Sample records for lactobacillus acidophilus ufv-h2b20

  1. Genes involved in lactose catabolism and organic acid production during growth of Lactobacillus delbrueckii UFV H2b20 in skimmed milk.

    PubMed

    Do Carmo, A P; De Oliveira, M N V; Da Silva, D F; Castro, S B; Borges, A C; De Carvalho, A F; De Moraes, C A

    2012-03-01

    There are three main reasons for using lactic acid bacteria (LAB) as starter cultures in industrial food fermentation processes: food preservation due to lactic acid production; flavour formation due to a range of organic molecules derived from sugar, lipid and protein catabolism; and probiotic properties attributed to some strains of LAB, mainly of lactobacilli. The aim of this study was to identify some genes involved in lactose metabolism of the probiotic Lactobacillus delbrueckii UFV H2b20, and analyse its organic acid production during growth in skimmed milk. The following genes were identified, encoding the respective enzymes: ldh - lactate dehydrogenase, adhE - Ldb1707 acetaldehyde dehydrogenase, and ccpA-pepR1 - catabolite control protein A. It was observed that L. delbrueckii UFV H2b20 cultivated in different media has the unexpected ability to catabolyse galactose, and to produce high amounts of succinic acid, which was absent in the beginning, raising doubts about the subspecies in question. The phylogenetic analyses showed that this strain can be compared physiologically to L. delbrueckii subsp. bulgaricus and L. delbrueckii subsp. lactis, which are able to degrade lactose and can grow in milk. L. delbrueckii UFV H2b20 sequences have grouped with L. delbrueckii subsp. bulgaricus ATCC 11842 and L. delbrueckii subsp. bulgaricus ATCC BAA-365, strengthening the classification of this probiotic strain in the NCFM group proposed by a previous study. Additionally, L. delbrueckii UFV H2b20 presented an evolutionary pattern closer to that of probiotic Lactobacillus acidophilus NCFM, corroborating the suggestion that this strain might be considered as a new and unusual subspecies among L. delbrueckii subspecies, the first one identified as a probiotic. In addition, its unusual ability to metabolise galactose, which was significantly consumed in the fermentation medium, might be exploited to produce low-browning probiotic Mozzarella cheeses, a desirable property

  2. Genes involved in protein metabolism of the probiotic lactic acid bacterium Lactobacillus delbrueckii UFV H2b20.

    PubMed

    Do Carmo, A P; da Silva, D F; De Oliveira, M N V; Borges, A C; De Carvalho, A F; De Moraes, C A

    2011-09-01

    A basic requirement for the prediction of the potential use of lactic acid bacteria (LAB) in the dairy industry is the identification of specific genes involved in flavour-forming pathways. The probiotic Lactobacillus delbrueckii UFV H2b20 was submitted to a genetic characterisation and phylogenetic analysis of genes involved in protein catabolism. Eight genes belonging to this system were identified, which possess a closely phylogenetic relationship to NCFM strains representative, as it was demonstrated for oppC and oppBII, encoding oligopeptide transport system components. PepC, PepN, and PepX might be essential for growth of LAB, probiotic or not, since the correspondent genes are always present, including in L. delbrueckii UFV H2b20 genome. For pepX gene, a probable link between carbohydrate catabolism and PepX expression may exists, where it is regulated by PepR1/CcpA-like, a common feature between Lactobacillus strains and also in L. delbrueckii UFV H2b20. The well conserved evolutionary history of the ilvE gene is evidence that the pathways leading to branched-chain amino acid degradation, such as isoleucine and valine, are similar among L. delbrueckii subsp. bulgaricus strains and L. delbrueckii UFV H2b20. Thus, the involvement of succinate in flavour formation can be attributed to IlvE activity. The presence of aminopeptidase G in L. delbrueckii UFV H2b20 genome, which is absent in several strains, might improve the proteolytic activity and effectiveness. The nucleotide sequence encoding PepG revealed that it is a cysteine endopeptidase, belonging to Peptidase C1 superfamily; sequence analysis showed 99% identity with L. delbrueckii subsp. bulgaricus ATCC 11842 pepG, whereas protein sequence analysis revealed 100% similarity with PepG from the same organism. The present study proposes a schematic model to explain how the proteolytic system of the probiotic L. delbrueckii UFV H2b20 works, based on the components identified so far.

  3. Involvement of the ornithine decarboxylase gene in acid stress response in probiotic Lactobacillus delbrueckii UFV H2b20.

    PubMed

    Ferreira, A B; Oliveira, M N V de; Freitas, F S; Paiva, A D; Alfenas-Zerbini, P; Silva, D F da; Queiroz, M V de; Borges, A C; Moraes, C A de

    2015-01-01

    Amino acid decarboxylation is important for the maintenance of intracellular pH under acid stress. This study aims to carry out phylogenetic and expression analysis by real-time PCR of two genes that encode proteins involved in ornithine decarboxylation in Lactobacillus delbrueckii UFV H2b20 exposed to acid stress. Sequencing and phylogeny analysis of genes encoding ornithine decarboxylase and amino acid permease in L. delbrueckii UFV H2b20 showed their high sequence identity (99%) and grouping with those of L. delbrueckii subsp. bulgaricus ATCC 11842. Exposure of L. delbrueckii UFV H2b20 cells in MRS pH 3.5 for 30 and 60 min caused a significant increase in expression of the gene encoding ornithine decarboxylase (up to 8.1 times higher when compared to the control treatment). Increased expression of the ornithine decarboxylase gene demonstrates its involvement in acid stress response in L. delbrueckii UFV H2b20, evidencing that the protein encoded by that gene could be involved in intracellular pH regulation. The results obtained show ornithine decarboxylation as a possible mechanism of adaptation to an acidic environmental condition, a desirable and necessary characteristic for probiotic cultures and certainly important to the survival and persistence of the L. delbrueckii UFV H2b20 in the human gastrointestinal tract.

  4. Survival of Lactobacillus delbrueckii UFV H2b20 in fermented milk under simulated gastric and intestinal conditions.

    PubMed

    da Conceição, L L; Leandro, E S; Freitas, F S; de Oliveira, M N V; Ferreira-Machado, A B; Borges, A C; de Moraes, C A

    2013-09-01

    The survival of Lactobacillus delbrueckii UFV H2b20 was assessed in fermented milk, both during the storage period and after exposure to simulated gastric and intestinal juices, as well the detection of the gene fbpA involved in adherence to human gastrointestinal tract. L. delbrueckii UFV H2b20 remained stable and viable for 28 days under refrigerated storage conditions. After one day of storage, that strain exhibited a one-log population reduction following exposure in tandem to simulated gastric and intestinal juices. After 14 days of storage, a two-log reduction was observed following 90 min of exposure to the simulated gastric conditions. However, the strain did not survive following exposure to the simulated intestinal juice. The observed tolerance to storage conditions and resistance to the simulated gastric and intestinal conditions confirm the potential use of L. delbrueckii UFV H2b20 as a probiotic, which is further reinforced by the detection of fbpA in this strain.

  5. Increased expression of clp genes in Lactobacillus delbrueckii UFV H2b20 exposed to acid stress and bile salts.

    PubMed

    Ferreira, A B; De Oliveira, M N V; Freitas, F S; Alfenas-Zerbini, P; Da Silva, D F; De Queiroz, M V; Borges, A C; De Moraes, C A

    2013-12-01

    The ability to survive in harsh environments is an important criterion to select potential probiotics strains. The objective of this study was to identify and carry out phylogenetic and expression analysis by quantitative real-time PCR of the clpP, clpE, clpL and clpX genes in the probiotic strain Lactobacillus delbrueckii UFV H2b20 exposed to the conditions prevailing in the gastrointestinal tract (GIT). Phylogenetic trees reconstructed by Bayesian inference showed that the L. delbrueckii UFV H2b20 clpP, clpL and clpE genes and the ones from L. delbrueckii ATCC 11842 were grouped. The exposure of cells to MRS broth of pH 3.5 for 30 and 60 min resulted in an increased expression of the four genes. Exposure of the L. delbrueckii UFV H2b20 cells for 30 and 60 min to MRS broth containing 0.1% bile salts increased the expression of the clpP and clpE genes, while the expression level of the clpL and clpX genes increased only after 30 min of exposure. The involvement of the studied genes in the responses to acid stress and bile salts suggests a possible central role of these genes in the survival of L. delbrueckii UFV H2b20 during the passage through the GIT, a characteristic necessary for probiotic strains.

  6. Monoassociation with probiotic Lactobacillus delbrueckii UFV-H2b20 stimulates the immune system and protects germfree mice against Listeria monocytogenes infection.

    PubMed

    dos Santos, Liliane Martins; Santos, Mônica Morais; de Souza Silva, Humberto Pereira; Arantes, Rosa Maria Esteves; Nicoli, Jacques Robert; Vieira, Leda Quercia

    2011-02-01

    In the present study, we investigated the protective effects of Lactobacillus delbrueckii UFV-H2b20 on the resistance to Listeria monocytogenes infection in gnotobiotic mice. Germfree mice or monoassociated mice were infected with L. monocytogenes, and the microbiological and immunological responses were evaluated after 1, 3, and 5 days of infection. Monoassociation with L. delbrueckii was capable of protecting mice against death caused by L. monocytogenes and induced a faster clearance of the bacteria in the liver, spleen, and peritoneal cavity at days 1, 3, and 5 post-infection. Also, monoassociated mice displayed less liver injury than germfree mice. The production of TNF-α in the serum, peritoneal cavity, and gut was augmented in monoassociated mice. Likewise, the levels of IFN-γ found on supernatants of spleen cells cultures were higher after the monoassociation. In addition, increased production of nitric oxide in peritoneal cell cultures supernatants and in serum was observed in mice that received L. delbrueckii. The monoassociation with L. delbrueckii induced higher production of IL-10 in the mucosal immune system. We conclude that monoassociation with L. delbrueckii UFV-H2b20 protects mice from death caused by L. monocytogenes infection by favoring effector responses while preventing their immunopathological consequences.

  7. Lactobacillus plantarum L9 but not Lactobacillus acidophilus LA reduces tumour necrosis factor induced bacterial translocation in Caco-2 cells.

    PubMed

    Wang, B; Chen, J; Wang, S; Zhao, X; Lu, G; Tang, X

    2017-05-30

    Translocation of bacteria across the intestinal barrier is important in the pathogenesis of systemic sepsis and multiple organ dysfunction syndromes. Inflammatory cytokines increase paracellular permeability that allows increased luminal bacteria to translocate across mucosal epithelium and further deteriorate the gut barrier. In order to reduce this risk, the prophylactic use of probiotics has been recently addressed. In this paper, we investigate the protective role toward tumour necrosis factor (TNF)-α induced non-pathogenic Escherichia coli translocation across Caco-2 monolayers of Lactobacillus strains. According to our experimental data, Lactobacillus plantarum L9 and Lactobacillus acidophilus LA have good capacities to adhere to Caco-2 cells. Addition of L. plantarum L9 and L. acidophilus LA to the enterocyte monolayer surface result in significant inhibition of E. coli adhesion and cell internalisation. However, L. plantarum L9 and L. acidophilus LA did not inhibit the growth of the non-pathogenic E. coli B5 after 24 h incubation. Exposure to TNF-α for 6 h caused a dramatic increase in E. coli B5 translocation across Caco-2 cells, which was uncoupled from increases in paracellular permeability. Pretreatment with L. plantarum L9 prevent TNF-α induced transcellular bacterial translocation and IL-8 production in Caco-2 cells. L. plantarum L9 also did not affect the integrity of the monolayers, as indicated by lactate dehydrogenase release, horseradish peroxidase permeability, and transepithelial electrical resistance. L. plantarum L9 showed the potential to protect enterocytes from an acute inflammatory response and therefore could be good potential prophylactic agents in counteracting bacterial translocation.

  8. Lactobacillus acidophilus modulates the virulence of Clostridium difficile.

    PubMed

    Yun, B; Oh, S; Griffiths, M W

    2014-01-01

    Clostridium difficile is a spore-forming, toxin-producing, anaerobic bacterium that colonizes the human gastrointestinal tract. This pathogen causes antibiotic-associated diarrhea and colitis in animals and humans. Antibiotic-associated diseases may be treated with probiotics, and interest is increasing in such uses of probiotics. This study investigated the effect of Lactobacillus strains on the quorum-sensing signals and toxin production of C. difficile. In addition, an in vivo experiment was designed to assess whether Lactobacillus acidophilus GP1B is able to control C. difficile-associated disease. Autoinducer-2 activity was measured for C. difficile using the Vibrio harveyi coupled bioluminescent assay. Cell extract (10μg/mL) of L. acidophilus GP1B exhibited the highest inhibitory activity among 5 to 40μg/mL cell-extract concentrations. Real-time PCR data indicated decreased transcriptional levels in luxS, tcdA, tcdB, and txeR genes in the presence of 10μg/mL of cell extract of L. acidophilus GP1B. Survival rates at 5d for mice given the pathogen alone with L. acidophilus GP1B cell extract or L. acidophilus GP1B were 10, 70, and 80%, respectively. In addition, the lactic acid-produced L. acidophilus GP1B exhibits an inhibitory effect against the growth of C. difficile. Both the L. acidophilus GP1B and GP1B cell extract have significant antipathogenic effects on C. difficile. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  9. Effects of salt concentration and pH on structural and functional properties of Lactobacillus acidophilus: FT-IR spectroscopic analysis.

