi, Ernesto; van Sinderen, Douwe; Gobbetti, Marco

    2014-01-01

    Lactobacillus rossiae is an obligately hetero-fermentative lactic acid bacterium, which can be isolated from a broad range of environments including sourdoughs, vegetables, fermented meat and flour, as well as the gastrointestinal tract of both humans and animals. In order to unravel distinctive genomic features of this particular species and investigate the phylogenetic positioning within the genus Lactobacillus, comparative genomics and phylogenomic approaches, followed by functional analyses were performed on L. rossiae DSM 15814T, showing how this type strain not only occupies an independent phylogenetic branch, but also possesses genomic features underscoring its biotechnological potential. This strain in fact represents one of a small number of bacteria known to encode a complete de novo biosynthetic pathway of vitamin B12 (in addition to other B vitamins such as folate and riboflavin). In addition, it possesses the capacity to utilize an extensive set of carbon sources, a characteristic that may contribute to environmental adaptation, perhaps enabling the strain's ability to populate different niches. PMID:25264826

  11. Genome Instability in Lactobacillus rhamnosus GG

    PubMed Central

    Molenaar, Douwe; van IJcken, Wilfred; Venema, Koen

    2013-01-01

    We describe here a comparative genome analysis of three dairy product isolates of Lactobacillus rhamnosus GG (LGG) and the ATCC 53103 reference strain to the published genome sequence of L. rhamnosus GG. The analysis showed that in two of three isolates, major DNA segments were missing from the genomic islands LGGISL1,2. The deleted DNA segments consist of 34 genes in one isolate and 84 genes in the other and are flanked by identical insertion elements. Among the missing genes are the spaCBA genes, which encode pilin subunits involved in adhesion to mucus and persistence of the strains in the human intestinal tract. Subsequent quantitative PCR analyses of six commercial probiotic products confirmed that two more products contain a heterogeneous population of L. rhamnosus GG variants, including genotypes with or without spaC. These results underline the relevance for quality assurance and control measures targeting genome stability in probiotic strains and justify research assessing the effect of genetic rearrangements in probiotics on the outcome of in vitro and in vivo efficacy studies. PMID:23354703

  12. Health-Promoting Properties of Lactobacillus helveticus

    PubMed Central

    Taverniti, Valentina; Guglielmetti, Simone

    2012-01-01

    Lactobacillus helveticus is an important industrial thermophilic starter that is predominantly employed in the fermentation of milk for the manufacture of several cheeses. In addition to its technological importance, a growing body of scientific evidence shows that strains belonging to the L. helveticus species have health-promoting properties. In this review, we synthesize the results of numerous primary literature papers concerning the ability of L. helveticus strains to positively influence human health. Several in vitro studies showed that L. helveticus possesses many common probiotic properties, such as the ability to survive gastrointestinal transit, adhere to epithelial cells, and antagonize pathogens. In vivo studies in murine models showed that L. helveticus could prevent gastrointestinal infections, enhance protection against pathogens, modulate host immune responses, and affect the composition of the intestinal microbiota. Interventional studies and clinical trials have also demonstrated a number of health-promoting properties of L. helveticus. Finally, several studies suggested that specific enzymatic activities of L. helveticus could indirectly benefit the human host by enhancing the bioavailability of nutrients, removing allergens and other undesired molecules from food, and producing bioactive peptides through the digestion of food proteins. In conclusion, this review demonstrates that in light of the scientific literature presented, L. helveticus can be included among the bacterial species that are generally considered to be probiotic. PMID:23181058

  13. Genome sequence and analysis of Lactobacillus helveticus

    PubMed Central

    Cremonesi, Paola; Chessa, Stefania; Castiglioni, Bianca

    2013-01-01

    The microbiological characterization of lactobacilli is historically well developed, but the genomic analysis is recent. Because of the widespread use of Lactobacillus helveticus in cheese technology, information concerning the heterogeneity in this species is accumulating rapidly. Recently, the genome of five L. helveticus strains was sequenced to completion and compared with other genomically characterized lactobacilli. The genomic analysis of the first sequenced strain, L. helveticus DPC 4571, isolated from cheese and selected for its characteristics of rapid lysis and high proteolytic activity, has revealed a plethora of genes with industrial potential including those responsible for key metabolic functions such as proteolysis, lipolysis, and cell lysis. These genes and their derived enzymes can facilitate the production of cheese and cheese derivatives with potential for use as ingredients in consumer foods. In addition, L. helveticus has the potential to produce peptides with a biological function, such as angiotensin converting enzyme (ACE) inhibitory activity, in fermented dairy products, demonstrating the therapeutic value of this species. A most intriguing feature of the genome of L. helveticus is the remarkable similarity in gene content with many intestinal lactobacilli. Comparative genomics has allowed the identification of key gene sets that facilitate a variety of lifestyles including adaptation to food matrices or the gastrointestinal tract. As genome sequence and functional genomic information continues to explode, key features of the genomes of L. helveticus strains continue to be discovered, answering many questions but also raising many new ones. PMID:23335916

  14. Biofilms of Lactobacillus plantarum and Lactobacillus fermentum: Effect on stress responses, antagonistic effects on pathogen growth and immunomodulatory properties.

    PubMed

    Aoudia, Nabil; Rieu, Aurélie; Briandet, Romain; Deschamps, Julien; Chluba, Johanna; Jego, Gaëtan; Garrido, Carmen; Guzzo, Jean

    2016-02-01

    Few studies have extensively investigated probiotic functions associated with biofilms. Here, we show that strains of Lactobacillus plantarum and Lactobacillus fermentum are able to grow as biofilm on abiotic surfaces, but the biomass density differs between strains. We performed microtiter plate biofilm assays under growth conditions mimicking to the gastrointestinal environment. Osmolarity and low concentrations of bile significantly enhanced Lactobacillus spatial organization. Two L. plantarum strains were able to form biofilms under high concentrations of bile and mucus. We used the agar well-diffusion method to show that supernatants from all Lactobacillus except the NA4 isolate produced food pathogen inhibitory molecules in biofilm. Moreover, TNF-? production by LPS-activated human monocytoid cells was suppressed by supernatants from Lactobacillus cultivated as biofilms but not by planktonic culture supernatants. However, only L. fermentum NA4 showed anti-inflammatory effects in zebrafish embryos fed with probiotic bacteria, as assessed by cytokine transcript level (TNF-?, IL-1? and IL-10). We conclude that the biofilm mode of life is associated with beneficial probiotic properties of lactobacilli, in a strain dependent manner. Those results suggest that characterization of isolate phenotype in the biofilm state could be additional valuable information for the selection of probiotic strains. PMID:26611169

  15. Nearly Complete Genome Sequence of Lactobacillus plantarum Strain NIZO2877

    PubMed Central

    Bayjanov, Jumamurat R.; Joncour, Pauline; Hughes, Sandrine; Gillet, Benjamin; Kleerebezem, Michiel; Siezen, Roland; van Hijum, Sacha A. F. T.

    2015-01-01

    Lactobacillus plantarum is a versatile bacterial species that is isolated mostly from foods. Here, we present the first genome sequence of L. plantarum strain NIZO2877 isolated from a hot dog in Vietnam. Its two contigs represent a nearly complete genome sequence. PMID:26607887

  16. Lactobacillus plantarum mediated fermentation of Psidium guajava L. fruit extract.

    PubMed

    Bhat, Ravish; Suryanarayana, Lakshminarayana Chikkanayakanahalli; Chandrashekara, Karunakara Alageri; Krishnan, Padma; Kush, Anil; Ravikumar, Puja

    2015-04-01

    Sixteen hour fermentation of the white flesh raw guava Lucknow 49 cultivar using Lactobacillus plantarum NCIM 2912 was taken up for enhancing the antioxidant potential. The fermented guava product with high antioxidant potential, total phenolic content and short and medium chain fatty acids can be used as functional food. PMID:25300190

  17. Draft Genome Sequence of Lactobacillus fermentum NB-22

    PubMed Central

    Shkoporov, A. N.; Efimov, B. A.; Pikina, A. P.; Borisova, O. Y.; Gladko, I. A.; Postnikova, E. A.; Lordkipanidze, A. E.; Kafarskaia, L. I.

    2015-01-01

    We announce here a draft genome sequence of Lactobacillus fermentum NB-22, a strain isolated from human vaginal microbiota. The assembled sequence consists of 190 contigs, joined into 137 scaffolds, and the total size is 2.01 Mb. PMID:26272572

  18. Lactobacillus kimchiensis sp. nov., isolated from a fermented food

    E-print Network

    Bae, Jin-Woo

    Lactobacillus kimchiensis sp. nov., isolated from a fermented food Jandi Kim,3 Joon Yong Kim,3 Min bacterium was isolated from a traditional fermented food, kimchi. The morphology, physiology, biochemical ). Kimchi is a traditional Korean fermented food made of various vegetables and spices (red pepper, garlic

  19. Sequence of ornithine decarboxylase from Lactobacillus sp. strain 30a.

    PubMed Central

    Hackert, M L; Carroll, D W; Davidson, L; Kim, S O; Momany, C; Vaaler, G L; Zhang, L

    1994-01-01

    A gene encoding biodegradative ornithine decarboxylase from Lactobacillus sp. strain 30a was isolated from a genomic DNA library and sequenced. Primer extension analysis revealed two transcription initiation sites. The deduced amino acid sequence is compared with the amino acid sequences of five previously reported bacterial decarboxylases, and conserved pyridoxal phosphate motif residues are identified. PMID:7961515

  20. First Functional and Mutational Analysis of Group 3 N-Acetylneuraminate Lyases from Lactobacillus antri and Lactobacillus sakei 23K

    PubMed Central

    García-García, María Inmaculada; Gil-Ortiz, Fernando; García-Carmona, Francisco; Sánchez-Ferrer, Álvaro

    2014-01-01

    N-acetyl neuraminate lyases (NALs) catalyze the reversible aldol cleavage of N-acetyl neuraminic acid (Neu5Ac) to pyruvate and N-acetyl-D-mannosamine (ManNAc). Previous phylogenetic studies divided NALs into four different groups. Groups 1 and 2 have been well characterized at both kinetic and molecular levels, but no NAL from group 3 has been studied to date. In this work, a functional characterization of two group 3 members was performed using the recombinant NALs from Lactobacillus antri and Lactobacillus sakei 23K, revealing an optimal pH of between 6.0 and 7.0, low stability at basic pHs (>8.0), low optimal temperatures and, especially, low catalytic efficiency compared with their counterparts in group 1 and 2. The mutational analysis carried out showed that a plausible molecular reason for the low activity shown by Lactobacillus antri and Lactobacillus sakei 23k NALs compared with group 1 and 2 NALs could be the relatively small sugar-binding pocket they contain. A functional divergence analysis concluding that group 3 is more closely related to group 2 than to group 1. PMID:24817128

  1. The Effects of Two Lactobacillus plantarum Strains on Rat Lipid Metabolism Receiving a High Fat Diet

    PubMed Central

    Salaj, Rastislav; Štofilová, Jana; Šoltesová, Alena; Hertelyová, Zdenka; Hijová, Emília; Bertková, Izabela; Strojný, Ladislav; Kružliak, Peter

    2013-01-01

    The aim of our study was to evaluate the effects of the different probiotic strains, Lactobacillus plantarum LS/07 and Lactobacillus plantarum Biocenol LP96, on lipid metabolism and body weight in rats fed a high fat diet. Compared with the high fat diet group, the results showed that Lactobacillus plantarum LS/07 reduced serum cholesterol and LDL cholesterol, but Lactobacillus plantarum Biocenol LP96 decreased triglycerides and VLDL, while there was no change in the serum HDL level and liver lipids. Both probiotic strains lowered total bile acids in serum. Our strains have no significant change in body weight, gain weight, and body fat. These findings indicate that the effect of lactobacilli on lipid metabolism may differ among strains and that the Lactobacillus plantarum LS/07 and Lactobacillus plantarum Biocenol LP96 can be used to improve lipid profile and can contribute to a healthier bowel microbial balance. PMID:24470789

  2. Survey of compound microsatellites in multiple Lactobacillus genomes.

    PubMed

    Basharat, Zarrin; Yasmin, Azra

    2015-12-01

    Distinct simple sequence repeats with 2 or more individual microsatellites joined together or lying adjacent to each other are identified as compound microsatellites. Investigation of such composite microsatellites in the genomes of genus Lactobacillus was the aim of this study. In silico inspection of microsatellite clustering in genomes of 14 Lactobacillus species revealed a wealth of compound microsatellites. All of the mined compound microsatellites were imperfect, were composed of variant motifs, and increased in all genomes, with maximum distance (dMAX) increments of 10 to 50. The majority of these repeats were present in the coding regions. A correlation of microsatellite to compound microsatellite density was detected. The difference established in compound microsatellite division among eukaryotes, Escherichia coli, and lactobacilli is suggestive of diverse genomic features and elementary distinction between creation and fixation methods of compound microsatellites among these organisms. PMID:26445296

  3. Probiotic properties of Lactobacillus strains isolated from Tibetan kefir grains.

    PubMed

    Zheng, Yongchen; Lu, Yingli; Wang, Jinfeng; Yang, Longfei; Pan, Chenyu; Huang, Ying

    2013-01-01

    The objective of this study was to evaluate the functional properties of lactic acid bacteria (LAB) isolated from Tibetan kefir grains. Three Lactobacillus isolates identified as Lactobacillus acidophilus LA15, Lactobacillus plantarum B23 and Lactobacillus kefiri D17 that showed resistance to acid and bile salts were selected for further evaluation of their probiotic properties. The 3 selected strains expressed high in vitro adherence to Caco-2 cells. They were sensitive to gentamicin, erythromycin and chloramphenicol and resistant to vancomycin with MIC values of 26 µg/ml. All 3 strains showed potential bile salt hydrolase (BSH) activity, cholesterol assimilation and cholesterol co-precipitation ability. Additionally, the potential effect of these strains on plasma cholesterol levels was evaluated in Sprague-Dawley (SD) rats. Rats in 4 treatment groups were fed the following experimental diets for 4 weeks: a high-cholesterol diet, a high-cholesterol diet plus LA15, a high-cholesterol diet plus B23 or a high-cholesterol diet plus D17. The total cholesterol, triglyceride and low-density lipoprotein cholesterol levels in the serum were significantly (P<0.05) decreased in the LAB-treated rats compared with rats fed a high-cholesterol diet without LAB supplementation. The high-density lipoprotein cholesterol levels in groups B23 and D17 were significantly (P<0.05) higher than those in the control and LA15 groups. Additionally, both fecal cholesterol and bile acid levels were significantly (P<0.05) increased after LAB administration. Fecal lactobacilli counts were significantly (P<0.05) higher in the LAB treatment groups than in the control groups. Furthermore, the 3 strains were detected in the rat small intestine, colon and feces during the feeding trial. The bacteria levels remained high even after the LAB administration had been stopped for 2 weeks. These results suggest that these strains may be used in the future as probiotic starter cultures for manufacturing novel fermented foods. PMID:23894554

  4. Resequencing of the Lactobacillus plantarum Strain WJL Genome

    PubMed Central

    Bayjanov, Jumamurat R.; Joncour, Pauline; Hughes, Sandrine; Gillet, Benjamin; Kleerebezem, Michiel; Siezen, Roland; van Hijum, Sacha A. F. T.

    2015-01-01

    Lactobacillus plantarum strain WJL is a symbiont isolated from the Drosophila melanogaster gut. The genome of L. plantarum WJL, first sequenced in 2013, was resequenced and rescaffolded in this study. A combination of Sanger and Illumina sequencing allowed us to reduce the number of contigs from 102 to 13. This work contributes to a better understanding of the genome and function of this organism. PMID:26607892

  5. The effect of probiotics (Lactobacillus rhamnosus HN001, Lactobacillus paracasei LPC-37, and Lactobacillus acidophilus NCFM) on the availability of minerals from Dutch-type cheese.

    PubMed

    Aljewicz, Marek; Siemianowska, Ewa; Cichosz, Gra?yna; To?ska, El?bieta

    2014-01-01

    The use of probiotic cultures in the production of Dutch-type cheeses did not lead to significant changes in their chemical composition but it lowered their acidity. The availability of calcium and magnesium analyzed by in vitro enzymatic hydrolysis was 19 and 35%, respectively; the availability of phosphorus was significantly higher, at >90%. The use of probiotic cultures significantly increased the availability of calcium (~2.5%), phosphorus (~6%), and magnesium (~18%). The in vitro method supports accurate determination of the effect of the Lactobacillus spp. cultures on the availability of mineral compounds ingested with Dutch-type cheese. PMID:24913654

  6. Development of a quantitative PCR assay for rapid detection of Lactobacillus plantarum and Lactobacillus fermentum in cocoa bean fermentation.

    PubMed

    Schwendimann, Livia; Kauf, Peter; Fieseler, Lars; Gantenbein-Demarchi, Corinne; Miescher Schwenninger, Susanne

    2015-08-01

    To monitor dominant species of lactic acid bacteria during cocoa bean fermentation, i.e. Lactobacillus plantarum and Lactobacillus fermentum, a fast and reliable culture-independent qPCR assay was developed. A modified DNA isolation procedure using a commercial kit followed by two species-specific qPCR assays resulted in 100% sensitivity for L. plantarum and L. fermentum. Kruskal-Wallis and post-hoc analyses of data obtained from experiments with cocoa beans that were artificially spiked with decimal concentrations of L. plantarum and L. fermentum strains allowed the calculation of a regression line suitable for the estimation of both species with a detection limit of 3 to 4 Log cells/g cocoa beans. This process was successfully tested for efficacy through the analyses of samples from laboratory-scale cocoa bean fermentations with both the qPCR assay and a culture-dependent method which resulted in comparable results. PMID:26026241

  7. Plant extract enhances the viability of Lactobacillus delbrueckii subsp. bulgaricus and Lactobacillus acidophilus in probiotic nonfat yogurt.

    PubMed

    Michael, Minto; Phebus, Randall K; Schmidt, Karen A

    2015-01-01

    A commercial plant extract (prepared from olive, garlic, onion and citrus extracts with sodium acetate (SA) as a carrier) was evaluated to extend the viability of yogurt starter and probiotic bacteria as a means to enhance the shelf life of live and active culture, probiotic nonfat yogurt. Yogurts prepared from three different formulas (0.5* plant extract, 0.25* SA, or no supplement) and cultures (yogurt starter plus Bifidobacterium animalis,Lactobacillus acidophilus, or both probiotics) were assessed weekly during 29 days of storage at 5°C. Supplemented yogurt mixes had greater buffering capacities than non-supplemented yogurt mixes. At the end of storage, Lactobacillus bulgaricus and L. acidophilus counts in supplemented yogurts were greater compared with non-supplemented yogurts. Supplementation did not affect Streptococcus thermophilus and B. animalis counts. Hence the greater buffering capacity of yogurt containing plant extract could enhance the longevity of the probiotics, L. bulgaricus and L. acidophilus, during storage. PMID:25650127

  8. Systemic augmentation of the immune response in mice by feeding fermented milks with Lactobacillus casei and Lactobacillus acidophilus.

    PubMed Central

    Perdigón, G; de Macias, M E; Alvarez, S; Oliver, G; de Ruiz Holgado, A P

    1988-01-01

    This study investigates the effect of feeding fermented milks with Lactobacillus casei, Lactobacillus acidophilus and a mixture of both micro-organisms on the specific and non-specific host defence mechanisms in Swiss mice. Animals fed with fermented milk for 8 days (100 micrograms/day) showed an increase in both phagocytic and lymphocytic activity. This activation of the immune system began on the 3rd day, reached a maximum on the 5th, and decreased slightly on the 8th day of feeding. In the 8-day treated mice, boosted with a single dose (100 micrograms) on the 11th day, the immune response increased further. The feeding with fermented milk produced neither hepatomegaly nor splenomegaly. These results suggest that L. casei and L. acidophilus, associated with intestinal mucosae, can influence the level of activation of the immune system. The possible clinical application of fermented milks as immunopotentiators is also discussed. PMID:3123370

  9. Synergistic impact of Lactobacillus fermentum, Lactobacillus plantarum and vincristine on 1,2-dimethylhydrazine-induced colorectal carcinogenesis in mice

    PubMed Central

    ASHA; GAYATHRI, DEVARAJA

    2012-01-01

    Lactobacillus sp. is the most dominant probiotic strain of bacteria. Evidence indicates that the consumption of Lactobacillus sp. reduces the risk of colorectal cancer in animal models. The present study was carried out to determine whether administration of Lactobacillus fermentum/ Lactobacillus plantarum alone or in combination with vincristine have a synergistic impact on the control of colorectal cancer in an animal model. Mice with 1,2 dimethylhydrazine (DMH) hydrochloride-induced colon cancer were fed with L. fermentum and L. plantarum isolated along with vincristine. An increase in body weight, a decrease in ammonia concentration, a decrease in ? glucosidase and ? glucuronidase enzyme activity and a reduction in the number of crypts in the mice in the pre-carcinogen-induced group was noted when compared to these variables in the post-carcinogen-induced group. The body weight of the mice fed L. fermentum along with vincristine was increased (6.5 g), and was found to be 3.5 times higher compared to that of the control. A marked decrease in the ammonia concentration (240 mg), and ? glucosidase (0.0023 IU) and ? glucopyranose enzyme activity (0.0027 IU) was observed; 22.59% less ammonia concentration, 73.26% less ? glucosidase activity and 56.46% less ? glucuronidase enzyme activity was noted when compared to the control. A significant reduction in the number of aberrant crypt foci (ACF) (90%) was observed when compared to the control. Maximum protection was observed in the mice fed the probiotics and vincristine prior to cancer induction. Among the different dietary combinations tested in the present study, L. fermentum and vincristine showed a more extensive reduction in ammonia concentration, ? glucosidase, ? glucuronidase activity and the number of ACF. PMID:22970015

  10. Effects of Lactobacillus johnsonii and Lactobacillus reuteri on gut barrier function and heat shock proteins in intestinal porcine epithelial cells

    PubMed Central

    Liu, Hao-Yu; Roos, Stefan; Jonsson, Hans; Ahl, David; Dicksved, Johan; Lindberg, Jan Erik; Lundh, Torbjörn

    2015-01-01

    Heat shock proteins (HSPs) are a set of highly conserved proteins that can serve as intestinal gate keepers in gut homeostasis. Here, effects of a probiotic, Lactobacillus rhamnosus GG (LGG), and two novel porcine isolates, Lactobacillus johnsonii strain P47-HY and Lactobacillus reuteri strain P43-HUV, on cytoprotective HSP expression and gut barrier function, were investigated in a porcine IPEC-J2 intestinal epithelial cell line model. The IPEC-J2 cells polarized on a permeable filter exhibited villus-like cell phenotype with development of apical microvilli. Western blot analysis detected HSP expression in IPEC-J2 and revealed that L. johnsonii and L. reuteri strains were able to significantly induce HSP27, despite high basal expression in IPEC-J2, whereas LGG did not. For HSP72, only the supernatant of L. reuteri induced the expression, which was comparable to the heat shock treatment, which indicated that HSP72 expression was more stimulus specific. The protective effect of lactobacilli was further studied in IPEC-J2 under an enterotoxigenic Escherichia coli (ETEC) challenge. ETEC caused intestinal barrier destruction, as reflected by loss of cell–cell contact, reduced IPEC-J2 cell viability and transepithelial electrical resistance, and disruption of tight junction protein zonula occludens-1. In contrast, the L. reuteri treatment substantially counteracted these detrimental effects and preserved the barrier function. L. johnsonii and LGG also achieved barrier protection, partly by directly inhibiting ETEC attachment. Together, the results indicate that specific strains of Lactobacillus can enhance gut barrier function through cytoprotective HSP induction and fortify the cell protection against ETEC challenge through tight junction protein modulation and direct interaction with pathogens. PMID:25847917

  11. Characterization of a novel Lactobacillus species closely related to Lactobacillus johnsonii using a combination of molecular and comparative genomics methods

    PubMed Central

    2010-01-01

    Background Comparative genomic hybridization (CGH) constitutes a powerful tool for identification and characterization of bacterial strains. In this study we have applied this technique for the characterization of a number of Lactobacillus strains isolated from the intestinal content of rats fed with a diet supplemented with sorbitol. Results Phylogenetic analysis based on 16S rRNA gene, recA, pheS, pyrG and tuf sequences identified five bacterial strains isolated from the intestinal content of rats as belonging to the recently described Lactobacillus taiwanensis species. DNA-DNA hybridization experiments confirmed that these five strains are distinct but closely related to Lactobacillus johnsonii and Lactobacillus gasseri. A whole genome DNA microarray designed for the probiotic L. johnsonii strain NCC533 was used for CGH analysis of L. johnsonii ATCC 33200T, L. johnsonii BL261, L. gasseri ATCC 33323T and L. taiwanensis BL263. In these experiments, the fluorescence ratio distributions obtained with L. taiwanensis and L. gasseri showed characteristic inter-species profiles. The percentage of conserved L. johnsonii NCC533 genes was about 83% in the L. johnsonii strains comparisons and decreased to 51% and 47% for L. taiwanensis and L. gasseri, respectively. These results confirmed the separate status of L. taiwanensis from L. johnsonii at the level of species, and also that L. taiwanensis is closer to L. johnsonii than L. gasseri is to L. johnsonii. Conclusion Conventional taxonomic analyses and microarray-based CGH analysis have been used for the identification and characterization of the newly species L. taiwanensis. The microarray-based CGH technology has been shown as a remarkable tool for the identification and fine discrimination between phylogenetically close species, and additionally provided insight into the adaptation of the strain L. taiwanensis BL263 to its ecological niche. PMID:20849602

  12. Taxonomic and Strain-Specific Identification of the Probiotic Strain Lactobacillus rhamnosus 35 within the Lactobacillus casei Group?

    PubMed Central

    Coudeyras, Sophie; Marchandin, Hélène; Fajon, Céline; Forestier, Christiane

    2008-01-01

    Lactobacilli are lactic acid bacteria that are widespread in the environment, including the human diet and gastrointestinal tract. Some Lactobacillus strains are regarded as probiotics because they exhibit beneficial health effects on their host. In this study, the long-used probiotic strain Lactobacillus rhamnosus 35 was characterized at a molecular level and compared with seven reference strains from the Lactobacillus casei group. Analysis of rrn operon sequences confirmed that L. rhamnosus 35 indeed belongs to the L. rhamnosus species, and both temporal temperature gradient gel electrophoresis and ribotyping showed that it is closer to the probiotic strain L. rhamnosus ATCC 53103 (also known as L. rhamnosus GG) than to the species type strain. In addition, L. casei ATCC 334 gathered in a coherent cluster with L. paracasei type strains, unlike L. casei ATCC 393, which was closer to L. zeae; this is evidence of the lack of relatedness between the two L. casei strains. Further characterization of the eight strains by pulsed-field gel electrophoresis repetitive DNA element-based PCR identified distinct patterns for each strain, whereas two isolates of L. rhamnosus 35 sampled 40 years apart could not be distinguished. By subtractive hybridization using the L. rhamnosus GG genome as a driver, we were able to isolate five L. rhamnosus 35-specific sequences, including two phage-related ones. The primer pairs designed to amplify these five regions allowed us to develop rapid and highly specific PCR-based identification methods for the probiotic strain L. rhamnosus 35. PMID:18326671

  13. Expression of Clostridium thermocellum endoglucanase gene in Lactobacillus gasseri and Lactobacillus johnsonii and characterization of the genetically modified probiotic lactobacilli.

    PubMed

    Cho, J S; Choi, Y J; Chung, D K

    2000-04-01

    Endoglucanase A from Clostridium thermocellum resistant to pancreatic proteinase was selected out of a range of microbial cellulases expressed in lactobacilli. Two Lactobacillus-E. coli expression vectors, harboring the endoglucanase gene from C. thermocellum under the control of its own promoter (pSD1) and the Lactococcus lactis lac A promoter (pSD2), were constructed separately. Intestinal Lactobacillus strains, L. gasseri and L. johnsonii, were electrotransformed with pSD1 and pSD2, and the stability of each plasmid was evaluated. The endoglucanase activities of 0.722 and 0.759 U/ml were respectively found in culture medium of L. gasseri and L. johnsonii containing pSD1, and of 0.407 U/ml in medium of L. gasseri harboring pSD2. When the probiotic characteristics such as acid-tolerance, bile-salt tolerance, and antibiotic susceptibility were investigated, L. gasseri and L. johnsonii were resistant to low pHs of 2 and 3. Also, L. johnsonii was bile-salt resistant in the presence of 0.5% oxgall and porcine bile extract. L. johnsonii and L. gasseri showed a rather homogeneous resistant pattern against tested antibiotics. Both strains were resistant to amikacin, bacitracin, gentamicin, streptomycin, kanamycin, and colistin. PMID:10688695

  14. Comparative Genomic and Functional Analysis of Lactobacillus casei and Lactobacillus rhamnosus Strains Marketed as Probiotics

    PubMed Central

    Douillard, François P.; Ribbera, Angela; Järvinen, Hanna M.; Kant, Ravi; Pietilä, Taija E.; Randazzo, Cinzia; Paulin, Lars; Laine, Pia K.; Caggia, Cinzia; von Ossowski, Ingemar; Reunanen, Justus; Satokari, Reetta; Salminen, Seppo; Palva, Airi

    2013-01-01

    Four Lactobacillus strains were isolated from marketed probiotic products, including L. rhamnosus strains from Vifit (Friesland Campina) and Idoform (Ferrosan) and L. casei strains from Actimel (Danone) and Yakult (Yakult Honsa Co.). Their genomes and phenotypes were characterized and compared in detail with L. casei strain BL23 and L. rhamnosus strain GG. Phenotypic analysis of the new isolates indicated differences in carbohydrate utilization between L. casei and L. rhamnosus strains, which could be linked to their genotypes. The two isolated L. rhamnosus strains had genomes that were virtually identical to that of L. rhamnosus GG, testifying to their genomic stability and integrity in food products. The L. casei strains showed much greater genomic heterogeneity. Remarkably, all strains contained an intact spaCBA pilus gene cluster. However, only the L. rhamnosus strains produced mucus-binding SpaCBA pili under the conditions tested. Transcription initiation mapping demonstrated that the insertion of an iso-IS30 element upstream of the pilus gene cluster in L. rhamnosus strains but absent in L. casei strains had constituted a functional promoter driving pilus gene expression. All L. rhamnosus strains triggered an NF-?B response via Toll-like receptor 2 (TLR2) in a reporter cell line, whereas the L. casei strains did not or did so to a much lesser extent. This study demonstrates that the two L. rhamnosus strains isolated from probiotic products are virtually identical to L. rhamnosus GG and further highlights the differences between these and L. casei strains widely marketed as probiotics, in terms of genome content, mucus-binding and metabolic capacities, and host signaling capabilities. PMID:23315726

  15. A simple identification method for vaginal secretions using relative quantification of Lactobacillus DNA.

    PubMed

    Doi, Masanori; Gamo, Shinsuke; Okiura, Tatsuyuki; Nishimukai, Hiroaki; Asano, Migiwa

    2014-09-01

    In criminal investigations there are some cases in which identifying the presence of vaginal secretions provides crucial evidence in proving sexual assault. However, there are no methods for definitively identifying vaginal secretions. In the present study, we focused on Lactobacillus levels in vaginal secretions and developed a novel identification method for vaginal secretions by relative quantification based on real time PCR. We designed a Lactobacillus conserved region primer pair (LCP) by aligning 16S rRNA gene sequences from major vaginal Lactobacillus species (Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus iners and Lactobacillus jensenii), and selected the human specific primer pair (HSP) as an endogenous control for relative quantification. As a result, the ?Ct (?Ct=Ct[LCP]-Ct[HSP]) values of vaginal secretions (11 out of 12 samples) were significantly lower than those of saliva, semen and skin surface samples, and it was possible to discriminate between vaginal secretions and other body fluids. For the one remaining sample, it was confirmed that the predominant species in the microflora was not of the Lactobacillus genus. The ?Ct values in this study were calculated when the total DNA input used from the vaginal secretions was 10pg or more. Additionally, the ?Ct values of samples up to 6-months-old, which were kept at room temperature, remained unchanged. Thus, we concluded in this study that the simple ?Ct method by real time PCR is a useful tool for detecting the presence of vaginal secretions. PMID:24905338

  16. Growth of Lactobacillus paracasei ATCC334 in a cheese model system: A biochemical approach

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Growth of Lactobacillus paracasei ATCC 334, in a cheese-ripening model system based upon a medium prepared from ripening Cheddar cheese extract (CCE) was evaluated. Lactobacillus paracasei ATCC 334 grows in CCE made from cheese ripened for 2 (2mCCE), 6 (6mCCE), and 8 (8mCCE) mo, to final cell densit...

  17. Draft Genome Sequence of Lactobacillus gorillae Strain KZ01T, Isolated from a Western Lowland Gorilla

    PubMed Central

    Tsuchida, Sayaka; Nezuo, Maiko; Tsukahara, Masatoshi; Ogura, Yoshitoshi; Hayashi, Tetsuya

    2015-01-01

    Here, we report the draft genome sequence of Lactobacillus gorillae strain KZ01T isolated from a western lowland gorilla (Gorilla gorilla gorilla). This genome sequence will be helpful for the comparative genomics between human and nonhuman primate-associated Lactobacillus. PMID:26472838

  18. Draft Genome Sequence of Lactobacillus gorillae Strain KZ01T, Isolated from a Western Lowland Gorilla.

    PubMed

    Tsuchida, Sayaka; Nezuo, Maiko; Tsukahara, Masatoshi; Ogura, Yoshitoshi; Hayashi, Tetsuya; Ushida, Kazunari

    2015-01-01

    Here, we report the draft genome sequence of Lactobacillus gorillae strain KZ01(T) isolated from a western lowland gorilla (Gorilla gorilla gorilla). This genome sequence will be helpful for the comparative genomics between human and nonhuman primate-associated Lactobacillus. PMID:26472838

  19. Evaluation of Antibacterial Activity of Lactobacillus Spp. on Selected Food Spoilage Bacteria.

    PubMed

    Sharma, Anurag; Gupta, Piyush; Bhattacharya, Susinjan

    2015-01-01

    This study was done to isolate Lactobacillus species from curd, amla/Indian gooseberry and orange and to assess their antagonistic ability against selected food spoilage bacteria, Escherichia coli, Pseudomonas spp. and Bacillus spp. isolated from natural food sources. In the approaches used, native Lactobacillus spp. were isolated from amla, orange and curd and identified by standard microbiological methods. Their antagonistic affect was tested by disc diffusion tests against three selected test isolates, Escherichia coli, Pseudomonas and Bacillus spp. isolated from tomato, pumpkin, cauliflower, lady's finger, carrot, and milk. There are recent patents also suggesting use of novel strains of Lactobacillus for microbial antagonism. In our present work, the lactobacilli isolated from different food sources showed varied ability to inhibit the growth of test isolates. The growth of test isolates was inhibited by Lactobacillus isolates with one of the Lactobacillus isolate from amla being the most potent inhibitor. PMID:25751004

  20. Lactobacillus species as biomarkers and agents that can promote various aspects of vaginal health

    PubMed Central

    Petrova, Mariya I.; Lievens, Elke; Malik, Shweta; Imholz, Nicole; Lebeer, Sarah

    2015-01-01

    The human body is colonized by a vast number of microorganisms collectively referred to as the human microbiota. One of the main microbiota body sites is the female genital tract, commonly dominated by Lactobacillus spp., in approximately 70% of women. Each individual species can constitute approximately 99% of the ribotypes observed in any individual woman. The most frequently isolated species are Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus jensenii and Lactobacillus iners. Residing at the port of entry of bacterial and viral pathogens, the vaginal Lactobacillus species can create a barrier against pathogen invasion since mainly products of their metabolism secreted in the cervicovaginal fluid can play an important role in the inhibition of bacterial and viral infections. Therefore, a Lactobacillus-dominated microbiota appears to be a good biomarker for a healthy vaginal ecosystem. This balance can be rapidly altered during processes such as menstruation, sexual activity, pregnancy and various infections. An abnormal vaginal microbiota is characterized by an increased diversity of microbial species, leading to a condition known as bacterial vaginosis. Information on the vaginal microbiota can be gathered from the analysis of cervicovaginal fluid, by using the Nugent scoring or the Amsel's criteria, or at the molecular level by investigating the number and type of Lactobacillus species. However, when translating this to the clinical setting, it should be noted that the absence of a Lactobacillus-dominated microbiota does not appear to directly imply a diseased condition or dysbiosis. Nevertheless, the widely documented beneficial role of vaginal Lactobacillus species demonstrates the potential of data on the composition and activity of lactobacilli as biomarkers for vaginal health. The substantiation and further validation of such biomarkers will allow the design of better targeted probiotic strategies. PMID:25859220

  1. Draft Genome Sequences of Lactobacillus equicursoris CIP 110162T and Lactobacillus sp. Strain CRBIP 24.137, Isolated from Thoroughbred Racehorse Feces and Human Urine, Respectively

    PubMed Central

    Cousin, Sylvie; Loux, Valentin; Ma, Laurence; Creno, Sophie; Clermont, Dominique; Bizet, Chantal

    2013-01-01

    We report the draft genome sequences of strain Lactobacillus equicursoris CIP 110162T, isolated from racehorse breed feces, and Lactobacillus sp. strain CRBIP 24.137, isolated from human urine; the two strains are closely related. The total lengths of the 116 and 62 scaffolds are about 2.157 and 2.358 Mb, with G+C contents of 46 and 45% and 2,279 and 2,342 coding sequences (CDSs), respectively. PMID:23969063

  2. Nanosurgery: Observation of Peptidoglycan Strands in Lactobacillus helveticus Cell Walls

    E-print Network

    Max Firtel; Grant Henderson; Igor Sokolov

    2004-07-05

    The internal cell wall structure of the bacterium Lactobacillus helveticus has been observed in situ in aqueous solution using an atomic force microscope (AFM). The AFM tip was used not only for imaging but presumably to remove mechanically large patches of the outer cell wall after appropriate chemical treatment, which typically leaves the bacteria alive. The surface exposed after such a surgery revealed ca. 26 nm thick twisted strands within the cell wall. The structure and location of the observed strands are consistent with the glycan backbone of peptidoglycan fibers that give strength to the cell wall. The found structural organization of these fibers has not been observed previously.

  3. Endocarditis of the native aortic valve caused by Lactobacillus jensenii.

    PubMed

    Patnaik, Soumya; Davila, Carlos Daniel; Chennupati, Anupama; Rubin, Alexander

    2015-01-01

    Lactobacilli are Gram-positive anaerobic rods or coccobacilli, commonly found as commensals in human mucosa. Rarely, they can cause serious infections such as infective endocarditis (IE), and the most frequently implicated species causing serious infections are L. casei and L. rhamnosus. IE caused by Lactobacillus jensenii is very rare, with only six reported cases so far, to the best of our knowledge. We present a case of native aortic valve endocarditis caused by L. jensenii, complicated by root abscess and complete heart block, and requiring emergent surgical intervention. PMID:25750218

  4. Radiation resistance of lactobacilli isolated from radurized meat relative to growth and environment. [Lactobacillus sake; Lactobacillus curvatus; Lactobacillus farciminis; Staphylococcus aureus; Salmonella typimurium

    SciTech Connect

    Hastings, J.W.; Holzapfel, W.H.; Niemand, J.G.

    1986-10-01

    Of 113 lactobacilli isolated from radurized (5 kGy) minced meat, 7 Lactobacillus sake strains, 1 L. curvatus strain, and 1 L. farciminis strain were used for radiation resistance studies in a semisynthetic substrate (i.e., modified MRS broth). Five reference Lactobacillus spp. one Staphylococcus aureus strain, and one Salmonella typhimurium strain were used for comparative purposes. All L. sake isolates exhibited the phenomenon of being more resistant to gamma-irradiation in the exponential (log) phase than in the stationary phase of their growth cycles by a factor of 28%. Four reference strains also exhibited this phenomenon, with L. sake (DSM 20017) showing a 68% increase in resistance in the log phase over the stationary phase. This phenomenon was not common to all bacteria tested and is not common to all strains with high radiation resistance. Four L. sake isolates and three reference strains were used in radiation sensitivity testing in a natural food system (i.e., meat). The bacteria were irradiated in minced meat and packaged under four different conditions (air, vacuum, CO/sub 2/, and N/sub 2/). Organisms exhibited the highest death rate (lowest D/sub 10/ values (doses required to reduce the logarithm of the bacterial population by 1) under CO/sub 2/ packaging conditions, but resistance to irradiation was increased under N/sub 2/. The D/sup 10/ values of the isolates were generally greater than those of the reference strains. The D/sup 10/ values were also higher (approximately two times) in meat than in a semisynthetic growth medium.

  5. The predominance of Lactobacillus sanfranciscensis in French organic sourdoughs and its impact on related bread characteristics.

    PubMed

    Lhomme, Emilie; Orain, Servane; Courcoux, Philippe; Onno, Bernard; Dousset, Xavier

    2015-11-20

    Fourteen bakeries located in different regions of France were selected. These bakers use natural sourdough and organic ingredients. Consequently, different organic sourdoughs used for the manufacture of French bread were studied by the enumeration of lactic acid bacteria (LAB) and 16S rRNA sequencing of the isolates. In addition, after DNA extraction the bacterial diversity was assessed by pyrosequencing of the 16S rDNA V1-V3 region. Although LAB counts showed significant variations (7.6-9.5log10CFU/g) depending on the sourdough studied, their identification through a polyphasic approach revealed a large predominance of Lactobacillus sanfranciscensis in all samples. In ten sourdoughs, both culture and independent methods identified L. sanfranciscensis as the dominant LAB species identified. In the remaining sourdoughs, culture methods identified 30-80% of the LAB as L. sanfranciscensis whereas more than 95% of the reads obtained by pyrosequencing belonged to L. sanfranciscensis. Other sub-dominant species, such as Lactobacillus curvatus, Lactobacillus hammesii, Lactobacillus paralimentarius, Lactobacillus plantarum, Lactobacillus pentosus, and Lactobacillus sakei, were also identified. Quantification of L. sanfranciscensis by real-time PCR confirmed the predominance of this species ranging from 8.24 to 10.38log10CFU/g. Regarding the acidification characteristics, sourdough and related bread physico-chemical characteristics varied, questioning the involvement of sub-dominant species or L. sanfranciscensis intra-species diversity and/or the role of the baker's practices. PMID:26051957

  6. B cells are not essential for Lactobacillus-mediated protection against lethal pneumovirus infection.

    PubMed

    Percopo, Caroline M; Dyer, Kimberly D; Garcia-Crespo, Katia E; Gabryszewski, Stanislaw J; Shaffer, Arthur L; Domachowske, Joseph B; Rosenberg, Helene F

    2014-06-01

    We have shown previously that priming of respiratory mucosa with live Lactobacillus species promotes robust and prolonged survival from an otherwise lethal infection with pneumonia virus of mice, a property known as heterologous immunity. Lactobacillus priming results in a moderate reduction in virus recovery and a dramatic reduction in virus-induced proinflammatory cytokine production; the precise mechanisms underlying these findings remain to be elucidated. Because B cells have been shown to promote heterologous immunity against respiratory virus pathogens under similar conditions, in this study we explore the role of B cells in Lactobacillus-mediated protection against acute pneumovirus infection. We found that Lactobacillus-primed mice feature elevated levels of airway Igs IgG, IgA, and IgM and lung tissues with dense, B cell (B220(+))-enriched peribronchial and perivascular infiltrates with germinal centers consistent with descriptions of BALT. No B cells were detected in lung tissue of Lactobacillus-primed B cell deficient ?MT mice or Jh mice, and Lactobacillus-primed ?MT mice had no characteristic infiltrates or airway Igs. Nonetheless, we observed diminished virus recovery and profound suppression of virus-induced proinflammatory cytokines CCL2, IFN-?, and CXCL10 in both wild-type and Lactobacillus-primed ?MT mice. Furthermore, Lactobacillus plantarum-primed, B cell-deficient ?MT and Jh mice were fully protected from an otherwise lethal pneumonia virus of mice infection, as were their respective wild-types. We conclude that B cells are dispensable for Lactobacillus-mediated heterologous immunity and were not crucial for promoting survival in response to an otherwise lethal pneumovirus infection. PMID:24748495

  7. Debittering of Protein Hydrolysates by Lactobacillus LBL-4 Aminopeptidase

    PubMed Central

    Tchorbanov, Bozhidar; Marinova, Margarita; Grozeva, Lydia

    2011-01-01

    Yoghurt strain Lactobacillus LBL-4 cultivated for 8–10?h at pH ~6.0 was investigated as a considerable food-grade source of intracellular aminopeptidase. Cell-free extract manifesting >200?AP U/l was obtained from cells harvested from 1?L culture media. Subtilisin-induced hydrolysates of casein, soybean isolate, and Scenedesmus cell protein with degree of hydrolysis 20–22% incubated at 45°C for 10?h by 10 AP?U/g peptides caused an enlarging of DH up to 40–42%, 46–48%, and 38–40% respectively. The DH increased rapidly during the first 4?h, but gel chromatography studies on BioGel P-2 showed significant changes occurred during 4–10?h of enzyme action when the DH increased gradually. After the digestion, the remained AP activity can be recovered by ultrafiltration (yield 40–50%). Scenedesmus protein hydrolysate with DH 20% was inoculated by Lactobacillus LBL-4 cells, and after 72?h cultivation the DH reached 32%. The protein hydrolysates (DH above 40%) obtained from casein and soybean isolate (high Q value) demonstrated a negligible bitterness while Scenedesmus protein hydrolysates (low Q value) after both treatments were free of bitterness. PMID:21876793

  8. Bioactivity characterization of Lactobacillus strains isolated from dairy products.

    PubMed

    Haghshenas, Babak; Nami, Yousef; Haghshenas, Minoo; Abdullah, Norhafizah; Rosli, Rozita; Radiah, Dayang; Yari Khosroushahi, Ahmad

    2015-10-01

    This study aimed to find candidate strains of Lactobacillus isolated from sheep dairy products (yogurt and ewe colostrum) with probiotic and anticancer activity. A total of 100 samples were randomly collected from yogurt and colostrum and 125 lactic acid bacteria were isolated. Of these, 17 Lactobacillus strains belonging to five species (L. delbrueckii, L. plantarum, L. rhamnosus, L. paracasei, and L. casei) were identified. L. plantarum 17C and 13C, which isolated from colostrums, demonstrated remarkable results such as resistant to low pH and high concentrations of bile salts, susceptible to some antibiotics and good antimicrobial activity that candidate them as potential probiotics. Seven strains (1C, 5C, 12C, 13C, 17C, 7M, and 40M), the most resistant to simulated digestion, were further investigated to evaluate their capability to adhere to human intestinal Caco-2 cells. L. plantarum 17C was the most adherent strain. The bioactivity assessment of L. plantarum 17C showed anticancer effects via the induction of apoptosis on HT-29 human cancer cells and negligible side effects on one human epithelial normal cell line (FHs 74). The metabolites produced by this strain can be used as alternative pharmaceutical compounds with promising therapeutic indices because they are not cytotoxic to normal mammalian cells. PMID:26219634

  9. Bioactivity characterization of Lactobacillus strains isolated from dairy products

    PubMed Central

    Haghshenas, Babak; Nami, Yousef; Haghshenas, Minoo; Abdullah, Norhafizah; Rosli, Rozita; Radiah, Dayang; Yari Khosroushahi, Ahmad

    2015-01-01

    This study aimed to find candidate strains of Lactobacillus isolated from sheep dairy products (yogurt and ewe colostrum) with probiotic and anticancer activity. A total of 100 samples were randomly collected from yogurt and colostrum and 125 lactic acid bacteria were isolated. Of these, 17 Lactobacillus strains belonging to five species (L. delbrueckii, L. plantarum, L. rhamnosus, L. paracasei, and L. casei) were identified. L. plantarum 17C and 13C, which isolated from colostrums, demonstrated remarkable results such as resistant to low pH and high concentrations of bile salts, susceptible to some antibiotics and good antimicrobial activity that candidate them as potential probiotics. Seven strains (1C, 5C, 12C, 13C, 17C, 7M, and 40M), the most resistant to simulated digestion, were further investigated to evaluate their capability to adhere to human intestinal Caco-2 cells. L. plantarum 17C was the most adherent strain. The bioactivity assessment of L. plantarum 17C showed anticancer effects via the induction of apoptosis on HT-29 human cancer cells and negligible side effects on one human epithelial normal cell line (FHs 74). The metabolites produced by this strain can be used as alternative pharmaceutical compounds with promising therapeutic indices because they are not cytotoxic to normal mammalian cells. PMID:26219634

  10. Lactobacillus psittaci sp. nov., isolated from a hyacinth macaw (Anodorhynchus hyacinthinus).

    PubMed

    Lawson, P A; Wacher, C; Hansson, I; Falsen, E; Collins, M D

    2001-05-01

    A Gram-positive, non-spore-forming, catalase-negative, facultatively anaerobic coccibacillus to rod-shaped bacterium isolated from a parrot was characterized using phenotypic and molecular taxonomic methods. The unknown bacterium phenotypically resembled lactobacilli and comparative 16S rRNA gene sequencing demonstrated that the organism represents a distinct subline within the Lactobacillus delbrueckii rRNA cluster of the genus. 16S rRNA sequence divergence values of > 6% with recognized Lactobacillus species clearly demonstrated the phylogenetic separateness of the parrot bacterium. On the basis of phylogenetic evidence and the phenotypic distinctiveness of the unknown bacterium, a new species, Lactobacillus psittaci sp. nov., is proposed. The type strain of Lactobacillus psittaci is CCUG 42378T (= CIP 106492T). PMID:11411722

  11. Efficacy of supercritical carbon dioxide for inactivating Lactobacillus plantarum in apple cider

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Juice makers have traditionally used thermal pasteurization to prevent deterioration by spoilage bacteria such as Lactobacillus plantarum; however this thermal processing causes adverse effects on product quality such as undesirable taste and destruction of heat sensitive nutrients. For this reason,...

  12. Bacteriophage endolysins expressed in yeast kill strains of Lactobacillus that contaminate fermentations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    One of the challenges facing the fuel ethanol industry is the management of bacterial contamination during fermentation. Species of Lactobacillus are the predominant contaminants that reduce ethanol yields and cause “stuck” fermentations, decreasing the profitability of biofuel production with expen...

  13. Inactivation of Lactobacillus leichmannii ribonucleotide reductase by F2CTP: covalent modification

    E-print Network

    Lohman, Gregory J. S.

    Ribonucleotide reductase (RNR) from Lactobacillus leichmannii, a 76 kDa monomer using adenosylcobalamin (AdoCbl) as a cofactor, catalyzes the conversion of nucleoside triphosphates to deoxynucleotides and is rapidly (<30 ...

  14. B cells are not essential for Lactobacillus-mediated protection against lethal pneumovirus infection*

    PubMed Central

    Percopo, Caroline M.; Dyer, Kimberly D.; Garcia-Crespo, Katia E.; Gabryszewski, Stanislaw J.; Shaffer, Arthur L.; Domachowske, Joseph B.; Rosenberg, Helene F.

    2014-01-01

    We have shown previously that priming of respiratory mucosa with live Lactobacillus species promotes robust and prolonged survival from an otherwise lethal infection with pneumonia virus of mice (PVM), a property known as heterologous immunity. Lactobacillus-priming results in a moderate reduction in virus recovery and a dramatic reduction in virus-induced proinflammatory cytokine production; the precise mechanisms underlying these findings remain to be elucidated. As B cells have been shown to promote heterologous immunity against respiratory virus pathogens under similar conditions, here we explore the role of B cells in Lactobacillus-mediated protection against acute pneumovirus infection. We found that Lactobacillus-primed mice feature elevated levels of airway immunoglobulins IgG, IgA and IgM and lung tissues with dense, B cell (B220+) enriched peribronchial and perivascular infiltrates with germinal centers consistent with descriptions of bronchus-associated lymphoid tissue. No B cells were detected in lung tissue of Lactobacillus-primed B-cell deficient ?MT mice or Jh mice, and Lactobacillus-primed ?MT mice had no characteristic infiltrates or airway immunoglobulins. Nonetheless, we observed diminished virus recovery and profound suppression of virus-induced proinflammatory cytokines CCL2, IFN-gamma, and CXCL10 in both wild-type and Lactobacillus-primed ?MT mice. Furthermore, L. plantarum-primed, B-cell deficient ?MT and Jh mice were fully protected from an otherwise lethal PVM infection, as were their respective wild-types. We conclude that B cells are dispensable for Lactobacillus-mediated heterologous immunity and were not crucial for promoting survival in response to an otherwise lethal pneumovirus infection. PMID:24748495

  15. Evaluation of profertility effect of probiotic Lactobacillus plantarum 2621 in a murine model

    PubMed Central

    Bhandari, Praveen; Prabha, Vijay

    2015-01-01

    Background & objectives: Urogenital infections of bacterial origin have a high incidence among the female population at reproductive age, affecting the fertility. Strains of Escherichia coli can colonize the vagina and replace natural microflora. Lactobacillus the predominant vaginal microorganism in healthy women, maintains the acidic vaginal pH which inhibits pathogenic microorganisms. Studies on Lactobacillus have shown that these can inhibit E. coli growth and vaginal colonization. An alternative therapeutic approach to antimicrobial therapy is to re-establish Lactobacillus in this microbiome through probiotic administration to resurge fertility. Therefore, the aim of the present study was to determine the capability of L. plantarum 2621 strain with probiotic properties, to prevent the vaginal colonization of E. coli causing agglutination of sperms and to evaluate its profertility effect in a murine model. Methods: Screened mice were divided into five groups i.e. control group, E. coli group, Lactobacillus group, prophylactic and therapeutic groups. The control group was infused with 20 µl PBS, E.coli group was administered with 106 cfu/20 µl E. coli, and probiotic group was administered with Lactobacillus (108 cfu/20 µl) for 10 consecutive days. In prophylactic group, the vagina was colonized with 10 consecutive doses of Lactobacillus (108 cfu/20 µl). After 24 h, it was followed by 10 day intravaginal infection with E. coli (106 cfu/20 µl) whereas for the therapeutic group vagina was colonized with (106 cfu/20 µl) E. coli for 10 consecutive days, followed by 10 day intravaginal administration with Lactobacillus after 24 h. Results: Upon mating and completion of gestation period, control, probiotic and the therapeutic groups had litters in contrast to the prophylactic group and the group administered with E. coli. Interpretation & conclusions: Results indicated that Lactobacillus intermitted colonization of pathogenic strains that resulted in reinforcement of natural microflora and resurge fertility. PMID:26261170

  16. Lactobacillus-mediated priming of the respiratory mucosa protects against lethal pneumovirus infection.

    PubMed

    Gabryszewski, Stanislaw J; Bachar, Ofir; Dyer, Kimberly D; Percopo, Caroline M; Killoran, Kristin E; Domachowske, Joseph B; Rosenberg, Helene F

    2011-01-15

    The inflammatory response to respiratory virus infection can be complex and refractory to standard therapy. Lactobacilli, when targeted to the respiratory epithelium, are highly effective at suppressing virus-induced inflammation and protecting against lethal disease. Specifically, wild-type mice primed via intranasal inoculation with live or heat-inactivated Lactobacillus plantarum or Lactobacillus reuteri were completely protected against lethal infection with the virulent rodent pathogen, pneumonia virus of mice; significant protection (60% survival) persisted for at least 13 wk. Protection was not unique to Lactobacillus species, and it was also observed in response to priming with nonpathogenic Gram-positive Listeria innocua. Priming with live lactobacilli resulted in diminished granulocyte recruitment, diminished expression of multiple proinflammatory cytokines (CXCL10, CXCL1, CCL2, and TNF), and reduced virus recovery, although we have demonstrated clearly that absolute virus titer does not predict clinical outcome. Lactobacillus priming also resulted in prolonged survival and protection against the lethal sequelae of pneumonia virus of mice infection in MyD88 gene-deleted (MyD88(-/-)) mice, suggesting that the protective mechanisms may be TLR-independent. Most intriguing, virus recovery and cytokine expression patterns in Lactobacillus-primed MyD88(-/-) mice were indistinguishable from those observed in control-primed MyD88(-/-) counterparts. In summary, we have identified and characterized an effective Lactobacillus-mediated innate immune shield, which may ultimately serve as critical and long-term protection against infection in the absence of specific antiviral vaccines. PMID:21169550

  17. Anti-Inflammatory Activity of Lactobacillus on Carrageenan-Induced Paw Edema in Male Wistar Rats

    PubMed Central

    Amdekar, Sarika; Roy, Purabi; Singh, Vinod; Kumar, Avnish; Singh, Rambir; Sharma, Poonam

    2012-01-01

    Introduction. Lactobacillus casei and Lactobacillus acidophilus were used to assess the anti-inflammatory properties in carrageenan induced acute inflammatory model. Materials and Methods. Diclofenac sodium was used as standard drug at concentration of 150?mg/kg of body weight. Culture of Lactobacillus??2 × 107?CFU/ml was given orally. Edema was induced with 1% carrageenan to all the groups after one hour of the oral treatments. Paw thickness was checked at t = 1, 2, 3, 4, 5, and 24 hours. Stair climbing score and motility score were assessed at t = 24 hours. Cytokines assay for IL-6, IL-10, and TNF-? was performed on serum samples. Results. Lactobacillus showed a statistically significant decrease in paw thickness at P < 0.001. L. acidophilus and L. casei decreased by 32% and 28% in paw thickness. They both significantly increased the stair climbing and motility score. Lactobacillus treatment significantly downregulated IL-6 and TNF-? while upregulated IL-10 at P < 0.0001. Conclusion. L. casei and L. acidophilus significantly decreased the inflammatory reactions induced by carrageenan. This study has also proposed that Lactobacillus ameliorated the inflammatory reaction by downregulating the proinflammatory cytokines pathway. PMID:22518342

  18. Lactobacillus reuteri 100-23 Modulates Urea Hydrolysis in the Murine Stomach

    PubMed Central

    Wilson, Charlotte M.; Loach, Diane; Lawley, Blair; Bell, Tracey; Sims, Ian M.; O'Toole, Paul W.; Zomer, Aldert

    2014-01-01

    Comparisons of in vivo (mouse stomach) and in vitro (laboratory culture) transcriptomes of Lactobacillus reuteri strain 100-23 were made by microarray analysis. These comparisons revealed the upregulation of genes associated with acid tolerance, including urease production, in the mouse stomach. Inactivation of the ureC gene reduced the acid tolerance of strain 100-23 in vitro, and the mutant was outcompeted by the wild type in the gut of ex-Lactobacillus-free mice. Urine analysis showed that stable isotope-labeled urea, administered by gavage, was metabolized to a greater extent in Lactobacillus-free mice than animals colonized by strain 100-23. This surprising observation was associated with higher levels of urease activity and fecal-type bacteria in the stomach digesta of Lactobacillus-free mice. Despite the modulation of urea hydrolysis in the stomach, recycling of urea nitrogen in the murine host was not affected since the essential amino acid isoleucine, labeled with a stable isotope, was detected in the livers of both Lactobacillus-free and 100-23-colonized animals. Therefore, our experiments reveal a new and unexpected impact of Lactobacillus colonization on urea hydrolysis in the murine gut. PMID:25063664

  19. Lactobacillus sicerae sp. nov., a lactic acid bacterium isolated from Spanish natural cider.

    PubMed

    Puertas, Ana Isabel; Arahal, David R; Ibarburu, Idoia; Elizaquível, Patricia; Aznar, Rosa; Dueñas, M Teresa

    2014-09-01

    Strains CUPV261(T) and CUPV262 were isolated from ropy natural ciders of the Basque Country, Spain, in 2007. Cells are Gram-stain positive, non-spore-forming, motile rods, facultative anaerobes and catalase-negative. The strains are obligately homofermentative (final product dl-lactate) and produce exopolysaccharides from sucrose. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the highest similarity to both isolates corresponded to the type strain of Lactobacillus vini (99.1?%), followed by Lactobacillus satsumensis (96.4?%), and Lactobacillus oeni (96.2?%), and for all other established species, 16S rRNA gene sequence similarities were below 96?%. The species delineation of strains CUPV261(T) and CUPV262 was evaluated through RAPD fingerprinting. In addition, a random partial genome pyrosequencing approach was performed on strain CUPV261(T) in order to compare it with the genome sequence of Lactobacillus vini DSM 20605(T) and calculate indexes of average nucleotide identity (ANI) between them. Results permit the conclusion that strains CUPV261(T) and CUPV262 represent a novel species of the genus Lactobacillus, for which the name Lactobacillus sicerae sp. nov. is proposed. The type strain is CUPV261(T) (?=?CECT 8227(T)?=?KCTC 21012(T)). PMID:24899655

  20. The behaviour of whey protein isolate in protecting Lactobacillus plantarum.

    PubMed

    Khem, Sarim; Small, Darryl M; May, Bee K

    2016-01-01

    There is increasing evidence that whey protein isolates (WPI), can be utilised to encapsulate and protect bioactive substances, including lactic acid bacteria, due to their physicochemical properties. However, little is known about what happens in the immediate vicinity of the cells. This study examined the protective behaviour of WPI for two strains of Lactobacillus plantarum, A17 and B21, during spray drying. B21 was found to be more hydrophobic than A17 and required 50% of the amount of WPI to provide comparably high survival (? 90%). We hypothesise that WPI protects the hydrophobic bacteria by initial attachment to the unfolded whey protein due to hydrophobic interactions followed by adhesion to the proteins, resulting in cells being embedded within the walls of the capsules. The encapsulated strains had a moisture content of approximately 5.5% and during storage trials at 20 °C retained viability for at least eight weeks. PMID:26213030

  1. Characterization of a Feruloyl Esterase from Lactobacillus plantarum

    PubMed Central

    Esteban-Torres, María; Reverón, Inés; Mancheño, José Miguel; de las Rivas, Blanca

    2013-01-01

    Lactobacillus plantarum is frequently found in the fermentation of plant-derived food products, where hydroxycinnamoyl esters are abundant. L. plantarum WCFS1 cultures were unable to hydrolyze hydroxycinnamoyl esters; however, cell extracts from the strain partially hydrolyze methyl ferulate and methyl p-coumarate. In order to discover whether the protein Lp_0796 is the enzyme responsible for this hydrolytic activity, it was recombinantly overproduced and enzymatically characterized. Lp_0796 is an esterase that, among other substrates, is able to efficiently hydrolyze the four model substrates for feruloyl esterases (methyl ferulate, methyl caffeate, methyl p-coumarate, and methyl sinapinate). A screening test for the detection of the gene encoding feruloyl esterase Lp_0796 revealed that it is generally present among L. plantarum strains. The present study constitutes the description of feruloyl esterase activity in L. plantarum and provides new insights into the metabolism of hydroxycinnamic compounds in this bacterial species. PMID:23793626

  2. Genetic Determinants of Reutericyclin Biosynthesis in Lactobacillus reuteri

    PubMed Central

    Lin, Xiaoxi B.; Lohans, Christopher T.; Duar, Rebbeca; Zheng, Jinshui; Vederas, John C.; Walter, Jens

    2015-01-01

    Reutericyclin is a unique antimicrobial tetramic acid produced by some strains of Lactobacillus reuteri. This study aimed to identify the genetic determinants of reutericyclin biosynthesis. Comparisons of the genomes of reutericyclin-producing L. reuteri strains with those of non-reutericyclin-producing strains identified a genomic island of 14 open reading frames (ORFs) including genes coding for a nonribosomal peptide synthetase (NRPS), a polyketide synthase (PKS), homologues of PhlA, PhlB, and PhlC, and putative transport and regulatory proteins. The protein encoded by rtcN is composed of a condensation domain, an adenylation domain likely specific for d-leucine, and a thiolation domain. rtcK codes for a PKS that is composed of a ketosynthase domain, an acyl-carrier protein domain, and a thioesterase domain. The products of rtcA, rtcB, and rtcC are homologous to the diacetylphloroglucinol-biosynthetic proteins PhlABC and may acetylate the tetramic acid moiety produced by RtcN and RtcK, forming reutericyclin. Deletion of rtcN or rtcABC in L. reuteri TMW1.656 abrogated reutericyclin production but did not affect resistance to reutericyclin. Genes coding for transport and regulatory proteins could be deleted only in the reutericyclin-negative L. reuteri strain TMW1.656?rtcN, and these deletions eliminated reutericyclin resistance. The genomic analyses suggest that the reutericyclin genomic island was horizontally acquired from an unknown source during a unique event. The combination of PhlABC homologues with both an NRPS and a PKS has also been identified in the lactic acid bacteria Streptococcus mutans and Lactobacillus plantarum, suggesting that the genes in these organisms and those in L. reuteri share an evolutionary origin. PMID:25576609

  3. Manufacture of probiotic Minas Frescal cheese with Lactobacillus casei Zhang.

    PubMed

    Dantas, Aline B; Jesus, Vitor F; Silva, Ramon; Almada, Carine N; Esmerino, E A; Cappato, Leandro P; Silva, Marcia C; Raices, Renata S L; Cavalcanti, Rodrigo N; Carvalho, Celio C; Sant'Ana, Anderson S; Bolini, Helena M A; Freitas, Monica Q; Cruz, Adriano G

    2016-01-01

    In this study, the addition of Lactobacillus casei Zhang in the manufacture of Minas Frescal cheese was investigated. Minas Frescal cheeses supplemented with probiotic bacteria (Lactobacillus casei Zhang) were produced by enzymatic coagulation and direct acidification and were subjected to physicochemical (pH, proteolysis, lactic acid, and acetic acid), microbiological (probiotic and lactic bacteria counts), and rheological analyses (uniaxial compression and creep test), instrumental color determination (luminosity, yellow intensity, and red intensity) and sensory acceptance test. The addition of L. casei Zhang resulted in low pH values and high proteolysis indexes during storage (from 5.38 to 4.94 and 0.470 to 0.702, respectively). Additionally, the cheese protocol was not a hurdle for growth of L. casei Zhang, as the population reached 8.16 and 9.02 log cfu/g by means of the direct acidification and enzymatic coagulation protocol, respectively, after 21d of refrigerated storage. The rheology data showed that all samples presented a more viscous-like behavior, which rigidity tended to decrease during storage and lower luminosity values were also observed. Increased consumer acceptance was observed for the control sample produced by direct acidification (7.8), whereas the cheeses containing L. casei Zhang presented lower values for all sensory attributes, especially flavor and overall liking (5.37 and 4.61 for enzymatic coagulation and 5.57 and 4.72 for direct acidification, respectively). Overall, the addition of L. casei Zhang led to changes in all parameters and affected negatively the sensory acceptance. The optimization of L. casei Zhang dosage during the manufacturing of probiotic Minas Frescal cheese should be performed. PMID:26519974

  4. Characterization of Two Virulent Phages of Lactobacillus plantarum

    PubMed Central

    Briggiler Marcó, Mariángeles; Garneau, Josiane E.; Tremblay, Denise; Quiberoni, Andrea

    2012-01-01

    We characterized two Lactobacillus plantarum virulent siphophages, ATCC 8014-B1 (B1) and ATCC 8014-B2 (B2), previously isolated from corn silage and anaerobic sewage sludge, respectively. Phage B2 infected two of the eight L. plantarum strains tested, while phage B1 infected three. Phage adsorption was highly variable depending on the strain used. Phage defense systems were found in at least two L. plantarum strains, LMG9211 and WCSF1. The linear double-stranded DNA genome of the pac-type phage B1 had 38,002 bp, a G+C content of 47.6%, and 60 open reading frames (ORFs). Surprisingly, the phage B1 genome has 97% identity with that of Pediococcus damnosus phage clP1 and 77% identity with that of L. plantarum phage JL-1; these phages were isolated from sewage and cucumber fermentation, respectively. The double-stranded DNA (dsDNA) genome of the cos-type phage B2 had 80,618 bp, a G+C content of 36.9%, and 127 ORFs with similarities to those of Bacillus and Lactobacillus strains as well as phages. Some phage B2 genes were similar to ORFs from L. plantarum phage LP65 of the Myoviridae family. Additionally, 6 tRNAs were found in the phage B2 genome. Protein analysis revealed 13 (phage B1) and 9 (phage B2) structural proteins. To our knowledge, this is the first report describing such high identity between phage genomes infecting different genera of lactic acid bacteria. PMID:23042172

  5. Lactic acid production from biomass-derived sugars via co-fermentation of Lactobacillus brevis and Lactobacillus plantarum.

    PubMed

    Zhang, Yixing; Vadlani, Praveen V

    2015-06-01

    Lignocellulosic biomass is an attractive alternative resource for producing chemicals and fuels. Xylose is the dominating sugar after hydrolysis of hemicellulose in the biomass, but most microorganisms either cannot ferment xylose or have a hierarchical sugar utilization pattern in which glucose is consumed first. To overcome this barrier, Lactobacillus brevis ATCC 367 was selected to produce lactic acid. This strain possesses a relaxed carbon catabolite repression mechanism that can use glucose and xylose simultaneously; however, lactic acid yield was only 0.52 g g(-1) from a mixture of glucose and xylose, and 5.1 g L(-1) of acetic acid and 8.3 g L(-1) of ethanol were also formed during production of lactic acid. The yield was significantly increased and ethanol production was significantly reduced if L. brevis was co-cultivated with Lactobacillus plantarum ATCC 21028. L. plantarum outcompeted L. brevis in glucose consumption, meaning that L. brevis was focused on converting xylose to lactic acid and the by-product, ethanol, was reduced due to less NADH generated in the fermentation system. Sequential co-fermentation of L. brevis and L. plantarum increased lactic acid yield to 0.80 g g(-1) from poplar hydrolyzate and increased yield to 0.78 g lactic acid per g of biomass from alkali-treated corn stover with minimum by-product formation. Efficient utilization of both cellulose and hemicellulose components of the biomass will improve overall lactic acid production and enable an economical process to produce biodegradable plastics. PMID:25561329

  6. Lactobacillus reuteri-Specific Immunoregulatory Gene rsiR Modulates Histamine Production and Immunomodulation by Lactobacillus reuteri

    PubMed Central

    Hemarajata, P.; Gao, C.; Pflughoeft, K. J.; Thomas, C. M.; Saulnier, D. M.; Spinler, J. K.

    2013-01-01

    Human microbiome-derived strains of Lactobacillus reuteri potently suppress proinflammatory cytokines like human tumor necrosis factor (TNF) by converting the amino acid l-histidine to the biogenic amine histamine. Histamine suppresses mitogen-activated protein (MAP) kinase activation and cytokine production by signaling via histamine receptor type 2 (H2) on myeloid cells. Investigations of the gene expression profiles of immunomodulatory L. reuteri ATCC PTA 6475 highlighted numerous genes that were highly expressed during the stationary phase of growth, when TNF suppression is most potent. One such gene was found to be a regulator of genes involved in histidine-histamine metabolism by this probiotic species. During the course of these studies, this gene was renamed the Lactobacillus reuteri-specific immunoregulatory (rsiR) gene. The rsiR gene is essential for human TNF suppression by L. reuteri and expression of the histidine decarboxylase (hdc) gene cluster on the L. reuteri chromosome. Inactivation of rsiR resulted in diminished TNF suppression in vitro and reduced anti-inflammatory effects in vivo in a trinitrobenzene sulfonic acid (TNBS)-induced mouse model of acute colitis. A L. reuteri strain lacking an intact rsiR gene was unable to suppress colitis and resulted in greater concentrations of serum amyloid A (SAA) in the bloodstream of affected animals. The PhdcAB promoter region targeted by rsiR was defined by reporter gene experiments. These studies support the presence of a regulatory gene, rsiR, which modulates the expression of a gene cluster known to mediate immunoregulation by probiotics at the transcriptional level. These findings may point the way toward new strategies for controlling gene expression in probiotics by dietary interventions or microbiome manipulation. PMID:24123819

  7. Assessment of the in vitro antimicrobial activity of Lactobacillus species for identifying new potential antibiotics.

    PubMed

    Dubourg, Grégory; Elsawi, Ziena; Raoult, Didier

    2015-11-01

    The bacteriocin-mediated antimicrobial properties of Lactobacillus spp. have been widely studied, leading to the use of these micro-organisms in the food industry as preservative agents against foodborne pathogens. In an era in which antibiotic resistance is becoming a public health issue, the antimicrobial activity of Lactobacillus spp. could be used for the discovery of new potential antibiotics. Thus, it is essential to have an accurate method of screening the production of antimicrobial agents by prokaryotes. Many in vitro assays have been published to date, largely concerning but not limited to Lactobacillus spp. However, these methods mainly use the spot-on-the-lawn method, which is prone to contamination during the overlay stage, with protocols using methanol vapours or the reverse side agar technique being applied to avoid such contamination. In this study, a method combining the spot-on-the-lawn and well diffusion methods was tested, permitting clear identification of inhibition zones from eight Lactobacillus spp. towards clinical isolates of 12 species (11 bacteria and 1 yeast) commonly found in human pathology. Lactobacillus plantarum CIP 106786 and Lactobacillus rhamnosus CSUR P567 exhibited the widest antimicrobial activity, whereas Lactobacillus acidophilus strain DSM 20079 was relatively inactive. In addition, the putative MIC50 of L. rhamnosus against Proteus mirabilis was estimated at 1.1×10(9)CFU/mL using culture broth dilution. In conclusion, considering the increasing cultivable bacterial human repertoire, these findings open the way of an effective method to screen in vitro for the production of potential antimicrobial compounds. PMID:26163158

  8. The Probiotic Lactobacillus acidophilus Stimulates Chloride/Hydroxyl Exchange Activity in Human Intestinal Epithelial Cells12

    PubMed Central

    Borthakur, Alip; Gill, Ravinder K.; Tyagi, Sangeeta; Koutsouris, Athanasia; Alrefai, Waddah A.; Hecht, Gail A.; Ramaswamy, Krishnamurthy; Dudeja, Pradeep K.

    2009-01-01

    Probiotics are viable nonpathogenic microorganisms that are considered to confer health benefits to the host. Recent studies indicated that some Lactobacillus species function as probiotics and have been used as alternative treatments for diarrhea, which occurs due to increased secretion, decreased absorption, or both. However, the direct effects of probiotics on intestinal electrolyte absorption are not known. Therefore, we examined the effects of Lactobacillus on luminal chloride/hydroxyl (Cl?/OH?) exchange activity in human intestinal epithelial cells. Postconfluent Caco-2 cells were treated with the Lactobacillus species Lactobacillus acidophilus (LA), Lactobacillus casei, Lactobacillus plantarum, or Lactobacillus rhamnosus (LR) for 3 h at a multiplicity of infection of 50. Cl?/OH? exchange activity was measured as 4,4?-diisothiocyanostilbene-2, 2?-disulfonic acid-sensitive 36Cl uptake in base-loaded cells. Treatment with live, but not heat-killed, LA and LR significantly increased Cl?/OH? exchange activity (~50%), whereas other species were ineffective. Similarly, the conditioned medium (supernatant) of live LA increased Cl?/OH? exchange. The ability of LA or its conditioned culture medium to enhance Cl?/OH? exchange activity was blocked by PI-3 kinase inhibition but was unaffected by inhibition of mitogen-activated protein kinases. Corresponding to the increased Cl?/OH? exchange activity, LA treatment increased the surface expression of the apical anion exchanger, SLC26A3 [Down Regulated in Adenoma (DRA)]. The increased DRA membrane localization might contribute to the increased Cl? absorption by LA. Our results suggest that LA secretes soluble effector molecule(s) into the culture medium that stimulate apical Cl?/OH? exchange activity via phosphatidylinositol-3 kinase mediated mechanism. PMID:18567760

  9. Lactobacillus fermentum ME-3 - an antimicrobial and antioxidative probiotic.

    PubMed

    Mikelsaar, Marika; Zilmer, Mihkel

    2009-04-01

    The paper lays out the short scientific history and characteristics of the new probiotic Lactobacillus fermentum strain ME-3 DSM-14241, elaborated according to the regulations of WHO/FAO (2002). L. fermentum ME-3 is a unique strain of Lactobacillus species, having at the same time the antimicrobial and physiologically effective antioxidative properties and expressing health-promoting characteristics if consumed. Tartu University has patented this strain in Estonia (priority June 2001, patent in 2006), Russia (patent in 2006) and the USA (patent in 2007). The paper describes the process of the identification and molecular typing of this probiotic strain of human origin, its deposition in an international culture collection, and its safety assessment by laboratory tests and testing on experimental animals and volunteers. It has been established that L. fermentum strain ME-3 has double functional properties: antimicrobial activity against intestinal pathogens and high total antioxidative activity (TAA) and total antioxidative status (TAS) of intact cells and lysates, and it is characterized by a complete glutathione system: synthesis, uptake and redox turnover. The functional efficacy of the antimicrobial and antioxidative probiotic has been proven by the eradication of salmonellas and the reduction of liver and spleen granulomas in Salmonella Typhimurium-infected mice treated with the combination of ofloxacin and L. fermentum strain ME-3. Using capsules or foodstuffs enriched with L. fermentum ME-3, different clinical study designs (including double-blind, placebo-controlled, crossover studies) and different subjects (healthy volunteers, allergic patients and those recovering from a stroke), it has been shown that this probiotic increased the antioxidative activity of sera and improved the composition of the low-density lipid particles (LDL) and post-prandial lipids as well as oxidative stress status, thus demonstrating a remarkable anti-atherogenic effect. The elaboration of the probiotic L. fermentum strain ME-3 has drawn on wide international cooperative research and has taken more than 12 years altogether. The new ME-3 probiotic-containing products have been successfully marketed and sold in Baltic countries and Finland. PMID:19381356

  10. Lactobacillus fermentum ME-3 – an antimicrobial and antioxidative probiotic

    PubMed Central

    Mikelsaar, Marika; Zilmer, Mihkel

    2009-01-01

    The paper lays out the short scientific history and characteristics of the new probiotic Lactobacillus fermentum strain ME-3 DSM-14241, elaborated according to the regulations of WHO/FAO (2002). L. fermentum ME-3 is a unique strain of Lactobacillus species, having at the same time the antimicrobial and physiologically effective antioxidative properties and expressing health-promoting characteristics if consumed. Tartu University has patented this strain in Estonia (priority June 2001, patent in 2006), Russia (patent in 2006) and the USA (patent in 2007). The paper describes the process of the identification and molecular typing of this probiotic strain of human origin, its deposition in an international culture collection, and its safety assessment by laboratory tests and testing on experimental animals and volunteers. It has been established that L. fermentum strain ME-3 has double functional properties: antimicrobial activity against intestinal pathogens and high total antioxidative activity (TAA) and total antioxidative status (TAS) of intact cells and lysates, and it is characterized by a complete glutathione system: synthesis, uptake and redox turnover. The functional efficacy of the antimicrobial and antioxidative probiotic has been proven by the eradication of salmonellas and the reduction of liver and spleen granulomas in Salmonella Typhimurium-infected mice treated with the combination of ofloxacin and L. fermentum strain ME-3. Using capsules or foodstuffs enriched with L. fermentum ME-3, different clinical study designs (including double-blind, placebo-controlled, crossover studies) and different subjects (healthy volunteers, allergic patients and those recovering from a stroke), it has been shown that this probiotic increased the antioxidative activity of sera and improved the composition of the low-density lipid particles (LDL) and post-prandial lipids as well as oxidative stress status, thus demonstrating a remarkable anti-atherogenic effect. The elaboration of the probiotic L. fermentum strain ME-3 has drawn on wide international cooperative research and has taken more than 12 years altogether. The new ME-3 probiotic-containing products have been successfully marketed and sold in Baltic countries and Finland. PMID:19381356

  11. Naturally Occurring Lactic Acid Bacteria Isolated from Tomato Pomace Silage

    PubMed Central

    Wu, Jing-jing; Du, Rui-ping; Gao, Min; Sui, Yao-qiang; Xiu, Lei; Wang, Xiao

    2014-01-01

    Silage making has become a significant method of forage conservation worldwide. To determine how tomato pomace (TP) may be used effectively as animal feed, it was ensilaged for 90 days and microbiology counts, fermentation characteristics and chemical composition of tomato pomace silage (TPS) were evaluated at the 30th, 60th, and 90th days, respectively. In addition, 103 lactic acid bacteria were isolated from TPS. Based on the phenotypic and chemotaxonomic characteristics, 16S rDNA sequence and carbohydrate fermentation tests, the isolates were identified as 17 species namely: Lactobacillus coryniformis subsp. torquens (0.97%), Lactobacillus pontis (0.97%), Lactobacillus hilgardii (0.97%), Lactobacillus pantheris (0.97%), Lactobacillus amylovorus (1.9%), Lactobacillus panis (1.9%), Lactobacillus vaginalis (1.9%), Lactobacillus rapi (1.9%), Lactobacillus buchneri (2.9%), Lactobacillus parafarraginis (2.9%), Lactobacillus helveticus (3.9%), Lactobacillus camelliae (3.9%), Lactobacillus fermentum (5.8%), Lactobacillus manihotivorans (6.8%), Lactobacillus plantarum (10.7%), Lactobacillus harbinensis (16.5%) and Lactobacillus paracasei subsp. paracasei (35.0%). This study has shown that TP can be well preserved for 90 days by ensilaging and that TPS is not only rich in essential nutrients, but that physiological and biochemical properties of the isolates could provide a platform for future design of lactic acid bacteria (LAB) inoculants aimed at improving the fermentation quality of silage. PMID:25049999

  12. Screening of immunomodulatory and adhesive Lactobacillus with antagonistic activities against Salmonella from fermented vegetables.

    PubMed

    Feng, Junchang; Liu, Pilong; Yang, Xin; Zhao, Xin

    2015-12-01

    The purpose of this study was to select strains of lactic acid bacteria (LAB) by their in vitro adhesive and immunomodulatory properties for potential use as probiotics. In this study, 16 randomly selected LAB strains from fermented vegetables (sauerkraut, bean and cabbage) were first screened for their tolerance to acid, bile salts, pepsin and pancreatin, bacterial inhibitory activities and abilities to adherence to Caco-2 cells. Then, 4 strains with the highest adhesion abilities were selected for further studies of their immunomodulatory properties and inhibitory effects against Salmonella adhesion and invasion to Caco-2 cells in vitro. The results showed that these 16 LAB strains effectively survived in simulated gastrointestinal condition and inhibited growth of six tested pathogens. Lactobacillus rhamnosus P1, Lactobacillus plantarum P2, Lactobacillus rhamnosus P3 and Lactobacillus casei P4 had the highest abilities to adhere to Caco-2 cells. Furthermore, L. plantarum P2 strain showed higher abilities to induce expression of tumor necrosis factor-? and interleukin-12 by splenic monocytes and strongly inhibited the adhesion and invasion of S. enteritidis ATCC13076 to Caco-2 cells. These results suggest that Lactobacillus strains P2 could be used as a probiotic candidate in food against Salmonella infection. PMID:26340935

  13. Genotyping by randomly amplified polymorphic DNA of bacteriocin producing Lactobacillus acidophilus strains from Nigeria.

    PubMed

    Alli, John Adeolu; Iwalokun, Bamidele A; Oluwadun, Afolabi; Okonko, Iheanyi Omezuruike

    2015-01-01

    Yogurt and starter culture producers are still searching strains of Lactobacillus acidophilus to produce healthier yogurt with a longer shelf life and better texture, taste, and quality. This study determined the genotyping of bacteriocin producing Lactobacillus acidophilus strains recovered from Nigerian yogurts. Yogurt samples were collected from four different states of South West regions of Nigeria. Isolates were obtained from MRS Medium and biochemically characterized. This was further confirmed by API50CH. The bacteriocin positivity and activity was determined. Genomic characterization of our Lactobacillus acidophilus strains was done with randomly amplified polymorphic DNA-PCR. All yogurt samples containing Lactobacillus acidophilus strains meet the probiotic requirement of ?10(6) cfu/mL. The gel picture revealed 6 RAPD clonal types of Lactobacillus acidophilus strains with RAPD type C observed to be more common. Significant differences existed in the mean growth inhibition zone (t = -7.32, P < 0.05 for E. coli ATCC; t = -6.19, P < 0.05 for E. coli clinical isolates; t = -6.16, P < 0.05 for Enterobacter sp; t = -11.92, P < 0.05 for Salmonella typhi, t = -1.10, P > 0.05 Staphylococcus aureus). No correlation between the bacteriocin production, activity, and their RAPD clonal division (X(2) = 7.49, P = 0.1610, df = 5). In conclusion, L. acidophilus isolated in Nigeria samples met the probiotic requirements of ?10(6) cfu/mL and produce bacteriocins with good spectrum of activity. PMID:25153762

  14. Identification of Lactobacillus strains with probiotic features from the bottlenose dolphin (Tursiops truncatus)

    PubMed Central

    Diaz, MA; Bik, EM; Carlin, KP; Venn-Watson, SK; Jensen, ED; Jones, SE; Gaston, EP; Relman, DA; Versalovic, J

    2013-01-01

    Aims In order to develop complementary health management strategies for marine mammals, we used culture-based and culture-independent approaches to identify gastrointestinal lactobacilli of the common bottlenose dolphin, Tursiops truncatus. Methods and Results We screened 307 bacterial isolates from oral and rectal swabs, milk and gastric fluid, collected from 38 dolphins in the U.S. Navy Marine Mammal Program, for potentially beneficial features. We focused our search on lactobacilli and evaluated their ability to modulate TNF secretion by host cells and inhibit growth of pathogens. We recovered Lactobacillus salivarius strains which secreted factors that stimulated TNF production by human monocytoid cells. These Lact. salivarius isolates inhibited growth of selected marine mammal and human bacterial pathogens. In addition, we identified a novel Lactobacillus species by culture and direct sequencing with 96·3% 16S rDNA sequence similarity to Lactobacillus ceti. Conclusions Dolphin-derived Lact. salivarius isolates possess features making them candidate probiotics for clinical studies in marine mammals. Significance and Impact of the Study This is the first study to isolate lactobacilli from dolphins, including a novel Lactobacillus species and a new strain of Lact. salivarius, with potential for veterinary probiotic applications. The isolation and identification of novel Lactobacillus spp. and other indigenous microbes from bottlenose dolphins will enable the study of the biology of symbiotic members of the dolphin microbiota and facilitate the understanding of the microbiomes of these unique animals. PMID:23855505

  15. Selection of potentially probiotic Lactobacillus strains towards their inhibitory activity against poultry enteropathogenic bacteria.

    PubMed

    Kizerwetter-Swida, Magdalena; Binek, Marian

    2005-01-01

    Lactobacilli were isolated from chicken gastrointestinal tract and examined for their potentially probiotic properties towards their inhibitory activity against poultry enteropathogenic bacteria. Biochemical tests, ITS-PCR and cell wall protein analysis were used to characterize the Lactobacillus isolates. The identification of isolated Lactobacillus strains based on phenotypic properties was not always satisfactory. ITS-PCR together with protein profile were found to be helpful in strain identification. Lactobacilli were tested for the inhibitory activity against selected strains of poultry enteropathogenic bacteria (Salmonella Enteritidis, Escherichia coli and Clostridium perfringens). Examined supernatants from Lactobacillus broth cultures demonstrated major antimicrobial activity against C. perfringens. Lower antimicrobial activity were observed against E. coli and Salmonella Enteritidis. The strongest inhibition effect were obtained using supernatant of Lactobacillus acidophilus strain 3D. Results received from this study confirmed that identification of Lactobacillus spp. is often tedious. Some isolates, which are in vitro antagonistic against enteropathogenic bacteria may be considered as potential candidates for poultry probiotics, especially in controlling necrotic enteritis caused by C. perfringens. PMID:16599299

  16. Inerolysin, a Cholesterol-Dependent Cytolysin Produced by Lactobacillus iners?

    PubMed Central

    Rampersaud, Ryan; Planet, Paul J.; Randis, Tara M.; Kulkarni, Ritwij; Aguilar, Jorge L.; Lehrer, Robert I.; Ratner, Adam J.

    2011-01-01

    Lactobacillus iners is a common constituent of the human vaginal microbiota. This species was only recently characterized due to its fastidious growth requirements and has been hypothesized to play a role in the pathogenesis of bacterial vaginosis. Here we present the identification and molecular characterization of a protein toxin produced by L. iners. The L. iners genome encodes an open reading frame with significant primary sequence similarity to intermedilysin (ILY; 69.2% similarity) and vaginolysin (VLY; 68.4% similarity), the cholesterol-dependent cytolysins from Streptococcus intermedius and Gardnerella vaginalis, respectively. Clinical isolates of L. iners produce this protein, inerolysin (INY), during growth in vitro, as assessed by Western analysis. INY is a pore-forming toxin that is activated by reducing agents and inhibited by excess cholesterol. It is active across a pH range of 4.5 to 6.0 but is inactive at pH 7.4. At sublytic concentrations, INY activates p38 mitogen-activated protein kinase and allows entry of fluorescent phalloidin into the cytoplasm of epithelial cells. Unlike VLY and ILY, which are human specific, INY is active against cells from a broad range of species. INY represents a new target for studies directed at understanding the role of L. iners in states of health and disease at the vaginal mucosal surface. PMID:21169489

  17. Kinetics and Modeling of Lactic Acid Production by Lactobacillus plantarum

    PubMed Central

    Passos, Frederico V.; Fleming, Henry P.; Ollis, David F.; Felder, Richard M.; McFeeters, R. F.

    1994-01-01

    An unstructured model was developed to describe bacterial growth, substrate utilization, and lactic acid production by Lactobacillus plantarum in cucumber juice. Significant lactic acid production occurred during growth, as well as stationary phases. The percentage of acid produced after growth ceased was a function of the medium composition. Up to 51% of the lactic acid was produced after growth ceased when NaCl was not present in the medium, whereas not more than 18% of the total lactic acid was produced after the growth ceased in presence of NaCl, probably because of an increase in the cell death rate. An equation relating the specific death rate and NaCl concentration was developed. With the kinetic model proposed by R. Luedeking and E. L. Piret (J. Biochem. Microbiol. Technol. Eng. 1:393-412, 1958) for lactic acid production rate, the growth-associated and non-growth-associated coefficients were determined as 51.9 (±4.2) mmol/g of cells and 7.2 (±0.9) mmol/g of cells h-1 respectively. The model was demonstrated for batch growth of L. plantarum in cucumber juice. Mathematical simulations were used to predict the influence of variations in death rate, proton concentration when growth ceased, and buffer capacity of the juice on the overall fermentation process. PMID:16349339

  18. Acid Tolerance of Leuconostoc mesenteroides and Lactobacillus plantarum†

    PubMed Central

    McDonald, L. C.; Fleming, H. P.; Hassan, H. M.

    1990-01-01

    In this study, we determined the internal cellular pH response of Leuconostoc mesenteroides and Lactobacillus plantarum to the external pH created by the microorganisms themselves or by lactic or acetic acids and their salts added to the growth medium. Growth of Leuconostoc mesenteroides stopped when its internal pH reached 5.4 to 5.7, and growth of L. plantarum stopped when its internal pH reached 4.6 to 4.8. Variation in growth medium composition or pH did not alter the growth-limiting internal pH reached by these microorganisms. L. plantarum maintained its pH gradient in the presence of either 160 mM sodium acetate or sodium lactate down to an external pH of 3.0 with either acid. In contrast, the ?pH of Leuconostoc mesenteroides was zero at pH 4.0 with acetate and 5.0 with lactate. No differences were found between d-(?)- and l-(+)-lactic acid for the limiting internal pH for growth of either microorganism. The comparatively low growth-limiting internal pH and ability to maintain a pH gradient at high organic acid concentration may contribute to the ability of L. plantarum to terminate vegetable fermentations. PMID:16348238

  19. The Adsorption of Ochratoxin A by Lactobacillus Species

    PubMed Central

    Piotrowska, Ma?gorzata

    2014-01-01

    The objective of this study was to examine ochratoxin A (OTA) binding by three lactic acid bacteria (LAB) species: Lactobacillus plantarum, L. brevis, and L. sanfranciscensis. Experiments were conducted using MRS medium and PBS buffer contaminated with 1000 ng/mL OTA and inoculated with live or thermally inactivated bacterial biomass at a concentration of 1 or 5 mg dry weight/mL. It was found that, depending on the strain and biomass density, live bacterial cells reduced OTA content by 16.9% to 35% in MRS medium and by 14.8% to 26.4% in PBS after 24 h of contact. OTA binding was higher in the case of thermally inactivated bacterial biomass (46.2% to 59.8%). The process is very rapid: OTA was removed from PBS as early as after 30 min of contact. The binding of the toxin by cells was partially reversible under the treatment by water and 1 M HCl. The results show that OTA is adsorbed to the surface structures of the cell wall, which is promoted not only by the hydrophobic properties of the cell wall, but also by electron donor-acceptor and Lewis acid-base interactions. PMID:25247265

  20. The adsorption of ochratoxin a by lactobacillus species.

    PubMed

    Piotrowska, Ma?gorzata

    2014-09-01

    The objective of this study was to examine ochratoxin A (OTA) binding by three lactic acid bacteria (LAB) species: Lactobacillus plantarum, L. brevis, and L. sanfranciscensis. Experiments were conducted using MRS medium and PBS buffer contaminated with 1000 ng/mL OTA and inoculated with live or thermally inactivated bacterial biomass at a concentration of 1 or 5 mg dry weight/mL. It was found that, depending on the strain and biomass density, live bacterial cells reduced OTA content by 16.9% to 35% in MRS medium and by 14.8% to 26.4% in PBS after 24 h of contact. OTA binding was higher in the case of thermally inactivated bacterial biomass (46.2% to 59.8%). The process is very rapid: OTA was removed from PBS as early as after 30 min of contact. The binding of the toxin by cells was partially reversible under the treatment by water and 1 M HCl. The results show that OTA is adsorbed to the surface structures of the cell wall, which is promoted not only by the hydrophobic properties of the cell wall, but also by electron donor-acceptor and Lewis acid-base interactions. PMID:25247265

  1. Lactobacillus acidophilus—Rutin Interplay Investigated by Proteomics

    PubMed Central

    Mazzeo, Maria Fiorella; Lippolis, Rosa; Sorrentino, Alida; Liberti, Sarah; Fragnito, Federica; Siciliano, Rosa Anna

    2015-01-01

    Dietary polyphenols are bioactive molecules that beneficially affect human health, due to their anti-oxidant, anti-inflammatory, cardio-protective and chemopreventive properties. They are absorbed in a very low percentage in the small intestine and reach intact the colon, where they are metabolized by the gut microbiota. Although it is well documented a key role of microbial metabolism in the absorption of polyphenols and modulation of their biological activity, molecular mechanisms at the basis of the bacteria-polyphenols interplay are still poorly understood. In this context, differential proteomics was applied to reveal adaptive response mechanisms that enabled a potential probiotic Lactobacillus acidophilus strain to survive in the presence of the dietary polyphenol rutin. The response to rutin mainly modulated the expression level of proteins involved in general stress response mechanisms and, in particular, induced the activation of protein quality control systems, and affected carbohydrate and amino acid metabolism, protein synthesis and cell wall integrity. Moreover, rutin triggered the expression of proteins involved in oxidation-reduction processes.This study provides a first general view of the impact of dietary polyphenols on metabolic and biological processes of L. acidophilus. PMID:26544973

  2. Enteric coating of granules containing the probiotic Lactobacillus acidophilus.

    PubMed

    Pyar, Hassan; Peh, Kok-Khiang

    2014-06-01

    In the present study, a capsule formulation composed of enteric coated granules of Lactobacillus acidophilus ATCC 4962 was developed using Eudragit L30D-55 as enteric polymer. Optimization of the capsule formulation was achieved with a maximum viable cell count after 2 h of incubation in acid medium and disintegration time of 1 h in buffer pH 6.8. The amount of Eudragit L30D-55 in the capsules correlated with gastric juice resistance. The best protective qualities against artificial gastric juice were observed when capsules were prepared from granules composed of L. acidophilus, corn starch, lactose monohydrate, polyvinylpyrrolidone and coated with 12.5 % (m/V) of Eudragit L30D-55. Capsule formulation of L. acidophilus in edible broth medium suspension serves as a cheap alternative to the expensive freeze-drying procedure for preparing L. acidophilus. In addition, the enteric coating using Eudragit L30D-55 could protect probiotics from the acidic gastric environment and enhance the bioactivity of probiotics along with replacement of pathogenic microbes in human intestine. PMID:24914724

  3. Timely approaches to identify probiotic species of the genus Lactobacillus

    PubMed Central

    2013-01-01

    Over the past decades the use of probiotics in food has increased largely due to the manufacturer’s interest in placing “healthy” food on the market based on the consumer’s ambitions to live healthy. Due to this trend, health benefits of products containing probiotic strains such as lactobacilli are promoted and probiotic strains have been established in many different products with their numbers increasing steadily. Probiotics are used as starter cultures in dairy products such as cheese or yoghurts and in addition they are also utilized in non-dairy products such as fermented vegetables, fermented meat and pharmaceuticals, thereby, covering a large variety of products. To assure quality management, several pheno-, physico- and genotyping methods have been established to unambiguously identify probiotic lactobacilli. These methods are often specific enough to identify the probiotic strains at genus and species levels. However, the probiotic ability is often strain dependent and it is impossible to distinguish strains by basic microbiological methods. Therefore, this review aims to critically summarize and evaluate conventional identification methods for the genus Lactobacillus, complemented by techniques that are currently being developed. PMID:24063519

  4. Towards a better understanding of Lactobacillus rhamnosus GG - host interactions

    PubMed Central

    2014-01-01

    Lactobacillus rhamnosus GG (LGG) is one of the most widely used probiotic strains. Various health effects are well documented including the prevention and treatment of gastro-intestinal infections and diarrhea, and stimulation of immune responses that promote vaccination or even prevent certain allergic symptoms. However, not all intervention studies could show a clinical benefit and even for the same conditions, the results are not univocal. Clearly, the host phenotype governed by age, genetics and environmental factors such as the endogenous microbiota, plays a role in whether individuals are responders or non-responders. However, we believe that a detailed knowledge of the bacterial physiology and the LGG molecules that play a key role in its host-interaction capacity is crucial for a better understanding of its potential health benefits. Molecules that were yet identified as important factors governing host interactions include its adhesive pili or fimbriae, its lipoteichoic acid molecules, its major secreted proteins and its galactose-rich exopolysaccharides, as well as specific DNA motifs. Nevertheless, future studies are needed to correlate specific health effects to these molecular effectors in LGG, and also in other probiotic strains. PMID:25186587

  5. Human ?-amylase Present in Lower-Genital-Tract Mucosal Fluid Processes Glycogen to Support Vaginal Colonization by Lactobacillus

    PubMed Central

    Spear, Gregory T.; French, Audrey L.; Gilbert, Douglas; Zariffard, M. Reza; Mirmonsef, Paria; Sullivan, Thomas H.; Spear, William W.; Landay, Alan; Micci, Sandra; Lee, Byung-Hoo; Hamaker, Bruce R.

    2014-01-01

    Lactobacillus colonization of the lower female genital tract provides protection from the acquisition of sexually transmitted diseases, including human immunodeficiency virus, and from adverse pregnancy outcomes. While glycogen in vaginal epithelium is thought to support Lactobacillus colonization in vivo, many Lactobacillus isolates cannot utilize glycogen in vitro. This study investigated how glycogen could be utilized by vaginal lactobacilli in the genital tract. Several Lactobacillus isolates were confirmed to not grow in glycogen, but did grow in glycogen-breakdown products, including maltose, maltotriose, maltopentaose, maltodextrins, and glycogen treated with salivary ?-amylase. A temperature-dependent glycogen-degrading activity was detected in genital fluids that correlated with levels of ?-amylase. Treatment of glycogen with genital fluids resulted in production of maltose, maltotriose, and maltotetraose, the major products of ?-amylase digestion. These studies show that human ?-amylase is present in the female lower genital tract and elucidates how epithelial glycogen can support Lactobacillus colonization in the genital tract. PMID:24737800

  6. Characterization of the most abundant Lactobacillus species in chicken gastrointestinal tract and potential use as probiotics for genetic engineering.

    PubMed

    Wang, Lei; Fang, Mingjian; Hu, Yanping; Yang, Yuxin; Yang, Mingming; Chen, Yulin

    2014-07-01

    The count and diffusion of Lactobacilli species in the different gastrointestinal tract (GI) regions of broilers were investigated by quantitative real-time polymerase chain reaction, and the probiotic characteristics of six L. reuteri species isolated from broilers' GI tract were also investigated to obtain the potential target for genetic engineering. Lactobacilli had the highest diversity in the crop and the lowest one in the cecum. Compared with the lower GI tract, more Lactobacilli were found in the upper GI tract. Lactobacillus reuteri, L. johnsonii, L. acidophilus, L. crispatus, L. salivarius, and L. aviarius were the predominant Lactobacillus species and present throughout the GI tract of chickens. Lactobacillus reuteri was the most abundant Lactobacillus species. Lactobacillus reuteri XC1 had good probiotic characteristics that would be a potential and desirable target for genetic engineering. PMID:24850302

  7. Bacteriocins from Lactobacillus plantarum – production, genetic organization and mode of action

    PubMed Central

    Todorov, Svetoslav D.

    2009-01-01

    Bacteriocins are biologically active proteins or protein complexes that display a bactericidal mode of action towards usually closely related species. Numerous strains of bacteriocin producing Lactobacillus plantarum have been isolated in the last two decades from different ecological niches including meat, fish, fruits, vegetables, and milk and cereal products. Several of these plantaricins have been characterized and the aminoacid sequence determined. Different aspects of the mode of action, fermentation optimization and genetic organization of the bacteriocin operon have been studied. However, numerous of bacteriocins produced by different Lactobacillus plantarum strains have not been fully characterized. In this article, a brief overview of the classification, genetics, characterization, including mode of action and production optimization for bacteriocins from Lactic Acid Bacteria in general, and where appropriate, with focus on bacteriocins produced by Lactobacillus plantarum, is presented. PMID:24031346

  8. Lactobacillus and Pediococcus species richness and relative abundance in the vagina of rhesus monkeys (Macaca mulatta)

    PubMed Central

    Gravett, Michael G.; Jin, Ling; Pavlova, Sylvia I.; Tao, Lin

    2012-01-01

    Background The rhesus monkey is an important animal model to study human vaginal health to which lactic acid bacteria play a significant role. However, the vaginal lactic acid bacterial species richness and relative abundance in rhesus monkeys is largely unknown. Methods Vaginal swab samples were aseptically obtained from 200 reproductive aged female rhesus monkeys. Following Rogosa agar plating, single bacterial colonies representing different morphotypes were isolated and analyzed for whole-cell protein profile, species-specifc PCR, and 16S rRNA gene sequence. Results A total of 510 Lactobacillus strains of 17 species and one Pediococcus acidilactici were identified. The most abundant species was L. reuteri, which colonized the vaginas of 86% monkeys. L. johnsonii was the second most abundant species, which colonized 36% of monkeys. The majority of monkeys were colonized by multiple Lactobacillus species. Conclusions The vaginas of rhesus monkeys are frequently colonized by multiple Lactobacillus species, dominated by L. reuteri. PMID:22429090

  9. Isolation and microencapsulation of Lactobacillus spp. from corn silage for probiotic application

    PubMed Central

    Kasra – Kermanshahi, R; Fooladi, J; Peymanfar, S

    2010-01-01

    Background and Objectives Probiotics including strains of Lactobacillus spp. are living microorganisms including which are beneficial to human and animals health. In this study, Lactobacillus has been isolated from corn silage in a cold region of Iran by anaerobic culture. Materials and Methods The bacteriological and biochemical standard methods were used for identification and phenotypic characterization of isolated organism. To increase the stability of organism in the environment, we used microencapsulation technique using stabilizer polymers (Alginate and Chitosan). Results The isolated Lactobacillus spp. was able to ferment tested carbohydrates and grow at 10°C–50°C. Using microencapsulation, the stability and survival of this bacterium increased. Conclusion microencapsulation of lactic acid bacteria with alginate and chitosan coating offers an effective way of delivering viable bacterial cells to the colon and maintaining their survival during refrigerated storage. PMID:22347557

  10. Lactobacillus-mediated priming of the respiratory mucosa protects against lethal pneumovirus infection*

    PubMed Central

    Gabryszewski, Stanislaw J.; Bachar, Ofir; Dyer, Kimberly D.; Percopo, Caroline M.; Killoran, Kristin E.; Domachowske, Joseph B.; Rosenberg, Helene F.

    2012-01-01

    The inflammatory response to respiratory virus infection can be complex and refractory to standard therapy. Lactobacilli, when targeted to the respiratory epithelium, are highly effective at suppressing virus-induced inflammation and protecting against lethal disease. Specifically, wild-type mice primed via intranasal inoculation with live or heat-inactivated Lactobacillus plantarum or Lactobacillus reuteri were completely protected against lethal infection with the virulent rodent pathogen, pneumonia virus of mice (PVM); significant protection (60% survival) persisted for at least thirteen weeks. Protection was not unique to Lactobacillus species, and was also observed in response to priming with non-pathogenic gram-positive Listeria innocua. Priming with live lactobacilli resulted in diminished granulocyte recruitment, diminished expression of multiple proinflammatory cytokines (CXCL10, CXCL1, CCL2, and TNF) and reduced virus recovery, although we have demonstrated clearly that absolute virus titer does not predict clinical outcome. Lactobacillus priming also resulted in prolonged survival and protection against the lethal sequelae of PVM infection in MyD88 gene-deleted (MyD88?/?) mice, suggesting that the protective mechanisms may be Toll-like receptor-independent. Most intriguing, virus recovery and cytokine expression patterns in Lactobacillus-primed MyD88?/? mice were indistinguishable from those observed in control-primed MyD88?/? counterparts, In summary, we have identified and characterized an effective Lactobacillus-mediated innate immune shield, which may ultimately serve as critical and long-term protection against infection in the absence of specific antiviral vaccines. PMID:21169550

  11. Evaluation of Phytate-Degrading Lactobacillus Culture Administration to Broiler Chickens

    PubMed Central

    Askelson, Tyler E.; Campasino, Ashley; Lee, Jason T.

    2014-01-01

    Probiotics have been demonstrated to promote growth, stimulate immune responses, and improve food safety of poultry. While widely used, their effectiveness is mixed, and the mechanisms through which they contribute to poultry production are not well understood. Microbial phytases are increasingly supplemented in feed to improve digestibility and reduce antinutritive effects of phytate. The microbial origin of these exogenous enzymes suggests a potentially important mechanism of probiotic functionality. We investigated phytate degradation as a novel probiotic mechanism using recombinant Lactobacillus cultures expressing Bacillus subtilis phytase. B. subtilis phyA was codon optimized for expression in Lactobacillus and cloned into the expression vector pTRK882. The resulting plasmid, pTD003, was transformed into Lactobacillus acidophilus, Lactobacillus gallinarum, and Lactobacillus gasseri. SDS-PAGE revealed a protein in the culture supernatants of Lactobacillus pTD003 transformants with a molecular weight similar to that of the B. subtilis phytase. Expression of B. subtilis phytase increased phytate degradation of L. acidophilus, L. gasseri, and L. gallinarum approximately 4-, 10-, and 18-fold over the background activity of empty-vector transformants, respectively. Phytase-expressing L. gallinarum and L. gasseri were administered to broiler chicks fed a phosphorus-deficient diet. Phytase-expressing L. gasseri improved weight gain of broiler chickens to a level comparable to that for chickens fed a control diet adequate in phosphorus, demonstrating proof of principle that administration of phytate-degrading probiotic cultures can improve performance of livestock animals. This will inform future studies investigating whether probiotic cultures are able to provide both the performance benefits of feed enzymes and the animal health and food safety benefits traditionally associated with probiotics. PMID:24271165

  12. Association between Obesity and Cervical Microflora Dominated by Lactobacillus iners in Korean Women.

    PubMed

    Oh, Hea Young; Seo, Sang-Soo; Kong, Ji-Sook; Lee, Jae-Kwan; Kim, Mi Kyung

    2015-10-01

    Lactobacillus spp. are associated with the maintenance of reproductive health, but obesity reduces fertility and is a risk factor for obstetric and neonatal complications. We assessed the association between obesity and the cervical Lactobacillus composition, which has not been examined previously. Pyrosequencing was performed using cervical swabs collected from 76 normal participants with negative results for cervical intraepithelial neoplasia (CIN) and 57 participants with CIN, based on histological examinations. Cluster analysis of nine Lactobacillus spp. was performed, and five cluster types were identified. The association between obesity and the Lactobacillus community was assessed by logistic regression analysis after adjustment for confounding factors. The proportion of Lactobacillus iners increased and that of Lactobacillus crispatus decreased according to body mass index (BMI) categories, i.e., underweight (BMI of <18.5 kg m(-2)), normal weight (BMI of 18.5 to 22.9 kg m(-2)), overweight (BMI of 23.0 to 24.9 kg m(-2)), and obese (BMI of ?25 kg m(-2)). The L. iners-dominant type had a significant association with obesity (odds ratio [OR], 7.55 [95% confidence interval [CI], 1.18 to 48.2]), compared to the L. crispatus-dominant type. The group with high values for the ratio obtained by dividing the relative abundance of L. iners by that of L. crispatus had a significant association with obesity (OR, 6.54 [95% CI, 1.22 to 35.1]), compared to the low-ratio group. Associations between obesity and the L. iners/L. crispatus ratio were observed among young women (OR, 6.26 [95% CI, 1.15 to 33.9]) but not older women and in the normal group (OR, 6.97 [95% CI, 1.20 to 70.4]) but not the CIN group. Obesity was associated with cervical microflora dominated by L. iners in reproductive-age women without dysplasia. PMID:26269625

  13. Lactobacillus brantae sp. nov., isolated from faeces of Canada geese (Branta canadensis).

    PubMed

    Volokhov, Dmitriy V; Amselle, Megan; Beck, Brian J; Popham, David L; Whittaker, Paul; Wang, Hua; Kerrigan, Elizabeth; Chizhikov, Vladimir E

    2012-09-01

    Three strains of lactic acid bacteria (LAB) were isolated from the faeces of apparently healthy wild Canada geese (Branta canadensis) in 2010 by cultivating faecal LAB on Rogosa SL agar under aerobic conditions. These three isolates were found to share 99.9 % gene sequence similarity of their 16S rRNA, their 16S-23S intergenic transcribed spacer region (ITS), partial 23S rRNA, rpoB, rpoC, rpoA and pheS gene sequences. However, the three strains exhibited lower levels of sequence similarity of these genetic targets to all known LAB, and the phylogenetically closest species to the geese strains were Lactobacillus casei, Lactobacillus paracasei, Lactobacillus rhamnosus and Lactobacillus saniviri. In comparison to L. casei ATCC 393(T), L. paracasei ATCC 25302(T), L. rhamnosus ATCC 7469(T) and L. saniviri DSM 24301(T), the novel isolates reacted uniquely in tests for cellobiose, galactose, mannitol, citric acid, aesculin and dextrin, and gave negative results in tests for l-proline arylamidase and l-pyrrolydonyl-arylamidase, and in the Voges-Proskauer test. Biochemical tests for cellobiose, aesculin, galactose, gentiobiose, mannitol, melezitose, ribose, salicin, sucrose, trehalose, raffinose, turanose, amygdalin and arbutin could be used for differentiation between L. saniviri and the novel strains. On the basis of phenotypic and genotypic characteristics, and phylogenetic data, the three isolates represent a novel species of the genus Lactobacillus, for which the name Lactobacillus brantae sp. nov. is proposed. The type strain is SL1108(T) (= ATCC BAA-2142(T) = LMG 26001(T) = DSM 23927(T)) and two additional strains are SL1170 and SL60106. PMID:22021580

  14. Free Glycogen in Vaginal Fluids Is Associated with Lactobacillus Colonization and Low Vaginal pH

    PubMed Central

    Mirmonsef, Paria; Hotton, Anna L.; Gilbert, Douglas; Burgad, Derick; Landay, Alan; Weber, Kathleen M.; Cohen, Mardge; Ravel, Jacques; Spear, Gregory T.

    2014-01-01

    Objective Lactobacillus dominates the lower genital tract microbiota of many women, producing a low vaginal pH, and is important for healthy pregnancy outcomes and protection against several sexually transmitted pathogens. Yet, factors that promote Lactobacillus remain poorly understood. We hypothesized that the amount of free glycogen in the lumen of the lower genital tract is an important determinant of Lactobacillus colonization and a low vaginal pH. Methods Free glycogen in lavage samples was quantified. Pyrosequencing of the 16S rRNA gene was used to identify microbiota from 21 African American women collected over 8–11 years. Results Free glycogen levels varied greatly between women and even in the same woman. Samples with the highest free glycogen had a corresponding median genital pH that was significantly lower (pH 4.4) than those with low glycogen (pH 5.8; p<0.001). The fraction of the microbiota consisting of Lactobacillus was highest in samples with high glycogen versus those with low glycogen (median?=?0.97 vs. 0.05, p<0.001). In multivariable analysis, having 1 vs. 0 male sexual partner in the past 6 months was negatively associated, while BMI ?30 was positively associated with glycogen. High concentrations of glycogen corresponded to higher levels of L. crispatus and L. jensenii, but not L. iners. Conclusion These findings show that free glycogen in genital fluid is associated with a genital microbiota dominated by Lactobacillus, suggesting glycogen is important for maintaining genital health. Treatments aimed at increasing genital free glycogen might impact Lactobacillus colonization. PMID:25033265

  15. Oral immunization with recombinant lactobacillus plantarum induces a protective immune response in mice with Lyme disease.

    PubMed

    del Rio, Beatriz; Dattwyler, Raymond J; Aroso, Miguel; Neves, Vera; Meirelles, Luciana; Seegers, Jos F M L; Gomes-Solecki, Maria

    2008-09-01

    Mucosal immunization is advantageous over other routes of antigen delivery because it can induce both mucosal and systemic immune responses. Our goal was to develop a mucosal delivery vehicle based on bacteria generally regarded as safe, such as Lactobacillus spp. In this study, we used the Lyme disease mouse model as a proof of concept. We demonstrate that an oral vaccine based on live recombinant Lactobacillus plantarum protects mice from tick-transmitted Borrelia burgdorferi infection. Our method of expressing vaccine antigens in L. plantarum induces both systemic and mucosal immunity after oral administration. This platform technology can be applied to design oral vaccine delivery vehicles against several microbial pathogens. PMID:18632920

  16. Probiotics in digestive diseases: focus on Lactobacillus GG.

    PubMed

    Pace, F; Pace, M; Quartarone, G

    2015-12-01

    Probiotics are becoming increasingly important in basic and clinical research, but they are also a subject of considerable economic interest due to their expanding popularity. They are live micro-organisms which, when administered in adequate amounts, confer a health benefit to the host. From this very well-known definition, it is clear that, unlike drugs, probiotics might be useful in healthy subjects to reduce the risk of developing certain diseases or to optimise some physiological functions. They also may offer some advantages in already ill persons in relieving symptoms and signs, e.g. people with acute diarrhea. According to current definitions, probiotics should survive both gastric acid and bile to reach the small intestine and colon, where they exert their effects. Many of these are available in a lyophilized (freeze-dried) pill form, though some are available in yogurt or as packets (sachets), which can be mixed into non-carbonated drinks. The present review focuses on three main issues: 1) understanding why, at present, probiotics are so interesting for doctors and consumers; 2) reviewing the available data on probiotic use in digestive diseases, in particular irritable bowel syndrome (IBS), (prevention of) infectious diarrhea, inflammatory bowel disease (IBD), non-alcoholic fatty liver disease (NAFLD), and colorectal cancer (CRC); 3) highlighting the individual profile of Lactobacillus GG (LGG) in the above contexts, providing an assessment as well as recommendations on its use in gastro-intestinal tract (GIT) disorders. Research studies conducted in animals and humans with the main probiotics strains for GIT diseases, and published from the early 1990s to 2014 have been considered. PubMed, Medline and Ovid were the main sources adopted for data retrieving. The increasing attention on probiotics is a direct consequence of the improvement in the techniques for studying microbiota. Until recently, its composition has been analysed by culture-based methods that use differential media to select for specific populations of bacteria according to their metabolic requirements. Lactobacillus and Bifidobacterium species are by and large the most commonly used probiotics. Strictly speaking, however, the term "probiotic" should be reserved for live microbes that have been shown in controlled human studies to provide a health benefit. Taking into account patients suffering from the most common gastrointestinal diseases, in whose establishment the GI microbiota plays a key role, probiotics have to be considered as very promising agents, capable of beneficially modulating the intestinal ecosystem, which is perturbed in cases of dysbiosis. Although more clinical data are still needed to better assess the clinical relevance of probiotics, to date, procariota such as Bifidobacteria and Lactobacilli strains, and eucariota such as some Saccharomyces strains are among the most widely used agents in GIT disorders. LGG is a well-known probiotic strain that was isolated more than 20 years ago by Goldin and Gorbach from a faecal sample of a healthy adult, based on several selection criteria: high adhesion in vitro, high resistance against gastric acidity and high antimicrobial activity against pathogens such as Salmonella. In vivo studies have also shown a good persistence of LGG in the human GIT. Since its isolation, LGG has become one of the best clinically documented probiotic strains. A growing body of evidence suggests benefits such as prevention and relief of various types of diarrhoea, and treatment of relapsing Clostridium difficile colitis. Thus, with respect to both adaptation to the GIT and probiotic effects, LGG can be regarded as a prototypical probiotic strain. PMID:26657927

  17. Uracil salvage pathway in Lactobacillus plantarum: Transcription and genetic studies.

    PubMed

    Arsène-Ploetze, Florence; Nicoloff, Hervé; Kammerer, Benoît; Martinussen, Jan; Bringel, Françoise

    2006-07-01

    The uracil salvage pathway in Lactobacillus plantarum was demonstrated to be dependent on the upp-pyrP gene cluster. PyrP was the only high-affinity uracil transporter since a pyrP mutant no longer incorporated low concentrations of radioactively labeled uracil and had increased resistance to the toxic uracil analogue 5-fluorouracil. The upp gene encoded a uracil phosphoribosyltransferase (UPRT) enzyme catalyzing the conversion of uracil and 5-phosphoribosyl-alpha-1-pyrophosphate to UMP and pyrophosphate. Analysis of mutants revealed that UPRT is a major cell supplier of UMP synthesized from uracil provided by preformed nucleic acid degradation. In a mutant selection study, seven independent upp mutants were isolated and all were found to excrete low amounts of pyrimidines to the growth medium. Pyrimidine-dependent transcription regulation of the biosynthetic pyrimidine pyrR1-B-C-Aa1-Ab1-D-F-E operon was impaired in the upp mutants. Despite the fact that upp and pyrP are positioned next to each other on the chromosome, they are not cotranscribed. Whereas pyrP is expressed as a monocistronic message, the upp gene is part of the lp_2376-glyA-upp operon. The lp_2376 gene encodes a putative protein that belongs to the conserved protein family of translation modulators such as Sua5, YciO, and YrdC. The glyA gene encodes a putative hydroxymethyltransferase involved in C1 unit charging of tetrahydrofolate, which is required in the biosynthesis of thymidylate, pantothenate, and purines. Unlike upp transcription, pyrP transcription is regulated by exogenous pyrimidine availability, most likely by the same mechanism of transcription attenuation as that of the pyr operon. PMID:16788187

  18. Resistance of functional Lactobacillus plantarum strains against food stress conditions.

    PubMed

    Ferrando, Verónica; Quiberoni, Andrea; Reinhemer, Jorge; Suárez, Viviana

    2015-06-01

    The survival of three Lactobacillus plantarum strains (Lp 790, Lp 813 and Lp 998) with functional properties was studied taking into account their resistance to thermal, osmotic and oxidative stress factors. Stress treatments applied were: 52 °C-15 min (Phosphate Buffer pH 7, thermal shock), H2O2 0.1% (p/v) - 30 min (oxidative shock) and NaCl aqueous solution at 17, 25 and 30% (p/v) (room temperature - 1 h, osmotic shock). The osmotic stress was also evaluated on cell growth in MRS broth added of 2, 4, 6, 8 and 10% (p/v) of NaCl, during 20 h at 30 °C. The cell thermal adaptation was performed in MRS broth, selecting 45 °C for 30 min as final conditions for all strains. Two strains (Lp 813 and Lp 998) showed, in general, similar behaviour against the three stress factors, being clearly more resistant than Lp 790. An evident difference in growth kinetics in presence of NaCl was observed between Lp 998 and Lp 813, Lp998 showing a higher optical density (OD570nm) than Lp 813 at the end of the assay. Selected thermal adaptation improved by 2 log orders the thermal resistance of both strains, but cell growth in presence of NaCl was enhanced only in Lp 813. Oxidative resistance was not affected with this thermal pre-treatment. These results demonstrate the relevance of cell technological resistance when selecting presumptive "probiotic" cultures, since different stress factors might considerably affect viability or/and performance of the strains. The incidence of stress conditions on functional properties of the strains used in this work are currently under research in our group. PMID:25790993

  19. Mixed culture models for predicting intestinal microbial interactions between Escherichia coli and Lactobacillus in the presence of probiotic Bacillus subtilis.

    PubMed

    Yang, J J; Niu, C C; Guo, X H

    2015-12-01

    Bacillus has been proposed as a probiotic due to its in vivo effectiveness in the gastrointestinal tract through antimicrobial activities. The present study investigates the effects of Lactobacillus alone or in the presence of Bacillus subtilis MA139 on the inhibition of pathogenic Escherichia coli K88. Mixed cultures were used to predict the possible interactions among these bacteria within the intestinal tract of animals. B. subtilis MA139 was first assayed for its inhibition against E. coli K88 both under shaking and static culture conditions. A co-culture assay was employed under static conditions to test the inhibitory effects of Lactobacillus reuteri on E. coli K88, with or without addition of B. subtilis MA139. The results showed that B. subtilis MA139 had marked inhibition against E. coli K88 under shaking conditions and weak inhibition under static conditions. Lactobacillus alone as well as in combination with B. subtilis MA139 spores exerted strong inhibition against E. coli K88 under static conditions. However, the inhibition by Lactobacillus in combination with B. subilis spores was much higher than that by Lactobacillus alone (P<0.01). B. subtilis MA139 significantly decreased the pH and oxidation-reduction potential values of the co-culture broth compared to that of Lactobacillus alone (P<0.05). The viability of Lactobacillus increased when co-cultured with B. subtilis MA139 because of significantly higher Lactobacillus counts and lower pH values in the broth (P<0.05). The role of Bacillus in the mixed culture models suggests that Bacillus may produce beneficial effects by increasing the viability of lactobacilli and subsequently inhibiting the growth of pathogenic E. coli. Therefore, the combination of Bacillus and Lactobacillus species as a probiotic is recommended. PMID:26259891

  20. Effect of malic acid on the growth kinetics of Lactobacillus plantarum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fermentation kinetics of Lactobacillus plantarum was studied in a specially designed broth formulated from commercially available, dehydrated components (YTA - yeast extract, trypticase, and ammonium sulfate) in batch and continuous culture. During batch growth in the absence of malic acid in t...

  1. Draft Genome Sequence of the Mannitol-Producing Strain Lactobacillus mucosae CRL573

    PubMed Central

    Bleckwedel, Juliana; Terán, Lucrecia C.; Bonacina, Julieta; Saavedra, Lucila

    2014-01-01

    Lactobacillus mucosae CRL573, isolated from child fecal samples, efficiently converts fructose and/or sucrose into the low-calorie sugar mannitol when cultured in modified MRS medium at pH 5.0. Also, the strain is capable of producing bacteriocin. The draft genome sequence of this strain with potential industrial applications is presented here. PMID:25502678

  2. In vitro conjugal transfer of tetracycline resistance from Lactobacillus isolates to other Gram-positive bacteria

    E-print Network

    Gent, Universiteit

    be spread to other lactic acid bacteria. In order to assess the risk of this potential hazard, the magnitude from fermented dry sausages, to transfer tetracycline resistance encoded by tet(M) through conjugation alimentarius; Lactobacillus sakei subsp. sakei; Fermented dry sausage 1. Introduction Lactobacilli are common

  3. Lactobacillus amylovorus, a new starch-hydrolyzing species from cattle waste-corn fermentations

    SciTech Connect

    Nakamura, L.K.

    1981-01-01

    The morphology, physiology and fermentation characteristics of this hitherto unrecognized species are described. The new Lactobacillus species can be differentiated from L. acidophilus, L. jensenii, and L. leichmannii on the basis of starch fermentation, G + C content, vitamin requirements and stereoisomerism of lactic acid produced. The type strain of L. amylovorus is NRRL B-4540. (Refs. 39).

  4. Mutation and Selection of Lactobacillus plantarum Strains That Do Not Produce Carbon Dioxide from Malate †

    PubMed Central

    Daeschel, M. A.; McFeeters, R. F.; Fleming, H. P.; Klaenhammer, T. R.; Sanozky, R. B.

    1984-01-01

    A differential medium was developed to distinguish between malate-decarboxylating (MDC+) and -non-decarboxylating (MDC?) strains of Lactobacillus plantarum. MDC? strains produced a visible acid reaction in the medium, whereas MDC+ strains did not. Use of the medium allowed for rapid screening and isolation of mutagenized cells that had lost the ability to produce CO2 from malate. PMID:16346479

  5. Accumulation of Polyphosphate in Lactobacillus spp. and Its Involvement in Stress Resistance

    PubMed Central

    Alcántara, Cristina; Blasco, Amalia; Zúñiga, Manuel

    2014-01-01

    Polyphosphate (poly-P) is a polymer of phosphate residues synthesized and in some cases accumulated by microorganisms, where it plays crucial physiological roles such as the participation in the response to nutritional stringencies and environmental stresses. Poly-P metabolism has received little attention in Lactobacillus, a genus of lactic acid bacteria of relevance for food production and health of humans and animals. We show that among 34 strains of Lactobacillus, 18 of them accumulated intracellular poly-P granules, as revealed by specific staining and electron microscopy. Poly-P accumulation was generally dependent on the presence of elevated phosphate concentrations in the culture medium, and it correlated with the presence of polyphosphate kinase (ppk) genes in the genomes. The ppk gene from Lactobacillus displayed a genetic arrangement in which it was flanked by two genes encoding exopolyphosphatases of the Ppx-GppA family. The ppk functionality was corroborated by its disruption (LCABL_27820 gene) in Lactobacillus casei BL23 strain. The constructed ppk mutant showed a lack of intracellular poly-P granules and a drastic reduction in poly-P synthesis. Resistance to several stresses was tested in the ppk-disrupted strain, showing that it presented a diminished growth under high-salt or low-pH conditions and an increased sensitivity to oxidative stress. These results show that poly-P accumulation is a characteristic of some strains of lactobacilli and may thus play important roles in the physiology of these microorganisms. PMID:24375133

  6. Metabolic footprinting of Lactobacillus buchneri strain LA1147 during anaerobic spoilage of fermented cucumbers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Lactobacillus buchneri has recently been associated with anaerobic spoilage of fermented cucumbers due to its ability to metabolize lactic acid into acetic acid and 1,2-propanediol. However, we have limited knowledge of other chemical components in fermented cucumber that may be related to spoilage ...

  7. LIMITING GALACTOSE REQUIREMENT FOR CITRATE UTILIZATION BY LACTOBACILLUS CASEI IS ANNULLED IN CHEDDAR CHEESE EXTRACT

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Conditions required for citrate utilization by Lactobacillus casei ATCC334 were identified. Citrate is utilized by this microorganism in modified chemically defined media as an energy source, solely in the presence of limiting concentrations of other more readily metabolized carbon sources (i.e. ga...

  8. The production of glucans via glucansucrases from Lactobacillus satsumensis isolated from a fermented beverage starter culture

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Several starter cultures used in the production of fermented beverages were screened for lactic acid bacteria that produced water-insoluble polysaccharides from sucrose. The strain producing the greatest amount was identified as Lactobacillus satsumensis by its 16S RNA sequence. This strain produc...

  9. Lactobacillus plantarum MTD/1, Its Impact on Silage and In vitro Rumen Fermentation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this study was to quantify the impact of Lactobacillus plantarum MTD/1 on silage and in vitro rumen fermentation on alfalfa and corn silage. Four trials were conducted in alfalfa in second (35 and 32% DM) and third harvest (38 and 31% DM), and two in forage corn, hybrids Mycogen 797...

  10. Draft Genome Sequence of Lactobacillus plantarum WLPL04, Isolated from Human Breast Milk

    PubMed Central

    Tao, XueYing; Jiang, Meiling; Zhang, Feng; Xu, Feng

    2015-01-01

    Lactobacillus plantarum WLPL04, a novel strain, was isolated from a breast milk sample from a healthy woman and demonstrated several probiotic functions. Here, we present the draft genome sequence of this strain, which contains 3,192,587 bp, a G+C content of 44.52%, 3,158 protein-coding genes, and 53 tRNA genes. PMID:26659683

  11. Colon-specific delivery of lactobacillus rhamnosus GG using pectin hydrogel beads

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The probiotic bacteria, Lactobacillus rhamnosus GG (LGG), has shown beneficial effects on human health, and is accepted by increasing populations for the prevention and treatment of irritable bowel diseases. To increase the bioavailability and efficacy of LGG, the probiotic was encapsulated in hydro...

  12. Genome Sequence of Rapid Beer-Spoiling Isolate Lactobacillus brevis BSO 464

    PubMed Central

    Bergsveinson, Jordyn; Pittet, Vanessa; Ewen, Emily; Baecker, Nina

    2015-01-01

    The genome of brewery-isolate Lactobacillus brevis BSO 464 was sequenced and assembly produced a chromosome and eight plasmids. This bacterium tolerates dissolved CO2/pressure and can rapidly spoil packaged beer. This genome is useful for analyzing the genetics associated with beer spoilage by lactic acid bacteria. PMID:26634759

  13. Genome Sequence of a Potential Probiotic Strain, Lactobacillus fermentum HFB3, Isolated from a Human Gut

    PubMed Central

    Kumari, Madhu; Swarnkar, Mohit Kumar; Kumar, Sanjay

    2015-01-01

    A draft genome sequence of 2.04 Mb is reported for Lactobacillus fermentum HFB3, which is a lactic acid bacterium with probiotic properties. The gene-coding clusters also predicted the presence of genes responsible for probiotic characteristics. PMID:26543124

  14. ISOLATION OF A LACTOBACILLUS SALIVARIUS: ITS BACTERIOCIN IS INHIBITORY TO CAMPYLOBACTER JEJUNI IN CHICKENS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We evaluated anti-Campylobacter jejuni activity among >1,200 isolates of lactic acid bacteria. One, Lactobacillus salivarius (NRRL B-30514), was selected for further study. The cell-free, ammonium sulfate precipitate from the broth culture was termed the crude antimicrobial preparation (CAP). Ten...

  15. Effect of intestinal colonisation by two Lactobacillus strains on the immune response of gnotobiotic mice.

    PubMed

    Steinberg, R S; Lima, M; Gomes de Oliveira, N L; Miyoshi, A; Nicoli, J R; Neumann, E; Nunes, A C

    2014-12-01

    The effect of intestinal colonisation on the immune system was investigated in germ-free mice monoassociated with Lactobacillus strains isolated from calf faeces. Single doses of Lactobacillus acidophilus L36 or Lactobacillus salivarius L38 were administered to germ-free mice by intragastric gavage. Ten days later, the mice were euthanised. Gene expression levels of interleukin 5 (IL-5), IL-6, IL-10, IL-12b, IL-17a, gamma interferon (IFN-?), transforming growth factor beta 1 (TGF-?1), and tumour necrosis factor alpha (TNF-?) were quantified in segments of the small and large intestines by real time quantitative polymerase chain reaction. All the mice were colonised rapidly after Lactobacillus administration with intestinal counts ranging from 6.53 to 8.26 log cfu/g. L. acidophilus L36 administration increased the expression of cytokines involved with the Th2 (IL-5, IL-6 and TGF-?1) and Th17 (IL-17a, TNF-? and IL-6) inflammatory response, whereas L. salivarius L38 appeared to stimulate a pattern of less diversified cytokines in the intestine. Intragastric gavage of L. acidophilus L36 and L. salivarius L38 induced similar levels of colonisation in the digestive tracts of germ-free mice but stimulated different immune responses in the intestinal mucosa. The different immunomodulation patterns might facilitate the potential use of these lactobacilli as probiotics to treat distinct pathological conditions, for example protection against Citrobacter rodentium infection by stimulating IL-17 production. PMID:24939801

  16. Proteomic Analyses of Ethanol Tolerance in Lactobacillus buchneri NRRL B-30929

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Lactobacillus buchneri NRRL B-30929 strain, isolated from a fuel ethanol production facility, exhibits high tolerance to environmental ethanol concentrations. In this study, the ethanol tolerance trait was elucidated at the molecular level by using proteomics comparison and analyses. Cellular p...

  17. Characterization of Lactobacillus fermentum PL9988 Isolated from Healthy Elderly Korean in a Longevity Village.

    PubMed

    Park, Jong-Su; Shin, Eunju; Hong, Hyunjin; Shin, Hyun-Jung; Cho, Young-Hoon; Ahn, Ki-Hyun; Paek, Kyungsoo; Lee, Yeonhee

    2015-09-28

    In this work, we wanted to develop a probiotic from famous longevity villages in Korea. We visited eight longevity villages in Korea to collect fecal samples from healthy adults who were aged above 80 years and had regular bowel movements, and isolated lactic-acid-producing bacteria from the samples. Isolated colonies that appeared on MRS agar containing bromophenol blue were identified by means of 16S rRNA sequencing, and 102 of the isolates were identified as lactic-acid-producing bacteria (18 species). Lactobacillus fermentum was the most frequently found species. Eight isolates were selected on the basis of their ability to inhibit the growth of six intestinal pathogens (Escherichia coli O157:H7, Salmonella enterica subsp. enterica Typhimurium, Salmonella enterica subsp. enterica Enteritidis, Enterococcus faecalis, Staphylococcus aureus, and Listeria monocytogenes) and their susceptibility to 15 antimicrobial agents. Among these eight isolates, four Lactobacillus fermentum isolates were found not to produce any harmful enzymes or metabolites. Among them, Lactobacillus fermentum isolate no. 24 showed the strongest binding to intestinal epithelial cells, the highest immune-enhancing activity, anti-inflammation activity, and anti-oxidation activity as well as the highest survival rates in the presence of artificial gastric juice and bile solution. This isolate, designated Lactobacillus fermentum PL9988, has all the characteristics for a good probiotic. PMID:26095384

  18. Preparation of a Lactobacillus plantarum starter culture for cucumber fermentations that can meet kosher guidelines

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A method is described for growth of a Lactobacillus plantarum starter culture in jars of commercially available pasteurized fresh-pack kosher dill cucumbers so that jars can be used to inoculate commercial scale cucumber fermentation tanks. A procedure is also described to transfer lactic acid bacte...

  19. Coexpression of pyruvate decarboxylase and alcohol dehydrogenase genes in Lactobacillus brevis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Lactobacillus brevis ATCC 367 is able to metabolize xylose into lactate and acetate but not ethanol. In an attempt to transform L. brevis into an ethanologen that uses xylose, a Gram-positive gene for pyruvate decarboxylase (PDC) was introduced. This enzyme catalyzes the decarboxylation of pyruvat...

  20. Inactivation of Lactobacillus plantarum in apple cider using radio frequency electric fields

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Radio frequency electric fields (RFEF) processing is effective at inactivating Gram negative bacteria in fruit juices at moderately low temperatures, but has yet to be shown to be effective at reducing Gram positive bacteria. Lactobacillus plantarum ATCC 49445, a Gram positive bacterium, was inocula...

  1. Genome Sequence of Lactobacillus gastricus PS3, a Strain Isolated from Human Milk.

    PubMed

    Martín, Virginia; Cárdenas, Nivia; Jiménez, Esther; Maldonado, Antonio; Rodríguez, Juan Miguel; Fernández, Leonides

    2013-01-01

    Lactobacillus gastricus is a mostly unknown lactobacilli species associated with mucosal surfaces. We present the draft annotated genome sequence of L. gastricus strain PS3, isolated from a human milk sample, to provide new insights into its biology and to characterize those genes related to advantageous technological and beneficial properties. PMID:23846278

  2. Carbohydrate utilization and detection of a nucleotide hydrolase in Lactobacillus buchneri

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Lactobacillus buchneri strains NRRL 1837, DSM 5987, and NRRL B-30929 were examined for capacity to metabolize various carbohydrates via growth and fermentation analyses. Carbon sources used for this study included D-melibiose, inosine, uridine, D-melezitose, maltotriose, N-acetyl-D-glucosamine, suc...

  3. Physiological Characteristics and Anti-obesity Effect of Lactobacillus plantarum Q180 Isolated from Feces

    PubMed Central

    Park, Sun-Young; Cho, Seong-A; Kim, Sae-Hun; Lim, Sang-Dong

    2014-01-01

    Obesity is strongly associated with several metabolic and chronic diseases and has become a major public health problem of worldwide concern. This study aimed to investigate the physiological characteristics and anti-obesity effects of Lactobacillus plantarum Q180. Lactobacillus plantarum Q180 was isolated from the faces of healthy adults and found to have a lipase inhibitory activity of 83.61±2.32% and inhibited adipocyte differentiation of 3T3-L1 cells (14.63±1.37%) at a concentration of 100 ?g/mL. The strain was investigated for its physiological characteristics. The optimum growth temperature of L. plantarum Q180 was 37?. Lactobacillus plantarum Q180 showed higher sensitivity to novobiocin in a comparison of fifteen different antibiotics and showed the highest resistance to rifampicin, polymyxin B and vancomycin. The strain showed higher ?-galactosidase and N-acetyl-?-glucosaminidase activities. It also did not produce carcinogenic enzymes such as ?-glucuronidase. The survival rate of L. plantarum Q180 in MRS broth containing 0.3% bile was 97.8%. Moreover, the strain showed a 97.2% survival rate after incubation for 3 h in pH 2.0. Lactobacillus plantarum Q180 was displayed resistance to Escherichia coli, Salmonella Typhimurium and Staphylococcus aureus with rates of 55.6%, 38.0% and 47.6%, respectively. These results demonstrate that L. plantarum Q180 has potential as a probiotic with anti-obesity effects.

  4. EFFECT OF SALT NUTRIENTS ON MANNITOL PRODUCTION BY LACTOBACILLUS INTERMEDIUS NRRL B-3693

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effects of four salt nutrients (ammonium citrate, sodium phosphate, magnesium sulfate, and manganese sulfate) on the production of mannitol by Lactobacillus intermedius NRRL B-3693 in a simplified medium containing 300 g fructose, 5 g soy peptone, and 50 g corn steep liquor per L in pH-controlle...

  5. Draft Genome Sequence of Lactobacillus hominis Strain CRBIP 24.179T, Isolated from Human Intestine

    PubMed Central

    Cousin, Sylvie; Creno, Sophie; Ma, Laurence; Clermont, Dominique; Loux, Valentin; Bizet, Chantal

    2013-01-01

    We report the draft genome sequence of the strain Lactobacillus hominis CRBIP 24.179T, isolated from a human clinical sample. The total length of the 28 contigs is about 1.9 Mb, with a G+C content of 37% and 1,983 coding sequences. PMID:23969062

  6. Draft Genome Sequence of Lactobacillus gigeriorum CRBIP 24.85T, Isolated from a Chicken Crop

    PubMed Central

    Ma, Laurence; Creno, Sophie; Clermont, Dominique; Loux, Valentin; Bizet, Chantal; Bouchier, Christiane

    2012-01-01

    We report the draft genome of the strain Lactobacillus gigeriorum CRBIP 24.85T, isolated from a chicken crop. The total length of the 60 scaffolds is about 1.9 Mb, with a GC content of 38% and 2,062 protein-coding sequences (CDS). PMID:23045490

  7. Draft Genome Sequence of Lactobacillus pasteurii CRBIP 24.76T

    PubMed Central

    Cousin, Sylvie; Clermont, Dominique; Creno, Sophie; Ma, Laurence; Loux, Valentin; Bizet, Chantal

    2013-01-01

    We report the draft genome sequence of the type strain Lactobacillus pasteurii CRBIP 24.76, which is closely related to L. gigeriorum CRBIP 24.85T, isolated from a chicken crop. The total length of the 29 contigs is about 1.9 Mb, with a G+C content of 40% and 1,946 coding sequences. PMID:23969061

  8. Draft Genome Sequence of Lactobacillus johnsonii Strain 16, Isolated from Mice

    PubMed Central

    Buhnik-Rosenblau, Keren; Danin-Poleg, Yael; Elgavish, Sharona

    2015-01-01

    Here, we report the genome sequence of Lactobacillus johnsonii, a member of the gut lactobacilli. This draft genome of L. johnsonii strain 16 isolated from C57BL/6J mice enables the identification of bacterial genes responsible for host-specific gut persistence. PMID:26450724

  9. Genome Sequence of Rough and Smooth Variants of Pleomorphic Strain Lactobacillus farciminis CNCM-I-3699

    PubMed Central

    Tareb, R.; Bernardeau, M.

    2015-01-01

    The probiotic Lactobacillus farciminis CNCM-I-3699 is a pleomorphic strain exhibiting smooth and rough variants. We report their complete genomes consisting of a chromosome of 2, 4 Mb and a plasmid of 6,417 bp. The smooth variant differs by the presence of an additional plasmid of 35,418 bp. PMID:26383668

  10. Draft Genome Sequence of a Probiotic Strain, Lactobacillus fermentum UCO-979C

    PubMed Central

    Villena, Julio; Gonzalez, Carlos; Albarracin, Leonardo; Barros, Javier

    2015-01-01

    This report describes a draft genome sequence of Lactobacillus fermentum strain UCO-979C. The reads generated by a Ion Torrent PGM were assembled into contigs, with a total size of 2.01 Mb. The data were annotated using the NCBI GenBank and RAST servers. Specific features of the genome are highlighted. PMID:26659681

  11. Genome Sequence of Rapid Beer-Spoiling Isolate Lactobacillus brevis BSO 464.

    PubMed

    Bergsveinson, Jordyn; Pittet, Vanessa; Ewen, Emily; Baecker, Nina; Ziola, Barry

    2015-01-01

    The genome of brewery-isolate Lactobacillus brevis BSO 464 was sequenced and assembly produced a chromosome and eight plasmids. This bacterium tolerates dissolved CO2/pressure and can rapidly spoil packaged beer. This genome is useful for analyzing the genetics associated with beer spoilage by lactic acid bacteria. PMID:26634759

  12. pH-induced Structural Changes Regulate Histidine Decarboxylase Activity in Lactobacillus 30a

    E-print Network

    pH-induced Structural Changes Regulate Histidine Decarboxylase Activity in Lactobacillus 30a acids, and to optimize cell growth. The HDC trimer is active at low pH and inactive at neutral to alkaline pH. We have solved the X-ray structure of HDC at pH 8 and revealed the novel mechanism of pH

  13. Complete genome sequence of Lactobacillus acetotolerans RIB 9124 (NBRC 13120) isolated from putrefied (hiochi) Japanese sake.

    PubMed

    Toh, Hidehiro; Morita, Hidetoshi; Tsuji, Hiroyuki; Iwashita, Kazuhiro; Goto, Nami; Nakayama, Jiro; Sekine, Mitsuo; Kato, Yumiko; Suzuki, Ken-Ichiro; Fujita, Nobuyuki

    2015-11-20

    Lactobacillus acetotolerans RIB 9124 (NBRC 13120) was isolated from putrefied (hiochi) Japanese sake. Here we report the complete genome sequence of this organism. This paper is the first report demonstrating the fully sequenced and completely annotated genome of a L. acetotolerans strain. PMID:26376472

  14. Conversion of Biomass Hydrolysates and Other Substrates to Ethanol and Other Chemicals by Lactobacillus buchneri

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A Lactobacillus buchneri strain NRRL B-30929 can convert xylose and glucose into ethanol and chemicals. In this paper, L. buchneri NRRL B-30929 was initially compared with the type strains L. buchneri NRRL 1837 and DSM 5987 for growth and fermentation using single substrate derived from plant mater...

  15. Milk production response to feeding alfalfa silage inoculated with Lactobacillus plantarum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In mini-silo trials, silages treated with a Lactobacillus plantarum silage inoculant (Ecosyl, Yorkshire, UK) had increased in vitro rumen microbial biomass production compared to untreated. Our objective was to determine if alfalfa silage treated with this inoculant could produce a milk production r...

  16. Use of a lactobacillus-based probiotic culture to reduce Salmonella in day of hatch broilers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A commercially available Lactobacillus probiotic (FM-B11™ Ivesco LLC) (B11) significantly reduced Salmonella recovery from day-of-hatch chicks in several studies. For all experiments, day-of-hatch male broiler chicks (n=40 per pen) were challenged with approximately 10**4 cfu per chick of Salmonell...

  17. Draft Genome Sequence of a Probiotic Strain, Lactobacillus fermentum UCO-979C.

    PubMed

    Karlyshev, Andrey V; Villena, Julio; Gonzalez, Carlos; Albarracin, Leonardo; Barros, Javier; Garcia, Apolinaria

    2015-01-01

    This report describes a draft genome sequence of Lactobacillus fermentum strain UCO-979C. The reads generated by a Ion Torrent PGM were assembled into contigs, with a total size of 2.01 Mb. The data were annotated using the NCBI GenBank and RAST servers. Specific features of the genome are highlighted. PMID:26659681

  18. Safety and persistence of orally administered human Lactobacillus sp. strains in healthy adults.

    PubMed

    Hütt, P; Kõll, P; Stsepetova, J; Alvarez, B; Mändar, R; Krogh-Andersen, K; Marcotte, H; Hammarström, L; Mikelsaar, M

    2011-03-01

    The aim of the study was to evaluate the safety and persistence of selected Lactobacillus strains in the gastrointestinal tract (GIT) of healthy adult volunteers after oral consumption of high doses of lactobacilli to identify potential candidates for probiotic and biotechnological applications. In the first phase of the study, nine individuals consumed capsules containing Lactobacillus gasseri 177 and E16B7, Lactobacillus acidophilus 821-3, Lactobacillus paracasei 317 and Lactobacillus fermentum 338-1-1 (each daily dose 1×1010 cfu) for 5 consecutive days. Data on gut health, blood parameters, and liver and kidney function were collected. The persistence of Lactobacillus strains was assessed by culturing combined with arbitrarily primed polymerase chain reaction (AP-PCR) and PCR-denaturing gradient gel electrophoresis (PCR-DGGE) on days 0, 5, 8, 10 and 20 from faecal samples. All strains survived gastrointestinal passage and were detected on the 5th day. L. acidophilus 821-3 was detected in four volunteers on the 8th day (4.3 to 7.0 log10 cfu/g) and in two on the 10th day (8.3 and 3.9 log10 cfu/g, respectively). In the second phase of the study, five additional volunteers consumed L. acidophilus 821-3 (daily 1×1010 cfu) for 5 consecutive days. The strain was subsequently detected in faeces of all individuals using real-time PCR on the 10th day (range 4.6-6.7; median 6.0 log10 cell/g) in both phases of the study for at least 5 days after discontinuation of consumption. The administration of high doses of different Lactobacillus strains did not result in any severe adverse effects in GIT and/or abnormal values of blood indices. Thus, the strain L. acidophilus 821-3 is a promising candidate for probiotic and biotechnological applications. Further studies will be performed to confirm the strain persistence and safety in a larger number of individuals. PMID:21831792

  19. Antibacterial properties of composite resins incorporating silver and zinc oxide nanoparticles on Streptococcus mutans and Lactobacillus

    PubMed Central

    Kasraei, Shahin; Sami, Lida; Hendi, Sareh; AliKhani, Mohammad-Yousef; Rezaei-Soufi, Loghman

    2014-01-01

    Objectives Recurrent caries was partly ascribed to lack of antibacterial properties in composite resin. Silver and zinc nanoparticles are considered to be broad-spectrum antibacterial agents. The aim of the present study was to evaluate the antibacterial properties of composite resins containing 1% silver and zinc-oxide nanoparticles on Streptococcus mutans and Lactobacillus. Materials and Methods Ninety discoid tablets containing 0%, 1% nano-silver and 1% nano zinc-oxide particles were prepared from flowable composite resin (n = 30). The antibacterial properties of composite resin discs were evaluated by direct contact test. Diluted solutions of Streptococcus mutans (PTCC 1683) and Lactobacillus (PTCC 1643) were prepared. 0.01 mL of each bacterial species was separately placed on the discs. The discs were transferred to liquid culture media and were incubated at 37? for 8 hr. 0.01 mL of each solution was cultured on blood agar and the colonies were counted. Data was analyzed with Kruskall-Wallis and Mann-Whitney U tests. Results Composites containing nano zinc-oxide particles or silver nanoparticles exhibited higher antibacterial activity against Streptococcus mutans and Lactobacillus compared to the control group (p < 0.05). The effect of zinc-oxide on Streptococcus mutans was significantly higher than that of silver (p < 0.05). There were no significant differences in the antibacterial activity against Lactobacillus between composites containing silver nanoparticles and those containing zinc-oxide nanoparticles. Conclusions Composite resins containing silver or zinc-oxide nanoparticles exhibited antibacterial activity against Streptococcus mutans and Lactobacillus. PMID:24790923

  20. Zymogram and Preliminary Characterization of Lactobacillus helveticus Autolysins

    PubMed Central

    Valence, F.; Lortal, S.

    1995-01-01

    The autolysins of Lactobacillus helveticus ISLC5 were detected and partially characterized by renaturing sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis with substrate-containing gels (zymogram). By using lyophilized Micrococcus luteus cells or heated whole cells of L. helveticus ISLC5 (0.2% [wt/vol]) as a substrate, several lytic activities were detected in the whole-cell SDS extract of strain ISLC5 (i) one activity at 42.4 kDa, which was named autolysin A, and (ii) six other activities having very similar molecular weights (29.1, 29.6, 30, 30.8, 31.7, and 32.8 kDa), which were named autolysins B (B1 through B6, respectively). As regards the temporal distribution of the enzymes, autolysins A and B were detected in the cells harvested from the beginning of the exponential growth phase. Autolysin A appeared to be associated only with viable cells, whereas the autolysins B remained associated with the cell envelope several days after the complete loss of culture viability. When SDS-treated walls of L. helveticus ISLC5 were used as a substrate, a supplementary lytic activity appeared at 37.5 kDa; it was considered a peptidoglycan hydrolase, since it was not able to induce lysis of whole-cell substrate. The autolysins of 30 other strains of L. helveticus from various geographical origins were also analyzed by zymogram; all the activity profiles obtained were similar to that of strain ISLC5 in terms of the number of lytic bands and their apparent molecular weights. Only the relative intensities of the lytic bands corresponding to autolysins A and B were variable depending on the strains. This observation suggested that autolysins are highly conserved enzymes. A concentrated crude lysate of the virulent bacteriophage 832-B1 infecting L. helveticus was also analyzed by zymogram; one lytic activity with an apparent molecular weight of 31.7 kDa, very close to the weights of the autolysins B, was observed. Finally, the autolysins of L. helveticus ISLC5 were successfully extracted from whole cells by using a 1 M lithium chloride solution; they were partially purified by precipitation, selective resolubilization, and gel filtration chromatography, which led to a 20-fold increase in specific activity. PMID:16535125

  1. PRODUCTION OF MANNITOL BY LACTOBACILLUS INTERMEDIUS NRRL B-3693 IN FED-BATCH AND CONTINUOUS CELL-RECYCLE FERMENTATIONS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Improved fermentation processes were developed for the production of mannitol by a heterofermentative lactic acid bacterium (Lactobacillus intermedius NRRL B-3693). A fed-batch fermentation protocol overcame limitations caused by high substrate concentrations. The process was developed using prima...

  2. Enzymatic fractionation of the antimicrobial peptides casocidin and isracidin by Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The cumulative effect of peptidase and protease activities associated with cells of Streptococcus thermophilus (ST) and Lactobacillus delbrueckii subsp. bulgaricus (LB) was evaluated on the milk-protein based antimicrobial peptides casocidin and isracidin. Reaction mixtures of casocidin or isracidin...

  3. Characterization of a bovine isolate Lactobacillus mucosae DPC 6426 which produces an exopolysaccharide composed predominantly of mannose residues

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aim: Identification of exopolysaccharide (EPS)-producing lactobacilli as EPS production is potentially a very important trait among probiotic lactobacilli from technological and health promoting perspectives. Methods and Results: Characterization of EPS-producing Lactobacillus mucosae DPC 6426 in de...

  4. PEDIOCIN PRODUCTION IN MILK BY PEDIOCOCCUS ACIDILACTICI IN CO-CULTURE WITH STREPTOCOCCUS THERMOPHILUS AND LACTOBACILLUS DELBRUECKII SUBSP. BULGARICUS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The production of pediocin in milk by Pediococcus acidilactici was evaluated in co-culture with the dairy fermentation cultures Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, and Lactococcus lactis subsp. lactis. The cultures were tested singly or in different combinations...

  5. In vitro probiotic characterization of Lactobacillus strains from fermented radish and their anti-adherence activity against enteric pathogens.

    PubMed

    Damodharan, Karthiyaini; Palaniyandi, Sasikumar Arunachalam; Yang, Seung Hwan; Suh, Joo-Won

    2015-11-01

    In this study, we evaluated the probiotic properties of Lactobacillus plantarum, Lactobacillus pentosus, and Lactobacillus fermentum strains isolated from fermented radish. All the strains survived the simulated oro-gastrointestinal transit condition and showed significantly higher adherence to Caco-2 cells compared with the probiotic strain Lactobacillus rhamnosus GG. The strains showed broad-spectrum antimicrobial activity, autoaggregation, and coaggregation capacity with pathogens. Furthermore, the Lactobacillus strains inhibited the adherence of Yersinia enterocolitica subsp. enterocolitica, Shigella boydii, and Salmonella choleraesuis to the Caco-2 cell line. The strains possessed bile salt hydrolase activity and their cholesterol-lowering activity in vitro was above 50% in the presence of bile. Strains of L. plantarum and L. pentosus possessed the plantaricin-encoding plnEF gene. In addition, the Lactobacillus strains maintained about 80% cell viability after freeze-drying in the presence of a combination of 5% skim milk and 5% maltodextrin as cryoprotectant, and 70% recovery of cell viability was observed in the absence of any cryoprotectant. PMID:26382558

  6. Lactobacillus gasseri requires peptides, not proteins or free amino acids, for growth in milk.

    PubMed

    Arakawa, K; Matsunaga, K; Takihiro, S; Moritoki, A; Ryuto, S; Kawai, Y; Masuda, T; Miyamoto, T

    2015-03-01

    Lactobacillus gasseri is a widespread commensal lactic acid bacterium inhabiting human mucosal niches and has many beneficial effects as a probiotic. However, L. gasseri is difficult to grow in milk, which hurts usability for the food industry. It had been previously reported that supplementation with yeast extract or proteose peptone, including peptides, enables L. gasseri to grow well in milk. In this study, our objective was to confirm peptide requirement of L. gasseri and evaluate efficacy of peptide release by enzymatic proteolysis on growth of L. gassei in milk. Three strains of L. gasseri did not grow well in modified DeMan, Rogosa, Sharpe broth without any nitrogen sources (MRS-N), but addition of a casein-derived peptide mixture, tryptone, promoted growth. In contrast, little effect was observed after adding casein or a casein-derived amino acid mixture, casamino acids. These results indicate that L. gasseri requires peptides, not proteins or free amino acids, among milk-derived nitrogen sources for growth. Lactobacillus gasseri JCM 1131T hardly had growth capacity in 6 kinds of milk-based media: bovine milk, human milk, skim milk, cheese whey, modified MRS-N (MRSL-N) supplemented with acid whey, and MRSL-N supplemented with casein. Moreover, treatment with digestive proteases, particularly pepsin, to release peptides made it grow well in each milk-based medium. The pepsin treatment was the most effective for growth of strain JCM 1131T in skim milk among the tested food-grade proteases such as trypsin, ?-chymotrypsin, calf rennet, ficin, bromelain, and papain. As well as strain JCM 1131T, pepsinolysis of milk improved growth of other L. gasseri strains and some strains of enteric lactobacilli such as Lactobacillus crispatus, Lactobacillus gallinarum, Lactobacillus johnsonii, and Lactobacillus reuteri. These results suggest that some relatives of L. gasseri also use peptides as desirable nitrogen sources, and that milk may be a good supplier of nutritious peptides to enteric lactobacilli including L. gasseri after peptic digestion in the gastrointestinal tract. This is the first report showing peptide requirement of L. gasseri and efficacy of pepsinolysis on the growth of L. gasseri and its relatives in milk. This study would contribute to increasing usability of L. gasseri and its relatives as probiotics in dairy foods. PMID:25529420

  7. Lactobacillus rhamnosus L34 and Lactobacillus casei L39 suppress Clostridium difficile-induced IL-8 production by colonic epithelial cells

    PubMed Central

    2014-01-01

    Background Clostridium difficile is the main cause of hospital-acquired diarrhea and colitis known as C. difficile-associated disease (CDAD).With increased severity and failure of treatment in CDAD, new approaches for prevention and treatment, such as the use of probiotics, are needed. Since the pathogenesis of CDAD involves an inflammatory response with a massive influx of neutrophils recruited by interleukin (IL)-8, this study aimed to investigate the probiotic effects of Lactobacillus spp. on the suppression of IL-8 production in response to C. difficile infection. Results We screened Lactobacillus conditioned media from 34 infant fecal isolates for the ability to suppress C. difficile-induced IL-8 production from HT-29 cells. Factors produced by two vancomycin-resistant lactobacilli, L. rhamnosus L34 (LR-L34) and L.casei L39 (LC-L39), suppressed the secretion and transcription of IL-8 without inhibiting C. difficile viability or toxin production. Conditioned media from LR-L34 suppressed the activation of phospho-NF-?B with no effect on phospho-c-Jun. However, LC-L39 conditioned media suppressed the activation of both phospho-NF-?B and phospho-c-Jun. Conditioned media from LR-L34 and LC-L39 also decreased the production of C. difficile-induced GM-CSF in HT-29 cells. Immunomodulatory factors present in the conditioned media of both LR-L34 and LC-L39 are heat-stable up to 100°C and?>?100 kDa in size. Conclusions Our results suggest that L. rhamnosus L34 and L. casei L39 each produce factors capable of modulating inflammation stimulated by C. difficile. These vancomycin-resistant Lactobacillus strains are potential probiotics for treating or preventing CDAD. PMID:24989059

  8. Quantitative transcript analysis of the inducible expression system pSIP: comparison of the overexpression of Lactobacillus spp. ?-galactosidases in Lactobacillus plantarum

    PubMed Central

    2011-01-01

    Background Two sets of overlapping genes, lacLMReu and lacLMAci, encoding heterodimeric ?-galactosidases from Lactobacillus reuteri and Lactobacillus acidophilus, respectively, have previously been cloned and expressed using the pSIP vector system and Lactobacillus plantarum WCSF1 as host. Despite the high similarity between these lacLM genes and the use of identical cloning and expression strategies, strains harboring lacLMReu produced about twenty-fold more ?-galactosidase than strains containing lacLMAci. Results In this study, the plasmid copy numbers (PCN) of expression vectors pEH9R (lacLMReu) and pEH9A (lacLMAci) as well as the transcription levels of both lacLM genes were compared using quantitative PCR methods. Analyses of parallel fermentations of L. plantarum harboring either pEH9R or pEH9A showed that the expression plasmids were present in similar copy numbers. However, transcript levels of lacLM from L. reuteri (pEH9R) were up to 18 times higher than those of lacLM from L. acidophilus (pEH9A). As a control, it was shown that the expression levels of regulatory genes involved in pheromone-induced promoter activation were similar in both strains. Conclusion The use of identical expression strategies for highly similar genes led to very different mRNA levels. The data indicate that this difference is primarily caused by translational effects that are likely to affect both mRNA synthesis rates and mRNA stability. These translational effects thus seem to be a dominant determinant for the success of gene expression efforts in lactobacilli. PMID:21696579

  9. Use of green fluorescent protein to monitor Lactobacillus plantarum in the gastrointestinal tract of goats

    PubMed Central

    Han, Xufeng; Wang, Lei; Li, Wei; Li, Bibo; Yang, Yuxin; Yan, Hailong; Qu, Lei; Chen, Yulin

    2015-01-01

    The experiment aimed to specifically monitor the passage of lactobacilli in vivo after oral administration. The green fluorescent protein (GFP) gene was cloned downstream from the constitutive p32 promoter from L. lactis subsp. cremoris Wg2. The recombinant expression vector, pLEM415-gfp-p32, was electroporated into Lactobacillus plantarum (L. plantarum) isolated from goat. Green fluorescent protein (GFP) was successfully expressed in L. plantarum. After 2 h post-administration, transformed Lactobacillus could be detectable in all luminal contents. In the rumen, bacteria concentration initially decreased, reached the minimum at 42 h post-oral administration and then increased. However, this concentration decreased constantly in the duodenum. This result indicated that L. plantarum could colonize in the rumen but not in the duodenum. PMID:26413069

  10. Use of green fluorescent protein to monitor Lactobacillus plantarum in the gastrointestinal tract of goats.

    PubMed

    Han, Xufeng; Wang, Lei; Li, Wei; Li, Bibo; Yang, Yuxin; Yan, Hailong; Qu, Lei; Chen, Yulin

    2015-01-01

    The experiment aimed to specifically monitor the passage of lactobacilli in vivo after oral administration. The green fluorescent protein (GFP) gene was cloned downstream from the constitutive p32 promoter from L. lactis subsp. cremoris Wg2. The recombinant expression vector, pLEM415-gfp-p32, was electroporated into Lactobacillus plantarum (L. plantarum) isolated from goat. Green fluorescent protein (GFP) was successfully expressed in L. plantarum. After 2 h post-administration, transformed Lactobacillus could be detectable in all luminal contents. In the rumen, bacteria concentration initially decreased, reached the minimum at 42 h post-oral administration and then increased. However, this concentration decreased constantly in the duodenum. This result indicated that L. plantarum could colonize in the rumen but not in the duodenum. PMID:26413069

  11. Lactobacillus priming of the respiratory tract: Heterologous immunity and protection against lethal pneumovirus infection.