    PubMed

    Gandhi, Akanksha; Shah, Nagendra P

    2014-03-03

    The effects of sodium chloride concentration and varying pH levels on the structural and functional properties of Lactobacillus acidophilus were investigated. Reconstituted skim milk was inoculated with Lb. acidophilus at varying salt concentrations (0, 1, 2, 5 and 10% NaCl) and pH levels (4.0, 5.0 and 6.0) and ACE-inhibitory activity and proteolytic activity were determined and the viable cell count was enumerated after 24h of fermentation at 37 °C. The degree of proteolysis exhibited an increase with higher salt concentration at pH 5.0 and 6.0. ACE-inhibitory activity was found to be the highest at pH 5.0 at all salt concentrations. Fourier transform infrared spectroscopy results demonstrated significant changes occurring beyond 2% NaCl particularly at low pH (4.0). The findings revealed that significant changes occurred in amide I and amide III regions when Lb. acidophilus was subjected to varying salt concentrations. Copyright © 2014 Elsevier B.V. All rights reserved.

  10. Molecular cloning, expression and adhesion analysis of silent slpB of Lactobacillus acidophilus NCFM.

    PubMed

    Guo, Yuxing; Li, Xiangyue; Yang, Yao; Wu, Zhen; Zeng, Xiaoqun; Nadari, Fawze; Pan, Daodong

    2018-06-23

    The slpB gene of Lactobacillus acidophilus NCFM, which differs from the slpA gene and is silent under normal conditions, was successfully amplified and ligated to the corresponding available sites on a recombinant pET-28a vector. Then the pET-28a-slpB vector was transformed into Escherichia coli DH (DE3) and the fusion His-slpB protein was expressed by induction with 1 mM IPTG for 14 h at 37 °C. The resulting His-slpB protein (S B ) had a relative molecular weight of 48 kDa. It was purified using a Ni-NTA column and was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and western blot contrastive analysis. The slpA protein (S A ) from L. acidophilus NCFM was extracted and purified. It had a relative molecular weight of 46 kDa. Circular dichroism measurements suggested that the two S-layer proteins had a high β-sheet content and a low α-helix structure content. In an adhesion experiment, S A displayed higher adhesive capability towards Caco-2 cells than did S B . The results suggest that these two S-layer proteins could have biotechnological applications.

  11. Protection of Lactobacillus acidophilus NRRL-B 4495 under in vitro gastrointestinal conditions with whey protein/pullulan microcapsules.

    PubMed

    Çabuk, Burcu; Tellioğlu Harsa, Şebnem

    2015-12-01

    In this research, whey protein/pullulan (WP/pullulan) microcapsules were developed in order to assess its protective effect on the viability of Lactobacillus acidophilus NRRL-B 4495 under in vitro gastrointestinal conditions. Results demonstrated that WP/pullulan microencapsulated cells exhibited significantly (p ≤ 0.05) higher resistance to simulated gastric acid and bile salt. Pullulan incorporation into protein wall matrix resulted in improved survival as compared to free cells after 3 h incubation in simulated gastric solution. Moreover WP/pullulan microcapsules were found to release over 70% of encapsulated L. acidophilus NRRL-B 4495 cells within 1 h. The effect of encapsulation during refrigerated storage was also studied. Free bacteria exhibited 3.96 log reduction while, WP/pullulan encapsulated bacteria showed 1.64 log reduction after 4 weeks of storage. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  12. H2O2 Production in Species of the Lactobacillus acidophilus Group: a Central Role for a Novel NADH-Dependent Flavin Reductase

    PubMed Central

    Hertzberger, Rosanne; Arents, Jos; Dekker, Henk L.; Pridmore, R. David; Gysler, Christof; Kleerebezem, Michiel

    2014-01-01

    Hydrogen peroxide production is a well-known trait of many bacterial species associated with the human body. In the presence of oxygen, the probiotic lactic acid bacterium Lactobacillus johnsonii NCC 533 excretes up to 1 mM H2O2, inducing growth stagnation and cell death. Disruption of genes commonly assumed to be involved in H2O2 production (e.g., pyruvate oxidase, NADH oxidase, and lactate oxidase) did not affect this. Here we describe the purification of a novel NADH-dependent flavin reductase encoded by two highly similar genes (LJ_0548 and LJ_0549) that are conserved in lactobacilli belonging to the Lactobacillus acidophilus group. The genes are predicted to encode two 20-kDa proteins containing flavin mononucleotide (FMN) reductase conserved domains. Reductase activity requires FMN, flavin adenine dinucleotide (FAD), or riboflavin and is specific for NADH and not NADPH. The Km for FMN is 30 ± 8 μM, in accordance with its proposed in vivo role in H2O2 production. Deletion of the encoding genes in L. johnsonii led to a 40-fold reduction of hydrogen peroxide formation. H2O2 production in this mutant could only be restored by in trans complementation of both genes. Our work identifies a novel, conserved NADH-dependent flavin reductase that is prominently involved in H2O2 production in L. johnsonii. PMID:24487531

  13. Carrageenan :the difference between PNG and KCL gel precipitation method as Lactobacillus acidophilus encapsulation material

    NASA Astrophysics Data System (ADS)

    Setijawati, D.; Nursyam, H.; Salis, H.

    2018-04-01

    The study on the effects of using of materials and methods in the preparation of the microcapsules Lactobacillus acidophilus towards the viability has been done. The research method used is experimental laboratory design. Variable research was kind of material (A) as the first factor with sub factor (A1 = Eucheuma cottonii) (A2 = Eucheuma spinosum) (A3 = mixture of Eucheuma cottonii and Eucheuma spinosum 1:1 ratio), while the second factor is a method of extraction to produce caragenan (B) with sub factor (B1 = Philipine Natural Grade modification) (B2 = KCl gel Press Precipitation). Analysis of different influences uses Analysis Of Varians followed by Fisher’s test. Analysis of data uses Mini tab 16. The results shows that the kind of extraction factors and methods gave significantly different effects on the viability of Lactobacillus acidophilus. The highest mean of Viablity obtained in the treatment of materials with a mixture of Eucheuma cottonii and Eucheuma spinosum and used KCl Gel Press method is equal to 7.14 log (CFU / mL). It is ssuggested using of kappa-iota carrageenanmixture asencapsulation material with KCl Gel Press method on Lactobacillus acidophilus microencapsulation process because it treatment gavethe highest average of Lactobacillus acidophilus viability.

  14. Optimization of the medium for Lactobacillus acidophilus by Plackett-Burman and steepest ascent experiment.

    PubMed

    Chen, He; Niu, Jinfeng; Qin, Tao; Ma, Qi; Wang, Lei; Shu, Guowei

    2015-01-01

    Lactobacillus acidophilus not only improves the intestinal flora balance but also inhabits the growth of undesirable microorganisms in intestine, which is benefit to the health of humans and animals. Plackett-Burman and steepest ascent experiment are the rapid and concise ways of screening the main effective factors. This study is aimed to select the main influence factors and optimize the medium for Lactobacillus acidophilus by Plackett-Burman experiment and steepest ascent experiment. The ideal carbon source was screened among glucose, maltose, lactose and whey powder, and the ideal nitrogen source was screened among casein hydrolysate, peptone, yeast extract powder, fish meal, carbamide, ammonium sulfate and sodium nitrate by single factor experiment. Plackett-Burman and steepest ascent experiment were applied to screen the main effective factors of Lactobacillus acidophilus among peptone, beef extract, yeast extract powder, glucose, K2HPO4, C6H14O7N2, CH3COONa, MgSO4 and Tween-80. Result. The results indicated that glucose (p = 0.01510) as negative factor and K2HPO4 (p = 0.02017) as positive effect were the significant growth factors of Lactobacillus acidophilus, CH3COONa (p = 0.09273) as positive effect was an important factor, and the optimized medium was as follows: glucose - 21 g/L, K2HPO4 - 3.5 g/L, CH3COONa - 6.5 g/L, peptone - 10 g/L, beef extract - 8 g/L, yeast extract pow. nd. Lactobacillus acidophilus not only improves the intestinal flora balance but also inhabits the growth of undesirable microorganisms in intestine, which is benefit to the health of humans and animals. Plackett-Burman and steepest ascent experiment are the rapid and concise ways of screening the main effective factors. This study is aimed to select the main influence factors and optimize the medium for Lactobacillus acidophilus by Plackett-Burman experiment and steepest ascent experiment. Material and methods. The ideal carbon source was screened among glucose, maltose, lactose and

  15. 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.

  16. Proteomics Analysis of the Adhesion Activity of Lactobacillus acidophilus ATCC 4356 Upon Growth in an Intestine-Like pH Environment.

    PubMed

    Wu, Zhen; Wang, Gang; Wang, Wenwen; Pan, Daodong; Peng, Liuyang; Lian, Liwei

    2018-03-01

    Many health effects of Lactobacillus acidophilus are desirable among these the adhesion ability is vital to enhance the possibility of colonization and stabilization associated with the gut mucosal barrier. In this study, the growth characteristics and the adhesion activity of L. acidophilus in the intestine-like pH environment (pH 7.5) are identified. The number of bacteria adhering to the HT-29 cells is found with a gradual increase trend (pH 5.5-7.5). This also leads to the morphological changes of L. acidophilus after exposure to different pH environments. Furthermore, with the help of the isobaric tags for relative and absolute quantification (iTRAQ) proteomic analysis, 207 proteins are detected differentially expressed at pH of 7.5. The use of GO analysis and KEGG analysis indicates three essential pathways related to the cell envelope peptide-glycan biosynthesis, carbohydrate metabolism, and amino acid metabolism are obviously changed. Adhesion related surface protein fmtB and PrtP are upregulated in pH 7.5 group. While the moonlight proteins like pyruvate kinase, which binds specifically to the mucin layer and inhibits the adhesive activity of L. acidophilus, is found downregulated. These results could be useful to understand the adhesion mechanism of L. acidophilus adapting for the gut mucosal barrier in the intestinal environment. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Lactobacillus acidophilus Induces Cytokine and Chemokine Production via NF-κB and p38 Mitogen-Activated Protein Kinase Signaling Pathways in Intestinal Epithelial Cells

    PubMed Central

    Lü, Xuena; Man, Chaoxin; Han, Linlin; Shan, Yi; Qu, Xingguang; Liu, Ying; Yang, Shiqin; Xue, Yuqing; Zhang, Yinghua

    2012-01-01

    Intestinal epithelial cells can respond to certain bacteria by producing an array of cytokines and chemokines which are associated with host immune responses. Lactobacillus acidophilus NCFM is a characterized probiotic, originally isolated from human feces. This study aimed to test the ability of L. acidophilus NCFM to stimulate cytokine and chemokine production in intestinal epithelial cells and to elucidate the mechanisms involved in their upregulation. In experiments using intestinal epithelial cell lines and mouse models, we observed that L. acidophilus NCFM could rapidly but transiently upregulate a number of effector genes encoding cytokines and chemokines such as interleukin 1α (IL-1α), IL-1β, CCL2, and CCL20 and that cytokines showed lower expression levels with L. acidophilus NCFM treatment than chemokines. Moreover, L. acidophilus NCFM could activate a pathogen-associated molecular pattern receptor, Toll-like receptor 2 (TLR2), in intestinal epithelial cell lines. The phosphorylation of NF-κB p65 and p38 mitogen-activated protein kinase (MAPK) in intestinal epithelial cell lines was also enhanced by L. acidophilus NCFM. Furthermore, inhibitors of NF-κB (pyrrolidine dithiocarbamate [PDTC]) and p38 MAPK (SB203580) significantly reduced cytokine and chemokine production in the intestinal epithelial cell lines stimulated by L. acidophilus NCFM, suggesting that both NF-κB and p38 MAPK signaling pathways were important for the production of cytokines and chemokines induced by L. acidophilus NCFM. PMID:22357649

  18. Lactobacillus acidophilus contributes to a healthy environment for vaginal epithelial cells.

    PubMed

    Pi, Woojin; Ryu, Jae-Sook; Roh, Jaesook

    2011-09-01

    Lactobacillus species in the female genital tract are thought to act as a barrier to infection. Several studies have demonstrated that lactobacilli can adhere to vaginal epithelial cells. However, little is known about how the adherence of lactobacilli to vaginal epithelial cells affects the acidity, cell viability, or proliferation of the lactobacilli themselves or those of vaginal epithelial cells. Lactobacillus acidophilus was co-cultured with immortalized human vaginal epithelial cells (MS74 cell line), and the growth of L. acidophilus and the acidity of the culture medium were measured. MS74 cell density and viability were also assessed by counting cell numbers and observing the cell attachment state. L. acidophilus showed exponential growth for the first 6 hr until 9 hr, and the pH was maintained close to 4.0-5.0 at 24 hr after culture, consistent with previous studies. The growth curve of L. acidophilus or the pH values were relatively unaffected by co-culture with MS74 cells, confirming that L. acidophilus maintains a low pH in the presence of MS74 cells. This co-culture model could therefore potentially be used to mimic vaginal conditions for future in vitro studies. On the other hand, MS74 cells co-cultured with L. acidophilus more firmly attached to the culture plate, and a higher number of cells were present compared to cells cultured in the absence of L. acidophilus. These results indicate that L. acidophilus increases MS74 cell proliferation and viability, suggesting that lactobacilli may contribute to the healthy environment for vaginal epithelial cells.