    PubMed

    Garcia-Crespo, Katia E; Chan, Calvin C; Gabryszewski, Stanislaw J; Percopo, Caroline M; Rigaux, Peter; Dyer, Kimberly D; Domachowske, Joseph B; Rosenberg, Helene F

    2013-03-01

    We showed previously that wild-type mice primed via intranasal inoculation with live or heat-inactivated Lactobacillus species were fully (100%) protected against the lethal sequelae of infection with the virulent pathogen, pneumonia virus of mice (PVM), a response that is associated with diminished expression of proinflammatory cytokines and diminished virus recovery. We show here that 40% of the mice primed with live Lactobacillus survived when PVM challenge was delayed for 5months. This robust and sustained resistance to PVM infection resulting from prior interaction with an otherwise unrelated microbe is a profound example of heterologous immunity. We undertook the present study in order to understand the nature and unique features of this response. We found that intranasal inoculation with L. reuteri elicited rapid, transient neutrophil recruitment in association with proinflammatory mediators (CXCL1, CCL3, CCL2, CXCL10, TNF-alpha and IL-17A) but not Th1 cytokines. IFN? does not contribute to survival promoted by Lactobacillus-priming. Live L. reuteri detected in lung tissue underwent rapid clearance, and was undetectable at 24h after inoculation. In contrast, L. reuteri peptidoglycan (PGN) and L. reuteri genomic DNA (gDNA) were detected at 24 and 48h after inoculation, respectively. In contrast to live bacteria, intranasal inoculation with isolated L. reuteri gDNA elicited no neutrophil recruitment, had minimal impact on virus recovery and virus-associated production of CCL3, and provided no protection against the negative sequelae of virus infection. Isolated PGN elicited neutrophil recruitment and proinflammatory cytokines but did not promote sustained survival in response to subsequent PVM infection. Overall, further evaluation of the responses leading to Lactobacillus-mediated heterologous immunity may provide insight into novel antiviral preventive modalities. PMID:23274789

  12. Draft Genome Sequence of Lactobacillus plantarum Lp90 Isolated from Wine

    PubMed Central

    Lamontanara, Antonella; Caggianiello, Graziano; Orrù, Luigi; Capozzi, Vittorio; Michelotti, Vania; Bayjanov, Jumamurat R.; Renckens, Bernadet; van Hijum, Sacha A. F. T.; Cattivelli, Luigi

    2015-01-01

    Here, we describe the draft genome sequence and annotation of Lactobacillus plantarum strain Lp90, the first sequenced genome of a L. plantarum strain isolated from wine. This strain has a noticeable ropy phenotype and showed potential probiotic properties. The genome consists of 3,324,076 bp (33 contigs) and contains 3,155 protein coding genes, 34 pseudogenes, and 84 RNA genes. PMID:25767234

  13. Complete genome sequence of Lactobacillus helveticus MB2-1, a probiotic bacterium producing exopolysaccharides.

    PubMed

    Li, Wei; Xia, Xiudong; Chen, Xiaohong; Rui, Xin; Jiang, Mei; Zhang, Qiuqin; Zhou, Jianzhong; Dong, Mingsheng

    2015-09-10

    Lactobacillus helveticus MB2-1 is a probiotic bacterium producing exopolysaccharides (EPS), which was isolated from traditional Sayram ropy fermented milk in southern Xinjiang, China. The genome consists of a circular 2,084,058bp chromosome with no plasmid. The genome sequence indicated that this strain includes a 15.20kb gene cluster involved in EPS biosynthesis. Genome sequencing information has provided the basis for understanding the potential molecular mechanism behind the EPS production. PMID:26065338

  14. Probiotic features of Lactobacillus strains isolated from Ragusano and Pecorino Siciliano cheeses.

    PubMed

    Caggia, C; De Angelis, M; Pitino, I; Pino, A; Randazzo, C L

    2015-09-01

    In the present study 177 Lactobacillus spp. strains, isolated from Ragusano and Pecorino Siciliano cheeses, were in vitro screened for probiotic traits, and their characteristics were compared to those of Lactobacillus rhamnosus GG, commercial strain. Based on acidic and bile salt resistance, thirteen Lactobacillus strains were selected. The multiplex-PCR application revealed that nine strains belonged to L. rhamnosus species and four to Lactobacillus paracasei species. All selected strains were further investigated for transit tolerance in simulated upper gastrointestinal tract (GI), for adhesion capacity to human intestinal cell lines, for hydrophobicity, for co-aggregation and auto-aggregation and for antimicrobial activities. Moreover, antibiotic resistance, hemolytic and bile salt hydrolase activities were investigated for safety assessment. Viable counts after simulated gastric and duodenal transit revealed that overall the selected lactobacilli tolerated better pancreatic juice and bile salts than acidic juice. In particular, three L. rhamnosus strains (FS10, FS2, and PS11) and one L. paracasei strain (PM8) increased their cell density after the simulated GI transit. The same strains showed also high percentage of auto-aggregation and co-aggregation with Escherichia coli. All strains were effective against both Staphylococcus aureus and E. coli and variability was achieved versus Listeria monocytogenes and Enterococcus faecalis used as pathogenic indicator strains. Different behavior was revealed by strains for adhesion ability and hydrophobicity, which are not always linked each other and are strongly strain-dependent. From the safety point of view, no isolate showed hemolytic and bile salt hydrolase activities, except one, and most of the strains were sensitive to a broad range of clinical antibiotics. This work showed that the L. rhamnosus FS10 and the L. paracasei PM8 are good promising probiotic candidates for further in vivo investigations. PMID:25998823

  15. Assessment of Characteristics and Functional Properties of Lactobacillus Species Isolated from Kimchi for Dairy Use

    PubMed Central

    Baick, Seung-Chun

    2015-01-01

    The objective of this study was to identify lactic acid bacteria (LAB) isolated from kimchi and to evaluate its characteristics and functional properties for application in fermented dairy products as a probiotic or commercial starter culture. Eight stains isolated from kimchi were selected through an investigation of phenotypic characteristics. Two strains (DK211 and DK303) were identified as Lactobacillus plantarum, another two (DK207 and DK215) as Lactobacillus paracasei, and one (DK301) as Lactobacillus sakei. The remaining three strains were identified as species of Weissella. All selected Lactobacillus strains had acid and bile tolerance, even though there was wide variation in the ability of each strain. DK303 showed a remarkably higher proteolytic activity. There were no significant differences in ?-galactosidase activity among the tested strains, except that DK301 showed no activity. Auto-aggregation varied between 82.1 and 90.0%, and hydrophobicity values ranged from 0.5 to 51.6%.The strongest auto-aggregation and hydrophobicity were observed in DK211. All selected strains showed better 1,1-diphenyl-2-picrylhydrzyl (DPPH) scavenging activity than commercial strains. DK211, DK215, DK301, and DK303 had effective inhibitory activity against all pathogens tested except E. coli. When selected strains were used for yogurt preparation as a single starter culture, the time required to reach target titratable acidity (0.9) was 11-12 h. The yogurt fermented with DK211 had favorable panelists ratings for most sensory attributes, which were comparable with yogurt fermented with a commercial strain. The results suggest that strains isolated from kimchi could be potential probiotic and starter cultures for use in yogurt manufacturing.

  16. Lactobacillus priming of the respiratory tract: heterologous immunity and protection against lethal pneumovirus infection

    PubMed Central

    Garcia-Crespo, Katia E.; Chan, Calvin C.; Gabryszewski, Stanislaw J.; Percopo, Caroline M.; Rigaux, Peter; Dyer, Kimberly D.; Domachowske, Joseph B.; Rosenberg, Helene F.

    2013-01-01

    We showed previously that wild-type mice primed via intranasal inoculation with live or heat-inactivated Lactobacillus species were fully (100%) protected against the lethal sequelae of infection with the virulent pathogen, pneumonia virus of mice (PVM), a response that is associated with diminished expression of proinflammatory cytokines and diminished virus recovery. We show here that 40% of the mice primed with live Lactobacillus survived when PVM challenge was delayed for 5 months. This robust and sustained resistance to PVM infection resulting from prior interaction with an otherwise unrelated microbe is a profound example of heterologous immunity. We undertook the present study in order to understand the nature and unique features of this response. We found that intranasal inoculation with L. reuteri elicited rapid, transient neutrophil recruitment in association with proinflammatory mediators (CXCL1, CCL3, CCL2, CXCL10, TNF-alpha and IL-17A) but not Th1 cytokines. IFN? does not contribute to survival promoted by Lactobacillus-priming. Live L. reuteri detected in lung tissue underwent rapid clearance, and was undetectable at 24 hrs after inoculation. In contrast, L. reuteri peptidoglycan (PGN) and L. reuteri genomic DNA (gDNA) were detected at 24 and 48 hours after inoculation, respectively. In contrast to live bacteria, intranasal inoculation with isolated L. reuteri gDNA elicited no neutrophil recruitment, had minimal impact on virus recovery and virus-associated production of CCL3, and provided no protection against the negative sequelae of virus infection. Isolated PGN elicited neutrophil recruitment and proinflammatory cytokines but did not promote sustained survival in response to subsequent PVM infection. Overall, further evaluation of the responses leading to Lactobacillus-mediated heterologous immunity may provide insight into novel antiviral preventive modalities. PMID:23274789

  17. Optimization of bacteriocin production by Lactobacillus sp. MSU3IR against shrimp bacterial pathogens

    PubMed Central

    2013-01-01

    Background Aquaculture is one amongst the growing and major food producing sectors. Shrimp culture is one of the subsectors of aquaculture that attracts more attention because of the economic interest. However, the shrimp culture systems have been facing severe consequences and economical losses due to disease outbreaks. Risk of disease outbreak can be combated with the application of probiotics. For economically viable production of such probiotic products, the present study provides information on the optimization and partial purification of bacteriocin produced by a goat milk isolate Lactobacillus sp. MSU3IR against the shrimp bacterial pathogens. Results Bacteriocin production was estimated as a measure of bactericidal activity (arbitrary Unit/ml) over the test strains. The optimum culture conditions and media components for maximum bacteriocin production by Lactobacillus sp. MSU3IR were: pH: 5.0, temperature: 30°C, carbon source: lactose; nitrogen source: ammonium acetate; NaCl: 3.0% and surfactant: Tween 80. MRS medium was found to extend better bacteriocin production than other tested media. Upon partial purification of bacteriocin, the SDS-PAGE analysis had manifested the presence of two peptide bands with the molecular weight of 39.26 and 6.38 kDa, respectively. Conclusion The present results provide baseline trend for the statistical optimization, scale up process and efficient production of bacteriocin by the candidate bacterial strain Lactobacillus sp. MSU3IR which could be used to replace the usage of conventional chemotherapeutics in shrimp culture systems. PMID:23725298

  18. Application of Lactobacillus immobilized by Activated Carbon Fiber in Fermentation of Lactic Acid in Starch Wastewater

    NASA Astrophysics Data System (ADS)

    Xu, Wei; Wang, Peng; Chi, Guoda; Huang, Chenyong

    2010-11-01

    Activated carbon fibers (ACF) as the carrier of Lactobacillus was introduced into fermenting system, and a method of modifying the surface of ACF by HNO3-Fe (III) was established. Factors that affect ACF carrier's effect on immobilization of Lactobacillus were studied. HCl, H2SO4, HNO3 and FeCl3 solutions were respectively used to modify the surface properties of ACF. The amount of Fe (III) carried on ACF surface was 0.1563 mol/kg after ACF surface was modified by HNO3 for 5 h and then by 0.1 mol/L FeCl3 for 4 h, when the thickness of Lactobacillus on a single silk of carrier reached 40 ?m. When ACF modified by HNO3-Fe (III) was applied in the fermentation of lactic acid in starch industry wastewater, the fermentation period reduced by 8 h and the output of L-lactic acid was 65.5 g/L, which was 3.3% more than that fermented without the carrier.

  19. Lactobacillus buchneri Genotyping on the Basis of Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR) Locus Diversity

    PubMed Central

    Briner, Alexandra E.

    2014-01-01

    Clustered regularly interspaced short palindromic repeats (CRISPR) in combination with associated sequences (cas) constitute the CRISPR-Cas immune system, which uptakes DNA from invasive genetic elements as novel “spacers” that provide a genetic record of immunization events. We investigated the potential of CRISPR-based genotyping of Lactobacillus buchneri, a species relevant for commercial silage, bioethanol, and vegetable fermentations. Upon investigating the occurrence and diversity of CRISPR-Cas systems in Lactobacillus buchneri genomes, we observed a ubiquitous occurrence of CRISPR arrays containing a 36-nucleotide (nt) type II-A CRISPR locus adjacent to four cas genes, including the universal cas1 and cas2 genes and the type II signature gene cas9. Comparative analysis of CRISPR spacer content in 26 L. buchneri pickle fermentation isolates associated with spoilage revealed 10 unique locus genotypes that contained between 9 and 29 variable spacers. We observed a set of conserved spacers at the ancestral end, reflecting a common origin, as well as leader-end polymorphisms, reflecting recent divergence. Some of these spacers showed perfect identity with phage sequences, and many spacers showed homology to Lactobacillus plasmid sequences. Following a comparative analysis of sequences immediately flanking protospacers that matched CRISPR spacers, we identified a novel putative protospacer-adjacent motif (PAM), 5?-AAAA-3?. Overall, these findings suggest that type II-A CRISPR-Cas systems are valuable for genotyping of L. buchneri. PMID:24271175

  20. Probiotic abilities of riboflavin-overproducing Lactobacillus strains: a novel promising application of probiotics.

    PubMed

    Arena, Mattia P; Russo, Pasquale; Capozzi, Vittorio; López, Paloma; Fiocco, Daniela; Spano, Giuseppe

    2014-09-01

    The probiotic potential of Lactobacillus plantarum and Lactobacillus fermentum strains, capable of overproducing riboflavin, was investigated. The riboflavin production was quantified in co-cultures of lactobacilli and human intestinal epithelial cells, and the riboflavin overproduction ability was confirmed. When milk and yogurt were used as carrier matrices, L. plantarum and L. fermentum strains displayed a significant ability to survive through simulated gastrointestinal transit. Adhesion was studied on both biotic and abiotic surfaces. Both strains adhered strongly on Caco-2 cells, negatively influenced the adhesion of Escherichia coli O157:H7, and strongly inhibited the growth of three reference pathogenic microbial strains. Resistance to major antibiotics and potential hemolytic activity were assayed. Overall, this study reveals that these Lactobacillus stains are endowed with promising probiotic properties and thus are candidates for the development of novel functional food which would be both enriched in riboflavin and induce additional health benefits, including a potential in situ riboflavin production, once the microorganisms colonize the host intestine. PMID:24903812

  1. Characterization of the Genome of the Dairy Lactobacillus helveticus Bacteriophage ?AQ113

    PubMed Central

    Scaltriti, Erika; Rossetti, Lia; Guffanti, Alessandro; Armiento, Angelarita; Fornasari, Maria Emanuela; Grolli, Stefano; Carminati, Domenico; Brini, Elena; Pavan, Paolo; Felsani, Armando; D'Urzo, Annalisa; Moles, Anna; Claude, Jean-Baptiste; Grandori, Rita; Ramoni, Roberto; Giraffa, Giorgio

    2013-01-01

    The complete genomic sequence of the dairy Lactobacillus helveticus bacteriophage ?AQ113 was determined. Phage ?AQ113 is a Myoviridae bacteriophage with an isometric capsid and a contractile tail. The final assembled consensus sequence revealed a linear, circularly permuted, double-stranded DNA genome with a size of 36,566 bp and a G+C content of 37%. Fifty-six open reading frames (ORFs) were predicted, and a putative function was assigned to approximately 90% of them. The ?AQ113 genome shows functionally related genes clustered together in a genome structure composed of modules for DNA replication/regulation, DNA packaging, head and tail morphogenesis, cell lysis, and lysogeny. The identification of genes involved in the establishment of lysogeny indicates that it may have originated as a temperate phage, even if it was isolated from natural cheese whey starters as a virulent phage, because it is able to propagate in a sensitive host strain. Additionally, we discovered that the ?AQ113 phage genome is closely related to Lactobacillus gasseri phage KC5a and Lactobacillus johnsonii phage Lj771 genomes. The phylogenetic similarities between L. helveticus phage ?AQ113 and two phages that belong to gut species confirm a possible common ancestral origin and support the increasing consideration of L. helveticus as a health-promoting organism. PMID:23728811

  2. Preparation and application characteristics of microencapsulated Lactobacillus acidophilus as probiotics for dogs.

    PubMed

    Ma, Yuan; Xing, Yage; Xu, Qinglian; Wang, TinXuan; Cai, Yimin; Cao, Dong; Che, Zhengming

    2015-01-01

    In this article, preparation and application characteristics of microencapsulated Lactobacillus acidophilus were investigated. Results indicated that the optimum condition for preparation of micro encapsulation were 10% (w/v) wall material and the temperature of 20°C, respectively. Many micropores in the porous starch micro particles was also observed by Scanning Electron Microscope. Furthermore, the released cell counts were increase from 2.43 log cfu/g to 9.17 log cfu/g for the time prolong to 3h in the simulated colonic pH solution. On the other hand, the visible cells of Lactobacillus acidophilus in the dog feces on the 10th day after the probiotics feeding was improve about 34.8% compare to the before feeding, which was decrease about 24.6%for Escherichia coli. Furthermore, the content of is ovaleric acid, indole and 3-methylindole, putrefactive substances in dog feces, before feeding were reduce 24%, 16% and 45% in dog feces on the 10th day after feeding compared to that before feeding, respectively. Micro encapsulation can be considered a useful technology to provide the protection for Lactobacillus acidophilus and better application effective. PMID:25631500

  3. Development of a Lactobacillus Specific T-RFLP Method to Determine Lactobacilli Diversity in Complex Samples

    PubMed Central

    Chen, Long; Teasdale, Matt T.; Kaczmarczyk, Melissa M.; Freund, Gregory G.; Miller, Michael J.

    2012-01-01

    Terminal restriction fragment length polymorphism (T-RFLP) analysis has been widely used for studying microbial communities. However, most T-RFLP assays use 16S rDNA as the target and are unable to accurately characterize a microbial subpopulation. In this study, we developed a novel T-RFLP protocol based on Lactobacillus hsp60 to rapidly characterize and compare lactobacilli composition. The theoretical terminal restriction fragment (TRF) profiles were calculated from 769 Lactobacillus hsp60 sequences from online databases. In silico digestion with restriction endonucleases AluI and TacI on hsp60 amplicons generated 83 distinct TRF patterns, of which, 70 were species specific. To validate the assay, five previously sequenced lactobacilli were cultured independently, mixed at known concentrations and subjected to analysis by T-RFLP. All five strains generated the predicted TRFs and a qualitative consistent relationship was revealed. We performed the T-RFLP protocol on fecal samples from mice fed 6 different diets (n=4). Principal component analysis and agglomerative hierarchical clustering revealed that the lactobacilli community was strongly connected to dietary supplementation. Our study demonstrates the potential for using Lactobacillus specific T-RFLP to characterize lactobacilli communities in complex samples. PMID:22981747

  4. Preliminary Evaluation of Probiotic Properties of Lactobacillus Strains Isolated from Sardinian Dairy Products

    PubMed Central

    Pisano, Maria Barbara; Viale, Silvia; Conti, Stefania; Deplano, Maura; Melis, Maria Paola; Deiana, Monica

    2014-01-01

    Twenty-three Lactobacillus strains of dairy origin were evaluated for some functional properties relevant to their use as probiotics. A preliminary subtractive screening based on the abilities to inhibit the growth of microbial pathogens and hydrolyze conjugated bile salts was applied, and six strains were selected for further characterization including survival under gastrointestinal environmental conditions, adhesion to gut epithelial tissue, enzymatic activity, and some safety properties. All selected strains maintained elevated cell numbers under conditions simulating passage through the human gastrointestinal tract, well comparable to the values obtained for the probiotic strain Lactobacillus rhamnosus GG, and were able to adhere to Caco-2 cells to various extents (from 3 to 20%). All strains exhibited high aminopeptidase, and absent or very low proteolytic and strong ?-galactosidase activities; none was found to be haemolytic or to produce biogenic amines and all were susceptible to tetracycline, chloramphenicol, erythromycin, ampicillin, and amoxicillin/clavulanic acid. Our results indicate that the Lactobacillus strains analyzed could be considered appropriate probiotic candidates, due to resistance to GIT simulated conditions, antimicrobial activity, adhesion to Caco-2 cell-line, and absence of undesirable properties. They could be used as adjunct cultures for contributing to the quality and health related functional properties of dairy products. PMID:25054135

  5. Obesity-associated gut microbiota is enriched in Lactobacillus reuteri and depleted in Bifidobacterium animalis and Methanobrevibacter smithii

    PubMed Central

    Million, M; Maraninchi, M; Henry, M; Armougom, F; Richet, H; Carrieri, P; Valero, R; Raccah, D; Vialettes, B; Raoult, D

    2012-01-01

    Background: Obesity is associated with increased health risk and has been associated with alterations in bacterial gut microbiota, with mainly a reduction in Bacteroidetes, but few data exist at the genus and species level. It has been reported that the Lactobacillus and Bifidobacterium genus representatives may have a critical role in weight regulation as an anti-obesity effect in experimental models and humans, or as a growth-promoter effect in agriculture depending on the strains. Objectives and methods: To confirm reported gut alterations and test whether Lactobacillus or Bifidobacterium species found in the human gut are associated with obesity or lean status, we analyzed the stools of 68 obese and 47 controls targeting Firmicutes, Bacteroidetes, Methanobrevibacter smithii, Lactococcus lactis, Bifidobacterium animalis and seven species of Lactobacillus by quantitative PCR (qPCR) and culture on a Lactobacillus-selective medium. Findings: In qPCR, B. animalis (odds ratio (OR)=0.63; 95% confidence interval (CI) 0.39–1.01; P=0.056) and M. smithii (OR=0.76; 95% CI 0.59–0.97; P=0.03) were associated with normal weight whereas Lactobacillus reuteri (OR=1.79; 95% CI 1.03–3.10; P=0.04) was associated with obesity. Conclusion: The gut microbiota associated with human obesity is depleted in M. smithii. Some Bifidobacterium or Lactobacillus species were associated with normal weight (B. animalis) while others (L. reuteri) were associated with obesity. Therefore, gut microbiota composition at the species level is related to body weight and obesity, which might be of relevance for further studies and the management of obesity. These results must be considered cautiously because it is the first study to date that links specific species of Lactobacillus with obesity in humans. PMID:21829158

  6. Probiotic potential of lactobacillus strains isolated from sorghum-based traditional fermented food.

    PubMed

    Poornachandra Rao, K; Chennappa, G; Suraj, U; Nagaraja, H; Charith Raj, A P; Sreenivasa, M Y

    2015-06-01

    Sorghum-based traditional fermented food was screened for potential probiotic lactic acid bacteria. The isolates were identified by biochemical, physiological and genetic methods. Species identification was done by 16s rRNA sequence analysis. The functional probiotic potential of the two Lactobacillus species viz., Lactobacillus plantarum (Lact. plantarum) and Lactobacillus pentosus (Lact. pentosus) was assessed by different standard parameters. The strains were tolerant to pH 2 for 1 h and resistant to methicillin, kanamycin, vancomycin and norfloxacin. Two (Lact. plantarum COORG-3 and Lact. pentosus COORG-8) out of eight isolates recorded the cell surface hydrophobicity to be 59.12 and 64.06 %, respectively. All the strains showed tolerance to artificial duodenum juice (pH 2) for 3 h, positive for bile salt hydrolase test and negative for haemolytic test. The neutralized cell-free supernatant of the strains Lact. pentosus COORG-4, Lact. plantarum COORG-1, Lact. plantarum COORG-7, Lact. pentosus COORG-8 and Lact. plantarum COORG-3 showed good antibiofilm activity. Lact. pentosus COORG-8 exhibited 74 % activity against Pseudomonas aeruginosa-MTCC 7903 and Lact. plantarum COORG-7 showed 68 % inhibition of biofilm against Klebsiella pneumonia MTCC 7407. Three (Lact. plantarum COORG-7, Lact. pentosus COORG-5 and Lact. pentosus COORG 8) out of eight isolates exhibited a good antimicrobial activity against Listeria monocytogenes and five isolates (Lact. pentosus COORG 2, Lact. plantarum COORG 1, Lact. plantarum COORG 4, Lact. pentosus COORG 3 and Lact. plantarum COORG 6) are active against Escherichia coli, Bacillus subtilis, Pseudomonas aeruginosa, Enterobacter aerogenes, Klebsiella pneumonia, Enterococcus faecalis. The study also evaluated the cholesterol lowering property of the Lactobacillus strains using hen egg yolk as the cholesterol source. The cholesterol in hen egg yolk was assimilated by 74.12 and 68.26 % by Lact. plantarum COORG 4 and Lact. pentosus COORG 7, respectively. The results of the present study suggest that the Lactobacillus strains isolated and characterized from sorghum-based fermented product may be used as probiotic strains for therapeutic applications. PMID:25666113

  7. Antagonistic effect of Lactobacillus strains against gas-producing coliforms isolated from colicky infants

    PubMed Central

    2011-01-01

    Background Infantile colic is a common disturb within the first 3 months of life, nevertheless the pathogenesis is incompletely understood and treatment remains an open issue. Intestinal gas production is thought to be one of the causes of abdominal discomfort in infants suffering from colic. However, data about the role of the amount of gas produced by infants' colonic microbiota and the correlation with the onset of colic symptoms are scanty. The benefit of supplementation with lactobacilli been recently reported but the mechanisms by which they exert their effects have not yet been fully defined. This study was performed to evaluate the interaction between Lactobacillus spp. strains and gas-forming coliforms isolated from stools of colicky infants. Results Strains of coliforms were isolated from stools of 45 colicky and 42 control breastfed infants in McConkey Agar and identified using PCR with species-specific primers, and the BBL™ Enterotube™ II system for Enterobacteriaceae. Gas-forming capability of coliforms was assessed in liquid cultures containing lactose as sole carbon source. The average count of total coliforms in colicky infants was significantly higher than controls: 5.98 (2.00-8.76) log10 vs 3.90 (2.50-7.10) CFU/g of faeces (p = 0.015). The following strains were identified: Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter aerogenes, Enterobacter cloacae and Enterococcus faecalis. Then, 27 Lactobacillus strains were tested for their antagonistic effect against coliforms both by halo-forming method and in liquid co-cultures. Lactobacillus delbrueckii subsp.delbrueckii DSM 20074 and L. plantarum MB 456 were able to inhibit all coliforms strains (halo-forming method), also in liquid co-cultures, thus demonstrating an antagonistic activity. Conclusions This study shows that two out of 27 strains of Lactobacillus examined possess an antimicrobial effect against six species of gas-forming coliforms isolated from colicky infants. Our findings may stimulate new researches to identify which Lactobacillus strains can improve colicky symptoms by acting on coliforms gut colonization. PMID:21718486

  8. In vitro study of the prebiotic xylooligosaccharide (XOS) on the growth of Bifidobacterium spp and Lactobacillus spp.

    PubMed

    Li, Zhaoping; Summanen, Paula H; Komoriya, Tomoe; Finegold, Sydney M

    2015-12-01

    We recently demonstrated that XOS increased the counts of Bifidobacterium in vivo without increasing Lactobacillus in healthy adults. In the current study, we evaluated the effect of XOS on the growth of 35 Bifidobacterium and 29 Lactobacillus strains in in vitro conditions. Bacteria were identified by 16S rRNA sequence analysis. The growth stimulation was determined by agar dilution technique on plates containing two-fold serial dilutions of XOS (100-0.1?mg/ml). The growth of 86% of Bifidobacterium strains was stimulated at 1.56?mg/ml XOS and 100% at 6.25?mg/ml XOS. The growth of 38% of Lactobacillus strains was stimulated at 1.56?mg/ml XOS and 62% at 6.25?mg/ml XOS; 31% of Lactobacillus were not stimulated by XOS. Our results further suggest that XOS may be beneficial in stimulating intestinal Bifidobacterium without having much effect on Lactobacillus. The potential role for XOS in managing obesity should be investigated further. PMID:26171632

  9. Modulation of cytokine gene expression by selected Lactobacillus isolates in the ileum, caecal tonsils and spleen of Salmonella-challenged broilers.

    PubMed

    Hu, Jie-Lun; Yu, Hai; Kulkarni, Raveendra R; Sharif, Shayan; Cui, Steve W; Xie, Ming-Yong; Nie, Shao-Ping; Gong, Joshua

    2015-12-01

    Probiotics have been used to control Salmonella colonization in the chicken intestine. Recently, we demonstrated that certain selected Lactobacillus isolates were able to reduce Salmonella infection in the chicken spleen and liver as well as down-regulated Salmonella pathogenicity island 1 virulence gene expression in the chicken caecum. To further understand the mechanisms through which Lactobacillus protected chickens from Salmonella infection, the present study has investigated the Lactobacillus isolate(s)-induced host immune response of chickens to Salmonella enterica serovar Typhimurium infection. A thorough examination of cytokine gene expression in the ileum, caecal tonsils, and spleen on days 1 and 3 post-Salmonella infection showed a dynamic spatial and temporal response to Salmonella infection and Lactobacillus treatments. In most instances, it was evident that treatment of chickens with Lactobacillus isolates could significantly attenuate Salmonella-induced changes in the gene expression profile. These included the genes encoding pro-inflammatory cytokines [lipopolysaccharide-induced TNF factor, interleukin (IL)-6, and IL-8], T helper 1 cytokines [IL-12 and interferon (IFN)-?], and T helper 2 cytokines (IL-4 and IL-10). Another important observation from the present investigation was that the response induced by a combination of Lactobacillus isolates was generally more effective than that induced by a single Lactobacillus isolate. Our results show that administration of certain selected Lactobacillus isolates can effectively modulate Salmonella-induced cytokine gene expression, and thus help reduce Salmonella infection in chickens. PMID:26395945

  10. Optimization of antimicrobial substances produced from Lactobacillus paracasei subsp. paracasei NTU 101 (DSM 28047) and Lactobacillus plantarum NTU 102 by response surface methodology.

    PubMed

    Lin, Tzu-Hsing; Pan, Tzu-Ming

    2015-09-01

    Response surface methodology (RSM) was used to study the effects of medium composition and growth conditions on antimicrobial substances produced by Lactobacillus paracasei subsp. paracasei NTU 101 and Lactobacillus plantarum NTU 102. These strains are used in the food and pharmaceutical industries because of their beneficial effects on the human gastrointestinal tract and in immune modulation. The production of antimicrobial substances was optimized by the Box-Behnken experimental design. An empirical model was developed through RSM to describe the relationship between variables (molasses, soymilk, and incubation temperature). The maximum antimicrobial activity of L. paracasei subsp. paracasei NTU 101 was 12.50 AU/mL with 18.33 g/L molasses and 14.53 g/L soymilk at a temperature of 35.00 °C, whereas in MRS (de Man, Rogosa, and Sharpe) broth, the activity was 14.00 AU/mL. Similarly, the maximum antimicrobial activity of L. plantarum NTU 102 was 15.00 AU/mL with 20.56 g/L molasses and 13.54 g/L soymilk at a temperature of 36.50 °C, whereas in MRS, the activity was 13.50 AU/mL. The antimicrobial activities visualized in response surface plots were 12.26 and 15.59 AU/mL for NTU 101 and 102, respectively. The results derived from RSM regression were close to those obtained with the experimental design treatments. These results revealed that an alternative medium could be used for large-scale commercial production of the antimicrobial substances, with potential prospects for application. PMID:26345021

  11. Effects of the Peptide Pheromone Plantaricin A and Cocultivation with Lactobacillus sanfranciscensis DPPMA174 on the Exoproteome and the Adhesion Capacity of Lactobacillus plantarum DC400

    PubMed Central

    Calasso, Maria; Di Cagno, Raffaella; Campanella, Daniela; Minervini, Fabio; Gobbetti, Marco

    2013-01-01

    This study aimed at investigating the extracellular and cell wall-associated proteins (exoproteome) of Lactobacillus plantarum DC400 when cultivated on modified chemically defined medium (CDM) supplemented with the chemically synthesized pheromone plantaricin A (PlnA) or cocultured with L. plantarum DPPMA20 or Lactobacillus sanfranciscensis DPPMA174. Compared to monoculture, two-dimensional gel electrophoresis (2-DE) analysis showed that the exoproteome of L. plantarum DC400 was affected by PlnA and cocultivation with strains DPPMA20 and, especially, DPPMA174. The highest similarity of the 2-DE maps was found between DC400 cells cultivated in monoculture and in coculture with strain DPPMA20. Almost all extracellular proteins (22 spots) and cell wall-associated proteins (40 spots) which showed decreased or increased levels of synthesis during growth in CDM supplemented with PlnA and/or in coculture with strain DPPMA20 or DPPMA174 were identified. On the basis of the sequences in the Kyoto Encyclopedia of Genes and Genomes database, changes to the exoproteome concerned proteins involved in quorum sensing (QS), the transport system, stress response, carbohydrate metabolism and glycolysis, oxidation/reduction processes, the proteolytic system, amino acid metabolism, cell wall and catabolic processes, and cell shape, growth, and division. Cultivation with PlnA and cocultivation with strains DPPMA20 and, especially, DPMMA174 markedly increased the capacity of L. plantarum DC400 to form biofilms, to adhere to human Caco-2 cells, and to prevent the adhesion of potential intestinal pathogens. These phenotypic traits were in part related to oversynthesized moonlighting proteins (e.g., DnaK and GroEL, pyruvate kinase, enolase, and glyceraldehyde-3-phosphate dehydrogenase) in response to QS mechanisms and interaction with L. plantarum DPPMA20 and, especially, L. sanfranciscensis DPPMA174. PMID:23396346

  12. Distinct Immunomodulation of Bone Marrow-Derived Dendritic Cell Responses to Lactobacillus plantarum WCFS1 by Two Different Polysaccharides Isolated from Lactobacillus rhamnosus LOCK 0900

    PubMed Central

    Jachymek, Wojciech; Srutkova, Dagmar; Brzozowska, Ewa; Kozakova, Hana; Gamian, Andrzej

    2014-01-01

    The structures of polysaccharides (PS) isolated from Lactobacillus rhamnosus LOCK 0900 and results from stimulation of mouse bone marrow-derived dendritic cells (BM-DC) and human embryonal kidney (HEK293) cells stably transfected with Toll-like receptors (TLR) upon exposure to these antigens were studied. L. rhamnosus LOCK 0900 produces PS that differ greatly in their structure. The polymer L900/2, with a high average molecular mass of 830 kDa, is a branched heteropolysaccharide with a unique repeating unit consisting of seven sugar residues and pyruvic acid, whereas L900/3 has a low average molecular mass of 18 kDa and contains a pentasaccharide repeating unit and phosphorus. Furthermore, we found that both described PS neither induce cytokine production and maturation of mouse BM-DC nor induce signaling through TLR2/TLR4 receptors. However, they differ profoundly in their abilities to modulate the BM-DC immune response to the well-characterized human isolate Lactobacillus plantarum WCFS1. Exposure to L900/2 enhanced interleukin-10 (IL-10) production induced by L. plantarum WCFS1, while in contrast, L900/3 enhanced the production of IL-12p70. We conclude that PS, probably due to their chemical features, are able to modulate the immune responses to third-party antigens. The ability to induce regulatory IL-10 by L900/2 opens up the possibility to use this PS in therapy of inflammatory conditions, such as inflammatory bowel disease, whereas L900/3 might be useful in reverting the antigen-dependent Th2-skewed immune responses in allergies. PMID:25107979

  13. Method for reliable isolation of Lactobacillus sakei strains originating from Tunisian seafood and meat products.

    PubMed

    Najjari, Afef; Ouzari, Hadda; Boudabous, Abdellatif; Zagorec, Monique

    2008-02-10

    In Tunisia, several food products derived from meat or seafood are naturally processed, without any addition of bacterial starters. Such fermented, dried-cured, salted, or marinated products, as well as the raw meat or fish may thus provide a source to isolate the natural microflora colonizing such environments. We isolated lactic acid bacteria from a representative range of flesh-foods sold or manufactured in different parts of Tunisia, and selectively searched for Lactobacillus sakei, a lactic acid bacterium potentially useful as starter or protective culture. Eighty six (86) strains were isolated from various seafood (anchovy, sardine, sole, mullet, and octopus), or meat (pork, veal, beef, sheep, chicken, and turkey) products that were either fresh, or transformed by different traditional processes. Several methods were used in order to develop a rapid and reliable protocol for the direct identification of L. sakei. Amplified ribosomal DNA restriction analysis (ARDRA) classified the various isolates into 9 distinct groups. Search for the presence of the L. sakei specific katA gene indicated that all positive isolates were grouped in the same ARDRA group. Sequencing of 16S rDNA confirmed those isolates as L. sakei. Those 22 different L. sakei strains represent 25.6% of the total isolates, while other isolates found in the different ARDRA groups were tentatively ascribed to Lactobacillus plantarum, Lactococcus lactis/garviae, Enterococcus avium, Streptococcus parauberis, Hafnia alvei, Pediococcus pentosaceus, and Lactobacillus curvatus through 16S rDNA sequencing. A fast and reliable method to isolate and discriminate L. sakei from complex food environments is proposed. PMID:18155310

  14. Codominance of Lactobacillus plantarum and obligate heterofermentative lactic acid bacteria during sourdough fermentation.

    PubMed

    Ventimiglia, Giusi; Alfonzo, Antonio; Galluzzo, Paola; Corona, Onofrio; Francesca, Nicola; Caracappa, Santo; Moschetti, Giancarlo; Settanni, Luca

    2015-10-01

    Fifteen sourdoughs produced in western Sicily (southern Italy) were analysed by classical methods for their chemico-physical characteristics and the levels of lactic acid bacteria (LAB). pH and total titratable acidity (TTA) were mostly in the range commonly reported for similar products produced in Italy, but the fermentation quotient (FQ) of the majority of samples was above 4.0, due to the low concentration of acetic acid estimated by high performance liquid chromatography (HPLC). Specific counts of LAB showed levels higher than 10(8) CFU g(-1) for many samples. The colonies representing various morphologies were isolated and, after the differentiation based on phenotypic characteristics, divided into 10 groups. The most numerous group was composed of facultative heterofermentative isolates, indicating a relevance of this bacterial group during fermentation. The genetic analysis by randomly amplified polymorphic DNA (RAPD)-PCR, 16S rRNA gene sequencing and species-specific PCRs identified 33 strains as Lactobacillus plantarum, Lactobacillus curvatus and Lactobacillus graminis. Due to the consistent presence of L. plantarum, it was concluded that this species codominates with obligate heterofermentative LAB in sourdough production in this geographical area. In order to evaluate the performances at the basis of their fitness, the 29 L. plantarum strains were investigated for several technological traits. Twelve cultures showed good acidifying abilities in vitro and L. plantarum PON100148 produced the highest concentrations of organic acids. Eleven strains were positive for extracellular protease activity. Bacteriocin-like inhibitory substances (BLIS) production and antifungal activity was scored positive for several strains, included L. plantarum PON100148 which was selected as starter for experimental sourdough production. The characteristics of the sourdoughs and the resulting breads indicated that the best productions were obtained in presence of L. plantarum PON100148. PMID:26187828

  15. Effect of recombinant lactobacillus expressing canine GM-CSF on immune function in dogs.

    PubMed

    Chung, Jin Young; Sung, Eui Jae; Cho, Chun Gyu; Seo, Kyoung Won; Lee, Jong-Soo; Bhang, Dong Ha; Lee, Hee Woo; Hwang, Cheol Yong; Lee, Wan Kyu; Youn, Hwa Young; Kim, Chul Joong

    2009-11-01

    Many Lactobacillus strains have been promoted as good probiotics for the prevention and treatment of diseases. We engineered recombinant Lactobacillus casei, producing biologically active canine granulocyte macrophage colony stimulating factor (cGM-CSF), and investigated its possibility as a good probiotic agent for dogs. Expression of the cGM-CSF protein in the recombinant Lactobacillus was confirmed by SDS-PAGE and Western blotting methods. For the in vivo study, 18 Beagle puppies of 7 weeks of age were divided into three groups; the control group was fed only on a regular diet and the two treatment groups were fed on a diet supplemented with either 1 x 10(9) colony forming units (CFU)/day of L. casei or L. casei expressing cGM-CSF protein for 7 weeks. Body weight was measured, and fecal and blood samples were collected from the dogs during the experiment for the measurement of hematology, fecal immunoglobulin (Ig)A and IgG, circulating IgA and IgG, and canine corona virus (CCV)-specific IgG. There were no differences in body weights among the groups, but monocyte counts in hematology and serum IgA were higher in the group receiving L. casei expressing cGMCSF than in the other two groups. After the administration of CCV vaccine, CCV-specific IgG in serum increased more in the group supplemented with L. casei expressing cGM-CSF than the other two groups. This study shows that a dietary L. casei expressing cGM-CSF enhances specific immune functions at both the mucosal and systemic levels in puppies. PMID:19996694

  16. Diversity and metabolic impact of intestinal Lactobacillus species in healthy adults and the elderly.

    PubMed

    Štšepetova, Jelena; Sepp, Epp; Kolk, Helgi; Lõivukene, Krista; Songisepp, Epp; Mikelsaar, Marika

    2011-04-01

    The present study aimed at assessing the counts and species distribution of intestinal lactobacilli and exploring if the data are associated with BMI and blood glucose level in healthy adults and elderly persons. The BMI (P < 0·01), the level of fasting blood glucose (P < 0·001) and the total counts of lactobacilli (P < 0·01 by bacteriology; P < 0·001 by real-time PCR) were higher in the elderly. The number of species in adults was lower (P < 0·05), who were more often colonised with Lactobacillus acidophilus (P = 0·031) and L. helveticus (P < 0·001). In contrast, L. plantarum (P = 0·035), L. paracasei (P < 0·001) and L. reuteri (P = 0·031) were more prevalent in the elderly. L. rhamnosus was detected in adults (P < 0·001), but not in any elderly person. BMI was associated with counts of lactobacilli, adjusted for age and sex (P = 0·008). The higher BMI in both groups of persons was associated with the presence of obligate homofermentative lactobacilli and L. sakei, both adjusted for age and sex. Plasma glucose values were positively correlated with BMI and negatively correlated with colonisation with L. paracasei (P = 0·0238) in adults and on the borderline with L. fermentum (P = 0·052) in the elderly. Thus, the species-specific PCR analysis of Lactobacillus sp. combined with viable plating data indicates substantial age-related structural differences in the intestinal lactobacilli communities. The higher counts of intestinal Lactobacillus sp. are associated with higher BMI and blood glucose content, while their specific fermentative groups and species of lactobacilli appear at different glucose levels both in adults and in the elderly. PMID:21303568

  17. Evaluation of the probiotic properties of new Lactobacillus and Bifidobacterium strains and their in vitro effect.

    PubMed

    Presti, I; D'Orazio, G; Labra, M; La Ferla, B; Mezzasalma, V; Bizzaro, G; Giardina, S; Michelotti, A; Tursi, F; Vassallo, M; Di Gennaro, P

    2015-07-01

    Probiotic ingestion is recommended as a preventive approach to maintain the balance of the intestinal microbiota and to enhance the human well-being. During the whole life of each individual, the gut microbiota composition could be altered by lifestyle, diet, antibiotic therapies and other stress conditions, which may lead to acute and chronic disorders. Hence, probiotics can be administered for the prevention or treatment of some disorders, including lactose malabsorption, acute diarrhoea, irritable bowel syndrome, necrotizing enterocolitis and mild forms of inflammatory bowel disease. The probiotic-mediated effect is an important issue that needs to be addressed in relation to strain-specific probiotic properties. In this work, the probiotic properties of new Lactobacillus and Bifidobacterium strains were screened, and their effects in vitro were evaluated. They were screened for probiotic properties by determining their tolerance to low pH and to bile salts, antibiotic sensitivity, antimicrobial activity and vitamin B8, B9 and B12 production, and by considering their ability to increase the antioxidant potential and to modulate the inflammatory status of systemic-miming cell lines in vitro. Three out of the examined strains presenting the most performant probiotic properties, as Lactobacillus plantarum PBS067, Lactobacillus rhamnosus PBS070 and Bifidobacterium animalis subsp. lactis PBSO75, were evaluated for their effects also on human intestinal HT-29 cell line. The obtained results support the possibility to move to another level of study, that is, the oral administration of these probiotical strains to patients with acute and chronic gut disorders, by in vivo experiments. PMID:25744647

  18. Complete genome sequence of Lactobacillus heilongjiangensis DSM 28069(T): Insight into its probiotic potential.

    PubMed

    Zheng, Beiwen; Jiang, Xiawei; Cheng, Hong; Xu, Zemin; Li, Ang; Hu, Xinjun; Xiao, Yonghong

    2015-12-20

    Lactobacillus heilongjiangensis DSM 28069(T) is a potential probiotic isolated from traditional Chinese pickle. Here we report the complete genome sequence of this strain. The complete genome is 2,790,548bp with the GC content of 37.5% and devoid of plasmids. Sets of genes involved in the biosynthesis of riboflavin and folate were identified in the genome, which revealed its potential application in biotechnological industry. The genome sequence of L. heilongjiangensis DSM 28069(T) now provides the fundamental information for future studies. PMID:26432337

  19. Antilisterial Bacteriocin from Lactobacillus rhamnosus CJNU 0519 Presenting a Narrow Antimicrobial Spectrum

    PubMed Central

    2015-01-01

    A lactic acid bacterium presenting antimicrobial activity against a Lactobacillus acidophilus strain used for eradication of acid inhibition was isolated from a natural cheese. The 16S rRNA gene sequence of the isolate best matched with a strain of L. rhamnosus and was designated L. rhamnosus CJNU 0519. The antimicrobial activity of the partially purified bacteriocin of CJNU 0519 was abolished when treated with a protease, indicating the protein nature of the bacteriocin. The partially purified bacteriocin (rhamnocin 519) displayed a narrow antimicrobial activity against L. acidophilus, Listeria monocytogenes, and Staphylococcus aureus among several tested bacterial and yeast strains. Rhamnocin 519 in particular showed strong bactericidal action against L. monocytogenes.

  20. Immune Regulatory Effect of Newly Isolated Lactobacillus delbrueckii from Indian Traditional Yogurt.

    PubMed

    Hong, Yi-Fan; Lee, Yoon-Doo; Park, Jae-Yeon; Jeon, Boram; Jagdish, Deepa; Jang, Soojin; Chung, Dae Kyun; Kim, Hangeun

    2015-08-01

    Lactic acid bacteria (LAB) are microorganisms that are believed to provide health benefits. Here, we isolated LAB from Indian fermented foods, such as traditional Yogurt and Dosa. LAB from Yogurt most significantly induced TNF-? and IL-1? production, whereas LAB from Dosa induced mild cytokine production. After 16S rRNA gene sequencing and phylogenetic analysis, a Yogurt-borne lactic acid bacterium was identified and classified as Lactobacillus delbrueckii subsp. bulgaricus, and it was renamed L. delbrueckii K552 for the further studies. Our data suggest that the newly isolated L. delbrueckii can be used for the treatment of immune deficiency disorders. PMID:25839333

  1. Evaluation of stored lamb bio-preserved using a three-strain cocktail of Lactobacillus sakei.

    PubMed

    Jones, R J; Wiklund, E; Zagorec, M; Tagg, J R

    2010-12-01

    Commercially prepared lamb was stored at -1.5 °C after inoculation with a combination of three Lactobacillus sakei strains previously shown to inhibit spoilage and pathogenic bacteria of importance to the meat industry. Between 6 and 14 weeks storage samples were evaluated for growth of inoculated strains, production of fermentation end-products and sensory acceptance of the cooked product. All three L. sakei strains flourished during storage, formed consistently dominant populations and were associated with lower surface pH and increased levels of lactic and acetic acids. Inoculated samples were determined to be as equally acceptable for smell, acidity, rancidity and overall liking as un-inoculated controls. PMID:20800371

  2. Genomic diversity and versatility of Lactobacillus plantarum, a natural metabolic engineer.

    PubMed

    Siezen, Roland J; van Hylckama Vlieg, Johan E T

    2011-08-30

    In the past decade it has become clear that the lactic acid bacterium Lactobacillus plantarum occupies a diverse range of environmental niches and has an enormous diversity in phenotypic properties, metabolic capacity and industrial applications. In this review, we describe how genome sequencing, comparative genome hybridization and comparative genomics has provided insight into the underlying genomic diversity and versatility of L. plantarum. One of the main features appears to be genomic life-style islands consisting of numerous functional gene cassettes, in particular for carbohydrates utilization, which can be acquired, shuffled, substituted or deleted in response to niche requirements. In this sense, L. plantarum can be considered a "natural metabolic engineer". PMID:21995294

  3. Brewer's spent grain as raw material for lactic acid production by Lactobacillus delbrueckii.

    PubMed

    Mussatto, Solange I; Fernandes, Marcela; Dragone, Giuliano; Mancilha, Ismael M; Roberto, Inês C

    2007-12-01

    Chemically pre-treated brewer's spent grain was saccharified with cellulase producing a hydrolysate with approx. 50 g glucose l(-1). This hydrolysate was used as a fermentation medium without any nutrient supplementation by Lactobacillus delbrueckii, which produced L-lactic acid (5.4 g l(-1)) at 0.73 g g(-1) glucose consumed (73% efficiency). An inoculum of 1 g dry cells l(-1) gave the best yield of the process, but the pH decrease affected the microorganism capacity to consume glucose and convert it into lactic acid. PMID:17700998

  4. Reutericyclin producing Lactobacillus reuteri modulates development of fecal microbiota in weanling pigs

    PubMed Central

    Yang, Yan; Zhao, Xin; Le, Minh H. A.; Zijlstra, Ruurd T.; Gänzle, Michael G.

    2015-01-01

    Lactobacillus reuteri is used as probiotic culture in food and feed applications; however, strain specific properties of L. reuteri that mediate probiotic activity remain unknown. This study aimed to determine effects of feed fermentation with exopolysaccharide and reutericyclin producing L. reuteri on the transition of the gut microbiome of piglets after weaning. The reutericyclin and reuteran producing L. reuteri TMW1.656 was compared to the reutericyclin negative and levan producing L. reuteri LTH5794 and unfermented controls. Both strains were fermented at conditions supporting exopolysaccharide formation, or at conditions not supporting exopolysaccharide formation. Fecal microbiota were characterized by partial sequencing of 16S rRNA genes, and by quantitative PCR targeting clostridial toxins. The transition to solid food resulted in a transient increase of Proteobacteria to 12% of total bacteria, and increased bacterial diversity by increasing the abundance of anaerobic fiber fermenting Firmicutes. Three weeks after weaning, Prevotella and Lactobacillus were among the dominant bacterial genera. Feed fermentation with L. reuteri affected the abundance of few bacterial taxa and particularly reduced the abundance of Enterobacteriaceae (P < 0.05) when compared to unfermented controls. Reutericyclin producing L. reuteri increased the abundance of Dialister spp. and Mitsuokella spp. (P < 0.05) but did not influence the abundance of clostridial toxins in the feces. In conclusion, data on the contribution of specific metabolic activities of L. reuteri to probiotic activity will facilitate the strain selection for probiotic applications in food and feed. PMID:26284047

  5. Expression of FABP4, adipsin and adiponectin in Paneth cells is modulated by gut Lactobacillus

    PubMed Central

    Su, Xiaomin; Yan, Hui; Huang, Yugang; Yun, Huan; Zeng, Benhua; Wang, Enlin; Liu, Yu; Zhang, Yuan; Liu, Feifei; Che, Yongzhe; Zhang, Zhiqian; Yang, Rongcun

    2015-01-01

    We here found that intestinal epithelial Paneth cells secrete FABP4, adipsin and adiponectin in both mice and human. Deletion of Paneth cell results in the decrease of FABP4, adipsin and adiponectin not only in intestinal crypt cells but also in sera, suggesting that they may influence the state of the whole body. We also demonstrate that expression of FABP4, adipsin and adiponectin may be modulated by specific gut microbiota. In germ-free (GF) mice, the expression of FABP4, adipsin and adiponectin were lower or difficult to be detected. Feces transplantation promoted the expression of FABP4, adipsin and adiponectin in gut epithelial Paneth cells. We have found that Lactobacillus NK6 colony, which has the highest similarity with Lactobacillus taiwanensis strain BCRC 17755, may induce the expression of FABP4, adipsin and adiponectin through TRAF2 and TRAF6 ubiquitination mediated NF-?B signaling. Taken together, our findings set up a novel mechanism for FABP4, adipsin and adiponectin through gut microbiota mediating expression in gut Paneth cells. PMID:26687459

  6. Dietary supplementation with lactose or artificial sweetener enhances swine gut Lactobacillus population abundance.

    PubMed

    Daly, Kristian; Darby, Alistair C; Hall, Neil; Nau, Alexandra; Bravo, David; Shirazi-Beechey, Soraya P

    2014-06-01

    The commensal bacteria Lactobacillus are widely used as probiotic organisms conferring a heath benefit on the host. They have been implicated in promoting gut health via the stimulation of host immunity and anti-inflammatory responses, as well as protecting the intestinalmucosa against pathogen invasion. Lactobacilli grow by fermenting sugars and starches and produce lactic acid as their primary metabolic product. For efficient utilisation of varied carbohydrates, lactobacilli have evolved diverse sugar transport and metabolic systems, which are specifically induced by their own substrates. Many bacteria are also capable of sensing and responding to changes in their environment. These sensory responses are often independent of transport or metabolism and are mediated through membrane-spanning receptor proteins. We employed DNA-based pyrosequencing technology to investigate the changes in the intestinal microbiota of piglets weaned to a diet supplemented with either a natural sugar, lactose or an artificial sweetener (SUCRAM®, consisting of saccharin and neohesperidin dihydrochalcone (NHDC); Pancosma SA). The addition of either lactose or saccharin/NHDC to the piglets' feed dramatically increased the caecal population abundance of Lactobacillus, with concomitant increases in intraluminal lactic acid concentrations. This is the first report of the prebiotic-like effects of saccharin/NHDC, an artificial sweetener, being able to influence the commensal gut microbiota. The identification of the underlying mechanism(s) will assist in designing nutritional strategies for enhancing gut immunity and maintaining gut health. PMID:24382146

  7. Viability and growth characteristics of Lactobacillus in soymilk supplemented with B-vitamins.

    PubMed

    Ewe, Joo-Ann; Wan-Abdullah, Wan-Nadiah; Liong, Min-Tze

    2010-02-01

    Ten strains of Lactobacillus were evaluated for their viability in soymilk. Lactobacillus acidophilus ATCC 314, L. acidophilus FTDC 8833, L. acidophilus FTDC 8633 and L. gasseri FTDC 8131 displayed higher viability in soymilk and were thus selected to be evaluated for viability and growth characteristics in soymilk supplemented with B-vitamins. Pour plate analyses showed that the supplementation of all B-vitamins studied promoted the growth of lactobacilli to a viable count exceeding 7 log CFU/ml. alpha-Galactosidase specific activity of lactobacilli as determined spectrophotometrically showed an increase upon supplementation of B-vitamins. High-performance liquid chromatography analyses revealed that this led to increased hydrolysis of soy oligosaccharides and subsequently higher utilization of simple sugars. Production of organic acids as determined via high-performance liquid chromatography also showed an increase, accompanied by a decrease in pH of soymilk. Additionally, the supplementation of B-vitamins also promoted the synthesis of riboflavin and folic acid by lactobacilli in soymilk. Our results indicated that B-vitamin-supplemented soymilk is a good proliferation medium for strains of lactobacilli. PMID:19961357

  8. Generation of food-grade recombinant Lactobacillus casei delivering Myxococcus xanthus prolyl endopeptidase

    PubMed Central

    Alvarez-Sieiro, Patricia; Martin, Maria Cruz; Redruello, Begoña; del Rio, Beatriz; Ladero, Victor; Palanski, Brad A.; Khosla, Chaitan; Fernandez, Maria; Alvarez, Miguel A.

    2015-01-01

    Prolyl endopeptidases (PEP), a family of serine proteases with the ability to hydrolyze the peptide bond on the carboxyl side of an internal proline residue, are able to degrade immunotoxic peptides responsible for celiac disease (CD), such as a 33-residue gluten peptide (33-mer). Oral administration of PEP has been suggested as a potential therapeutic approach for CD, although delivery of the enzyme to the small intestine requires intrinsic gastric stability or advanced formulation technologies. We have engineered two food-grade Lactobacillus casei strains to deliver PEP in an in vitro model of small intestine environment. One strain secretes PEP into the extracellular medium, whereas the other retains PEP in the intracellular environment. The strain that secretes PEP into the extracellular medium is the most effective to degrade the 33-mer and is resistant to simulated gastrointestinal stress. Our results suggest that in a future, after more studies and clinical trials, an engineered food-grade Lactobacillus strain may be useful as a vector for in situ production of PEP in the upper small intestine of CD patients. PMID:24752841

  9. Comparative proteomic analysis of biofilm and planktonic cells of Lactobacillus plantarum DB200.

    PubMed

    De Angelis, Maria; Siragusa, Sonya; Campanella, Daniela; Di Cagno, Raffaella; Gobbetti, Marco

    2015-07-01

    This study investigated the relative abundance of extracellular and cell wall associated proteins (exoproteome), cytoplasmic proteins (proteome), and related phenotypic traits of Lactobacillus plantarum grown under planktonic and biofilm conditions. Lactobacillus plantarum DB200 was preliminarily selected due to its ability to form biofilms and to adhere to Caco2 cells. As shown by fluorescence microscope analysis, biofilm cells became longer and autoaggregated at higher levels than planktonic cells. The molar ratio between glucose consumed and lactate synthesised was markedly decreased under biofilm compared to planktonic conditions. DIGE analysis showed a differential exoproteome (115 protein spots) and proteome (44) between planktonic and biofilm L. plantarum DB200 cells. Proteins up- or downregulated by at least twofold (p < 0.05) were found to belong mainly to the following functional categories: cell wall and catabolic process, cell cycle and adhesion, transport, glycolysis and carbohydrate metabolism, exopolysaccharide metabolism, amino acid and protein metabolisms, fatty acid and lipid biosynthesis, purine and nucleotide metabolism, stress response, oxidation/reduction process, and energy metabolism. Many of the above proteins showed moonlighting behavior. In accordance with the high expression levels of stress proteins (e.g., DnaK, GroEL, ClpP, GroES, and catalase), biofilm cells demonstrated enhanced survival under conditions of environmental stress. PMID:25728239

  10. Detection and Identification of Probiotic Lactobacillus plantarum Strains by Multiplex PCR Using RAPD-Derived Primers.

    PubMed

    Galanis, Alex; Kourkoutas, Yiannis; Tassou, Chrysoula C; Chorianopoulos, Nikos

    2015-01-01

    Lactobacillus plantarum 2035 and Lactobacillus plantarum ACA-DC 2640 are two lactic acid bacteria (LAB) strains that have been isolated from Feta cheese. Both display significant potential for the production of novel probiotic food products. The aim of the present study was the development of an accurate and efficient method for the molecular detection and identification of the above strains in a single reaction. A multiplex PCR assay was designed for each strain, based on specific primers derived from Random Amplified Polymorphic DNA (RAPD) Sequenced Characterized Amplified Region (SCAR) analysis. The specificity of the assay was tested with a total of 23 different LAB strains, for L. plantarum 2035 and L. plantarum ACA-DC 2640. The multiplex PCR assay was also successfully applied for the detection of the above cultures in yogurt samples prepared in our lab. The proposed methodology may be applied for monitoring the presence of these strains in food products, thus evaluating their probiotic character. Moreover, our strategy may be adapted for other novel LAB strains with probiotic potential, thus providing a powerful tool for molecular discrimination that could be invaluable to the food industry. PMID:26506345

  11. Probiotic Potential of Lactobacillus Strains with Antifungal Activity Isolated from Animal Manure

    PubMed Central

    Ilavenil, Soundharrajan; Park, Hyung Soo; Vijayakumar, Mayakrishnan; Valan Arasu, Mariadhas; Kim, Da Hye; Ravikumar, Sivanesan; Choi, Ki Choon

    2015-01-01

    The aim of the study was to isolate and characterize the lactic acid bacteria (LAB) from animal manure. Among the thirty LAB strains, four strains, namely, KCC-25, KCC-26, KCC-27, and KCC-28, showed good cell growth and antifungal activity and were selected for further characterization. Biochemical and physiology properties of strains confirmed that the strains are related to the Lactobacillus sp.; further, the 16S rRNA sequencing confirmed 99.99% sequence similarity towards Lactobacillus plantarum. The strains exhibited susceptibility against commonly used antibiotics with negative hemolytic property. Strains KCC-25, KCC-26, KCC-27, and KCC-28 showed strong antifungal activity against Aspergillus fumigatus, Penicillium chrysogenum, Penicillium roqueforti, Botrytis elliptica, and Fusarium oxysporum, respectively. Fermentation studies noted that the strains were able to produce significant amount of lactic, acetic, and succinic acids. Further, the production of extracellular proteolytic and glycolytic enzymes, survival under low pH, bile salts, and gastric juice together with positive bile salt hydrolase (Bsh) activity, cholesterol lowering, cell surface hydrophobicity, and aggregation properties were the strains advantages. Thus, KCC-25, KCC-26, KCC-27, and KCC-28 could have the survival ability in the harsh condition of the digestive system in the gastrointestinal tract. In conclusion, novel L. plantarum KCC-25, KCC-26, KCC-27, and KCC-28 could be considered as potential antimicrobial probiotic strains. PMID:26167534

  12. The inhibitory effect of a Lactobacillus acidophilus derived biosurfactant on biofilm producer Serratia marcescens

    PubMed Central

    Shokouhfard, Maliheh; Kermanshahi, Rouha Kasra; Shahandashti, Roya Vahedi; Feizabadi, Mohammad Mehdi; Teimourian, Shahram

    2015-01-01

    Objective(s): Serratia marcescens is one of the nosocomial pathogen with the ability to form biofilm which is an important feature in the pathogenesis of S. marcescens. The aim of this study was to determine the anti-adhesive properties of a biosurfactant isolated from Lactobacillus acidophilus ATCC 4356, on S. marcescens strains. Materials and Methods: Lactobacillus acidophilus ATCC 4356 was selected as a probiotic strain for biosurfactant production. Anti-adhesive activities was determined by pre-coating and co- incubating methods in 96-well culture plates. Results: The FTIR analysis of derived biosurfactant revealed the composition as protein component. Due to the release of such biosurfactants, L. acidophilus was able to interfere with the adhesion and biofilm formation of the S. marcescens strains. In co-incubation method, this biosurfactant in 2.5 mg/ml concentration showed anti-adhesive activity against all tested strains of S. marcescens (P<0.05). Conclusion: Our results show that the anti-adhesive properties of L. acidophilus biosurfactant has the potential to be used against microorganisms responsible for infections in the urinary, vaginal and gastrointestinal tracts, as well as skin, making it a suitable alternative to conventional antibiotics.

  13. High-resolution structures of Lactobacillus salivarius transketolase in the presence and absence of thiamine pyrophosphate

    PubMed Central

    Lukacik, Petra; Lobley, Carina M. C.; Bumann, Mario; Arena de Souza, Victoria; Owens, Raymond J.; O’Toole, Paul W.; Walsh, Martin A.

    2015-01-01

    Probiotic bacterial strains have been shown to enhance the health of the host through a range of mechanisms including colonization, resistance against pathogens, secretion of antimicrobial compounds and modulation of the activity of the innate immune system. Lactobacillus salivarius UCC118 is a well characterized probiotic strain which survives intestinal transit and has many desirable host-interaction properties. Probiotic bacteria display a wide range of catabolic activities, which determine their competitiveness in vivo. Some lactobacilli are heterofermentative and can metabolize pentoses, using a pathway in which transketolase and transaldolase are key enzymes. L. salivarius UCC118 is capable of pentose utilization because it encodes the key enzymes on a megaplasmid. The crystal structures of the megaplasmid-encoded transketolase with and without the enzyme cofactor thiamine pyrophosphate have been determined. Comparisons with other known transketolase structures reveal a high degree of structural conservation in both the catalytic site and the overall conformation. This work extends structural knowledge of the transketolases to the industrially and commercially important Lactobacillus genus. PMID:26457526

  14. Impact of kefir derived Lactobacillus kefiri on the mucosal immune response and gut microbiota.

    PubMed

    Carasi, P; Racedo, S M; Jacquot, C; Romanin, D E; Serradell, M A; Urdaci, M C

    2015-01-01

    The evaluation of the impact of probiotics on host health could help to understand how they can be used in the prevention of diseases. On the basis of our previous studies and in vitro assays on PBMC and Caco-2 ccl20:luc reporter system presented in this work, the strain Lactobacillus kefiri CIDCA 8348 was selected and administrated to healthy Swiss mice daily for 21 days. The probiotic treatment increased IgA in feces and reduced expression of proinflammatory mediators in Peyer Patches and mesenteric lymph nodes, where it also increased IL-10. In ileum IL-10, CXCL-1 and mucin 6 genes were upregulated; meanwhile in colon mucin 4 was induced whereas IFN-?, GM-CSF, and IL-1? genes were downregulated. Moreover, ileum and colon explants showed the anti-inflammatory effect of L. kefiri since the LPS-induced increment of IL-6 and GM-CSF levels in control mice was significantly attenuated in L. kefiri treated mice. Regarding fecal microbiota, DGGE profiles allowed differentiation of experimental groups in two separated clusters. Quantitative PCR analysis of different bacterial groups revealed only significant changes in Lactobacillus population. In conclusion, L. kefiri is a good candidate to be used in gut inflammatory disorders. PMID:25811034

  15. Adhesion of indigenous Lactobacillus plantarum to gut extracellular matrix and its physicochemical characterization.

    PubMed

    Yadav, Ashok Kumar; Tyagi, Ashish; Kumar, Ashwani; Saklani, Asha Chandola; Grover, Sunita; Batish, Virender Kumar

    2015-03-01

    Adhesion to the human intestinal epithelial cell is considered as one of the important selection criteria of lactobacilli for probiotic attributes. Sixteen Lactobacillus plantarum strains from human origins were subjected for adhesion to extracellular matrix (ECM) components, and their physiochemical characterization, incubation time course and effect of different pH on bacterial adhesion in vitro were studied. Four strains showed significant binding to both fibronectin and mucin. After pretreatment with pepsin and trypsin, the bacterial adhesion to ECM reduced to the level of 50 % and with lysozyme significantly decreased by 65-70 %. Treatment with LiCl also strongly inhibited (90 %) the bacterial adhesion to ECM. Tested strains showed highest binding efficacy at time course of 120 and 180 min. Additionally, the binding of Lp91 to ECM was highest at pH 6 (155 ± 2.90 CFU/well). This study proved that surface layer components are proteinaceous in nature, which contributed in adhesion of lactobacillus strains. Further, the study can provide a better platform for introduction of new indigenous probiotic strains having strong adhesion potential for future use. PMID:25212764

  16. Lactobacillus protects the integrity of intestinal epithelial barrier damaged by pathogenic bacteria.

    PubMed

    Yu, Qinghua; Yuan, Lixia; Deng, Jun; Yang, Qian

    2015-01-01

    Pathogens invade intestinal mucosal barrier through phagocytosis of antigen presenting cells (dendritic cell, microfold cells), or through the invasion into the intestinal epithelial directly. Some pathogens could damage the cell junction between epithelial cells and use the paracellular pathway as an entrance to invade. Moreover, some Lactobacillus could inhibit the adhesion of the pathogens and protect the integrity of the cell junction and mucosal barrier. This research focused on the potential therapeutic effect of Lactobacillus fructosus (L. fructosus) C2 to attenuate ETEC K88 or S. typhimurium SL1344 induced changes to mucosal barrier. The results demonstrated that treatment of polarized Caco-2 cells with L. fructosus C2 reduced the permeation of dextran, and expression of IL-8, p-ERK, and p-JNK when cells were infected with pathogenic bacteria. The findings indicated that L. fructosus C2 exerted a protective effect against the damage to the integrity of Caco-2 cells by ETEC or S. typhimurium infection. PMID:25859435

  17. In situ examination of Lactobacillus brevis after exposure to an oxidizing disinfectant

    PubMed Central

    Zhao, Yu; Knøchel, Susanne; Siegumfeldt, Henrik

    2014-01-01

    Beer is a hostile environment for most microorganisms, but some lactic acid bacteria can grow in this environment. This is primarily because these organisms have developed the ability to grow in the presence of hops. It has been speculated that hop resistance is inversely correlated to resistance against oxidation, and this would have great impact on the use of various disinfectants in the brewing industry. In this study, we cultivated bacteria under aerobic and anaerobic conditions, and then investigated the in situ outgrowth of individual cells into microcolonies on de Man Rogosa Sharpe (MRS) agar after exposure to the oxidizing agent peracetic acid (PAA). An automated microscope stage allowed us to analyse a much larger number of cells over extended periods of incubation. After PAA treatment, the lag time increased markedly, and extensive variation in morphology, ?max as well as stress resistance was observed between and within the tested Lactobacillus brevis strains. The results suggest that aerobic cultivation increased the oxidative stress tolerance in Lactobacillus brevis. The results also show that dead cells are randomly distributed in a microcolony and the majority of non-growing individual cells do not stain with a membrane impermanent dye (Propidium iodide), which indicates that PAA may not destroy the plasma membrane. In conclusion, the developed microscopic analysis of individual cells on MRS agar can provides faster results and more details of cell physiology compared to the traditional CFU method. PMID:25505451

  18. In situ examination of Lactobacillus brevis after exposure to an oxidizing disinfectant.

    PubMed

    Zhao, Yu; Knøchel, Susanne; Siegumfeldt, Henrik

    2014-01-01

    Beer is a hostile environment for most microorganisms, but some lactic acid bacteria can grow in this environment. This is primarily because these organisms have developed the ability to grow in the presence of hops. It has been speculated that hop resistance is inversely correlated to resistance against oxidation, and this would have great impact on the use of various disinfectants in the brewing industry. In this study, we cultivated bacteria under aerobic and anaerobic conditions, and then investigated the in situ outgrowth of individual cells into microcolonies on de Man Rogosa Sharpe (MRS) agar after exposure to the oxidizing agent peracetic acid (PAA). An automated microscope stage allowed us to analyse a much larger number of cells over extended periods of incubation. After PAA treatment, the lag time increased markedly, and extensive variation in morphology, ?max as well as stress resistance was observed between and within the tested Lactobacillus brevis strains. The results suggest that aerobic cultivation increased the oxidative stress tolerance in Lactobacillus brevis. The results also show that dead cells are randomly distributed in a microcolony and the majority of non-growing individual cells do not stain with a membrane impermanent dye (Propidium iodide), which indicates that PAA may not destroy the plasma membrane. In conclusion, the developed microscopic analysis of individual cells on MRS agar can provides faster results and more details of cell physiology compared to the traditional CFU method. PMID:25505451

  19. Impact of Kefir Derived Lactobacillus kefiri on the Mucosal Immune Response and Gut Microbiota

    PubMed Central

    Carasi, P.; Racedo, S. M.; Jacquot, C.; Romanin, D. E.; Serradell, M. A.; Urdaci, M. C.

    2015-01-01

    The evaluation of the impact of probiotics on host health could help to understand how they can be used in the prevention of diseases. On the basis of our previous studies and in vitro assays on PBMC and Caco-2 ccl20:luc reporter system presented in this work, the strain Lactobacillus kefiri CIDCA 8348 was selected and administrated to healthy Swiss mice daily for 21 days. The probiotic treatment increased IgA in feces and reduced expression of proinflammatory mediators in Peyer Patches and mesenteric lymph nodes, where it also increased IL-10. In ileum IL-10, CXCL-1 and mucin 6 genes were upregulated; meanwhile in colon mucin 4 was induced whereas IFN-?, GM-CSF, and IL-1? genes were downregulated. Moreover, ileum and colon explants showed the anti-inflammatory effect of L. kefiri since the LPS-induced increment of IL-6 and GM-CSF levels in control mice was significantly attenuated in L. kefiri treated mice. Regarding fecal microbiota, DGGE profiles allowed differentiation of experimental groups in two separated clusters. Quantitative PCR analysis of different bacterial groups revealed only significant changes in Lactobacillus population. In conclusion, L. kefiri is a good candidate to be used in gut inflammatory disorders. PMID:25811034

  20. Intragastric and Intranasal Administration of Lactobacillus paracasei NCC2461 Modulates Allergic Airway Inflammation in Mice

    PubMed Central

    Pellaton, Céline; Nutten, Sophie; Thierry, Anne-Christine; Boudousquié, Caroline; Barbier, Nathalie; Blanchard, Carine; Corthésy, Blaise; Mercenier, Annick; Spertini, François

    2012-01-01

    Introduction. Preclinical and clinical evidences for a role of oral probiotics in the management of allergic diseases are emerging. Aim. We aimed at testing the immunomodulatory effects of intranasal versus intragastric administration of Lactobacillus paracasei NCC2461 in a mouse model of allergic airway inflammation and the specificity of different probiotics by comparing L. paracasei NCC2461 to Lactobacillus plantarum NCC1107. Methods. L. paracasei NCC2461 or L. plantarum NCC1107 strains were administered either intragastrically (NCC2461) or intranasally (NCC2461 or NCC1107) to OVA-sensitized mice challenged with OVA aerosols. Inflammatory cell recruitment into BALF, eotaxin and IL-5 production in the lungs were measured. Results. Intranasal L. paracasei NCC2461 efficiently protected sensitized mice upon exposure to OVA aerosols in a dose-dependent manner as compared to control mice. Inflammatory cell number, eotaxin and IL-5 were significantly reduced in BALF. Intranasal supplementation of L. paracasei NCC2461 was more potent than intragastric application in limiting the allergic response and possibly linked to an increase in T regulatory cells in the lungs. Finally, intranasal L. plantarum NCC1107 reduced total and eosinophilic lung inflammation, but increased neutrophilia and macrophages infiltration. Conclusion. A concerted selection of intervention schedule, doses, and administration routes (intranasal versus intragastric) may markedly contribute to modulate airway inflammation in a probiotic strain-specific manner. PMID:22762009

  1. Intragastric and Intranasal Administration of Lactobacillus paracasei NCC2461 Modulates Allergic Airway Inflammation in Mice.

    PubMed

    Pellaton, Céline; Nutten, Sophie; Thierry, Anne-Christine; Boudousquié, Caroline; Barbier, Nathalie; Blanchard, Carine; Corthésy, Blaise; Mercenier, Annick; Spertini, François

    2012-01-01

    Introduction. Preclinical and clinical evidences for a role of oral probiotics in the management of allergic diseases are emerging. Aim. We aimed at testing the immunomodulatory effects of intranasal versus intragastric administration of Lactobacillus paracasei NCC2461 in a mouse model of allergic airway inflammation and the specificity of different probiotics by comparing L. paracasei NCC2461 to Lactobacillus plantarum NCC1107. Methods. L. paracasei NCC2461 or L. plantarum NCC1107 strains were administered either intragastrically (NCC2461) or intranasally (NCC2461 or NCC1107) to OVA-sensitized mice challenged with OVA aerosols. Inflammatory cell recruitment into BALF, eotaxin and IL-5 production in the lungs were measured. Results. Intranasal L. paracasei NCC2461 efficiently protected sensitized mice upon exposure to OVA aerosols in a dose-dependent manner as compared to control mice. Inflammatory cell number, eotaxin and IL-5 were significantly reduced in BALF. Intranasal supplementation of L. paracasei NCC2461 was more potent than intragastric application in limiting the allergic response and possibly linked to an increase in T regulatory cells in the lungs. Finally, intranasal L. plantarum NCC1107 reduced total and eosinophilic lung inflammation, but increased neutrophilia and macrophages infiltration. Conclusion. A concerted selection of intervention schedule, doses, and administration routes (intranasal versus intragastric) may markedly contribute to modulate airway inflammation in a probiotic strain-specific manner. PMID:22762009

  2. Functional Analysis of the Fructooligosaccharide Utilization Operon in Lactobacillus paracasei 1195?

    PubMed Central

    Goh, Yong Jun; Lee, Jong-Hwa; Hutkins, Robert W.

    2007-01-01

    The fosABCDXE operon encodes components of a putative fructose/mannose phosphoenolpyruvate-dependent phosphotransferase system and a ?-fructosidase precursor (FosE) that are involved in the fructooligosaccharide (FOS) utilization pathway of Lactobacillus paracasei 1195. The presence of an N-terminal signal peptide sequence and an LPQAG cell wall anchor motif in the C-terminal region of the deduced FosE precursor amino acid sequence predicted that the enzyme is cell wall associated, indicating that FOS may be hydrolyzed extracellularly. In this study, cell fractionation experiments demonstrated that the FOS hydrolysis activity was present exclusively in the cell wall extract of L. paracasei previously grown on FOS. In contrast, no measurable FOS hydrolysis activity was detected in the cell wall extract from the isogenic fosE mutant. Induction of ?-fructosidase activity was observed when cells were grown on FOS, inulin, sucrose, or fructose but not when cells were grown on glucose. A diauxic growth pattern was observed when cells were grown on FOS in the presence of limiting glucose (0.1%). Analysis of the culture supernatant revealed that glucose was consumed first, followed by the longer-chain FOS species. Transcription analysis further showed that the fos operon was expressed only after glucose was depleted in the medium. Expression of fosE in a non-FOS-fermenting strain, Lactobacillus rhamnosus GG, enabled the recombinant strain to metabolize FOS, inulin, sucrose, and levan. PMID:17644636

  3. Probiotic Potential of Lactobacillus Strains with Antimicrobial Activity against Some Human Pathogenic Strains

    PubMed Central

    Shokryazdan, Parisa; Sieo, Chin Chin; Kalavathy, Ramasamy; Liang, Juan Boo; Alitheen, Noorjahan Banu; Faseleh Jahromi, Mohammad; Ho, Yin Wan

    2014-01-01

    The objective of this study was to isolate, identify, and characterize some lactic acid bacterial strains from human milk, infant feces, and fermented grapes and dates, as potential probiotics with antimicrobial activity against some human pathogenic strains. One hundred and forty bacterial strains were isolated and, after initial identification and a preliminary screening for acid and bile tolerance, nine of the best isolates were selected and further identified using 16?S rRNA gene sequences. The nine selected isolates were then characterized in vitro for their probiotic characteristics and their antimicrobial activities against some human pathogens. Results showed that all nine isolates belonged to the genus Lactobacillus. They were able to tolerate pH 3 for 3?h, 0.3% bile salts for 4?h, and 1.9?mg/mL pancreatic enzymes for 3?h. They exhibited good ability to attach to intestinal epithelial cells and were not resistant to the tested antibiotics. They also showed good antimicrobial activities against the tested pathogenic strains of humans, and most of them exhibited stronger antimicrobial activity than the reference strain L. casei Shirota. Thus, the nine Lactobacillus strains could be considered as potential antimicrobial probiotic strains against human pathogens and should be further studied for their human health benefits. PMID:25105147

  4. Generation of food-grade recombinant Lactobacillus casei delivering Myxococcus xanthus prolyl endopeptidase.

    PubMed

    Alvarez-Sieiro, Patricia; Martin, Maria Cruz; Redruello, Begoña; Del Rio, Beatriz; Ladero, Victor; Palanski, Brad A; Khosla, Chaitan; Fernandez, Maria; Alvarez, Miguel A

    2014-08-01

    Prolyl endopeptidases (PEP) (EC 3.4.21.26), a family of serine proteases with the ability to hydrolyze the peptide bond on the carboxyl side of an internal proline residue, are able to degrade immunotoxic peptides responsible for celiac disease (CD), such as a 33-residue gluten peptide (33-mer). Oral administration of PEP has been suggested as a potential therapeutic approach for CD, although delivery of the enzyme to the small intestine requires intrinsic gastric stability or advanced formulation technologies. We have engineered two food-grade Lactobacillus casei strains to deliver PEP in an in vitro model of small intestine environment. One strain secretes PEP into the extracellular medium, whereas the other retains PEP in the intracellular environment. The strain that secretes PEP into the extracellular medium is the most effective to degrade the 33-mer and is resistant to simulated gastrointestinal stress. Our results suggest that in the future, after more studies and clinical trials, an engineered food-grade Lactobacillus strain may be useful as a vector for in situ production of PEP in the upper small intestine of CD patients. PMID:24752841

  5. Plantaricin LD1: a bacteriocin produced by food isolate of Lactobacillus plantarum LD1.

    PubMed

    Gupta, Aabha; Tiwari, Santosh Kumar

    2014-04-01

    Plantaricin LD1, a bacteriocin produced by Lactobacillus plantarum LD1, was characterized for biochemical and antimicrobial properties. Bacteriocin showed stability at high temperatures (100 °C for 20 min and 121 °C for 15 min under 15 psi pressure), in a pH range of 2.0-8.0 and also in the presence of organic solvents, surfactants and detergents. The crude preparation was not affected by catalase, amylase and lipase but activity was reduced in the presence of pepsin, trypsin and proteinase K showing proteinaceous nature of the compound. The molecular weight of bacteriocin was found to be ?6.5 kDa, and antimicrobial activity was confirmed by bioassay. It inhibited not only related strains but also other Gram-positive and Gram-negative bacteria such as Lactobacillus curvatus NRRL B-4562, Lactococcus lactis subsp. lactis NRRL B-1821, Enterococcus faecium NRRL B-2354, Enterobacter cloacae NRRL B-14298, Micrococcus luteus, Staphylococcus aureus, urogenic Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi, Shigella flexneri and Vibrio sp. These properties of plantaricin LD1 suggest its applications not only in food safety but in therapeutics as well. PMID:24522411

  6. Characterization of ethanol fermentation waste and its application to lactic acid production by Lactobacillus paracasei.

    PubMed

    Moon, Se-Kwon; Lee, Julia; Song, Hyohak; Cho, Jung-Hee; Choi, Gi-Wook; Seung, Doyoung

    2013-05-01

    In this study, an ethanol fermentation waste (EFW) was characterized for use as an alternative to yeast extract for bulk fermentation processes. EFW generated from a commercial plant in which ethanol is produced from cassava/rice/wheat/barley starch mixtures using Saccharomyces cerevisiae was used for lactic acid production by Lactobacillus paracasei. The effects of temperature, pH, and duration on the autolysis of an ethanol fermentation broth (EFB) were also investigated. The distilled EFW (DEFW) contained significant amounts of soluble proteins (2.91 g/l), nitrogen (0.47 g/l), and amino acids (24.1 mg/l). The autolysis of the EFB under optimum conditions released twice as much amino acids than in the DEFW. Batch fermentation in the DEFW increased the final lactic acid concentration, overall lactic acid productivity, and lactic acid yield on glucose by 17, 41, and 14 %, respectively, in comparison with those from comparable fermentation in a lactobacillus growth medium (LGM) that contained 2 g/l yeast extract. Furthermore, the overall lactic acid productivity in the autolyzed then distilled EFW (ADEFW) was 80 and 27 % higher than in the LGM and DEFW, respectively. PMID:22907566

  7. Functional Analysis of the p40 and p75 Proteins from Lactobacillus casei BL23

    PubMed Central

    Bäuerl, Christine; Pérez-Martínez, Gaspar; Yan, Fang; Polk, D. Brent; Monedero, Vicente

    2011-01-01

    The genomes of Lactobacillus casei/paracasei and Lactobacillus rhamnosus strains carry two genes encoding homologues of p40 and p75 from L. rhamnosus GG, two secreted proteins which display anti-apoptotic and cell protective effects on human intestinal epithelial cells. p40 and p75 carry cysteine, histidine-dependent aminohydrolase/peptidase (CHAP) and NLPC/P60 domains, respectively, which are characteristic of proteins with cell-wall hydrolase activity. In L. casei BL23 both proteins were secreted to the growth medium and were also located at the bacterial cell surface. The genes coding for both proteins were inactivated in this strain. Inactivation of LCABL_00230 (encoding p40) did not result in a significant difference in phenotype, whereas a mutation in LCABL_02770 (encoding p75) produced cells that formed very long chains. Purified glutathione-S-transferase (GST)-p40 and -p75 fusion proteins were able to hydrolyze the muropeptides from L. casei cell walls. Both fusions bound to mucin, collagen and to intestinal epithelial cells and, similar to L. rhamnosus GG p40, stimulated epidermal growth factor receptor phosphorylation in mouse intestine ex vivo. These results indicate that extracellular proteins belonging to the machinery of cell-wall metabolism in the closely related L. casei/paracasei-L. rhamnosus group are most likely involved in the probiotic effects described for these bacteria PMID:21178363

  8. Detection and Identification of Probiotic Lactobacillus plantarum Strains by Multiplex PCR Using RAPD-Derived Primers

    PubMed Central

    Galanis, Alex; Kourkoutas, Yiannis; Tassou, Chrysoula C.; Chorianopoulos, Nikos

    2015-01-01

    Lactobacillus plantarum 2035 and Lactobacillus plantarum ACA-DC 2640 are two lactic acid bacteria (LAB) strains that have been isolated from Feta cheese. Both display significant potential for the production of novel probiotic food products. The aim of the present study was the development of an accurate and efficient method for the molecular detection and identification of the above strains in a single reaction. A multiplex PCR assay was designed for each strain, based on specific primers derived from Random Amplified Polymorphic DNA (RAPD) Sequenced Characterized Amplified Region (SCAR) analysis. The specificity of the assay was tested with a total of 23 different LAB strains, for L. plantarum 2035 and L. plantarum ACA-DC 2640. The multiplex PCR assay was also successfully applied for the detection of the above cultures in yogurt samples prepared in our lab. The proposed methodology may be applied for monitoring the presence of these strains in food products, thus evaluating their probiotic character. Moreover, our strategy may be adapted for other novel LAB strains with probiotic potential, thus providing a powerful tool for molecular discrimination that could be invaluable to the food industry. PMID:26506345

  9. Physicochemical and hygienic effects of Lactobacillus acidophilus in Iranian white cheese

    PubMed Central

    Mahmoudi, Razzaqh; Tajik, Hossein; Ehsani, Ali; Zare, Payman

    2012-01-01

    Increasing incidence of food-borne disease along with its social and economic consequences have led to conducting extensive research in order to produce safer food and develop new antimicrobial agents; among them, extensive use of probiotics and bacteriocins as biological additives is of significant importance. The aim of the present study was to evaluate the interactions (growth behavior and survival) of Listeria monocytogenes and Lactobacillus acidophilus in various stages of production, ripening and storage of Iranian white cheese. Changes in pH values at different stages of cheese ripening, along with changes in organoleptic properties of cheese were also assessed. Compared to other treatments, in the treatment of cheese with probiotic agent without starter, the most significant decrease in Listeria monocytogenes count at the end of ripening stage was observed (3.16 Log per gram cheese compared with the control group) (p < 0.05). Survival of probiotic bacteria in control samples of cheese were significantly higher when compared to cheese sample contaminated with Listeria (p < 0.05). White probiotic cheese with starter had the highest of sensory acceptability (p < 0.05). Listeria Monocytogenes count decreased during ripening period of probiotic white cheese but the bacteria survived in probiotic white cheese. Lactobacillus acidophilus count decreased during ripening period of white cheese but it did not lower to less than 106 CFU per g at the end of ripening and storage periods. PMID:25610568

  10. Lactobacillus casei strain Shirota protects against nonalcoholic steatohepatitis development in a rodent model.

    PubMed

    Okubo, Hirofumi; Sakoda, Hideyuki; Kushiyama, Akifumi; Fujishiro, Midori; Nakatsu, Yusuke; Fukushima, Toshiaki; Matsunaga, Yasuka; Kamata, Hideaki; Asahara, Takashi; Yoshida, Yasuto; Chonan, Osamu; Iwashita, Misaki; Nishimura, Fusanori; Asano, Tomoichiro

    2013-12-01

    Gut microbiota alterations are associated with various disorders. In this study, gut microbiota changes were investigated in a methionine-choline-deficient (MCD) diet-induced nonalcoholic steatohepatitis (NASH) rodent model, and the effects of administering Lactobacillus casei strain Shirota (LcS) on the development of NASH were also investigated. Mice were divided into three groups, given the normal chow diet (NCD), MCD diet, or the MCD diet plus daily oral administration of LcS for 6 wk. Gut microbiota analyses for the three groups revealed that lactic acid bacteria such as Bifidobacterium and Lactobacillus in feces were markedly reduced by the MCD diet. Interestingly, oral administration of LcS to MCD diet-fed mice increased not only the L. casei subgroup but also other lactic acid bacteria. Subsequently, NASH development was evaluated based on hepatic histochemical findings, serum parameters, and various mRNA and/or protein expression levels. LcS intervention markedly suppressed MCD-diet-induced NASH development, with reduced serum lipopolysaccharide concentrations, suppression of inflammation and fibrosis in the liver, and reduced colon inflammation. Therefore, reduced populations of lactic acid bacteria in the colon may be involved in the pathogenesis of MCD diet-induced NASH, suggesting normalization of gut microbiota to be effective for treating NASH. PMID:24113768

  11. Effect of a Selected Lactobacillus spp-Based Probiotic on Salmonella Serovar Enteritidis-Infected Broiler Chicks

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The effect of a Lactobacillus-based probiotic (FM-B11**TM) on Salmonella recovery was evaluated in liquid (Exp. 1) and lyophilized (Exp. 2) forms in two separate experiments with two trials each. For each trial, eighty broiler chicks were randomly allocated into two treatments: control and probioti...

  12. Draft Genome Sequence of Lactobacillus namurensis Chizuka 01, Isolated from Nukadoko, a Pickling Bed of Fermented Rice Bran.

    PubMed

    Kato, Keita; Toh, Hidehiro; Sakamoto, Naoshige; Mori, Kazuki; Tashiro, Kosuke; Hibi, Naruhiro; Sonomoto, Kenji; Nakayama, Jiro

    2014-01-01

    Lactobacillus namurensis Chizuka 01 was isolated from nukadoko, which is a fermented rice bran bed traditionally used in Japan for pickling vegetables. Here, we report the first draft of an annotated genome sequence of this organism. This paper is the first published report of the genomic sequence of L. namurensis. PMID:24504001

  13. Draft Genome Sequence of Lactobacillus namurensis Chizuka 01, Isolated from Nukadoko, a Pickling Bed of Fermented Rice Bran

    PubMed Central

    Kato, Keita; Toh, Hidehiro; Sakamoto, Naoshige; Mori, Kazuki; Tashiro, Kosuke; Hibi, Naruhiro; Sonomoto, Kenji

    2014-01-01

    Lactobacillus namurensis Chizuka 01 was isolated from nukadoko, which is a fermented rice bran bed traditionally used in Japan for pickling vegetables. Here, we report the first draft of an annotated genome sequence of this organism. This paper is the first published report of the genomic sequence of L. namurensis. PMID:24504001

  14. LACTOBACILLUS CASEI METABOLIC POTENTIAL TO UTILIZE CITRATE AS AN ENERGY SOURCE IN RIPENING CHEESE: A BIOINFORMATICS APPROACH

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This research describes a unique PCAC for Lactobacillus casei. Additionally, it describes the citric acid catabolism end-product by this non-starter lactic acid bacteria during growth, and under conditions similar to those present in ripening cheese. It provides insights on pathways preferably util...

  15. Complete genome sequence of Lactobacillus plantarum ZS2058, a probiotic strain with high conjugated linoleic acid production ability.

    PubMed

    Yang, Bo; Chen, Haiqin; Tian, Fengwei; Zhao, Jianxin; Gu, Zhennan; Zhang, Hao; Chen, Yong Q; Chen, Wei

    2015-11-20

    Lactobacillus plantarum ZS2058 was isolated from sauerkraut and identified to synthesize the beneficial metabolite conjugated linoleic acid. The genome contains a 319,7363-bp chromosome and three plasmids. The sequence will facilitate identification and characterization of the genetic determinants for its putative biological benefits. PMID:26439428

  16. Antimicrobial activity and partial characterization of bacteriocin-like inhibitory substances produced by Lactobacillus spp. isolated from artisanal Mexican cheese.

    PubMed

    Heredia-Castro, Priscilia Y; Méndez-Romero, José I; Hernández-Mendoza, Adrián; Acedo-Félix, Evelia; González-Córdova, Aarón F; Vallejo-Cordoba, Belinda

    2015-12-01

    Lactobacillus spp. from Mexican Cocido cheese were shown to produce bacteriocin-like substances (BLS) active against Staphylococcus aureus,Listeria innocua,Escherichia coli, andSalmonella typhimurium by using the disk diffusion method. Crude extracts of Lactobacillus fermentum showed strong inhibitory activity against Staph. aureus, L. innocua, E. coli, and Salmonella cholerae. Complete inactivation of antimicrobial activity was observed after treatment of crude extracts with proteinase K, pronase, papain, trypsin, and lysozyme, confirming their proteinaceous nature. However, antimicrobial activity was partly lost for some of the crude extracts when treated with ?-amylase, indicating that carbohydrate moieties were involved. The antimicrobial activity of the crude extracts was stable at 65°C for 30min over a wide pH range (2-8), and addition of potassium chloride, sodium citrate, ethanol, and butanol did not affect antibacterial activity. However, antimicrobial activity was lost after heating at 121°C for 15min, addition of methanol or Tween 80. Fourteen out of 18 Lactobacillus spp. showed antimicrobial activity against different test microorganisms, and 12 presented bacteriocin-like substances. Generation time and growth rate parameters indicated that the antimicrobial activity of crude extracts from 3 different strains was effective against the 4 indicator microorganisms. One of the crude extracts showed inhibition not only against gram-positive but also against gram-negative bacteria. Bacteriocin-like substances produced by this specific Lactobacillus strain showed potential for application as a food biopreservative. PMID:26476937

  17. Putative and unique gene sequence utilization for the design of species specific probes as modeled by Lactobacillus plantarum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The concept of utilizing putative and unique gene sequences for the design of species specific probes was tested. The abundance profile of assigned functions within the Lactobacillus plantarum genome was used for the identification of the putative and unique gene sequence, csh. The targeted gene (cs...

  18. Prevention of Mycobacterium avium subsp. paratuberculosis (MAP) Infection in BALB/c Mice by Feeding Probiotic Lactobacillus acidophilus NP-51

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this study was to examine effects of feeding Lactobacillus acidophilus strain NP51 to mice challenged with Mycobacterium avium subspecies paratuberculosis (MAP). Mice were randomized to ten treatment groups; sentinels, control, heat-killed MAP, viable MAP, heat-killed NP51, viable ...

  19. Prevention of Mycobacterium avium subsp. paratuberculosis Infection in BALB/c Mice by Feeding Lactobacillus acidophilus Strain NP-51

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The immune responses of 390 BALB/c mice fed the probiotic Lactobacillus acidophilus strain NP51® and infected with Mycobacterium avium subspecies paratuberculosis (MAP) were evaluated in a 6-month trial. Mice were randomized to nine treatment groups fed either viable- or heat-killed NP51 and inocula...

  20. Isolation of Lactobacillus salivarius 1077 (NRRL B-50053) and characterization of its bacteriocin and spectra of antimicrobial activity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Lactobacillus salivarius 1077 (NRRL B-50053) was isolated from poultry intestinal materials after demonstrating in-vitro anti-Campylobacter jejuni activity. The isolate was then used for in-vitro fermentation. The protein content of the cell-free supernatant from the spent medium was precipitated ...

  1. Metabolism of lactic acid in fermented cucumbers by Lactobacillus buchneri and related species, potential spoilage organisms in reduced salt fermentations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Recent evidence suggests that Lactobacillus buchneri may play an important role in spoilage-associated secondary fermentation of cucumbers. Lactic acid degradation during fermented cucumber spoilage is influenced by sodium chloride (NaCl) concentration, pH, and presence of oxygen. Objectives were to...

  2. Prevention of Mycobacterium avium subsp. paratuberculosis (MAP) Infection in Balb/c Mice by Feeding Probiotic Lactobacillus acidophilus NP-51

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this study was to examine effects of feeding Lactobacillus acidophilus strain NP51 to mice challenged with Mycobacterium avium subspecies paratuberculosis (MAP), the causative agent of Johne’s disease. We hypothesized that feeding NP51 would increase Th-1 responses and decrease prog...

  3. Prevention of Mycobacterium avium subsp. paratuberculosis (MAP) infection in BALB/c mice by feeding probiotic Lactobacillus acidophilus NP-51

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The objective of this study was to examine effects of feeding Lactobacillus acidophilus strain NP51 to mice challenged with Mycobacterium avium subspecies paratuberculosis (MAP). Mice were randomized to ten treatment groups; sentinels, control, heat-killed MAP, viable MAP, heat-killed NP51, viable ...

  4. Genome Sequence of Lactobacillus delbrueckii subsp. lactis CNRZ327, a Dairy Bacterium with Anti-Inflammatory Properties

    PubMed Central

    El Kafsi, Hela; Binesse, Johan; Loux, Valentin; Buratti, Julien; Boudebbouze, Samira; Dervyn, Rozenn; Hammani, Amal; Maguin, Emmanuelle

    2014-01-01

    Lactobacillus delbrueckii subsp. lactis CNRZ327 is a dairy bacterium with anti-inflammatory properties both in vitro and in vivo. Here, we report the genome sequence of this bacterium, which appears to contain no less than 215 insertion sequence (IS) elements, an exceptionally high number regarding the small genome size of the strain. PMID:25035318

  5. Genome Sequence of Lactobacillus delbrueckii subsp. lactis CNRZ327, a Dairy Bacterium with Anti-Inflammatory Properties.

    PubMed

    El Kafsi, Hela; Binesse, Johan; Loux, Valentin; Buratti, Julien; Boudebbouze, Samira; Dervyn, Rozenn; Hammani, Amal; Maguin, Emmanuelle; van de Guchte, Maarten

    2014-01-01

    Lactobacillus delbrueckii subsp. lactis CNRZ327 is a dairy bacterium with anti-inflammatory properties both in vitro and in vivo. Here, we report the genome sequence of this bacterium, which appears to contain no less than 215 insertion sequence (IS) elements, an exceptionally high number regarding the small genome size of the strain. PMID:25035318

  6. Heterophil Phagocytic Activity Stimulated by Lactobacillus salivarius L61 and L55 Supplementation in Broilers with Salmonella Infection

    PubMed Central

    Sornplang, Pairat; Leelavatcharamas, Vichai; Soikum, Chaiyaporn

    2015-01-01

    Newborn chicks are susceptible to Salmonella enterica serovar Enteritidis (SE). The objective of this study was to evaluate the effect of Lactobacillus probiotic isolated from chicken feces on heterophil phagocytosis in broiler chicks. A total of 150 newborn broiler chicks were divided into 5 groups (30 chicks per group) as follows: group 1 (normal control), given feed and water only, group 2 (positive control) given feed, water and SE infection, group 3 (L61 treated) given feed, water, SE infection followed by Lactobacillus salivarius L61 treatment, group 4 (L55 treated) given feed, water, SE infection followed by L. salivarius L55 treatment, and group 5 given feed, water, SE infection followed by L. salivarius L61 + L55 combination treatment. After SE infection, L. salivarius treatment lasted for 7 days. The results showed that L. salivarius L61 and L. salivarius L55 treatment, either alone or combination of both, increased the survival rate after SE infection, and upregulated heterophil phagocytosis and phagocytic index (PI). Conversely, chick groups treated with Lactobacillus showed lower SE recovery rate from cecal tonsils than that of the positive control group. The PI values of the chicken group with SE infection, followed by the combination of L. salivarius L61 and L. salivarius L55 were the highest as compared to either positive control or normal control group. Two Lactobacillus strains supplementation group showed significantly (p<0.05) higher PI value at 48 h than 24 h after treatment. PMID:26580288

  7. Gut-Resident Lactobacillus Abundance Associates with IDO1 Inhibition and Th17 Dynamics in SIV-Infected Macaques.

    PubMed

    Vujkovic-Cvijin, Ivan; Swainson, Louise A; Chu, Simon N; Ortiz, Alexandra M; Santee, Clark A; Petriello, Annalise; Dunham, Richard M; Fadrosh, Douglas W; Lin, Din L; Faruqi, Ali A; Huang, Yong; Apetrei, Cristian; Pandrea, Ivona; Hecht, Frederick M; Pilcher, Christopher D; Klatt, Nichole R; Brenchley, Jason M; Lynch, Susan V; McCune, Joseph M

    2015-11-24

    Gut microbes can profoundly modulate mucosal barrier-promoting Th17 cells in mammals. A salient feature of HIV/simian immunodeficiency virus (SIV) immunopathogenesis is the loss of Th17 cells, which has been linked to increased activity of the immunomodulatory enzyme, indoleamine 2,3-dioxygenase 1 (IDO 1). The role of gut microbes in this system remains unknown, and the SIV-infected rhesus macaque provides a well-described model for HIV-associated Th17 loss and mucosal immune disruption. We observed a specific depletion of gut-resident Lactobacillus during acute and chronic SIV infection of rhesus macaques, which was also seen in early HIV-infected humans. This depletion in rhesus macaques correlated with increased IDO1 activity and Th17 loss. Macaques supplemented with a Lactobacillus-containing probiotic exhibited decreased IDO1 activity during chronic SIV infection. We propose that Lactobacillus species inhibit mammalian IDO1 and thus may help to preserve Th17 cells during pathogenic SIV infection, providing support for Lactobacillus species as modulators of mucosal immune homeostasis. PMID:26586432

  8. Complete Genome Sequence of Potential Probiotic Lactobacillus sp. HFC8, Isolated from Human Gut Using PacBio SMRT Sequencing

    PubMed Central

    Kumari, Madhu; Swarnkar, Mohit Kumar; Kumar, Sanjay

    2015-01-01

    We report a 3.07-Mb complete genome sequence of a lactic acid bacterium, Lactobacillus sp. HFC8. The gene-coding clusters are predicated for probiotic characteristics, like bacteriocin production, cell adhesion, bile salt hydrolysis, lactose metabolism, autoaggregation, and tolerance to oxidative stress. PMID:26586884

  9. Draft Genome Sequence of Lactobacillus plantarum Strain SNU.Lp177 from Pig Feces in South Korea

    PubMed Central

    Jin, Gwi-Deuk; Lee, Jun-Yeong; Choi, Yun-Jaie

    2015-01-01

    Herein we report a draft genome sequence for Lactobacillus plantarum SNU.Lp177, which was isolated from animal gut pig feces in South Korea. The draft genome of L. plantarum SNU.Lp177 contains 3,204,772 bp with a G+C content of 44.98% in 101 contigs (N50 = 116,595 bp). PMID:26450744

  10. Reduced-fat Cheddar and Swiss-type cheeses harboring exopolysaccharide-producing probiotic Lactobacillus mucosae DPC 6426.

    PubMed

    Ryan, P M; Burdíková, Z; Beresford, T; Auty, M A E; Fitzgerald, G F; Ross, R P; Sheehan, J J; Stanton, C

    2015-12-01

    Exopolysaccharide-producing Lactobacillus mucosae DPC 6426 was previously shown to have promising hypocholesterolemic activity in the atherosclerosis-prone apolipoprotein-E-deficient (apoE(-/-)) murine model. The aim of this study was to investigate the suitability of reduced-fat Cheddar and Swiss-type cheeses as functional (carrier) foods for delivery of this probiotic strain. All cheeses were manufactured at pilot-scale (500-L vats) in triplicate, with standard commercially available starters: for Cheddar, Lactococcus lactis; and for Swiss-type cheese, Streptococcus thermophilus, Lactobacillus helveticus, and Propionibacterium freudenreichii. Lactobacillus mucosae DPC 6426 was used as an adjunct culture during cheese manufacture, at a level of ~10(6) cfu·mL(-1) cheese milk (subsequently present in the cheese curd at>10(7) cfu·g(-1)). The adjunct strain remained viable at >5×10(7) cfu·g(-1) in both Swiss-type and Cheddar cheeses following ripening for 6 mo. Sensory analysis revealed that the presence of the adjunct culture imparted a more appealing appearance in Swiss-type cheese, but had no significant effect on the sensory characteristics of Cheddar cheeses. Moreover, the adjunct culture had no significant effect on cheese composition, proteolysis, pH, or instrumentally quantified textural characteristics of Cheddar cheeses. These data indicate that low-fat Swiss-type and Cheddar cheeses represent suitable food matrices for the delivery of the hypocholesterolemic Lactobacillus mucosae DPC 6426 in an industrial setting. PMID:26409971

  11. High quality draft genome of Lactobacillus kunkeei EFB6, isolated from a German European foulbrood outbreak of honeybees

    PubMed Central

    2015-01-01

    The lactic acid bacterium Lactobacillus kunkeei has been described as an inhabitant of fructose-rich niches. Here we report on the genome sequence of L. kunkeei EFB6, which has been isolated from a honeybee larva infected with European foulbrood. The draft genome comprises 1,566,851 bp and 1,417 predicted protein-encoding genes. PMID:26203329

  12. Enzyme Activities Affecting End Product Distribution by Lactobacillus plantarum in Response to Changes in pH and O2†

    PubMed Central

    Tseng, Ching-Ping; Montville, Thomas J.

    1990-01-01

    Lactobacillus plantarum catabolic end products changed in response to environmental conditions. While lactate was always the major end product, acetate was produced in alkaline and aerobic environments. Acetoin levels decreased under alkaline conditions. Changes in acetoin dehydrogenase, acetate kinase, NADH oxidase, pyruvate oxidase, and acetate kinase activities correlated with changes in end product distribution. PMID:16348283

  13. The Effects of Lactobacillus acidophilus on the Intestinal Smooth Muscle Contraction through PKC/MLCK/MLC Signaling Pathway in TBI Mouse Model

    PubMed Central

    Fang, Huan; Zhu, Lina; Gao, Ning; Zhu, Jingci

    2015-01-01

    Clinical studies have shown that probiotics influence gastrointestinal motility. However, the molecular mechanisms by which probiotic Lactobacillus modulates intestinal motility in traumatic brain injury (TBI) mouse model have not been explored. In the present study, we provided evidence showing that treatment of TBI mice with Lactobacillus acidophilus significantly improved the terminal ileum villus morphology, restored the impaired interstitial cells of Cajal (ICC) and the disrupted ICC networks after TBI, and prevented TBI-mediated inhibition of contractile activity in intestinal smooth muscle. Mechanistically, the decreased concentration of MLCK, phospho-MLC20 and phospho-MYPT1 and increased concentration of MLCP and PKC were observed after TBI, and these events mediated by TBI were efficiently prevented by Lactobacillus acidophilus application. These findings may provide a novel mechanistic basis for the application of Lactobacillus acidophilus in the treatment of TBI. PMID:26030918

  14. Surface displaced alfa-enolase of Lactobacillus plantarum is a fibronectin binding protein

    PubMed Central

    Castaldo, Cristiana; Vastano, Valeria; Siciliano, Rosa Anna; Candela, Marco; Vici, Manuela; Muscariello, Lidia; Marasco, Rosangela; Sacco, Margherita

    2009-01-01

    Background Lactic acid bacteria of the genus Lactobacillus and Bifidobacterium are one of the most important health promoting groups of the human intestinal microbiota. Their protective role within the gut consists in out competing invading pathogens for ecological niches and metabolic substrates. Among the features necessary to provide health benefits, commensal microorganisms must have the ability to adhere to human intestinal cells and consequently to colonize the gut. Studies on mechanisms mediating adhesion of lactobacilli to human intestinal cells showed that factors involved in the interaction vary mostly among different species and strains, mainly regarding interaction between bacterial adhesins and extracellular matrix or mucus proteins. We have investigated the adhesive properties of Lactobacillus plantarum, a member of the human microbiota of healthy individuals. Results We show the identification of a Lactobacillus plantarum LM3 cell surface protein (48 kDa), which specifically binds to human fibronectin (Fn), an extracellular matrix protein. By means of mass spectrometric analysis this protein was identified as the product of the L. plantarum enoA1 gene, coding the EnoA1 alfa-enolase. Surface localization of EnoA1 was proved by immune electron microscopy. In the mutant strain LM3-CC1, carrying the enoA1 null mutation, the 48 kDa adhesin was not anymore detectable neither by anti-enolase Western blot nor by Fn-overlay immunoblotting assay. Moreover, by an adhesion assay we show that LM3-CC1 cells bind to fibronectin-coated surfaces less efficiently than wild type cells, thus demonstrating the significance of the surface displaced EnoA1 protein for the L. plantarum LM3 adhesion to fibronectin. Conclusion Adhesion to host tissues represents a crucial early step in the colonization process of either pathogens or commensal bacteria. We demonstrated the involvement of the L. plantarum Eno A1 alfa-enolase in Fn-binding, by studying LM3 and LM3-CC1 surface proteins. Isolation of LM3-CC1 strain was possible for the presence of expressed enoA2 gene in the L. plantarum genome, giving the possibility, for the first time to our knowledge, to quantitatively compare adhesion of wild type and mutant strain, and to assess doubtless the role of L. plantarum Eno A1 as a fibronectin binding protein. PMID:19220903

  15. Probiotics Lactobacillus rhamnosus GG, Lactobacillus acidophilus suppresses DMH-induced procarcinogenic fecal enzymes and preneoplastic aberrant crypt foci in early colon carcinogenesis in Sprague Dawley rats.

    PubMed

    Verma, Angela; Shukla, Geeta

    2013-01-01

    Diet makes an important contribution to colorectal cancer (CRC) risk implying risks for CRC are potentially reducible. Therefore, the probiotics have been suggested as the prophylactic measure in colon cancer. In this study, different probiotics were used to compare their protective potential against 1,2 dimethylhydrazine dihydrochloride (DMH)-induced chemical colon carcinogenesis in Sprague Dawley rats. Animals belonging to different probiotic groups were fed orally with 1 × 10(9) lactobacilli daily for 1 week, and then a weekly injection of DMH was given intraperitoneally for 6 wks with daily administration of probiotic. Lactobacillus GG and L.acidophilus + DMH-treated animals had maximum percent reduction in ACF counts. A significant decrease (P < 0.05) in fecal nitroreductase activity was observed in L.casei + DMH and L.plantarum + DMH-treated rats whereas ?-glucuronidase activity decreased in L.GG + DMH and L.acidophilus + DMH-treated rats. Animals treated with Bifidobacterium bifidum + DMH had significant decreased ?-glucosidase activity. However, not much difference was observed in the colon morphology of animals belonging to various probiotic + DMH-treated rats compared with DMH-treated alone. The results indicated that probiotics, L.GG, and L.acidophilus can be used as the better prophylactic agents for experimental colon carcinogenesis. PMID:23368917

  16. Staphylococcus aureus toxic shock syndrome toxin-1 (TSST-1) production and Lactobacillus species growth in a defined medium simulating vaginal secretions.

    PubMed

    Stingley, Robin L; Liu, Huanli; Mullis, Lisa B; Elkins, Christopher A; Hart, Mark E

    2014-11-01

    Lactobacillus species are commensal with the healthy vaginal environment and inhibit the growth of many pathogenic bacteria in the vaginal tract by a variety of mechanisms, such as the production of hydrogen peroxide, organic acids, and antimicrobial substances. Simulation of the vaginal environment is crucial for proper investigation of the effects of Lactobacillus species on pathogenic bacteria. In this study, we modified a medium used to simulate vaginal secretions to improve the growth of toxic shock syndrome toxin-1 (TSST-1)-producing Staphylococcus aureus clinical strains and Lactobacillus species so that interactions between these bacteria may be examined. A medium consisting of basal salts, vitamins, albumin, glycogen, mucin, urea, sodium bicarbonate, polyoxyethylene sorbitan monolaurate, and amino acids supported the growth of S. aureus and the production of TSST-1 as determined by Western analysis. Improved growth of the Lactobacillus species was seen when this same medium was supplemented with manganese chloride, sodium acetate, and an increase in glucose concentration. However, growth of S. aureus in the supplemented medium resulted in reduced levels of TSST-1. Production of TSST-1 was not detected in a medium routinely used for the growth of Lactobacillus species although S. aureus growth was not inhibited. The development of an improved genital tract secretion medium provides a more authentic environment in which to study the interactions of Lactobacillus species and vaginal pathogens, such as S. aureus. PMID:25135489

  17. Effects of 5-hydroxytryptophan and m-hydroxybenzylhydrazine associated to Lactobacillus spp. on the humoral response of broilers challenged with Salmonella Enteritidis.

    PubMed

    Donato, T C; Baptista, A A S; Garcia, K C O D; Smaniotto, B D; Okamoto, A S; Sequeira, J L; Andreatti Filho, R L

    2015-09-01

    This study investigates the effects of different doses of serotonin, its precursor 5-hydroxytry-ptophan (5HTP), and m-hydroxybenzylhydrazine inhibitor (NSD1015), administered via intraperitoneal for 5 consecutive days, on behavior and average body weight of broilers. We also measured the humoral immune response and quantification of Salmonella Enteritidis in broilers chickens that received the drugs evaluated and a Lactobacillus pool. The study was divided into 3 experiments: Experiment 1--administration of pharmaceuticals with choice of dosage; Experiment 2--administration of pharmaceuticals and a Lactobacillus pool in birds that were not challenged with S. Enteritidis, and Experiment 3--administration of pharmaceuticals and a Lactobacillus pool in birds challenged with S. Enteritidis. The ELISA was used to scan dosages of intestinal IgA and serum IgY. We used colony-forming units to quantify S. Enteritidis. The concentrations of IgA and IgY did not show significant differences (P>0.05) in Experiment 2. In Experiment 3, NSD1015 associated with Lactobacillus determined higher IgA concentrations, promoting greater stimulus to the immune system than 5HTP. Regarding quantification of S. Enteritidis in the cecal content of birds, 5HTP associated to Lactobacillus determined the smallest number of bacteria, showing possible interaction of 5-hydroxytryptophan and Lactobacillus spp. with the immune system of broiler chickens. PMID:26195810

  18. Pre-alcoholic fermentation acidification of red grape must using Lactobacillus plantarum.

    PubMed

    Onetto, Cristóbal A; Bordeu, Edmundo

    2015-12-01

    Red grape musts from overripe grapes are characterised by high pH and sugar concentration. Corrections with organic acids are commonly used to secure the alcoholic fermentation and improve the organoleptic characteristics of the wine. In this study we test an alternative biological acidification method using the ability of Lactobacillus plantarum to produce high concentrations of lactic acid. The time course of sugars, organic acids and pH were measured. Available sugars were consumed by L. plantarum producing up to 8.3 g L(-1) of lactic acid. Lactic acid changed the pH from 3.9 to 3.4 after 14 days post-inoculation without yielding a relevant concentration of acetic acid (0.34 g L(-1)). PMID:26437637

  19. [Study of probiotic properties of bacteriocinogenic Lactobacillus plantarum G3/3(13) strain].

    PubMed

    Dimova, M I

    2006-01-01

    Bacteriocinogenic culture Lactobacillus plantarum G3/3(13) was tested according to the basic requirements to probiotic strains. The reidentification of the strain was carried out using a set of methods including PCR. It was confirmed that the strain belongs to the species L. plantarum. The studied culture expressed antagonistic activity with respect to Enterococcus faecalis and E. faecium. The susceptibility test to 57 antibiotics of different groups showed that the strain was resistant to oxacillin, cefepyme, lomefloxacin, polymyxin, nalidixic acid and the antifungal antibiotics. High adhesive properties of the microorganism to the human epithelial cells (medium adhesion index was 4.6-5.6; microorganism adhesion index was 4.9-5.9) were shown. Obtained results allow to recommend L. plantarum G3/3(13) for the construction of probiotics. PMID:17100328

  20. Isolation and typification of histamine-producing Lactobacillus vaginalis strains from cheese.

    PubMed

    Diaz, Maria; Del Rio, Beatriz; Ladero, Victor; Redruello, Begoña; Fernández, María; Martin, Maria Cruz; Alvarez, Miguel A

    2015-12-23

    In food, the biogenic amine (BA) histamine is mainly produced by histidine decarboxylation catalysed by microbial histidine decarboxylase. The consumption of foods containing high concentrations of histamine can trigger adverse neurological, gastrointestinal and respiratory reactions. Indeed, histamine is one of the most toxic of all BAs, and is often detected in high concentration in cheese. However, little is known about the microorganisms responsible for its accumulation in this food. In the present work, 25 histamine-producing Lactobacillus vaginalis strains were isolated from a blue-veined cheese (the first time that histamine-producing strains of this species have been isolated from any food). The restriction profiles of their genomes were analysed by PFGE, and seven lineages identified. The presence of the histidine decarboxylase gene (hdcA) was confirmed by PCR. The nucleotide sequence and genetic organisation of the histamine biosynthesis gene cluster (HDC) and its flanking regions are described for a representative strain (L. vaginalis IPLA11050). PMID:26394683

  1. Lactic acid production from sugar-cane juice by a newly isolated Lactobacillus sp.

    PubMed

    Timbuntam, Walaiporn; Sriroth, Klanarong; Tokiwa, Yutaka

    2006-06-01

    A newly isolated sucrose-tolerant, lactic acid bacterium, Lactobacillus sp. strain FCP2, was grown on sugar-cane juice (125 g sucrose l(-1), 8 g glucose l(-1) and 6 g fructose l(-1)) for 5 days and produced 104 g lactic acid l(-1) with 90% yield. A higher yield (96%) and productivity (2.8 g l(-1 )h(-1)) were obtained when strain FCP2 was cultured on 3% w/v (25 g sucrose l(-1), 2 g glucose l(-1) and 1 g fructose l(-1)) sugar-cane juice for 10 h. Various cheap nitrogen sources such as silk worm larvae, beer yeast autolysate and shrimp wastes were also used as a substitute to yeast extract. PMID:16786246

  2. Synthesis of oligo- and polysaccharides by Lactobacillus reuteri 121 reuteransucrase at high concentrations of sucrose.

    PubMed

    Meng, Xiangfeng; Dobruchowska, Justyna M; Gerwig, Gerrit J; Kamerling, Johannis P; Dijkhuizen, Lubbert

    2015-09-23

    GTFA, a glucansucrase enzyme of the probiotic bacterium Lactobacillus reuteri 121, is capable of synthesizing an ?-glucan polysaccharide with (1?4) and (1?6) linkages from sucrose. With respect to its biosynthesis, the present study has shown that the ratio of oligosaccharide versus polysaccharide synthesized was directly proportional to the concentration of sucrose. It appears that the size distribution of products is kinetically controlled, but the linkage distribution in the polysaccharide material is not changed. At high sucrose concentrations the sucrose isomers leucrose and trehalulose were synthesized, using the accumulated fructose as acceptor, together with 4'- and 6'-?-d-glucosyl-leucrose and 6'-?-d-glucosyl-trehalulose. The finding of an additional branched hexasaccharide demonstrates that the enzyme is able to introduce branch-points already in relatively short oligosaccharides. PMID:26281004

  3. Probiotic Characteristics of Lactobacillus plantarum FH185 Isolated from Human Feces

    PubMed Central

    2015-01-01

    Lactobacillus plantarum FH185 was isolated from the feces of healthy adults. In our previous study, L. plantarum FH185 was demonstrated that it has anti-obesity effect in the in vitro and in vivo test. In order to determine its potential for use as a probiotic, we investigated the physiological characteristics of L. plantarum FH185. The optimum growth temperature of L. plantarum FH185 was 40?. L. plantarum FH185 showed higher sensitivity to novobiocin in a comparison of fifteen different antibiotics and showed higher resistance to polymyxin B and vancomycin. It also showed higher ?-galactosidase and N-acetyl-?-glucosaminidase activities. Moreover, it was comparatively tolerant to bile juice and acid, and inhibited the growths of Salmonella Typhimurium and Staphylococcus aureus with rates of 44.76% and 53.88%, respectively. It also showed high adhesion activity to HT-29 cells compared to L. rhamnosus GG.

  4. Statistical optimization of alpha-amylase production by probiotic Lactobacillus plantarum MTCC 1407 in submerged fermentation.

    PubMed

    Panda, Smita H; Swain, Manas R; Kar, Shaktimay; Ray, Ramesh C; Montet, Dider

    2008-01-01

    Production and purification of alpha-amylase by probiotic Lactobacillus plantarum MTCC 1407 has been investigated under submerged fermentation using Mann Rogassa Sharpe medium containing (1%) soluble starch in lieu of glucose (2%) as carbon source. Response Surface Methodology was used to evaluate the effect of main variables, i.e. incubation period, pH and temperature on enzyme production. A full factorial Central Composite Design was applied to study these main factors that affected alpha-amylase production. The experimental results showed that the optimum incubation period, pH and temperature were 36 h, 7.0 and 35 degrees C, respectively. The purified enzyme (by ammonium sulphate precipitation) had a molecular mass of 75 450 Da in SDS-PAGE. PMID:18646403

  5. Purification of collagen-binding proteins of Lactobacillus reuteri NCIB 11951.

    PubMed

    Aleljung, P; Shen, W; Rozalska, B; Hellman, U; Ljungh, A; Wadström, T

    1994-04-01

    Collagen type-I-binding proteins of Lactobacillus reuteri NCIB 11951 were purified. The cell surface proteins were affinity purified on collagen Sepharose and eluted with an NaCl gradient. Two protein bands were eluted from the column (29 kDa and 31 kDa), and both bound radio-labeled collagen type I. Rabbit antisera raised against the 29 kDa and 31 kDa protein reacted with the affinity-purified proteins in a Western blot with whole-cell extract used as antigen. The N-terminal sequence of the 29-kDa and 31-kDa proteins demonstrated the closest homologies with internal sequences from an Escherichia coli trigger factor protein (TIG.ECOLI). Out of nine other lactobacilli, the antisera reacted only with the L. reuteri and not with the other species tested. PMID:7764651

  6. Role of probiotic Lactobacillus fermentum KKL1 in the preparation of a rice based fermented beverage.

    PubMed

    Ghosh, Kuntal; Ray, Mousumi; Adak, Atanu; Halder, Suman K; Das, Arpan; Jana, Arijit; Parua Mondal, Saswati; Vágvölgyi, Csaba; Das Mohapatra, Pradeep K; Pati, Bikas R; Mondal, Keshab C

    2015-07-01

    A dominant lactic acid bacteria, Lactobacillus fermentum KKL1 was isolated from an Indian rice based fermented beverage and its fermentative behavior on rice was evaluated. The isolate grown well in rice and decreased the pH, with an increase of total titratable acidity on account of high yield in lactic acid and acetic acid. The production of ?-amylase and glucoamylase by the strain reached plateau on 1st and 2nd day of fermentation respectively. The accumulation of malto-oligosaccharides of different degrees of polymerization was also found highest on 4th day. Besides, phytase activity along with accumulation of free minerals also unremittingly increased throughout the fermentation. The fermented materials showed free radical scavenging activity against DPPH radicals. In-vitro characteristics revealed the suitability of the isolate as probiotic organism. The above profiling revealed that probiotic L. fermentum KKL1 have the significant impact in preparation of rice beer and improves its functional characteristics. PMID:25693452

  7. Biotransformation of caffeoyl quinic acids from green coffee extracts by Lactobacillus johnsonii NCC 533

    PubMed Central

    2013-01-01

    The potential of Lactobacillus johnsonii NCC 533 to metabolize chlorogenic acids from green coffee extract was investigated. Two enzymes, an esterase and a hydroxycinnamate decarboxylase (HCD), were involved in this biotransformation. The complete hydrolysis of 5-caffeoylquinic acid (5-CQA) into caffeic acid (CA) by L. johnsonii esterase occurred during the first 16 h of reaction time. No dihydrocaffeic acid was identified in the reaction mixture. The decarboxylation of CA into 4-vinylcatechol (4-VC) started only when the maximum concentration of CA was reached (10 ?mol/ml). CA was completely transformed into 4-VC after 48 h of incubation. No 4-vinylphenol or other derivatives could be identified in the reaction media. In this study we demonstrate the capability of L. johnsonii to transform chlorogenic acids from green coffee extract into 4-VC in two steps one pot reaction. Thus, the enzymatic potential of certain lactobacilli might be explored to generate flavor compounds from plant polyphenols. PMID:23692950

  8. Biotransformation of caffeoyl quinic acids from green coffee extracts by Lactobacillus johnsonii NCC 533.

    PubMed

    Bel-Rhlid, Rachid; Thapa, Dinesh; Kraehenbuehl, Karin; Hansen, Carl Erik; Fischer, Lutz

    2013-01-01

    The potential of Lactobacillus johnsonii NCC 533 to metabolize chlorogenic acids from green coffee extract was investigated. Two enzymes, an esterase and a hydroxycinnamate decarboxylase (HCD), were involved in this biotransformation. The complete hydrolysis of 5-caffeoylquinic acid (5-CQA) into caffeic acid (CA) by L. johnsonii esterase occurred during the first 16 h of reaction time. No dihydrocaffeic acid was identified in the reaction mixture. The decarboxylation of CA into 4-vinylcatechol (4-VC) started only when the maximum concentration of CA was reached (10 ?mol/ml). CA was completely transformed into 4-VC after 48 h of incubation. No 4-vinylphenol or other derivatives could be identified in the reaction media. In this study we demonstrate the capability of L. johnsonii to transform chlorogenic acids from green coffee extract into 4-VC in two steps one pot reaction. Thus, the enzymatic potential of certain lactobacilli might be explored to generate flavor compounds from plant polyphenols. PMID:23692950

  9. Cofactor-induced reversible folding of Flavodoxin-4 from Lactobacillus acidophilus.

    PubMed

    Dutta, Samit Kumar; Serrano, Pedro; Geralt, Michael; Axelrod, Herbert L; Xu, Qingping; Lesley, Scott A; Godzik, Adam; Deacon, Ashley M; Elsliger, Marc-André; Wilson, Ian A; Wüthrich, Kurt

    2015-10-01

    Flavodoxins in combination with the flavin mononucleotide (FMN) cofactor play important roles for electron transport in prokaryotes. Here, novel insights into the FMN-binding mechanism to flavodoxins-4 were obtained from the NMR structures of the apo-protein from Lactobacillus acidophilus (YP_193882.1) and comparison of its complex with FMN. Extensive reversible conformational changes were observed upon FMN binding and release. The NMR structure of the FMN complex is in agreement with the crystal structure (PDB ID: 3EDO) and exhibits the characteristic flavodoxin fold, with a central five-stranded parallel ?-sheet and five ?-helices forming an ?/?-sandwich architecture. The structure differs from other flavoproteins in that helix ?2 is oriented perpendicular to the ?-sheet and covers the FMN-binding site. This helix reversibly unfolds upon removal of the FMN ligand, which represents a unique structural rearrangement among flavodoxins. PMID:26177955

  10. Availability of Essential B-Group Vitamins to Lactobacillus plantarum in Green Olive Fermentation Brines

    PubMed Central

    Ruiz-Barba, J. L.; Jimenez-Diaz, R.

    1995-01-01

    The availability throughout the traditional Spanish-style green olive fermentation of four vitamins that are essential for the growth of Lactobacillus plantarum was studied. It was found that nicotinic and pantothenic acids, biotin, and vitamin B(inf6) were available in the fermentation brines within the first few days of the process, and their levels throughout the fermentative process were well above those required by L. plantarum to grow at its maximum growth rate. In laboratory medium, various yeast strains isolated from the fermentations were found to produce these vitamins in amounts several times that required by L. plantarum. This finding suggests that some yeast species might play a role in encouraging the growth of L. plantarum in Spanish-style green olive fermentation. PMID:16534988

  11. Characterization of Oaxaca raw milk cheese microbiota with particular interest in Lactobacillus strains.

    PubMed

    Caro, Irma; Mateo, Javier; Sandoval, María H; Soto, Sergio; García-Armesto, María R; Castro, José M

    2013-06-01

    The aim of this work was to identify and characterize lactobacilli strains from Mexican Oaxaca cheese. Twenty-seven lactobacilli isolated from Oaxaca cheese were identified at species level by 16S rRNA sequencing. Selected isolates were further characterized by ribotyping. Isolates were screened, among others, by acidifying capacity, antibiotic resistance, and activity against pathogens. Lactobacillus plantarum was predominant in Oaxaca cheese. The intraspecies variability of Lb. plantarum isolates was great. Multiple antibiotic resistances were observed. Eight isolates showed antimicrobial activity against the pathogenic species tested. Four Lb. plantarum strains showing low antibiotic resistance index, antimicrobial activity against enterotoxigenic Staphylococcus aureus and Listeria innocua stains, amine-negative decarboxylase activity, and resistance to NaCl and bile salt solutions, could be preselected to complete studies focused on designing a culture for use in pasteurized-milk Oaxaca cheese manufacturing. PMID:23548294

  12. Cancer Preventive Potential of Kimchi Lactic Acid Bacteria (Weissella cibaria, Lactobacillus plantarum)

    PubMed Central

    Kwak, Shin-Hye; Cho, Young-Mi; Noh, Geon-Min; Om, Ae-Son

    2014-01-01

    The number of death due to cancer has been increasing in Korea. Chemotherapy is known to cause side effects because it damages not only cancerous cells but healthy cells. Recently, attention has focused on food-derived chemopreventive and anti-tumor agents or formulations with fewer side effects. Kimchi, most popular and widely consumed in Korea, contains high levels of lactic acid bacteria and has been shown to possess chemopreventive effects. This review focuses on Weissella cibaria and Lactobacillus plantarum, the representatives of kimchi lactic acid bacteria, in terms of their abilities to prevent cancer. Further studies are needed to understand the mechanisms by which lactic acid bacteria in kimchi prevent carcinogenic processes and improve immune functions. PMID:25574459

  13. Enantioselective reduction of pentoxifylline to lisofylline using whole-cell Lactobacillus kefiri biotransformation.

    PubMed

    Pekala, Elzbieta; Godawska-Matysik, Anna; Zelaszczyk, Dorota

    2007-04-01

    Lisofylline (LSF) is a drug candidate that has been under investigation for acute respiratory distress syndrome, acute lung injury, septic shock and mucositis. As LSF is not commercially available in our country, we produced it for pharmacokinetic studies. In the present work whole-cell reduction of pentoxifylline [1-(5-oxohexyl)-3,5-dimethylxanthine] to LSF [1-(5R-hydroxyhexyl)-3,5-dimethylxanthine] using Lactobacillus kefiri DSM 20587 was investigated. Glucose or 2-propanol was used as a co-substrate to regenerate the NADPH cofactor. The reaction conditions were optimized. The influence of different concentrations of co-substrates on the yield and enantioselectivity of the biotransformation of pentoxifylline into LSF were tested. Maximum yield (100%) of biotransformation was reached in the presence of glucose as a co-substrate. At glucose concentrations of 675 and 900 mM the bioreduction of pentoxifylline proceeded highly enantioselectively (enantiomeric excess for the R enantiomer of 98%). PMID:17285680

  14. Lactobacillus acidophilus CRL 1014 improved “gut health” in the SHIME® reactor

    PubMed Central

    2013-01-01

    Background How to maintain “gut health” is a goal for scientists throughout the world. Therefore, microbiota management models for testing probiotics, prebiotics, and synbiotics have been developed. Methods The SHIME® model was used to study the effect of Lactobacillus acidophilus 1014 on the fermentation pattern of the colon microbiota. Initially, an inoculum prepared from human feces was introduced into the reactor vessels and stabilized over 2-wk using a culture medium. This stabilization period was followed by a 2-wk control period during which the microbiota was monitored. The microbiota was then subjected to a 4-wk treatment period by adding 5 mL of sterile peptone water with L. acidophilus CRL1014 at the concentration of 108 CFU/mL to vessel one (the stomach compartment). Plate counts, Denaturing Gradient Gel Electrophoresis (DGGE), short-chain fatty acid (SCFA) and ammonium analyses were carried out for monitoring of the microbial community from the colon compartments. Results A significant increase (p?Lactobacillus spp. and Bifidobacterium spp. populations was observed during the treatment period. The DGGE obtained showed changes in the lactobacilli community from the colon compartments of the SHIME® reactor. The (SCFA) concentration increased (p?

  15. Biochemical characterization of the major N-acetylmuramidase from Lactobacillus buchneri

    PubMed Central

    Anzengruber, Julia; Courtin, Pascal; Claes, Ingmar J. J.; Debreczeny, Monika; Hofbauer, Stefan; Obinger, Christian; Chapot-Chartier, Marie-Pierre; Vanderleyden, Jos; Messner, Paul; Schäffer, Christina

    2015-01-01

    Bacterial cell wall hydrolases are essential for peptidoglycan remodelling in regard to bacterial cell growth and division. In this study, peptidoglycan hydrolases (PGHs) of different Lactobacillus buchneri strains were investigated. First, the genome sequence of L. buchneri CD034 and L. buchneri NRRL B-30929 was analysed in silico for the presence of PGHs. Of 23 putative PGHs with different predicted hydrolytic specificities, the glycosyl hydrolase family 25 domain-containing homologues LbGH25B and LbGH25N from L. buchneri CD034 and NRRL B-30929, respectively, were selected and characterized in detail. Zymogram analysis confirmed hydrolysing activity on bacterial cell walls for both enzymes. Subsequent reversed-phase HPLC and MALDI-TOF MS analysis of the peptidoglycan breakdown products from L. buchneri strains CD034 and NRRL B-30929, and from Lactobacillus rhamnosus GG, which served as a reference, revealed that LbGH25B and LbGH25N have N-acetylmuramidase activity. Both enzymes were identified as cell wall-associated proteins by means of immunofluorescence microscopy and cellular fractionation, as well as by the ability of purified recombinant LbGH25B and LbGH25N to bind to L. buchneri cell walls in vitro. Moreover, similar secondary structures mainly composed of ?-sheets and nearly identical thermal stabilities with Tm values around 49 6C were found for the two N-acetylmuramidases by far-UV circular dichroism spectroscopy. The functional and structural data obtained are discussed and compared to related PGHs. In this study, a major N-acetylmuramidase from L. buchneri was characterized in detail for the first time. PMID:24858286

  16. Identification, Purification, and Characterization of a Novel Amino Acid Racemase, Isoleucine 2-Epimerase, from Lactobacillus Species

    PubMed Central

    Mutaguchi, Yuta; Ohmori, Taketo; Wakamatsu, Taisuke; Doi, Katsumi

    2013-01-01

    Accumulation of d-leucine, d-allo-isoleucine, and d-valine was observed in the growth medium of a lactic acid bacterium, Lactobacillus otakiensis JCM 15040, and the racemase responsible was purified from the cells and identified. The N-terminal amino acid sequence of the purified enzyme was GKLDKASKLI, which is consistent with that of a putative ?-aminobutyrate aminotransferase from Lactobacillus buchneri. The putative ?-aminobutyrate aminotransferase gene from L. buchneri JCM 1115 was expressed in recombinant Escherichia coli and then purified to homogeneity. The enzyme catalyzed the racemization of a broad spectrum of nonpolar amino acids. In particular, it catalyzed at high rates the epimerization of l-isoleucine to d-allo-isoleucine and d-allo-isoleucine to l-isoleucine. In contrast, the enzyme showed no ?-aminobutyrate aminotransferase activity. The relative molecular masses of the subunit and native enzyme were estimated to be about 49 kDa and 200 kDa, respectively, indicating that the enzyme was composed of four subunits of equal molecular masses. The Km and Vmax values of the enzyme for l-isoleucine were 5.00 mM and 153 ?mol·min?1·mg?1, respectively, and those for d-allo-isoleucine were 13.2 mM and 286 ?mol·min?1·mg?1, respectively. Hydroxylamine and other inhibitors of pyridoxal 5?-phosphate-dependent enzymes completely blocked the enzyme activity, indicating the enzyme requires pyridoxal 5?-phosphate as a coenzyme. This is the first evidence of an amino acid racemase that specifically catalyzes racemization of nonpolar amino acids at the C-2 position. PMID:24039265

  17. Probiotic and cultural characteristic of strain Lactobacillus gasseri 4/13 of human origin

    PubMed Central

    Baltova, Kalinka; Dimitrov, Zhechko

    2014-01-01

    Lactobacillus gasseri within the Lactobacillus acidophilus group is a major species in the human microflora. The potential probiotic properties of a L. gasseri strain of human origin were evaluated. Out of 17 studied L. gasseri strains, L. gasseri 4/13 showed the highest immunomodulatory effect (induction of interferon gamma measured by sandwich enzyme-linked immunosorbent assay) in Balb/c mouse splenocytes in vitro and the highest rate of adhesion to Caco-2 human epithelial cells. The strain also reduced the concentration of cholesterol in the growth medium by 65% as compared with the initial concentration (measured spectrophotometrically). These probiotic properties indicate that L. gasseri 4/13 could prove an attractive concentrated adjunct monoculture in the production of new functional foods. To obtain a freeze-dried bacterial concentrate from L. gasseri 4/13, the influence of different culture media, temperatures and pH values on the accumulation of cell biomass was studied. Yoghurt samples were produced using a classical fermentation technology. Freeze-dried concentrated monoculture of L. gasseri 4/13 with over 1 × 1010 CFU/g viable cells was added as an adjunct culture together with a starter. The viable L. gasseri 4/13 cells remained above the critical value of 106 CFU/g during storage at 5 °C for the entire 20-day period. Organoleptic tests did not reveal any adverse change in the product taste and aroma of yoghurt samples at the 20th day. In conclusion, L. gasseri 4/13 was selected as having suitable probiotic and cultural characteristics for production of fermented milk products with high nutritional and biological value.

  18. Highly Heterogeneous Probiotic Lactobacillus Species in Healthy Iranians with Low Functional Activities

    PubMed Central

    Rohani, Mahdi; Noohi, Nasrin; Talebi, Malihe; Katouli, Mohammad; Pourshafie, Mohammad R.

    2015-01-01

    Background Lactic acid bacteria (LAB) have been considered as potentially probiotic organisms due to their potential human health properties. This study aimed to evaluate both in vitro and in vivo, the potential probiotic properties of Lactobacillus species isolated from fecal samples of healthy humans in Iran. Methods and Results A total of 470 LAB were initially isolated from 53 healthy individual and characterized to species level. Of these, 88 (86%) were Lactobacillus species. Biochemical and genetic fingerprinting with Phene-Plate system (PhP-LB) and RAPD-PCR showed that the isolates were highly diverse consisted of 67(76.1%) and 75 (85.2%) single types (STs) and a diversity indices of 0.994 and 0.997, respectively. These strains were tested for production of adhesion to Caco-2 cells, antibacterial activity, production of B12, anti-proliferative effect and interleukin-8 induction on gut epithelial cell lines and antibiotic resistance against 9 commonly used antibiotics. Strains showing the characteristics consistent with probiotic strains, were further tested for their anti-inflammatory effect in mouse colitis model. Only one L. brevis; one L. rhamnosus and two L. plantarum were shown to have significant probiotic properties. These strains showed shortening the length of colon compared to dextran sulfate sodium and disease activity index (DAI) was also significantly reduced in mouse. Conclusion Low number of LAB with potential probiotic activity as well as high diversity of lactobacilli species was evident in Iranian population. It also suggest that specific strains of L. plantarum, L. brevis and L. rhamnosus with anti-inflammatory effect in mouse model of colitis could be used as a potential probiotic candidate in inflammatory bowel disease to decrease the disease activity index. PMID:26645292

  19. Three-component lysine/ornithine decarboxylation system in Lactobacillus saerimneri 30a.

    PubMed

    Romano, Andrea; Trip, Hein; Lolkema, Juke S; Lucas, Patrick M

    2013-03-01

    Lactic acid bacteria play a pivotal role in many food fermentations and sometimes represent a health threat due to the ability of some strains to produce biogenic amines that accumulate in foods and cause trouble following ingestion. These strains carry specific enzymatic systems catalyzing the uptake of amino acid precursors (e.g., ornithine and lysine), the decarboxylation inside the cell, and the release of the resulting biogenic amines (e.g., putrescine and cadaverine). This study aimed to identify the system involved in production of cadaverine from lysine, which has not been described to date for lactic acid bacteria. Strain Lactobacillus saerimneri 30a (formerly called Lactobacillus sp. 30a) produces both putrescine and cadaverine. The sequencing of its genome showed that the previously described ornithine decarboxylase gene was not associated with the gene encoding an ornithine/putrescine exchanger as in other bacteria. A new hypothetical decarboxylation system was detected in the proximity of the ornithine decarboxylase gene. It consisted of two genes encoding a putative decarboxylase sharing sequence similarities with ornithine decarboxylases and a putative amino acid transporter resembling the ornithine/putrescine exchangers. The two decarboxylases were produced in Escherichia coli, purified, and characterized in vitro, whereas the transporter was heterologously expressed in Lactococcus lactis and functionally characterized in vivo. The overall data led to the conclusion that the two decarboxylases and the transporter form a three-component decarboxylation system, with the new decarboxylase being a specific lysine decarboxylase and the transporter catalyzing both lysine/cadaverine and ornithine/putrescine exchange. To our knowledge, this is an unprecedented observation of a bacterial three-component decarboxylation system. PMID:23316036

  20. Bioactivity of proteins isolated from Lactobacillus plantarum L67 treated with Zanthoxylum piperitum DC glycoprotein.

    PubMed

    Song, S; Oh, S; Lim, K-T

    2015-06-01

    Lactobacilli in the human gastrointestinal tract have beneficial effects on the health of their host. To enhance these effects, the bioactivity of lactobacilli can be fortified through exogenous dietary or pharmacological agents, such as glycoproteins. To elucidate the inductive effect of Zanthoxylum piperitum DC (ZPDC) glycoprotein on Lactobacillus plantarum L67, we evaluated the radical-scavenging activity, anti-oxidative enzymes (SOD, GPx and CAT), growth rate, ATPase activity and ?-galactosidase activity of this strain. When Lact. plantarum L67 was treated with ZPDC glycoprotein at different concentrations, the intensities of a few SDS-PAGE bands were slightly changed. The amount of a 23 kDa protein was increased upon treatment with increasing concentrations of ZPDC glycoprotein. The results of this study indicate that the radical-scavenging activity for O2(-) and OH¯, but not for the DPPH radical, increased in a concentration-dependent manner after treatment with ZPDC glycoprotein. The activation of anti-oxidative enzymes (SOD, GPx and CAT), growth rate and ?-galactosidase activity also increased in a concentration-dependent manner in response to ZPDC glycoprotein treatment, whereas ATPase activity was decreased. In summary, ZPDC glycoprotein stimulated an increase in the bioactivity of Lact. plantarum L67. Significance and impact of the study: This study demonstrated that Lactobacillus plantarum L67 possesses anti-oxidative activity. This strain of lactic bacteria has been known to have various probiotic uses, such as yogurt starters and dietary additional supplements. We found, through this experiment, that the protein has a strong anti-oxidative character, and the activity can be enhanced by treatment with Zanthoxylum piperitum DC (ZPDC) glycoprotein. This study may be application of Lact. plantarum L67 treated by ZPDC glycoprotein in yogurt fermentation. It could be one of the avenues of minimizing yogurt postacidification during storage. In addition, it can be manufactured and incorporated in food products without losing viability and functionality of Lact. plantarum L67. PMID:25773674

  1. Therapeutic effects of Lactobacillus casei Qian treatment in activated carbon-induced constipated mice.

    PubMed

    Zhao, Xin; Suo, Hua-Yi; Qian, Yu; Li, Gui-Jie; Liu, Zhen-Hu; Li, Jian

    2015-08-01

    In the present study, the therapeutic effects of Lactobacillus casei Qian (LC-Qian), the key microorganism in Tibetan yak yoghurt, on activated carbon-induced constipation were determined in vivo. ICR mice were treated with LC-Qian for nine days by oral administration. The body weight, defecation status, gastrointestinal transit and defecation time of mice were assessed, and the serum levels of motilin (MTL), gastrin (Gas), endothelin (ET), somatostatin (SS), acetylcholinesterase (AChE), substance P (SP) and vasoactive intestinal peptide (VIP) were further evaluated. Bisacodyl was used as the positive control. The time until the first black stool defecation following carbon intake of the normal, control, 100 mg/kg bisacodyl-treated, Lactobacillus bulgaricus (LB)-treated, LC-Qian (L)-and LC-Qian (H)-treated mice was 93, 231, 121, 194, 172 and 157 min, respectively. Following treatment with LC-Qian, the gastrointestinal transit was increased to 52.4% [LC-Qian (L)] and 65.8% [LC-Qian (H)], while that in the group treated with the common lactic acid bacteria of LB was 40.3%. The MTL, Gas, ET, AChE, SP and VIP serum levels were significantly increased and levels of SS were reduced in mice following LC-Qian treatment compared with those in the control mice (P<0.05). Reverse transcription quantitative polymerase chain reaction indicated that LC-Qian raised the c-Kit, GDNF as well as SCF mRNA expression levels and reduced the TRPV1 and NOS expression levels in tissue of the small intestine in mice. These results suggested that lactic acid bacteria prevent constipation in mice, among which LC-Qian was the most effective. PMID:25955533

  2. Effect of Lactobacillus acidophilus NS1 on plasma cholesterol levels in diet-induced obese mice.

    PubMed

    Song, M; Park, S; Lee, H; Min, B; Jung, S; Park, S; Kim, E; Oh, S

    2015-03-01

    We investigated the probiotic properties of Lactobacillus acidophilus NS1, such as acid resistance, bile tolerance, adherence to HT-29 cells, and cholesterol assimilation activity. In an animal study, 7-wk-old male C57BL/6 mice were fed a normal diet, a high-fat diet (HFD), or an HFD with L. acidophilus NS1 (ca. 1.0×10(8) cfu/mL) for 10 wk. Total cholesterol and low-density lipoprotein (LDL) cholesterol levels were significantly lower in mice fed an HFD with L. acidophilus NS1 than in those fed an HFD only, whereas high-density lipoprotein cholesterol levels were similar between these 2 groups. To understand the mechanism of the cholesterol-lowering effect of L. acidophilus NS1 on the HFD-mediated increase in plasma cholesterol levels, we determined mRNA levels of genes involved in cholesterol homeostasis in the liver. Expression of sterol regulatory element-binding protein 2 (Srebp2) and LDL receptor (Ldlr) in the liver was dramatically reduced in mice fed a HFD compared with those fed a normal diet. When L. acidophilus NS1 was administered orally to HFD-fed mice, an HFD-induced suppression of Srebp2 and Ldlr expression in the liver was abolished. These results suggest that the oral administration of L. acidophilus NS1 to mice fed an HFD increased the expression of Srebp2 and Ldlr in the liver, which was inhibited by high fat intake, thus leading to a decrease in plasma cholesterol levels. Lactobacillus acidophilus NS1 could be a useful probiotic microorganism for cholesterol-lowering dairy products and the improvement of hyperlipidemia and hepatic lipid metabolism. PMID:25468703

  3. Solution Structure of Acidocin B, a Circular Bacteriocin Produced by Lactobacillus acidophilus M46

    PubMed Central

    Acedo, Jeella Z.; van Belkum, Marco J.; Lohans, Christopher T.; McKay, Ryan T.; Miskolzie, Mark

    2015-01-01

    Acidocin B, a bacteriocin produced by Lactobacillus acidophilus M46, was originally reported to be a linear peptide composed of 59 amino acid residues. However, its high sequence similarity to gassericin A, a circular bacteriocin from Lactobacillus gasseri LA39, suggested that acidocin B might be circular as well. Acidocin B was purified from culture supernatant by a series of hydrophobic interaction chromatographic steps. Its circular nature was ascertained by matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry and tandem mass spectrometry (MS/MS) sequencing. The peptide sequence was found to consist of 58 amino acids with a molecular mass of 5,621.5 Da. The sequence of the acidocin B biosynthetic gene cluster was also determined and showed high nucleotide sequence similarity to that of gassericin A. The nuclear magnetic resonance (NMR) solution structure of acidocin B in sodium dodecyl sulfate micelles was elucidated, revealing that it is composed of four ?-helices of similar length that are folded to form a compact, globular bundle with a central pore. This is a three-dimensional structure for a member of subgroup II circular bacteriocins, which are classified based on their isoelectric points of ?7 or lower. Comparison of acidocin B with carnocyclin A, a subgroup I circular bacteriocin with four ?-helices and a pI of 10, revealed differences in the overall folding. The observed variations could be attributed to inherent diversity in their physical properties, which also required the use of different solvent systems for three-dimensional structural elucidation. PMID:25681186

  4. Lactobacillus helveticus SBT2171, a cheese starter, regulates proliferation and cytokine production of immune cells.

    PubMed

    Yamashita, M; Ukibe, K; Uenishi, H; Hosoya, T; Sakai, F; Kadooka, Y

    2014-01-01

    Consumption of a Lactobacillus helveticus SBT2171 (LH2171)-containing cheese has been reported to exhibit immunoregulatory actions, including an increase in regulatory T cell population and reduction in proinflammatory cytokine production in mice. We examined the in vitro effects of LH2171 cells per se on immune cell function, specifically proliferation and cytokine production, which are primary reactions of the immune response. Immune cell fractions were prepared by mechanical disruption of mesenteric lymph nodes (MLN), Peyer's patches (PP), and spleens (SP) of mice. The cell fractions were dispensed into a culture plate and stimulated with anti-CD3/CD28 antibody beads in place of antigen-presenting cells or lipopolysaccharide (LPS) in the presence or absence of heat-treated LH2171 cells and other bacterial strains for comparison. After incubation, proliferation, cytokine production, and cell viability of the immune cells were determined. The LH2171 significantly inhibited the proliferation of MLN immune cells stimulated with anti-CD3/CD28 compared with other bacterial strains. The antiproliferative potency of LH2171 was effective not only on MLN but also on PP and SP stimulated with anti-CD3/CD28 or LPS. The LH2171 also decreased LPS-stimulated IL-6 production from MLN, PP, and SP, and IL-1? production from SP, but LH2171 did not affect the viability of immune cells. The LH2171 inhibited immune cell proliferation and proinflammatory cytokine (IL-6 and IL-1?) production. The inhibitory actions were not due to cytotoxicity to immune cells, suggesting that LH2171 is a dairy Lactobacillus strain with beneficial immunoregulatory properties. PMID:24881786

  5. Indication for Co-evolution of Lactobacillus johnsonii with its hosts

    PubMed Central

    2012-01-01

    Background The intestinal microbiota, composed of complex bacterial populations, is host-specific and affected by environmental factors as well as host genetics. One important bacterial group is the lactic acid bacteria (LAB), which include many health-promoting strains. Here, we studied the genetic variation within a potentially probiotic LAB species, Lactobacillus johnsonii, isolated from various hosts. Results A wide survey of 104 fecal samples was carried out for the isolation of L. johnsonii. As part of the isolation procedure, terminal restriction fragment length polymorphism (tRFLP) was performed to identify L. johnsonii within a selected narrow spectrum of fecal LAB. The tRFLP results showed host specificity of two bacterial species, the Enterococcus faecium species cluster and Lactobacillus intestinalis, to different host taxonomic groups while the appearance of L. johnsonii and E. faecalis was not correlated with any taxonomic group. The survey ultimately resulted in the isolation of L. johnsonii from few host species. The genetic variation among the 47?L. johnsonii strains isolated from the various hosts was analyzed based on variation at simple sequence repeats (SSR) loci and multi-locus sequence typing (MLST) of conserved hypothetical genes. The genetic relationships among the strains inferred by each of the methods were similar, revealing three different clusters of L. johnsonii strains, each cluster consisting of strains from a different host, i.e. chickens, humans or mice. Conclusions Our typing results support phylogenetic separation of L. johnsonii strains isolated from different animal hosts, suggesting specificity of L. johnsonii strains to their hosts. Taken together with the tRFLP results, that indicated the association of specific LAB species with the host taxonomy, our study supports co-evolution of the host and its intestinal lactic acid bacteria. PMID:22827843

  6. Arabinoxylan oligosaccharide hydrolysis by family 43 and 51 glycosidases from Lactobacillus brevis DSM 20054.

    PubMed

    Michlmayr, Herbert; Hell, Johannes; Lorenz, Cindy; Böhmdorfer, Stefan; Rosenau, Thomas; Kneifel, Wolfgang

    2013-11-01

    Due to their potential prebiotic properties, arabinoxylan-derived oligosaccharides [(A)XOS] are of great interest as functional food and feed ingredients. While the (A)XOS metabolism of Bifidobacteriaceae has been extensively studied, information regarding lactic acid bacteria (LAB) is still limited in this context. The aim of the present study was to fill this important gap by characterizing candidate (A)XOS hydrolyzing glycoside hydrolases (GHs) identified in the genome of Lactobacillus brevis DSM 20054. Two putative GH family 43 xylosidases (XynB1 and XynB2) and a GH family 43 arabinofuranosidase (Abf3) were heterologously expressed and characterized. While the function of XynB1 remains unclear, XynB2 could efficiently hydrolyze xylooligosaccharides. Abf3 displayed high specific activity for arabinobiose but could not release arabinose from an (A)XOS preparation. However, two previously reported GH 51 arabinofuranosidases from Lb. brevis were able to specifically remove ?-1,3-linked arabinofuranosyl residues from arabino-xylooligosaccharides (AXHm3 specificity). These results imply that Lb. brevis is at least genetically equipped with functional enzymes in order to hydrolyze the depolymerization products of (arabino)xylans and arabinans. The distribution of related genes in Lactobacillales genomes indicates that GH 43 and, especially, GH 51 glycosidase genes are rare among LAB and mainly occur in obligately heterofermentative Lactobacillus spp., Pediococcus spp., members of the Leuconostoc/Weissella branch, and Enterococcus spp. Apart from the prebiotic viewpoint, this information also adds new perspectives on the carbohydrate (i.e., pentose-oligomer) metabolism of LAB species involved in the fermentation of hemicellulose-containing substrates. PMID:23995921

  7. Therapeutic effect of activated carbon-induced constipation mice with Lactobacillus fermentum Suo on treatment.

    PubMed

    Suo, Huayi; Zhao, Xin; Qian, Yu; Li, Guijie; Liu, Zhenhu; Xie, Jie; Li, Jian

    2014-01-01

    The aim of this study was to investigate the effects of Lactobacillus fermentum Suo (LF-Suo) on activated carbon-induced constipation in ICR (Institute of Cancer Research) mice. ICR mice were orally administered with lactic acid bacteria for 9 days. Body weight, diet intake, drinking amount, defecation status, gastrointestinal transit and defecation time, and the serum levels of MTL (motilin), Gas (gastrin), ET (endothelin), SS (somatostatin), AChE (acetylcholinesterase), SP (substance P), VIP (vasoactive intestinal peptide) were used to evaluate the preventive effects of LF-Suo on constipation. Bisacodyl, a laxative drug, was used as a positive control. The normal, control, 100 mg/kg bisacodyl treatment, LB (Lactobacillus bulgaricus)-, LF-Suo (L)- and LF-Suo (H)-treated mice showed the time to the first black stool defecation at 90, 218, 117, 180, 155 and 137 min, respectively. By the oral administration of LB-, LF-Suo (L), LF-Suo (H) or bisacodyl (100 mg/kg), the gastrointestinal transit was reduced to 55.2%, 72.3%, 85.5% and 94.6%, respectively, of the transit in normal mice, respectively. In contrast to the control mice, the serum levels of MTL, Gas, ET, AChE, SP and VIP were significantly increased and the serum levels of SS were reduced in the mice treated with LF-Suo (p < 0.05). By the RT-PCR (reverse transcription-polymerase chain reaction) and western blot assays, LF-Suo increased the c-Kit, SCF (stem cell factor), GDNF (glial cell line-derived neurotrophic factor) and decreased TRPV1 (transient receptor potential vanilloid 1), NOS (nitric oxide synthase) expressions of small intestine tissue in mice. These results demonstrate that lactic acid bacteria has preventive effects on mouse constipation and LF-Suo demonstrated the best functional activity. PMID:25464378

  8. Preventive effect of Lactobacillus fermentum Lee on activated carbon-induced constipation in mice.

    PubMed

    Qian, Yu; Suo, Huayi; DU, Muying; Zhao, Xin; Li, Jian; Li, Gui-Jie; Song, Jia-Li; Liu, Zhenhu

    2015-01-01

    The aim of this study was to investigate the effects of Lactobacillus fermentum Lee (LF-Lee) on activated carbon-induced constipation in ICR mice. ICR mice were orally administered lactic acid bacteria for nine days. Body weight, dietary and water intake, defecation status, gastrointestinal (GI) transit and defecation time, as well as levels of motilin (MTL), gastrin (Gas), endothelin (ET), somatostatin (SS), acetylcholinesterase (AChE), substance P (SP) and vasoactive intestinal peptide (VIP) in serum were measured to evaluate the preventive effects of LF-Lee on constipation. Bisacodyl, a laxative drug, was administered as a positive control. The time taken until the first defecation of a black stool for normal, control, bisacodyl- (100 mg/kg, oral administration), Lactobacillus bulgaricus (LB)-, LF-Lee low dose (L)- and LF-Lee high dose (H)-treated mice was 90, 218, 117, 180, 161 and 151 min, respectively. Following the consumption of LB, LF-Lee (L) or LF-Lee (H), or the oral administration of bisacodyl, the GI transit was reduced to 55.2, 65.8, 73.1 and 94.6%, respectively, of the transit in normal mice. The serum levels of MTL, Gas, ET, AChE, SP and VIP were significantly increased and those of SS were reduced in the mice treated with LF-Lee compared with those in the untreated control mice (P<0.05). These results demonstrate that lactic acid bacteria have preventive effects on constipation in mice and that LF-Lee has superior functional activity. PMID:25452815

  9. Lactobacillus crustorum KH: novel prospective probiotic strain isolated from Iranian traditional dairy products.

    PubMed

    Sharafi, Hakimeh; Derakhshan, Venos; Paknejad, Mojgan; Alidoust, Leila; Tohidi, Azadeh; Pornour, Majid; Hajfarajollah, Hamidreza; Zahiri, Hossein Shahbani; Noghabi, Kambiz Akbari

    2015-02-01

    In recent years, exploring novel probiotic strains for therapeutic intervention has been raised due to the significant increase in market demand. This study aimed to investigate the certain probiotic properties of 15 Lactobacillus isolates from Iranian traditional dairy products. Among them, a novel potential probiotic strain was isolated and identified as Lactobacillus crustorum. The characteristics of potential probiotics were examined in terms of resistance to acidity, bile, and salinity as well as antibiotic tolerance and antibacterial activity. L. crustorum KH has shown tolerance property to bile (0.3 % w), acidity (pH 2-9), and salinity (1-5 % NaCl) and strong antibacterial activity against tested enteropathogens by well-diffusion assay. Furthermore, in vivo study and histological assays were performed to study whether live and heat-killed cells of L. crustorum KH are able to protect against the challenge of Escherichia coli O157:H7 in the gastrointestinal tract of mice used as an experimental model. Therefore, heat-killed and live cells of L. crustorum KH were inoculated by gavage to different groups of 4-6-week-old female BALB/c mice in doses of 10(8) colony-forming unit (CFU)/dose. Thereafter, these mice were challenged with E. coli O157:H7 also inoculated in the gastrointestinal tract (GIT) of the animals. The results showed that heat-killed cells of L. crustorum KH exert a protective effect against E. coli O157:H7 colonization at different degrees, being lower than that produced by viable cells. PMID:25467945

  10. Identification of a Putative Operon Involved in Fructooligosaccharide Utilization by Lactobacillus paracasei? †

    PubMed Central

    Goh, Yong Jun; Zhang, Chaomei; Benson, Andrew K.; Schlegel, Vicki; Lee, Jong-Hwa; Hutkins, Robert W.

    2006-01-01

    The growth and activity of some Lactobacillus and Bifidobacterium strains are stimulated by the presence of nondigestible fructooligosaccharides (FOS), which are selectively fermented by specific intestinal bacteria. Consumption of FOS, therefore, enriches for those bacteria that possess metabolic pathways necessary for FOS metabolism. In this study, a DNA microarray consisting of 7,680 random genomic library fragments of Lactobacillus paracasei 1195 was used to examine genes involved in the utilization of FOS in this organism. Differential expression profiles between cells grown on FOS and those grown on glucose provided a basis for identifying genes specifically induced by FOS. Several of the FOS-induced genes shared sequence identity with genes encoding ?-fructosidases and components of phosphoenolpyruvate-dependent phosphotransferase systems (PTS). These genes were organized in a putative operon, designated the fos operon, that may play an essential role in FOS utilization. The complete 7,631-bp nucleotide sequence of the putative fos operon was determined and consists of fosABCDXE genes, which encode a putative fructose/mannose PTS (FosABCDX) and a ?-fructosidase precursor (FosE). The latter contains an N-terminal signal peptide sequence and cell wall sorting signals at the C-terminal region, suggesting its localization at the cell wall. Inactivation of the fosE gene led to impaired growth on FOS and other ?-fructose-linked carbohydrates. Transcriptional analysis by reverse transcriptase PCR suggested that fosABCDXE was cotranscribed as a single mRNA during growth on FOS. Expression array analysis revealed that when glucose was added to FOS-grown cells, transcription of the FOS-induced genes was repressed, indicating that FOS metabolism is subject to catabolite regulation. PMID:17028235

  11. Purification and Characterization of Novel Antifungal Compounds from the Sourdough Lactobacillus plantarum Strain 21B

    PubMed Central

    Lavermicocca, Paola; Valerio, Francesca; Evidente, Antonio; Lazzaroni, Silvia; Corsetti, Aldo; Gobbetti, Marco

    2000-01-01

    Sourdough lactic acid bacteria were selected for antifungal activity by a conidial germination assay. The 10-fold-concentrated culture filtrate of Lactobacillus plantarum 21B grown in wheat flour hydrolysate almost completely inhibited Eurotium repens IBT18000, Eurotium rubrum FTDC3228, Penicillium corylophilum IBT6978, Penicillium roqueforti IBT18687, Penicillium expansum IDM/FS2, Endomyces fibuliger IBT605 and IDM3812, Aspergillus niger FTDC3227 and IDM1, Aspergillus flavus FTDC3226, Monilia sitophila IDM/FS5, and Fusarium graminearum IDM623. The nonconcentrated culture filtrate of L. plantarum 21B grown in whole wheat flour hydrolysate had similar inhibitory activity. The activity was fungicidal. Calcium propionate at 3 mg ml?1 was not effective under the same assay conditions, while sodium benzoate caused inhibition similar to L. plantarum 21B. After extraction with ethyl acetate, preparative silica gel thin-layer chromatography, and chromatographic and spectroscopic analyses, novel antifungal compounds such as phenyllactic and 4-hydroxy-phenyllactic acids were identified in the culture filtrate of L. plantarum 21B. Phenyllactic acid was contained at the highest concentration in the bacterial culture filtrate and had the highest activity. It inhibited all the fungi tested at a concentration of 50 mg ml?1 except for P. roqueforti IBT18687 and P. corylophilum IBT6978 (inhibitory concentration, 166 mg ml?1). L. plantarum 20B, which showed high antimold activity, was also selected. Preliminary studies showed that phenyllactic and 4-hydroxy-phenyllactic acids were also contained in the bacterial culture filtrate of strain 20B. Growth of A. niger FTDC3227 occurred after 2 days in breads started with Saccharomyces cerevisiae 141 alone or with S. cerevisiae and Lactobacillus brevis 1D, an unselected but acidifying lactic acid bacterium, while the onset of fungal growth was delayed for 7 days in bread started with S. cerevisiae and selected L. plantarum 21B. PMID:10966432

  12. Spontaneously Induced Prophages in Lactobacillus gasseri Contribute to Horizontal Gene Transfer

    PubMed Central

    Baugher, J. L.; Durmaz, E.