  19. Effect of Lactobacillus acidophilus and Bifidobacterium bifidum supplementation to standard triple therapy on Helicobacter pylori eradication and dynamic changes in intestinal flora.

    PubMed

    Wang, Yu-huan; Huang, Ying

    2014-03-01

    To investigate Lactobacillus acidophilus (L. acidophilus) and Bifidobacterium bifidum (B. bifidum) supplementation to triple therapy for Helicobacter pylori (H. pylori) eradication and dynamic changes in intestinal flora in children with H. pylori infection. One hundred H. pylori-infected children were randomly assigned to two groups: treatment group (n = 43), standard triple anti-H. pylori therapy plus probiotics of L. acidophilus and B. bifidum for 2 weeks followed by taking probiotics for another 4 weeks; control group (n = 45), standard triple anti-H. pylori therapy for 6 weeks. After 6-week treatment, ¹³C-urease breath test was performed and side effects were monitored during the observation period. Quantitative PCR with 16S rRNA-gene-targeted species-specific primers was carried out for the analysis of human intestinal B. bifidum, L. acidophilus, and Escherichia coli (E. coli). As expected, treatment group could significantly enhance the H. pylori eradication rate (83.7 vs. 64.4 %, P < 0.05). B. bifidum, L. acidophilus, and E. coli showed no statistical difference before or after therapy in the treatment group. The number of B. bifidum and L. acidophilus was significantly decreased after 2-week treatment in the control group, but after 6-week treatment it significantly increased and nearly returned to the level before treatment. The number of E. coli increased significantly after 2-week treatment, while after 6-week treatment, it nearly decreased to the level before treatment. L. acidophilus and B. bifidum supplementation is effective for H. pylori eradication compared with triple therapy alone.

  20. AcmB Is an S-Layer-Associated β-N-Acetylglucosaminidase and Functional Autolysin in Lactobacillus acidophilus NCFM

    PubMed Central

    Johnson, Brant R.

    2016-01-01

    ABSTRACT Autolysins, also known as peptidoglycan hydrolases, are enzymes that hydrolyze specific bonds within bacterial cell wall peptidoglycan during cell division and daughter cell separation. Within the genome of Lactobacillus acidophilus NCFM, there are 11 genes encoding proteins with peptidoglycan hydrolase catalytic domains, 9 of which are predicted to be functional. Notably, 5 of the 9 putative autolysins in L. acidophilus NCFM are S-layer-associated proteins (SLAPs) noncovalently colocalized along with the surface (S)-layer at the cell surface. One of these SLAPs, AcmB, a β-N-acetylglucosaminidase encoded by the gene lba0176 (acmB), was selected for functional analysis. In silico analysis revealed that acmB orthologs are found exclusively in S-layer- forming species of Lactobacillus. Chromosomal deletion of acmB resulted in aberrant cell division, autolysis, and autoaggregation. Complementation of acmB in the ΔacmB mutant restored the wild-type phenotype, confirming the role of this SLAP in cell division. The absence of AcmB within the exoproteome had a pleiotropic effect on the extracellular proteins covalently and noncovalently bound to the peptidoglycan, which likely led to the observed decrease in the binding capacity of the ΔacmB strain for mucin and extracellular matrices fibronectin, laminin, and collagen in vitro. These data suggest a functional association between the S-layer and the multiple autolysins noncovalently colocalized at the cell surface of L. acidophilus NCFM and other S-layer-producing Lactobacillus species. IMPORTANCE Lactobacillus acidophilus is one of the most widely used probiotic microbes incorporated in many dairy foods and dietary supplements. This organism produces a surface (S)-layer, which is a self-assembling crystalline array found as the outermost layer of the cell wall. The S-layer, along with colocalized associated proteins, is an important mediator of probiotic activity through intestinal adhesion and modulation of

  1. AcmB Is an S-Layer-Associated β-N-Acetylglucosaminidase and Functional Autolysin in Lactobacillus acidophilus NCFM.

    PubMed

    Johnson, Brant R; Klaenhammer, Todd R

    2016-09-15

    Autolysins, also known as peptidoglycan hydrolases, are enzymes that hydrolyze specific bonds within bacterial cell wall peptidoglycan during cell division and daughter cell separation. Within the genome of Lactobacillus acidophilus NCFM, there are 11 genes encoding proteins with peptidoglycan hydrolase catalytic domains, 9 of which are predicted to be functional. Notably, 5 of the 9 putative autolysins in L. acidophilus NCFM are S-layer-associated proteins (SLAPs) noncovalently colocalized along with the surface (S)-layer at the cell surface. One of these SLAPs, AcmB, a β-N-acetylglucosaminidase encoded by the gene lba0176 (acmB), was selected for functional analysis. In silico analysis revealed that acmB orthologs are found exclusively in S-layer- forming species of Lactobacillus Chromosomal deletion of acmB resulted in aberrant cell division, autolysis, and autoaggregation. Complementation of acmB in the ΔacmB mutant restored the wild-type phenotype, confirming the role of this SLAP in cell division. The absence of AcmB within the exoproteome had a pleiotropic effect on the extracellular proteins covalently and noncovalently bound to the peptidoglycan, which likely led to the observed decrease in the binding capacity of the ΔacmB strain for mucin and extracellular matrices fibronectin, laminin, and collagen in vitro These data suggest a functional association between the S-layer and the multiple autolysins noncovalently colocalized at the cell surface of L. acidophilus NCFM and other S-layer-producing Lactobacillus species. Lactobacillus acidophilus is one of the most widely used probiotic microbes incorporated in many dairy foods and dietary supplements. This organism produces a surface (S)-layer, which is a self-assembling crystalline array found as the outermost layer of the cell wall. The S-layer, along with colocalized associated proteins, is an important mediator of probiotic activity through intestinal adhesion and modulation of the mucosal immune

  2. Selective and differential enumerations of Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus casei and Bifidobacterium spp. in yoghurt--a review.

    PubMed

    Ashraf, Rabia; Shah, Nagendra P

    2011-10-03

    Yoghurt is increasingly being used as a carrier of probiotic bacteria for their potential health benefits. To meet with a recommended level of ≥10(6) viable cells/g of a product, assessment of viability of probiotic bacteria in market preparations is crucial. This requires a working method for selective enumeration of these probiotic bacteria and lactic acid bacteria in yoghurt such as Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lb. acidophilus, Lb. casei and Bifidobacterium. This chapter presents an overview of media that could be used for differential and selective enumerations of yoghurt bacteria. De Man Rogosa Sharpe agar containing fructose (MRSF), MRS agar pH 5.2 (MRS 5.2), reinforced clostridial prussian blue agar at pH 5.0 (RCPB 5.0) or reinforced clostridial agar at pH 5.3 (RCA 5.3) are suitable for enumeration of Lb. delbrueckii subsp. bulgaricus when the incubation is carried out at 45°C for 72h. S. thermophilus (ST) agar and M17 are recommended for selective enumeration of S. thermophilus. Selective enumeration of Lb. acidophilus in mixed culture could be made in Rogosa agar added with 5-bromo-4-chloro-3-indolyl-β-d-glucopyranoside (X-Glu) or MRS containing maltose (MRSM) and incubation in a 20% CO2 atmosphere. Lb. casei could be selectively enumerated on specially formulated Lb. casei (LC) agar from products containing yoghurt starter bacteria (S. thermophilus and Lb. delbrueckii subsp. bulgaricus), Lb. acidophilus, Bifidobacterium spp. and Lb. casei. Bifidobacterium could be enumerated on MRS agar supplemented with nalidixic acid, paromomycin, neomycin sulphate and lithium chloride (MRS-NPNL) under anaerobic incubation at 37°C for 72h. Copyright © 2011. Published by Elsevier B.V.

  3. Lactobacillus acidophilus modulates inflammatory activity by regulating the TLR4 and NF-κB expression in porcine peripheral blood mononuclear cells after lipopolysaccharide challenge.

    PubMed

    Lee, Sang In; Kim, Hyun Soo; Koo, Jin Mo; Kim, In Ho

    2016-02-28

    A total of forty weaned pigs ((Landrace × Yorkshire) × Duroc) were used to evaluate the effects of Lactobacillus acidophilus on inflammatory activity after lipopolysaccharide (LPS) challenge. Experimental treatments were as follows: (T1) control diet+saline challenge; (T2) control diet with 0·1% L. acidophilus+saline challenge; (T3) control diet+LPS challenge; and (T4) control diet with 0·1% L. acidophilus+LPS challenge. On d-14, piglets were challenged with saline (T1 and T2) or LPS (T3 and T4). Blood samples were obtained at 0, 2, 4, 6 and 12 h after being challenged and analysed for immune cell cytokine production and gene expression pattern. The L. acidophilus treatment increased the average daily weight gain (ADWG) and average daily feed intake (ADFI) compared with the control diet. With the control diet, the LPS challenge (T3) increased the number of immune cells and expression of TNF-α and IL-6 compared with the saline challenge (T1). Whereas with the saline challenge L. acidophilus treatment (T2) increased the number of leucocytes and CD4 compared with the control diet (T1), with the LPS challenge L. acidophilus treatment (T4) decreased the number of leucocytes, lymphocytes, CD4+ and CD8+ and expression of TNF-α and IL-6 compared with the control diet (T3). L. acidophilus treatment decreased the expression of TRL4 and NF-κB in peripheral blood mononuclear cells (PBMC) after LPS challenge, which leads to inhibition of TNF-α, IFN-γ, IL-6, IL-8 and IL1B1 and to induction of IL-4 and IL-10. We suggested that L. acidophilus improved ADWG and ADFI and protected against LPS-induced inflammatory responses by regulating TLR4 and NF-κB expression in porcine PBMC.

  4. Comparison of the antimicrobial efficacy of photodynamic therapy with two mediators against Lactobacillus acidophilus in vitro.

    PubMed

    Azizi, Arash; Mousavian, Shiva; Taheri, Soudabeh; Lawaf, Shirin; Gonoudi, Elnaz; Rahimi, Arash

    2018-03-01

    Lactobacillus is a cariogenic microorganism. Different therapeutic approaches including photodynamic therapy (PDT) have been suggested for treatment of bacterial infection. The purpose of the current study was to compare the effects of PDT with Indocyanine green (ICG) and Methylene blue (MB) photosensitizers (PSs) on Lactobacillus acidophilus (L. acidophilus). In this in-vitro experimental study, 84 samples of L. acidophilus (1 McFarland standard) were compared in 14 experimental groups including: MB, ICG, 660-nm laser, 808-nm laser (pulsed, 74s/continuous-wave, 37s), different combinations of lasers and PSs, Chlorhexidine (CHX) 0.2%, sodium hypochlorite (NaOCl) 2.5%, penicillin 6.3.3 and control groups. The samples were cultured in microplates containing blood agar culture medium. After incubation at 37 °C for 48 h, the colony forming units (CFUs) of L. acidophilus were counted and compared before and after therapeutic interventions. Data were analyzed using SPSS19 software program according to one-way ANOVA test. This study showed that the separate use of ICG, 660- and 808-nm lasers (pulsed, 74s/continuous-wave, 37s), and the combined use of 808-nm laser (pulsed, 74s/continuous-wave, 37s) and ICG have no significant inhibitory effect on L. acidophilus colonies (P > 0.05), whereas the separate use of MB and the combined use of 660-nm laser (continuous-wave, 37s/pulsed, 74s) and MB significantly inhibited the growth of L. acidophilus in comparison with the control group (p < 0.05). Likewise, CHX 0.2%, NaOCl 2.5% and penicillin 6.3.3 significantly inhibited the bacterial growth (p < 0.05). The results showed that separate use of MB and combined use of 660-nm laser and MB have a significant inhibitory effect on L. acidophilus growth. Copyright © 2018 Elsevier B.V. All rights reserved.