    2014-01-01

    Lactobacillus gasseri is an endogenous species of the human gastrointestinal tract and vagina. With recent advances in microbial taxonomy, phylogenetics, and genomics, L. gasseri is recognized as an important commensal and is increasingly being used in probiotic formulations. L. gasseri strain ADH is lysogenic and harbors two inducible prophages. In this study, prophage ?adh was found to spontaneously induce in broth cultures to populations of ?107 PFU/ml by stationary phase. The ?adh prophage-cured ADH derivative NCK102 was found to harbor a new, second inducible phage, vB_Lga_jlb1 (jlb1). Phage jlb1 was sequenced and found to be highly similar to the closely related phage LgaI, which resides as two tandem prophages in the neotype strain L. gasseri ATCC 33323. The common occurrence of multiple prophages in L. gasseri genomes, their propensity for spontaneous induction, and the high degree of homology among phages within multiple species of Lactobacillus suggest that temperate bacteriophages likely contribute to horizontal gene transfer (HGT) in commensal lactobacilli. In this study, the host ranges of phages ?adh and jlb1 were determined against 16 L. gasseri strains. The transduction range and the rate of spontaneous transduction were investigated in coculture experiments to ascertain the degree to which prophages can promote HGT among a variety of commensal and probiotic lactobacilli. Both ?adh and jlb1 particles were confirmed to mediate plasmid transfer. As many as ?103 spontaneous transductants/ml were obtained. HGT by transducing phages of commensal lactobacilli may have a significant impact on the evolution of bacteria within the human microbiota. PMID:24682298

  13. Bacteriophage-encoded lytic enzymes control growth of contaminating Lactobacillus found in fuel ethanol fermentations

    PubMed Central

    2013-01-01

    Background Reduced yields of ethanol due to bacterial contamination in fermentation cultures weaken the economics of biofuel production. Lactic acid bacteria are considered the most problematic, and surveys of commercial fuel ethanol facilities have found that species of Lactobacillus are predominant. Bacteriophage lytic enzymes are peptidoglycan hydrolases that can degrade the Gram positive cell wall when exposed externally and provide a novel source of antimicrobials that are highly refractory to resistance development. Results The streptococcal phage LambdaSa2 (?Sa2) endolysin demonstrated strong lytic activity towards 17 of 22 strains of lactobacilli, staphylococci or streptococci and maintained an optimal specific activity at pH 5.5 and in the presence of ? 5% ethanol (fermentation conditions) toward L. fermentum. Lactobacillus bacteriophage endolysins LysA, LysA2 and LysgaY showed exolytic activity towards 60% of the lactobacilli tested including four L. fermentum isolates from fuel ethanol fermentations. In turbidity reduction assays LysA was able to reduce optical density >75% for 50% of the sensitive strains and >50% for the remaining strains. LysA2 and LysgaY were only able to decrease cellular turbidity by <50%. Optimal specific activities were achieved for LysA, LysA2, and LysgaY at pH 5.5. The presence of ethanol (?5%) did not reduce the lytic activity. Lysins were able to reduce both L. fermentum (BR0315-1) (?Sa2 endolysin) and L. reuteri (B-14171) (LysA) contaminants in mock fermentations of corn fiber hydrolysates. Conclusion Bacteriophage lytic enzymes are strong candidates for application as antimicrobials to control lactic acid bacterial contamination in fuel ethanol fermentations. PMID:23390890

  14. Identification of Lactobacillus plantarum genes modulating the cytokine response of human peripheral blood mononuclear cells

    PubMed Central

    2010-01-01

    Background Modulation of the immune system is one of the most plausible mechanisms underlying the beneficial effects of probiotic bacteria on human health. Presently, the specific probiotic cell products responsible for immunomodulation are largely unknown. In this study, the genetic and phenotypic diversity of strains of the Lactobacillus plantarum species were investigated to identify genes of L. plantarum with the potential to influence the amounts of cytokines interleukin 10 (IL-10) and IL-12 and the ratio of IL-10/IL-12 produced by peripheral blood mononuclear cells (PBMCs). Results A total of 42 Lactobacillus plantarum strains isolated from diverse environmental and human sources were evaluated for their capacity to stimulate cytokine production in PBMCs. The L. plantarum strains induced the secretion of the anti-inflammatory cytokine IL-10 over an average 14-fold range and secretion of the pro-inflammatory cytokine IL-12 over an average 16-fold range. Comparisons of the strain-specific cytokine responses of PBMCs to comparative genome hybridization profiles obtained with L. plantarum WCFS1 DNA microarrays (also termed gene-trait matching) resulted in the identification of 6 candidate genetic loci with immunomodulatory capacities. These loci included genes encoding an N-acetyl-glucosamine/galactosamine phosphotransferase system, the LamBDCA quorum sensing system, and components of the plantaricin (bacteriocin) biosynthesis and transport pathway. Deletion of these genes in L. plantarum WCFS1 resulted in growth phase-dependent changes in the PBMC IL-10 and IL-12 cytokine profiles compared with wild-type cells. Conclusions The altered PBMC cytokine profiles obtained with the L. plantarum WCFS1 mutants were in good agreement with the predictions made by gene-trait matching for the 42 L. plantarum strains. This study therefore resulted in the identification of genes present in certain strains of L. plantarum which might be responsible for the stimulation of anti- or pro-inflammatory immune responses in the gut. PMID:21080958

  15. Preventive effect of Lactobacillus fermentum Lee on activated carbon-induced constipation in mice

    PubMed Central

    QIAN, YU; SUO, HUAYI; DU, MUYING; ZHAO, XIN; LI, JIAN; LI, GUI-JIE; SONG, JIA-LI; LIU, ZHENHU

    2015-01-01

    The aim of this study was to investigate the effects of Lactobacillus fermentum Lee (LF-Lee) on activated carbon-induced constipation in ICR mice. ICR mice were orally administered lactic acid bacteria for nine days. Body weight, dietary and water intake, defecation status, gastrointestinal (GI) transit and defecation time, as well as levels of motilin (MTL), gastrin (Gas), endothelin (ET), somatostatin (SS), acetylcholinesterase (AChE), substance P (SP) and vasoactive intestinal peptide (VIP) in serum were measured to evaluate the preventive effects of LF-Lee on constipation. Bisacodyl, a laxative drug, was administered as a positive control. The time taken until the first defecation of a black stool for normal, control, bisacodyl- (100 mg/kg, oral administration), Lactobacillus bulgaricus (LB)-, LF-Lee low dose (L)- and LF-Lee high dose (H)-treated mice was 90, 218, 117, 180, 161 and 151 min, respectively. Following the consumption of LB, LF-Lee (L) or LF-Lee (H), or the oral administration of bisacodyl, the GI transit was reduced to 55.2, 65.8, 73.1 and 94.6%, respectively, of the transit in normal mice. The serum levels of MTL, Gas, ET, AChE, SP and VIP were significantly increased and those of SS were reduced in the mice treated with LF-Lee compared with those in the untreated control mice (P<0.05). These results demonstrate that lactic acid bacteria have preventive effects on constipation in mice and that LF-Lee has superior functional activity. PMID:25452815

  16. Characterization of a Mobile clpL Gene from Lactobacillus rhamnosus

    PubMed Central

    Suokko, Aki; Savijoki, Kirsi; Malinen, Erja; Palva, Airi; Varmanen, Pekka

    2005-01-01

    Two genes encoding ClpL ATPase proteins were identified in a probiotic Lactobacillus rhamnosus strain, E-97800. Sequence analyses revealed that the genes, designated clpL1 and clpL2, share 80% identity. The clpL2 gene showed the highest degree of identity (98.5%) to a clpL gene from Lactobacillus plantarum WCFSI, while it was not detected in three other L. rhamnosus strains studied. According to Northern analyses, the expression of clpL1 and the clpL2 were induced during heat shock by >20- and 3-fold, respectively. The functional promoter regions were determined by primer extension analyses, and the clpL1 promoter was found to be overlapped by an inverted repeat structure identical to the conserved CIRCE element, indicating that clpL1 belongs to the HrcA regulon in L. rhamnosus. No consensus binding sites for HrcA or CtsR could be identified in the clpL2 promoter region. Interestingly, the clpL2 gene was found to be surrounded by truncated transposase genes and flanked by inverted repeat structures nearly identical to the terminal repeats of the ISLpl1 from L. plantarum HN38. Furthermore, clpL2 was shown to be mobilized during prolonged cultivation at elevated temperature. The presence of a gene almost identical to clpL2 in L. plantarum and its absence in other L. rhamnosus strains suggest that the L. rhamnosus E-97800 has acquired the clpL2 gene via horizontal transfer. No change in the stress tolerance of the ClpL2-deficient derivative of E-97800 compared to the parental strain was observed. PMID:15812039

  17. Characterization of the melA Locus for ?-Galactosidase in Lactobacillus plantarum

    PubMed Central

    Silvestroni, Aurelio; Connes, Cristelle; Sesma, Fernando; Savoy de Giori, Graciela; Piard, Jean-Christophe

    2002-01-01

    Alpha-galactosides are abundant sugars in legumes such as soy. Because of the lack of ?-galactosidase (?-Gal) in the digestive tract, humans are unable to digest these sugars, which consequently induce flatulence. To develop the consumption of the otherwise highly nutritional soy products, the use of exogenous ?-Gal is promising. In this framework, we characterized the melA gene for ?-Gal in Lactobacillus plantarum. The melA gene encodes a cytoplasmic 84-kDa protein whose enzymatically active form occurs as oligomers. The melA gene was cloned and expressed in Escherichia coli, yielding an active ?-Gal. We show that melA is transcribed from its own promoter, yielding a monocistronic mRNA, and that it is regulated at the transcriptional level, i.e., it is induced by melibiose but is not totally repressed by glucose. Posttranscriptional regulation by the carbon source could also occur. Upstream of melA, a putative galactoside transporter, designated RafP, was identified that shows high homology to LacS, the unique transporter for both ?- and ?-galactosides in Streptococcus thermophilus. rafP is also expressed as a monocistronic mRNA. Downstream of melA, the lacL and lacM genes were identified that encode a heterodimeric ?-galactosidase. A putative galM gene identified in the same cluster suggests the presence of a galactose operon. These results indicate that the genes involved in galactoside catabolism are clustered in L. plantarum ATCC 8014. This first genetic characterization of melA and of its putative associated transporter, rafP, in a lactobacillus opens doors to various applications both in the manufacture of soy-derived products and in probiotic and nutraceutical issues. PMID:12406739

  18. Characterization of the temperate bacteriophage phi adh and plasmid transduction in Lactobacillus acidophilus ADH.

    PubMed Central

    Raya, R R; Kleeman, E G; Luchansky, J B; Klaenhammer, T R

    1989-01-01

    Lactobacillus acidophilus ADH is lysogenic and harbors an inducible prophage, phi adh. Bacteriophage were detected in cell lysates induced by treatment with mitomycin C or UV light. Electron microscopy of lysates revealed phage particles with a hexagonal head (62 nm) and a long, noncontractile, flexible tail (398 nm) ending in at last five short fibers. Phage phi adh was classified within Bradley's B1 phage group and the Siphoviridae family. The phi adh genome is a linear double-stranded DNA molecule of 41.7 kilobase pairs with cohesive ends: a physical map of the phi adh genome was constructed. A prophage-cured derivative of strain ADH, designated NCK102, was isolated from cells that survived UV exposure. NCK102 did not exhibit mitomycin C-induced lysis, but broth cultures lysed upon addition of phage. Phage phi adh produced clear plaques on NCK102 in media containing 10 mM CaCl2 at pH values between 5.2 and 5.5. A relysogenized derivative (NCK103) of NCK102 was isolated that exhibited mitomycin C-induced lysis and superinfection immunity to phage phi adh. Hybridization experiments showed that the phi adh genome was present in the ADH and NCK103 chromosomes, but absent in NCK102. These results demonstrated classic lytic and lysogenic cycles of replication for the temperate phage phi adh induced from L. acidophilus ADH. Phage phi adh also mediates transduction of plasmid DNA. Transductants of strain ADH containing pC194, pGK12, pGB354, and pVA797 were detected at frequencies in the range of 3.6 x 10(-8) to 8.3 x 10(-10) per PFU. Rearrangements or deletions were not detected in these plasmids as a consequence of transduction. This is the first description of plasmid transduction in the genus Lactobacillus. Images PMID:2508554

  19. Therapeutic Effect of Activated Carbon-Induced Constipation Mice with Lactobacillus fermentum Suo on Treatment

    PubMed Central

    Suo, Huayi; Zhao, Xin; Qian, Yu; Li, Guijie; Liu, Zhenhu; Xie, Jie; Li, Jian

    2014-01-01

    The aim of this study was to investigate the effects of Lactobacillus fermentum Suo (LF-Suo) on activated carbon-induced constipation in ICR (Institute of Cancer Research) mice. ICR mice were orally administered with lactic acid bacteria for 9 days. Body weight, diet intake, drinking amount, defecation status, gastrointestinal transit and defecation time, and the serum levels of MTL (motilin), Gas (gastrin), ET (endothelin), SS (somatostatin), AChE (acetylcholinesterase), SP (substance P), VIP (vasoactive intestinal peptide) were used to evaluate the preventive effects of LF-Suo on constipation. Bisacodyl, a laxative drug, was used as a positive control. The normal, control, 100 mg/kg bisacodyl treatment, LB (Lactobacillus bulgaricus)-, LF-Suo (L)- and LF-Suo (H)-treated mice showed the time to the first black stool defecation at 90, 218, 117, 180, 155 and 137 min, respectively. By the oral administration of LB-, LF-Suo (L), LF-Suo (H) or bisacodyl (100 mg/kg), the gastrointestinal transit was reduced to 55.2%, 72.3%, 85.5% and 94.6%, respectively, of the transit in normal mice, respectively. In contrast to the control mice, the serum levels of MTL, Gas, ET, AChE, SP and VIP were significantly increased and the serum levels of SS were reduced in the mice treated with LF-Suo (p < 0.05). By the RT-PCR (reverse transcription–polymerase chain reaction) and western blot assays, LF-Suo increased the c-Kit, SCF (stem cell factor), GDNF (glial cell line-derived neurotrophic factor) and decreased TRPV1 (transient receptor potential vanilloid 1), NOS (nitric oxide synthase) expressions of small intestine tissue in mice. These results demonstrate that lactic acid bacteria has preventive effects on mouse constipation and LF-Suo demonstrated the best functional activity. PMID:25464378

  20. Mucosal Immunogenicity of Genetically Modified Lactobacillus acidophilus Expressing an HIV-1 Epitope within the Surface Layer Protein

    PubMed Central

    Kajikawa, Akinobu; Zhang, Lin; LaVoy, Alora; Bumgardner, Sara; Klaenhammer, Todd R.; Dean, Gregg A.

    2015-01-01

    Surface layer proteins of probiotic lactobacilli are theoretically efficient epitope-displaying scaffolds for oral vaccine delivery due to their high expression levels and surface localization. In this study, we constructed genetically modified Lactobacillus acidophilus strains expressing the membrane proximal external region (MPER) from human immunodeficiency virus type 1 (HIV-1) within the context of the major S-layer protein, SlpA. Intragastric immunization of mice with the recombinants induced MPER-specific and S-layer protein-specific antibodies in serum and mucosal secretions. Moreover, analysis of systemic SlpA-specific cytokines revealed that the responses appeared to be Th1 and Th17 dominant. These findings demonstrated the potential use of the Lactobacillus S-layer protein for development of oral vaccines targeting specific peptides. PMID:26509697

  1. Effects of fermentation with Lactobacillus rhamnosus GG on product quality and fatty acids of goat milk yogurt.

    PubMed

    Jia, Ru; Chen, Han; Chen, Hui; Ding, Wu

    2016-01-01

    The effect of fermentation with Lactobacillus rhamnosus GG on the product quality of goat milk yogurt using traditional yogurt starter was studied through single-factor experiments and orthogonal experiments. The optimum fermentation condition was evaluated by the titratable acidity of goat milk yogurt, water-retaining capability, sensory score, and texture properties; the fatty acids of the fermented goat milk were determined by a gas chromatograph. Results indicate that high product quality of goat milk yogurt can be obtained and the content of short-chain and medium-chain fatty acids can be decreased significantly when amount of sugar added was 7%, inoculation amount was 3%, the ratio of 3 lactic acid bacteria-Lactobacillus delbrueckii ssp. bulgaricus, Streptococcus thermophilus, and L. rhamnosus GG-was 1:1:3, and fermentation temperature was 42°C. PMID:26601583

  2. Complete genome sequences and comparative genome analysis of Lactobacillus plantarum strain 5-2 isolated from fermented soybean.

    PubMed

    Liu, Chen-Jian; Wang, Rui; Gong, Fu-Ming; Liu, Xiao-Feng; Zheng, Hua-Jun; Luo, Yi-Yong; Li, Xiao-Ran

    2015-12-01

    Lactobacillus plantarum is an important probiotic and is mostly isolated from fermented foods. We sequenced the genome of L. plantarum strain 5-2, which was derived from fermented soybean isolated from Yunnan province, China. The strain was determined to contain 3114 genes. Fourteen complete insertion sequence (IS) elements were found in 5-2 chromosome. There were 24 DNA replication proteins and 76 DNA repair proteins in the 5-2 genome. Consistent with the classification of L. plantarum as a facultative heterofermentative lactobacillus, the 5-2 genome encodes key enzymes required for the EMP (Embden-Meyerhof-Parnas) and phosphoketolase (PK) pathways. Several components of the secretion machinery are found in the 5-2 genome, which was compared with L. plantarum ST-III, JDM1 and WCFS1. Most of the specific proteins in the four genomes appeared to be related to their prophage elements. PMID:26212213

  3. PRODUCTION OF MANNITOL FROM INULIN BY SIMULTANEOUS ENZYMATIC SACCHARIFICATION AND FERMENTATION WITH LACTOBACILLUS INTERMEDIUS NRRL B-3693

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The production of mannitol by Lactobacillus intermedius NRRL B-3693, using inulin as a substrate, was evaluated at pH 5.0 and 37 deg C. The bacterium produced mannitol (106.2 ± 0.3 g/l) from dilute acid hydrolyzate (pH 2.0, 121 deg C, 15 min) of inulin (150 g/L) in 34 h. It also produced mannitol ...

  4. Altered transcription of murine genes induced in the small bowel by administration of probiotic strain Lactobacillus rhamnosus HN001.

    PubMed

    Tannock, Gerald W; Taylor, Corinda; Lawley, Blair; Loach, Diane; Gould, Maree; Dunn, Amy C; McLellan, Alexander D; Black, Michael A; McNoe, Les; Dekker, James; Gopal, Pramod; Collett, Michael A

    2014-05-01

    Lactobacillus rhamnosus HN001 is a probiotic strain reported to increase resistance to epithelium-adherent and -invasive intestinal pathogens in experimental animals. To increase understanding of the relationship between strain HN001 and the bowel, transcription of selected genes in the mucosa of the murine small bowel was measured. Mice previously naive to lactobacilli (Lactobacillus-free mice) were examined after daily exposure to HN001 in drinking water. Comparisons were made to results from matched Lactobacillus-free mice. Infant and adult mice were investigated to provide a temporal view of gene expression in response to exposure to HN001. Genes sgk1, angptl4, and hspa1b, associated with the apoptosis pathway, were selected for investigation by reverse transcription-quantitative PCR on the basis of a preliminary duodenal DNA microarray screen. Normalized to gapdh gene transcription, these three genes were upregulated after 6 to 10 days exposure of adult mice to HN001. Angptl4 was shown by immunofluorescence to be upregulated in duodenal epithelial cells of mucosal samples. Epithelial cell migration was faster in HN001-exposed mice than in the Lactobacillus-free controls. Transcriptional responses in infant mice differed according to bowel region and age. For example, sgk1 was upregulated in duodenal, jejunal, and ileal mucosa of mice less than 25 days old, whereas angptl4 and hspa1b were upregulated at 10 days in the duodenum but downregulated in the jejunal mucosa until mice were 25 days old. Overall, the results provide links between a probiotic strain, mucosal gene expression, and host phenotype, which may be useful in delineating mechanisms of probiotic action. PMID:24584241

  5. Multilocus Sequence Typing of Lactobacillus casei Reveals a Clonal Population Structure with Low Levels of Homologous Recombination? †

    PubMed Central

    Diancourt, Laure; Passet, Virginie; Chervaux, Christian; Garault, Peggy; Smokvina, Tamara; Brisse, Sylvain

    2007-01-01

    Robust genotyping methods for Lactobacillus casei are needed for strain tracking and collection management, as well as for population biology research. A collection of 52 strains initially labeled L. casei or Lactobacillus paracasei was first subjected to rplB gene sequencing together with reference strains of Lactobacillus zeae, Lactobacillus rhamnosus, and other species. Phylogenetic analysis showed that all 52 strains belonged to a single compact L. casei-L. paracasei sequence cluster, together with strain CIP107868 (= ATCC 334) but clearly distinct from L. rhamnosus and from a cluster with L. zeae and CIP103137T (= ATCC 393T). The strains were genotyped using amplified fragment length polymorphism, multilocus sequence typing based on internal portions of the seven housekeeping genes fusA, ileS, lepA, leuS, pyrG, recA, and recG, and tandem repeat variation (multilocus variable-number tandem repeats analysis [MLVA] using nine loci). Very high concordance was found between the three methods. Although amounts of nucleotide variation were low for the seven genes (? ranging from 0.0038 to 0.0109), 3 to 12 alleles were distinguished, resulting in 31 sequence types. One sequence type (ST1) was frequent (17 strains), but most others were represented by a single strain. Attempts to subtype ST1 strains by MLVA, ribotyping, clustered regularly interspaced short palindromic repeat characterization, and single nucleotide repeat variation were unsuccessful. We found clear evidence for homologous recombination during the diversification of L. casei clones, including a putative intragenic import of DNA into one strain. Nucleotides were estimated to change four times more frequently by recombination than by mutation. However, statistical congruence between individual gene trees was retained, indicating that recombination is not frequent enough to disrupt the phylogenetic signal. The developed multilocus sequence typing scheme should be useful for future studies of L. casei strain diversity and evolution. PMID:17704267

  6. Dietary Enterococcus faecalis LAB31 Improves Growth Performance, Reduces Diarrhea, and Increases Fecal Lactobacillus Number of Weaned Piglets

    PubMed Central

    Hu, Yuanliang; Dun, Yaohao; Li, Shenao; Zhang, Dongxiao; Peng, Nan; Zhao, Shumiao; Liang, Yunxiang

    2015-01-01

    Lactic acid bacteria (LAB) have been shown to enhance performance of weaned piglets. However, few studies have reported the addition of LAB Enterococcus faecalis as alternatives to growth promoting antibiotics for weaned piglets. This study evaluated the effects of dietary E. faecalis LAB31 on the growth performance, diarrhea incidence, blood parameters, fecal bacterial and Lactobacillus communities in weaned piglets. A total of 360 piglets weaned at 26 ± 2 days of age were randomly allotted to 5 groups (20 pens, with 4 pens for each group) for a trial of 28 days: group N (negative control, without antibiotics or probiotics); group P (Neomycin sulfate, 100 mg/kg feed); groups L, M and H (supplemented with E. faecalis LAB31 0.5×109, 1.0×109, and 2.5×109 CFU/kg feed, respectively). Average daily gain and feed conversion efficiency were found to be higher in group H than in group N, and showed significant differences between group H and group P (P0 < 0.05). Furthermore, groups H and P had a lower diarrhea index than the other three groups (P0 < 0.05). Denaturing gradient gel electrophoresis (DGGE) showed that the application of probiotics to the diet changed the bacterial community, with a higher bacterial diversity in group M than in the other four groups. Real-time PCR revealed that the relative number of Lactobacillus increased by addition of probiotics, and was higher in group H than in group N (P0 < 0.05). However, group-specific PCR-DGGE showed no obvious difference among the five groups in Lactobacillus composition and diversity. Therefore, the dietary addition of E. faecalis LAB31 can improve growth performance, reduce diarrhea, and increase the relative number of Lactobacillus in feces of weaned piglets. PMID:25617897

  7. [Effect of MDG-1, a polysaccharide from Ophiopogon japonicas, on diversity of lactobacillus in diet-induced obese mice].

    PubMed

    Shi, Lin-lin; Wang, Yuan; Feng, Yi

    2015-02-01

    For understanding the effect of MDG-1, a water-soluble ?-D-fructan polysaccharide from Ophiopogon japonicas, on intestinal microecological balance, especially on the changes of lactobacillus, sixty 8-week-old male C57BL/6J mice were given a high-fat diet for six weeks and were also gavaged with saline once a day simultaneously. Then the mice which is below 30 grams or dropped more than 10% through lavage were eliminated and the rest were randomly divided into four groups: diet-induced obese (DIO) model group (n = 12, gavaged with saline), low-dose MDG-1 group (n = 12, gavaged with MDG-1, 75 mg · kg(-1)) , medial-dose MDG- 1 group (n = 12, gavaged with 150 mg · kg(-1)), and high-dose MDG-1 group (n = 12, gavaged with 300 mg · kg(-1)) according to the weight and blood glucose; the model group and MDG-1 group were placed on a high-fat diet while the normal control group (n = 12, gavaged with saline) were kept on a low-fat diet through the experiment. After 12-weeks of treatment, feces samples were collected and cultured for intestinal microecological balance analysis. Then the intestinal probiotics were cultured through traditional methods combined with modified gradient gel electrophoresis (DGGE) method. The changes of lactobacillus in each treatment group were also detected by a statistical analysis of the total number of the intestinal flora. We have established the phylogenetic tree by 16S rDNA sequencing and use some molecular identification methods such as PCR-DGGE to analyse the changes of the dominant bacteria floras, and also get the pure culture. In conclusion, different concentrations of MDG-1 can increase the number of the intestinal probiotics, especially Taiwan lactobacillus and Lactobacillus murinus, and improve their diversity and promote proliferation in a dose-dependent way. PMID:26137696

  8. Complete genome sequence of Lactobacillus paracasei CAUH35, a new strain isolated from traditional fermented dairy product koumiss in China.

    PubMed

    Wang, Guohong; Xiong, Yao; Xu, Qi; Yin, Jia; Hao, Yanling

    2015-11-20

    Lactobacillus paracasei CAUH35 was isolated from homemade koumiss, a traditional fermented dairy product with beneficial effects on human health. The genome consists of a circular 2,770,411bp chromosome and four plasmids. Genome analysis revealed the presence of gene clusters involved in the production of exopolysaccharides and bacteriocin. The complete genome sequence of L. paracasei CAUH35 will provide genetic basis for further comparative and functional genomic analyses. PMID:26393953

  9. In vitro probiotic characteristics of Lactobacillus plantarum ZDY 2013 and its modulatory effect on gut microbiota of mice.

    PubMed

    Huang, Renhui; Tao, Xueying; Wan, Cuixiang; Li, Shengjie; Xu, Hengyi; Xu, Feng; Shah, Nagendra P; Wei, Hua

    2015-09-01

    Lactobacillus plantarum ZDY 2013, a novel strain isolated from Chinese traditional fermented acid beans, was systematically evaluated for its survival capacity under stress conditions (pH, bile salt, simulated gastrointestinal tract, and antibiotics), production of exopolysaccharide and antagonism against 8 pathogens. Its effect on mice gut microbiota was also investigated by quantitative PCR and PCR-denaturing gradient gel electrophoresis. The results showed that ZDY 2013 can grow at pH 3.5 and survive at pH 2.0 for 6 h and at 0.45% bile salt for 3 h. The exopolysaccharide yield was up to 204±7.68 mg/L. The survival rate of ZDY 2013 in a simulated gastrointestinal tract was as high as 65.84%. Antagonism test with a supernatant of ZDY 2013 showed maximum halo of 28 mm against Listeria monocytogenes. The inhibition order was as follows: Listeria monocytogenes, Salmonella typhimurium, Escherichia coli, Pseudomonas aeruginosa, Shigella sonnei, Enterobacter sakazakii, and Staphylococcus aureus. Lactobacillus plantarum ZDY 2013 was sensitive to some antibiotics (e.g., macrolide, sulfonamides, aminoglycoside, tetracyclines and ?-lactams), whereas it was resistant to glycopeptides, quinolones, and cephalosporins antibiotics. Denaturing gradient gel electrophoresis profile demonstrated that ZDY 2013 administration altered the composition of the microbiota at various intestinal loci of the mice. Moreover, the quantitative PCR test showed that the administration of ZDY 2013 enhanced the populations of Bifidobacterium and Lactobacillus in either the colon or cecum, and reduced the potential enteropathogenic bacteria (e.g., Enterococcus, Enterobacterium, and Clostridium perfringens). Lactobacillus plantarum ZDY 2013 exhibited high resistance against low pH, bile salt, and gastrointestinal fluid, and possessed antibacterial and gut microbiota modulation properties with a potential application in the development of dairy food and nutraceuticals. PMID:26142853

  10. A novel biosensor based on Lactobacillus acidophilus for determination of phenolic compounds in milk products and wastewater.

    PubMed

    Sagiroglu, Ayten; Paluzar, Hatice; Ozcan, Hakki Mevlut; Okten, Suzan; Sen, Burhan

    2011-01-01

    Different branches of industry need to use phenolic compounds (PCs) in their production, so determination of PCs sensitively, accurately, rapidly, and economically is very important. For the sensitive determination of PCs, some biosensors based on pure polyphenol oxidase, plant tissue and microorganisms were developed before. But there has been no study to develop a microbial phenolic compounds biosensor based on Lactobacillus species, which contain polyphenol oxidase enzyme. In this study, we used different forms of Lactobacillus species as enzyme sources of biosensor and compared biosensor performances of these forms for determination of PCs. For this purpose, we used lyophilized Lactobacillus cells (containing L. bulgaricus, L. acidophilus, Streptococcus thermophilus), pure L. acidophilus, pure L. bulgaricus, and L. acidophilus- and L. bulgaricus adapted to catechol in Lactobacilli MRS Broth. The most suitable form was determined and optimization studies of the biosensor were carried out by using this form. For preparing the bioactive layer of the biosensor, the Lactobacillus cells were immobilized in gelatin by using glutaraldehyde. In the study, we used catechol as a substrate. Phenolic compound determination is based on the assay of the differences on the respiration activity of the cells on the oxygen meter in the absence and the presence of catechol. The microbial biosensor response depends directly on catechol concentration between 0.5 and 5.0 mM with 18 min response time. In the optimization studies of the microbial biosensor the most suitable microorganism amount was found to be 10 mg, and also phosphate buffer (pH 8.0; 50 mM) and 37.5 °C were obtained as the optimum working conditions. In the characterization studies of the microbial biosensor some parameters such as substrate specificity on the biosensor response and operational and storage stability were examine. Furthermore, the determination of PC levels in synthetic wastewater, industrial wastewater, and milk products was investigated by using the developed biosensor under optimum conditions. PMID:21967334

  11. Effect of Lactobacillus johnsonii CRL1647 on different parameters of honeybee colonies and bacterial populations of the bee gut.

    PubMed

    Audisio, M C; Sabaté, D C; Benítez-Ahrendts, M R

    2015-10-15

    Lactobacillus johnsonii CRL1647, isolated from the intestinal tract of a worker-bee in Salta, Argentina, was delivered to Apis mellifera L. honey bee colonies according to two different administration schedules: 1×10(5) cfu/ml every 15 days (2011) or monthly (2012). The effect of each treatment on the bee-colony performance was monitored by measuring honey production, and the prevalence of varroasis and nosemosis. Worker bees from each assay were randomly captured 3 days after administration and assayed for the following intestinal culturable and defined bacterial populations: total aerobic microorganisms, Bacillus spp. spores, Lactobacillus spp., Enterococcus spp. and enterobacteria. Interestingly, both treatments generated a similar increase in honey production in treated colonies compared to controls: 36.8% (every 15 days) and 36.3% (monthly). Nosema index always exhibited a reduction when lactobacilli were administered; in turn, Varroa incidence was lower when the lactobacilli were administered once a month. Moreover, the administration of L. johnsonii CRL1647 every 15 days produced an increase in the total number of aerobic microorganisms and in bacteria belonging to the genera Lactobacillus and Enterococcus; at the same time, a decrease was observed in the number of total spores at the end of the treatment. The number of enterobacteria was constant and remained below that of control hives at the end of the assay. On the other hand, the delivery of lactobacilli once a month only showed an increase in the number of bacteria belonging to the genus Lactobacillus; meanwhile, viable counts of the remaining microorganisms assayed were reduced. Even though it seems that both treatments were similar, those bee colonies that received L. johnsonii CRL1647 every 15 days became so strong that they swarmed. PMID:25809216

  12. Immunomodulatory activity and control of Salmonella Enteritidis colonization in the intestinal tract of chickens by Lactobacillus based probiotic.

    PubMed

    Penha Filho, Rafael Antonio Casarin; Díaz, Silvia Juliana Acelas; Fernando, Filipe Santos; Chang, Yung-Fu; Andreatti Filho, Raphael Lucio; Berchieri Junior, Angelo

    2015-09-15

    Lactobacillus-based probiotics (LBP) are used as competitive exclusion to control pathogenic enterobacterial infections and improve the weight gain in broiler chickens. This study assessed the inhibition of Salmonella Enteritidis (SE) infection in one-week-old broiler chicks, using an experimental LBP containing four Lactobacillus strains isolated from chickens (L. acidophilus, L. fermentum, L. reuteri, L. salivarius). The immunomodulatory effects of this treatment were evaluated, through the analysis of cytokines and influx of macrophages, ??, CD4(+) and CD8(+) T cells in the gut. The intestinal colonization by SE was reduced by 1.8 CFU/g (log10) in chicks treated with LBP (p<0.05). The levels of pro-inflammatory cytokines (IL-1?, LITAF) were significantly reduced in treated chicks (p<0.05), whilst untreated chicks showed elevated inflammatory stimulus and an increased population of CD8(+) T cells in the intestinal mucosa after challenge (p<0.05). Additionally, the LBP stimulated TLR2 expression in caecal tonsils. The adjuvant property of the Lactobacillus cell wall (LCW) was evaluated, demonstrating good capability to stimulate T helper 2 (Th2) cell proliferation. Pretreatment of chicks with LBP decreased the intestinal colonization by SE, minimizing the tissue lesions and inflammation after challenge and showed a potential use as adjuvant with injectable killed vaccines. PMID:26099807

  13. Isolation, Identification and Partial Characterization of a Lactobacillus casei Strain with Bile Salt Hydrolase Activity from Pulque.

    PubMed

    González-Vázquez, R; Azaola-Espinosa, A; Mayorga-Reyes, L; Reyes-Nava, L A; Shah, N P; Rivera-Espinoza, Y

    2015-12-01

    The aim of this study was to isolate, from pulque, Lactobacillus spp. capable of survival in simulated gastrointestinal stress conditions. Nine Gram-positive rods were isolated; however, only one strain (J57) shared identity with Lactobacillus and was registered as Lactobacillus casei J57 (GenBank accession: JN182264). The other strains were identified as Bacillus spp. The most significant observation during the test of tolerance to simulated gastrointestinal conditions (acidity, gastric juice and bile salts) was that L. casei J57 showed a rapid decrease (p ? 0.05) in the viable population at 0 h. Bile salts were the stress condition that most affected its survival, from which deoxycholic acid and the mix of bile salts (oxgall) were the most toxic. L. casei J57 showed bile salt hydrolase activity over primary and secondary bile salts as follows: 44.91, 671.72, 45.27 and 61.57 U/mg to glycocholate, taurocholate, glycodeoxycholate and taurodeoxycholate. In contrast, the control strain (L. casei Shirota) only showed activity over tauroconjugates. These results suggest that L. casei J57 shows potential for probiotic applications. PMID:26566892

  14. Enhanced Trapping of HIV-1 by Human Cervicovaginal Mucus Is Associated with Lactobacillus crispatus-Dominant Microbiota

    PubMed Central

    Nunn, Kenetta L.; Wang, Ying-Ying; Harit, Dimple; Humphrys, Michael S.; Ma, Bing; Cone, Richard; Ravel, Jacques

    2015-01-01

    ABSTRACT Cervicovaginal mucus (CVM) can provide a barrier that precludes HIV and other sexually transmitted virions from reaching target cells in the vaginal epithelium, thereby preventing or reducing infections. However, the barrier properties of CVM differ from woman to woman, and the causes of these variations are not yet well understood. Using high-resolution particle tracking of fluorescent HIV-1 pseudoviruses, we found that neither pH nor Nugent scores nor total lactic acid levels correlated significantly with virus trapping in unmodified CVM from diverse donors. Surprisingly, HIV-1 was generally trapped in CVM with relatively high concentrations of d-lactic acid and a Lactobacillus crispatus-dominant microbiota. In contrast, a substantial fraction of HIV-1 virions diffused rapidly through CVM with low concentrations of d-lactic acid that had a Lactobacillus iners-dominant microbiota or significant amounts of Gardnerella vaginalis, a bacterium associated with bacterial vaginosis. Our results demonstrate that the vaginal microbiota, including specific species of Lactobacillus, can alter the diffusional barrier properties of CVM against HIV and likely other sexually transmitted viruses and that these microbiota-associated changes may account in part for the elevated risks of HIV acquisition linked to bacterial vaginosis or intermediate vaginal microbiota. PMID:26443453

  15. The TRPV1 channel in rodents is a major target for antinociceptive effect of the probiotic Lactobacillus reuteri DSM 17938.

    PubMed

    Perez-Burgos, Azucena; Wang, Lu; McVey Neufeld, Karen-Anne; Mao, Yu-Kang; Ahmadzai, Mustafa; Janssen, Luke J; Stanisz, Andrew M; Bienenstock, John; Kunze, Wolfgang A

    2015-09-01

    Certain probiotic bacteria have been shown to reduce distension-dependent gut pain, but the mechanisms involved remain obscure. Live luminal Lactobacillus reuteri (DSM 17938) and its conditioned medium dose dependently reduced jejunal spinal nerve firing evoked by distension or capsaicin, and 80% of this response was blocked by a specific TRPV1 channel antagonist or in TRPV1 knockout mice. The specificity of DSM action on TRPV1 was further confirmed by its inhibition of capsaicin-induced intracellular calcium increases in dorsal root ganglion neurons. Another lactobacillus with ability to reduce gut pain did not modify this response. Prior feeding of rats with DSM inhibited the bradycardia induced by painful gastric distension. These results offer a system for the screening of new and improved candidate bacteria that may be useful as novel therapeutic adjuncts in gut pain. Certain bacteria exert visceral antinociceptive activity, but the mechanisms involved are not determined. Lactobacillus reuteri DSM 17938 was examined since it may be antinociceptive in children. Since transient receptor potential vanilloid 1 (TRPV1) channel activity may mediate nociceptive signals, we hypothesized that TRPV1 current is inhibited by DSM. We tested this by examining the effect of DSM on the firing frequency of spinal nerve fibres in murine jejunal mesenteric nerve bundles following serosal application of capsaicin. We also measured the effects of DSM on capsaicin-evoked increase in intracellular Ca(2+) or ionic current in dorsal root ganglion (DRG) neurons. Furthermore, we tested the in vivo antinociceptive effects of oral DSM on gastric distension in rats. Live DSM reduced the response of capsaicin- and distension-evoked firing of spinal nerve action potentials (238 ± 27.5% vs. 129 ± 17%). DSM also reduced the capsaicin-evoked TRPV1 ionic current in DRG neuronal primary culture from 83 ± 11% to 41 ± 8% of the initial response to capsaicin only. Another lactobacillus (Lactobacillus rhamnosus JB-1) with known visceral anti-nociceptive activity did not have these effects. DSM also inhibited capsaicin-evoked Ca(2+) increase in DRG neurons; an increase in Ca(2+) fluorescence intensity ratio of 2.36 ± 0.31 evoked by capsaicin was reduced to 1.25 ± 0.04. DSM releasable products (conditioned medium) mimicked DSM inhibition of capsaicin-evoked excitability. The TRPV1 antagonist 6-iodonordihydrocapsaicin or the use of TRPV1 knock-out mice revealed that TRPV1 channels mediate about 80% of the inhibitory effect of DSM on mesenteric nerve response to high intensity gut distension. Finally, feeding with DSM inhibited perception in rats of painful gastric distension. Our results identify a specific target channel for a probiotic with potential therapeutic properties. PMID:26084409

  16. Role of Lactobacillus Species in the Intermediate Vaginal Flora in Early Pregnancy: A Retrospective Cohort Study

    PubMed Central

    Farr, Alex; Kiss, Herbert; Hagmann, Michael; Machal, Susanne; Holzer, Iris; Kueronya, Verena; Husslein, Peter Wolf; Petricevic, Ljubomir

    2015-01-01

    Background Poor obstetrical outcomes are associated with imbalances in the vaginal flora. The present study evaluated the role of vaginal Lactobacillus species in women with intermediate vaginal flora with regard to obstetrical outcomes. Methods We retrospectively analysed data from all women with singleton pregnancies who had undergone routine screening for asymptomatic vaginal infections at our tertiary referral centre between 2005 and 2014. Vaginal smears were Gram-stained and classified according to the Nugent scoring system as normal flora (score 0–3), intermediate vaginal flora (4–6), or bacterial vaginosis (7–10). Only women with intermediate vaginal flora were investigated. Women with a Nugent score of 4 were categorised into those with and without Lactobacilli. Follow-up smears were obtained 4–6 weeks after the initial smears. Descriptive data analysis, the Welch’s t-test, the Fisher’s exact test, and multiple regression analysis with adjustment for confounders were performed. Gestational age at delivery and birth weight were the outcome measures. Results At antenatal screening, 529/8421 women presented with intermediate vaginal flora. Amongst these, 349/529 (66%) had a Nugent score of 4, 94/529 (17.8%) a Nugent score of 5, and 86/529 (16.2%) a Nugent score of 6. Amongst those with a Nugent score of 4, 232/349 (66.5%) women were in the Lactobacilli group and 117/349 (33.5%) in the Non-Lactobacilli group. The preterm delivery rate was significantly lower in the Lactobacilli than in the Non-Lactobacilli group (OR 0.34, CI 0.21–0.55; p<0.001). Mean birth weight was 2979 ± 842 g and 2388 ± 1155 g in the study groups, respectively (MD 564.12, CI 346.23–781.92; p<0.001). On follow-up smears, bacterial vaginosis rates were 9% in the Lactobacilli and 7.8% in the Non-Lactobacilli group. Conclusions The absence of vaginal Lactobacillus species and any bacterial colonisation increases the risks of preterm delivery and low birth weight in women with intermediate vaginal flora in early pregnancy. PMID:26658473

  17. Examination of the technological properties of newly isolated strains of the genus Lactobacillus and possibilities for their application in the composition of starters

    PubMed Central

    Denkova, Rositsa; Ilieva, Svetla; Denkova, Zapryana; Georgieva, Ljubka; Krastanov, Albert

    2014-01-01

    The ability of four Lactobacillus strains – Lactobacillus brevis LBRZ7 (isolated from fermented cabbage), Lactobacillus plantarum LBRZ12 (isolated from fermented cabbage), Lactobacillus fermentum LBRH9 (of human origin) and Lactobacillus casei ssp. rhamnosus LBRC11 (isolated from home-made cheese) – to grow in flour/water environment and to accumulate high concentrations of viable cells was examined. Two starters for sourdough were created for lab-scale production of wheat bread: a two-strain starter and a four-strain starter. Wheat bread with improved properties – greater loaf volume, enhanced flavour and softer and brighter crumb – was obtained from the 7% four-strain starter sourdough. The addition of sourdough in the production of wheat bread affected positively the technological and organoleptic characteristics of the final bread by inhibiting the growth of wild yeasts and mold and Bacillus spores without the addition of preservatives. The inclusion of 15% of the four-strain starter sourdough in the bread-making process led to enhanced safety and longer shelf life of the baked bread. PMID:26019534

  18. Identification and characterization of a glucan-producing enzyme from Lactobacillus hilgardii TMW 1.828 involved in granule formation of water kefir.

    PubMed

    Waldherr, Florian W; Doll, Viktoria M; Meissner, Daniel; Vogel, Rudi F

    2010-08-01

    Water kefir is a home made fermented beverage based on a sucrose solution with fruit extracts. The inoculum of such fermentations consists of macroscopic granula containing lactic and acetic acid bacteria, and yeasts, which are embedded in an exopolysaccharide (EPS) matrix. In this work, a strain of Lactobacillus hilgardii producing large amounts of the granule-forming dextran could be isolated. The glycosyltransferans (Gtf) commonly called glucansucrase responsible for the production of this dextran was purified from L. hilgardii. Characteristic enzyme kinetic data were obtained. Optimum activity was observed between pH 4.3 and 4.6 and temperatures between 40 degrees C and 45 degrees C. A Michaelis-Menten kinetic could be fit to the experimental data and a K(M) of 0.0385 M was calculated. The corresponding gtf gene was identified and characterized. It encodes a 1448 amino acid protein with higher homologies to Gtfs of Lactobacillus parabuchneri, Lactobacillus sakei and Lactobacillus fermentum followed by lower homologies to Lactobacillus reuteri Gtfs. By knockout experiments the role of this gene in granule dextran production was demonstrated. PMID:20510787

  19. Improving Properties of a Novel ?-Galactosidase from Lactobacillus plantarum by Covalent Immobilization.

    PubMed

    Benavente, Rocio; Pessela, Benevides C; Curiel, Jose Antonio; de las Rivas, Blanca; Muñoz, Rosario; Guisán, Jose Manuel; Mancheño, Jose M; Cardelle-Cobas, Alejandra; Ruiz-Matute, Ana I; Corzo, Nieves

    2015-01-01

    A novel ?-galactosidase from Lactobacillus plantarum (LPG) was over-expressed in E. coli and purified via a single chromatographic step by using lowly activated IMAC (immobilized metal for affinity chromatography) supports. The pure enzyme exhibited a high hydrolytic activity of 491 IU/mL towards o-nitrophenyl ?-D-galactopyranoside. This value was conserved in the presence of different divalent cations and was quite resistant to the inhibition effects of different carbohydrates. The pure multimeric enzyme was stabilized by multipoint and multisubunit covalent attachment on glyoxyl-agarose. The glyoxyl-LPG immobilized preparation was over 20-fold more stable than the soluble enzyme or the one-point CNBr-LPG immobilized preparation at 50 °C. This ?-galactosidase was successfully used in the hydrolysis of lactose and lactulose and formation of different oligosaccharides was detected. High production of galacto-oligosaccharides (35%) and oligosaccharides derived from lactulose (30%) was found and, for the first time, a new oligosaccharide derived from lactulose, tentatively identified as 3'-galactosyl lactulose, has been described. PMID:25942370

  20. Lipoteichoic Acid Isolated from Lactobacillus plantarum Inhibits Melanogenesis in B16F10 Mouse Melanoma Cells

    PubMed Central

    Kim, Hye Rim; Kim, Hangeun; Jung, Bong Jun; You, Ga Eun; Jang, Soojin; Chung, Dae Kyun

    2015-01-01

    Lipoteichoic acid (LTA) is a major component of the cell wall of Gram-positive bacteria. Its effects on living organisms are different from those of lipopolysaccharide (LPS) found in Gram-negative bacteria. LTA contributes to immune regulatory effects including anti-aging. In this study, we showed that LTA isolated from Lactobacillus plantarum (pLTA) inhibited melanogenesis in B16F10 mouse melanoma cells. pLTA reduced the cellular activity of tyrosinase and the expression of tyrosinase family members in a dose-dependent manner. The expression of microphthalmia-associated transcription factor (MITF), a key factor in the synthesis of melanin, was also decreased by pLTA. Further, we showed that pLTA activated melanogenesis signaling, such as extracellular signal-regulated kinase (ERK) and phosphatidylinositol 3-kinse (PI3K)/AKT. In addition, the expression of heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) and HuR, which are important RNA-binding proteins (RBPs), was reduced. pLTA likely degrades MITF via regulation of melanogenic signaling and RNA stability of melanogenic proteins, resulting in the reduction of melanin. Thus, our data suggest that pLTA has therapeutic potential for treating hyperpigmentation disorders and can also be used as a cosmetic whitening agent. PMID:26021887

  1. Lactobacillus brevis Strains from Fermented Aloe vera Survive Gastroduodenal Environment and Suppress Common Food Borne Enteropathogens

    PubMed Central

    Kim, Young-Wook; Jeong, Young-Ju; Kim, Ah-Young; Son, Hyun-Hee; Lee, Jong-Am; Jung, Cheong-Hwan; Kim, Chae-Hyun; Kim, Jaeman

    2014-01-01

    Five novel Lactobacillus brevis strains were isolated from naturally fermented Aloe vera leaf flesh. Each strain was identified by Random Amplified Polymorphic DNA (RAPD) analysis and 16S rRNA sequence comparison. These strains were highly tolerant to acid, surviving in pH2.5 for up to 4 hours, and resistant to 5% bile salts at 37°C for 18 hours. Due to its tolerance to acid and bile salts, one strain passed through the gastric barrier and colonised the intestine after oral administration. All five strains inhibited the growth of many harmful enteropathogens without restraining most of normal commensals in the gut and hence named POAL (Probiotics Originating from Aloe Leaf) strains. Additionally, each strain exhibited discriminative resistance to a wide range of antibiotics. The L. brevis POAL strains, moreover, expressed high levels of the glutamate decarboxylase (GAD) gene which produces a beneficial neurotransmitter, ?-aminobutyric acid (GABA). These characteristics in all suggest that the novel L. brevis strains should be considered as potential food additives and resources for pharmaceutical research. PMID:24598940

  2. Diversity of Lactobacillus reuteri Strains in Converting Glycerol into 3-Hydroxypropionic Acid.

    PubMed

    Burgé, G; Saulou-Bérion, C; Moussa, M; Pollet, B; Flourat, A; Allais, F; Athès, V; Spinnler, H E

    2015-10-01

    The present study aims at comparing the performances of three Lactobacillus reuteri strains (DSM 20016, DSM 17938, and ATCC 53608) in producing 3-hydroxypropionic acid (3-HP) from glycerol and at exploring inhibition phenomena during this bioconversion. Differences were highlighted between the three strains in terms of 3-HP production yield, kinetics of substrate consumption, and metabolite production. With a maximal productivity in non-optimal conditions (free pH) around 2 g.L(-1).h(-1) of 3-HP and 4 g.L(-1).h(-1) of 3-hydroxypropionaldehyde (3-HPA) depending on the strain, this study confirmed the potential of L. reuteri for the biotechnological production of 3-HP. Moreover, the molar ratios of 3-HP to 1,3-propanediol (1,3-PDO) obtained for the three strains (comprised between 1.25 and 1.65) showed systematically a higher 3-HP production. From these results, the DSM 17938 strain appeared to be the most promising strain. The impact of glycerol bioconversion on the bacteria's physiological state (a decrease of around 40 % in DSM 17938 cells showing an enzymatic activity after 3 h) and survival (total loss of cultivability after 2 or 3 h depending on the strains) was revealed and discussed. The effect of each metabolite on L. reuteri DSM 17938 was further investigated, displaying a drastic inhibition caused by 3-HPA, while 3-HP induced lower impact and only at acidic pH. PMID:26319567

  3. Bio-transformation of Glycerol to 3-Hydroxypropionic Acid Using Resting Cells of Lactobacillus reuteri.

    PubMed

    Ramakrishnan, Gopi Gopal; Nehru, Ganesh; Suppuram, Pandiaraj; Balasubramaniyam, Sowmiya; Gulab, Brajesh Raman; Subramanian, Ramalingam

    2015-10-01

    Lactobacillus reuteri grown in MRS broth containing 20 mM glycerol exhibits 3.7-fold up-regulation of 3-hydroxypropionic acid (3-HP) pathway genes during the stationary phase. Concomitantly, the resting cells prepared from stationary phase show enhancement in bio-conversion of glycerol, and the maximum specific productivity (q p) is found to be 0.17 g 3-HP per g CDW per hour. The regulatory elements such as catabolite repression site in the up-stream of 3-HP pathway genes are presumed for the augmentation of glycerol bio-conversion selectively in stationary phase. However, in the repression mutant, the maximum q p of 3-HP persisted in the stationary phase-derived resting cells indicating the role of further regulatory features. In the production stage, the external 3-HP concentration of 35 mM inhibits 3-HP synthesis. In addition, it has also moderated 1,3-propanediol formation, as it is a redox bio-catalysis involving NAD(+)/NADH ratio of 6.5. Repeated batch bio-transformation has been used to overcome product inhibition, and the total yield (Ypx) of 3-HP from the stationary phase-derived biomass is 3.3 times higher than that from the non-repeated mode. With the use of appropriate gene expression condition and repeated transfer of biomass, 3-HP produced in this study can be used for low-volume, high-value applications. PMID:26204968

  4. Effect of immobilized Lactobacillus casei on volatile compounds of heat treated probiotic dry-fermented sausages.

    PubMed

    Sidira, Marianthi; Kandylis, Panagiotis; Kanellaki, Maria; Kourkoutas, Yiannis

    2015-07-01

    The effect of the amount of immobilized Lactobacillus casei ATCC 393 on wheat grains on the generation of volatile compounds during the production of heat treated probiotic dry-fermented sausages was investigated. For comparison reasons, sausages containing free L. casei cells or no starter culture as well as a similar commercial product were also included in the study. Samples ripened for 8 days and heat treated to 70-72°C for 8-10 min were subjected to Solid Phase Microextraction (SPME) Gas Chromatography/Mass Spectrometry (GC/MS) analysis. The starter culture affected significantly the production of volatile compounds. The highest content of esters and alcohols was observed in the sample containing 30 g of immobilized cells/kg of stuffing mixture, while the highest concentration of organic acids was observed in the sausages with no starter culture. In contrast, the commercial product contained the lowest concentration of volatiles. Principal component analysis of the semi-quantitative data revealed that the volatile composition was affected primarily by the nature and concentration of the starter culture. PMID:25704702

  5. Characterization and molecular cloning of cryptic plasmids isolated from Lactobacillus casei.

    PubMed Central

    Lee-Wickner, L J; Chassy, B M

    1985-01-01

    Four small cryptic plasmids were isolated from Lactobacillus casei strains, and restriction endonuclease maps of these plasmids were constructed. Three of the small plasmids (pLZ18C, pLZ19E, and pLZ19F1; 6.4, 4.9, and 4.8 kilobase pairs, respectively) were cloned into Escherichia coli K-12 by using pBR322, pACYC184, and pUC8 as vectors. Two of the plasmids, pLZ18C and pLZ19E, were also cloned into Streptococcus sanguis by using pVA1 as the vector. Hybridization by using nick-translated cloned 32P-labeled L. casei plasmid DNA as the probe revealed that none of the cryptic plasmids had appreciable DNA-DNA homology with the large lactose plasmids found in the L. casei strains, with chromosomal DNAs isolated from these strains. Partial homology was detected among several plasmids isolated from different strains, but not among cryptic plasmids isolated from the same strain. Images PMID:3923929

  6. Competitive Growth of Genetically Marked Malolactic-Deficient Lactobacillus plantarum in Cucumber Fermentations

    PubMed Central

    Breidt, Fred; Fleming, Henry P.

    1992-01-01

    Procedures were developed for the differential enumeration of an added strain of Lactobacillus plantarum and indigenous lactic acid bacteria (LAB) during the fermentation of brined cucumbers. The added strain was an N,N-nitrosoguanidine-generated mutant that lacked the ability to produce CO2 from malic acid (MDC-). The MDC- phenotype is desirable because CO2 production from malic acid decarboxylation has been shown to contribute to bloater formation in fermented cucumbers. A basal medium containing malic acid and adjusted to pH 4.0 permitted growth of indigenous LAB (predominantly MDC+), but not growth of the added MDC- culture. Transformation of the MDC- culture by electroporation with cloning vector pGK12 conferred chloramphenicol resistance, which permitted selective enumeration of this culture. The reversion frequency of the MDC- mutation was determined by a fluctuation test to be less than 10-10. The level of retention of plasmid pGK12 was greater than 90% after 10 generations in cucumber juice medium at 32°C. With the procedures developed, we were able to establish the ratio of MDC- to MDC+ LAB that results in malic acid retention in fermentations of filter-sterilized cucumber juice and unsterilized whole cucumbers under specified conditions. PMID:16348818

  7. Development of microparticulate systems for intestinal delivery of Lactobacillus acidophilus and Bifidobacterium lactis.

    PubMed

    Albertini, Beatrice; Vitali, Beatrice; Passerini, Nadia; Cruciani, Federica; Di Sabatino, Marcello; Rodriguez, Lorenzo; Brigidi, Patrizia

    2010-07-11

    In the present study intestinal delivery systems resistant to gastric juice, loaded with the probiotic bacteria Lactobacillus acidophilus LA14 and Bifidobacterium lactis BI07, were produced by the polyelectrolyte complexation. First, beads were prepared by the traditional extrusion method and nine formulations were developed using alginate as main carrier and the biopolymer, xanthan gum (XG), as hydrophilic retardant polymer or the cellulose derivative, cellulose acetate phthalate (CAP), as gastro-resistant polymer. The results showed that the incorporation of the 0.5% (w/v) of XG or the 1% (w/v) of CAP within the 3% (w/v) of alginate solution increased the survival of the probiotic bacteria in acid conditions from 63% of the freeze-dried bacteria up to 76%. Subsequently, these formula was used to prepare smaller microcapsules by means of an atomization device. Despite of the high viscosity of the biomass suspension, the spraying system produced spherical and non-aggregated microcapsules able to survive in harsh condition better than beads: the survival of the probiotic bacteria after acid incubation was 91%. The performance of the microcapsules in simulated gastric fluid (SGF) containing pepsin and in gut medium (GM) containing bile salts was excellent (viability>95%). Furthermore, the viability of probiotic bacteria was maintained after an incubation of 24h in GM. Finally, stability tests performed at 5 degrees C highlighted a bacterial viability of about 82% and 70% after 6 and 9 months, respectively. PMID:20420903

  8. Metabolic footprinting of Lactobacillus buchneri strain LA1147 during anaerobic spoilage of fermented cucumbers.

    PubMed

    Johanningsmeier, Suzanne D; McFeeters, Roger F

    2015-12-23

    Lactobacillus buchneri has recently been associated with anaerobic spoilage of fermented cucumbers due to its ability to metabolize lactic acid into acetic acid and 1,2-propanediol. However, we have limited knowledge of other chemical components in fermented cucumber that may be related to spoilage and the unique metabolic capabilities of L. buchneri. Comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry metabolite profiling methods were applied for nontargeted detection of volatile and nonvolatile compounds to determine changes that occurred during anaerobic fermented cucumber spoilage by L. buchneri LA1147 and during reproduction of spoilage with natural microbiota. Univariate analysis of variance combined with hierarchial clustering analysis revealed 92 metabolites that changed during spoilage (P<0.01). Decreases were observed in mono and disaccharides, amino acids, nucleosides, long chain fatty acids, aldehydes, and ketones, and increases were observed in several alcohols and butanoic and pentanoic acids. Most of the metabolite changes preceded lactic acid utilization, indicating that lactic acid is not a preferred substrate for anaerobic spoilage organisms in fermented cucumbers. The ability to detect biochemical changes that preceded lactate utilization revealed citrulline, trehalose, and cellobiose as compounds that may signify metabolic activity of L. buchneri spoilage strains prior to any significant product degradation. PMID:26325599

  9. Impact of Environmental Factors on Bacteriocin Promoter Activity in Gut-Derived Lactobacillus salivarius.

    PubMed

    Guinane, Caitriona M; Piper, Clare; Draper, Lorraine A; O'Connor, Paula M; Hill, Colin; Ross, R Paul; Cotter, Paul D

    2015-11-15

    Bacteriocin production is regarded as a desirable probiotic trait that aids in colonization and persistence in the gastrointestinal tract (GIT). Strains of Lactobacillus salivarius, a species associated with the GIT, are regarded as promising probiotic candidates and have a number of associated bacteriocins documented to date. These include multiple class IIb bacteriocins (salivaricin T, salivaricin P, and ABP-118) and the class IId bacteriocin bactofencin A, which show activity against medically important pathogens. However, the production of a bacteriocin in laboratory media does not ensure production under stressful environmental conditions, such as those encountered within the GIT. To allow this issue to be addressed, the promoter regions located upstream of the structural genes encoding the L. salivarius bacteriocins mentioned above were fused to a number of reporter proteins (green fluorescent protein [GFP], red fluorescent protein [RFP], and luciferase [Lux]). Of these, only transcriptional fusions to GFP generated signals of sufficient strength to enable the study of promoter activity in L. salivarius. While analysis of the class IIb bacteriocin promoter regions indicated relatively weak GFP expression, assessment of the promoter of the antistaphylococcal bacteriocin bactofencin A revealed a strong promoter that is most active in the absence of the antimicrobial peptide and is positively induced in the presence of mild environmental stresses, including simulated gastric fluid. Taken together, these data provide information on factors that influence bacteriocin production, which will assist in the development of strategies to optimize in vivo and in vitro production of these antimicrobials. PMID:26341205

  10. Histamine-Producing Pathway Encoded on an Unstable Plasmid in Lactobacillus hilgardii 0006

    PubMed Central

    Lucas, Patrick M.; Wolken, Wout A. M.; Claisse, Olivier; Lolkema, Juke S.; Lonvaud-Funel, Aline

    2005-01-01

    Histamine production from histidine in fermented food products by lactic acid bacteria results in food spoilage and is harmful to consumers. We have isolated a histamine-producing lactic acid bacterium, Lactobacillus hilgardii strain IOEB 0006, which could retain or lose the ability to produce histamine depending on culture conditions. The hdcA gene, coding for the histidine decarboxylase of L. hilgardii IOEB 0006, was located on an 80-kb plasmid that proved to be unstable. Sequencing of the hdcA locus disclosed a four-gene cluster encoding the histidine decarboxylase, a protein of unknown function, a histidyl-tRNA synthetase, and a protein, which we named HdcP, showing similarities to integral membrane transporters driving substrate/product exchange. The gene coding for HdcP was cloned downstream of a sequence specifying a histidine tag and expressed in Lactococcus lactis. The recombinant HdcP could drive the uptake of histidine into the cell and the exchange of histidine and histamine. The combination of HdcP and the histidine decarboxylase forms a typical bacterial decarboxylation pathway that may generate metabolic energy or be involved in the acid stress response. Analyses of sequences present in databases suggest that the other two proteins have dispensable functions. These results describe for the first time the genes encoding a histamine-producing pathway and provide clues to the parsimonious distribution and the instability of histamine-producing lactic acid bacteria. PMID:15746344

  11. Improving green roofs and rail road greening systems using Bacillus subtilis and Lactobacillus ssp.

    PubMed

    Grüneberg, H; Oschmann, C; Dunya, S; Ulrichs, C

    2006-01-01

    Aim of the present study was the improvement of existing methods for green roof and rail road greening systems using soil borne bacteria. Bacillus subtilis and Lactobacillus ssp. alone and in combination with vinasse applied to different growing substrates were tested. The substrates were brick chips, textile mats, mineral wool mats, and a commercial available substrate for the Swedish company VegTech. All four substrates were tested along an artificial rail track on the experimental station at Humboldt University Berlin, and partly on an existing rail track in Munich, Germany. Plants selected for the experiments belong to the genus Sedum, which is relatively tolerant to dry conditions. Inoculation of plants with bacteria had no effect on plant growth parameters and on coverage of different mobile bedding systems with Sedum plants. There was no significant difference between the various treatments in Munich. In both experiments, the addition of vinasse alone improved plant growth. Plant growth was significantly different on all substrates, whereas brick chips and the commercial roof soil was the best substrate. Brick chips are a cheap substrate which can be used for rail track greening. The results indicate that the quality of the substrate is the most important factor for remediation and greening of rail tracks and roof tops. The rapid growth of plants can be influenced by the application of vinasse as additional nutrient solution (potash (K) source) or nutrient enriched substrate. PMID:17390783

  12. Evaluation and functional characterization of a biosurfactant produced by Lactobacillus plantarum CFR 2194.

    PubMed

    Madhu, Arenahalli Ningegowda; Prapulla, Siddalingaiya Gurudutt

    2014-02-01

    The study details the investigations on the ability of Lactobacillus plantarum CFR 2194, an isolate from kanjika, a rice-based ayurvedic fermented product, to produce biosurfactant. Surfactant production, as a function of fermentation time, indicates that the maximum production occurred at 72 h under stationary conditions. Isolation, partial purification, and characterization of the biosurfactant produced have been carried out, and Fourier transform infrared spectroscopy (FTIR) spectra demonstrated that biosurfactants were constituted by protein and polysaccharide fractions, i.e., possessed the structure typical of glycoprotein, which is affected by the medium composition and the phase of growth of the biosurfactant-synthesizing strain. Critical micelle concentration (cmc) of the biosurfactant was found to be 6 g l(-1). The emulsification index (EI), emulsification activity (EA), and emulsion stability (ES) values of the biosurfactant have confirmed its emulsification property. Aqueous fractions of the produced biosurfactant exhibited a significant antimicrobial activity against the food-borne pathogenic species: Escherichia coli ATCC 31705, E. coli MTCC 108, Salmonella typhi, Yersinia enterocolitica MTCC 859, and Staphylococcus aureus F 722. More importantly, the biosurfactant from L. plantarum showed antiadhesive property against above food-borne pathogens. The results thus indicate the potential for developing strategies to prevent microbial colonization of food contact surfaces and health-care prosthesis using these biosurfactants. PMID:24258794

  13. Riboflavin-overproducing strains of Lactobacillus fermentum for riboflavin-enriched bread.

    PubMed

    Russo, Pasquale; Capozzi, Vittorio; Arena, Mattia Pia; Spadaccino, Giuseppina; Dueñas, María Teresa; López, Paloma; Fiocco, Daniela; Spano, Giuseppe

    2014-04-01

    Lactobacillus fermentum isolated from sourdough was able to produce riboflavin. Spontaneous roseoflavin-resistant mutants were obtained by exposing the wild strain (named L. fermentum PBCC11) to increasing concentrations of roseoflavin. Fifteen spontaneous roseoflavin-resistant mutants were isolated, and the level of vitamin B? was quantified by HPLC. Seven mutant strains produced concentrations of vitamin B? higher than 1 mg L?¹. Interestingly, three mutants were unable to overproduce riboflavin even though they were able to withstand the selective pressure of roseoflavin. Alignment of the rib leader region of PBCC11 and its derivatives showed only point mutations at two neighboring locations of the RFN element. In particular, the highest riboflavin-producing isolates possess an A to G mutation at position 240, while the lowest riboflavin producer carries a T to A substitution at position 236. No mutations were detected in the derivative strains that did not have an overproducing phenotype. The best riboflavin overproducing strain, named L. fermentum PBCC11.5, and its parental strain were used to fortify bread. The effect of two different periods of fermentation on the riboflavin level was compared. Bread produced using the coinoculum yeast and L. fermentum PBCC11.5 led to an approximately twofold increase of final vitamin B? content. PMID:24413973

  14. Dissipation of pirimiphos-methyl during wheat fermentation by Lactobacillus plantarum.

    PubMed

    Dor?evi?, T M; Siler-Marinkovi?, S S; Durovi?-Pej?ev, R D; Dimitrijevi?-Brankovi?, S I; Gaji? Umiljendi?, J S

    2013-11-01

    In this study, the dissipation of pirimiphos-methyl during wheat fermentation by Lactobacillus plantarum was investigated. Sample preparation for GC/MS detection of pirimiphos-methyl residues from fermented wheat substrate was carried out by two steps: extraction with 25 mL of methanol : acetone = 1 : 1 solvent mix for 30 min, followed by clean-up procedure through a glass column with florisil coupled with elution by 25 mL of ethyl acetate : acetone = 4 : 1. To obtain the highest pesticide degradation level, the fermentation conditions were optimized according to response surface methodology. Our results showed that L. plantarum was able to reduce the level of pirimiphos-methyl in wheat. Although pirimiphos-methyl was partially labile during sterilization prior inoculation (~37-50%), and there was also spontaneous chemical degradation of pesticide (~6-11%), overall L. plantarum enhanced degradation from 15 to 34%, that is, to nearly 81%. Additionally, the effect of pirimiphos-methyl on the lactobacilli growth, and efficiency of fermentation, was studied where pirimiphos-methyl inhibit the growth of bacteria in concentrations higher than 5 mg kg(-1), while the presence of pirimiphos-methyl did not overall affect the lactic acid fermentation. PMID:23879636

  15. Two-peptide bacteriocin PlnEF causes cell membrane damage to Lactobacillus plantarum.

    PubMed

    Zhang, Xu; Wang, Yang; Liu, Lei; Wei, Yunlu; Shang, Nan; Zhang, Xiangmei; Li, Pinglan

    2016-02-01

    Biologically active, artificially synthesized two-peptide bacteriocin PlnEF was used to study its mode of action on sensitive bacteria Lactobacillus plantarum pl2. The data obtained showed that PlnEF induced membrane permeabilization, allowing for the efflux of electrolytes, which was evidenced by the increased extracellular conductivity, the dissipation of transmembrane electrical potential and pH gradient, and rapid intracellular ATP depletion after L. plantarum pl2 cells were treated with PlnEF for minutes. Laser confocal microscopy showed that PlnEF accumulated very quickly in L. plantarum pl2 cells and the accumulation of PlnEF caused damage to cell membrane. Scanning electron microscopy and transmission electron microscopy further showed that PlnEF induced morphological changes and structure disruption to L. plantarum pl2 cells, such as the formation of blebs, microspheres, membrane deformation and cell lysis. In summary, the data obtained show that PlnEF caused cell membrane damage to L. plantarum pl2 cells. Our study reveals the antimicrobial mechanism of two-peptide bacteriocin PlnEF against L. plantarum. PMID:26615918

  16. Unique secreted–surface protein complex of Lactobacillus rhamnosus, identified by phage display

    PubMed Central

    Gagic, Dragana; Wen, Wesley; Collett, Michael A; Rakonjac, Jasna

    2013-01-01

    Proteins are the most diverse structures on bacterial surfaces; hence, they are candidates for species- and strain-specific interactions of bacteria with the host, environment, and other microorganisms. Genomics has decoded thousands of bacterial surface and secreted proteins, yet the function of most cannot be predicted because of the enormous variability and a lack of experimental data that would allow deduction of function through homology. Here, we used phage display to identify a pair of interacting extracellular proteins in the probiotic bacterium Lactobacillus rhamnosus HN001. A secreted protein, SpcA, containing two bacterial immunoglobulin-like domains type 3 (Big-3) and a domain distantly related to plant pathogen response domain 1 (PR-1-like) was identified by screening of an L. rhamnosus HN001 library using HN001 cells as bait. The SpcA-“docking” protein, SpcB, was in turn detected by another phage display library screening, using purified SpcA as bait. SpcB is a 3275-residue cell-surface protein that contains general features of large glycosylated Serine-rich adhesins/fibrils from gram-positive bacteria, including the hallmark signal sequence motif KxYKxGKxW. Both proteins are encoded by genes within a L. rhamnosus-unique gene cluster that distinguishes this species from other lactobacilli. To our knowledge, this is the first example of a secreted-docking protein pair identified in lactobacilli. PMID:23233310

  17. Change in cell surface properties of Lactobacillus casei under heat shock treatment.

    PubMed

    Haddaji, Najla; Mahdhi, Abdel Karim; Krifi, Boubaker; Ismail, Manel Ben; Bakhrouf, Amina

    2015-05-01

    We undertake this study in the aim to give new insight about the change in cellular physiological state under heat shock treatment and probiotic strain screening procedure. Different cell properties have been studied like adhesive ability to biotic and abiotic surfaces, the cell surface hydrophobicity and the fatty acids profiles. Compared to the normal cells, the heated cells increased their adhesive ability to biotic surface. However, the adhesion to abiotic surface was decreased. The cell surface hydrophobicity of the heated strains showed a significant decrease (P < 0.05). Our data revealed that high temperature change the fatty acids profiles of the treated cells, especially the proportions of unsaturated and saturated fatty acid. In fact, the ratio of saturated to unsaturated fatty acids of the heated Lactobacillus casei cells was significantly higher than that of the control cells (P < 0.05). The present finding could firstly add new insight about the response of probiotic to stressful conditions, such us the important role of cell membrane, considered as the first main structure to be damaged by physicochemical stress, in stress resistance because of their composition that can change in adaptation to harsh conditions. Secondly, there is no relationship between changes in membrane composition and fluidity induced by heat shock treatment and adhesion to biotic and abiotic surface. PMID:25825473

  18. Novel Method for Enumeration of Viable Lactobacillus plantarum WCFS1 Cells after Single-Droplet Drying

    PubMed Central

    Perdana, Jimmy; Bereschenko, Ludmila; Roghair, Mark; Fox, Martijn B.; Boom, Remko M.; Kleerebezem, Michiel

    2012-01-01

    Survival of probiotic bacteria during drying is not trivial. Survival percentages are very specific for each probiotic strain and can be improved by careful selection of drying conditions and proper drying carrier formulation. An experimental approach is presented, comprising a single-droplet drying method and a subsequent novel screening methodology, to assess the microbial viability within single particles. The drying method involves the drying of a single droplet deposited on a flat, hydrophobic surface under well-defined drying conditions and carrier formulations. Semidried or dried particles were subjected to rehydration, fluorescence staining, and live/dead enumeration using fluorescence microscopy. The novel screening methodology provided accurate survival percentages in line with conventional plating enumeration and was evaluated in single-droplet drying experiments with Lactobacillus plantarum WCFS1 as a model probiotic strain. Parameters such as bulk air temperatures and the carrier matrices (glucose, trehalose, and maltodextrin DE 6) were varied. Following the experimental approach, the influence on the viability as a function of the drying history could be monitored. Finally, the applicability of the novel viability assessment was demonstrated for samples obtained from drying experiments at a larger scale. PMID:22983965

  19. Development of a Potential Probiotic Fresh Cheese Using Two Lactobacillus salivarius Strains Isolated from Human Milk

    PubMed Central

    Cárdenas, Nivia; Peirotén, Ángela; Rodríguez, Juan M.; Fernández, Leónides

    2014-01-01

    Cheeses have been proposed as a good alternative to other fermented milk products for the delivery of probiotic bacteria to the consumer. The objective of this study was to assess the survival of two Lactobacillus salivarius strains (CECT5713 and PS2) isolated from human milk during production and storage of fresh cheese for 28 days at 4°C. The effect of such strains on the volatile compounds profile, texture, and other sensorial properties, including an overall consumer acceptance, was also investigated. Both L. salivarius strains remained viable in the cheeses throughout the storage period and a significant reduction in their viable counts was only observed after 21 days. Globally, the addition of the L. salivarius strains did not change significantly neither the chemical composition of the cheese nor texture parameters after the storage period, although cheeses manufactured with L. salivarius CECT5713 presented significantly higher values of hardness. A total of 59 volatile compounds were identified in the headspace of experimental cheeses, and some L. salivarius-associated differences could be identified. All cheeses presented good results of acceptance after the sensory evaluation. Consequently, our results indicated that fresh cheese can be a good vehicle for the two L. salivarius strains analyzed in this study. PMID:24971351

  20. Association of Lactobacillus crispatus with fructo-oligosaccharides and ascorbic acid in hydroxypropyl methylcellulose vaginal insert.

    PubMed

    Vitali, Beatrice; Abruzzo, Angela; Parolin, Carola; Palomino, Rogers Alberto Ñahui; Dalena, Francesco; Bigucci, Federica; Cerchiara, Teresa; Luppi, Barbara

    2016-01-20

    The aim of this work was to develop a synbiotic vaginal insert containing the probiotic strain Lactobacillus crispatus BC5, the prebiotic substrate fructo-oligosaccharide and the antioxidant agent ascorbic acid, for the prophylaxis and therapy of vaginal infections. Mucoadhesive in situ gelling vaginal inserts based on hydroxypropyl methylcellulose were prepared by freeze-drying, stored at +2-8°C for 90 days and characterized in terms of technological and functional properties. Complete survival of L. crispatus BC5 was found immediately after insert preparation (96.08%) as well as after 90 days of storage (95.82%) in the vaginal inserts containing fructo-oligosaccharide, ascorbic acid and skimmed milk. Synbiotic inserts showed improved mucoadhesion ability (from three- to five-fold) with respect to a standard formulation based on hydroxypropyl methylcellulose alone. Moreover, inserts allowed to modulate lactobacilli release in virtue of the different amounts of fructo-oligosaccharide. Finally, antimicrobial activity was exerted by L. crispatus BC5 released from the vaginal formulation. PMID:26572459

  1. Microencapsulation of Bifidobacterium animalis subsp. lactis and Lactobacillus acidophilus in cocoa butter using spray chilling technology

    PubMed Central

    Pedroso, D.L.; Dogenski, M.; Thomazini, M.; Heinemann, R.J.B.; Favaro-Trindade, C.S.

    2013-01-01

    In the present study, the cells of Bifidobacterium animalis subsp. lactis (BI-01) and Lactobacillus acidophilus (LAC-04) were encapsulated in cocoa butter using spray-chilling technology. Survival assays were conducted to evaluate the resistance of the probiotics to the spray-chilling process, their resistance to the simulated gastric and intestinal fluids (SGF and SIF), and their stability during 90 days of storage. The viability of the cells was not affected by microencapsulation. The free and encapsulated cells of B. animalis subsp. lactis were resistant to both SGF and SIF. The micro-encapsulated cells of L. acidophilus were more resistant to SGF and SIF than the free cells; the viability of the encapsulated cells was enhanced by 67%, while the free cells reached the detection limit of the method (103 CFU/g). The encapsulated probiotics were unstable when they were stored at 20 °C. The population of encapsulated L. acidophilus decreased drastically when they were stored at 7 °C; only 20% of cells were viable after 90 days of storage. The percentage of viable cells of the encapsulated B. animalis subsp.lactis, however, was 72% after the same period of storage. Promising results were obtained when the microparticles were stored at ?18 °C; the freeze granted 90 days of shelf life to the encapsulated cells. These results suggest that the spray-chilling process using cocoa butter as carrier protects L. acidophilus from gastrointestinal fluids. However, the viability of the cells during storage must be improved. PMID:24516445

  2. Lactobacillus rhamnosus GG Lysate Increases Re-Epithelialization of Keratinocyte Scratch Assays by Promoting Migration

    PubMed Central

    Mohammedsaeed, Walaa; Cruickshank, Sheena; McBain, Andrew J.; O’Neill, Catherine A.

    2015-01-01

    A limited number of studies have investigated the potential of probiotics to promote wound healing in the digestive tract. The aim of the current investigation was to determine whether probiotic bacteria or their extracts could be beneficial in cutaneous wound healing. A keratinocyte monolayer scratch assay was used to assess re-epithelialization; which comprises keratinocyte proliferation and migration. Primary human keratinocyte monolayers were scratched then exposed to lysates of Lactobacillus (L) rhamnosus GG, L. reuteri, L. plantarum or L. fermentum. Re-epithelialization of treated monolayers was compared to that of untreated controls. Lysates of L. rhamnosus GG and L. reuteri significantly increased the rate of re-epithelialization, with L. rhamnosus GG being the most efficacious. L. reuteri increased keratinocyte proliferation while L. rhamnosus GG lysate significantly increased proliferation and migration. Microarray analysis of L. rhamnosus GG treated scratches showed increased expression of multiple genes including the chemokine CXCL2 and its receptor CXCR2. These are involved in normal wound healing where they stimulate keratinocyte proliferation and/or migration. Increased protein expression of both CXCL2 and CXCR2 were confirmed by ELISA and immunoblotting. These data demonstrate that L. rhamnosus GG lysate accelerates re-epithelialization of keratinocyte scratch assays, potentially via chemokine receptor pairs that induce keratinocyte migration. PMID:26537246

  3. Preparation and characterization of a coacervate extended-release microparticulate delivery system for Lactobacillus rhamnosus

    PubMed Central

    Alli, Sk Md Athar

    2011-01-01

    Background: The purpose of this study was to develop a mucoadhesive coacervate microparticulate system to deliver viable Lactobacillus rhamnosus cells into the gut for an extended period of time while maintaining high numbers of viable cells within the formulation throughout its shelf-life and during gastrointestinal transit. Methods: Core coacervate mucoadhesive microparticles of L. rhamnosus were developed using several grades of hypromellose and were subsequently enteric-coated with hypromellose phthalate. Microparticles were evaluated for percent yield, entrapment efficiency, surface morphology, particle size, size distribution, zeta potential, flow properties, in vitro swelling, mucoadhesion properties, in vitro release profile and release kinetics, in vivo probiotic activity, and stability. The values for the kinetic constant and release exponent of model-dependent approaches, the difference factor, similarity factor, and Rescigno indices of model-independent approaches were determined for analyzing in vitro dissolution profiles. Results: Experimental microparticles of formulation batches were of spherical shape with percent yields of 41.24%–58.18%, entrapment efficiency 45.18%–64.16%, mean particle size 33.10–49.62 ?m, and zeta potential around ?11.5 mV, confirming adequate stability of L. rhamnosus at room temperature. The in vitro L. rhamnosus release profile follows zero-order kinetics and depends on the grade of hypromellose and the L. rhamnosus to hypromellose ratio. Conclusion: Microparticles delivered L. rhamnosus in simulated intestinal conditions for an extended period, following zero-order kinetics, and exhibited appreciable mucoadhesion in simulated intestinal conditions. PMID:21984867

  4. Optimization and partial characterization of bacteriocin produced by Lactobacillus bulgaricus -TLBFT06 isolated from Dahi.