  5. Survival of Lactobacillus acidophilus and Bifidobacterium bifidum in ice cream for use as a probiotic food.

    PubMed

    Hekmat, S; McMahon, D J

    1992-06-01

    Probiotic ice cream was made by fermenting a standard ice cream mix with Lactobacillus acidophilus and Bifidobacterium bifidum cultures and then freezing the mix in a batch freezer. Survival of the L. acidophilus and B. bifidum, as well as beta-galactosidase activity, was monitored during 17 wk of frozen storage at -29 degrees C. After freezing of the fermented mix, bacterial counts were 1.5 x 10(8) cfu/ml for L. acidophilus and 2.5 x 10(8) cfu/ml for B. bifidum. Seventeen weeks after freezing, these counts had decreased to 4 x 10(6) and 1 x 10(7) cfu/ml, respectively. During the same period, beta-galactosidase activity decreased from 1800 to 1300 units/ml. Probiotic ice cream was prepared at pH 5.0, 5.5, and 6.0 to determine consumer preferences and was compared with standard Utah State University "Aggie" ice cream. All samples were strawberry-flavored and were evaluated by 88 judges. The preferred pH of probiotic ice cream, based on overall acceptance, was pH 5.5. We demonstrated that probiotic ice cream is a suitable vehicle for delivering beneficial microorganisms such as L. acidophilus and B. bifidum to consumers. The bacteria can be grown to high numbers in ice cream mix and remain viable during frozen storage.

  6. Influence of the Probiotic Lactobacillus acidophilus NCFM
and Lactobacillus rhamnosus HN001 on Proteolysis Patterns
of Edam Cheese

    PubMed Central

    Cichosz, Grażyna; Nalepa, Beata; Kowalska, Marika

    2014-01-01

    Summary The objective of this study is to determine the viability of Lactobacillus acidophilus NCFM and Lactobacillus rhamnosus HN001 in Edam cheese as well as the effect of probiotic bacteria on paracasein proteolysis and changes in the water activity during ripening. The use of probiotics L. rhamnosus HN001 and L. acidophilus NCFM in Edam cheese slightly changed its chemical composition, but the change was not significant. The pH values were significantly correlated with the changes in Lactobacillus count (R=–0.807) and the level of phosphotungstic acid-soluble nitrogen compounds in total nitrogen (PTA-SN/TN) (R=0.775). After 10 weeks of ripening, the highest level of trichloroacetic acid-soluble nitrogen compounds in total nitrogen (TCA-SN/TN) was observed in the cheese containing L. rhamnosus HN001 (11.87%) and slightly lower level in the cheese containing L. acidophilus NCFM (7.60%) and control cheese (6.24%). The highest level of PTA-SN/TN fraction was noted in cheese containing L. acidophilus NCFM (3.48%) but the lowest level was observed in control cheese (2.24%) after ten weeks of ripening. The changes in the levels of PTA-SN/TN (R=–0.813) and TCA-SN/TN (R=–0.717) fractions were significantly (p<0.05) correlated with the viability of probiotic counts. Water activity (aw) strongly correlated with the PTA-SN/TN level (R=–0.824) and bacteria viability (R=–0.728). All of the analyzed cheeses were characterized by high counts of L. rhamnosus HN001 and L. acidophilus NCFM during ten weeks of ripening. PMID:27904317

  7. Effects of mannan-oligosaccharides and Lactobacillus acidophilus supplementation on growth performance, nutrient utilization and faecal characteristics in Murrah buffalo calves.

    PubMed

    Sharma, A N; Kumar, S; Tyagi, A K

    2018-06-01

    A study of 120 days was undertaken to ascertain the effect of mannan-oligosaccharides (MOS) and Lactobacillus acidophilus supplementation on growth performance, nutrient utilization and faecal characteristics in Murrah buffalo calves. Twenty Murrah buffalo calves of 5-7 days old and 31 ± 2.0 kg of body weight (BW) were randomly assigned into four groups. Group I served as the control (CON) in which only basal diet (concentrate mixture and green fodder) was provided, without any supplementation. Mannan-oligosaccharides at 4 g/calf/day were supplemented as prebiotic to Group II (PRE), whereas Group III (PRO) received Lactobacillus acidophilus in the form of fermented milk as probiotic at 200 ml/calf/day having 10 8  CFU/ml and Group IV (SYN) was supplemented with both MOS and Lactobacillus acidophilus as synbiotic at similar dose. Final BW (kg), dry matter intake, average daily gain, feed conversion efficiency and structural growth measurements were improved (p < .05) in the treatment groups compared to control. Digestibility of neutral detergent fibre was higher (p < .05) in SYN followed by PRE and PRO than control. The faecal lactobacilli and bifidobacterium population was higher (p < .05) in all the supplemented groups with a concomitant reduction in faecal coliform count as compared to control. Faecal ammonia, lactate and pH were also altered favourably (p < .05) in all the supplemented groups as compared to CON. The faecal volatile fatty acids were higher (p < .05) in PRE, PRO and SYN group than CON. The incorporation of MOS and Lactobacillus acidophilus in diet either individually or in combination as synbiotic has the potential to improve the performance and faecal characteristics in Murrah buffalo calves; however, the observed responses among the treatment groups were more evident in the synbiotic fed group compared to individual supplementation of MOS and Lactobacillus acidophilus. © 2018 Blackwell Verlag GmbH.

  8. 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.

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

    PubMed Central

    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

  10. Lactobacillus casei and Lactobacillus acidophilus regulate inflammatory pathway and improve antioxidant status in collagen-induced arthritic rats.

    PubMed

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

    2013-01-01

    In view of well-established immunomodulatory properties of Lactobacillus, present investigation was carried out to evaluate antioxidant and anti-inflammatory potential of Lactobacillus casei and Lactobacillus acidophilus, against inflammatory pathway and oxidative stress developed in an experimental model of arthritis. Collagen-induced arthritis (CIA) model was used. Oral administration of L. casei, L. acidophilus, standard antiarthritic drug indomethacin, and vehicle were started after induced arthritis and continued up to day 28. Interleukin (IL)-6, tumor necrosis factor (TNF)-α, IL-1β, IL-17, IL-4, and IL-10 levels were estimated in serum. In parallel, oxidative stress parameters were also measured from synovial effsuate. All rats were graded for arthritis score at the end of each week. L. casei, L. acidophilus, and indomethacin treatment significantly downregulated proinflammatory and upregulated anti-inflammatory cytokines at P<0.0001. They have significantly decreased oxidative stress in synovial effsuate (P<0.0001) and also arthritis score (P<0.05). Protection provided by L. casei and L. acidophilus was more pronounced than that of indomethacin. These lines of evidence suggest that L. casei and L. acidophilus exert potent protective effect against CIA. It further establishes effective anti-inflammatory and antioxidant properties of Lactobacillus. However, additional clinical investigations are needed to prove the efficacy of Lactobacillus in treatment/management of rheumatoid arthritis.

  11. 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.

  12. The domestication of the probiotic bacterium Lactobacillus acidophilus

    PubMed Central

    Bull, Matthew J.; Jolley, Keith A.; Bray, James E.; Aerts, Maarten; Vandamme, Peter; Maiden, Martin C. J.; Marchesi, Julian R.; Mahenthiralingam, Eshwar

    2014-01-01

    Lactobacillus acidophilus is a Gram-positive lactic acid bacterium that has had widespread historical use in the dairy industry and more recently as a probiotic. Although L. acidophilus has been designated as safe for human consumption, increasing commercial regulation and clinical demands for probiotic validation has resulted in a need to understand its genetic diversity. By drawing on large, well-characterised collections of lactic acid bacteria, we examined L. acidophilus isolates spanning 92 years and including multiple strains in current commercial use. Analysis of the whole genome sequence data set (34 isolate genomes) demonstrated L. acidophilus was a low diversity, monophyletic species with commercial isolates essentially identical at the sequence level. Our results indicate that commercial use has domesticated L. acidophilus with genetically stable, invariant strains being consumed globally by the human population. PMID:25425319

  13. The domestication of the probiotic bacterium Lactobacillus acidophilus.

    PubMed

    Bull, Matthew J; Jolley, Keith A; Bray, James E; Aerts, Maarten; Vandamme, Peter; Maiden, Martin C J; Marchesi, Julian R; Mahenthiralingam, Eshwar

    2014-11-26

    Lactobacillus acidophilus is a Gram-positive lactic acid bacterium that has had widespread historical use in the dairy industry and more recently as a probiotic. Although L. acidophilus has been designated as safe for human consumption, increasing commercial regulation and clinical demands for probiotic validation has resulted in a need to understand its genetic diversity. By drawing on large, well-characterised collections of lactic acid bacteria, we examined L. acidophilus isolates spanning 92 years and including multiple strains in current commercial use. Analysis of the whole genome sequence data set (34 isolate genomes) demonstrated L. acidophilus was a low diversity, monophyletic species with commercial isolates essentially identical at the sequence level. Our results indicate that commercial use has domesticated L. acidophilus with genetically stable, invariant strains being consumed globally by the human population.

  14. Inhibition of Insulin Degrading Enzyme and Insulin Degradation by UV-Killed Lactobacillus acidophilus.

    PubMed

    Neyazi, Nadia; Motevaseli, Elahe; Khorramizadeh, Mohammad Reza; Mohammadi Farsani, Taiebeh; Nouri, Zahra; Nasli Esfahani, Ensieh; Ghahremani, Mohammad Hossein

    2018-05-11

    Probiotics have beneficial effects on management of type 2 diabetes (T2D). The major hallmarks of T2D are insulin deficiency and insulin resistance which emphasize insulin therapy in onset of disease. Lactobacilli such as Lactobacillus acidophilus ( L. acidophilus ) have well known properties on prevention of T2D and insulin resistance but not on insulin degradation. Insulin-degrading enzyme (IDE) degrades insulin in the human body. We studied the effects of cell-free supernatant (CFS) and ultraviolet (UV)-killed L. acidophilus (ATCC 314) on IDE activity and insulin degradation in vitro. Cell growth inhibition by CFS and UV-killed L. acidophilus (ATCC 314) was studied and Western blotting and a fluoregenic assay was performed to determine IDE expression and its activity, respectively. Insulin degradation was evaluated by sandwich enzyme-linked immunosorbent assay(ELISA). IDE expression and activity was reduced by CFS and UV-killed L. acidophilus (ATCC 314). Although, decreased enzyme expression and activity was not significant for CFS in contrast to MRL (MRS with same pH as CFS). Also, reduction in IDE activity was not statistically considerable when compared to IDE expression. Insulin degradation was increased by CFS but decreased by UV-killed L. acidophilus (ATCC 314).

  15. Short communication: Nutrient consumption patterns of Lactobacillus acidophilus KLDS 1.0738 in controlled pH batch fermentations.

    PubMed

    Lv, Xuepeng; Liu, Gefei; Sun, Xiaomei; Chen, Hongyu; Sun, Jiahui; Feng, Zhen

    2017-07-01

    This work focused on elucidating the nutrient consumption patterns of Lactobacillus acidophilus to guide the design of media for high-cell-density culture. We investigated the nutrient consumption patterns of L. acidophilus KLDS 1.0738 in chemically defined media in controlled pH batch fermentations. The most abundantly consumed amino acids, vitamins, ions, and purines and pyrimidines were Glu and Gly, pyridoxine and nicotinamide, K + and PO 4 3- , and guanine and uracil, respectively. The highest consumption rates for amino acids, vitamins, ions, and purines and pyrimidines were Asp and Arg, folic acid and pyridoxine, Fe 2+ and Mn 2+ , and uracil and thymine, respectively. Furthermore, most of the amino acids, as well as guanine, thymine, pyridoxine, folic acid, nicotinamide, Mg 2+ , PO 4 3- , and K + had the highest bioavailability from the end of the lag growth phase to the mid-exponential growth phase. The overall consumption of glucose, adenine nucleotides, 2'-deoxyguanosine monohydrate, calcium pantothenate, Fe 2+ and Mn 2+ decreased with increasing average growth rate, indicating more effective use of these nutritional components at a higher average growth rate, as biomass yield based on nutritional component consumption increased. Our findings help to formulate complex media for high-cell-density cultivation and provide a theoretical basis for L. acidophilus feeding strategies. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  16. Antibacterial effect of propolis derived from tribal region on Streptococcus mutans and Lactobacillus acidophilus: An in vitro study.