    PubMed

    Mahmood, Talat; Masud, Tariq; Ali, Sartaj; Abbasi, Kashif Sarfraz; Liaquat, Muhammad

    2015-03-01

    Lactobacillus bulgaricus is one of the predominant lactic acid bacteria of dahi, conferring technological and functional attributes. In the present study thirty dahi samples were investigated for bacteriocin producing L. bulgaricus. Fourteen different isolates were obtained and five were scrutinized for antibacterial activities against food born pathogens. Amongst, a strain TLB06FT was found to have a wide array of antibacterial activities against Gram positive and negative bacteria was selected for further characterization. Growth media optimization for this strain revealed maximum bacteriocin production on MRS media supplemented with glucose (2%), sodium chloride (1%), Tween-80 (0.5%) and yeast extract (1 %). In addition, optimization of growth conditions revealed maximum bacteriocin production at pH 5.5 and temperature of 30-37°C. Bacteriocin showed thermo stability at 90°C and remained highly active in the pH range of 3.5-7.5, inactive by protein catalyzing enzymes and showed no change in activity (800AumL(-1)) when treated with organic solvents and surfactants. The obtained bacteriocin was purified to 1600AU mL(-1) by ammonium sulfate precipitation (80%) by using dialyzing tubing. In the same way, a single peak was obtained by RP-HPLC having antibacterial activity of 6400AU mL(-1). Thus, wild strains of L. bulgaricus have great potential for the production new and novel type of bacteriocins. PMID:25730789

  5. Antimicrobial activity of reuterin produced by Lactobacillus reuteri on Listeria monocytogenes in cold-smoked salmon.

    PubMed

    Montiel, R; Martín-Cabrejas, I; Langa, S; El Aouad, N; Arqués, J L; Reyes, F; Medina, M

    2014-12-01

    Lactobacillus reuteri INIA P579 was used for the production and purification of reuterin. The purity of reuterin was assessed by high resolution electrospray ionization mass spectrometry (HRESIMS) and nuclear magnetic resonance (NMR) spectroscopy. After purification, reuterin concentration obtained was 1.3 M. The inhibitory activity using Escherichia coli K12 as indicator strain was estimated to be 510 AU/ml. Survival curves in tryptic soy broth revealed that reuterin required to inhibit the growth of three Listeria monocytogenes strains was in the range of 2-4 AU/ml. Purified reuterin (10 AU/g) significantly reduced the growth of L. monocytogenes in cold-smoked salmon kept under moderate or strong temperature abuse conditions. After 15 d at 8 °C, cold-smoked salmon with added reuterin exhibited L. monocytogenes counts 2.0 log CFU/g lower than control smoked salmon with no reuterin added. At 30 °C, reuterin also controlled the growth of the pathogen, with counts 1.4 and 0.9 log CFU/g lower than those observed in control smoked salmon after 24 and 48 h, respectively. The addition of purified reuterin might be used as a hurdle technology to improve the safety and extend the shelf-life of lightly preserved seafood products such as cold-smoked salmon. PMID:25084638

  6. Lactobacillus paracasei CBA L74 interferes with gliadin peptides entrance in Caco-2 cells.

    PubMed

    Sarno, Marco; Lania, Giuliana; Cuomo, Marialaura; Nigro, Federica; Passannanti, Francesca; Budelli, Andrea; Fasano, Francesca; Troncone, Riccardo; Auricchio, Salvatore; Barone, Maria Vittoria; Nigro, Roberto; Nanayakkara, Merlin

    2014-12-01

    Several recent reports describe a role of probiotics as a therapeutic approach for celiac disease (CD). Two undigested A-gliadin peptides, P31-43 and P57-68, are central to CD pathogenesis, inducing an innate and an adaptive immune response, respectively. They enter enterocytes and localize to vesicular compartment to induce their toxic/immunogenics effects. In this article, we tested the effect of probiotic Lactobacillus paracasei (LP) CBA L74 (International Depository Accession Number LMG P-24778), its supernatant and LP-fermented cereals on gliadin peptides, P31-43 and P57-68, entrance in Caco-2 cells. Both LP CBA L74 and its supernatant inhibit P31-43 (intensity of fluorescence; FI: 75%) and P57-68 (FI: 50%) entrance in Caco2 cells, indicating that this biological effect is due to some product included in LP CBA L74 supernatant. This effect was present also after fermentation of cereals. This study describes a novel effect of probiotics in the prevention of undigested gliadin peptides toxic effects. PMID:25030417

  7. Protective Effects of Lactobacillus plantarum CCFM8246 against Copper Toxicity in Mice.

    PubMed

    Tian, Fengwei; Xiao, Yue; Li, Xiaoxiao; Zhai, Qixiao; Wang, Gang; Zhang, Qiuxiang; Zhang, Hao; Chen, Wei

    2015-01-01

    Lactobacillus plantarum CCFM8246, which has a relatively strong copper binding capacity and tolerance to copper ions, was obtained by screening from 16 lactic acid bacteria in vitro. The selected strain was then applied to a mouse model to evaluate its protective function against copper intoxication in vivo. The experimental mice were divided into an intervention group and a therapy group; mice in the intervention group received co-administration of CCFM8246 and a copper ion solution by gavage, while mice in the therapy group were treated with CCFM8246 after 4 weeks of copper exposure. In both two groups, mice treated with copper alone and that treated with neither CCFM8246 nor copper served as positive and negative controls, respectively. At the end of the experimental period, the copper content in feces and tissues, the activity of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum, and oxidation stress indices in liver and kidney tissue were determined. Learning and memory ability was evaluated by Morris water maze experiments. The results indicated that treatment with CCFM8246 significantly increased the copper content in feces to promote copper excretion, reduce the accumulation of copper in tissues, reverse oxidative stress induced by copper exposure, recover the ALT and AST in serum and improve the spatial memory of mice. PMID:26605944

  8. Lactobacillus plantarum vaccine vector expressing hemagglutinin provides protection against H9N2 challenge infection.

    PubMed

    Shi, Shao-Hua; Yang, Wen-Tao; Yang, Gui-Lian; Zhang, Xu-Ke; Liu, Yu-Ying; Zhang, Li-Jiao; Ye, Li-Ping; Hu, Jing-Tao; Xin Xing; Qi, Chong; Li, Yu; Wang, Chun-Feng

    2016-01-01

    Hemagglutinin (HA) has been demonstrated as an effective candidate vaccine antigen against AIVs. Dendritic cell-targeting peptide (DCpep) can enhance the robustness of immune responses. The purpose of this study was to evaluate whether DCpep could enhance the immune response against H9N2 AIV when utilizing Lactobacillus plantarum NC8 (NC8) to present HA-DCpep in mouse and chicken models. To accomplish this, a mucosal vaccine of a recombinant NC8 strain expressing HA and DCpep that was constructed in a previous study was employed. Orally administered NC8-pSIP409-HA-DCpep elicited high serum titers of hemagglutination-inhibition (HI) antibodies in mice and also induced robust T cell immune responses in both mouse and chicken models. Orally administered NC8-pSIP409-HA-DCpep elicited high serum titers of hemagglutination-inhibition (HI) antibodies in mice and also induced robust T cell immune responses in both mouse and chicken models. These results revealed that recombinant L. plantarum NC8-pSIP409-HA-DCpep is an effective vaccine candidate against H9N2 AIVs. PMID:26363195

  9. Inactivation of Lactobacillus rhamnosus GG by fixation modifies its probiotic properties.

    PubMed

    Markowicz, C; Kubiak, P; Grajek, W; Schmidt, M T

    2016-01-01

    Probiotics are microorganisms that have beneficial effects on the host and are safe for oral intake in a suitable dose. However, there are situations in which the administration of living microorganisms poses a risk for immunocompromised host. The objective of this study was to evaluate the influence of several fixation methods on selected biological properties of Lactobacillus rhamnosus GG that are relevant to its probiotic action. Fixation of the bacterial cells with ethanol, 2-propanol, glutaraldehyde, paraformaldehyde, and heat treatment resulted in a significant decrease of alkaline phosphatase, peroxidase, and ?-galactosidase activities. Most of the fixation procedures reduced bacterial cell hydrophobicity and increased adhesion capacity. The fixation procedures resulted in a different perception of the bacterial cells by enterocytes, which was shown as changes in gene expression in enterocytes. The results show that some procedures of inactivation allow a fraction of the enzymatic activity to be maintained. The adhesion properties of the bacterial cells were enhanced, but the response of enterocytes to fixed cells was different than to live bacteria. Inactivation allows maintenance and modification of some of the properties of the bacterial cells. PMID:26634746

  10. Enhancing vitamin B12 content in soy-yogurt by Lactobacillus reuteri.

    PubMed

    Gu, Qing; Zhang, Chen; Song, Dafeng; Li, Ping; Zhu, Xuan

    2015-08-01

    More attention from the aged and vegetarians has been paid to soy-product due to its taste, easy digestibility, as well as the association with health. However, soy-product has a defect of low vitamin content, mainly the water-soluble vitamin B12. This study was to investigate co-fermentation of glycerol and fructose in soy-yogurt to enhance vitamin B12 production by Lactobacillus reuteri. After a serial combination experiments, the co-fermentation was confirmed to enhance the production of vitamin B12 up to 18 ?g/100mL. Both supplementations induced the expression of cobT and cbiA and functioned to balance the redox reaction. Meanwhile, high content of fructose supplementation reduced the production of vitamin B12 and suppressed expression of cobT in bacteria. It was proved that the vitamin B12 content of this soy-yogurt is higher than other fermented soybean based food and thus can be served as an alternative food for the aged and vegetarians. PMID:25955289

  11. Positive selection on D-lactate dehydrogenases of Lactobacillus delbrueckii subspecies bulgaricus.

    PubMed

    Zhang, Jifeng; Gong, Guangyu; Wang, Xiao; Zhang, Hao; Tian, Weidong

    2015-08-01

    Lactobacillus delbrueckii has been widely used for yogurt fermentation. It has genes encoding both D- and L-type lactate dehydrogenases (LDHs) that catalyse the production of L(+) or D(-) stereoisomer of lactic acid. D-lactic acid is the primary lactate product by L. delbrueckii, yet it cannot be metabolised by human intestine. Since it has been domesticated for long time, an interesting question arises regarding to whether the selection pressure has affected the evolution of both L-LDH and D-LDH genes in the genome. To answer this question, in this study the authors first investigated the evolution of these two genes by constructing phylogenetic trees. They found that D-LDH-based phylogenetic tree could better represent the phylogenetic relationship in the acidophilus complex than L-LDH-based tree. They next investigated the evolutions of LDH genes of L. delbrueckii at amino acid level, and found that D-LDH gene in L. delbrueckii is positively selected, possibly a consequence of long-term domestication. They further identified four amino acids that are under positive selection. One of them, V261, is located at the centre of three catalytic active sites, indicating likely functional effects on the enzyme activity. The selection from the domestication process thus provides direction for future engineering of D-LDH. PMID:26243834

  12. The sim Operon Facilitates the Transport and Metabolism of Sucrose Isomers in Lactobacillus casei ATCC 334?

    PubMed Central

    Thompson, John; Jakubovics, Nicholas; Abraham, Bindu; Hess, Sonja; Pikis, Andreas

    2008-01-01

    Inspection of the genome sequence of Lactobacillus casei ATCC 334 revealed two operons that might dissimilate the five isomers of sucrose. To test this hypothesis, cells of L. casei ATCC 334 were grown in a defined medium supplemented with various sugars, including each of the five isomeric disaccharides. Extracts prepared from cells grown on the sucrose isomers contained high levels of two polypeptides with Mrs of ?50,000 and ?17,500. Neither protein was present in cells grown on glucose, maltose or sucrose. Proteomic, enzymatic, and Western blot analyses identified the ?50-kDa protein as an NAD+- and metal ion-dependent phospho-?-glucosidase. The oligomeric enzyme was purified, and a catalytic mechanism is proposed. The smaller polypeptide represented an EIIA component of the phosphoenolpyruvate-dependent sugar phosphotransferase system. Phospho-?-glucosidase and EIIA are encoded by genes at the LSEI_0369 (simA) and LSEI_0374 (simF) loci, respectively, in a block of seven genes comprising the sucrose isomer metabolism (sim) operon. Northern blot analyses provided evidence that three mRNA transcripts were up-regulated during logarithmic growth of L. casei ATCC 334 on sucrose isomers. Internal simA and simF gene probes hybridized to ?1.5- and ?1.3-kb transcripts, respectively. A 6.8-kb mRNA transcript was detected by both probes, which was indicative of cotranscription of the entire sim operon. PMID:18310337

  13. Effect of Low Shear Modeled Microgravity (LSMMG) on the Probiotic Lactobacillus Acidophilus ATCC 4356

    NASA Technical Reports Server (NTRS)

    Stahl, S.; Voorhies, A.; Lorenzi, H.; Castro-Wallace, S.; Douglas, G.

    2016-01-01

    The introduction of generally recognized as safe (GRAS) probiotic microbes into the spaceflight food system has the potential for use as a safe, non-invasive, daily countermeasure to crew microbiome and immune dysregulation. However, the microgravity effects on the stress tolerances and genetic expression of probiotic bacteria must be determined to confirm translation of strain benefits and to identify potential for optimization of growth, survival, and strain selection for spaceflight. The work presented here demonstrates the translation of characteristics of a GRAS probiotic bacteria to a microgravity analog environment. Lactobacillus acidophilus ATCC 4356 was grown in the low shear modeled microgravity (LSMMG) orientation and the control orientation in the rotating wall vessel (RWV) to determine the effect of LSMMG on the growth, survival through stress challenge, and gene expression of the strain. No differences were observed between the LSMMG and control grown L. acidophilus, suggesting that the strain will behave similarly in spaceflight and may be expected to confer Earth-based benefits.

  14. Oral administration of Lactobacillus plantarum HY7714 protects hairless mouse against ultraviolet B-induced photoaging.

    PubMed

    Kim, Hyun Mee; Lee, Dong Eun; Park, Soo Dong; Kim, Yong-Tae; Kim, Yu Jin; Jeong, Ji Woong; Jang, Sung Sik; Ahn, Young-Tae; Sim, Jae-Hun; Huh, Chul-Sung; Chung, Dae Kyun; Lee, Jung-Hee

    2014-11-28

    Ultraviolet (UV) irradiation alters multiple molecular pathways in the skin, thereby inducing skin damage, including photoaging. In recent years, probiotics have gained interest due to their beneficial effects on skin health, such as inhibiting atopic dermatitis and improving skin immunity or inflammation. However, little is known about the effects of probiotics on UVBinduced photoaging. In this study, we evaluated the effect of Lactobacillus plantarum HY7714 against UVB-induced photoaging in human dermal fibroblasts and hairless mice. The results showed that L. plantarum HY7714 treatment effectively rescued UVB-reduced procollagen expression through the inhibition of UVB-induced matrix metalloproteinase (MMP)-1 expression in human dermal fibroblasts. Data from a western blot showed that L. plantarum HY7714 inhibited the phosphorylation of Jun N-terminal kinase, thereby suppressing the UVB-induced phosphorylation and expression of c-Jun. Oral administration of L. plantarum HY7714 clearly inhibited the number, depth, and area of wrinkles in hairless mouse skin. Histological data showed that L. plantarum HY7714 significantly inhibited UVB-induced epidermal thickness in mice. Western blot and zymography data also revealed that L. plantarum HY7714 effectively inhibited MMP-13 expression as well as MMP-2 and -9 activities in dermal tissue. Collectively, these results provide further insight regarding the skin biological actions of L. plantarum HY7714, a potential skin anti-photoaging agent. PMID:25112318

  15. Protective Effects of Lactobacillus plantarum CCFM8610 against Acute Cadmium Toxicity in Mice

    PubMed Central

    Zhai, Qixiao; Wang, Gang; Zhao, Jianxin; Liu, Xiaoming; Zhang, Hao

    2013-01-01

    This study evaluated the protective effects of Lactobacillus plantarum CCFM8610, a selected probiotic with good cadmium binding capacity, against acute cadmium toxicity in mice. Ninety mice were divided into prevention and therapy groups. In the prevention groups, CCFM8610 was administered at 109 CFU once daily for 7 days, followed by a single oral dose of cadmium chloride at 1.8 mg cadmium for each mouse. In the therapy groups, the same dose of CCFM8610 was administered for 2 days after an identical single dose of cadmium exposure. Mice that received neither cadmium nor culture or that received cadmium alone served as negative and positive controls, respectively. The effects of both living and dead CCFM8610 on cadmium ion concentrations in feces, liver, and kidney were determined. Moreover, the alterations in reduced glutathione (GSH), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and histopathology in the liver and kidney were investigated. The results showed that compared to the mice that received cadmium only, CCFM8610 treatment can effectively decrease intestinal cadmium absorption, reduce tissue cadmium accumulation, alleviate renal and hepatic oxidative stress, and ameliorate hepatic histopathological changes. Living CCFM8610 administered after cadmium exposure offered the most significant protection. Our results suggested that CCFM8610 is more effective against acute cadmium toxicity than a simple antioxidant treatment due to its special physiological functions and that it can be considered a new dietary therapeutic strategy against acute cadmium toxicity. PMID:23263961

  16. Isolation and Characterization of a Novel Virulent Phage of Lactobacillus casei ATCC 393.

    PubMed

    Zhang, Xi; Lan, Yu; Jiao, Wenchao; Li, Yijing; Tang, Lijie; Jiang, Yanping; Cui, Wen; Qiao, Xinyuan

    2015-12-01

    A new virulent phage (Lcb) of Lactobacillus casei ATCC 393 was isolated from Chinese sauerkraut. It was specific to L. casei ATCC 393. Electron micrograph revealed that it had an icosahedral head (60.2 ± 0.8 nm in diameter) and a long tail (251 ± 2.6 nm). It belonged to the Siphoviridae family. The genome of phage Lcb was estimated to be approximately 40 kb and did not contain cohesive ends. One-step growth kinetics of its lytic development revealed latent and burst periods of 75 and 45 min, respectively, with a burst size of 16 PFU per infected cell. The phage was able to survive in a pH range between 4 and 11. However, a treatment of 70 °C for 30 min and 75 % ethanol or isopropanol for 20 min was observed to inactivate phage Lcb thoroughly. The presence of both Ca(2+) and Mg(2+) showed a little influence on phage adsorption, but they were indispensable to gain complete lysis and improve plaque formation. The adsorption kinetics were similar on viable or nonviable cells, and high adsorption rates maintained between 10 and 37 °C. The highest adsorption rate was at 30 °C. This study increased the knowledge on phages of L. casei. The characterization of phage Lcb is helpful to establish a basis for adopting effective strategies to control phage attack in industry. PMID:26123178

  17. Immunobiotic Lactobacillus rhamnosus improves resistance of infant mice against respiratory syncytial virus infection.

    PubMed

    Chiba, Eriko; Tomosada, Yohsuke; Vizoso-Pinto, Maria Guadalupe; Salva, Susana; Takahashi, Takuya; Tsukida, Kohichiro; Kitazawa, Haruki; Alvarez, Susana; Villena, Julio

    2013-10-01

    Previously we showed that orally administered Lactobacillus rhamnosus CRL1505 beneficially regulated the balance between pro- and anti-inflammatory mediators in the lungs of poly(I:C)-challenged mice, allowing an effective inflammatory response against the TLR3/RIG-I agonist but at the same time reducing tissue damage. The aim of the present study was to investigate whether oral administration of the CRL1505 strain was able to improve resistance against respiratory syncytial virus (RSV) infection in infant mice and to evaluate the immunological mechanisms involved in the immunobiotic effect. We demonstrated that treatment of 3-week old BALB/c mice with L. rhamnosus CRL1505 significantly reduce lung viral loads and tissue injuries after the challenge with RSV. Moreover, we showed that the protective effect achieved by the CRL1505 strain is related to its capacity to differentially modulate respiratory antiviral immune response. Our results shows that IFN-? and IL-10 secreted in response to L. rhamnosus CRL1505 oral stimulation would modulate the pulmonary innate immune microenvironment conducting to the activation of CD103(+) and CD11b(high) dendritic cells and the generation of CD3(+)CD4(+)IFN-?(+) Th1 cells with the consequent attenuation of the strong and damaging Th2 reactions associated with RSV challenge. Our results indicate that modulation of the common mucosal immune system by immunobiotics could favor protective immunity against respiratory viral pathogens with a high attack rate in early infancy, such as RSV. PMID:23838113

  18. Radio-protective effect of polysaccharides isolated from Lactobacillus brevis-fermented Ecklonia cava.

    PubMed

    Lee, WonWoo; Ahn, Ginnae; Lee, Bae-Jin; Wijesinghe, W A J P; Kim, Daekyung; Yang, Hyemi; Kim, Young Mog; Park, Sun Joo; Jee, Youngheun; Jeon, You-Jin

    2013-01-01

    We investigated the radioprotective effects of a polysaccharide isolated from enzymatic extracts of Ecklonia cava (E. cava) fermented by fungi and bacteria. We identified that the aqueous extract of the Lactobacillus brevis-fermented E. cava especially showed the highest proliferation effect. In addition, the enzymatic extract prepared by enzyme-assisted extraction using Viscozyme (VLFE) significantly increased cell proliferation. Further study indicated that the polysaccharides isolated from the >30 kDa fraction of VLFE (VLFEP) significantly enhanced survival and proliferation effects in ?-ray-irradiated cells. Also, VLFEP markedly reduced the DNA damage, production of reactive oxygen species, and the percentage of Sub-G(1) DNA contents caused by ?-ray-irradiation. Moreover, VLFEP modulated the expression levels of p53, Bax, and Bcl-2 via inhibition of I?B? degradation and phosphorylation and NF?B p65 translocation into nuclei. These results demonstrate that VLFEP has radioprotective properties including the modulation of apoptosis via the inhibition of the NF?B signaling pathway. PMID:23068138

  19. Enhanced expression of codon optimized Mycobacterium avium subsp. paratuberculosis antigens in Lactobacillus salivarius

    PubMed Central

    Johnston, Christopher D.; Bannantine, John P.; Govender, Rodney; Endersen, Lorraine; Pletzer, Daniel; Weingart, Helge; Coffey, Aidan; O'Mahony, Jim; Sleator, Roy D.

    2014-01-01

    It is well documented that open reading frames containing high GC content show poor expression in A+T rich hosts. Specifically, G+C-rich codon usage is a limiting factor in heterologous expression of Mycobacterium avium subsp. paratuberculosis (MAP) proteins using Lactobacillus salivarius. However, re-engineering opening reading frames through synonymous substitutions can offset codon bias and greatly enhance MAP protein production in this host. In this report, we demonstrate that codon-usage manipulation of MAP2121c can enhance the heterologous expression of the major membrane protein (MMP), analogous to the form in which it is produced natively by MAP bacilli. When heterologously over-expressed, antigenic determinants were preserved in synthetic MMP proteins as shown by monoclonal antibody mediated ELISA. Moreover, MMP is a membrane protein in MAP, which is also targeted to the cellular surface of recombinant L. salivarius at levels comparable to MAP. Additionally, we previously engineered MAP3733c (encoding MptD) and show herein that MptD displays the tendency to associate with the cytoplasmic membrane boundary under confocal microscopy and the intracellularly accumulated protein selectively adheres to the MptD-specific bacteriophage fMptD. This work demonstrates there is potential for L. salivarius as a viable antigen delivery vehicle for MAP, which may provide an effective mucosal vaccine against Johne's disease. PMID:25237653

  20. Lactobacillus rhamnosus GG-supplemented formula expands butyrate-producing bacterial strains in food allergic infants

    DOE PAGESBeta

    Berni Canani, Roberto; Sangwan, Naseer; Stefka, Andrew T.; Nocerino, Rita; Paparo, Lorella; Aitoro, Rosita; Calignano, Antonio; Khan, Aly A.; Gilbert, Jack A.; Nagler, Cathryn R.

    2015-09-22

    Dietary intervention with extensively hydrolyzed casein formula supplemented with Lactobacillus rhamnosus GG (EHCF+LGG) accelerates tolerance acquisition in infants with cow’s milk allergy (CMA). We examined whether this effect is attributable, at least in part, to an influence on the gut microbiota. Fecal samples from healthy controls (n=20) and from CMA infants (n=19) before and after treatment with EHCF with (n=12) and without (n=7) supplementation with LGG were compared by 16S rRNA-based operational taxonomic unit clustering and oligotyping. Differential feature selection and generalized linear model fitting revealed that the CMA infants have a diverse gut microbial community structure dominated by Lachnospiraceaemore »(20.5±9.7%) and Ruminococcaceae (16.2±9.1%). Blautia, Roseburia and Coprococcus were significantly enriched following treatment with EHCF and LGG, but only one genus, Oscillospira, was significantly different between infants that became tolerant and those that remained allergic. However, most tolerant infants showed a significant increase in fecal butyrate levels, and those taxa that were significantly enriched in these samples, Blautia and Roseburia, exhibited specific strain-level demarcations between tolerant and allergic infants. As a result, our data suggest that EHCF+LGG promotes tolerance in infants with CMA, in part, by influencing the strain-level bacterial community structure of the infant gut.« less

  1. Fermentation of quinoa and wheat slurries by Lactobacillus plantarum CRL 778: proteolytic activity.

    PubMed

    Dallagnol, Andrea Micaela; Pescuma, Micaela; De Valdez, Graciela Font; Rollán, Graciela

    2013-04-01

    Quinoa fermentation by lactic acid bacteria (LAB) is an interesting alternative to produce new bakery products with high nutritional value; furthermore, they are suitable for celiac patients because this pseudo-cereal contains no gluten. Growth and lactic acid production during slurry fermentations by Lactobacillus plantarum CRL 778 were greater in quinoa (9.8 log?cfu/mL, 23.1 g/L) than in wheat (8.9 log?cfu/mL, 13.9 g/L). Lactic fermentation indirectly stimulated flour protein hydrolysis by endogenous proteases of both slurries. However, quinoa protein hydrolysis was faster, reaching 40-100% at 8 h of incubation, while wheat protein hydrolysis was only 0-20%. In addition, higher amounts of peptides (24) and free amino acids (5 g/L) were determined in quinoa compared to wheat. Consequently, greater concentrations (approx. 2.6-fold) of the antifungal compounds (phenyllactic and hydroxyphenyllactic acids) were synthesized from Phe and Tyr in quinoa by L. plantarum CRL 778, an antifungal strain. These promising results suggest that this LAB strain could be used in the formulation of quinoa sourdough to obtain baked goods with improved nutritional quality and shelf life, suitable for celiac patients. PMID:23129182

  2. Characterisation of biofilms formed by Lactobacillus plantarum WCFS1 and food spoilage isolates.

    PubMed

    Fernández Ramírez, Mónica D; Smid, Eddy J; Abee, Tjakko; Nierop Groot, Masja N

    2015-08-17

    Lactobacillus plantarum has been associated with food spoilage in a wide range of products and the biofilm growth mode has been implicated as a possible source of contamination. In this study we analysed the biofilm forming capacity of L. plantarum WCFS1 and six food spoilage isolates. Biofilm formation as quantified by crystal violet staining and colony forming units was largely affected by the medium composition, growth temperature and maturation time and by strain specific features. All strains showed highest biofilm formation in Brain Heart Infusion medium supplemented with manganese and glucose. For L. plantarum biofilms the crystal violet (CV) assay, that is routinely used to quantify total biofilm formation, correlates poorly with the number of culturable cells in the biofilm. This can in part be explained by cell death and lysis resulting in CV stainable material, conceivably extracellular DNA (eDNA), contributing to the extracellular matrix. The strain to strain variation may in part be explained by differences in levels of eDNA, likely as result of differences in lysis behaviour. In line with this, biofilms of all strains tested, except for one spoilage isolate, were sensitive to DNase treatment. In addition, biofilms were highly sensitive to treatment with Proteinase K suggesting a role for proteins and/or proteinaceous material in surface colonisation. This study shows the impact of a range of environmental factors and enzyme treatments on biofilm formation capacity for selected L. plantarum isolates associated with food spoilage, and may provide clues for disinfection strategies in food industry. PMID:25965141

  3. Suitability of Bifidobacterium spp. and Lactobacillus plantarum as probiotics intended for fruit juices containing citrus extracts.

    PubMed

    Bevilacqua, Antonio; Campaniello, Daniela; Corbo, Maria Rosaria; Maddalena, Lucia; Sinigaglia, Milena

    2013-11-01

    A strain of Lactobacillus plantarum and 4 strains of bifidobacteria were inoculated in apple juice and in a commercial beverage labeled as "red-fruit juice," containing citrus extracts as natural preservatives; the suitability of the probiotics was evaluated in relation to their resistance to 2 kinds of citrus extracts (biocitro and lemon extract), survival in juices at 4 and 37 °C, and inhibition of Zygosaccharomyces bailii. Cell count of L. plantarum and bifidobacteria over time was fitted through the Weibull equation, for the evaluation of the first reduction time (?), death time, and microbiological shelf life (the break-point was set to 7 log cfu/mL). Bifidobacterium animalis subsp. lactis experienced the highest ?-value (23.21 d) and death time (96.59 d) in the red-fruit juice at 4 °C, whereas L. plantarum was the most promising strain in apple juice at 37 °C. Biocitro and lemon extract did not exert a biocidal effect toward probiotics; moreover, the probiotics controlled the growth of Z. bailii and the combination of L. plantarum with 40 ppm of biocitro reduced the level of the yeast after 18 d by 2 log cfu/mL. PMID:24245895

  4. Identification of ecotype-specific marker genes for categorization of beer-spoiling Lactobacillus brevis.

    PubMed

    Behr, Jürgen; Geissler, Andreas J; Preissler, Patrick; Ehrenreich, Armin; Angelov, Angel; Vogel, Rudi F

    2015-10-01

    The tolerance to hop compounds, which is mainly associated with inhibition of bacterial growth in beer, is a multi-factorial trait. Any approaches to predict the physiological differences between beer-spoiling and non-spoiling strains on the basis of a single marker gene are limited. We identified ecotype-specific genes related to the ability to grow in Pilsner beer via comparative genome sequencing. The genome sequences of four different strains of Lactobacillus brevis were compared, including newly established genomes of two highly hop tolerant beer isolates, one strain isolated from faeces and one published genome of a silage isolate. Gene fragments exclusively occurring in beer-spoiling strains as well as sequences only occurring in non-spoiling strains were identified. Comparative genomic arrays were established and hybridized with a set of L. brevis strains, which are characterized by their ability to spoil beer. As result, a set of 33 and 4 oligonucleotide probes could be established specifically detecting beer-spoilers and non-spoilers, respectively. The detection of more than one of these marker sequences according to a genetic barcode enables scoring of L. brevis for their beer-spoiling potential and can thus assist in risk evaluation in brewing industry. PMID:26187837

  5. Purification and characterization of bacteriocin produced by oral Lactobacillus paracasei SD1.

    PubMed

    Wannun, P; Piwat, S; Teanpaisan, R

    2014-06-01

    The present study aimed to purify and characterize the antimicrobial protein from Lactobacillus paracasei SD1, which is a strain from the human oral cavity. Antimicrobial activity was obtained from purifying the culture supernatant of L. paracasei SD1. Purification of the active compound was achieved with ammonium sulfate precipitation followed by chloroform and gel filtration chromatography. As revealed by SDS-PAGE, the active fraction was homogeneous, showing a protein with an approximate molecular weight of 25,000 Da. It was confirmed as having a molecular mass of 24,028.2 Da by mass spectrometry. The antimicrobial compound, named "paracasin SD1", exhibited a broad spectrum against oral pathogens. Paracasin SD1 was stable in a pH range between 3.0 and 8.0 at 100 °C for 5 min, and showed resistance to ?-amylase, catalase, lysozyme and whole saliva. However, its activity was lost after proteinase K and trypsin treatment. The results obtained suggest the possibility of using paracasin SD1 for application in prevention/treatment of oral diseases. PMID:24636984

  6. Production and partial characterization of exopolysaccharides produced by two Lactobacillus suebicus strains isolated from cider.

    PubMed

    Ibarburu, Idoia; Puertas, Ana Isabel; Berregi, Iñaki; Rodríguez-Carvajal, Miguel A; Prieto, Alicia; Dueñas, M Teresa

    2015-12-01

    Many lactic acid bacteria synthesize extracellular polysaccharides (exopolysaccharides, EPSs) with a large variation in structure and potential functional properties. Although EPS production can produce detrimental effects in alcoholic beverages, these polymers play an important role in the rheological behavior and texture of fermented products. In this work, EPS production by two Lactobacillus suebicus strains, which were isolated from ropy ciders, was examined in a semidefined medium. The existence of priming glycosyltransferase encoding genes was detected by PCR. In addition, the preliminary characterization of the polymers was undertaken. Molecular masses were determined by size exclusion chromatography revealing the presence of two peaks, corresponding to polymers of high- and low-molecular-weight in all fractions. The composition of the EPS fractions was analyzed by gas chromatography-mass spectrometry after acid hydrolysis, revealing that they contained glucose, galactose, N-acetylglucosamine and phosphate, although in different ratios, suggesting that a mixture of polysaccharides is being synthesized. We also examined the influence of the sugar source (glucose, ribose, xylose, or arabinose) and pH conditions on growth and EPS production. PMID:26241490

  7. Structural investigation of cell wall polysaccharides of Lactobacillus delbrueckii subsp. bulgaricus 17.

    PubMed

    Vinogradov, E; Sadovskaya, I; Cornelissen, A; van Sinderen, D

    2015-09-01

    Lactobacilli are valuable strains for commercial (functional) food fermentations. Their cell surface-associated polysaccharides (sPSs) possess important functional properties, such as acting as receptors for bacteriophages (bacterial viruses), influencing autolytic characteristics and providing protection against antimicrobial peptides. The current report provides an elaborate molecular description of several surface carbohydrates of Lactobacillus delbrueckii subsp. bulgaricus strain 17. The cell surface of this strain was shown to contain short chain poly(glycerophosphate) teichoic acids and at least two different sPSs, designated here as sPS1 and sPS2, whose chemical structures were examined by 2D nuclear magnetic resonance spectroscopy and methylation analysis. Neutral branched sPS1, extracted with n-butanol, was shown to be composed of hexasaccharide repeating units (-[?-d-Glcp-(1-3)-]-4-?-l-Rhap2OAc-4-?-d-Glcp-[?-d-Galp-(1-3)]-4-?-Rhap-3-?-d-Galp-), while the major component of the TCA-extracted sPS2 was demonstrated to be a linear d-galactan with the repeating unit structure being (-[Gro-3P-(1-6)-]-3-?-Galf-3-?-Galp-2-?-Galf-6-?-Galf-3-?-Galp-). PMID:26117227

  8. In Lactobacillus pentosus, the olive brine adaptation genes are required for biofilm formation.

    PubMed

    Perpetuini, G; Pham-Hoang, B N; Scornec, H; Tofalo, R; Schirone, M; Suzzi, G; Cavin, J F; Waché, Y; Corsetti, A; Licandro-Seraut, H

    2016-01-01

    Lactobacillus pentosus is one of the few lactic acid bacteria (LAB) species capable of surviving in olive brine, and thus desirable during table olive fermentation. We have recently generated mutants of the efficient strain L. pentosus C11 by transposon mutagenesis and identified five mutants unable to survive and adapt to olive brine conditions. Since biofilm formation represents one of the main bacterial strategy to survive in stressful environments, in this study, the capacity of adhesion and formation of biofilm on olive skin was investigated for this strain and five derivative mutants which are interrupted in metabolic genes (enoA1 and gpi), and in genes of unknown function ("oba" genes). Confocal microscopy together with bacteria count revealed that the sessile state represented the prevailing L. pentosus C11 life-style during table olive fermentation. The characterization of cell surface properties showed that mutants present less hydrophobic and basic properties than the wild type (WT). In fact, their ability to adhere to both abiotic (polystyrene plates) and biotic (olive skin) surfaces was lower than that of the WT. Confocal microscopy revealed that mutants adhered sparsely to the olive skin instead of building a thin, multilayer biofilm. Moreover, RT-qPCR showed that the three genes enoA1, gpi and obaC were upregulated in the olive biofilm compared to the planktonic state. Thus enoA1, gpi and "oba" genes are necessary in L. pentosus to form an organized biofilm on the olive skin. PMID:26447789

  9. Effect of Palm or Coconut Solid Lipid Nanoparticles (SLNs) on Growth of Lactobacillus plantarum in Milk

    PubMed Central

    Jo, Yeon-Ji; Choi, Mi-Jung

    2015-01-01

    This study was performed to investigate the effect of palm or coconut solid lipid nanoparticles (PO-SLNs or CO-SLNs) on growth of Lactobacillus plantarum (L. plantarum) in milk during storage period. The PO or CO (0.1% or 1.0%) was dispersed both in distilled water (DW) and ultra high temperature milk (UHTM), and subsequently emulsified with Tween® 80 by ultrasonication (30% power, 2 min). Increase in particle size and encapsulation efficiency (EE%) in DW was observed with an increase in oil concentration, whereas a decrease in ?-potential of SLNs was noted with an increment in oil concentration. Moreover, the CO-SLNs exhibited relatively smaller particle size and higher EE% than PO-SLNs. The CO-SLNs were found to be more stable than PO-SLNs. Higher lipid oxidation of PO or CO-SLNs in UHTM was observed during the storage test, when compared to PO or CO-SLNs in DW. However, there was no remarkable difference in lipid oxidation during storage period (p>0.05). In the growth test, the viability of L. plantarum in control (without PO or CO-SLNs in DW) exhibited a dramatic decrease with increasing storage period. In addition, viability of L. plantarum of PO or CO-SLNs in UHTM was higher than that of SLNs in DW. Based on the present study, production of SLNs containing PO or CO in UHTM is proposed, which can be used in lactobacilli fortified beverages in food industry.

  10. In vitro evaluation of the probiotic potential of Lactobacillus salivarius SMXD51.

    PubMed

    Messaoudi, S; Madi, A; Prévost, H; Feuilloley, M; Manai, M; Dousset, X; Connil, N

    2012-12-01

    Lactobacillus salivarius SMXD51 was previously isolated from the cecum of a Tunisian poultry and found to produce a bacteriocin-like substance highly active against the foodborne pathogen Campylobacter jejuni. The aim of this study was to examine some probiotic properties of the strain: acid and bile tolerance, capacity of adhesion, stimulation of immune defences (IL-6, IL-8, IL-10 and ?-defensin 2), and modulation of the barrier integrity. The results showed that L. salivarius SMXD51 can tolerate gastrointestinal conditions (acid and bile), adhere to intestinal cells and stimulate the immune system. The bacterium strengthened the intestinal barrier functions through the increase of the transepithelial electrical resistance (TEER) and reinforcement of the F-actin cytoskeleton. One hour pretreatment with L. salivarius SMXD51 protected against Pseudomonas aeruginosa PAO1-induced decrease of TEER and damage of the F-actin cytoskeleton. Our results highlight that L. salivarius SMXD51 fulfils the principle requirements of an efficient probiotic and may be seen as a reliable candidate for further validation studies in chicken. PMID:23122647

  11. Genome –Scale Reconstruction of Metabolic Networks of Lactobacillus casei ATCC 334 and 12A

    PubMed Central

    Vinay-Lara, Elena; Hamilton, Joshua J.; Stahl, Buffy; Broadbent, Jeff R.; Reed, Jennifer L.; Steele, James L.

    2014-01-01

    Lactobacillus casei strains are widely used in industry and the utility of this organism in these industrial applications is strain dependent. Hence, tools capable of predicting strain specific phenotypes would have utility in the selection of strains for specific industrial processes. Genome-scale metabolic models can be utilized to better understand genotype-phenotype relationships and to compare different organisms. To assist in the selection and development of strains with enhanced industrial utility, genome-scale models for L. casei ATCC 334, a well characterized strain, and strain 12A, a corn silage isolate, were constructed. Draft models were generated from RAST genome annotations using the Model SEED database and refined by evaluating ATP generating cycles, mass-and-charge-balances of reactions, and growth phenotypes. After the validation process was finished, we compared the metabolic networks of these two strains to identify metabolic, genetic and ortholog differences that may lead to different phenotypic behaviors. We conclude that the metabolic capabilities of the two networks are highly similar. The L. casei ATCC 334 model accounts for 1,040 reactions, 959 metabolites and 548 genes, while the L. casei 12A model accounts for 1,076 reactions, 979 metabolites and 640 genes. The developed L. casei ATCC 334 and 12A metabolic models will enable better understanding of the physiology of these organisms and be valuable tools in the development and selection of strains with enhanced utility in a variety of industrial applications. PMID:25365062

  12. Hydrolysis of Oleuropein by Lactobacillus plantarum Strains Associated with Olive Fermentation

    PubMed Central

    Ciafardini, G.; Marsilio, V.; Lanza, B.; Pozzi, N.

    1994-01-01

    Oleuropein (Chemical Abstracts Service registry number 32619-42-4), a bitter-tasting secoiridoid glucoside commonly found in leaves of the olive tree as well as in olives (Olea europaea L.), was found to be hydrolyzed by the ?-glucosidase (EC 3.2.1.2.1) produced by oleuropeinolytic Lactobacillus plantarum-type strains. Three strains, designated B17, B20, and B21, were isolated from the brine of naturally ripe olives not treated with alkali. These strains were rod-shaped forms, grown at a pH 3.5 limit, and tolerated 1% oleuropein and 8% NaCl in the growth medium. The ?-glucosidase produced hydrolyzed 5-bromo-4-chloro-3-indolyl-?-d-glucopy-ranoside as well as oleuropein. The presence of 2% glucose in the medium inhibited activity by 40 to 50%, depending on the bacterial strain. Chromatographic analysis of the trimethylsilyl derivatives of the products obtained after 7 days of incubation at 30°C of strain B21 showed all the hydrolysis products of oleuropein, i.e., aglycone, iridoid monoterpen, and 3,4-dihydroxyphenylethanol (hydroxytyrosol). Oleuropein and its aglycone after 21 days of incubation decreased to trace levels with the simultaneous increase in concentration of ?-3,4-dihydroxyphenylethanol. Images PMID:16349442

  13. Enhanced antiinflammatory capacity of a Lactobacillus plantarum mutant synthesizing modified teichoic acids.

    PubMed

    Grangette, Corinne; Nutten, Sophie; Palumbo, Emmanuelle; Morath, Siegfried; Hermann, Corinna; Dewulf, Joelle; Pot, Bruno; Hartung, Thomas; Hols, Pascal; Mercenier, Annick

    2005-07-19

    Teichoic acids (TAs), and especially lipoteichoic acids (LTAs), are one of the main immunostimulatory components of pathogenic Gram-positive bacteria. Their contribution to the immunomodulatory properties of commensal bacteria and especially of lactic acid bacteria has not yet been investigated in detail. To evaluate the role of TAs in the interaction between lactic acid bacteria and the immune system, we analyzed the antiinflammatory properties of a mutant of Lactobacillus plantarum NCIMB8826 affected in the TA biosynthesis pathway both in vitro (mononuclear cells stimulation) and in vivo (murine model of colitis). This Dlt- mutant was found to incorporate much less D-Ala in its TAs than the WT strain. This defect significantly impacted the immunomodulation reactions induced by the bacterium, as shown by a dramatically reduced secretion of proinflammatory cytokines by peripheral blood mononuclear cells and monocytes stimulated by the Dlt- mutant as compared with the parental strain. Concomitantly, a significant increase in IL-10 production was stimulated by the Dlt- mutant in comparison with the WT strain. Moreover, the proinflammatory capacity of L. plantarum-purified LTA was found to be Toll-like receptor 2-dependent. Consistent with the in vitro results, the Dlt- mutant was significantly more protective in a murine colitis model than its WT counterpart. The results indicated that composition of LTA within the whole-cell context of L. plantarum can modulate proinflammatory or antiinflammatory immune responses. PMID:15985548

  14. Enhanced antiinflammatory capacity of a Lactobacillus plantarum mutant synthesizing modified teichoic acids

    PubMed Central

    Grangette, Corinne; Nutten, Sophie; Palumbo, Emmanuelle; Morath, Siegfried; Hermann, Corinna; Dewulf, Joelle; Pot, Bruno; Hartung, Thomas; Hols, Pascal; Mercenier, Annick

    2005-01-01

    Teichoic acids (TAs), and especially lipoteichoic acids (LTAs), are one of the main immunostimulatory components of pathogenic Gram-positive bacteria. Their contribution to the immunomodulatory properties of commensal bacteria and especially of lactic acid bacteria has not yet been investigated in detail. To evaluate the role of TAs in the interaction between lactic acid bacteria and the immune system, we analyzed the antiinflammatory properties of a mutant of Lactobacillus plantarum NCIMB8826 affected in the TA biosynthesis pathway both in vitro (mononuclear cells stimulation) and in vivo (murine model of colitis). This Dlt- mutant was found to incorporate much less d-Ala in its TAs than the WT strain. This defect significantly impacted the immunomodulation reactions induced by the bacterium, as shown by a dramatically reduced secretion of proinflammatory cytokines by peripheral blood mononuclear cells and monocytes stimulated by the Dlt- mutant as compared with the parental strain. Concomitantly, a significant increase in IL-10 production was stimulated by the Dlt- mutant in comparison with the WT strain. Moreover, the proinflammatory capacity of L. plantarum-purified LTA was found to be Toll-like receptor 2-dependent. Consistent with the in vitro results, the Dlt- mutant was significantly more protective in a murine colitis model than its WT counterpart. The results indicated that composition of LTA within the whole-cell context of L. plantarum can modulate proinflammatory or antiinflammatory immune responses. PMID:15985548

  15. Lactobacillus rhamnosus GG Lysate Increases Re-Epithelialization of Keratinocyte Scratch Assays by Promoting Migration.

    PubMed

    Mohammedsaeed, Walaa; Cruickshank, Sheena; McBain, Andrew J; O'Neill, Catherine A

    2015-01-01

    A limited number of studies have investigated the potential of probiotics to promote wound healing in the digestive tract. The aim of the current investigation was to determine whether probiotic bacteria or their extracts could be beneficial in cutaneous wound healing. A keratinocyte monolayer scratch assay was used to assess re-epithelialization; which comprises keratinocyte proliferation and migration. Primary human keratinocyte monolayers were scratched then exposed to lysates of Lactobacillus (L) rhamnosus GG, L. reuteri, L. plantarum or L. fermentum. Re-epithelialization of treated monolayers was compared to that of untreated controls. Lysates of L. rhamnosus GG and L. reuteri significantly increased the rate of re-epithelialization, with L. rhamnosus GG being the most efficacious. L. reuteri increased keratinocyte proliferation while L. rhamnosus GG lysate significantly increased proliferation and migration. Microarray analysis of L. rhamnosus GG treated scratches showed increased expression of multiple genes including the chemokine CXCL2 and its receptor CXCR2. These are involved in normal wound healing where they stimulate keratinocyte proliferation and/or migration. Increased protein expression of both CXCL2 and CXCR2 were confirmed by ELISA and immunoblotting. These data demonstrate that L. rhamnosus GG lysate accelerates re-epithelialization of keratinocyte scratch assays, potentially via chemokine receptor pairs that induce keratinocyte migration. PMID:26537246

  16. Lactobacillus plantarum TN8 exhibits protective effects on lipid, hepatic and renal profiles in obese rat.

    PubMed

    Ben Salah, Riadh; Trabelsi, Imen; Hamden, Khaled; Chouayekh, Hichem; Bejar, Samir

    2013-10-01

    This study aimed to first investigate the immuno-modulatory effects of six newly isolated lactic acid bacteria (LAB) on the peripheral blood mononuclear cells (PBMC) of Wistar rats. Except for Lactobacillus plantarum TN8, all the other strains were noted to induce high levels of pro-inflammatory cytokine IL-12 and low levels of anti-inflammatory cytokine IL-10. The strains also generated low ratios of IL-10/IL-12 cytokine. Strain TN8 was, on the other hand, noted to induce an increase in anti-inflammatory IL-10 cytokine secretion rates and a decrease in pro-inflammatory IL-12, IFN-? and TNF-? cytokine production. The oral administration of TN8 improved the hepatic and urinary functions of obese rats by inducing decreases (P < 0.05) in alanine amino transferase (ALAT), gamma glutamyl transferase (GGT), plasmatic triglycerides, total cholesterol concentrations, creatinine, urea, and body weight when compared to the control group of animals that underwent an increase in aspartate amino transferase (ASAT) and high density lipoprotein (HDL). Overall, the findings indicate that strain TN8 exhibited a number of attractive properties that might open new promising opportunities for the improvement of various parameters related to animal health performance and the avoidance of antibiotics and drugs as promoting factors. PMID:23891961

  17. Lactobacillus casei reduces susceptibility to type 2 diabetes via microbiota-mediated body chloride ion influx

    PubMed Central

    Zhang, Yong; Guo, Xiao; Guo, Jianlin; He, Qiuwen; Li, He; Song, Yuqin; Zhang, Heping

    2014-01-01

    Gut microbiota mediated low-grade inflammation is involved in the onset of type 2 diabetes (T2DM). In this study, we used a high fat sucrose (HFS) diet-induced pre-insulin resistance and a low dose-STZ HFS rat models to study the effect and mechanism of Lactobacillus casei Zhang in protecting against T2DM onset. Hyperglycemia was favorably suppressed by L. casei Zhang treatment. Moreover, the hyperglycemia was connected with type 1 immune response, high plasma bile acids and urine chloride ion loss. This chloride ion loss was significantly prevented by L. casei via upregulating of chloride ion-dependent genes (ClC1-7, GlyR?1, SLC26A3, SLC26A6, GABAA?1, Bestrophin-3 and CFTR). A shift in the caecal microflora, particularly the reduction of bile acid 7?-dehydroxylating bacteria, and fecal bile acid profiles also occurred. These change coincided with organ chloride influx. Thus, we postulate that the prevention of T2DM onset by L. casei Zhang may be via a microbiota-based bile acid-chloride exchange mechanism. PMID:25133590

  18. Influence of cooling temperature and duration on cold adaptation of Lactobacillus acidophilus RD758.

    PubMed

    Wang, Yu; Delettre, Jérôme; Guillot, Alain; Corrieu, Georges; Béal, Catherine

    2005-06-01

    The effect of different cooling temperatures and durations on resistance to freezing and to frozen storage at -20 degrees C in Lactobacillus acidophilus RD758 was studied, by using a central composite rotatable design. A cold adaptation was observed when the cells were maintained at moderate temperature (26 degrees C) for a long time (8h) before being cooled to the final temperature of 15 degrees C. These conditions led to a low rate of loss in acidification activity during frozen storage (0.64 minday(-1)) and a high residual acidification activity after 180 days of frozen storage (1011 min). The experimental design allowed us to determine optimal cooling conditions, which were established at 28 degrees C during 8h. Adaptation to cold temperatures was related to an increase in the unsaturated to saturated fatty acid ratio and in the relative cycC19:0 fatty acid concentration. Moreover, an increased synthesis of four specific proteins was observed as an adaptive response to the optimal cooling conditions. They included the stress protein ATP-dependent ClpP and two cold induced proteins: pyruvate kinase and a putative glycoprotein endopeptidase. PMID:15925581

  19. Physicochemical properties of a water soluble extracellular homopolysaccharide from Lactobacillus reuteri SK24.003.

    PubMed

    Miao, Ming; Ma, Yajun; Huang, Chao; Jiang, Bo; Cui, Steve W; Zhang, Tao

    2015-10-20

    A water soluble exopolysaccharide was obtained from Lactobacillus reuteri SK24.003 and its rheological and structural properties were investigated. The polysaccharide yield reached around 40.8g/L for 48h of fermentation. The purified exopolysaccharide was an ?-d-glucan with the molecular weight of 2.50×10(7)g/mol. Thermal analysis showed the exopolysaccharide broke down above 200°C, the weight dramatically lost (around 53.0%) from 230 to 340°C and the melting point was 147.7°C with the enthalpy value of 78.4J/g. The glucan solution (3.0-9.0%, v/w) has an opalescent, milky-white color and the concentration affected the depth of color. The viscosity increased rapidly with concentration in the range of 3.0-20% (w/v) with relatively high viscosity as well as poor solubility at a concentration higher than 20% (w/v). The solution also exhibited the non-Newtonian pseudoplastic behavior with increasing shear rate from 0.01 to 100l/s. This glucan had a high viscosity in solution and this could make it suitable for usage as gum substitute in some processing food. PMID:26256197

  20. Quantitative Proteogenomics and the Reconstruction of the Metabolic Pathway in Lactobacillus mucosae LM1

    PubMed Central

    Lee, Ji-Yoon

    2015-01-01

    Lactobacillus mucosae is a natural resident of the gastrointestinal tract of humans and animals and a potential probiotic bacterium. To understand the global protein expression profile and metabolic features of L. mucosae LM1 in the early stationary phase, the QExactiveTM Hybrid Quadrupole-Orbitrap Mass Spectrometer was used. Characterization of the intracellular proteome identified 842 proteins, accounting for approximately 35% of the 2,404 protein-coding sequences in the complete genome of L. mucosae LM1. Proteome quantification using QExactiveTM Orbitrap MS detected 19 highly abundant proteins (> 1.0% of the intracellular proteome), including CysK (cysteine synthase, 5.41%) and EF-Tu (elongation factor Tu, 4.91%), which are involved in cell survival against environmental stresses. Metabolic pathway annotation of LM1 proteome using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database showed that half of the proteins expressed are important for basic metabolic and biosynthetic processes, and the other half might be structurally important or involved in basic cellular processes. In addition, glycogen biosynthesis was activated in the early stationary phase, which is important for energy storage and maintenance. The proteogenomic data presented in this study provide a suitable reference to understand the protein expression pattern of lactobacilli in standard conditions.

  1. Influence of gastrointestinal stress on autoinducer-2 activity of two Lactobacillus species.

    PubMed

    Yeo, Soyoung; Park, Hyunjoon; Ji, Yosep; Park, Soyoung; Yang, Jaesik; Lee, Jieun; Mathara, Julius M; Shin, Heuynkil; Holzapfel, Wilhelm

    2015-07-01

    Quorum sensing is a bacterial communication signalling system that regulates the expression of certain target genes with autoinducers in a cell density-dependent manner. The universal luxS-mediated quorum sensing using the autoinducer-2 (AI-2) signal is present in a wide variety of bacteria with only sparse information on probiotic lactobacilli. Effective probiotics should exhibit tolerance and adaptation to stress conditions typical of the GIT. Adhesion to human intestinal epithelial cells and competitive exclusion of pathogens are also considered important. The AI-2 signal system plays an important role in the response of probiotic lactobacilli to the surrounding environment. Intraspecies-related changes in quorum signalling in the GIT were determined by monitoring the AI-2 activity of two strains each of Lactobacillus rhamnosus and L. plantarum under various stress conditions. Modulation of the AI-2 activity of all the strains was induced by stress responses to pH, bile acid, temperature, osmotic pressure and starvation, and was both species- and strain-specific. AI-2 inhibition correlated with a reduction in the stress-related genes of L. rhamnosus. We therefore suggest that AI-2 quorum signalling of probiotic lactobacilli may represent one way of adapting to the host's ecosystem and of interacting within the intestinal environment. PMID:26092949

  2. Lactobacillus acidophilus NCFM affects vitamin E acetate metabolism and intestinal bile acid signature in monocolonized mice

    PubMed Central

    Roager, Henrik M; Sulek, Karolina; Skov, Kasper; Frandsen, Henrik L; Smedsgaard, Jørn; Wilcks, Andrea; Skov, Thomas H; Villas-Boas, Silas G; Licht, Tine R

    2014-01-01

    Monocolonization of germ-free (GF) mice enables the study of specific bacterial species in vivo. Lactobacillus acidophilus NCFMTM (NCFM) is a probiotic strain; however, many of the mechanisms behind its health-promoting effect remain unknown. Here, we studied the effects of NCFM on the metabolome of jejunum, cecum, and colon of NCFM monocolonized (MC) and GF mice using liquid chromatography coupled to mass-spectrometry (LC-MS). The study adds to existing evidence that NCFM in vivo affects the bile acid signature of mice, in particular by deconjugation. Furthermore, we confirmed that carbohydrate metabolism is affected by NCFM in the mouse intestine as especially the digestion of oligosaccharides (penta- and tetrasaccharides) was increased in MC mice. Additionally, levels of ?-tocopherol acetate (vitamin E acetate) were higher in the intestine of GF mice than in MC mice, suggesting that NCFM affects the vitamin E acetate metabolism. NCFM did not digest vitamin E acetate in vitro, suggesting that direct bacterial metabolism was not the cause of the altered metabolome in vivo. Taken together, our results suggest that NCFM affects intestinal carbohydrate metabolism, bile acid metabolism and vitamin E metabolism, although it remains to be investigated whether this effect is unique to NCFM. PMID:24717228

  3. Application of response surface methodology to enhancement of biomass production by Lactobacillus rhamnosus E/N

    PubMed Central

    Polak-Berecka, Magdalena; Wa?ko, Adam; Kordowska-Wiater, Monika; Targo?ski, Zdzis?aw; Kubik-Komar, Agnieszka

    2011-01-01

    Response surface methodology (RSM) was employed to study the effects of various medium components on biomass production by Lactobacillus rhamnosus E/N. This strain is commonly used in the pharmaceutical and food industries due to its beneficial effect on the human gut and general health. The best medium composition derived from RSM regression was (in g/l) glucose 15.44, sodium pyruvate 3.92, meat extract 8.0, potassium phosphate 1.88, sodium acetate 4.7, and ammonium citrate 1.88. With this medium composition biomass production was 23 g/l of dry cell weight after 18 h of cultivation in bioreactor conditions, whereas on MRS the yield of biomass was 21 g/l of dry cell weight. The cost of 1 g of biomass obtained on MRS broth was calculated at the level of 0.44 € whereas on the new optimal medium it was 25% lower. It may be concluded then, that the new medium, being cheaper than the control MRS allows large scale commercial cultivation of the L. rhamnosus strain. This study is of relevance to food industry because the possibility to obtain high yield of bacterial biomass is necessary step in manufacturing of probiotic food. PMID:24031782

  4. Purification and characterization of plantaricin LR14: a novel bacteriocin produced by Lactobacillus plantarum LR/14.

    PubMed

    Tiwari, Santosh Kumar; Srivastava, Sheela

    2008-07-01

    Bacteriocin produced by Lactobacillus plantarum strain LR/14 was purified to homogeneity by a multi-step protocol consisting of ammonium sulfate precipitation, cation-exchange chromatography, gel-filtration, and reverse-phase FPLC. L. plantarum LR/14 secreted a low-molecular-weight bacteriocin consisting of two peptides designated as plantaricin LR14alpha and -beta with molecular mass of 3,012.46 and 5,605.74 Da, respectively. The purified peptides were characterized to be highly thermostable and active in acidic pH range, with a pI of >10.0. Both alpha and beta peptides showed bactericidal mode of action against indicator strain, Micrococcus luteus and together showed a synergistic action. These peptides were differentially sensitive to a range of proteolytic enzymes, indicating differences in their composition. Amino acid sequencing revealed that the N-terminus in both the cases is blocked; thus, only a partial sequence could be obtained after CNBr digestion. These sequences, when compared with those available in the database, showed no homology with known bacteriocins, indicating it to be a novel compound. PMID:18496687

  5. Purification and Characterization of Plantaricin ZJ5, a New Bacteriocin Produced by Lactobacillus plantarum ZJ5

    PubMed Central

    Song, Da-Feng; Zhu, Mu-Yuan; Gu, Qing

    2014-01-01

    The aim of this study is to investigate the antimicrobial potential of Lactobacillus plantarum ZJ5, a strain isolated from fermented mustard with a broad range of inhibitory activity against both Gram-positive and Gram-negative bacteria. Here we present the peptide plantaricin ZJ5 (PZJ5), which is an extreme pH and heat-stable. However, it can be digested by pepsin and proteinase K. This peptide has strong activity against Staphylococcus aureus. PZJ5 has been purified using a multi-step process, including ammonium sulfate precipitation, cation-exchange chromatography, hydrophobic interactions and reverse-phase chromatography. The molecular mass of the peptide was found to be 2572.9 Da using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The primary structure of this peptide was determined using amino acid sequencing and DNA sequencing, and these analyses revealed that the DNA sequence translated as a 44-residue precursor containing a 22-amino-acid N-terminal extension that was of the double-glycine type. The bacteriocin sequence exhibited no homology with known bacteriocins when compared with those available in the database, indicating that it was a new class IId bacteriocin. PZJ5 from a food-borne strain may be useful as a promising probiotic candidate. PMID:25147943

  6. Molecular mechanisms behind the antimicrobial activity of hop iso-?-acids in Lactobacillus brevis.

    PubMed

    Schurr, Benjamin C; Hahne, Hannes; Kuster, Bernhard; Behr, Jürgen; Vogel, Rudi F

    2015-04-01

    The main bittering component in beer, hop iso-?-acids, have been characterised as weak acids, which act as ionophores impairing microbial cells' function under acidic conditions as present in beer. Besides medium pH, divalent cations play a central role regarding the efficacy of the antimicrobial effect. The iso-?-acids' non-bitter derivatives humulinic acids can be found in isomerised hop extracts and can be generated during hop storage. Therefore, they have been under investigation concerning their influence on beer sensory properties. This study sketches the molecular mechanism behind iso-?-acids' antimicrobial activity in Lactobacillus (L.) brevis regarding their ionophore activity versus the dependence of the inhibitory potential on manganese binding, and suggests humulinic acids as novel tasteless food preservatives. We designed and synthesised chemically modified iso-?-acids to enhance the basic understanding of the molecular mechanism of antimicrobial iso-?-acids. It could be observed that a manganese-binding dependent transmembrane redox reaction (oxidative stress) plays a crucial role in inhibition. Privation of an acidic hydroxyl group neither erased ionophore activity, nor did it entirely abolish antimicrobial activity. Humulinic acids proved to be highly inhibitory, even outperforming iso-?-acids. PMID:25475328

  7. Protective Effects of Lactobacillus plantarum CCFM8246 against Copper Toxicity in Mice

    PubMed Central

    Li, Xiaoxiao; Zhai, Qixiao; Wang, Gang; Zhang, Qiuxiang; Zhang, Hao; Chen, Wei

    2015-01-01

    Lactobacillus plantarum CCFM8246, which has a relatively strong copper binding capacity and tolerance to copper ions, was obtained by screening from 16 lactic acid bacteria in vitro. The selected strain was then applied to a mouse model to evaluate its protective function against copper intoxication in vivo. The experimental mice were divided into an intervention group and a therapy group; mice in the intervention group received co-administration of CCFM8246 and a copper ion solution by gavage, while mice in the therapy group were treated with CCFM8246 after 4 weeks of copper exposure. In both two groups, mice treated with copper alone and that treated with neither CCFM8246 nor copper served as positive and negative controls, respectively. At the end of the experimental period, the copper content in feces and tissues, the activity of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum, and oxidation stress indices in liver and kidney tissue were determined. Learning and memory ability was evaluated by Morris water maze experiments. The results indicated that treatment with CCFM8246 significantly increased the copper content in feces to promote copper excretion, reduce the accumulation of copper in tissues, reverse oxidative stress induced by copper exposure, recover the ALT and AST in serum and improve the spatial memory of mice. PMID:26605944

  8. Evaluation of resistance to low pH and bile salts of human Lactobacillus spp. isolates.

    PubMed

    Fuochi, Virginia; Petronio, Giulio Petronio; Lissandrello, Edmondo; Furneri, Pio Maria

    2015-09-01

    There are nearly 100 trillion bacteria in the intestine that together form the intestinal microbiota. They are 'good' bacteria because they help to maintain a physiological balance and are called probiotics. Probiotics must have some important characteristics: be safe for humans, be resistant to the low pH in the stomach, as well as bile salts and pancreatic juice. Indeed, their survival is the most important factor, so that they can arrive alive in the intestine and are able to form colonies, at least temporarily. The aim of our study was the evaluation of resistance of Lactobacillus isolates from fecal and oral swabs compared to that found in a commercial product. Seven strains were randomly chosen: L. jensenii, L. gasseri, L. salivarius, L. fermentum, L. rhamnosus, L. crispatus, and L. delbrueckii. We observed a large variability in the results: L. gasseri and L. fermentum were the most resistance to low pH, while only L. gasseri showed the best survival rate to bile salts. Interestingly, the commercial product did not show tolerance to both low pH and bile salts. PMID:26216909

  9. Production, purification and characterization of bacteriocin from Lactobacillus murinus AU06 and its broad antibacterial spectrum

    PubMed Central

    Elayaraja, Sivaramasamy; Annamalai, Neelamegam; Mayavu, Packiyam; Balasubramanian, Thangavel

    2014-01-01

    Objective To study the production, purification and characterization of bacteriocin from Lactobacillus murinus AU06 isolated from marine sediments and its broad spectrum of inhibition against fish pathogens. Methods The selected strain was used in production, purification and characterized of bacteriocin. In addition, purified bacteriocin was tested for its antimicrobial activity against fish pathogens. Results In the present study, the bacteriocin production was found to be higher at 35 °C, pH 6.0 and was purified to 4.74 fold with 55. 38 U/mg of specific activity with the yield of 28.92%. The molecular weight of the purified bacteriocin was estimated as 21 kDa. The purified bacteriocin exhibited complete inactivation of antimicrobial activity when treated with proteinase K, pronase, chymotrypsin, trypsin, pepsin and papain. The purified bacteriocin exhibited broad inhibitory spectrum against both Gram positive and negative bacteria. Conclusions It is concluded that the ability of bacteriocin in inhibiting a wide-range of pathogenic bacteria is of potential interest for food safety and may have future applications in food preservative. PMID:25183102

  10. Flow Cytometric Testing of Green Fluorescent Protein-Tagged Lactobacillus rhamnosus GG for Response to Defensins

    PubMed Central

    De Keersmaecker, Sigrid C. J.; Braeken, Kristien; Verhoeven, Tine L. A.; Perea Vélez, Mónica; Lebeer, Sarah; Vanderleyden, Jos; Hols, Pascal

    2006-01-01

    Lactobacillus rhamnosus GG is of general interest as a probiotic. Although L. rhamnosus GG is often used in clinical trials, there are few genetic tools to further determine its mode of action or to develop it as a vehicle for heterologous gene expression in therapy. Therefore, we developed a reproducible, efficient electroporation procedure for L. rhamnosus GG. The best transformation efficiency obtained was 104 transformants per ?g of DNA. We validated this protocol by tagging L. rhamnosus GG with green fluorescent protein (GFP) using the nisin-controlled expression (NICE) system. Parameters for overexpression were optimized, which allowed expression of gfp in L. rhamnosus GG upon induction with nisin. The GFP+ strain can be used to monitor the survival and behavior of L. rhamnosus GG in vivo. Moreover, implementation of the NICE system as a gene expression switch in L. rhamnosus GG opens up possibilities for improving and expanding the performance of this strain. The GFP-labeled strain was used to demonstrate that L. rhamnosus GG is sensitive to human beta-defensin-2 but not to human beta-defensin-1. PMID:16820489

  11. Contribution of volatiles to the antifungal effect of Lactobacillus paracasei in defined medium and yogurt.

    PubMed

    Aunsbjerg, S D; Honoré, A H; Marcussen, J; Ebrahimi, P; Vogensen, F K; Benfeldt, C; Skov, T; Knøchel, S

    2015-02-01

    Lactic acid bacteria with antifungal properties can be used to control spoilage of food and feed. Previously, most of the identified metabolites have been isolated from cell-free fermentate of lactic acid bacteria with methods suboptimal for detecting possible contribution from volatiles to the antifungal activity. The role of volatile compounds in the antifungal activity of Lactobacillus paracasei DGCC 2132 in a chemically defined interaction medium (CDIM) and yogurt was therefore investigated with a sampling technique minimizing volatile loss. Diacetyl was identified as the major volatile produced by L. paracasei DGCC 2132 in CDIM. When the strain was added to a yogurt medium diacetyl as well as other volatiles also increased but the metabolome was more complex. Removal of L. paracasei DGCC 2132 cells from CDIM fermentate resulted in loss of both volatiles, including diacetyl, and the antifungal activity towards two strains of Penicillium spp. When adding diacetyl to CDIM or yogurt without L. paracasei DGCC 2132, marked inhibition was observed. Besides diacetyl, the antifungal properties of acetoin were examined, but no antifungal activity was observed. Overall, the results demonstrate the contribution of diacetyl in the antifungal effect of L. paracasei DGCC 2132 and indicate that the importance of volatiles may have been previously underestimated. PMID:25461608

  12. Combined effect of Lactobacillus acidophilus and ?-cyclodextrin on serum cholesterol in pigs.

    PubMed

    Alonso, L; Fontecha, J; Cuesta, P

    2016-01-01

    A total of twenty-four Yorkshire gilt pigs of 6-7 weeks of age were used in a 2×2 factorial experiment to determine the individual and combined effects of the inclusion of two dietary factors (cholesterol rich, 3 % ?-cyclodextrin (BCD) and Lactobacillus acidophilus cultures) on total cholesterol and LDL-cholesterol levels in blood serum. Pigs were assigned randomly to treatment groups (n 6). Total serum cholesterol concentrations decreased after 3 weeks in all the experimental treatment groups, including diets with BCD, L. acidophilus or both. Similar trends were observed for serum LDL-cholesterol concentrations among the experimental treatments. No statistically significant differences from the control group were observed in either total serum cholesterol or LDL-cholesterol concentrations (P<0·05) for each of the individual treatment groups: BCD or L. acidophilus. However, significant differences in total serum cholesterol concentrations were observed when comparing the combined treatment group (BCD and L. acidophilus) with the control group, which consisted of a basal diet and sterile milk. The combined treatment group exhibited 17·9 % lower total serum cholesterol concentration after 3 weeks. Similar significant differences were observed when comparing the combined effect experimental group with the control group after 3 weeks. The combined treatment group exhibited 27·9 % lower serum LDL-cholesterol concentrations. PMID:26467089

  13. Evidence for a Plasmid-Linked Restriction-Modification System in Lactobacillus helveticus

    PubMed Central

    de los Reyes-Gavilán, Clara G.; Limsowtin, Gaëtan K. Y.; Séchaud, Laurent; Veaux, Monique; Accolas, Jean-Pierre

    1990-01-01

    The presence of a restriction-modification (R/M) system against two bacteriophages, 328-B1 and hv, was demonstrated in three Lactobacillus helveticus strains, CNRZ 1094, CNRZ 1095, and CNRZ 1096. In addition, the burst size of phage 328-B1 in the three restrictive strains CNRZ 1094, CNRZ 1095, and CNRZ 1096 was reduced with respect to the values obtained in its propagating strain, CNRZ 328. Heating at 60°C did not inactivate the R/M system. Nonrestrictive variants from CNRZ 1094 were easily obtained under several culture conditions, but treatment with novobiocin at 42°C followed by storage at ?20°C resulted in drastic elimination of the R+/M+ phenotype from all clones tested. Electrophoretic analysis of CNRZ 1094 nonrestrictive variants revealed the concomitant loss of a 34-kb plasmid. Four EcoRI fragments from the 34-kb plasmid were cloned in the Escherichia coli vector pACYC184. The use of one or several of these fragments as probes confirmed the plasmidic location of the genes responsible for the R/M system. These probes also showed the presence of R/M plasmids in the two other restrictive strains, CNRZ 1095 and CNRZ 1096. Lactose-fermenting ability and/or proteolytic capacity was not linked to the 34-kb plasmid. Images PMID:16348347

  14. A Lactobacillus plantarum Esterase Active on a Broad Range of Phenolic Esters

    PubMed Central

    Esteban-Torres, María; Landete, José María; Reverón, Inés; Santamaría, Laura; de las Rivas, Blanca

    2015-01-01

    Lactobacillus plantarum is the lactic acid bacterial species most frequently found in the fermentation of food products of plant origin on which phenolic compounds are abundant. L. plantarum strains showed great flexibility in their ability to adapt to different environments and growth substrates. Of 28 L. plantarum strains analyzed, only cultures from 7 strains were able to hydrolyze hydroxycinnamic esters, such as methyl ferulate or methyl caffeate. As revealed by PCR, only these seven strains possessed the est_1092 gene. When the est_1092 gene was introduced into L. plantarum WCFS1 or L. lactis MG1363, their cultures acquired the ability to degrade hydroxycinnamic esters. These results support the suggestion that Est_1092 is the enzyme responsible for the degradation of hydroxycinnamic esters on the L. plantarum strains analyzed. The Est_1092 protein was recombinantly produced and biochemically characterized. Surprisingly, Est_1092 was able to hydrolyze not only hydroxycinnamic esters, since all the phenolic esters assayed were hydrolyzed. Quantitative PCR experiments revealed that the expression of est_1092 was induced in the presence of methyl ferulate, an hydroxycinnamic ester, but was inhibited on methyl gallate, an hydroxybenzoic ester. As Est_1092 is an enzyme active on a broad range of phenolic esters, simultaneously possessing feruloyl esterase and tannase activities, its presence on some L. plantarum strains provides them with additional advantages to survive and grow on plant environments. PMID:25746986

  15. Glycosylation of Phenolic Compounds by the Site-Mutated ?-Galactosidase from Lactobacillus bulgaricus L3

    PubMed Central

    Lu, Lili; Xu, Lijuan; Guo, Yuchuan; Zhang, Dayu; Qi, Tingting; Jin, Lan; Gu, Guofeng; Xu, Li; Xiao, Min

    2015-01-01

    ?-Galactosidases can transfer the galactosyl from lactose or galactoside donors to various acceptors and thus are especially useful for the synthesis of important glycosides. However, these enzymes have limitations in the glycosylation of phenolic compounds that have many physiological functions. In this work, the ?-galactosidase from Lactobacillus bulgaricus L3 was subjected to site-saturation mutagenesis at the W980 residue. The recombinant pET-21b plasmid carrying the enzyme gene was used as the template for mutation. The mutant plasmids were transformed into Escherichia coli cells for screening. One recombinant mutant, W980F, exhibited increased yield of glycoside when using hydroquinone as the screening acceptor. The enzyme was purified and the effects of the mutation on enzyme properties were determined in detail. It showed improved transglycosylation activity on novel phenolic acceptors besides hydroquinone. The yields of the glycosides produced from phenol, hydroquinone, and catechol were increased by 7.6% to 53.1%. Moreover, it generated 32.3% glycosides from the pyrogallol that could not be glycosylated by the wild-type enzyme. Chemical structures of these glycoside products were further determined by MS and NMR analysis. Thus, a series of novel phenolic galactosides were achieved by ?-galactosidase for the first time. This was a breakthrough in the enzymatic galactosylation of the challenging phenolic compounds of great values. PMID:25803778

  16. Cracking Streptococcus thermophilus to stimulate the growth of the probiotic Lactobacillus casei in co-culture.

    PubMed

    Ma, Chengjie; Ma, Aimin; Gong, Guangyu; Liu, Zhenmin; Wu, Zhengjun; Guo, Benheng; Chen, Zhengjun

    2015-10-01

    Lactobacillus casei, a probiotic, and Streptococcus thermophilus, a fast acidifying lactic acid bacterial strain, are both used in the food industry. The aim of this study was to investigate the interaction between L. casei and S. thermophilus in the presence or absence of S. thermophilus-specific bacteriophage during milk fermentation. The acidification capability of L. casei co-cultured with S. thermophilus was significantly higher than that observed for L. casei or S. thermophilus cultured alone. However, the probiotic content (i.e., L. casei cell viability) was low. The fastest acidification and the highest viable L. casei cell count were observed in co-cultures of L. casei and S. thermophilus with S. thermophilus phage. In these co-cultures, S. thermophilus compensated for the slow acid production of L. casei in the early exponential growth phase. Thereafter, phage-induced lysis of the S. thermophilus cells eliminated the competition for nutrients, allowing L. casei to grow well. Additionally, the ruptured S. thermophilus cells released intracellular factors, which further promoted the growth and function of the probiotic bacteria. Crude cellular extract isolated from S. thermophilus also significantly accelerated the growth and propagation of L. casei, supporting the stimulatory role of the phage on this micro-ecosystem. PMID:26093989

  17. Lactobacillus casei Low-Temperature, Dairy-Associated Proteome Promotes Persistence in the Mammalian Digestive Tract.

    PubMed

    Lee, Bokyung; Tachon, Sybille; Eigenheer, Richard A; Phinney, Brett S; Marco, Maria L

    2015-08-01

    We found that incubation of probiotic Lactobacillus casei BL23 in milk at 4 °C prior to ingestion increased its survival in the mammalian digestive tract. To investigate the specific molecular adaptations of L. casei to milk, we used tandem mass spectrometry to compare proteins produced by L. casei BL23 at 4 °C in milk to those in exponential and stationary phase cells in laboratory culture medium at either 37 or 4 °C. These comparisons revealed a core of expressed L. casei proteins as well as proteins produced in either a growth-phase or temperature-specific manner. In total, 205 L. casei proteins were uniquely expressed or detected in higher abundance specifically as a result of incubation in milk and included an over-representation of proteins for cell surface modification, fatty acid metabolism, amino acid transport and metabolism, and inorganic ion transport. Genes for DltD (d-alanine transfer protein), FabH (3-oxoacyl-ACP synthase), RecA (recombinase A), and Sod (superoxide dismutase) were targeted for inactivation. The competitive fitness of the mutants was altered in the mouse intestine compared with wild-type cells. These results show that the food matrix can have a profound influence on dietary (probiotic) bacteria and their functional significance in the mammalian gut. PMID:26148687

  18. Improvement of exopolysaccharide production in Lactobacillus casei LC2W by overexpression of NADH oxidase gene.

    PubMed

    Li, Nan; Wang, Yuanlong; Zhu, Ping; Liu, Zhenmin; Guo, Benheng; Ren, Jing

    2015-02-01

    Lactobacillus casei LC2W is an exopolysaccharide (EPS)-producing strain with probiotic effects. To investigate the regulation mechanism of EPS biosynthesis and to improve EPS production through cofactor engineering, a H?O-forming NADH oxidase gene was cloned from Streptococcus mutans and overexpressed in L. casei LC2W under the control of constitutive promoter P??. The recombinant strain LC-nox exhibited 0.854 U/mL of NADH oxidase activity, which was elevated by almost 20-fold in comparison with that of wild-type strain. As a result, overexpression of NADH oxidase resulted in a reduction in growth rate. In addition, lactate production was decreased by 22% in recombinant strain. It was proposed that more carbon source was saved and used for the biosynthesis of EPS, the production of which was reached at 219.4 mg/L, increased by 46% compared to that of wild-type strain. This work provided a novel and convenient genetic approach to manipulate metabolic flux and to increase EPS production. To the best of our knowledge, this is the first report which correlates cofactor engineering with EPS production. PMID:25644955

  19. Safety and mutagenicity evaluation of Vigiis 101 powder made from Lactobacillus paracasei subsp. paracasei NTU 101.

    PubMed

    Tseng, Wei-Ting; Shih, Tsung-Wei; Liu, Shing-Hwa; Pan, Tzu-Ming

    2015-03-01

    The aim of the present work was to assess the genotoxic activity and the potential for toxicity upon repeated dosing of "Vigiis 101" powder, a probiotic consisting of dried bacteria Lactobacillus paracasei subsp. paracasei NTU 101. Results of the Ames test in Salmonella typhimurium strains TA1537, TA98, TA100, TA102, and TA1535 showed that Vigiis 101 (?5 mg per plate) was not mutagenic. We used experiments on ICR mice to evaluate the genotoxicity of Vigiis 101. Compared to the control, high-dose Vigiis 101 administration (16.72 g per kg of body weight) did not cause significant changes either in the number of reticulocytes or in the percentage (occurrence) of micronucleated reticulocytes. A mammalian chromosomal aberration test showed that the number of Chinese hamster ovary cells with abnormal chromosomes was <4% after Vigiis 101 treatment (maximal concentration was 5 mg/ml). A 28-day oral toxicity assay in Wistar rats was performed to assess the no-observed-adverse-effect level of Vigiis 101. Compared to the control, high-dose Vigiis 101 administration (5000 mg/kg/day) had no effects on mortality and body weight and did not cause toxicopathological lesions. Taken together, these results show that Vigiis 101 has no significant mutagenic or toxic effects. PMID:25481278

  20. Characterization of an exopolysaccharide produced by Lactobacillus plantarum YW11 isolated from Tibet Kefir.

    PubMed

    Wang, Ji; Zhao, Xiao; Tian, Zheng; Yang, Yawei; Yang, Zhennai

    2015-07-10

    An exopolysaccharide (EPS)-producing strain YW11 isolated from Tibet Kefir was identified as Lactobacillus plantarum, and the strain was shown to produce 90 mgL(-1) of EPS when grown in a semi-defined medium. The molecular mass of the EPS was 1.1 × 10(5)Da. The EPS was composed of glucose and galactose in a molar ratio of 2.71:1, with possible presence of N-acetylated sugar residues in the polysaccharide as confirmed by NMR spectroscopy. Rheological studies showed that the EPS had higher viscosity in skim milk, at lower temperature, or at acidic pH. The viscous nature of the EPS was confirmed by observation with scanning electron microscopy that demonstrated a highly branched and porous structure of the polysaccharide. The atomic force microscopy of the EPS further revealed presence of many spherical lumps, facilitating binding with water in aqueous solution. The EPS had a higher degradation temperature (287.7°C), suggesting high thermal stability of the EPS. PMID:25857955

  1. FTIR spectroscopy structural analysis of the interaction between Lactobacillus kefir S-layers and metal ions

    NASA Astrophysics Data System (ADS)

    Gerbino, E.; Mobili, P.; Tymczyszyn, E.; Fausto, R.; Gómez-Zavaglia, A.

    2011-02-01

    FTIR spectroscopy was used to structurally characterize the interaction of S-layer proteins extracted from two strains of Lactobacillus kefir (the aggregating CIDCA 8348 and the non-aggregating JCM 5818) with metal ions (Cd +2, Zn +2, Pb +2 and Ni +2). The infrared spectra indicate that the metal/protein interaction occurs mainly through the carboxylate groups of the side chains of Asp and Glut residues, with some contribution of the NH groups belonging to the peptide backbone. The frequency separation between the ?COO - anti-symmetric and symmetric stretching vibrations in the spectra of the S-layers in presence of the metal ions was found to be ca. 190 cm -1 for S-layer CIDCA 8348 and ca. 170 cm -1 for JCM 5818, denoting an unidentate coordination in both cases. Changes in the secondary structures of the S-layers induced by the interaction with the metal ions were also noticed: a general trend to increase the amount of ?-sheet structures and to reduce the amount of ?-helices was observed. These changes allow the proteins to adjust their structure to the presence of the metal ions at minimum energy expense, and accordingly, these adjustments were found to be more important for the bigger ions.

  2. Microencapsulation of Lactobacillus plantarum DKL 109 using External Ionic Gelation Method

    PubMed Central

    Chun, Honam

    2014-01-01

    The aim of this study was to apply the external ionic gelation using an atomizing spray device comprised of a spray gun to improve the viability of Lactobacillus plantarum DKL 109 and for its commercial use. Three coating material formulas were used to microencapsulate L. plantarum DKL 109: 2% alginate (Al), 1% alginate/1% gellan gum (Al-GG), and 1.5% alginate/3% gum arabic (Al-GA). Particle size of microcapsules was ranged from 18.2 to 23.01 ?m depending on the coating materials. Al-GA microcapsules showed the highest microencapsulation yield (98.11%) and resulted in a significant increase in survivability of probiotic in a high acid and bile environment. Encapsulation also improved the storage stability of cells. The viability of encapsulated cells remained constant after 1-mon storage at ambient temperature. The external ionic gelation method using an atomizing spray device and the Al-GA seems to be an efficient encapsulation technology for protecting probiotics in terms of scale-up potential and small microcapsule size.