    PubMed

    Airen, Bhuvnesh; Sarkar, Priyanka Airen; Tomar, Urvashi; Bishen, Kundendu Arya

    2018-01-01

    The study aimed at investigating in vitro antimicrobial activity of ethanolic extract of propolis (EEP) and water extract of propolis against two main cariogenic oral pathogens: Streptococcus mutans and Lactobacillus acidophilus. Propolis was obtained from beehives in the Jhabua region of India. Ethanolic and water extracts were prepared at concentrations of 5% and 20% weight/volume (w/v). To support the results, a positive control (chlorhexidine 0.2%) and a negative control (distilled water) were used. S. mutans was cultured on brain-heart infusion agar and L. acidophilus was cultured on De Man, Rogosa, and Sharpe agar. The results showed that at concentrations of 5% and 20%, EEP was effective against S. mutans and L. acidophilus. However, at similar concentrations, water extract was effective only against L. acidophilus. The highest activity was shown by chlorhexidine (0.2%) with mean zones of inhibition of 13.9 mm and 15.1 mm against S. mutans and L. acidophilus, respectively. It can be concluded that the propolis extracted from tribal regions of Jhabua possesses antibacterial efficacy against S. mutans and L. acidophilus.

  17. Effect of complexation conditions on microcapsulation of Lactobacillus acidophilus in xanthan-chitosan polyelectrolyte complex gels.

    PubMed

    Chen, He; Song, Yajuan; Liu, Nina; Wan, Hongchang; Shu, Guowei; Liao, Na

    2015-01-01

    Lactobacillus acidophilus has become increasingly popular because of their beneficial effects on health of their host, and are called proboscis. In order to exert beneficial effects for probiotics, they must be able to tolerate the acidic conditions of the stomach environment and the bile in the small intestine. Microencapsulated form has received reasonable attention, since it can protect probiotic organisms against an unfavourable environment, and to allow their release in a viable and metabolically active state in the intestine. The aim of this study was to investigate some factores, such as chitosan solution pH and concentration, xanthan concentration, cell suspension-xanthan ratio, mixed bacteria glue liquid-chitosan ratio, which impacted the process of microencapsulation of L. acidophilus. In this study, L. acidophilus was immobilized with xanthan⁄chitosan gel using extrusion method. The viable counts and encapsulation yield of L. acidophilus encapsulated in different chitosan solution pH (4.5, 5, 5.5 and 6), in different chitosan concentration (0.5%, 0.7%, 0.9% and 1.1%), in different xanthan concentration (0.5%, 0.7%, 0.9% and 1.1%), in different cell suspension-xanthan ratios (1:5, 1:10, 1:15 and 1:20), in different mixed bacteria glue liquid-chitosan ratios (1:3, 1:4, 1:5 and 1:6), have been investigated by single factor experiment method. The optimum conditions of microencapsulated L. acidophilus have been observed. The optimum chitosan solution pH for L. acidophilus was 5.5; the optimum chitosan concentration was 0.9%; the optimum xanthan concentration was 0.7%; the optimum cell suspension-xanthan ratio was 1:10; the optimum mixed bacteria glue liquid-chitosan ratio was 1:3. These results will be helpful to further optimize the process of L. acidophilus microencapsulation, and provide reference for obtaining higher viable counts and entrapped yield of L. acidophilus microcapsules.

  18. The life history of Lactobacillus acidophilus as a probiotic: a tale of revisionary taxonomy, misidentification and commercial success.

    PubMed

    Bull, Matthew; Plummer, Sue; Marchesi, Julian; Mahenthiralingam, Eshwar

    2013-12-01

    Lactobacillus acidophilus is a commercially significant bacterial probiotic, originally isolated from the human gastrointestinal tract and designated Bacillus acidophilus in 1900. Throughout the development of methods to identify and characterise bacteria, L. acidophilus has undergone multiple taxonomic revisions and is now the type species of a phylogenetic subgroup in the highly diverse and heterogeneous Lactobacillus genus. As a result of the limitations of differentiating phenotypically similar species by morphological and biochemical means and revisionary nature of Lactobacillus taxonomy, the characterisation of L. acidophilus has struggled with misidentification and misrepresentation. In contrast, due to its global use as a probiotic supplement in functional foods, L. acidophilus sensu stricto is now one of the most well-characterised Lactobacillus species. Here, we establish the provenance of L. acidophilus strains, unpicking historical and current misidentifications of L. acidophilus, and reviewing the probiotic, genomic and physiological characteristics of this important Lactobacillus species. © 2013 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  19. Effect of Eudragit S100 nanoparticles and alginate chitosan encapsulation on the viability of Lactobacillus acidophilus and Lactobacillus rhamnosus.

    PubMed

    Ansari, Fereshteh; Pourjafar, Hadi; Jodat, Vahid; Sahebi, Javad; Ataei, Amir

    2017-12-01

    In this study, we examined a novel method of microencapsulation with calcium alginate-chitosan and Eudragit S100 nanoparticles for the improving viability of probiotic bacteria, Lactobacillus acidophilus and Lactobacillus rhamnosus. Extrusion technique was carried out in microencapsulation process. The viability of two probiotics in single coated beads (with only chitosan), double coated beads (with chitosan and Eudragit nanoparticles), and as free cells (unencapsulated) were conducted in simulated gastric juice (pH 1.55, without pepsin) followed by incubation in simulated intestinal juice (pH 7.5, with 1% bile salt). In case of single coated beads, presumably, lack of sufficient strength of chitosan under simulated gastric condition was the main reason of 4-log and 5-log reduction of the counts of the L. acidophilus and L. rhamnosus respectively. The results showed that with the second coat forming (Eudragit nanoparticles) over the first coat (chitosan), the strength of the beads and then viability rate of the bacteria were increased in comparison with the single coated beads.

  20. Use of Lactobacillus acidophilus and Lactobacillus casei for a potential probiotic legume-based fermented product using pigeon pea (Cajanus cajan).

    PubMed

    Parra, K; Ferrer, M; Piñero, M; Barboza, Y; Medina, L M

    2013-02-01

    The aim of the present study was to evaluate the use of pigeon pea (Cajanus cajan) as an appropriate substrate in the production of a legume-based fermented product with Lactobacillus acidophilus ATCC 314 or Lactobacillus casei ATCC 393 and then to ascertain the effects of the addition of ingredients such as powdered milk and banana or strawberry sauce. The products were analyzed for viable cell counts, pH, and sensory attributes during product manufacture and throughout the refrigerated storage period at 3, 7, 14, 21, and 28 days. Nine types of products were produced. At the end of the storage period, the viability of L. acidophilus was above 7 log CFU/g in the presence of milk and 20% sucrose fruit sauce. For products with L. casei, the lack of ingredients such as milk caused no significant loss in viability; however, a high concentration of sucrose in the fruit sauce was an important factor in maintaining a high L. casei population. L. casei had high viability and good sensory attributes. Both strains could be considered suitable for a pigeon pea-based fermented potential probiotic product and a low-cost protein source.

  1. 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

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

    PubMed

    Sun, Bo; Hu, Chen; 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.

  3. Simulated microgravity affects some biological characteristics of Lactobacillus acidophilus.

    PubMed

    Shao, Dongyan; Yao, Linbo; Riaz, Muhammad Shahid; Zhu, Jing; Shi, Junling; Jin, Mingliang; Huang, Qingsheng; Yang, Hui

    2017-04-01

    The effects of weightlessness on enteric microorganisms have been extensively studied, but have mainly been focused on pathogens. As a major component of the microbiome of the human intestinal tract, probiotics are important to keep the host healthy. Accordingly, understanding their changes under weightlessness conditions has substantial value. This study was carried out to investigate the characteristics of Lactobacillus acidophilus, a typical probiotic for humans, under simulated microgravity (SMG) conditions. The results revealed that SMG had no significant impact on the morphology of L. acidophilus, but markedly shortened its lag phase, enhanced its growth rate, acid tolerance ability up to pH < 2.5, and the bile resistance at the bile concentration of <0.05%. SMG also decreased the sensitivity of L. acidophilus to cefalexin, sulfur gentamicin, and sodium penicillin. No obvious effect of SMG was observed on the adhesion ability of L. acidophilus to Caco-2 cells. Moreover, after SMG treatment, both the culture of L. acidophilus and its liquid phase exhibited higher antibacterial activity against S. typhimurium and S. aureus in a time-dependent manner. The SMG treatment also increased the in vitro cholesterol-lowering ability of L. acidophilus by regulating the expression of the key cholesterol metabolism genes CYP7A1, ABCB11, LDLR, and HMGCR in the HepG2 cell line. Thus, the SMG treatment did have considerable influence on some biological activities and characteristics of L. acidophilus related to human health. These findings provided valuable information for understanding the influence of probiotics on human health under simulated microgravity conditions, at least.

  4. Lactobacillus acidophilus binds to MUC3 component of cultured intestinal epithelial cells with highest affinity.

    PubMed

    Das, Jugal Kishore; Mahapatra, Rajani Kanta; Patro, Shubhransu; Goswami, Chandan; Suar, Mrutyunjay

    2016-04-01

    Lactobacillus strains have been shown to adhere to the mucosal components of intestinal epithelial cells. However, established in vitro adhesion assays have several drawbacks in assessing the adhesion of new Lactobacillus strains. The present study aimed to compare the adhesion of four different Lactobacillus strains and select the most adherent microbe, based on in silico approach supported by in vitro results. The mucus-binding proteins in Lactobacillus acidophilus, L. plantarum, L. brevis and L. fermentum were identified and their capacities to interact with intestinal mucin were compared by molecular docking analysis. Lactobacillus acidophilus had the maximal affinity of binding to mucin with predicted free energy of -6.066 kcal mol(-1) Further, in vitro experimental assay of adhesion was performed to validate the in silico results. The adhesion of L. acidophilus to mucous secreting colon epithelial HT-29 MTX cells was highest at 12%, and it formed biofilm with maximum depth (Z = 84 μm). Lactobacillus acidophilus was determined to be the most adherent strain in the study. All the Lactobacillus strains tested in this study, displayed maximum affinity of binding to MUC3 component of mucus as compared to other gastrointestinal mucins. These findings may have importance in the design of probiotics and health care management. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  5. A mild pulsed electric field condition that improves acid tolerance, growth, and protease activity of Lactobacillus acidophilus LA-K and Lactobacillus delbrueckii subspecies bulgaricus LB-12.

    PubMed

    Najim, N; Aryana, Kayanush J

    2013-06-01

    Pulsed electric field (PEF) processing involves the application of pulses of voltage for less than 1 s to fluid products placed between 2 electrodes. The effect of mild PEF on beneficial characteristics of probiotic bacteria Lactobacillus acidophilus and Lactobacillus delbrueckii ssp. bulgaricus is not clearly understood. The objective of this study was to determine the influence of mild PEF conditions on acid tolerance, growth, and protease activity of Lb. acidophilus LA-K and Lactobacillus delbrueckii ssp. bulgaricus LB-12. A pilot plant PEF system (OSU-4M; The Ohio State University, Columbus) was used. The PEF treatments were positive square unipolar pulse width of 3 µs, pulse period of 0.5s, electric field strength of 1 kV/cm, delay time of 20 µs, flow rate of 60 mL/min, and 40.5°C PEF treatment temperature. Both Lb. acidophilus LA-K and Lb. bulgaricus LB-12 subjected to mild PEF conditions were acid tolerant until the end of the 120 min of incubation, unlike the Lb. bulgaricus control, which was not acid tolerant after 30 min. The mild PEF-treated Lb. acidophilus LA-K and Lb. bulgaricus LB-12 reached the logarithmic phase of growth an hour earlier than the control. Mild PEF conditions studied significantly improved acid tolerance, exponential growth, and protease activity of both Lb. acidophilus LA-K and Lb. bulgaricus LB-12 compared with the control. The mild PEF conditions studied can be recommended for pretreating cultures to enhance these desirable attributes. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  6. 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

  7. Functional Analysis of an S-Layer-Associated Fibronectin-Binding Protein in Lactobacillus acidophilus NCFM

    PubMed Central

    Hymes, Jeffrey P.; Johnson, Brant R.; Barrangou, Rodolphe

    2016-01-01

    Bacterial surface layers (S-layers) are crystalline arrays of self-assembling proteinaceous subunits called S-layer proteins (Slps) that comprise the outermost layer of the cell envelope. Many additional proteins that are associated with or embedded within the S-layer have been identified in Lactobacillus acidophilus NCFM, an S-layer-forming bacterium that is widely used in fermented dairy products and probiotic supplements. One putative S-layer-associated protein (SLAP), LBA0191, was predicted to mediate adhesion to fibronectin based on the in silico detection of a fibronectin-binding domain. Fibronectin is a major component of the extracellular matrix (ECM) of intestinal epithelial cells. Adhesion to intestinal epithelial cells is considered an important trait for probiotic microorganisms during transit and potential association with the intestinal mucosa. To investigate the functional role of LBA0191 (designated FbpB) in L. acidophilus NCFM, an fbpB-deficient strain was constructed. The L. acidophilus mutant with a deletion of fbpB lost the ability to adhere to mucin and fibronectin in vitro. Homologues of fbpB were identified in five additional putative S-layer-forming species, but no homologues were detected in species outside the L. acidophilus homology group. PMID:26921419

  8. Eruca sativa might influence the growth, survival under simulated gastrointestinal conditions and some biological features of Lactobacillus acidophilus, Lactobacillus plantarum and Lactobacillus rhamnosus strains.

    PubMed

    Fratianni, Florinda; Pepe, Selenia; Cardinale, Federica; Granese, Tiziana; Cozzolino, Autilia; Coppola, Raffaele; Nazzaro, Filomena

    2014-10-01

    The growth and viability of three Lactobacillus strains, Lactobacillus acidophilus, Lactobacillus plantarum and Lactobacillus rhamnosus, after their passage through simulated gastric and pancreatic juices were studied as a function of their presence in the growth medium of rocket salad (Eruca sativa). The presence of E. sativa affected some of the biological properties of the strains. For example, L. acidophilus and L. plantarum worked more efficiently in the presence of E. sativa, increasing not only the antioxidant activity of the medium, but also their own antioxidant power and antimicrobial activity; L. rhamnosus was not affected in the same manner. Overall, the presence of vegetables might help to boost, in specific cases, some of the characteristics of lactobacilli, including antioxidant and antimicrobial power.

  9. Eruca sativa Might Influence the Growth, Survival under Simulated Gastrointestinal Conditions and Some Biological Features of Lactobacillus acidophilus, Lactobacillus plantarum and Lactobacillus rhamnosus Strains

    PubMed Central

    Fratianni, Florinda; Pepe, Selenia; Cardinale, Federica; Granese, Tiziana; Cozzolino, Autilia; Coppola, Raffaele; Nazzaro, Filomena

    2014-01-01

    The growth and viability of three Lactobacillus strains, Lactobacillus acidophilus, Lactobacillus plantarum and Lactobacillus rhamnosus, after their passage through simulated gastric and pancreatic juices were studied as a function of their presence in the growth medium of rocket salad (Eruca sativa). The presence of E. sativa affected some of the biological properties of the strains. For example, L. acidophilus and L. plantarum worked more efficiently in the presence of E. sativa, increasing not only the antioxidant activity of the medium, but also their own antioxidant power and antimicrobial activity; L. rhamnosus was not affected in the same manner. Overall, the presence of vegetables might help to boost, in specific cases, some of the characteristics of lactobacilli, including antioxidant and antimicrobial power. PMID:25275269

  10. Effects of yogurt starter cultures on the survival of Lactobacillus acidophilus.

    PubMed

    Ng, Elizabeth W; Yeung, Marie; Tong, Phillip S

    2011-01-31

    Recognized to confer health benefits to consumers, probiotics such as Lactobacillus acidophilus are commonly incorporated into fermented dairy products worldwide; among which yogurt is a popular delivery vehicle. To materialize most of the putative health benefits associated with probiotics, an adequate amount of viable cells must be delivered at the time of consumption. However, the loss in their viabilities during refrigerated storage has been demonstrated previously. This study focused on the effects of yogurt starter cultures on the survival of five strains of L. acidophilus, with emphases on low pH and acid production. Differential survival behavior between L. acidophilus strains was further analyzed. To this end, viable cell counts of L. acidophilus were determined weekly during 4°C storage in various types of yogurts made with Streptococcus thermophilus alone, L. delbrueckii ssp. bulgaricus alone, both species of the starter cultures, or glucono-delta-lactone (GDL). All yogurt types, except for pasteurized yogurts, were co-fermented with L. acidophilus. Yogurt filtrate was analyzed for the presence of any inhibitory substance and for the amount of hydrogen peroxide. Multiplication of L. acidophilus was not affected by the starter cultures as all strains reached high level on day 0 of the storage period. Throughout the 28-day storage period, cell counts of L. acidophilus PIM703 and SBT2062 remained steady (~6 × 10(7)CFU/g) in yogurts made with both starter cultures, whereas those of ATCC 700396 and NCFM were reduced by a maximum of 3 and 4.6 logs, respectively. When starter cultures were replaced by GDL, all strains survived well, suggesting that a low pH was not a critical factor dictating their survival. In addition, the filtrate collected from yogurts made with starter cultures appeared to have higher inhibitory activities against L. acidophilus than that made with GDL. The presence of viable starter cultures was necessary to adversely affect the

  11. Antimicrobial effects of Lactobacillus plantarum and Lactobacillus acidophilus against multidrug-resistant enteroaggregative Escherichia coli.

    PubMed

    Kumar, Manesh; Dhaka, Pankaj; Vijay, Deepthi; Vergis, Jess; Mohan, Vysakh; Kumar, Ashok; Kurkure, Nitin V; Barbuddhe, Sukhadeo B; Malik, S V S; Rawool, Deepak B

    2016-09-01

    The in vitro and in vivo antimicrobial effects of Lactobacillus plantarum and Lactobacillus acidophilus were evaluated individually and synergistically against multidrug-resistant enteroaggregative Escherichia coli (MDR-EAEC). In vitro evaluation of each probiotic strain when co-cultured with MDR-EAEC isolates revealed a reduction in MDR-EAEC counts (eosin-methylene blue agar) in a dose- and time-dependent manner: probiotics at a dose rate of 10(10) CFU inhibited MDR-EAEC isolates at 72 h post-inoculation (PI), whereas at lower concentrations (10(8) and 10(9) CFU) MDR-EAEC isolates were inhibited at 96 h PI. The synergistic antimicrobial effect of both probiotic strains (each at 10(10) CFU) was highly significant (P < 0.01) and inhibited the growth of MDR-EAEC isolates at 24 h PI. For in vivo evaluation, weaned mice were fed orally with 10(7) CFU of MDR-EAEC. At Day 3 post-infection, treated mice were fed orally with the probiotic strains (each at 10(10) CFU). Compared with the control, post-treatment a significant (P < 0.01) reduction in MDR-EAEC counts was observed in faeces by Day 2 and in intestinal tissues of treated mice by Days 3 and 4 as evidenced by plate count (mean 2.71 log and 2.27 log, respectively) and real-time PCR (mean 1.62 log and 1.57 log, respectively) methods. Histopathologically, comparatively mild changes were observed in the ileum and colon from Days 3 to 5 post-treatment with probiotics; however, from Day 6 the changes were regenerative or normal. These observations suggest that these probiotic strains can serve as alternative therapeutics against MDR-EAEC-associated infections in humans and animals. Copyright © 2016 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.

  12. Lactobacillus acidophilus Suppresses Colitis-Associated Activation of the IL-23/Th17 Axis

    PubMed Central

    Chen, Linlin; Zou, Yiyou; Peng, Jie; Lu, Fanggen; Yin, Yani; Li, Fujun; Yang, Junwen

    2015-01-01

    The aim of this paper is to determine the modulatory effects of Lactobacillus acidophilus on the IL-23/Th17 immune axis in experimental colitis. DSS-induced mouse models of UC were to be saline, hormones, and different concentrations of Lactobacillus acidophilus intervention. The expression of interleukin- (IL-) 17, tumor necrosis factor α (TNFα), IL-23, transforming growth factor β1 (TGFβ1), signal transducer and activator of transcription 3 (STAT3), and phosphorylated (p)-STAT3 was examined by RT-PCR, Western blotting, and immunohistochemical analysis. And the results showed that administration of L. acidophilus suppressed Th17 cell-mediated secretion of proinflammatory cytokine IL-17 through downregulation of IL-23 and TGFβ1 expression and downstream phosphorylation of p-STAT3. PMID:25973440

  13. Some low homogenization pressures improve certain probiotic characteristics of yogurt culture bacteria and Lactobacillus acidophilus LA-K.

    PubMed

    Muramalla, T; Aryana, K J

    2011-08-01

    Lactobacillus delbrueckii ssp. bulgaricus, Streptococcus salivarius ssp. thermophilus, and Lactobacillus acidophilus are dairy cultures widely used in the manufacture of cultured dairy products. Commonly used homogenization pressures in the dairy industry are 13.80 MPa or less. It is not known whether low homogenization pressures can stimulate bacteria to improve their probiotic characteristics. Objectives were to determine the effect of homogenization at 0, 3.45, 6.90, 10.34, and 13.80 MPa on acid tolerance, bile tolerance, protease activity, and growth of L. delbrueckii ssp. bulgaricus LB-12, S. salivarius ssp. thermophilus ST-M5, and L. acidophilus LA-K. The cultures were individually inoculated in cool autoclaved skim milk (4°C) and homogenized for 5 continuous passes. Growth and bile tolerance of samples were determined hourly for 10h of incubation. Acid tolerance was determined every 20 min for 120 min of incubation. Protease activity was determined at 0, 12, and 24h of incubation. All homogenization pressures studied improved acid tolerance of L. delbrueckii ssp. bulgaricus LB-12 but had no beneficial effect on protease activity and had negative effects on growth and bile tolerance. A pressure of 6.90 MPa improved acid tolerance, bile tolerance, and protease activity of S. salivarius ssp. thermophilus ST-M5, but none of the homogenization pressures studied had an effect on its growth. Homogenization pressures of 13.80 and 6.90 MPa improved acid tolerance and bile tolerance, respectively, of L. acidophilus LA-K but had no effect on protease activity and its growth. Some low homogenization pressures positively influenced some characteristics of yogurt culture bacteria and L. acidophilus LA-K. Culture pretreatment with some low homogenization pressures can be recommended for improvement of certain probiotic characteristics. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  14. Optimization of Lactobacillus acidophilus cultivation using taro waste and evaluation of its biological activity.

    PubMed

    Hsieh, Shu-Chen; Liu, Jui-Ming; Pua, Xiao-Hui; Ting, Yuwen; Hsu, Ren-Jun; Cheng, Kuan-Chen

    2016-03-01

    In this study, taro waste (TW) was utilized for Lactobacillus acidophilus BCRC 14079 cultivation and the anti-tumor and immune-modulatory properties of heat-killed cells (HKCs), cytoplasmic fraction (CF), and exopolysaccharide (EPS) were evaluated. The optimum liquefaction enzyme dosage, temperature, and time determined by Box-Behnken design response surface methodology (BBD-RSM) were 9 mL/L of α-amylase, 79.2 °C, and 5 h of reaction, respectively. The optimum temperature and reaction time for saccharification were determined as 60 °C and 3 h. The optimum medium, CGMY1 medium, constitutes of TW hydrolysate containing 37 g/L of glucose, 25 g/L of corn gluten meal (CGM), and 1 g/L of yeast extract (YE). Results of MTT assay showed that HKCs and EPS from CGM medium exhibited the highest anti-proliferative in HT-29 (IC50 of HKCs, 467.25 μg/mL; EPS, 716.10 μg/mL) and in Caco-2 cells (IC50 of EPS, 741.60 μg/mL). Luciferase-based NF-ΚB and COX-2 systems indicated HKCs from CGM medium stimulated the highest expression of luciferin in both systems. The luciferase activities by using 100 and 500 μg/mL of HKCs from CGM were 24.30- and 45.83-fold in NF-ΚB system and 11.54- and 4.93-fold in COX-2 system higher than the control. In conclusion, this study demonstrated the potential of TW medium for L. acidophilus cultivation and the production of non-viable probiotics with enhanced biological activities.