  3. The potential of biodetoxification activity as a probiotic property of Lactobacillus reuteri.

    PubMed

    van Niel, Ed W J; Larsson, Christer U; Lohmeier-Vogel, Elke M; Rådström, Peter

    2012-01-16

    Previous work on the metabolism of Lactobacillus reuteri ATCC 55730 anticipated a variability in the use of organic electron acceptors as a means to relieve metabolic redox problems. Therefore, investigations focusing on this unique metabolism of L. reuteri may reveal a basis for new probiotic properties. For instance, L. reuteri may use reactive aldehydes and ketones as electron acceptors to balance their redox metabolism, which opens the possibility to exploit this bacterium for in vivo bioreduction of deleterious compounds in the gastrointestinal tract (GIT). Herein we demonstrate that L. reuteri ATCC 55730 cultures on glucose are able to use furfural (1g/L), and hydroxymethylfurfural (HMF) (0.5g/L), as electron acceptors. The former enhances the growth rate by about 25% and biomass yield by 15%, whereas the latter is inhibitory. Furfural is stoichiometrically reduced to furfuryl alcohol by the culture. The conversion of furfural had no effect on the flux distribution between the simultaneously operating phosphoketolase and Embden-Meyerhof pathways, but initiated a flux to acetate production. In addition to furfural and HMF, cellular extracts showed potential to reoxidize NADH and/or NADPH with acrolein, crotonaldehyde, and diacetyl, indicating that conversion reactions take place intracellularly, however, utilization mechanisms for the latter compounds may not be present in this strain. The strain did not reduce other GIT-related reactive compounds, including acrylamide, glyoxal, and furan. PMID:22071286

  4. Effect of Bile Salt Hydrolase Inhibitors on a Bile Salt Hydrolase from Lactobacillus acidophilus

    PubMed Central

    Lin, Jun; Negga, Rekek; Zeng, Ximin; Smith, Katie

    2014-01-01

    Bile salt hydrolase (BSH), a widely distributed function of the gut microbiota, has a profound impact on host lipid metabolism and energy harvest. Recent studies suggest that BSH inhibitors are promising alternatives to antibiotic growth promoters (AGP) for enhanced animal growth performance and food safety. Using a high-purity BSH from Lactobacillus salivarius strain, we have identified a panel of BSH inhibitors. However, it is still unknown if these inhibitors also effectively inhibit the function of the BSH enzymes from other bacterial species with different sequence and substrate spectrum. In this study, we performed bioinformatics analysis and determined the inhibitory effect of identified BSH inhibitors on a BSH from L. acidophilus. Although the L. acidophilus BSH is phylogenetically distant from the L. salivarius BSH, sequence analysis and structure modeling indicated the two BSH enzymes contain conserved, catalytically important amino residues and domain. His-tagged recombinant BSH from L. acidophilus was further purified and used to determine inhibitory effect of specific compounds. Previously identified BSH inhibitors also exhibited potent inhibitory effects on the L. acidophilus BSH. In conclusion, this study demonstrated that the BSH from L. salivarius is an ideal candidate for screening BSH inhibitors, the promising alternatives to AGP for enhanced feed efficiency, growth performance and profitability of food animals. PMID:25526498

  5. Controlled release of Lactobacillus rhamnosus biofilm probiotics from alginate-locust bean gum microcapsules.

    PubMed

    Cheow, Wean Sin; Kiew, Tie Yi; Hadinoto, Kunn

    2014-03-15

    Chitosan-coated alginate microcapsules containing high-density biofilm Lactobacillus rhamnosus have been previously shown to exhibit higher freeze drying- and thermal-tolerance than their planktonic counterparts. However, their cell release profile remains poor due to the capsules' susceptibility to the gastric environment. Herein the effects of adding locust bean (LB) and xanthan (XT) gums to alginate (AGN) capsules on the stress tolerance and cell release profiles in simulated gastrointestinal fluids are investigated. Compared to the AGN-only capsules, the AGN-LB capsules exhibit improved stress tolerance (i.e. ? 6x for freeze drying, 100x for thermotolerance, 10x for acid), whereas the AGN-XT capsules only improve the acid tolerance. Importantly, the AGN-LB capsules possess the optimal cell release profile with a majority of cells released in the simulated intestinal juice than in the gastric juice. The AGN-LB capsules' superiority is attributed to their stronger interaction with the chitosan coating and high swelling capacity, thus delaying their bulk dissolution. PMID:24528770

  6. Synbiotic yogurt-ice cream produced via incorporation of microencapsulated lactobacillus acidophilus (la-5) and fructooligosaccharide.

    PubMed

    Ahmadi, Abbas; Milani, Elnaz; Madadlou, Ashkan; Mortazavi, Seyed Ali; Mokarram, Reza Rezaei; Salarbashi, Davoud

    2014-08-01

    Yogurt-ice cream is a nutritious product with a refreshing taste and durability profoundly longer than that of yogurt. The probiotic Lactobacillus acidophilus (La-5) cells either in free or encapsulated form were incorporated into yog-ice cream and their survivability were studied. Fructooligosaccharide (FOS) as a prebiotic compound at three levels (0, 4 & 8 % w/w) was added to yogurt-ice cream mix and its effects on some chemical properties, overrun and firmness of product were evaluated. The higher the incorporated FOS concentration, the lower were the pH value and higher the total solid content of treatments. FOS incorporation (8 %) significantly increased the overrun of treatments and reduced their firmness. The viable counts of free probiotics decreased from ~9.55 to ~7.3 log cfu/g after 60 days of frozen storage while that of encapsulated cells merely decreased less than 1 log cycle. Encapsulation with alginate microbeads protected the probiotic cells against injuries in the freezing stage as well as, during frozen storage. PMID:25114349

  7. Effect of Nanometric Lactobacillus plantarum in Kimchi on Dextran Sulfate Sodium-Induced Colitis in Mice.

    PubMed

    Lee, Hyun Ah; Bong, Yeon-Ju; Kim, Hyunung; Jeong, Ji-Kang; Kim, Hee-Young; Lee, Kwang-Won; Park, Kun-Young

    2015-10-01

    Nanometric Lactobacillus plantarum (nLp) is a processed form of Lab. plantarum derived from kimchi and is 0.5-1.0??m in size. This study was undertaken to determine the effect of nLp and kimchi plus nLp (K-nLp) on a dextran sulfate sodium (DSS)-induced mouse model of colitis. Animals fed nLp or K-nLp had longer colons, but lower colon weights per unit length than DSS controls. In addition, nLp- or K-nLp-fed animals showed lower levels of proinflammatory cytokines and inflammatory genes in serum and in colon tissues, lower populations of total bacteria, but higher populations of lactic acid bacteria in feces, and lower activities of fecal ?-glucosidase and ?-glucuronidase. Furthermore, these suppressive activities of nLp on colitis were equivalent to or higher than those of naive Lab. plantarum. Consequently, nLp was found to exhibit anticolitic effects, and the addition of nLp to kimchi was found to enhance the protective activity of kimchi against DSS-induced colitis. These results suggest that nLp might be an effective substitute for live probiotics and be useful as a functional ingredient with the anticolitic activity by the probiotic and food processing industries. PMID:26305853

  8. Characterization and bioactivities of an exopolysaccharide produced by Lactobacillus plantarum YW32.

    PubMed

    Wang, Ji; Zhao, Xiao; Yang, Yawei; Zhao, Aimei; Yang, Zhennai

    2015-03-01

    The exopolysaccharide (EPS) produced by Lactobacillus plantarum YW32 was purified and characterized, and the in vitro bioactivities of the purified EPS were also evaluated. The EPS had a molecular weight of 1.03×10(5) Da, and it consisted of mannose, fructose, galactose and glucose in an approximate molar ratio of 8.2:1:4.1:4.2. Microstructural studies of the EPS demonstrated a web-like structure composed of compact ropes, and presence of many homogeneous rod-shaped lumps. The EPS also showed high thermal stability with a degradation temperature of 283.5°C. Furthermore, the EPS at a dose of 5mg/ml had strong scavenging abilities toward hydroxyl (77.5%) and superoxide radicals (66.5%). The EPS exhibited a concentration-dependent inhibitory effect on the formation of biofilms by several pathogenic bacteria, including Escherichia coli O157, Shigella flexneri CMCC (B), Staphylococcus aureus AC1 and Salmonella typhimurium S50333. In vitro antitumor assay of the EPS showed that it had good inhibitory activity against colon cancer HT-29 cells. These characteristics and bioactivities of the EPS would make it a promising candidate for use as a potential food adjunct in foods with healthy properties. PMID:25532782

  9. Lactobacillus rhamnosus HN001 decreases the severity of necrotizing enterocolitis in neonatal mice and preterm piglets: evidence in mice for a role of TLR9

    PubMed Central

    Good, Misty; Sodhi, Chhinder P.; Ozolek, John A.; Buck, Rachael H.; Goehring, Karen C.; Thomas, Debra L.; Vikram, Amit; Bibby, Kyle; Morowitz, Michael J.; Firek, Brian; Lu, Peng

    2014-01-01

    Necrotizing enterocolitis (NEC) is the leading cause of death from gastrointestinal disease in premature infants and develops partly from an exaggerated intestinal epithelial immune response to indigenous microbes. There has been interest in administering probiotic bacteria to reduce NEC severity, yet concerns exist regarding infection risk. Mechanisms of probiotic activity in NEC are unknown although activation of the microbial DNA receptor Toll-like receptor-9 (TLR9) has been postulated. We now hypothesize that the Gram-positive bacterium Lactobacillus rhamnosus HN001 can attenuate NEC in small and large animal models, that its microbial DNA is sufficient for its protective effects, and that protection requires activation of the Toll-like receptor 9 (TLR9). We now show that oral administration of live or UV-inactivated Lactobacillus rhamnosus HN001 attenuates NEC severity in newborn mice and premature piglets, as manifest by reduced histology score, attenuation of mucosal cytokine response, and improved gross morphology. TLR9 was required for Lactobacillus rhamnosus-mediated protection against NEC in mice, as the selective decrease of TLR9 from the intestinal epithelium reversed its protective effects. Strikingly, DNA of Lactobacillus rhamnosus HN001 reduced the extent of proinflammatory signaling in cultured enterocytes and in samples of resected human ileum ex vivo, suggesting the therapeutic potential of this probiotic in clinical NEC. Taken together, these findings illustrate that Lactobacillus rhamnosus HN001 is an effective probiotic for NEC via activation of the innate immune receptor TLR9 and that Lactobacillus rhamnosus DNA is sufficient for its protective effects, potentially reducing concerns regarding the infectious risk of this novel therapeutic approach. PMID:24742987

  10. Effect of aqueous and alcoholic Stevia (Stevia rebaudiana) extracts against Streptococcus mutans and Lactobacillus acidophilus in comparison to chlorhexidine: An in vitro study

    PubMed Central

    Ajagannanavar, Sunil Lingaraj; Shamarao, Supreetha; Battur, Hemant; Tikare, Shreyas; Al-Kheraif, Abdulaziz Abdullah; Al Sayed, Mohammed Sayed Al Esawy

    2014-01-01

    Introduction: Stevia (S. rebaudiana) a herb which has medicinal value and was used in ancient times as a remedy for a great diversity of ailments and sweetener. Leaves of Stevia contain a high concentration of Stevioside and Rebaudioside which are supposed to be sweetening agents. Aim: To compare the efficacy of aqueous and alcoholic S. rebaudiana extract against Streptococcus mutans and Lactobacillus acidophilus in comparison to chlorhexidine. Materials and Methods: In the first part of the study, various concentrations of aqueous and ethanolic Stevia extract were prepared in the laboratory of Pharmacy College. It was then subjected to microbiological assay to determine its zone of inhibition using Agar disk diffusion test and minimum inhibitory concentration (MIC) using serial broth dilution method against Streptococcus mutans and Lactobacillus acidophilus. Chlorhexidine was used as a positive control. One way Analysis of Variance (ANOVA) test was used for multiple group comparisons followed by Tukey post hoc for group wise comparisons. Results: Minimum inhibitory concentration (MIC) of aqueous and ethnolic Stevia extract against Streptococcus mutans and Lactobacillus acidophilus were 25% and 12.5% respectively. Mean zone of inhibition of the aqueous and alcoholic Stevia extracts against Streptococcus mutans at 48 hours were 22.8 mm and 26.7 mm respectively. Mean zone of inhibition of the aqueous and alcoholic Stevia extracts against Lactobacillus acidophilus at 48 hours were 14.4 mm and 15.1 mm respectively. Mean zone of inhibition of the chlorhexidine against Streptococcus mutans and Lactobacillus acidophilus at 48 hours was 20.5 and 13.2 respectively. Conclusion: The inhibitory effect shown by alcoholic Stevia extract against Streptococcus mutans and Lactobacillus acidophilus was superior when compared with that of aqueous form and was inferior when compared with Chlorhexidine. PMID:25558451

  11. Lactobacillus rhamnosus HN001 decreases the severity of necrotizing enterocolitis in neonatal mice and preterm piglets: evidence in mice for a role of TLR9.

    PubMed

    Good, Misty; Sodhi, Chhinder P; Ozolek, John A; Buck, Rachael H; Goehring, Karen C; Thomas, Debra L; Vikram, Amit; Bibby, Kyle; Morowitz, Michael J; Firek, Brian; Lu, Peng; Hackam, David J

    2014-06-01

    Necrotizing enterocolitis (NEC) is the leading cause of death from gastrointestinal disease in premature infants and develops partly from an exaggerated intestinal epithelial immune response to indigenous microbes. There has been interest in administering probiotic bacteria to reduce NEC severity, yet concerns exist regarding infection risk. Mechanisms of probiotic activity in NEC are unknown although activation of the microbial DNA receptor Toll-like receptor-9 (TLR9) has been postulated. We now hypothesize that the Gram-positive bacterium Lactobacillus rhamnosus HN001 can attenuate NEC in small and large animal models, that its microbial DNA is sufficient for its protective effects, and that protection requires activation of the Toll-like receptor 9 (TLR9). We now show that oral administration of live or UV-inactivated Lactobacillus rhamnosus HN001 attenuates NEC severity in newborn mice and premature piglets, as manifest by reduced histology score, attenuation of mucosal cytokine response, and improved gross morphology. TLR9 was required for Lactobacillus rhamnosus-mediated protection against NEC in mice, as the selective decrease of TLR9 from the intestinal epithelium reversed its protective effects. Strikingly, DNA of Lactobacillus rhamnosus HN001 reduced the extent of proinflammatory signaling in cultured enterocytes and in samples of resected human ileum ex vivo, suggesting the therapeutic potential of this probiotic in clinical NEC. Taken together, these findings illustrate that Lactobacillus rhamnosus HN001 is an effective probiotic for NEC via activation of the innate immune receptor TLR9 and that Lactobacillus rhamnosus DNA is sufficient for its protective effects, potentially reducing concerns regarding the infectious risk of this novel therapeutic approach. PMID:24742987

  12. Taxonomic Structure and Monitoring of the Dominant Population of Lactic Acid Bacteria during Wheat Flour Sourdough Type I Propagation Using Lactobacillus sanfranciscensis Starters?

    PubMed Central

    Siragusa, Sonya; Di Cagno, Raffaella; Ercolini, Danilo; Minervini, Fabio; Gobbetti, Marco; De Angelis, Maria

    2009-01-01

    The structure and stability of the dominant lactic acid bacterium population were assessed during wheat flour sourdough type I propagation by using singly nine strains of Lactobacillus sanfranciscensis. Under back-slopping propagation with wheat flour type 0 F114, cell numbers of presumptive lactic acid bacteria varied slightly between and within starters. As determined by randomly amplified polymorphic DNA-PCR and restriction endonuclease analysis-pulsed-field gel electrophoresis analyses, only three (LS8, LS14, and LS44) starters dominated throughout 10 days of propagation. The others progressively decreased to less than 3 log CFU g?1. Partial sequence analysis of the 16S rRNA and recA genes and PCR-denaturating gradient gel electrophoresis analysis using the rpoB gene allowed identification of Weissella confusa, Lactobacillus sanfranciscensis, Lactobacillus plantarum, Lactobacillus rossiae, Lactobacillus brevis, Lactococcus lactis subsp. lactis, Pediococcus pentosaceus, and Lactobacillus spp. as the dominant species of the raw wheat flour. At the end of propagation, one autochthonous strain of L. sanfranciscensis was found in all the sourdoughs. Except for L. brevis, strains of the above species were variously found in the mature sourdoughs. Persistent starters were found in association with other biotypes of L. sanfranciscensis and with W. confusa or L. plantarum. Sourdoughs were characterized for acidification, quotient of fermentation, free amino acids, and community-level catabolic profiles by USING Biolog 96-well Eco microplates. In particular, catabolic profiles of sourdoughs containing persistent starters behaved similarly and were clearly differentiated from the others. The three persistent starters were further used for the production of sourdoughs and propagated by using another wheat flour whose lactic acid bacterium population in part differed from the previous one. Also, in this case all three starter strains persisted during propagation. PMID:19088320

  13. Identification of lactic acid bacteria involved in the spoilage of pasteurized "foie gras" products.

    PubMed

    Matamoros, S; André, S; Hue, I; Prévost, H; Pilet, M F

    2010-07-01

    The spoiling microflora of a re-packaged French "foie gras" product was studied. A total of 54 isolates, originating from two different factories, were identified using phenotypical and molecular methods (partial 16S rDNA sequencing). Weissella viridescens was the main species detected in the products from factory 1 (64% of the isolates). These products had a low lactic acid concentration and were considered as non-spoiled. The microflora of factory 2 was dominated mainly by the genus Lactobacillus (95% of the isolates), and the high lactic acid concentration of these products was linked with a strong spoilage. Among the 30 Lactobacillus strains, three species were predominant: Lactobacillus sakei (nine isolates), Lactobacillus coryniformis (eight isolates) and Lactobacillus paraplantarum (five isolates). Challenge tests were performed to confirm the involvement of the Lactobacillus strains in the spoilage of the product. Sterile "foie gras" samples were inoculated with 14 LAB strains from the collection. The most acidifying strains belonged to the species L. sakei, Lactobacillus plantarum and L. paraplantarum. This confirmed the role of the strains from the Lactobacillus genus as the main spoilers of "foie gras" products and will be useful to design new quality protocols and extend the shelf-life of these products. PMID:20416816

  14. Carbohydrate catabolic flexibility in the mammalian intestinal commensal Lactobacillus ruminis revealed by fermentation studies aligned to genome annotations

    PubMed Central

    2011-01-01

    Background Lactobacillus ruminis is a poorly characterized member of the Lactobacillus salivarius clade that is part of the intestinal microbiota of pigs, humans and other mammals. Its variable abundance in human and animals may be linked to historical changes over time and geographical differences in dietary intake of complex carbohydrates. Results In this study, we investigated the ability of nine L. ruminis strains of human and bovine origin to utilize fifty carbohydrates including simple sugars, oligosaccharides, and prebiotic polysaccharides. The growth patterns were compared with metabolic pathways predicted by annotation of a high quality draft genome sequence of ATCC 25644 (human isolate) and the complete genome of ATCC 27782 (bovine isolate). All of the strains tested utilized prebiotics including fructooligosaccharides (FOS), soybean-oligosaccharides (SOS) and 1,3:1,4-?-D-gluco-oligosaccharides to varying degrees. Six strains isolated from humans utilized FOS-enriched inulin, as well as FOS. In contrast, three strains isolated from cows grew poorly in FOS-supplemented medium. In general, carbohydrate utilisation patterns were strain-dependent and also varied depending on the degree of polymerisation or complexity of structure. Six putative operons were identified in the genome of the human isolate ATCC 25644 for the transport and utilisation of the prebiotics FOS, galacto-oligosaccharides (GOS), SOS, and 1,3:1,4-?-D-Gluco-oligosaccharides. One of these comprised a novel FOS utilisation operon with predicted capacity to degrade chicory-derived FOS. However, only three of these operons were identified in the ATCC 27782 genome that might account for the utilisation of only SOS and 1,3:1,4-?-D-Gluco-oligosaccharides. Conclusions This study has provided definitive genome-based evidence to support the fermentation patterns of nine strains of Lactobacillus ruminis, and has linked it to gene distribution patterns in strains from different sources. Furthermore, the study has identified prebiotic carbohydrates with the potential to promote L. ruminis growth in vivo. PMID:21995520

  15. Evaluation of the functional potential of Weissella and Lactobacillus isolates obtained from Nigerian traditional fermented foods and cow's intestine.

    PubMed

    Ayeni, Funmilola A; Sánchez, Borja; Adeniyi, Bolanle A; de Los Reyes-Gavilán, Clara G; Margolles, Abelardo; Ruas-Madiedo, Patricia

    2011-05-27

    The characterisation of 24 lactic acid bacteria (LAB) isolates from Nigerian traditional fermented dairy foods, including some cow's intestine isolates, was conducted in order to select isolates for potential use as probiotics. LAB isolates were identified by partial sequencing the 16S rRNA gene as belonging to the species Lactobacillus paracasei, Lactobacillus brevis and mainly Weissella confusa. At the end of a characterisation process, 2 L. paracasei and 2 W. confusa isolates were selected, and their resistance to a simulated gastrointestinal digestion and their ability to adhere to eukaryotic cell lines were assessed. The survival to the simulated gastrointestinal passage was higher when bacterial suspensions were made in skimmed milk (2.0±0.8 log units reduction) or at the simulated gastric juice pH 3 (2.7±0.9 log units reduction) than at pH 2.0 (5.5±0.7 log units reduction). Adhesion of LAB to both intestinal and vaginal epithelial models was comparable or higher than that of the reference Lactobacillus rhamnosus GG. However, some of the isolates increased the adhesion of the pathogen Escherichia coli LMG2092 to HT-29 and HeLa monolayers. Overall, isolates L. paracasei UI14 and W. confusa UI7 are good candidates for further studying potential benefits that support their use as probiotics. This is one of the few articles reporting the characterisation and the probiotic potential of Weissella, although more studies are needed in order to establish their safety for potential probiotic applications. PMID:21482440

  16. In vitro screening of selected probiotic properties of Lactobacillus strains isolated from traditional fermented cabbage and cucumber.

    PubMed

    Zieli?ska, Dorota; Rzepkowska, Anna; Radawska, Anna; Zieli?ski, Konrad

    2015-02-01

    Most important during probiotic selection are gastric acid and bile tolerance, the adhesion to the luminal epithelium to colonize the lower gastrointestinal tract of a human and safety for human consumption. The aim of this study was to evaluate the selected probiotic in vitro properties of Lactobacillus spp. Strains isolated from traditional fermented food. A total 38 strains were isolated from the pickled samples and 14 were identified as Lactobacillus spp. The survival of almost all strains after incubation at pH 2.5 did not change markedly, and remained at above 90 % (10(9) CFU/mL). The strains also exhibited a high survival rate at pH 3.5 (>90 %), whereas pH 1.5 all were died. Just four strains could survive 90 min. at pH 1.5 (<39 %). The incubation with 0.2 % bile salt solution resulted in a survival rates of 81-94 % after 24 h, whereas after incubation in 2 and 4 % bile salt solution it was 59-94 %. All tested strains showed very good and good resistance to 0.4 % phenol addition, however only Lb. johnsonii K4 was able to multiply. The hydrophobic nature of the cell surface of the tested strains was moderated recording hydrophobicity of Lb. johnsonii K4 and Lb. rhamnosus K3 above 60 %. Safety evaluation excluded four of tested strains as candidate probiotics, according to antibiotic resistance patterns and certain metabolic activities. On the basis on the results 10 of the selected Lactobacillus strains are safe and can survive under gastrointestinal conditions, which requires them to future in vitro and in vivo probiotic studies. PMID:25270682

  17. Analysis of the Fecal Microflora of Human Subjects Consuming a Probiotic Product Containing Lactobacillus rhamnosus DR20

    PubMed Central

    Tannock, G. W.; Munro, K.; Harmsen, H. J. M.; Welling, G. W.; Smart, J.; Gopal, P. K.

    2000-01-01

    The composition of the fecal microflora of 10 healthy subjects was monitored before (6-month control period), during (6-month test period), and after (3-month posttest period) the administration of a milk product containing Lactobacillus rhamnosus DR20 (daily dose, 1.6 × 109 lactobacilli). Monthly fecal samples were examined by a variety of methods, including bacteriological culture analysis, fluorescent in situ hybridization with group-specific DNA probes, denaturing gradient gel electrophoresis of the V2-V3 region of 16S rRNA genes amplified by PCR, gas-liquid chromatography, and bacterial enzyme activity analysis. The composition of the Lactobacillus population of each subject was analyzed by pulsed-field gel electrophoresis of bacterial DNA digests in order to differentiate between DR20 and other strains present in the samples. Representative isolates of lactobacilli were identified to the species level by sequencing the V2-V3 region of their 16S rRNA genes and comparing the sequences obtained (BLAST search) to sequences in the GenBank database. DR20 was detected in the feces of all of the subjects during the test period, but at different frequencies. The presence of DR20 among the numerically predominant strains was related to the presence or absence of a stable indigenous population of lactobacilli during the control period. Strain DR20 did not persist at levels of >102 cells per g in the feces of most of the subjects after consumption of the product ceased; the only exception was one subject in which this strain was detected for 2 months during the posttest period. We concluded that consumption of the DR20-containing milk product transiently altered the Lactobacillus and enterococcal contents of the feces of the majority of consumers without markedly affecting biochemical or other bacteriological factors. PMID:10831441

  18. Evaluation of Synergistic Interactions Between Cell-Free Supernatant of Lactobacillus Strains and Amikacin and Genetamicin Against Pseudomonas aeruginosa

    PubMed Central

    Aminnezhad, Sargol; Kermanshahi, Rouha Kasra; Ranjbar, Reza

    2015-01-01

    Background: The indiscriminate use of antibiotics in the treatment of infectious diseases can increase the development of antibiotic resistance. Therefore, there is a big demand for new sources of antimicrobial agents and alternative treatments for reduction of antibiotic dosage required to decrease the associated side effects. Objectives: In this study, the synergistic action of aminoglycoside antibiotics and cell-free supernatant (CFS) of probiotic (Lactobacillus rahmnosus and L. casei) against Pseudomonas aeruginosa PTCC 1430 was evaluated. Materials and Methods: A growth medium for culturing of probiotic bacteria was separated by centrifugation. The antimicrobial effects of CFS of probiotic bacteria were evaluated using the agar well diffusion assay. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were evaluated using the micro dilution method. Finally, an interaction between CFS and amikacin or gentamicin against P. aeruginosa PTCC 1430 was examined through the checkerboard method and fractional inhibitory concentration (FIC). Furthermore, CFSs from Lactobacillus strains were analyzed by reversed phase HPLC (RP-HPLC) for antimicrobial compounds. Results: The results showed a significant effect of CFS on the growth of P. aeruginosa. The MIC and MBC of CFS from L. casei were 62.5 µL?mL while the MIC and MBC of CFS from L. rhamnosus were 62.5 ?L?mL and 125 ?L?mL, respectively. Using the FIC indices, synergistic interactions were observed in combination of CFS and antibiotics. Fractional Inhibitory Concentration indices of CFS from L. casei and aminoglycoside antibiotics were 0.124 and 0.312 while FIC indices of CFS from L. rhamnosus and aminoglycoside antibiotics were 0.124 and 0.56, respectively showing a synergism effect. The results of RP-HPLC showed that CFS of Lactobacillus strains contained acetic acid, lactic acid and hydrogen peroxide (H2O2). Conclusions: Our findings indicate that probiotic bacterial strains of Lactobacillus have a significant inhibitory effect on the growth of P. aeruginosa PTCC 1430. The antimicrobial potency of this combination can be useful for designing and developing alternative therapeutic strategies against P. aeruginosa infections. PMID:26034539

  19. Rapid detection of Lactobacillus kefiranofaciens in kefir grain and kefir milk using newly developed real-time PCR.

    PubMed

    Kim, Dong-Hyeon; Chon, Jung-Whan; Kim, Hong-Seok; Yim, Jin-Hyeok; Kim, Hyunsook; Seo, Kun-Ho

    2015-04-01

    Lactobacillus kefiranofaciens is an indicator microorganism for kefir and a key factor in kefir grain formation and kefiran production. We designed a novel real-time PCR primer and probe set, LKF_KU504, for the rapid detection of L. kefiranofaciens. In inclusivity and exclusivity tests, only 14 L. kefiranofaciens strains were positive among 61 microorganisms, indicating 100 % sensitivity and specificity. The LKF_KU504 set also differentiated kefir milk from 30 commercial nonkefir yogurts. The levels of L. kefiranofaciens in kefir grain and kefir milk were significantly different, indicating L. kefiranofaciens was more concentrated in kefir grain than in kefir milk. PMID:25836417

  20. Effect of Lactobacillus salivarius Bacteriocin Abp118 on the Mouse and Pig Intestinal Microbiota

    PubMed Central

    Riboulet-Bisson, Eliette; Sturme, Mark H. J.; Jeffery, Ian B.; O'Donnell, Michelle M.; Neville, B. Anne; Forde, Brian M.; Claesson, Marcus J.; Harris, Hugh; Gardiner, Gillian E.; Casey, Patrick G.; Lawlor, Peadar G.; O'Toole, Paul W.; Ross, R. Paul

    2012-01-01

    Lactobacilli are Gram-positive bacteria that are a subdominant element in the human gastrointestinal microbiota, and which are commonly used in the food industry. Some lactobacilli are considered probiotic, and have been associated with health benefits. However, there is very little culture-independent information on how consumed probiotic microorganisms might affect the entire intestinal microbiota. We therefore studied the impact of the administration of Lactobacillus salivarius UCC118, a microorganism well characterized for its probiotic properties, on the composition of the intestinal microbiota in two model animals. UCC118 has anti-infective activity due to production of the bacteriocin Abp118, a broad-spectrum class IIb bacteriocin, which we hypothesized could impact the microbiota. Mice and pigs were administered wild-type (WT) L. salivarius UCC118 cells, or a mutant lacking bacteriocin production. The microbiota composition was determined by pyrosequencing of 16S rRNA gene amplicons from faeces. The data show that L. salivarius UCC118 administration had no significant effect on proportions of major phyla comprising the mouse microbiota, whether the strain was producing bacteriocin or not. However, L. salivarius UCC118 WT administration led to a significant decrease in Spirochaetes levels, the third major phylum in the untreated pig microbiota. In both pigs and mice, L. salivarius UCC118 administration had an effect on Firmicutes genus members. This effect was not observed when the mutant strain was administered, and was thus associated with bacteriocin production. Surprisingly, in both models, L. salivarius UCC118 administration and production of Abp118 had an effect on Gram-negative microorganisms, even though Abp118 is normally not active in vitro against this group of microorganisms. Thus L. salivarius UCC118 administration has a significant but subtle impact on mouse and pig microbiota, by a mechanism that seems at least partially bacteriocin-dependent. PMID:22363561

  1. Purification and characterization of a novel plantaricin, KL-1Y, from Lactobacillus plantarum KL-1.

    PubMed

    Rumjuankiat, Kittaporn; Perez, Rodney Horanda; Pilasombut, Komkhae; Keawsompong, Suttipun; Zendo, Takeshi; Sonomoto, Kenji; Nitisinprasert, Sunee

    2015-06-01

    Three bacteriocins from Lactobacillus plantarum KL-1 were successfully purified using ammonium sulfate precipitation, cation-exchange chromatography and reverse-phase HPLC. The bacteriocin peptides KL-1X, -1Y and -1Z had molecular masses of 3053.82, 3498.16 and 3533.16 Da, respectively. All three peptides were stable at pH 2-12 and 25 °C and at high temperatures of 80 and 100 °C for 30 min and 121 °C for 15 min. However, they differed in their susceptibility to proteolytic enzymes and their inhibition spectra. KL-1Y showed broad inhibitory activities against Gram-positive and Gram-negative bacteria, including Salmonella enterica serovar Enteritidis DMST 17368, Pseudomonas aeruginosa ATCC 15442, P. aeruginosa ATCC 9027, Escherichia coli O157:H7 and E. coli ATCC 8739. KL-1X and -1Z inhibited only Gram-positive bacteria. KL-1X, KL-1Y and KL-1Z exhibited synergistic activity. The successful amino acid sequencing of KL-1Y had a hydrophobicity of approximately 30 % and no cysteine residues suggested its novelty, and it was designated "plantaricin KL-1Y". Plantaricin KL-1Y exhibited bactericidal activity against Bacillus cereus JCM 2152(T). Compared to nisin, KL-1Y displayed broad inhibitory activities of 200, 800, 1600, 800, 400 and 400 AU/mL against the growth of Bacillus coagulans JCM 2257(T), B. cereus JCM 2152(T), Listeria innocua ATCC 33090(T), Staphylococcus aureus TISTR 118, E. coli O157:H7 and E. coli ATCC 8739, respectively, whereas nisin had similar activities against only B. coagulans JCM 2257(T) and B. cereus JCM 2152(T). Therefore, the novel plantaricin KL-1Y is a promising antimicrobial substance for food safety uses in the future. PMID:25862353

  2. Transcriptome signatures of class I and III stress response deregulation in Lactobacillus plantarum reveal pleiotropic adaptation

    PubMed Central

    2013-01-01

    Background To cope with environmental challenges bacteria possess sophisticated defense mechanisms that involve stress-induced adaptive responses. The canonical stress regulators CtsR and HrcA play a central role in the adaptations to a plethora of stresses in a variety of organisms. Here, we determined the CtsR and HrcA regulons of the lactic acid bacterium Lactobacillus plantarum WCFS1 grown under reference (28°C) and elevated (40°C) temperatures, using ctsR, hrcA, and ctsR-hrcA deletion mutants. Results While the maximum specific growth rates of the mutants and the parental strain were similar at both temperatures (0.33?±?0.02 h-1 and 0.34?±?0.03 h-1, respectively), DNA microarray analyses revealed that the CtsR or HrcA deficient strains displayed altered transcription patterns of genes encoding functions involved in transport and binding of sugars and other compounds, primary metabolism, transcription regulation, capsular polysaccharide biosynthesis, as well as fatty acid metabolism. These transcriptional signatures enabled the refinement of the gene repertoire that is directly or indirectly controlled by CtsR and HrcA of L. plantarum. Deletion of both regulators, elicited transcriptional changes of a large variety of additional genes in a temperature-dependent manner, including genes encoding functions involved in cell-envelope remodeling. Moreover, phenotypic assays revealed that both transcription regulators contribute to regulation of resistance to hydrogen peroxide stress. The integration of these results allowed the reconstruction of CtsR and HrcA regulatory networks in L. plantarum, highlighting the significant intertwinement of class I and III stress regulons. Conclusions Taken together, our results enabled the refinement of the CtsR and HrcA regulatory networks in L. plantarum, illustrating the complex nature of adaptive stress responses in this bacterium. PMID:24238744

  3. Lactobacillus casei Shirota enhances the preventive efficacy of soymilk in chemically induced breast cancer.

    PubMed

    Kaga, Chiaki; Takagi, Akimitsu; Kano, Mitsuyoshi; Kado, Shoichi; Kato, Ikuo; Sakai, Masashi; Miyazaki, Kouji; Nanno, Masanobu; Ishikawa, Fumiyasu; Ohashi, Yasuo; Toi, Masakazu

    2013-11-01

    Soy foods are known to be effective for breast cancer prevention. The habitual consumption of soy isoflavones in combination with the probiotic Lactobacillus casei Shirota (LcS) was shown to decrease the risk of breast cancer occurrence in our previous population-based case-controlled study among Japanese women. The present study aimed to elucidate the cooperative prevention mechanism of soymilk and LcS using an animal carcinogenic model. Female Sprague-Dawley rats received a high-fat, AIN-76A diet containing soymilk, LcS, both soymilk and LcS, or none and were orally exposed to 2-amino-1-methyl-6-penylimidazo[4,5-b]pyridine at a dose of 85 mg/kg bodyweight eight times for 2 weeks. The development of palpable mammary tumors was monitored for 17 weeks. Tumor tissues were immunohistochemically examined for estrogen receptor (ER)-?, Ki-67 and CD34. Compared with the control group, the incidence and multiplicity of mammary tumors were reduced by soymilk alone and soymilk in combination with LcS, while tumor volume was decreased by LcS alone and LcS in combination with soymilk. An immunohistochemical analysis revealed that soymilk in combination with LcS more effectively reduced the numbers of ER-?-positive and Ki-67-positive cells in tumors than soymilk alone and that both soymilk and LcS inhibited tumor angiogenesis. These results demonstrated that soymilk prevents the development of mammary tumors and that LcS suppresses tumor growth, potentially enhancing the preventive efficacy of soymilk. The habitual consumption of LcS in combination with soymilk might be a beneficial dietary style for breast cancer prevention. PMID:23992486

  4. Fortification of table olive packing with the potential probiotic bacteria Lactobacillus pentosus TOMC-LAB2

    PubMed Central

    Rodríguez-Gómez, F.; Romero-Gil, V.; García-García, P.; Garrido-Fernández, A.; Arroyo-López, Francisco N.

    2014-01-01

    Dairy products are currently the main carriers of probiotic microorganisms to the human body. However, the development of new matrices for probiotic delivery is convenient for intolerant to milk (or its derivatives) and those requiring low-cholesterol diet consumers. The present work focused on the fortification of previously fermented green Spanish style olives with the autochthonous putative probiotic bacteria Lactobacillus pentosus TOMC-LAB2. The fortification was carried out by inoculating the bacteria into the packing brines using Manzanilla fruits from three different processes: (i) spontaneously fermented (F1), (ii) fermented using L. pentosus TOMC-LAB2 as starter (F2), and (iii) spontaneously fermented and then thermally treated (F3). Data showed that all inoculated treatments had higher population levels (5.49, 4.41, and 6.77 log10 cfu/cm2) than their respective controls (1.66, 4.33, and 0.0 log10 cfu/cm2, for F1, F2, and F3 treatments, respectively). The presence of L. pentosus TOMC-LAB2 on olive surface was confirmed by rep-PCR, with a recovery frequency at the end of the shelf life (200 days) of 52.6, 57.9, and 100.0% for F1, F2, and F3 treatments, respectively. Thus, results obtained in this work show the ability of this microorganism to survive under packing conditions for long period of times as well as to colonize the olive surface which is the food finally ingested by consumers. This opens the possibility for the development of a new and simply probiotic fortified olive product. PMID:25232354

  5. Cystathionine ?-Lyase Is a Component of Cystine-Mediated Oxidative Defense in Lactobacillus reuteri BR11?

    PubMed Central

    Lo, Raquel; Turner, Mark S.; Barry, Daniel G.; Sreekumar, Revathy; Walsh, Terence P.; Giffard, Philip M.

    2009-01-01

    Lactobacillus reuteri BR11 possesses a novel mechanism of oxidative defense involving an abundant cystine ABC transporter encoded by the cyuABC gene cluster. Large amounts of thiols, including H2S, are secreted upon cystine uptake by the CyuC transporter. A cystathionine ?-lyase (cgl) gene is cotranscribed with the cyu genes in several L. reuteri strains and was hypothesized to participate in cystine-mediated oxidative defense by producing reducing equivalents. This hypothesis was tested with L. reuteri BR11 by constructing a cgl mutant (PNG901) and comparing it to a similarly constructed cyuC mutant (PNG902). Although Cgl was required for H2S production from cystine, it was not crucial for oxidative defense in de Mann-Rogosa-Sharpe medium, in contrast to CyuC, whose inactivation resulted in lag-phase arrest in aerated cultures. The importance of Cgl in oxidative defense was seen only in the presence of hemin, which poses severe oxidative stress. The growth defects in aerated cultures of both mutants were alleviated by supplementation with cysteine (and cystine in the cgl mutant) but not methionine, with the cyuC mutant showing a much higher concentration requirement. We conclude that L. reuteri BR11 requires a high concentration of exogenous cysteine/cystine to grow optimally under aerobic conditions. This requirement is fulfilled by the abundant CyuC transporter, which has probably arisen due to the broad substrate specificity of Cgl, resulting in a futile pathway which degrades cystine taken up by the CyuC transporter to H2S. Cgl plays a secondary role in oxidative defense by its well-documented function of cysteine biosynthesis. PMID:19124577

  6. Effects of Lactobacillus plantarum on gut barrier function in experimental obstructive jaundice

    PubMed Central

    Zhou, Yu-Kun; Qin, Huan-Long; Zhang, Ming; Shen, Tong-Yi; Chen, Hong-Qi; Ma, Yan-Lei; Chu, Zhao-Xin; Zhang, Peng; Liu, Zhi-Hua

    2012-01-01

    AIM: To investigate the mechanisms of Lactobacillus plantarum (L. plantarum) action on gut barrier in preoperative and postoperative experimental obstructive jaundice in rats. METHODS: Forty rats were randomly divided into groups of sham-operation, bile duct ligation (BDL), BDL + L. plantarum, BDL + internal biliary drainage (IBD), and BDL + IBD + L. plantarum. Ten days after L. plantarum administration, blood and ileal samples were collected from the rats for morphological examination, and intestinal barrier function, liver function, intestinal oxidative stress and protein kinase C (PKC) activity measurement. The distribution and expression of the PKC and tight junction (TJ) proteins, such as occludin, zonula occludens-1, claudin-1, claudin-4, junction adhesion molecule-A and F-actin, were examined by confocal laser scanning microscopy, immunohistochemistry, Western blotting, real-time fluorescent quantitative polymerase chain reaction assay. RESULTS: L. plantarum administration substantially restored gut barrier, decreased enterocyte apoptosis, improved intestinal oxidative stress, promoted the activity and expression of protein kinase (BDL vs BDL + L. plantarum, 0.295 ± 0.007 vs 0.349 ± 0.003, P < 0.05; BDL + IBD vs BDL + IBD + L. plantarum, 0.407 ± 0.046 vs 0.465 ± 0.135, P < 0.05), and particularly enhanced the expression and phosphorylation of TJ proteins in the experimental obstructive jaundice (BDL vs BDL + L. plantarum, 0.266 ± 0.118 vs 0.326 ± 0.009, P < 0.05). The protective effect of L. plantarum was more prominent after internal biliary drainage ( BDL + IBD vs BDL + IBD + L. plantarum, 0.415 ± 0.105 vs 0.494 ± 0.145, P < 0.05). CONCLUSION: L. plantarum can decrease intestinal epithelial cell apoptosis, reduce oxidative stress, and prevent TJ disruption in biliary obstruction by activating the PKC pathway. PMID:22912548

  7. Characterization of the locus responsible for the bacteriocin production in Lactobacillus plantarum C11.

    PubMed Central

    Diep, D B; Håvarstein, L S; Nes, I F

    1996-01-01

    Lactobacillus plantarum C11 secretes a small cationic peptide, plantaricin A, that serves as induction signal for bacteriocin production as well as transcription of plnABCD. The plnABCD operon encodes the plantaricin A precursor (PlnA) itself and determinants (PlnBCD) for a signal transducing pathway. By Northern (RNA) and sequencing analyses, four new plantaricin A-induced operons were identified. All were highly activated in concert with plnABCD upon bacteriocin induction. Two of these operons (termed plnEFI and plnJKLR) each encompass a gene pair (plnEF and plnJK, respectively) encoding two small cationic bacteriocin-like peptides with double-glycine-type leaders. The open reading frames (ORFs) encoding the bacteriocin-like peptides are followed by ORFs (plnI and -L, respectively) encoding cationic hydrophobic proteins resembling bacteriocin immunity proteins. On the third operon (termed plnMNOP), a similar bacteriocin-like ORF (plnN) and a putative immunity ORF (either plnM or -P) were identified as well. These findings suggest that two bacteriocins of two-peptide type (mature PlnEF and PlnJK) and a bacteriocin of one-peptide type (mature PlnN) could be responsible for the observed bacteriocin activity. The last operon (termed plnGHSTUV) contains two ORFs (plnGH) apparently encoding an ABC transporter and its accessory protein, respectively, known to be involved in processing and export of peptides with precursor double-glycine-type leaders. Promoter structure was established. A conserved regulatory-like box encompassing two direct repeats was identified in the promoter regions of all five plantaricin A-induced operons. These repeats may serve as regulatory elements for gene expression. PMID:8755874

  8. Optimization of growth and bacteriocin production by Lactobacillus sakei subsp. sakei2a

    PubMed Central

    Malheiros, Patrícia S.; Sant’Anna, Voltaire; Todorov, Svetoslav D.; Franco, Bernadette D.G.M.

    2015-01-01

    Lactobacillus sakei subsp. sakei 2a is a bacteriocinogenic lactic acid bacterium isolated from Brazilian pork sausage, capable of inhibiting the growth of microbial pathogens, mainly Listeria monocytogenes. In order to optimize bacteriocin production for industrial applications, this study evaluated the effect of supplementation of MRS broth with glucose, Tween 20, Tween 80, sodium citrate, potassium chloride and cysteine, and effect of the initial pH and temperature of incubation of the medium on production of bacteriocins by L. sakei 2a. Adding glucose and Tween 20 to the medium, an initial pH of 5.0 or 5.5, and incubation temperatures of 25 °C or 30 °C resulted to the highest bacteriocin yields. Thus, a 24 factorial design with the four variables was performed, and statistical analysis showed that it was an adequate model (R 2 = 0.8296). In the studied range, the four parameters significantly influenced bacteriocin production, with the maximum yield produced at an initial pH between 5.5 and 7.0, a temperature between 25 and 30 °C and supplementation of the MRS broth with glucose from 3.25 to 6.0 g L?1 and Tween 20 from 0.575 to 1.15% (v/v). Response Surface Methodology analysis indicated that the highest bacteriocin production (12800 AU mL?1) occurred in the MRS broth supplemented with 5.5 g L?1 glucose and 1.05% Tween 20 at an initial pH of 6.28 and an incubation temperature of 25 °C. The amount of bacteriocin produced in commercial MRS broths under the same conditions was only 5600AU mL?1. PMID:26413066

  9. Hypolipidemic Effects and Safety of Lactobacillus Reuteri 263 in a Hamster Model of Hyperlipidemia

    PubMed Central

    Huang, Wen-Ching; Chen, Yi-Ming; Kan, Nai-Wen; Ho, Chun-Sheng; Wei, Li; Chan, Ching-Hung; Huang, Hui-Yu; Huang, Chi-Chang

    2015-01-01

    We aimed to verify the beneficial effects of probiotic strain Lactobacillus reuteri 263 (Lr263) on hypolipidemic action in hamsters with hyperlipidemia induced by a 0.2% cholesterol and 10% lard diet (i.e., high-cholesterol diet (HCD)). Male Golden Syrian hamsters were randomly divided into two groups: normal (n = 8), standard diet (control), and experimental (n = 32), a HCD. After a two-week induction followed by a six-week supplementation with Lr263, the 32 hyperlipidemic hamsters were divided into four groups (n = 8 per group) to receive vehicle or Lr263 by oral gavage at 2.1, 4.2, or 10.5 × 109 cells/kg/day for 6 weeks, designated the HCD, 1X, 2X and 5X groups, respectively. The efficacy and safety of Lr263 supplementation were evaluated by lipid profiles of serum, liver and feces and by clinical biochemistry and histopathology. HCD significantly increased serum levels of total cholesterol (TC), triacylglycerol (TG) cholesterol, high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C), LDL-C/HDL-C ratio, hepatic and fetal TC and TG levels, and degree of fatty liver as compared with controls. Lr263 supplementation dose dependently increased serum HDL-C level and decreased serum TC, TG, LDL-C levels, LDL-C/HDL-C ratio, hepatic TC and TG levels, and fecal TG level. In addition, Lr263 supplementation had few subchronic toxic effects. Lr263 could be a potential agent with a hypolipidemic pharmacological effect. PMID:25988768

  10. Role of S-layer proteins in the biosorption capacity of lead by Lactobacillus kefir.

    PubMed

    Gerbino, Esteban; Carasi, Paula; Araujo-Andrade, Cuauhtémoc; Tymczyszyn, E Elizabeth; Gómez-Zavaglia, Andrea

    2015-04-01

    The role of S-layer proteins (SLP) on the Pb(2+) sequestrant capacity by Lactobacillus kefir CIDCA 8348 and JCM 5818 was investigated. Cultures in the stationary phase were treated with proteinase K. A dot blot assay was carried out to assess the removal of SLP. Strains with and without SLP were exposed to 0-0.5 mM Pb(NO3)2. The maximum binding capacity (q max ) and the affinity coefficient (b) were calculated using the Langmuir equation. The structural effect of Pb(2+) on microorganisms with and without SLP was determined using Raman spectroscopy. The bacterial interaction with Pb(2+) led to a broadening in the phosphate bands (1,300-1,200 cm(-1) region) and strong alterations on amide and carboxylate-related bands (?COO(-) as and ?COO(-) s). Microorganisms without SLP removed higher percentages of Pb(2+) and had higher q max than those bearing SLP. Isolated SLP had much lower q max and also removed lower percentages of Pb(2+) than the corresponding whole microorganisms. The hydrofobicity of both strains dramatically dropped when removing SLP. When bearing SLP, strains do not expose a large amount of charged groups on their surfaces, thus making less efficient the Pb(2+) removal. On the contrary, the extremely low hydrofobicity of microorganisms without SLP (and consequently, their higher capacity to remove Pb(2+)) can be explained on the basis of a greater exposure of charged chemical groups for the interaction with Pb(2+). The viability of bacteria without SLP was not significantly lower than that of bacteria bearing SLP. However, microorganisms without SLP were more prone to the detrimental effect of Pb(2+), thus suggesting that SLP acts as a protective rather than as a sequestrant layer. PMID:25653110

  11. S-Layer Protein Mediates the Stimulatory Effect of Lactobacillus helveticus MIMLh5 on Innate Immunity

    PubMed Central

    Taverniti, Valentina; Stuknyte, Milda; Minuzzo, Mario; Arioli, Stefania; De Noni, Ivano; Scabiosi, Christian; Cordova, Zuzet Martinez; Junttila, Ilkka; Hämäläinen, Sanna; Turpeinen, Hannu; Mora, Diego; Karp, Matti; Pesu, Marko

    2013-01-01

    The ability to positively affect host health through the modulation of the immune response is a feature of increasing importance in measuring the probiotic potential of a bacterial strain. However, the identities of the bacterial cell components involved in cross talk with immune cells remain elusive. In this study, we characterized the dairy strain Lactobacillus helveticus MIMLh5 and its surface-layer protein (SlpA) using in vitro and ex vivo analyses. We found that MIMLh5 and SlpA exert anti-inflammatory effects by reducing the activation of NF-?B on the intestinal epithelial Caco-2 cell line. On the contrary, MIMLh5 and SlpA act as stimulators of the innate immune system by triggering the expression of proinflammatory factors tumor necrosis factor alpha and COX-2 in the human macrophage cell line U937 via recognition through Toll-like receptor 2. In the same experiments, SlpA protein did not affect the expression of the anti-inflammatory cytokine interleukin-10. A similar response was observed following stimulation of macrophages isolated from mouse bone marrow or the peritoneal cavity. These results suggest that SlpA plays a major role in mediating bacterial immune-stimulating activity, which could help to induce the host's defenses against and responses toward infections. This study supports the concept that the viability of bacterial cells is not always essential to exert immunomodulatory effects, thus permitting the development of safer therapies for the treatment of specific diseases according to a paraprobiotic intervention. PMID:23220964

  12. Functional characterization of a mucus-specific LPXTG surface adhesin from probiotic Lactobacillus rhamnosus GG.

    PubMed

    von Ossowski, Ingemar; Satokari, Reetta; Reunanen, Justus; Lebeer, Sarah; De Keersmaecker, Sigrid C J; Vanderleyden, Jos; de Vos, Willem M; Palva, Airi

    2011-07-01

    In spite of the wealth of clinical evidence supporting the health benefits of Lactobacillus rhamnosus GG in humans, there is still a lack of understanding of the molecular mechanisms behind its probiosis. Current knowledge suggests that the health-promoting effects of this probiotic strain might be partly dependent on its persistence in the intestine and adhesion to mucosal surfaces. Moreover, L. rhamnosus GG contains mucus-binding pili that might also explain the occupation of its ecological niche as a comparatively less stringent allochthonous intestine-dwelling bacterium. To uncover additional surface proteins involved in mucosal adhesion, we investigated the adherence properties of the only predicted protein (LGG_02337) in L. rhamnosus GG that exhibits homology with a known mucus-binding domain. We cloned a recombinant form of the gene for this putative mucus adhesin and established that the purified protein readily adheres to human intestinal mucus. We also showed that this mucus adhesin is visibly distributed throughout the cell surface and participates in the adhesive interaction between L. rhamnosus GG and mucus, although less prominently than the mucus-binding pili in this strain. Based on primary structural comparisons, we concluded that the current annotation of the LGG_02337 protein likely does not accurately reflect its predicted properties, and we propose that this mucus-specific adhesin be called the mucus-binding factor (MBF). Finally, we interpret our results to mean that L. rhamnosus GG MBF, as an active mucus-specific surface adhesin with a presumed ancillary involvement in pilus-mediated mucosal adhesion, plays a part in the adherent mechanisms during intestinal colonization by this probiotic. PMID:21602388

  13. Improved 1,3-Propanediol Synthesis from Glycerol by the Robust Lactobacillus reuteri Strain DSM 20016.

    PubMed

    Ricci, Maria Antonietta; Russo, Annamaria; Pisano, Isabella; Palmieri, Luigi; de Angelis, Maria; Agrimi, Gennaro

    2015-06-01

    Various Lactobacillus reuteri strains were screened for the ability to convert glycerol to 1,3- propanediol (1,3-PDO) in a glycerol-glucose co-fermentation. Only L. reuteri DSM 20016, a well-known probiotic, was able to efficiently carry out this bioconversion. Several process strategies were employed to improve this process. CO(2+) addition to the fermentation medium, led to a high product titer (46 g/l) of 1,3-PDO and to improved biomass synthesis. L. reuteri DSM 20016 produced also ca. 3 ?g/g of cell dry weight of vitamin B12, conferring an economic value to the biomass produced in the process. Incidentally, we found that L. reuteri displays the highest resistance to CO(2+) ions ever reported for a microorganism. Two waste materials (crude glycerol from biodiesel industry and spruce hydrolysate from paper industry) alone or in combination were used as feedstocks for the production of 1,3-PDO by L. reuteri DSM 20016. Crude glycerol was efficiently converted into 1,3-PDO although with a lower titer than pure glycerol (-18%). Compared with the fermentation carried out with pure substrates, the 1,3- PDO produced was significantly lower (40.7 vs. 24.2 g/l) using cellulosic hydrolysate and crude glycerol, but strong increases of the maximal biomass produced (+27%) and of the glucose consumption rate (+46%) were found. The results of this study lay the foundation for further investigations to exploit the biotechnological potential of L. reuteri DSM 20016 to produce 1,3-PDO and vitamin B12 using industry byproducts. PMID:25588555

  14. Effects of oral intake of kimchi-derived Lactobacillus plantarum K8 lysates on skin moisturizing.

    PubMed

    Kim, Hangeun; Kim, Hye Rim; Jeong, Bong Jun; Lee, Seung Su; Kim, Tae-Rahk; Jeong, Ji Hye; Lee, Miyeong; Lee, Sinai; Lee, Jong Suk; Chung, Dae Kyun

    2015-01-01

    Skin is the soft outer covering of vertebrates that provides protection from pathogenic infection, physical damage, or UV irradiation, and controls body temperature and water content. In this study, we examined the effects of oral intake of kimchi-derived Lactobacillus plantarum K8 lysates on skin moisturizing. In an in vitro study, we observed that the hyaluronic acid content increased in HaCaT cells treated with L. plantarum K8 lysates. Oral administration of L. plantarum K8 lysates effectively attenuated the horny layer formation and decreased epidermal thickening in DNCB-treated SKH-1 hairless mice skin. The damage to barrier function was reduced after 8 weeks of oral administration of L. plantarum K8 lysates as compared with that in the atopic dermatitis mice. For the test with volunteers, we manufactured experimental candy containing 2.1% L. plantarum K8 lysates, while control candy did not contain bacterial lysate. A significant increase in hydration in the experimental candy-administered group as compared with the control candy-administered group was observed on the face after 4 and 8 weeks, and on the forearm after 4 weeks. Decreases in horny layer thickness and TEWL value were observed on the face and forearm of the experimental group. Together, the in vitro cell line and in vivo mouse studies revealed that L. plantarum K8 lysates have a moisturizing effect. A clinical research study with healthy volunteers also showed an improvement in barrier repair and function when volunteers took L. plantarum K8 lysates-containing candy. Thus, our results suggest that L. plantarum K8 lysates may help to improve skin barrier function. PMID:25179904

  15. Genomic analysis reveals Lactobacillus sanfranciscensis as stable element in traditional sourdoughs

    PubMed Central

    2011-01-01

    Sourdough has played a significant role in human nutrition and culture for thousands of years and is still of eminent importance for human diet and the bakery industry. Lactobacillus sanfranciscensis is the predominant key bacterium in traditionally fermented sourdoughs. The genome of L. sanfranciscensis TMW 1.1304 isolated from an industrial sourdough fermentation was sequenced with a combined Sanger/454-pyrosequencing approach followed by gap closing by walking on fosmids. The sequencing data revealed a circular chromosomal sequence of 1,298,316 bp and two additional plasmids, pLS1 and pLS2, with sizes of 58,739 bp and 18,715 bp, which are predicted to encode 1,437, 63 and 19 orfs, respectively. The overall GC content of the chromosome is 34.71%. Several specific features appear to contribute to the ability of L. sanfranciscensis to outcompete other bacteria in the fermentation. L. sanfranciscensis contains the smallest genome within the lactobacilli and the highest density of ribosomal RNA operons per Mbp genome among all known genomes of free-living bacteria, which is important for the rapid growth characteristics of the organism. A high frequency of gene inactivation and elimination indicates a process of reductive evolution. The biosynthetic capacity for amino acids scarcely availably in cereals and exopolysaccharides reveal the molecular basis for an autochtonous sourdough organism with potential for further exploitation in functional foods. The presence of two CRISPR/cas loci versus a high number of transposable elements suggests recalcitrance to gene intrusion and high intrinsic genome plasticity. PMID:21995419

  16. Secretory expression of a phospholipase A2 from Lactobacillus casei DSM20011 in Kluyveromyces lactis.

    PubMed

    Wang, Hui; Zhang, Liang; Shi, Guiyang

    2015-12-01

    The pla2 gene encoding a phospholipase A2 (EC 3.1.1.4) of Lactobacillus casei DSM20011 was cloned and expressed in the yeast Kluyveromyces lactis GG799 successfully for the first time. The structural pla2 gene fused in frame with the K. lactis secretion signal ?-mating factor was integrated into the LAC4 locus and expressed under the control of the LAC4 promoter. sPLA2 activity was detected in the culture supernatant during shake flask culture of K. lactis/pKLAC1-pla2. In comparison with the control strain K. lactis/pKLAC1, SDS-PAGE analysis revealed a 17-kDa recombinant protein band in K. lactis/pKLAC1-pla2, which was consistent with the predicted molecular weight of the mature protein. Real-time quantitative PCR analysis indicated that the copy number of the integrated pla2 gene ranged from 2 to 6 and positively correlated with sPLA2 activity. When the inducer galactose was used as the carbon source, the sPLA2 activity in the culture supernatant of the recombinant that harbored six pla2 gene copies reached 1.96 ± 0.15 U/mL. The influence of the culture composition and conditions on the recombinant sPLA2 activity in shake flask culture were also studied. When the recombinant was cultured at 30°C in a YPD medium culture volume of 70 mL in a 250-mL shake flask with an initial pH of 7.0, the sPLA2 activity reached 2.16 ± 0.18 U/mL. PMID:26108160

  17. Effect of Lineage-Specific Metabolic Traits of Lactobacillus reuteri on Sourdough Microbial Ecology

    PubMed Central

    Lin, Xiaoxi B.

    2014-01-01

    This study determined the effects of specific metabolic traits of Lactobacillus reuteri on its competitiveness in sourdoughs. The competitiveness of lactobacilli in sourdough generally depends on their growth rate; acid resistance additionally contributes to competitiveness in sourdoughs with long fermentation times. Glycerol metabolism via glycerol dehydratase (gupCDE) accelerates growth by the regeneration of reduced cofactors; glutamate metabolism via glutamate decarboxylase (gadB) increases acid resistance by generating a proton motive force. Glycerol and glutamate metabolisms are lineage-specific traits in L. reuteri; therefore, this study employed glycerol dehydratase-positive sourdough isolates of human-adapted L. reuteri lineage I, glutamate decarboxylase-positive strains of rodent-adapted L. reuteri lineage II, as well as mutants with deletions in gadB or gupCDE. The competitivenesses of the strains were quantified by inoculation of wheat and sorghum sourdoughs with defined strains, followed by propagation of doughs with a 10% inoculum and 12-h or 72-h fermentation cycles. Lineage I L. reuteri strains dominated sourdoughs propagated with 12-h fermentation cycles; lineage II L. reuteri strains dominated sourdoughs propagated with 72-h fermentation cycles. L. reuteri 100-23?gadB was outcompeted by its wild-type strain in sourdoughs fermented with 72-h fermentation cycles; L. reuteri FUA3400?gupCDE was outcompeted by its wild-type strain in sourdoughs fermented with both 12-h and 72-h fermentation cycles. Competition experiments with isogenic pairs of strains resulted in a constant rate of strain displacement of the less competitive mutant strain. In conclusion, lineage-specific traits of L. reuteri determine the competitiveness of this species in sourdough fermentations. PMID:25015888

  18. Lactobacillus amylovorus DSM 19280 as a novel food-grade antifungal agent for bakery products.

    PubMed

    Ryan, Liam A M; Zannini, Emanuele; Dal Bello, Fabio; Pawlowska, Agata; Koehler, Peter; Arendt, Elke K

    2011-04-29

    Mould spoilage is the main cause of substantial economic loss in bakery industry and might also cause public health problems due to the production of mycotoxins. The reduction of mould growth in bakery products is thus of crucial importance and there is great interest to develop safe and efficient strategies for this purpose. In this study Lactobacillus amylovorus DSM19280 has been shown to produce a wide spectrum of antifungal compounds active against common bread spoilage fungi. Among the indicator moulds, Aspergillus fumigatus and Fusarium culmorum were the most sensitive organisms. Several antifungal compounds were found to be present in synthetic medium inoculated with L. amylovorus DSM19280 strain, some of them being reported here for the first time. Wheat doughs fermented with L. amylovorus DSM19280 had good rheological properties and the breads thereof were of high quality as shown by rheofermentometer and texture analyser measurements. The results were compared with those obtained with a control non-antifungal L. amylovorus DSM20531(T) strain, a non-acidified and a chemically acidified dough. The quality of sourdough and bread fermented with L. amylovorus DSM 19280 was comparable to that obtained by using L. amylovorus DSM20531 (T). Additionally, breads were evaluated for the ability to retard the growth of Fusarium culmorum FST 4.05, Aspergillus niger FST4.21, Penicillium expansum FST 4.22, Penicillium roqueforti FST 4.11 and fungal flora from the bakery environment. The biological preservation of bread with L. amylovorus DSM 19280 was also compared to the most commonly used antifungal agent Calcium propionate. Breads containing sourdough fermented with L. amylovorus DSM 19280 were more effective in extending the shelf life of bread than the calcium propionate. PMID:21429613

  19. Performance of stress resistant variants of Listeria monocytogenes in mixed species biofilms with Lactobacillus plantarum.

    PubMed

    Metselaar, Karin I; Saá Ibusquiza, Paula; Ortiz Camargo, Angela R; Krieg, Myriam; Zwietering, Marcel H; den Besten, Heidy M W; Abee, Tjakko

    2015-11-20

    Population diversity and the ability to adapt to changing environments allow Listeria monocytogenes to grow and survive under a wide range of environmental conditions. In this study, we aimed to evaluate the performance of a set of acid resistant L. monocytogenes variants in mixed-species biofilms with Lactobacillus plantarum as well as their benzalkonium chloride (BAC) resistance in these biofilms. L. monocytogenes LO28 wild type and acid resistant variants were capable of forming mixed biofilms with L. plantarum at 20°C and 30°C in BHI supplemented with manganese and glucose. Homolactic fermentation of glucose by L. plantarum created an acidic environment with pH values below the growth boundary of L. monocytogenes. Some of the variants were able to withstand the low pH in the mixed biofilms for a longer time than the WT and there were clear differences in survival between the variants which could not be correlated to (lactic) acid resistance alone. Adaptation to mild pH of liquid cultures during growth to stationary phase increased the acid resistance of some variants to a greater extent than of others, indicating differences in adaptive behaviour between the variants. Two variants that showed a high level of acid adaptation when grown in liquid cultures, showed also better performance in mixed species biofilms. There were no clear differences in BAC resistance between the wild type and variants in mixed biofilms. It can be concluded that acid resistant variants of L. monocytogenes show diversity in their adaptation to acidic conditions and their capacity to survive in mixed cultures and biofilms with L. plantarum. PMID:25935090

  20. Lactobacillus fermentum NCIMB 5221 and NCIMB 2797 as cholesterol-lowering probiotic biotherapeutics: in vitro analysis.

    PubMed

    Tomaro-Duchesneau, C; Saha, S; Malhotra, M; Jones, M L; Rodes, L; Prakash, S

    2015-12-01

    Cardiovascular and coronary artery disease risk are correlated with cholesterol levels and are significant health concerns. Current cholesterol-lowering approaches includes lifestyle and diet modifications, as well as statins which presents numerous shortcomings. The probiotic bacteria, Lactobacillus fermentum NCIMB 5221 and NCIMB 2797, have demonstrated cholesterol-lowering potential in animal studies. However, there is a lack in understanding the mechanism(s) behind these observed effects. The goal of this work is to investigate, in vitro, the cholesterol-lowering mechanisms of these two strains. To determine the cholesterol-lowering mechanisms, probiotic cholesterol assimilation, colon epithelial adhesion and inhibition of cholesterol uptake by colon epithelial (Caco-2) cells were investigated. L. fermentum NCIMB 2797 (P=0.012) and NCIMB 5221 (P=0.003) assimilated cholesterol and their cell surface hydrophobicity was 70.30±8.85% and 55.60±2.59%, respectively. Both L. fermentum strains showed no significant impact (P>0.05) on Caco-2 cell viability. Of most interest, Caco-2 pre-exposure to L. fermentum NCIMB 5221 significantly decreased (P=0.015) cholesterol uptake, with 85.98±2.07% uptake compared to the untreated cells. Similarly, L. fermentum NCIMB 2797 probiotic cells significantly decreased (P=0.019) cholesterol uptake by Caco-2 cells, with 86.45±1.71% uptake observed compared to the control cells. The results demonstrate that L. fermentum NCIMB 5221 and L. fermentum NCIMB 2797 have the potential via various modes of action to lower cholesterol. Additional studies are required to understand the mechanism(s) of action behind probiotic cholesterol assimilation and behind the cholesterol uptake inhibition by colon epithelial cells. PMID:26322545

  1. Multi-Cereal Beverage Fermented by Lactobacillus Helveticus and Saccharomyces Cerevisiae.

    PubMed

    Ai, Jing; Li, Ai-Li; Su, Ben-Xian; Meng, Xiang-Chen

    2015-06-01

    A novel multi-cereal-based fermented beverage with suitable aroma, flavor, and pH fermented by lactic acid bacteria and Saccharomyces cerevisiae was developed. Twenty-seven lactobacilli strains were screened for acid production (pH and titratable acidity) in a mixture of malt, rice, and maize substrates. It was found that Lactobacillus helveticus KLDS1.9204 had the greatest acid production among 27 lactobacilli tested. The fermentation performance of L. helveticus KLDS1.9204 was also assayed and the fermentation parameters were optimized using Plackett-Burman design and steepest ascent method. L. helveticus KLDS1.9204 showed good proteolytic capability, however, the strain could not utilize starch. The optimum substrate consisted of 50% malt (25 g/100 mL), 25% rice (20 g/100 mL), and 25% maize (30 g/100 mL). The inoculum was 5% with a ratio of S. cerevisiae to L. helveticus KLDS1.9204 of 2.5:1. The optimum temperature was 37 °C and the time was 22 h. Lastly, the quality of the multi-cereal-based fermented beverage was evaluated. This beverage was light yellow, transparent, and it tasted well with a pleasant acid and a unique flavor of cereals. The beverage was rich in free amino acids and organic acids. The pH and titratable acidity of the beverage were 3.5 and 29.86 °T, respectively. The soluble solids content of the beverage was 6.5 °Brix, and the alcohol content was 0.67%. PMID:25962443

  2. Ehealth: Low FODMAP diet vs Lactobacillus rhamnosus GG in irritable bowel syndrome

    PubMed Central

    Pedersen, Natalia; Andersen, Nynne Nyboe; Végh, Zsuzsanna; Jensen, Lisbeth; Ankersen, Dorit Vedel; Felding, Maria; Simonsen, Mette Hestetun; Burisch, Johan; Munkholm, Pia

    2014-01-01

    AIM: To investigate the effects of a low fermentable, oligosaccharides, disaccharides, monosaccharides and polyols diet (LFD) and the probiotic Lactobacillus rhamnosus GG (LGG) in irritable bowel syndrome (IBS). METHODS: Randomised, unblinded controlled trial on the effect of 6-wk treatment with LFD, LGG or a normal Danish/Western diet (ND) in patients with IBS fulfilling Rome III diagnostic criteria, recruited between November 2009 and April 2013. Patients were required to complete on a weekly basis the IBS severity score system (IBS-SSS) and IBS quality of life (IBS-QOL) questionnaires in a specially developed IBS web self-monitoring application. We investigated whether LFD or LGG could reduce IBS-SSS and improve QOL in IBS patients. RESULTS: One hundred twenty-three patients (median age 37 years, range: 18-74 years), 90 (73%) females were randomised: 42 to LFD, 41 to LGG and 40 to ND. A significant reduction in mean ± SD of IBS-SSS from baseline to week 6 between LFD vs LGG vs ND was revealed: 133 ± 122 vs 68 ± 107, 133 ± 122 vs 34 ± 95, P < 0.01. Adjusted changes of IBS-SSS for baseline covariates showed statistically significant reduction of IBS-SSS in LFD group compared to ND (IBS-SSS score 75; 95%CI: 24-126, P < 0.01), but not in LGG compared to ND (IBS-SSS score 32; 95%CI: 18-80, P = 0.20). IBS-QOL was not altered significantly in any of the three groups: mean ± SD in LFD 8 ± 18 vs LGG 7 ± 17, LFD 8 ± 18 vs ND 0.1 ± 15, P = 0.13. CONCLUSION: Both LFD and LGG are efficatious in patients with IBS. PMID:25473176

  3. Beneficial Effects of Citrus Juice Fermented with Lactobacillus plantarum YIT 0132 on Japanese Cedar Pollinosis

    PubMed Central

    HARIMA-MIZUSAWA, Naomi; IINO, Tohru; ONODERA-MASUOKA, Norie; KATO-NAGAOKA, Noriko; KIYOSHIMA-SHIBATA, Junko; GOMI, Atsushi; SHIBAHARA-SONE, Harue; KANO, Mitsuyoshi; SHIDA, Kan; SAKAI, Masashi; MIYAZAKI, Kouji; ISHIKAWA, Fumiyasu

    2014-01-01

    Recently, the prevalence of allergies in Japan has been increasing. Certain types of fruit juice and lactic acid bacteria are known to alleviate allergic symptoms. Therefore, we examined whether citrus juice fermented by a specific lactic acid bacteria can improve the symptoms of Japanese cedar pollinosis (JCPsis). Lactobacillus plantarum YIT 0132 (LP0132) was selected based on its high proliferative activity in citrus juice and anti-inflammatory interleukin-10-inducing activity. Dietary administration of heat-killed LP0132 cells or citrus juice fermented with LP0132 was found to significantly suppress nasal rubbing in a JCPsis mouse model, indicating relief of allergy symptoms. To evaluate the effects of LP0132-fermented citrus juice on pollinosis symptoms and quality of life (QOL) in humans with JCPsis, a single-blind, placebo-controlled, parallel-group clinical trial was conducted. The participants were 42 adults with JCPsis. They ingested 100?mL of sterilized LP0132-fermented citrus juice (active group) or unfermented citrus juice (placebo group) once daily for 8 weeks. Immediately after the pollen peak when allergy symptoms and QOL loss were most severe, itchy eyes, itchy skin, and QOL loss by JCPsis were alleviated in the active group compared with the placebo group. At 10 weeks after starting the intervention, increased the levels of blood eosinophils were significantly suppressed in the active group compared with the placebo group. We conclude that continuous ingestion of citrus juice fermented with LP0132 may help alleviate the allergy symptoms and impaired QOL caused by JCPsis. PMID:25379362

  4. Identification of the transcriptional response of human intestinal mucosa to Lactobacillus plantarum WCFS1 in vivo

    PubMed Central

    Troost, Freddy J; van Baarlen, Peter; Lindsey, Patrick; Kodde, Andrea; de Vos, Willem M; Kleerebezem, Michiel; Brummer, Robert-Jan M

    2008-01-01

    Background There is limited knowledge on the extent and dynamics of the mucosal response to commensal and probiotic species in the human intestinal lumen. This study aimed to identify the acute, time-dependent responses of intestinal mucosa to commensal Lactobacillus plantarum WCFS1 in vivo in two placebo-controlled human intervention studies in healthy volunteers. Transcriptional changes in duodenal mucosa upon continuous intraduodenal infusion of L. plantarum WCFS1 for one- and six h, respectively, were studied using oro- and nasogastric intubations with dedicated orogastric catheters and tissue sampling by standard flexible gastroduodenoscopy. Results One- and six-h exposure of small intestinal mucosa to L. plantarum WCFS1 induced differential expression of 669 and 424 gene reporters, respectively. While short-term exposure to L. plantarum WCFS1 inhibited fatty acid metabolism and cell cycle progression, cells switched to a more proliferative phase after prolonged exposure with an overall expression profile characterized by upregulation of genes involved in lipid metabolism, cellular growth and development. Cell death and immune responses were triggered, but cell death-executing genes or inflammatory signals were not expressed. Proteome analysis showed differential expression of several proteins. Only the microsomal protein 'microsomal triglyceride transfer protein' was regulated on both the transcriptional and the protein level in all subjects. Conclusion Overall, this study showed that intestinal exposure to L. plantarum WCFS1 induced consistent, time-dependent transcriptional responses in healthy intestinal mucosa. This extensive exploration of the human response to L. plantarum WCFS1 could eventually provide molecular support for specific or probiotic activity of this strain or species, and exemplifies the strength of the applied technology to identify the potential bio-activity of microbes in the human intestine. PMID:18681965

  5. The Evolution of Host Specialization in the Vertebrate Gut Symbiont Lactobacillus reuteri

    SciTech Connect

    Frese, Steven A.; Benson, Andrew K.; Tannock, Gerald W.; Loach, Diane M.; Kim, Jaehyoung; Zhang, Min; Oh, Phaik Lyn; Heng, Nicholas C. K.; Patil, Prabhu; Juge, Nathalie; MacKenzie, Donald A.; Pearson, Bruce M.; Lapidus, Alla L.; Dalin, Eileen; Tice, Hope; Goltsman, Eugene; Land, Miriam L; Hauser, Loren John; Ivanova, N; Kyrpides, Nikos C; Walter, Jens

    2011-01-01

    Recent research has provided mechanistic insight into the important contributions of the gut microbiota to vertebrate biology, but questions remain about the evolutionary processes that have shaped this symbiosis. In the present study, we showed in experiments with gnotobiotic mice that the evolution of Lactobacillus reuteri with rodents resulted in the emergence of host specialization. To identify genomic events marking adaptations to the murine host, we compared the genome of the rodent isolate L. reuteri 100-23 with that of the human isolate L. reuteri F275, and we identified hundreds of genes that were specific to each strain. In order to differentiate true host-specific genome content from strain-level differences, comparative genome hybridizations were performed to query 57 L. reuteri strains originating from six different vertebrate hosts in combination with genome sequence comparisons of nine strains encompassing five phylogenetic lineages of the species. This approach revealed that rodent strains, although showing a high degree of genomic plasticity, possessed a specific genome inventory that was rare or absent in strains from other vertebrate hosts. The distinct genome content of L. reuteri lineages reflected the niche characteristics in the gastrointestinal tracts of their respective hosts, and inactivation of seven out of eight representative rodent-specific genes in L. reuteri 100-23 resulted in impaired ecological performance in the gut of mice. The comparative genomic analyses suggested fundamentally different trends of genome evolution in rodent and human L. reuteri populations, with the former possessing a large and adaptable pan-genome while the latter being subjected to a process of reductive evolution. In conclusion, this study provided experimental evidence and a molecular basis for the evolution of host specificity in a vertebrate gut symbiont, and it identified genomic events that have shaped this process.

  6. Polyphosphate, an active molecule derived from probiotic Lactobacillus brevis, improves the fibrosis in murine colitis.

    PubMed

    Kashima, Shin; Fujiya, Mikihiro; Konishi, Hiroaki; Ueno, Nobuhiro; Inaba, Yuhei; Moriichi, Kentaro; Tanabe, Hiroki; Ikuta, Katsuya; Ohtake, Takaaki; Kohgo, Yutaka

    2015-08-01

    Inflammatory bowel disease frequently causes intestinal obstruction because of extensive fibrosis. This study investigated whether polyphosphate (poly P), an active molecule derived from Lactobacillus brevis, could improve the fibrosis in a model of chronic colitis. In this study, dextran sodium sulfate (DSS)-induced chronic colitis models and trinitrobenzene sulfonic acid (TNBS)-induced colitis models were used as models of fibrosis. To clarify the mechanism responsible for the observed effects, Caco-2/brush border epithelial (BBE) and naive T helper lymphocyte (THP)-1 cells were treated with lipopolysaccharide (LPS) to induce inflammation. Non-cancer human colon fibroblast (CCD-18) cells were treated with transforming growth factor beta 1 (TGF-?1) to induce fibrosis. The expression levels of fibrosis- and inflammation-associated molecules were evaluated by both a Western blotting analysis and reverse transcriptase-polymerase chain reaction (RT-PCR). The histologic inflammation and fibrosis were significantly improved in the group administered poly P in both the DSS and TNBS colitis models. The levels of interleukin 1? (IL-1?) and tumor necrosis factor ? (TNF-?) were significantly decreased by poly P treatment. The expression levels of TGF-?1 and collagens in the colitis mice were decreased by poly P. The LPS-induced expressions of IL-1? and TGF-?1 in Caco-2/BBE cells and of TNF-? in THP-1 cells were reduced by poly P treatment. Poly P did not affect the expression of collagens and connective tissue growth factor in the CCD-18 cells. In conclusion, poly P suppresses intestinal inflammation and fibrosis by downregulating the expression of inflammation- and fibrosis-associated molecules in the intestinal epithelium. The administration of poly P is therefore a novel option to treat fibrosis because of chronic intestinal inflammation. PMID:25766132

  7. Role of Plasmids in Lactobacillus brevis BSO 464 Hop Tolerance and Beer Spoilage

    PubMed Central

    Bergsveinson, Jordyn; Baecker, Nina; Pittet, Vanessa

    2014-01-01

    Specific isolates of lactic acid bacteria (LAB) can grow in the harsh beer environment, thus posing a threat to brew quality and the economic success of breweries worldwide. Plasmid-localized genes, such as horA, horC, and hitA, have been suggested to confer hop tolerance, a trait required for LAB survival in beer. The presence and expression of these genes among LAB, however, do not universally correlate with the ability to grow in beer. Genome sequencing of the virulent beer spoilage organism Lactobacillus brevis BSO 464 revealed the presence of eight plasmids, with plasmids 1, 2, and 3 containing horA, horC, and hitA, respectively. To investigate the roles that these and the other five plasmids play in L. brevis BSO 464 growth in beer, plasmid curing with novobiocin was used to derive 10 plasmid variants. Multiplex PCRs were utilized to determine the presence or absence of each plasmid, and how plasmid loss affected hop tolerance and growth in degassed (noncarbonated) beer was assessed. Loss of three of the eight plasmids was found to affect hop tolerance and growth in beer. Loss of plasmid 2 (horC and 28 other genes) had the most dramatic effect, with loss of plasmid 4 (120 genes) and plasmid 8 (47 genes) having significant, but smaller, impacts. These results support the contention that genes on mobile genetic elements are essential for bacterial growth in beer and that beer spoilage ability is not dependent solely on the three previously described hop tolerance genes or on the chromosome of a beer spoilage LAB isolate. PMID:25501474

  8. Genome Sequence and Comparative Genome Analysis of Lactobacillus casei: Insights into Their Niche-Associated Evolution

    PubMed Central

    Cai, Hui; Thompson, Rebecca; Budinich, Mateo F.; Broadbent, Jeff R.

    2009-01-01

    Lactobacillus casei is remarkably adaptable to diverse habitats and widely used in the food industry. To reveal the genomic features that contribute to its broad ecological adaptability and examine the evolution of the species, the genome sequence of L. casei ATCC 334 is analyzed and compared with other sequenced lactobacilli. This analysis reveals that ATCC 334 contains a high number of coding sequences involved in carbohydrate utilization and transcriptional regulation, reflecting its requirement for dealing with diverse environmental conditions. A comparison of the genome sequences of ATCC 334 to L. casei BL23 reveals 12 and 19 genomic islands, respectively. For a broader assessment of the genetic variability within L. casei, gene content of 21 L. casei strains isolated from various habitats (cheeses, n = 7; plant materials, n = 8; and human sources, n = 6) was examined by comparative genome hybridization with an ATCC 334-based microarray. This analysis resulted in identification of 25 hypervariable regions. One of these regions contains an overrepresentation of genes involved in carbohydrate utilization and transcriptional regulation and was thus proposed as a lifestyle adaptation island. Differences in L. casei genome inventory reveal both gene gain and gene decay. Gene gain, via acquisition of genomic islands, likely confers a fitness benefit in specific habitats. Gene decay, that is, loss of unnecessary ancestral traits, is observed in the cheese isolates and likely results in enhanced fitness in the dairy niche. This study gives the first picture of the stable versus variable regions in L. casei and provides valuable insights into evolution, lifestyle adaptation, and metabolic diversity of L. casei. PMID:20333194

  9. Kinetic studies on exploring lactose hydrolysis potential of ? galactosidase extracted from Lactobacillus plantarum HF571129.