  15. Functional Analysis of an S-Layer-Associated Fibronectin-Binding Protein in Lactobacillus acidophilus NCFM.

    PubMed

    Hymes, Jeffrey P; Johnson, Brant R; Barrangou, Rodolphe; Klaenhammer, Todd R

    2016-05-01

    Bacterial surface layers (S-layers) are crystalline arrays of self-assembling proteinaceous subunits called S-layer proteins (Slps) that comprise the outermost layer of the cell envelope. Many additional proteins that are associated with or embedded within the S-layer have been identified in Lactobacillus acidophilus NCFM, an S-layer-forming bacterium that is widely used in fermented dairy products and probiotic supplements. One putative S-layer-associated protein (SLAP), LBA0191, was predicted to mediate adhesion to fibronectin based on the in silico detection of a fibronectin-binding domain. Fibronectin is a major component of the extracellular matrix (ECM) of intestinal epithelial cells. Adhesion to intestinal epithelial cells is considered an important trait for probiotic microorganisms during transit and potential association with the intestinal mucosa. To investigate the functional role of LBA0191 (designated FbpB) in L. acidophilus NCFM, an fbpB-deficient strain was constructed. The L. acidophilus mutant with a deletion off bpB lost the ability to adhere to mucin and fibronectin in vitro Homologues off bpB were identified in five additional putative S-layer-forming species, but no homologues were detected in species outside theL. acidophilus homology group. Copyright © 2016 Hymes et al.

  16. 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 α-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 α-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

  17. Lactobacillus acidophilus Mixture in Treatment of Children Hospitalized With Acute Diarrhea.

    PubMed

    Pinto, Jamie M; Petrova, Anna

    2016-11-01

    Despite unproven effectiveness, Lactobacillus acidophilus is a widely used probiotic in the treatment of pediatric diarrhea. In this report, we evaluated the association between length of stay (LOS) for 290 young children hospitalized with acute diarrhea and adjuvant therapy with a probiotic mixture containing 80% L acidophilus that was included in treatment for 22.4% of them. Overall, no association between LOS and use of L acidophilus was recorded after controlling for age, length of diarrhea symptoms, duration of intravenous fluids, and prior exposure to antibiotic. However, LOS was directly associated with use of L acidophilus in children with negative stool studies, and no such association was recorded in children with positive stool for rotavirus or other infections. We concluded that adjuvant therapy with L acidophilus mixture is not beneficial for young children hospitalized with acute diarrhea. © The Author(s) 2015.

  18. 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. PMID:26730335

  19. Effects of Lactobacillus acidophilus supplementation on growth performance, nutrient digestibility, fecal microbial and noxious gas emission in weaning pigs.

    PubMed

    Lan, Ruixia; Koo, Jinmo; Kim, Inho

    2017-03-01

    Antibiotics used as growth promoters in livestock have been banned in the European Union since 2006. Antibiotics alternatives have focused on probiotics, such as Lactobacillus acidophilus. The concentration of L. acidophilus is considered crucial for obtaining the desired effects. However, limited studies have been conducted to test the dose-dependent effects of L. acidophilus. Therefore, the present study aimed to test the dose-dependent effects of L. acidophilus on growth performance, nutrient digestibility, fecal microbial flora and fecal noxious gas emission in weaning pigs. Lactobacillus acidophilus supplementation increased (P < 0.05) average daily gain, average daily feed intake, apparent nutrient digestibility of dry matter, nitrogen and gross energy, and Lactobacillus counts compared to the basal diet treatment, and a linear effect (P < 0.05) was observed on those criteria. Escherichia coli counts and NH 3 emission were decreased (P < 0.05) by L. acidophilus supplementation, and a linear effect (P < 0.05) was observed on E. coli counts. These results suggest that L. acidophilus could be used as an antibiotic alternative by improving growth performance, nutrient digestibility and gut balance (i.e. increased Lactobacillus counts and decreased E. coli counts), and decreasing NH3 emission, of weaning pigs. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

  20. 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. Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  1. Characterization of adhesive molecule with affinity to Caco-2 cells in Lactobacillus acidophilus by proteome analysis.

    PubMed

    Ashida, Nobuhisa; Yanagihara, Sae; Shinoda, Tadashi; Yamamoto, Naoyuki

    2011-10-01

    The adhesive activities of eight Lactobacillus acidophilus strains toward intestinal epithelial Caco-2 cells were studied to understand the probiotic characteristics of the L. acidophilus L-92 strain. Most of the strains, including L-92, showed high adhesive activity; CP23 showed the lowest adhesive activity. CP23 was selected for comparative analysis of cell wall-associated proteins versus the L-92 strain. Cell wall-associated proteins extracted from L-92 and CP23 were subjected to two-dimensional electrophoresis, and major spots observed in the former were compared to the corresponding spots in the latter. To understand the effects of key components of L-92 on its adhesion to Caco-2 cells, 18 spots with stronger signals in L-92 than those in CP23 were identified by a MALDI-TOF/TOF of Ultraflex analysis. Among the identified proteins of L-92, surface-layer protein A (SlpA) was considered strongly involved in adhesion in the eight L. acidophilus strains. To study the importance of SlpA in the adhesion of L. acidophilus, the amounts of SlpA proteins in LiCl extracts of the eight strains were compared by SDSpolyacrylamide gel electrophoresis. As a result, the adhesive abilities of L. acidophilus strains to Caco-2 cells correlated closely to the amount of SlpA in the cells and the productivity of IL-12, an inflammatory cytokine, in all eight strains. These results strongly suggested that SlpA in L. acidophilus might play a key role in its attachment to Caco-2 cells and in the release of IL-12 from dendritic cells.

  2. More Protection of Lactobacillus acidophilus Than Bifidobacterium bifidum Probiotics on Azoxymethane-Induced Mouse Colon Cancer.

    PubMed

    Agah, Shahram; Alizadeh, Ali Mohammad; Mosavi, Maryam; Ranji, Peyman; Khavari-Daneshvar, Hossein; Ghasemian, Farnaz; Bahmani, Sahar; Tavassoli, Abbas

    2018-04-22

    Based on the ability of the probiotics in the gut microflora modification, they can have the beneficial effects on diseases in the short and/or the long term. In previous study, we revealed that unlike Bifidobacterium bifidum, the amount of Lactobacillus acidophilus remained almost unchanged in mice gut microflora in the long term, indicating more stability of L. acidophilus than B. bifidum which can be used to prevent some incurable diseases such as cancer. Thirty-eight male BALB/c mice were divided into four groups, control, azoxymethane (AOM), L. acidophilus, and B. bifidum probiotics, to evaluate the protective effects of the probiotics on AOM-induced mouse colon cancer. Except for the control group, the rest of the animals were weekly given AOM (15 mg/kg, s.c) in three consecutive weeks. Colon lesion incidence was 74% in the AOM group in comparison with the control (0%) (P < 0.05). The lesions were varied from mild to severe dysplasia and colonic adenocarcinoma. Administration of the probiotics inhibited the incidence of colonic lesions by about 57% in L. acidophilus (P < 0.05) and 27% in B. bifidum (P > 0.05) compared to the AOM group. The serum levels of CEA and CA19-9 tumor markers were significantly decreased in L. acidophilus in comparison with the AOM group (P < 0.05). Moreover, the serum levels of IFN-γ and IL-10 and the number of CD4 + and CD8 + cells were significantly increased in L. acidophilus compared to AOM (P < 0.05). Our study highlighted the more potential effects of L. acidophilus probiotic than B. bifidum on mouse colon cancer.

  3. Influence of Lactobacillus acidophilus and Lactobacillus plantarum on wound healing in male Wistar rats - an experimental study.

    PubMed

    Gudadappanavar, Anupama M; Hombal, Prashant R; Timashetti, Somling S; Javali, S B

    2017-01-01

    Probiotics have been documented with various pleotropic effects other than improving general gut health, but the potential benefits of strain-specific Lactobacillus on wound healing are unknown. Hence, the objective of the study is to evaluate and compare the wound healing property of Lactobacillus acidophilus and Lactobacillus plantarum on various wound models in male Wistar rats. Excision wound, resutured incision wound, and dead space wounds were inflicted under light thiopentone anesthesia in male Wistar rats ( n = 6, in each group). The rats received one of the Lactobacillus orally as per their weight for a period of 10 days in resutured incision (assessed by wound breaking strength) and dead space wounds (granuloma dry weight, histopathology of granulation tissue, and biochemical hydroxyproline estimation), whereas in excision wounds, treatment was monitored by planimetry. Data were expressed as mean ± standard error of mean and analyzed by ANOVA followed by Tukey's multiple post hoc test. P < 0.05 was considered as statistically significant. L. acidophilus showed a significant difference ( P < 0.05) in all the three models, namely, enhanced wound contraction and decreased days for complete epithelization in excision wound; increased breaking strength in resutured incision wound; increased granuloma dry weight and cellular infiltration in granulation tissue with marked increase in collagen content indicating wound healing. The study suggests that the wound healing activity of L. acidophilus if could be extrapolated to clinical situations may decrease dosage and duration of treatment and can be a potential adjuvant to reduce hospitalization with efficient recovery after injury and sustained good health.

  4. Effects of Lactobacillus acidophilus on the growth performance and intestinal health of broilers challenged with Clostridium perfringens.

    PubMed

    Li, Zhui; Wang, Weiwei; Liu, Dan; Guo, Yuming

    2018-01-01

    Clostridium perfringens is the main etiological agent of necrotic enteritis. Lactobacilli show beneficial effects on intestinal health in infectious disease, but the protective functions of lactobacilli in C. perfringens -infected chickens are scarcely described. This study examined the effects of Lactobacillus acidophilus ( L. acidophilus ) on the growth performance and intestinal health of broiler chickens challenged with Clostridium perfringens ( C. perfringens ) over a 28-day period. Using a 2 × 2 factorial arrangement of treatments, a total of 308 1-day-old male Arbor Acres broiler chicks were included to investigate the effects of Lactobacillus acidophilus ( L. acidophilus ) on the growth performance and intestinal health of broiler chickens challenged with Clostridium perfringens ( C. perfringens ) during a 28-day trial. During infection (d 14-21), C. perfringens challenge decreased the average daily gain ( P  <  0.05), and increased feed conversion ratio and the mortality rate ( P  <  0.05). However, dietary supplementation with L. acidophilus increased the body weight of C. perfringens -infected broilers on d 21 ( P  <  0.05), and tended to decrease the mortality ( P  = 0.061). C. perfringens challenge decreased the villus height ( P  <  0.05), the ratio of villus height to crypt depth ( P  <  0.05) and OCLN (occludin) mRNA expression ( P  <  0.05), and increased the pro-inflammatory cytokine expression in the spleen and jejunum, the intestinal populations of C. perfringens and Escherichia ( P  < 0.05), and the serum content of endotoxin ( P  < 0.05), regardless of L. acidophilus supplementation. In contrast, dietary L. acidophilus reducedthe intestinal lesion score of challenged broilers ( P  < 0.05), the mRNA expression of pro-inflammatory cytokines, ileal populations of Escherichia and serum endotoxin content ( P  < 0.05), but increased the intestinal Lactobacillus populations ( P  < 0

  5. 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

  6. Helicobacter pylori VacA Suppresses Lactobacillus acidophilus-Induced Interferon Beta Signaling in Macrophages via Alterations in the Endocytic Pathway

    PubMed Central

    Weiss, Gudrun; Forster, Sam; Irving, Aaron; Tate, Michelle; Ferrero, Richard L.; Hertzog, Paul; Frøkiær, Hanne; Kaparakis-Liaskos, Maria

    2013-01-01

    ABSTRACT Helicobacter pylori causes chronic gastritis and avoids elimination by the immune system of the infected host. The commensal bacterium Lactobacillus acidophilus has been suggested to exert beneficial effects as a supplement during H. pylori eradication therapy. In the present study, we applied whole-genome microarray analysis to compare the immune responses induced in murine bone marrow-derived macrophages (BMDMs) stimulated with L. acidophilus, H. pylori, or both bacteria in combination. While L. acidophilus induced a Th1-polarizing response characterized by high expression of interferon beta (IFN-β) and interleukin 12 (IL-12), H. pylori strongly induced the innate cytokines IL-1β and IL-1α. In BMDMs prestimulated with L. acidophilus, H. pylori blocked the expression of L. acidophilus-induced IFN-β and IL-12 and suppressed the expression of key regulators of the Rho, Rac, and Cdc42 GTPases. The inhibition of L. acidophilus-induced IFN-β was independent of H. pylori viability and the virulence factor CagPAI; however, a vacuolating cytotoxin (vacA) mutant was unable to block IFN-β. Confocal microscopy demonstrated that the addition of H. pylori to L. acidophilus-stimulated BMDMs redirects intracellular processing, leading to an accumulation of L. acidophilus in the endosomal and lysosomal compartments. Thus, our findings indicate that H. pylori inhibits the development of a strong Th1-polarizing response in BMDMs stimulated with L. acidophilus by blocking the production of IFN-β in a VacA-dependent manner. We suggest that this abrogation is caused by a redirection of the endocytotic pathway in the processing of L. acidophilus. PMID:23760466

  7. PP-12 EFFICACY OF A STANDARDIZED EXTRACT OF MATRICARIAE CHAMOMILLA L., MELISSA OFFICINALIS L. AND TYNDALLIZED LACTOBACILLUS ACIDOPHILUS (H122) COMPARED WITH LACTOBACILLUS REUTERI (DSM 17938) AND WITH SIMETHICONE FOR THE TREATMENT OF INFANTILE COLIC.