    PubMed

    Selvarajan, E; Mohanasrinivasan, V

    2015-10-01

    A novel intracellular ?-galactosidases produced by Lactobacillus plantarum HF571129, isolated from an Indian traditional fermented milk product curd was purified and characterized. The ?-galactosidases is a hetrodimer with a molecular weight of 60 kDa (larger subunit) and 42 kDa (smaller subunit), as estimated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme was purified 7.23 fold by ultrasonication, ultrafiltration and gel filtration chromatography with an overall recovery of 30.41 %. The optimum temperature for hydrolysis of its preferred substrates, o-nitrophenyl- ?-D-galactopyranoside (ONPG) and lactose, are 50 °C (both), and optimum pH for these reactions is 6.5 and 7.5, respectively. The ?-galactosidases showed higher affinity for ONPG (Km, 6.644 mM) as compared to lactose (Km, 23.28 mM). Galactose, the end product of lactose hydrolysis was found to be inhibited (47 %). The enzyme activity was drastically altered by the metal ion chelators EDTA, representing that this enzyme is a metalloenzyme. The enzyme was activated to a larger extent by Mg(2+) (73 % at 1 mM), while inhibited at higher concentrations of Na(+) (54 % at 100 mM), K(+) (16 % at 100 mM) and urea (16 % at 100 mM). The thermal stability study indicated an inactivation energy of Ed?=?171.37 kJ mol(-1). Thermodynamic parameters such as ?H, ?S and ?G, were determined as a function of temperature. About 88 % of lactose was hydrolyzed at room temperature within 1 h. The study suggested that this enzyme showed its obvious superiority in the industrial lactose conversion process. PMID:26396367

  10. Lactobacillus GG and Tributyrin Supplementation Reduce Antibiotic-Induced Intestinal Injury

    PubMed Central

    Cresci, Gail; Nagy, Laura E.; Ganapathy, Vadivel

    2013-01-01

    Background Antibiotic therapy negatively alters the gut microbiota. Lactobacillus GG (LGG) decreases antibiotic-associated diarrhea (AAD) symptoms, but the mechanisms are unknown. Butyrate has beneficial effects on gut health. Altered intestinal gene expression occurs in the absence of gut microbiota. We hypothesized that antibiotic-induced changes in gut microbiota reduce butyrate production, varying genes involved with gut barrier integrity and water and electrolyte absorption, lending to AAD, and that simultaneous supplementation with LGG and/or tributyrin would prevent these changes. Methods C57BL/6 mice aged 6–8 weeks received a chow diet while divided into 8 treatment groups (± saline, ± LGG, ± tributyrin, or both). Mice received treatments orally for 7 days with ± broad-spectrum antibiotics. Water intake was recorded daily and body weight was measured. Intestine tissue samples were obtained and analyzed for expression of genes and proteins involved with water and electrolyte absorption, butyrate transport, and gut integrity via polymerase chain reaction and immunohistochemistry. Results Antibiotics decreased messenger RNA (mRNA) expression (butyrate transporter and receptor, Na+/H+ exchanger, Cl?/HCO3?, and a water channel) and protein expression (butyrate transporter, Na+/H+ exchanger, and tight junction proteins) in the intestinal tract. LGG and/or tributyrin supplementation maintained intestinal mRNA expression to that of the control animals, and tributyrin maintained intestinal protein intensity expression to that of control animals. Conclusion Broad-spectrum antibiotics decrease expression of anion exchangers, butyrate transporter and receptor, and tight junction proteins in mouse intestine. Simultaneous oral supplementation with LGG and/or tributyrin minimizes these losses. Optimizing intestinal health with LGG and/or tributyrin may offer a preventative therapy for AAD. PMID:23630018

  11. A novel Lactobacillus plantarum strain P-8 activates beneficial immune response of broiler chickens.

    PubMed

    Wang, Lifeng; Liu, Caihong; Chen, Ma; Ya, Tuo; Huang, Weiqiang; Gao, Pengfei; Zhang, Heping

    2015-12-01

    To investigate whether Lactobacillus plantarum P-8 may be used as an alternative to antibiotics in the broiler chicken diet, we compared P-8 and antibiotics for their immunobiotic properties and their effect on growth performance of broiler chickens in a 42-day trial. The results showed that P-8 provided similar benefits in weight gain, feed intake and feed efficiency as antibiotics did. Importantly, P-8 activated protective immune responses of the broilers while antibiotics lacked this effect. P-8 induced higher fecal secretory IgA (sIgA) levels on day 42 (P?0.027) and IgA(+) lymphocytes in the jejunum and Peyer's patches (PP) (P<0.001) compared to antibiotic treatment. Antibiotics reduced the IgA(+) lymphocytes in jejunum and PP on day 42 compared to the control. P-8 increased CD3(+) T cells in the small intestinal tissues in most test situations whereas antibiotics had fewer CD3(+) cells in PP and cecal tonsil compared with the control broilers at the end of the trial. In addition, P-8 increased CD4(+) T cells significantly in the intestinal tissues compared to both antibiotics and the control (P<0.0052). Both Th1 and Th2 cytokine expression were enhanced by P-8 on day 14, consistent with the clinical trial results showing probiotic benefits in diseases. Antibiotics up- and down-regulated interleukin (IL)-2, IL-4 and IL-10 transcripts in an age-dependent manner, and showed anti-inflammatory potential. These data indicate that P-8 may provide protective immune response to broilers while maintaining similar growth performance and may be a potential alternative to antibiotics supplemented in chicken feeds. PMID:26481964

  12. Studies on identifying the binding sites of folate and its derivatives in Lactobacillus casei thymidylate synthase

    SciTech Connect

    Maley, F.; Maley, G.F.

    1983-01-01

    It was shown that folate and its derivatives have a profound effect on stabilizing thymidylate synthase in vitro and in vivo, as a consequence of ternary formation between the folate, dUMP, or FdUMP, and the synthase. The degree to which complex formation is affected can be revealed qualitatively by circular dichroism and quantitatively by equilibrium dialysis using the Lactobacillus casei synthase. In contrast to the pteroylmonoglutamates, the pteroylpolyglutamates bind to thymidylate synthase in the absence of dUMP, but even their binding affinity is increased greatly by this nucleotide or its analogues. Similarly, treatment of the synthase with carboxypeptidase A prevents the binding of the pteroylmonoglutamates and reduces the binding of the polyglutamates without affecting dUMP binding. The latter does not protect against carboxypeptidase inactivation but does potentiate the protective effect of the pteroylpolyglutamates. To determine the region of the synthase involved in the binding of the glutamate residues, Pte(/sup 14/C)GluGlu6 was activated by a water soluble carbodiimide in the presence and absence of dUMP. This folate derivative behaved as a competitive inhibitor of 5,10-CH/sub 2/H/sub 4/PteGlu, in contrast to methotrexate which was non-competitive. Separation of the five cyanogen bromide peptides from the L. casei synthase revealed 80% of the radioactivity to be associated with CNBr-2 and about 15% with CNBr-4. Chymotrypsin treatment of CNBr-2 yielded two /sup 14/C-labeled peaks on high performance liquid chromatography, with the slower migrating one being separated further into two peaks by Bio-gel P2 chromatography. All three peptides came from the same region of CNBr-2, encompassing residues 47-61 of the enzyme. From these studies it would appear that the residues most probably involved in the fixation of PteGlu7 are lysines 50 and 58. In contrast, methotrexate appeared to bind to another region of CNBr-2.

  13. A study of the thermal denaturation of the S-layer protein from Lactobacillus salivarius

    NASA Astrophysics Data System (ADS)

    Lighezan, Liliana; Georgieva, Ralitsa; Neagu, Adrian

    2012-09-01

    Surface layer (S-layer) proteins display an intrinsic self-assembly property, forming monomolecular crystalline arrays, identified in outermost structures of the cell envelope in many organisms, such as bacteria and archaea. Isolated S-layer proteins also possess the ability to recrystallize into regular lattices, being used in biotechnological applications, such as controlling the architecture of biomimetic surfaces. To this end, the stability of the S-layer proteins under high-temperature conditions is very important. In this study, the S-layer protein has been isolated from Lactobacillus salivarius 16 strain of human origin, and purified by cation-exchange chromatography. Using circular dichroism (CD) spectroscopy, we have investigated the thermal denaturation of the S-layer protein. The far- and near-UV CD spectra have been collected, and the temperature dependence of the CD signal in these spectral domains has been analyzed. The variable temperature results show that the secondary and tertiary structures of the S-layer protein change irreversibly due to the heating of the sample. After the cooling of the heated protein, the secondary and tertiary structures are partially recovered. The denaturation curves show that the protein unfolding depends on the sample concentration and on the heating rate. The secondary and tertiary structures of the protein suffer changes in the same temperature range. We have also detected an intermediate state in the protein denaturation pathway. Our results on the thermal behavior of the S-layer protein may be important for the use of S-layer proteins in biotechnological applications, as well as for a better understanding of the structure and function of S-layer proteins.

  14. Metabolic Responses of Lactobacillus plantarum Strains during Fermentation and Storage of Vegetable and Fruit Juices

    PubMed Central

    Filannino, P.; Cardinali, G.; Rizzello, C. G.; Buchin, S.; De Angelis, M.; Gobbetti, M.

    2014-01-01

    Strains of Lactobacillus plantarum were grown and stored in cherry (ChJ), pineapple (PJ), carrot (CJ), and tomato (TJ) juices to mimic the chemical composition of the respective matrices. Wheat flour hydrolysate (WFH), whey milk (W), and MRS broth were also used as representatives of other ecosystems. The growth rates and cell densities of L. plantarum strains during fermentation (24 h at 30°C) and storage (21 days at 4°C) differed only in part, being mainly influenced by the matrix. ChJ and PJ were the most stressful juices for growth and survival. Overall, the growth in juices was negatively correlated with the initial concentration of malic acid and carbohydrates. The consumption of malic acid was noticeable for all juices, but mainly during fermentation and storage of ChJ. Decreases of branched-chain amino acids (BCAA)—with the concomitant increase of their respective branched alcohols—and His and increases of Glu and gamma-aminobutyric acid (GABA) were the main traits of the catabolism of free amino acids (FAA), which were mainly evident under less acidic conditions (CJ and TJ). The increase of Tyr was found only during storage of ChJ. Some aldehydes (e.g., 3-methyl-butanal) were reduced to the corresponding alcohols (e.g., 3-methyl-1-butanol). After both fermentation and storage, acetic acid increased in all fermented juices, which implied the activation of the acetate kinase route. Diacetyl was the ketone found at the highest level, and butyric acid increased in almost all fermented juices. Data were processed through multidimensional statistical analyses. Except for CJ, the juices (mainly ChJ) seemed to induce specific metabolic traits, which differed in part among the strains. This study provided more in-depth knowledge on the metabolic mechanisms of growth and maintenance of L. plantarum in vegetable and fruit habitats, which also provided helpful information to select the most suitable starters for fermentation of targeted matrices. PMID:24487533

  15. Antifungal activity of Lactobacillus against Microsporum canis, Microsporum gypseum and Epidermophyton floccosum

    PubMed Central

    Guo, Jiahui; Brosnan, Brid; Furey, Ambrose; Arendt, Elke; Murphy, Padraigin; Coffey, Aidan

    2012-01-01

    A total of 220 lactic acid bacteria isolates were screened for antifungal activity using Aspergillus fumigatus and Aspergillus niger as the target strains. Four Lactobacillus strains exhibited strong inhibitory activity on agar surfaces. All four were also identified as having strong inhibitory activity against the human pathogenic fungi Microsporum canis, Microsporum gypseum and Epidermophyton floccosum. One of the four lactobacilli, namely Lb. reuteri ee1p exhibited the most inhibition against dermatophytes. Cell-free culture supernatants of Lb. reuteri ee1p and of the non-antifungal Lb. reuteri M13 were freeze-dried and used to access and compare antifungal activity in agar plate assays and microtiter plate assays. Addition of the Lb. reuteri ee1p freeze-dried cell-free supernatant powder into the agar medium at concentrations greater than 2% inhibited all fungal colony growth. Addition of the powder at 5% to liquid cultures caused complete inhibition of fungal growth on the basis of turbidity. Freeze-dried supernatant of the non-antifungal Lb. reuteri M13 at the same concentrations had a much lesser effect. As Lb. reuteri M13 is very similar to the antifungal strain ee1p in terms of growth rate and final pH in liquid culture, and as it has little antifungal activity, it is clear that other antifungal compounds must be specifically produced (or produced at higher levels) by the anti-dermatophyte strain Lb. reuteri ee1p. Reuterin was undetectable in all four antifungal strains. The cell free supernatant of Lb. reuteri ee1p was analyzed by LC-FTMS using an Accela LC coupled to an LTQ Orbitrap XL mass spectrometer. The high mass accuracy spectrum produced by compounds in the Lb. reuteri ee1p strain was compared with both a multianalyte chromatogram and individual spectra of standard anti-fungal compounds, which are known to be produced by lactic acid bacteria. Ten antifungal metabolites were detected. PMID:22539027

  16. Metabolic Engineering of Lactobacillus plantarum for Production of l-Ribulose?

    PubMed Central

    Helanto, M.; Kiviharju, K.; Leisola, M.; Nyyssölä, A.

    2007-01-01

    l-Ribulose is a rare and expensive sugar that can be used as a precursor for the production of other rare sugars of high market value such as l-ribose. In this work we describe a production process for l-ribulose using l-arabinose, a common component of polymers of lignocellulosic materials, as the starting material. A ribulokinase-deficient mutant of the heterofermentative lactic acid bacterium Lactobacillus plantarum NCIMB8826 was constructed. Expression of araA, which encodes the critical enzyme l-arabinose isomerase, was repressed by high glucose concentrations in batch cultivations. A fed-batch cultivation strategy was therefore used to maximize l-arabinose isomerase production during growth. Resting cells of the ribulokinase-deficient mutant were used for the production of l-ribulose. The isomerization of l-arabinose to l-ribulose was very unfavorable for l-ribulose formation. However, high l-ribulose yields were obtained by complexing the produced l-ribulose with borate. The process for l-ribulose production in borate buffer by resting cells was optimized using central composite designs. The experiment design suggested that the process has an optimal operation point around an l-arabinose concentration of 100 g liter?1, a borate concentration of 500 mM, and a temperature of 48°C, where the statistical software predicted an initial l-ribulose production rate of 29.1 g liter?1 h?1, a best-achievable process productivity of 14.8 g liter?1 h?1, and a conversion of l-arabinose to l-ribulose of 0.70 mol mol?1. PMID:17873078

  17. Preventive Effect of Lactobacillus fermentum Zhao on Activated Carbon-Induced Constipation in Mice.

    PubMed

    Zhao, Xin; Qian, Yu; Suo, Huayi; Du, Muying; Li, Guijie; Liu, Zhenhu; Li, Jian

    2015-01-01

    The aim of this study was to investigate the effects of Lactobacillus fermentum Zhao (LF-Zhao) on activated carbon-induced constipation in ICR mice. ICR mice were administered lactic acid bacteria by gavage for 9 d. Body weight, diet intake, drinking amount, stool status, gastrointestinal transit distance and stool time, in addition to motilin (MTL), gastrin (Gas), endothelin (ET), somatostatin (SS), acetylcholinesterase (AChE), substance P (SP) and vasoactive intestinal peptide (VIP) levels in serum were monitored to evaluate the preventive effects of LF-Zhao on constipation. Bisacodyl, a laxative drug, was used as a positive control. Times to the first black stool for normal (untreated), control (no lactic acid bacteria treatment but activated carbon treated), bisacodyl-treated and L. delbrueckii subsp. bulgaricus (LB), LF-Zhao (L) (low concentration of 1×10(8) CFU/mL)- and LF-Zhao (H) (high concentration of 1×10(9) CFU/mL)-treated mice induced by activated carbon were 90, 218, 117, 180, 169 and 156 min, respectively. Following the consumption of LB, LF-Zhao (L) and LF-Zhao (H) or the oral administration of bisacodyl, the gastrointestinal transit distances were reduced by 55.2%, 61.3%, 70.6% and 94.6%, respectively. The serum levels of MTL, Gas, ET, AChE, SP and VIP were significantly increased and the serum levels of SS were reduced in the mice treated with LF-Zhao compared with those in the control mice (p<0.05). These results demonstrated that lactic acid bacteria demonstrate preventive effects on mouse constipation and that LF-Zhao alleviated constipation symptoms better than LB. PMID:26052143

  18. Enhancement of host resistance against Listeria infection by Lactobacillus casei: Role of macrophages

    SciTech Connect

    Sato, K.

    1984-05-01

    Among the 10 species of the genus Lactobacillus, L. casei showed the strongest protective action against Listeria monocytogenes infection in mice. The activity of L. casei differed with regard to the dose of administration. The anti-L. monocytogenes resistance in mice intravenously administered 5.5 X 10(7), 2.8 X 10(8), or 1.1 X 10(9) L. casei cells was most manifest at ca. 2, 2 and 13, and 3 to 21 days after its administration, respectively. The growth of L. monocytogenes in the liver of mice injected with L. casei (10(7), 10(8), or 10(9) cells) 48 h after infection was suppressed, particularly when 10(8) or 10(9) L. casei cells were given 2 or 13 days before the induced infection, respectively. This suppression of L. monocytogenes growth was overcome by carrageenan treatment or X-ray irradiation. (/sup 3/H)thymidine incorporation into the liver DNA increased 13 days after administration of L. casei, and augmentation of (/sup 3/H)thymidine incorporation during 6 to 48 h after infection was dependent on the dose of L. casei. Peritoneal macrophage accumulation observed 1 to 5 days after intraperitoneal injection of UV-killed L. monocytogenes was markedly enhanced when the mice were treated with L. casei cells 13 days before macrophage elicitation. Therefore, the enhanced host resistance by L. casei to L. monocytogenes infection may be mediated by macrophages migrating from the blood stream to the reticuloendothelial system in response to L. casei injection before or after L. monocytogenes infection.

  19. Specific point mutations in Lactobacillus casei ATCC 27139 cause a phenotype switch from Lac- to Lac+.

    PubMed

    Tsai, Yu-Kuo; Chen, Hung-Wen; Lo, Ta-Chun; Lin, Thy-Hou

    2009-03-01

    Lactose metabolism is a changeable phenotype in strains of Lactobacillus casei. In this study, we found that L. casei ATCC 27139 was unable to utilize lactose. However, when exposed to lactose as the sole carbon source, spontaneous Lac(+) clones could be obtained. A gene cluster (lacTEGF-galKETRM) involved in the metabolism of lactose and galactose in L. casei ATCC 27139 (Lac(-)) and its Lac(+) revertant (designated strain R1) was sequenced and characterized. We found that only one nucleotide, located in the lacTEGF promoter (lacTp), of the two lac-gal gene clusters was different. The protein sequence identity between the lac-gal gene cluster and those reported previously for some L. casei (Lac(+)) strains was high; namely, 96-100 % identity was found and no premature stop codon was identified. A single point mutation located within the lacTp promoter region was also detected for each of the 41 other independently isolated Lac(+) revertants of L. casei ATCC 27139. The revertants could be divided into six classes based on the positions of the point mutations detected. Primer extension experiments conducted on transcription from lacTp revealed that the lacTp promoter of these six classes of Lac(+) revertants was functional, while that of L. casei ATCC 27139 was not. Northern blotting experiments further confirmed that the lacTEGF operon of strain R1 was induced by lactose but suppressed by glucose, whereas no blotting signal was ever detected for L. casei ATCC 27139. These results suggest that a single point mutation in the lacTp promoter was able to restore the transcription of a fully functional lacTEGF operon and cause a phenotype switch from Lac(-) to Lac(+) for L. casei ATCC 27139. PMID:19246746

  20. Inactivation of Lactobacillus leichmannii ribonucleotide reductase by F2CTP: covalent modification§

    PubMed Central

    Lohman, Gregory J. S.; Stubbe, JoAnne

    2010-01-01

    Ribonucleotide reductase (RNR) from Lactobacillus leichmannii, a 76 kDa monomer using adenosylcobalamin (AdoCbl) as a cofactor, catalyzes the conversion of nucleoside triphosphates to deoxynucleotides and is rapidly (<30 sec) inactivated by one equivalent (eq.) of 2?,2?-difluoro-2?-deoxycytidine 5?-triphosphate (F2CTP). [1?-3H] and [5-3H]-F2CTP were synthesized and used independently to inactivate RNR. Sephadex G-50 chromatography of the inactivation mixture revealed that 0.47 eq. of a sugar were covalently bound to RNR and that 0.71 eq. of cytosine were released. Alternatively, analysis of the inactivated RNR by SDS PAGE without boiling, resulted in 33% of RNR migrating as a 110 kDa protein. Inactivation of RNR with a mixture of [1?-3H]-F2CTP/[1?-2H]-F2CTP followed by reduction with NaBH4, alklyation with iodoacetamide, trypsin digestion, and HPLC separation of the resulting peptides, allowed isolation and identification by MALDI-TOF mass spectrometry (MS) of a [3H/2H]-peptide containing C731 and C736 from the C-terminus of RNR accounting for 10% of the labeled protein. The MS analysis also revealed that the two cysteines were cross-linked to a furanone-species derived from the sugar of F2CTP. Incubation of [1?-3H]-F2CTP with C119S-RNR resulted in 0.3 eq. of sugar covalently bound to the protein and incubation with NaBH4 subsequent to inactivation resulted in trapping of 2?-fluoro-2?-deoxycytidine. These studies and the ones in the accompanying manuscript allow proposal of a mechanism of inactivation of RNR by F2CTP involving multiple reaction pathways. The proposed mechanisms share many common features with F2CDP inactivation of the class I RNRs. PMID:20088569

  1. Predictive modelling of Lactobacillus casei KN291 survival in fermented soy beverage.

    PubMed

    Zieli?ska, Dorota; Dorota, Zieli?ska; Ko?o?yn-Krajewska, Danuta; Danuta, Ko?o?yn-Krajewska; Goryl, Antoni; Antoni, Goryl; Motyl, Ilona

    2014-02-01

    The aim of the study was to construct and verify predictive growth and survival models of a potentially probiotic bacteria in fermented soy beverage. The research material included natural soy beverage (Polgrunt, Poland) and the strain of lactic acid bacteria (LAB) - Lactobacillus casei KN291. To construct predictive models for the growth and survival of L. casei KN291 bacteria in the fermented soy beverage we design an experiment which allowed the collection of CFU data. Fermented soy beverage samples were stored at various temperature conditions (5, 10, 15, and 20°C) for 28 days. On the basis of obtained data concerning the survival of L. casei KN291 bacteria in soy beverage at different temperature and time conditions, two non-linear models (r(2)= 0.68-0.93) and two surface models (r(2)=0.76-0.79) were constructed; these models described the behaviour of the bacteria in the product to a satisfactory extent. Verification of the surface models was carried out utilizing the validation data - at 7°C during 28 days. It was found that applied models were well fitted and charged with small systematic errors, which is evidenced by accuracy factor - Af, bias factor - Bf and mean squared error - MSE. The constructed microbiological growth and survival models of L. casei KN291 in fermented soy beverage enable the estimation of products shelf life period, which in this case is defined by the requirement for the level of the bacteria to be above 10(6) CFU/cm(3). The constructed models may be useful as a tool for the manufacture of probiotic foods to estimate of their shelf life period. PMID:24500482

  2. The paratransgenic potential of Lactobacillus kunkeei in the honey bee Apis mellifera.

    PubMed

    Rangberg, A; Mathiesen, G; Amdam, G V; Diep, D B

    2015-01-01

    The honey bee (Apis mellifera) is a domestic insect of high value to human societies, as a crop pollinator in agriculture and a model animal in scientific research. The honey bee, however, has experienced massive mortality worldwide due to the phenomenon Colony Collapse Disorder (CCD), resulting in alarming prospects for crop failure in Europe and the USA. The reasons for CCD are complex and much debated, but several honey bee pathogens are believed to be involved. Paratransgenesis is a Trojan horse strategy, where endogenous microorganisms are used to express effector molecules that antagonise pathogen development. For use in honey bees, paratransgenesis must rely on a set of criteria that the candidate paratransgenic microorganism must fulfil in order to obtain a successful outcome: (1) the candidate must be genetically modifiable to express effector molecules; (2) the modified organism should have no adverse effects on honey bee health upon reintroduction; and (3) it must survive together with other non-pathogenic bee-associated microorganisms. Lactic acid bacteria (LAB) are common gut bacteria in vertebrates and invertebrates, and some have naturally beneficial properties in their host. In the present work we aimed to find a potential paratransgenic candidate within this bacterial group for use in honey bees. Among isolated LAB associated with bee gut microbiota, we found the fructophilic Lactobacillus kunkeei to be the most predominant species during foraging seasons. Four genetically different strains of L. kunkeei were selected for further assessment. We demonstrated (1) that L. kunkeei is transformable; (2) that the transformed cells had no obvious adverse effect on honey bee survival; and (3) that transformed cells survived well in the gut environment of bees upon reintroduction. Our study demonstrates that L. kunkeei fulfils the three criteria for paratransgenesis and can be a suitable candidate for further research on this strategy in honey bees. PMID:25609652

  3. Hypolipidemic effects and safety of Lactobacillus reuteri 263 in a hamster model of hyperlipidemia.

    PubMed

    Huang, Wen-Ching; Chen, Yi-Ming; Kan, Nai-Wen; Ho, Chun-Sheng; Wei, Li; Chan, Ching-Hung; Huang, Hui-Yu; Huang, Chi-Chang

    2015-05-01

    We aimed to verify the beneficial effects of probiotic strain Lactobacillus reuteri 263 (Lr263) on hypolipidemic action in hamsters with hyperlipidemia induced by a 0.2% cholesterol and 10% lard diet (i.e., high-cholesterol diet (HCD)). Male Golden Syrian hamsters were randomly divided into two groups: normal (n = 8), standard diet (control), and experimental (n = 32), a HCD. After a two-week induction followed by a six-week supplementation with Lr263, the 32 hyperlipidemic hamsters were divided into four groups (n = 8 per group) to receive vehicle or Lr263 by oral gavage at 2.1, 4.2, or 10.5 × 10(9) cells/kg/day for 6 weeks, designated the HCD, 1X, 2X and 5X groups, respectively. The efficacy and safety of Lr263 supplementation were evaluated by lipid profiles of serum, liver and feces and by clinical biochemistry and histopathology. HCD significantly increased serum levels of total cholesterol (TC), triacylglycerol (TG) cholesterol, high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C), LDL-C/HDL-C ratio, hepatic and fetal TC and TG levels, and degree of fatty liver as compared with controls. Lr263 supplementation dose dependently increased serum HDL-C level and decreased serum TC, TG, LDL-C levels, LDL-C/HDL-C ratio, hepatic TC and TG levels, and fecal TG level. In addition, Lr263 supplementation had few subchronic toxic effects. Lr263 could be a potential agent with a hypolipidemic pharmacological effect. PMID:25988768

  4. Comparative Genomic and Functional Analysis of 100 Lactobacillus rhamnosus Strains and Their Comparison with Strain GG

    PubMed Central

    Pietilä, Taija E.; Järvinen, Hanna M.; Messing, Marcel; Randazzo, Cinzia L.; Paulin, Lars; Laine, Pia; Ritari, Jarmo; Caggia, Cinzia; Lähteinen, Tanja; Brouns, Stan J. J.; Satokari, Reetta; von Ossowski, Ingemar; Reunanen, Justus; Palva, Airi; de Vos, Willem M.

    2013-01-01

    Lactobacillus rhamnosus is a lactic acid bacterium that is found in a large variety of ecological habitats, including artisanal and industrial dairy products, the oral cavity, intestinal tract or vagina. To gain insights into the genetic complexity and ecological versatility of the species L. rhamnosus, we examined the genomes and phenotypes of 100 L. rhamnosus strains isolated from diverse sources. The genomes of 100 L. rhamnosus strains were mapped onto the L. rhamnosus GG reference genome. These strains were phenotypically characterized for a wide range of metabolic, antagonistic, signalling and functional properties. Phylogenomic analysis showed multiple groupings of the species that could partly be associated with their ecological niches. We identified 17 highly variable regions that encode functions related to lifestyle, i.e. carbohydrate transport and metabolism, production of mucus-binding pili, bile salt resistance, prophages and CRISPR adaptive immunity. Integration of the phenotypic and genomic data revealed that some L. rhamnosus strains possibly resided in multiple niches, illustrating the dynamics of bacterial habitats. The present study showed two distinctive geno-phenotypes in the L. rhamnosus species. The geno-phenotype A suggests an adaptation to stable nutrient-rich niches, i.e. milk-derivative products, reflected by the alteration or loss of biological functions associated with antimicrobial activity spectrum, stress resistance, adaptability and fitness to a distinctive range of habitats. In contrast, the geno-phenotype B displays adequate traits to a variable environment, such as the intestinal tract, in terms of nutrient resources, bacterial population density and host effects. PMID:23966868

  5. Effects of High Pressure on Inactivation Kinetics and Events Related to Proton Efflux in Lactobacillus plantarum

    PubMed Central

    Wouters, Patrick C.; Glaasker, Erwin; Smelt, Jan P. P. M.

    1998-01-01

    Knowledge of the mechanism of pressure-induced inactivation of microorganisms could be helpful in defining an effective, relatively mild pressure treatment as a means of decontamination, especially in combination with other physical treatments or antimicrobial agents. We have studied the effect of high pressure on Lactobacillus plantarum grown at pH 5.0 and 7.0. The classical inactivation kinetics were compared with a number of events related to the acid-base physiology of the cell, i.e., activity of F0F1 ATPase, intracellular pH, acid efflux, and intracellular ATP pool. Cells grown at pH 5.0 were more resistant to pressures of 250 MPa than were cells grown at pH 7.0. This difference in resistance may be explained by a higher F0F1 ATPase activity, better ability to maintain a ?pH, or a higher acid efflux of the cells grown at pH 5.0. After pressure treatment at 250 MPa, the F0F1 ATPase activity was decreased, the ability to maintain a ?pH was reduced, and the acid efflux was impaired. The ATP pool increased initially after mild pressure treatment and finally decreased after prolonged treatment. The observations on acid efflux and the ATP pool suggest that the glycolysis is affected by high pressure later than is the F0F1 ATPase activity. Although functions related to the membrane-bound ATPase activity were impaired, no morphological changes of the membrane could be observed. PMID:16349500

  6. Cold stress improves the ability of Lactobacillus plantarum L67 to survive freezing.

    PubMed

    Song, Sooyeon; Bae, Dong-Won; Lim, Kwangsei; Griffiths, Mansel W; Oh, Sejong

    2014-11-17

    The stress resistance of bacteria is affected by the physiological status of the bacterial cell and environmental factors such as pH, salts and temperature. In this study, we report on the stress response of Lactobacillus plantarum L67 after four consecutive freeze-thaw cycles. The cold stress response of the cold-shock protein genes (cspC, cspL and cspP) and ATPase activities were then evaluated. The cold stress was adjusted to 5 °C when the bacteria were growing at the mid-exponential phase. A comparative proteomic analysis was performed with two-dimensional gel electrophoresis (2D SDS-PAGE) and a matrix assisted laser desorption/ionization-mass spectrometer. Only 56% of the L. plantarum L67 cells without prior exposure to cold stress survived after four consecutive freeze-thaw cycles. However, 78% of the L. plantarum L67 cells that were treated with cold stress at 5 °C for 6 h survived after freeze-thaw conditions. After applying cold stress to the culture for 6h, the cells were then stored for 60 days at 5 °C, 25 °C and 35 °C separately. The cold-stressed culture of L. plantarum L67 showed an 8% higher viability than the control culture. After applying cold stress for 6h, the transcript levels of two genes (cspP and cspL) were up-regulated 1.4 (cspP) and 1.2 (cspL) times compared to the control. However, cspC was not up-regulated. A proteomic analysis showed that the proteins increased after a reduction of the incubation temperature to 5 °C. The importance of the expression of 13 other relevant proteins was also determined through the study. The exposure of L. plantarum cells to low temperatures aids their ability to survive through subsequent freeze-thaw processes and lyophilization. PMID:25261832

  7. Exopolysaccharide Produced by Lactobacillus Plantarum Induces Maturation of Dendritic Cells in BALB/c Mice

    PubMed Central

    Tang, Yanjun; Dong, Wei; Wan, Keyu; Zhang, Ligang; Li, Chun; Zhang, Lili; Liu, Ning

    2015-01-01

    Lactobacillus plantarum (L. plantarum) exopolysaccharide (EPS) is an important bioactive component in fermented functional foods. However, there is a lack of data concerning the effects of L. plantarum EPS on maturation of mouse dendritic cells (DCs). In this study, we purified L. plantarum EPS and examined its effects on cytokines production by dendritic cells in serum and intestinal fluid of BALB/c mice, then investigated its effects on phenotypic and functional maturation of mouse bone marrow-derived dendritic cells (BMDCs). Cytokines (nitric oxide, IL-12p70, IL-10 and RANTES) in serum and intestinal fluid were analyzed by enzyme linked immunosorbent assay (ELISA) after the mice received EPS for 2, 5 and 7 days, respectively. DCs derived from bone marrow of BALB/c mouse were treated with EPS, then the phenotypic maturation of BMDCs was analyzed using flow cytometer and the functional maturation of BMDCs was analyzed by ELISA, and, lastly, mixed lymphocyte proliferation was performed. We found the molecular weight of purified EPS was approximately 2.4×106 Da and it was composed of ribose, rhamnose, arabinose, xylose, mannose, glucose and galactose in a molar ratio of 2:1:1:10:4:205:215. We observed that L. plantarum EPS enriched production of nitric oxide, IL-12p70 and RANTES, and decreased the secretion of IL-10 in the serum or intestinal fluid as well as in the supernatant of DCs treated with the EPS. The EPS also up-regulated the expression of MHC II and CD86 on DCs surface and promoted T cells to proliferate in vitro. Our data provide direct evidence to suggest that L. plantarum EPS can effectively induce maturation of DCs in mice. PMID:26599612

  8. Isolation and characterization of Lactobacillus salivarius MTC 1026 as a potential probiotic.

    PubMed

    Tinrat, Sirikhwan; Saraya, Sumarn; Traidej Chomnawang, Mullika

    2011-01-01

    Researchers are becoming more interested in studying probiotics at present due to their benefit to human and animal health. In this study, newly isolated strains from human feces were evaluated for probiotic properties. A total of sixty isolated strains were collected from feces and six out of sixty isolated strains could inhibit the growth of Salmonella typhi and Salmonella typhimurium. The strain which gave the best inhibitory effect was selected for further characterization as a probiotic strain. The identification of this strain was analyzed on the basis of morphological and biochemical characteristics and 16S rDNA gene sequence. This strain was Gram-positive, rod shaped, and catalase and oxidase-negative, and produced acids from D-glucose, D-fructose, lactose, mannitol, sorbitol, inulin and starch. It could not hydrolyze esculin or red blood cells. Based on its 16S rDNA gene sequence, it was Lactobacillus salivarius, and so was called L. salivarius MTC 1026 in this study, and was closely related with L. salivarius DSPV 344T isolated from the calf gut. It was able to survive in gastric and small intestinal juices at pH 2.0 and 1.0% bile salt for several hours and also could grow at 45°C. Moreover, this strain showed inhibitory activity against a variety of food-borne pathogens. It was highly sensitive to piperacillin, chloramphenicl, ampicillin, erythromycin and ceftazidime. After plasmid curing, this strain was susceptible to gentamicin, amikacin, norfloxacin and ciprofloxacin. L. salivarius MTC 1026 could significantly inhibit the adhesion process of E. coli ATCC 25922 and S. typhimurium ATCC 13311 on Caco-2 monolayers in a competition assay and also reduced invasion of both pathogens (4-log cfu/ml) in an exclusion and displacement assay. Therefore, it was clearly demonstrated in this study that L. salivarius MTC 1026 has shown promising properties as a candidate for a potential probiotic for applications in humans and animals. PMID:22353742

  9. Survival of Lactobacillus casei strain Shirota in the intestines of healthy Chinese adults.

    PubMed

    Wang, Ran; Chen, Shanbin; Jin, Junhua; Ren, Fazheng; Li, Yang; Qiao, Zhenxing; Wang, Yue; Zhao, Liang

    2015-05-01

    Lactobacillus casei strain Shirota (LcS) is a widely used probiotic strain with health benefits. In this study, the survival of LcS in the intestines of healthy Chinese adults was assessed and the effects of LcS on stool consistency, stool SCFAs and intestinal microbiota evaluated. Subjects consumed 100?mL per day of a probiotic beverage containing 1.0?×?10(8) CFU/mL of LcS for 14 days. LcS were enumerated using a culture method and the colony identity confirmed by ELISA. Fourteen days after ingestion, the amount of LcS recovered from fecal samples was between 6.86?±?0.80 and 7.17?±?0.57 Log10 CFU/g of feces (mean?±?SD). The intestinal microbiotas were analyzed by denaturing gradient gel electrophoresis. Principal component analysis showed that consuming LcS significantly changed fecal microbiota profiles. According to redundancy analysis, the amounts of 25 bacterial strains were significantly correlated with LcS intake (P < 0.05), 11 of them positively and fourteen negatively. Concentrations of acetic acid and propionic acid in feces were significantly lower during the ingestion period than during the baseline period (P < 0.05). These results confirm that LcS can survive passage through the gastrointestinal tract of Chinese people; however, they were found to have little ability to persist once their consumption had ceased. Furthermore, consumption of probiotic beverages containing LcS can modulate the composition of the intestinal microbiota on a long-term basis, resulting in decreased concentrations of SCFAs in the gut. PMID:25707300

  10. Metabolic responses of Lactobacillus plantarum strains during fermentation and storage of vegetable and fruit juices.

    PubMed

    Filannino, P; Cardinali, G; Rizzello, C G; Buchin, S; De Angelis, M; Gobbetti, M; Di Cagno, R

    2014-04-01

    Strains of Lactobacillus plantarum were grown and stored in cherry (ChJ), pineapple (PJ), carrot (CJ), and tomato (TJ) juices to mimic the chemical composition of the respective matrices. Wheat flour hydrolysate (WFH), whey milk (W), and MRS broth were also used as representatives of other ecosystems. The growth rates and cell densities of L. plantarum strains during fermentation (24 h at 30°C) and storage (21 days at 4°C) differed only in part, being mainly influenced by the matrix. ChJ and PJ were the most stressful juices for growth and survival. Overall, the growth in juices was negatively correlated with the initial concentration of malic acid and carbohydrates. The consumption of malic acid was noticeable for all juices, but mainly during fermentation and storage of ChJ. Decreases of branched-chain amino acids (BCAA)-with the concomitant increase of their respective branched alcohols-and His and increases of Glu and gamma-aminobutyric acid (GABA) were the main traits of the catabolism of free amino acids (FAA), which were mainly evident under less acidic conditions (CJ and TJ). The increase of Tyr was found only during storage of ChJ. Some aldehydes (e.g., 3-methyl-butanal) were reduced to the corresponding alcohols (e.g., 3-methyl-1-butanol). After both fermentation and storage, acetic acid increased in all fermented juices, which implied the activation of the acetate kinase route. Diacetyl was the ketone found at the highest level, and butyric acid increased in almost all fermented juices. Data were processed through multidimensional statistical analyses. Except for CJ, the juices (mainly ChJ) seemed to induce specific metabolic traits, which differed in part among the strains. This study provided more in-depth knowledge on the metabolic mechanisms of growth and maintenance of L. plantarum in vegetable and fruit habitats, which also provided helpful information to select the most suitable starters for fermentation of targeted matrices. PMID:24487533

  11. Characterization of nitrite degradation by Lactobacillus casei subsp. rhamnosus LCR 6013.

    PubMed

    Liu, Dong-mei; Wang, Pan; Zhang, Xin-yue; Xu, Xi-lin; Wu, Hui; Li, Li

    2014-01-01

    Nitrites are potential carcinogens. Therefore, limiting nitrites in food is critically important for food safety. The nitrite degradation capacity of Lactobacillus casei subsp. rhamnosus LCR 6013 was investigated in pickle fermentation. After LCR 6013 fermentation for 120 h at 37°C, the nitrite concentration in the fermentation system was significantly lower than that in the control sample without the LCR 6013 strain. The effects of NaCl and Vc on nitrite degradation by LCR 6013 in the De Man, Rogosa and Sharpe (MRS) medium were also investigated. The highest nitrite degradations, 9.29 mg/L and 9.89 mg/L, were observed when NaCl and Vc concentrations were 0.75% and 0.02%, respectively in the MRS medium, which was significantly higher than the control group (p ? 0.01). Electron capture/gas chromatography and indophenol blue staining were used to study the nitrite degradation pathway of LCR 6013. The nitrite degradation products contained N2O, but no NH4(+). The LCR 6013 strain completely degraded all NaNO2 (50.00 mg/L) after 16 h of fermentation. The enzyme activity of NiR in the periplasmic space was 2.5 times of that in the cytoplasm. Our results demonstrated that L. casei subsp. rhamnosus LCR 6013 can effectively degrade nitrites in both the pickle fermentation system and in MRS medium by NiR. Nitrites are degraded by the LCR 6013 strain, likely via the nitrate respiration pathway (NO2(-)>NO->N2O->N2), rather than the aammonium formation pathway (dissimilatory nitrate reduction to ammonium, DNRA), because the degradation products contain N2O, but not NH4(+). PMID:24755671

  12. Purification and Characterization of a Feruloyl Esterase from the Intestinal Bacterium Lactobacillus acidophilus

    PubMed Central

    Wang, Xiaokun; Geng, Xin; Egashira, Yukari; Sanada, Hiroo

    2004-01-01

    Dietary ferulic acid (FA), a significant antioxidant substance, is currently the subject of extensive research. FA in cereals exists mainly as feruloylated sugar ester. To release FA from food matrices, it is necessary to cleave ester cross-linking by feruloyl esterase (FAE) (hydroxycinnamoyl esterase; EC 3.1.1.73). In the present study, the FAE from a human typical intestinal bacterium, Lactobacillus acidophilus, was isolated, purified, and characterized for the first time. The enzyme was purified in successive steps including hydrophobic interaction chromatography and anion-exchange chromatography. The purified FAE appeared as a single band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with an apparent molecular mass of 36 kDa. It has optimum pH and temperature characteristics (5.6 and 37°C, respectively). The metal ions Cu2+ and Fe3+ (at a concentration of 5 mmol liter?1) inhibited FAE activity by 97.25 and 94.80%, respectively. Under optimum pH and temperature with 5-O-feruloyl-l-arabinofuranose (FAA) as a substrate, the enzyme exhibited a Km of 0.0953 mmol liter?1 and a Vmax of 86.27 mmol liter?1 min?1 mg?1 of protein. Furthermore, the N-terminal amino acid sequence of the purified FAE was found to be A R V E K P R K V I L V G D G A V G S T. The FAE released FA from O-(5-O-feruloyl-?-l-arabinofuranosyl)-(1?3)-O-?-d-xylopyranosyl-(1?4)-d-xylopyranose (FAXX) and FAA obtained from refined corn bran. Moreover, it released two times more FA from FAXX in the presence of added xylanase. PMID:15066833

  13. An Inserted ?/? Subdomain Shapes the Catalytic Pocket of Lactobacillus johnsonii Cinnamoyl Esterase

    PubMed Central

    Vu, Clara; Xu, Xiaohui; Cui, Hong; Molloy, Sara; Savchenko, Alexei; Yakunin, Alexander; Gonzalez, Claudio F.

    2011-01-01

    Background Microbial enzymes produced in the gastrointestinal tract are primarily responsible for the release and biochemical transformation of absorbable bioactive monophenols. In the present work we described the crystal structure of LJ0536, a serine cinnamoyl esterase produced by the probiotic bacterium Lactobacillus johnsonii N6.2. Methodology/Principal Findings We crystallized LJ0536 in the apo form and in three substrate-bound complexes. The structure showed a canonical ?/? fold characteristic of esterases, and the enzyme is dimeric. Two classical serine esterase motifs (GlyXSerXGly) can be recognized from the amino acid sequence, and the structure revealed that the catalytic triad of the enzyme is formed by Ser106, His225, and Asp197, while the other motif is non-functional. In all substrate-bound complexes, the aromatic acyl group of the ester compound was bound in the deepest part of the catalytic pocket. The binding pocket also contained an unoccupied area that could accommodate larger ligands. The structure revealed a prominent inserted ?/? subdomain of 54 amino acids, from which multiple contacts to the aromatic acyl groups of the substrates are made. Inserts of this size are seen in other esterases, but the secondary structure topology of this subdomain of LJ0536 is unique to this enzyme and its closest homolog (Est1E) in the Protein Databank. Conclusions The binding mechanism characterized (involving the inserted ?/? subdomain) clearly differentiates LJ0536 from enzymes with similar activity of a fungal origin. The structural features herein described together with the activity profile of LJ0536 suggest that this enzyme should be clustered in a new group of bacterial cinnamoyl esterases. PMID:21876742

  14. Anti-Inflammatory Effects of Lactobacillus Rahmnosus and Bifidobacterium Breve on Cigarette Smoke Activated Human Macrophages

    PubMed Central

    Mortaz, Esmaeil; Adcock, Ian M.; Ricciardolo, Fabio L. M.; Varahram, Mohammad; Jamaati, Hamidreza; Velayati, Ali Akbar; Folkerts, Gert; Garssen, Johan

    2015-01-01

    Background Chronic obstructive pulmonary disease (COPD) is a major global health problem with cigarette smoke (CS) as the main risk factor for its development. Airway inflammation in COPD involves the increased expression of inflammatory mediators such as CXCL-8 and IL-1? which are important mediators for neutrophil recruitment. Macrophages are an important source of these mediators in COPD. Lactobacillus rhamnosus (L. rhamnosus) and Befidobacterium breve (B. breve) attenuate the development of ‘allergic asthma’ in animals but their effects in COPD are unknown. Objective To determine the anti-inflammatory effects of L. rhamnosus and B. breve on CS and Toll-like receptor (TLR) activation. Design We stimulated the human macrophage cell line THP-1 with CS extract in the presence and absence of L. rhamnosus and B. breve and measured the expression and release of inflammatory mediators by RT-qPCR and ELISA respectively. An activity assay and Western blotting were used to examine NF-?B activation. Results Both L. rhamnosus and B. breve were efficiently phagocytized by human macrophages. L. rhamnosus and B. breve significantly suppressed the ability of CS to induce the expression of IL-1?, IL-6, IL-10, IL-23, TNF?, CXCL-8 and HMGB1 release (all p<0.05) in human THP-1 macrophages. Similar suppression of TLR4- and TLR9-induced CXCL8 expression was also observed (p<0.05). The effect of L. rhamnosus and B. breve on inflammatory mediator release was associated with the suppression of CS-induced NF-?B activation (p<0.05). Conclusions This data indicate that these probiotics may be useful anti-inflammatory agents in CS-associated disease such as COPD. PMID:26317628

  15. Fermentation metabolites from Lactobacillus gasseri and Propionibacterium freudenreichii exert bacteriocidal effects in mice.

    PubMed

    Kato-Mori, Yuko; Orihashi, Takenori; Kanai, Yuta; Sato, Michiko; Sera, Kenji; Hagiwara, Katsuro

    2010-12-01

    Lactobacillus gasseri OLL 2716 promotes the elimination of Helicobacter pylori and is utilized in yogurts that are specifically labeled as health foods. On the other hand, milk whey fermented by Propionibacterium freudenreichii ET-3, which increases the numbers of Bifidobacterium, is effective for intestinal disorders. We previously demonstrated that oral administration of L. gasseri and P. freudenreichii fermentation metabolites (LP-FM) improved calf intestinal microflora and reduced the incidence of diarrhea. However, the detailed immunological mechanisms responsible for these effects remain to be fully understood. In this study, we investigated whether LP-FM stimulates the innate immune response and promotes the elimination of Listeria monocytogenes in mice. The C57BL/6 female mice that were treated with LP-FM or L. gasseri fermentation metabolites alone for 4 weeks had more peripheral white blood cells than the untreated control mice. In particular, LP-FM-treated mice had higher CD4- and CD8-positive T-cell counts. The levels of reactive oxygen and nitrogen species produced by peritoneal macrophages were also higher in LP-FM-treated mice. Furthermore, LP-FM-treated mice that were infected with L. monocytogenes exhibited significant enhancement of the elimination of Listeria from the spleen and the liver in comparison with untreated control mice infected with Listeria. The activation of innate immunity by LP-FM was increased by the combination of fermentation metabolites from P. freudenreichii. These results suggest that LP-FM, which contains metabolites from L. gasseri and P. freudenreichii, stimulates the function of the innate immune system, thereby significantly promoting the elimination of L. monocytogenes in mice. PMID:20954806

  16. Enhanced D-lactic acid production from renewable resources using engineered Lactobacillus plantarum.

    PubMed

    Zhang, Yixing; Vadlani, Praveen V; Kumar, Amit; Hardwidge, Philip R; Govind, Revathi; Tanaka, Tsutomu; Kondo, Akihiko

    2016-01-01

    D-lactic acid is used as a monomer in the production of poly-D-lactic acid (PDLA), which is used to form heat-resistant stereocomplex poly-lactic acid. To produce cost-effective D-lactic acid by using all sugars derived from biomass efficiently, xylose-assimilating genes encoding xylose isomerase and xylulokinase were cloned into an L-lactate-deficient strain, Lactobacillus plantarum. The resulting recombinant strain, namely L. plantarum NCIMB 8826 ?ldhL1-pLEM-xylAB, was able to produce D-lactic acid (at optical purity >99 %) from xylose at a yield of 0.53 g g(-1). Simultaneous utilization of glucose and xylose to produce D-lactic acid was also achieved by this strain, and 47.2 g L(-1) of D-lactic acid was produced from 37.5 g L(-1) glucose and 19.7 g L(-1) xylose. Corn stover and soybean meal extract (SBME) were evaluated as cost-effective medium components for D-lactic acid production. Optimization of medium composition using response surface methodology resulted in 30 % reduction in enzyme loading and 70 % reduction in peptone concentration. In addition, we successfully demonstrated D-lactic acid fermentation from corn stover and SBME in a fed-batch fermentation, which yielded 61.4 g L(-1) D-lactic acid with an overall yield of 0.77 g g(-1). All these approaches are geared to attaining high D-lactic acid production from biomass sugars to produce low-cost, highly thermostable biodegradable plastics. PMID:26433970

  17. Application of Lactobacillus acidophilus (LA 5) strain in fruit-based ice cream.

    PubMed

    Senanayake, Suraji A; Fernando, Sirimali; Bamunuarachchi, Arthur; Arsekularatne, Mariam

    2013-11-01

    A study was performed to apply a probiotic strain into fermented ice cream mix with suitable fruit bases to develop a value-added product with a substantial level of viable organisms for a sufficient shelf life. Pure direct vat strain culture of Lactobacillus acidophilus (LA 5) in freeze-dried form was inoculated into a mixture of ice cream, frozen, and the number of viable organisms during frozen storage for a period of time was enumerated, using turbidity measurements with a spectrophotometer. An ice cream sample prepared without the probiotic culture was compared with the test sample for quality, by testing the basic quality parameters for ice cream. Results show a reduction in the over run of the probiotic ice cream compared to the nonprobiotic ice cream. Significantly high level (P < 0.05) of total solids (42%), proteins (16.5%), and titratable acidity (2.2%) was observed in the test sample compared to the nonprobiotic ice cream. Significantly low pH level in the probiotic sample may be due to the lactic acid produced by the probiotic culture. No significant difference (P > 0.05) in the fat content in the two types of ice cream was observed. A significantly low level (P < 0.05) of melting in the probiotic one may have resulted from less over run, than the nonprobiotic sample. Rapid reduction in the viable cells during frozen storage occurred at -18°C and gradual adaptation occurred over the first 4 weeks. At the 10th week, 1.0 × 10(7) numbers of viable organisms were present in 1 g of the probiotic ice cream. Results show the presence of a sufficient number of viable organisms in the product for the 10-week period, which would be beneficial to consumers. PMID:24804052

  18. Acid Stress-Mediated Metabolic Shift in Lactobacillus sanfranciscensis LSCE1 ?

    PubMed Central

    Serrazanetti, Diana I.; Ndagijimana, Maurice; Sado-Kamdem, Sylvain L.; Corsetti, Aldo; Vogel, Rudi F.; Ehrmann, Matthias; Guerzoni, M. Elisabetta

    2011-01-01

    Lactobacillus sanfranciscensis LSCE1 was selected as a target organism originating from recurrently refreshed sourdough to study the metabolic rerouting associated with the acid stress exposure during sourdough fermentation. In particular, the acid stress induced a metabolic shift toward overproduction of 3-methylbutanoic and 2-methylbutanoic acids accompanied by reduced sugar consumption and primary carbohydrate metabolite production. The fate of labeled leucine, the role of different nutrients and precursors, and the expression of the genes involved in branched-chain amino acid (BCAA) catabolism were evaluated at pH 3.6 and 5.8. The novel application of the program XCMS to the solid-phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS) data allowed accurate separation and quantification of 2-methylbutanoic and 3-methylbutanoic acids, generally reported as a cumulative datum. The metabolites coming from BCAA catabolism increased up to seven times under acid stress. The gene expression analysis confirmed that some genes associated with BCAA catabolism were overexpressed under acid conditions. The experiment with labeled leucine showed that 2-methylbutanoic acid originated also from leucine. While the overproduction of 3-methylbutanoic acid under acid stress can be attributed to the need to maintain redox balance, the rationale for the production of 2-methylbutanoic acid from leucine can be found in a newly proposed biosynthesis pathway leading to 2-methylbutanoic acid and 3 mol of ATP per mol of leucine. Leucine catabolism to 3-methylbutanoic and 2-methylbutanoic acids suggests that the switch from sugar to amino acid catabolism supports growth in L. sanfranciscensis in restricted environments such as sourdough characterized by acid stress and recurrent carbon starvation. PMID:21335381

  19. A study on characterization of new bacteriocin produced from a novel strain of Lactobacillus spicheri G2 isolated from Gundruk- a fermented vegetable product of North East India: A novel bacteriocin production from Lactobacillus spicheri G2.

    PubMed

    Gautam, Neha; Sharma, Nivedita

    2015-09-01

    Bacteriocin producing Lactobacillus spicheri G2, isolated from Gundruk - a traditional fermented vegetable product of North East India. L. spicheri G2 identified by morphological, biochemical techniques followed by 16S rRNA gene technique. The 16Sr RNA sequence of bacteriocin producer is registered in NCBI under accession no. JX481912. The bacteriocin producing potential of L. spicheri is being reported for the first time in the present investigation. Bacteriocin of L. spicheri G2 showed strong antagonism against food spoiling and pathogenic bacteria viz. Listeria monocytogenes, Staphlococcus aureus, Clostridium perfringens, Streptococcus mutans, Lactobacillus plantarum, Leuconostoc mesenteroides and Bacillus cereus. Bacteriocin production of L. spicheri G2 was enhanced by optimization of production time, pH of medium and incubation temperature by following one variable at a time method. Maximum bacteriocin activity (2000 AU/ml) was recorded in MRS broth at 34 h with an initial pH of 4.0 after incubating at 35 °C. The bacteriocin was purified by single step gel exclusion chromatography. Molecular weight of this novel bacteriocin was determined by SDS PAGE which was found to be 43 kDa. Purified bacteriocin was found resistant to high temperature and varied pH range but sensitive to proteolytic enzymes like trypsin and proteinase k, the characters desirable for food preservation. PMID:26344995

  20. A new high phenyl lactic acid-yielding Lactobacillus plantarum IMAU10124 and a comparative analysis of lactate dehydrogenase gene.

    PubMed

    Zhang, Xiqing; Zhang, Shuli; Shi, Yan; Shen, Fadi; Wang, Haikuan

    2014-07-01

    Phenyl lactic acid (PLA) has been widely reported as a new natural antimicrobial compound. In this study, 120 Lactobacillus plantarum strains were demonstrated to produce PLA using high-performance liquid chromatography. Lactobacillus plantarum IMAU10124 was screened with a PLA yield of 0.229 g L(-1) . Compared with all previous reports, this is the highest PLA-producing lactic acid bacteria (LAB) when grown in MRS broth without any optimizing conditions. When 3.0 g L(-1) phenyl pyruvic acid (PPA) was added to the medium as substrate, PLA production reached 2.90 g L(-1) , with the highest 96.05% conversion rate. A lowest PLA-yielding L. plantarum IMAU40105 (0.043 g L(-1) ) was also screened. It was shown that the conversion from PPA to PLA by lactic dehydrogenase (LDH) is the key factor in the improvement of PLA production by LAB. Comparing the LDH gene of two strains, four amino acid mutation sites were found in this study in the LDH of L. plantarum IMAU10124. PMID:24861375

  1. Synbiotic impact of tagatose on viability of Lactobacillus rhamnosus strain GG mediated by the phosphotransferase system (PTS).

    PubMed

    Koh, Ji Hoon; Choi, Seung Hye; Park, Seung Won; Choi, Nag-Jin; Kim, Younghoon; Kim, Sae Hun

    2013-10-01

    Synbiotics, the combination of prebiotics and probiotics, has been shown to produce synergistic effects that promote gastrointestinal well-being of host. Tagatose is a low calorie food ingredient with putative health-promoting benefits. Herein, we investigated its synbiotic impact on the viability of Lactobacillus casei 01 and Lactobacillus rhamnosus strain GG and the potential mechanism involved. Tagatose, as a synbiotic substrate, enhanced the growth of L. casei 01 and L. rhamnosus strain GG compared to other prebiotics. Other gut-indigenous such as Clostridium spp. readily utilized fructooligosaccharide (FOS), the most widely used functional prebiotics, but not tagatose. Additionally, tagatose enhanced probiotic functions of L. casei 01 and L. rhamnosus strain GG by reinforcing their attachment on HT-29 intestine epithelial cells and enhancing their cholesterol-lowering activities. Whole transcriptome study and quantitative real-time polymerase chain reaction (qRT-PCR) test showed that the presence of tagatose in L. rhamnosus strain GG caused induction of a large number of genes associated with carbohydrate metabolism including the phosphotransferase system (PTS). Collectively, these results indicate the tagatose enhanced the growth of L. casei 01 and L. rhamnosus strain GG and their probiotic activities by activating tagatose-associated PTS networks. Importantly, this study highlights the potential application of tagatose and L. casei 01 and/or L. rhamnosus strain GG as a synbiotic partner in functional dairy foods (i.e. yogurt and cheese) and therapeutic dietary supplements. PMID:23764214

  2. Complete nucleotide sequence of the 16S rRNA from Lactobacillus paracasei HS-05 isolated from women's hands.

    PubMed

    Choi, Woon Yong; Lee, Hyeon Yong

    2015-12-01

    We determined the complete nucleotide sequence of the 16S rRNA from a new bacterium collected from the surfaces of women's hands. We also compared the presence of various bacteria based on the subjects' sex and age. The number of colonies isolated from the hand surface was larger for women than men, and the largest number of isolates was confirmed to be present for the women in their 30 s and men in their 40 s (147 and 34 isolates, respectively). The morphology of an isolated bacterial strain was found to be rod type, and the bacterium was identified as Lactobacillaceae species based on the GenBank database, through a phylogenetic analysis using the 16S rRNA sequence. Based on the results of a homology search, the isolated strain was 99 % identical to Lactobacillus paracasei, so it was designated Lactobacillus paracasei HS-05 and was registered in the Korea Culture Center of Microorganisms (KCCM) database as [KCCM11349P]. PMID:26660786

  3. Antibacterial activity of a cell wall hydrolase from Lactobacillus paracasei NRRL B-50314 produced by recombinant Bacillus megaterium.

    PubMed

    Liu, Siqing; Rich, Joseph O; Anderson, Amber

    2015-02-01

    The cell-free supernatant (CFS) from Lactobacillus paracasei NRRL B-50314 culture has been previously reported as containing antibacterial activity against a wide variety of Gram-positive bacteria. The CFS protein gel slice corresponding to antibacterial activities was subjected to trypsin digestion and ion trap MASS (Gel/LC-MS/MS) analysis. BlastP search of the resulted IQAVISIAEQQIGKP sequence led to a hypothetical cell-wall associated hydrolase (designated as CWH here) from Lactobacillus paracasei ATCC 25302. Further analyses of CWH revealed that the IQAVISIAEQQIGKP belongs to a highly conserved region of the NlpC/P60 superfamily. The L. paracasei NRRL B-50314 CWH gene, cloned in pStrepHIS1525CWH477, was introduced into Bacillus megaterium MS 941. The production of CWH477 protein was induced by xylose. The CWH477 protein was purified by using NiNTA column, and elution fraction E2 showed highest antibacterial activity. This study and bioinformatics analyses suggested that the antibacterial activity of CWH could originate from its cell wall degrading enzymatic function. PMID:25533632

  4. In vitro and in vivo examination of anticolonization of pathogens by Lactobacillus paracasei FJ861111.1.

    PubMed

    Deng, Kan; Chen, Tingtao; Wu, Qinglong; Xin, Hongbo; Wei, Qiang; Hu, Ping; Wang, Xiaolei; Wang, Xin; Wei, Hua; Shah, Nagendra P

    2015-10-01

    Very limited information exists on the exclusion of pathogens by probiotics in the gut of the host challenged with pathogens. In this study, we tested probiotic characteristics in vitro and anticolonization ability of Lactobacillus paracasei FJ861111.1 in mice infected with selected pathogenic microorganisms. The in vitro results indicated that L. paracasei FJ861111.1 had a high survival in acidic conditions at pH 2.5 and bile salt concentration at 0.3%, and strong inhibition ability against common pathogens including Shigella dysenteriae, Staphylococcus aureus, Cronobacter sakazakii, Escherichia coli, and Candida albicans. The cell adhesion assays showed that L. paracasei FJ861111.1 exhibited strong adherence to HT-29 cells and excluded the adhesion of selected food-borne pathogens to HT-29 cells. The in vivo results showed that fermented milk with L. paracasei and viili (a Nordic yogurt product) significantly improved the population of total bacteria and of Lactobacillus in the feces of mice, and significantly inhibited the colonization of C. albicans to the intestines of mice post-C. albicans infection. Thus, it appears that this strain could be used as a probiotic organism for manufacturing functional fermented milk. PMID:26254535

  5. Significant Reduction in Helicobacter pylori Load in Humans with Non-viable Lactobacillus reuteri DSM17648: A Pilot Study.

    PubMed

    Holz, Caterina; Busjahn, Andreas; Mehling, Heidrun; Arya, Stefanie; Boettner, Mewes; Habibi, Hajar; Lang, Christine

    2015-06-01

    Reducing the amount of Helicobacter pylori in the stomach by selective bacterial-bacterial cell interaction was sought as an effective and novel method for combating the stomach pathogen. Lactobacillus reuteri DSM17648 was identified as a highly specific binding antagonist to H. pylori among more than 700 wild-type strains of Lactobacillus species. Applying a stringent screening procedure, the strain DSM17648 was identified as selective binder to H. pylori cells under in vivo gastric conditions. The strain DSM17648 co-aggregates the pathogen in vivo and in vitro. The specific co-aggregation occurs between Lact. reuteri DSM17648 and different H. pylori strains and serotypes, as well as H. heilmannii, but not with Campylobacter jejuni or other commensal oral and intestinal bacteria. Lact. reuteri DSM17648 was shown in a proof-of-concept single-blinded, randomized, placebo-controlled pilot study to significantly reduce the load of H. pylori in healthy yet infected adults. Reducing the amount of H. pylori in the stomach by selective bacterial-bacterial cell interaction might be an effective and novel method for combating the stomach pathogen. Lact. reuteri DSM17648 might prove useful as an adhesion blocker in antibiotic-free H. pylori therapies. PMID:25481036

  6. Metabolism of lactic acid in fermented cucumbers by Lactobacillus buchneri and related species, potential spoilage organisms in reduced salt fermentations.

    PubMed

    Johanningsmeier, Suzanne D; McFeeters, Roger F

    2013-09-01

    Recent evidence suggests that Lactobacillus buchneri may play an important role in spoilage-associated secondary fermentation of cucumbers. Lactic acid degradation during fermented cucumber spoilage is influenced by sodium chloride (NaCl) concentration, pH, and presence of oxygen. Objectives were to evaluate these factors on lactic acid utilization by L. buchneri, and to compare the biochemical changes to those which occur during fermented cucumber spoilage. Effects of NaCl (0, 2, 4, and 6% w/w), pH (3.8 vs 5.0), and aerobic environment were investigated using fermented cucumber media (FC) inoculated with spoilage microorganisms. At pH 3.8, L. buchneri degraded lactic acid in all NaCl concentrations. The highest rate of lactic acid utilization occurred in FC with 2% NaCl (P < 0.05). Lactic acid utilization was nearly identical under aerobic and anaerobic conditions, indicating that oxygen does not influence lactate metabolism by L. buchneri. Lactic acid utilization was accompanied by increases in acetic acid and 1,2-propanediol, and Lactobacillus rapi was able to convert 1,2-propanediol to propionic acid and propanol. L. buchneri initiated spoilage in a wide range of environmental conditions that may be present in commercial cucumber fermentations, and L. rapi may act syntrophically with L. buchneri to produce the commonly observed spoilage metabolites. PMID:23664264

  7. Lactobacillus acidophilus ATCC 4356 prevents atherosclerosis via inhibition of intestinal cholesterol absorption in apolipoprotein E-knockout mice.

    PubMed

    Huang, Ying; Wang, Jinfeng; Quan, Guihua; Wang, Xiaojun; Yang, Longfei; Zhong, Lili

    2014-12-01

    The objective of this study was to investigate the effect of Lactobacillus acidophilus ATCC 4356 on the development of atherosclerosis in apolipoprotein E-knockout (ApoE(-/-)) mice. Eight-week-old ApoE(-/-) mice were fed a Western diet with or without L. acidophilus ATCC 4356 daily for 16 weeks. L. acidophilus ATCC 4356 protected ApoE(-/-) mice from atherosclerosis by reducing their plasma cholesterol levels from 923 ± 44 to 581 ± 18 mg/dl, likely via a marked decrease in cholesterol absorption caused by modulation of Niemann-Pick C1-like 1 (NPC1L1). In addition, suppression of cholesterol absorption induced reverse cholesterol transport (RCT) in macrophages through the peroxisome proliferator-activated receptor/liver X receptor (PPAR/LXR) pathway. Fecal lactobacillus and bifidobacterium counts were significantly (P < 0.05) higher in the L. acidophilus ATCC 4356 treatment groups than in the control groups. Furthermore, L. acidophilus ATCC 4356 was detected in the rat small intestine, colon, and feces during the feeding trial. The bacterial levels remained high even after the administration of lactic acid bacteria had been stopped for 2 weeks. These results suggest that administration of L. acidophilus ATCC 4356 can protect against atherosclerosis through the inhibition of intestinal cholesterol absorption. Therefore, L. acidophilus ATCC 4356 may be a potential therapeutic material for preventing the progression of atherosclerosis. PMID:25261526

  8. The use of date waste for lactic acid production by a fed-batch culture using Lactobacillus casei subsp. rhamnosus

    PubMed Central

    Nancib, Aicha; Nancib, Nabil; Boubendir, Abdelhafid; Boudrant, Joseph

    2015-01-01

    The production of lactic acid from date juice by Lactobacillus caseisubsp. rhamnosus in batch and fed-batch cultures has been investigated. The fed-batch culture system gave better results for lactic acid production and volumetric productivity. The aim of this work is to determine the effects of the feeding rate and the concentration of the feeding medium containing date juice glucose on the cell growth, the consumption of glucose and the lactic acid production by Lactobacillus casei subsp. rhamnosus in fed-batch cultures. For this study, two concentrations of the feeding medium (62 and 100 g/L of date juice glucose) were tested at different feeding rates (18, 22, 33, 75 and 150 mL/h). The highest volumetric productivity (1.3 g/L.h) and lactic acid yield (1.7 g/g) were obtained at a feeding rate of 33 mL/h and a date juice glucose concentration of 62 g/L in the feeding medium. As a result, most of the date juice glucose was completely utilised (residual glucose 1 g/L), and a maximum lactic acid production level (89.2 g/L) was obtained. PMID:26413076

  9. The Highly Autoaggregative and Adhesive Phenotype of the Vaginal Lactobacillus plantarum Strain CMPG5300 Is Sortase Dependent

    PubMed Central

    Malik, Shweta; Petrova, Mariya I.; Claes, Ingmar J. J.; Verhoeven, Tine L. A.; Busschaert, Pieter; Vaneechoutte, Mario; Lievens, Bart; Lambrichts, Ivo; Siezen, Roland J.; Balzarini, Jan; Vanderleyden, Jos

    2013-01-01

    Lactobacilli are important for the maintenance of a healthy ecosystem in the human vagina. Various mechanisms are postulated but so far are poorly substantiated by molecular studies, such as mutant analysis. Bacterial autoaggregation is an interesting phenomenon that can promote adhesion to host cells and displacement of pathogens. In this study, we report on the identification of a human vaginal isolate, Lactobacillus plantarum strain CMPG5300, which shows high autoaggregative and adhesive capacity. To investigate the importance of sortase-dependent proteins (SDPs) in these phenotypes, a gene deletion mutant was constructed for srtA, the gene encoding the housekeeping sortase that covalently anchors these SDPs to the cell surface. This mutant lost the capacity to autoaggregate, showed a decrease in adhesion to vaginal epithelial cells, and lost biofilm-forming capacity under the conditions tested. These results indicate that the housekeeping sortase SrtA of CMPG5300 is a key determinant of the peculiar surface properties of this vaginal Lactobacillus strain. PMID:23709503

  10. Characterization and heterologous expression of a class IIa bacteriocin, plantaricin 423 from Lactobacillus plantarum 423, in Saccharomyces cerevisiae.

    PubMed

    Van Reenen, C A; Chikindas, M L; Van Zyl, W H; Dicks, L M T

    2003-02-25

    Lactobacillus plantarum 423 produces a small heat-stable antimicrobial protein designated plantaricin 423. This protein is bactericidal for many Gram-positive foodborne pathogens and spoilage bacteria, including Listeria spp., Staphylococcus spp., Pediococcus spp., Lactobacillus spp., etc. The DNA sequence of the plantaricin 423-encoding region on plasmid pPLA4 revealed a four open reading frame (ORF) operon structure similar to pediocin PA-1/AcH from Pediococcus acidilactici and coagulin from Bacillus coagulans I(4). The first ORF, plaA, encodes a 56-amino acid prepeptide consisting of a 37-amino acid mature molecule, with a 19-amino acid N-terminal leader peptide. The second ORF, plaB, encodes a putative immunity protein with protein sequence similarities to several bacteriocin immunity proteins. The plaC and plaD genes are virtually identical to pedC and pedD of the pediocin PA-1 operon, as well as coaC and coaD of the coagulin operon. Plantaricin 423 was cloned on a shuttle vector under the control of a yeast promoter and heterologously produced in Saccharomyces cerevisiae. PMID:12423916

  11. Overexpression and optimization of glutamate decarboxylase in Lactobacillus plantarum Taj-Apis362 for high gamma-aminobutyric acid production

    PubMed Central

    Tajabadi, Naser; Baradaran, Ali; Ebrahimpour, Afshin; Rahim, Raha A; Bakar, Fatimah A; Manap, Mohd Yazid A; Mohammed, Abdulkarim S; Saari, Nazamid

    2015-01-01

    Gamma-aminobutyric acid (GABA) is an important bioactive compound biosynthesized by microorganisms through decarboxylation of glutamate by glutamate decarboxylase (GAD). In this study, a full-length GAD gene was obtained by cloning the template deoxyribonucleic acid to pTZ57R/T vector. The open reading frame of the GAD gene showed the cloned gene was composed of 1410 nucleotides and encoded a 469 amino acids protein. To improve the GABA-production, the GAD gene was cloned into pMG36e-LbGAD, and then expressed in Lactobacillus plantarum?Taj-Apis362 cells. The overexpression was confirmed by SDS-PAGE and GAD activity, showing a 53?KDa protein with the enzyme activity increased by sevenfold compared with the original GAD activity. The optimal fermentation conditions for GABA production established using response surface methodology were at glutamic acid concentration of 497.973?mM, temperature 36°C, pH 5.31 and time 60?h. Under the conditions, maximum GABA concentration obtained (11.09?mM) was comparable with the predicted value by the model at 11.23?mM. To our knowledge, this is the first report of successful cloning (clone-back) and overexpression of the LbGAD gene from L.?plantarum to L.?plantarum cells. The recombinant Lactobacillus could be used as a starter culture for direct incorporation into a food system during fermentation for production of GABA-rich products. PMID:25757029

  12. Lactobacillus acidophilus ATCC 4356 Prevents Atherosclerosis via Inhibition of Intestinal Cholesterol Absorption in Apolipoprotein E-Knockout Mice

    PubMed Central

    Wang, Jinfeng; Quan, Guihua; Wang, Xiaojun; Yang, Longfei; Zhong, Lili

    2014-01-01

    The objective of this study was to investigate the effect of Lactobacillus acidophilus ATCC 4356 on the development of atherosclerosis in apolipoprotein E-knockout (ApoE?/?) mice. Eight-week-old ApoE?/? mice were fed a Western diet with or without L. acidophilus ATCC 4356 daily for 16 weeks. L. acidophilus ATCC 4356 protected ApoE?/? mice from atherosclerosis by reducing their plasma cholesterol levels from 923 ± 44 to 581 ± 18 mg/dl, likely via a marked decrease in cholesterol absorption caused by modulation of Niemann-Pick C1-like 1 (NPC1L1). In addition, suppression of cholesterol absorption induced reverse cholesterol transport (RCT) in macrophages through the peroxisome proliferator-activated receptor/liver X receptor (PPAR/LXR) pathway. Fecal lactobacillus and bifidobacterium counts were significantly (P < 0.05) higher in the L. acidophilus ATCC 4356 treatment groups than in the control groups. Furthermore, L. acidophilus ATCC 4356 was detected in the rat small intestine, colon, and feces during the feeding trial. The bacterial levels remained high even after the administration of lactic acid bacteria had been stopped for 2 weeks. These results suggest that administration of L. acidophilus ATCC 4356 can protect against atherosclerosis through the inhibition of intestinal cholesterol absorption. Therefore, L. acidophilus ATCC 4356 may be a potential therapeutic material for preventing the progression of atherosclerosis. PMID:25261526

  13. Purification and Molecular Characterization of the Novel Highly Potent Bacteriocin TSU4 Produced by Lactobacillus animalis TSU4.

    PubMed

    Sahoo, Tapasa Kumar; Jena, Prasant Kumar; Patel, Amiya Kumar; Seshadri, Sriram

    2015-09-01

    Bacterial infections causing fish diseases and spoilage during fish food processing and storage are major concerns in aquaculture. Use of bacteriocins has recently been considered as an effective strategy for prevention of bacterial infections. A novel bacteriocin produced by Catla catla gut isolates, Lactobacillus animalis TSU4, designated as bacteriocin TSU4 was purified to homogeneity by a three-step protocol. The molecular mass of bacteriocin TSU4 was 4117 Da determined by Q-TOF LC/MS analysis. Its isoelectric point was ~9. Secondary conformation obtained by circular dichroism spectroscopy showed molecular conformation with significant proportions of the structure in ?-helix (23.7 %) and ?-sheets (17.1 %). N-terminal sequencing was carried out by the Edman degradation method; partial sequence identified was NH2-SMSGFSKPHD. Bacteriocin TSU4 exhibited a wide range of antimicrobial activity, pH and thermal stability. It showed a bacteriocidal mode of action against the indicator strain Aeromonas hydrophila MTCC 646. Bacteriocin TSU4 is the first reported bacteriocin produced by fish isolate Lactobacillus animalis. The characterization of bacteriocin TSU4 suggested that it is a novel bacteriocin with potential value against infections of bacteria such as A. hydrophila MTCC 646 and Pseudomonas aeruginosa MTCC 1688 and application to prevent spoilage during food preservation. PMID:26245257

  14. Effect of Lactobacillus salivarius on Th1/Th2 cytokines and the number of spleen CD4+ CD25+ Foxp3+ Treg in asthma Balb/c mouse

    PubMed Central

    Yun, Xiang; Shang, Yunxiao; Li, Miao

    2015-01-01

    Background: Bronchial asthma is a chronic airway inflammatory disease that involves T lymphocytes. Methods: In order to explore the effect of Lactobacillus salivarius on Th1/Th2 cytokines and the number of spleen CD4+ CD25+ Foxp3+ Treg in asthma Balb/c mouse, we constructed acute asthma model with ovalbumin to observe the mouse behavior change in Balb/c mice. The expression of GATA-3 mRNA and T-bet mRNA was measured by real-time PCR. The proportion of CD4+ CD25+ Foxp3+ Treg/CD4+ was determined by flow cytometry. Results: The results demonstrated that oral gavage with Lactobacillus salivarius before sensitization could alleviate the clinical symptoms, airway hyper-reactivity and airway inflammation in asthma mouse to some extent; Lactobacillus salivarius may improve the imbalance of Th1/Th2 in asthma mouse through increasing the expression of T-bet mRNA at the transcriptional level and inhibiting the expression of GATA-3 mRNA simultaneously. Conclusion: CD4+ CD25+ Foxp3+ Treg cells may be involved in the pathogenesis of bronchial asthma, and may be the upstream regulatory mechanism of the improvement of Th1/Th2 imbalance by Lactobacillus salivarius. PMID:26339333

  15. Draft Genome Sequence of d-Branched-Chain Amino Acid Producer Lactobacillus otakiensis JCM 15040T, Isolated from a Traditional Japanese Pickle

    PubMed Central

    Mori, Kazuki; Mutaguchi, Yuta; Tashiro, Kosuke; Fujino, Yasuhiro; Ohmori, Taketo; Kuhara, Satoru; Ohshima, Toshihisa

    2013-01-01

    Lactobacillus otakiensis strain JCM 15040T was isolated from an unsalted pickling solution used in the production of sunki, a traditional Japanese pickle. Here, we prepared a draft genome sequence for this strain consisting of 40 contigs containing a total of 2,347,132 bp, 2,310 predicted coding sequences, and a G+C content of 42.4%. PMID:23929467

  16. Draft Genome Sequence of D-Branched-Chain Amino Acid Producer Lactobacillus otakiensis JCM 15040T, Isolated from a Traditional Japanese Pickle.

    PubMed

    Doi, Katsumi; Mori, Kazuki; Mutaguchi, Yuta; Tashiro, Kosuke; Fujino, Yasuhiro; Ohmori, Taketo; Kuhara, Satoru; Ohshima, Toshihisa

    2013-01-01

    Lactobacillus otakiensis strain JCM 15040(T) was isolated from an unsalted pickling solution used in the production of sunki, a traditional Japanese pickle. Here, we prepared a draft genome sequence for this strain consisting of 40 contigs containing a total of 2,347,132 bp, 2,310 predicted coding sequences, and a G+C content of 42.4%. PMID:23929467

  17. Anti-Infective Activities of Lactobacillus Strains in the Human Intestinal Microbiota: from Probiotics to Gastrointestinal Anti-Infectious Biotherapeutic Agents

    PubMed Central

    Liévin-Le Moal, Vanessa

    2014-01-01

    SUMMARY A vast and diverse array of microbial species displaying great phylogenic, genomic, and metabolic diversity have colonized the gastrointestinal tract. Resident microbes play a beneficial role by regulating the intestinal immune system, stimulating the maturation of host tissues, and playing a variety of roles in nutrition and in host resistance to gastric and enteric bacterial pathogens. The mechanisms by which the resident microbial species combat gastrointestinal pathogens are complex and include competitive metabolic interactions and the production of antimicrobial molecules. The human intestinal microbiota is a source from which Lactobacillus probiotic strains have often been isolated. Only six probiotic Lactobacillus strains isolated from human intestinal microbiota, i.e., L. rhamnosus GG, L. casei Shirota YIT9029, L. casei DN-114 001, L. johnsonii NCC 533, L. acidophilus LB, and L. reuteri DSM 17938, have been well characterized with regard to their potential antimicrobial effects against the major gastric and enteric bacterial pathogens and rotavirus. In this review, we describe the current knowledge concerning the experimental antibacterial activities, including antibiotic-like and cell-regulating activities, and therapeutic effects demonstrated in well-conducted, placebo-controlled, randomized clinical trials of these probiotic Lactobacillus strains. What is known about the antimicrobial activities supported by the molecules secreted by such probiotic Lactobacillus strains suggests that they constitute a promising new source for the development of innovative anti-infectious agents that act luminally and intracellularly in the gastrointestinal tract. PMID:24696432

  18. Probabilistic models to describe the effect of NaNO2 in combination with NaCl on the growth inhibition of Lactobacillus in frankfurters.

    PubMed

    Lee, Soomin; Lee, Heeyoung; Kim, Sejeong; Lee, Jeeyeon; Ha, Jimyeong; Gwak, Eunji; Oh, Mi-Hwa; Park, Beom-Young; Kim, Jin-Seok; Choi, Kyoung-Hee; Yoon, Yohan

    2015-12-01

    Probabilistic models were developed to describe the antimicrobial effect of NaNO2 (0-210ppm) in combination with NaCl (0-1.75%) on Lactobacillus growth under aerobic and anaerobic conditions. Growth (1) or no growth (0) was assessed every 24h as turbid or not turbid, respectively. The growth response data were analyzed by logistic regression to select significant variables (P<0.05) for Lactobacillus growth inhibition, and these variables were used to generate a probabilistic model. The model was then validated with observed data from frankfurters (a model system). NaNO2 and NaCl inhibited (P<0.05) Lactobacillus growth at all temperatures under aerobic and anaerobic conditions, and the antimicrobial effect of NaNO2 increased as the NaCl concentration increased. Validation showed that the performance of the developed model was appropriate. These results indicate that the models developed in this study should be useful for describing the antimicrobial effect of NaNO2 in combination with NaCl on Lactobacillus. PMID:26410421

  19. IMMUNOMODULATORY EFFECTS OF VITAMIN A DEFICIENCY AND LACTOBACILLUS-BASED PROBIOTIC ON HOST IMMUNE RESPONSES IN EIMERIA ACERVULINA-INFECTED BROILERS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Previously we have shown in separate experiments that resistance in broilers to Eimeria acervulina (EA) was reduced by vitamin A deficiency and enhanced by a probiotic (Lactobacillus-based, Primalac®). In the present 2 x 2 factorial study, a broiler starter ration was amended for vitamin A (control...

  20. A lactobacillus rhamnosus GG-derived soluble protein, p40, stimulates ligand release from intestinal epithelial cells to transactivate epidermal growth factor receptor

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Protein p40, a Lactobacillus rhamnosus GG (LGG)-derived soluble protein, ameliorates intestinal injury and colitis, reduces apoptosis and preserves barrier function by activation of EGF receptor (EGFR) in intestinal epithelial cells. The aim of this study was to determine the mechanisms by which p40...