    PubMed

    Ummarino, Dario; Martinelli, Massimo; Giugliano, Francesca Paola; Tortora, Clelia; Valenti, Simona; Pidone, Caterina; De Giovanni, Donatella; Rutigliano, Irene; Campanozzi, Angelo; Romano, Claudio; Miele, Erasmo; Staiano, Annamaria

    2015-10-01

    Infant colic is a prevalent physiological event of healthy children under 3 months of age, which can disrupt the child's home environment. Despite its benign natural history, sometimes it requires a therapeutic approach. There is limited evidence supporting the use of complementary and alternative treatments. The aim of this study was to investigate the effectiveness of a mixture of standardized extract of matricariae Chamomilla L., Melissa Officinalis L. and tyndallized Lactobacillus Acidophilus (H122) compared with Lactobacillus Reuteri (DSM 17938) and with simethicone for the treatment of infantile colic. A multicenter prospective, randomized comparative study was conducted involving 133 infants with colic, according to Rome III criteria, who were assigned at random to receive Chamomilla L., Melissa Officinalis L. and tyndallized L. Acidophilus (H122) (Group A; n = 45), L reuteri DSM 17938 (10 CFU) (Group B; n = 45) or simethicone (Group C; n = 43). Treatment was given to subjects for 21 days and they were followed for 4 weeks. Treatment success was assessed at the end of study period. Daily crying and fussing times were recorded in a structured diary, and maternal questionnaires were completed to monitor changes in infant colic symptoms and adverse events. Treatment success was observed in 30 out of 45 patients (66.7%) of infants in Group A, while it was seen in 31 out of 45 patients (68.9%) of Group B and in 19 out of 43 (44.2%) of subjects of Group C (χ: 6.8; p = 0.03). Mean daily crying time was more significantly reduced (from 211.3 ± 40 min/day to 69.6 ± 59 min/day) in the Group A and in Group B (from 201.6 ± 32.5 min/day to 58.1 ± 48.9 min/day) when compared with Group C (from 199.5 ± 32 min/day to 106 ± 56.5 min/day) (p < 0.01, p = 0.006 respectively). No significant difference was observed comparing Group A and B (p = 0.4). No adverse events were reported in any group. This study suggests

  8. Lactobacillus acidophilus ATCC 4356 inhibits biofilm formation by C. albicans and attenuates the experimental candidiasis in Galleria mellonella.

    PubMed

    Vilela, Simone F G; Barbosa, Júnia O; Rossoni, Rodnei D; Santos, Jéssica D; Prata, Marcia C A; Anbinder, Ana Lia; Jorge, Antonio O C; Junqueira, Juliana C

    2015-01-01

    Probiotic strains of Lactobacillus have been studied for their inhibitory effects on Candida albicans. However, few studies have investigated the effect of these strains on biofilm formation, filamentation and C. albicans infection. The objective of this study was to evaluate the influence of Lactobacillus acidophilus ATCC 4356 on C. albicans ATCC 18804 using in vitro and in vivo models. In vitro analysis evaluated the effects of L. acidophilus on the biofilm formation and on the capacity of C. albicans filamentation. For in vivo study, Galleria mellonella was used as an infection model to evaluate the effects of L. acidophilus on candidiasis by survival analysis, quantification of C. albicans CFU/mL, and histological analysis. The direct effects of L. acidophilus cells on C. albicans, as well as the indirect effects using only a Lactobacillus culture filtrate, were evaluated in both tests. The in vitro results showed that both L. acidophilus cells and filtrate were able to inhibit C. albicans biofilm formation and filamentation. In the in vivo study, injection of L. acidophilus into G. mellonella larvae infected with C. albicans increased the survival of these animals. Furthermore, the number of C. albicans CFU/mL recovered from the larval hemolymph was lower in the group inoculated with L. acidophilus compared to the control group. In conclusion, L. acidophilus ATCC 4356 inhibited in vitro biofilm formation by C. albicans and protected G. mellonella against experimental candidiasis in vivo.

  9. Lactobacillus acidophilus ATCC 4356 inhibits biofilm formation by C. albicans and attenuates the experimental candidiasis in Galleria mellonella

    PubMed Central

    Vilela, Simone FG; Barbosa, Júnia O; Rossoni, Rodnei D; Santos, Jéssica D; Prata, Marcia CA; Anbinder, Ana Lia; Jorge, Antonio OC; Junqueira, Juliana C

    2015-01-01

    Probiotic strains of Lactobacillus have been studied for their inhibitory effects on Candida albicans. However, few studies have investigated the effect of these strains on biofilm formation, filamentation and C. albicans infection. The objective of this study was to evaluate the influence of Lactobacillus acidophilus ATCC 4356 on C. albicans ATCC 18804 using in vitro and in vivo models. In vitro analysis evaluated the effects of L. acidophilus on the biofilm formation and on the capacity of C. albicans filamentation. For in vivo study, Galleria mellonella was used as an infection model to evaluate the effects of L. acidophilus on candidiasis by survival analysis, quantification of C. albicans CFU/mL, and histological analysis. The direct effects of L. acidophilus cells on C. albicans, as well as the indirect effects using only a Lactobacillus culture filtrate, were evaluated in both tests. The in vitro results showed that both L. acidophilus cells and filtrate were able to inhibit C. albicans biofilm formation and filamentation. In the in vivo study, injection of L. acidophilus into G. mellonella larvae infected with C. albicans increased the survival of these animals. Furthermore, the number of C. albicans CFU/mL recovered from the larval hemolymph was lower in the group inoculated with L. acidophilus compared to the control group. In conclusion, L. acidophilus ATCC 4356 inhibited in vitro biofilm formation by C. albicans and protected G. mellonella against experimental candidiasis in vivo. PMID:25654408

  10. Effects of Lactobacillus rhamnosus and Lactobacillus acidophilus on bacterial vaginal pathogens.

    PubMed

    Bertuccini, Lucia; Russo, Rosario; Iosi, Francesca; Superti, Fabiana

    2017-06-01

    The human vagina is colonized by a variety of microbes. Lactobacilli are the most common, mainly in healthy women; however, the microbiota composition can change rapidly, leading to infection or to a state in which potential pathogenic microorganisms co-exist with other commensals. In premenopausal women, urogenital infections, such as bacterial vaginosis and aerobic vaginitis, remain an important health problem. Treatment of these infections involves different kind of antibiotics; however, the recurrence rate remains high, and it must be also underlined that antibiotics are unable to spontaneously restore normal flora characterized by an abundant community of Lactobacilli. The main limitation is the inability to offer a long-term defensive barrier, thus facilitating relapses and recurrences. We report here the antimicrobial activities of two commercially existing Lactobacillus strains, Lactobacillus rhamnosus HN001 and Lactobacillus acidophilus GLA-14 strains and their combination (Respecta® probiotic blend) against four different pathogens responsible for both bacterial vaginosis ( Gardenerella vaginalis and Atopobium vaginae) and aerobic vaginitis ( Staphylococcus aureus and Escherichia coli) by co-culturing assay. The probiotic combination, even if resulting in a different microbicidal activity against the different strains tested, demonstrated the efficacy of combined Lactobacillus strain treatment.

  11. Effects of Lactobacillus rhamnosus and Lactobacillus acidophilus on bacterial vaginal pathogens

    PubMed Central

    Bertuccini, Lucia; Russo, Rosario; Iosi, Francesca; Superti, Fabiana

    2017-01-01

    The human vagina is colonized by a variety of microbes. Lactobacilli are the most common, mainly in healthy women; however, the microbiota composition can change rapidly, leading to infection or to a state in which potential pathogenic microorganisms co-exist with other commensals. In premenopausal women, urogenital infections, such as bacterial vaginosis and aerobic vaginitis, remain an important health problem. Treatment of these infections involves different kind of antibiotics; however, the recurrence rate remains high, and it must be also underlined that antibiotics are unable to spontaneously restore normal flora characterized by an abundant community of Lactobacilli. The main limitation is the inability to offer a long-term defensive barrier, thus facilitating relapses and recurrences. We report here the antimicrobial activities of two commercially existing Lactobacillus strains, Lactobacillus rhamnosus HN001 and Lactobacillus acidophilus GLA-14 strains and their combination (Respecta® probiotic blend) against four different pathogens responsible for both bacterial vaginosis (Gardenerella vaginalis and Atopobium vaginae) and aerobic vaginitis (Staphylococcus aureus and Escherichia coli) by co-culturing assay. The probiotic combination, even if resulting in a different microbicidal activity against the different strains tested, demonstrated the efficacy of combined Lactobacillus strain treatment. PMID:28580872

  12. 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. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  13. Plasma membrane Toll-like receptor activation increases bacterial uptake but abrogates endosomal Lactobacillus acidophilus induction of interferon-β.

    PubMed

    Boye, Louise; Welsby, Iain; Lund, Lisbeth Drozd; Goriely, Stanislas; Frøkiaer, Hanne

    2016-11-01

    Lactobacillus acidophilus induces a potent interferon-β (IFN-β) response in dendritic cells (DCs) by a Toll-like receptor 2 (TLR2) -dependent mechanism, in turn leading to strong interleukin-12 (IL-12) production. In the present study, we investigated the involvement of different types of endocytosis in the L. acidophilus-induced IFN-β and IL-12 responses and how TLR2 or TLR4 ligation by lipopolysaccharide and Pam3/4CSK4 influenced endocytosis of L. acidophilus and the induced IFN-β and IL-12 production. Lactobacillus acidophilus was endocytosed by constitutive macropinocytosis taking place in the immature cells as well as by spleen tyrosine kinase (Syk) -dependent phagocytosis but without involvement of plasma membrane TLR2. Stimulation with TLR2 or TLR4 ligands increased macropinocytosis in a Syk-independent manner. As a consequence, incubation of DCs with TLR ligands before incubation with L. acidophilus enhanced the uptake of the bacteria. However, in these experimental conditions, induction of IFN-β and IL-12 was strongly inhibited. As L. acidophilus-induced IFN-β depends on endocytosis and endosomal degradation before signalling and as TLR stimulation from the plasma membrane leading to increased macropinocytosis abrogates IFN-β induction we conclude that plasma membrane TLR stimulation leading to increased macropinocytosis decreases endosomal induction of IFN-β and speculate that this is due to competition between compartments for molecules involved in the signal pathways. In summary, endosomal signalling by L. acidophilus that leads to IFN-β and IL-12 production is inhibited by TLR stimulation from the plasma membrane. © 2016 John Wiley & Sons Ltd.

  14. Development and use of tuf gene-based primers for the multiplex PCR detection of Lactobacillus acidophilus, Lactobacillus casei group, Lactobacillus delbrueckii, and Bifidobacterium longum in commercial dairy products.

    PubMed

    Sheu, Sen-Je; Hwang, Wen-zhe; Chen, Hsin-Chih; Chiang, Yu-Cheng; Tsen, Hau-Yang

    2009-01-01

    PCR primers specific for the detection of Lactobacillus acidophilus, Lactobacillus casei group, Lactobacillus delbrueckii, and Bifidobacterium longum were designed based on the elongation factor Tu gene (tuf). The specificity of these four primer sets were confirmed by PCR with 88 bacterial strains of Lactobacillus, Enterococcus, Bifidobacterium, and other bacterial species. Results indicated that these primer sets generated predicted PCR products of 397, 230, 202, and 161 bp for L. acidophilus, L. delbrueckii, L. casei group, and B. longum, respectively. Bacterial species other than the target organisms tested did not generate false-positive results. When these four primer sets were combined for the simultaneous detection of the lactic acid bacteria (LAB) in fermented milk products including yogurt, the LAB species listed on the labels of these products could be identified without the preenrichment step. The identification limit for each LAB strain with this multiplex PCR method was N X 10(3) CFU/ml in milk samples. The results of our multiplex PCR method were confirmed by PCR assay using primers based on the 16S rDNA or the 16S-23S intergenic spacer region and by biochemical tests using the API 50 CHL kit. When this multiplex PCR method was used with the determination of counts of total viable LAB and bifidobacteria, the quality of commercial fermented milk products could be assured.

  15. Mucin- and carbohydrate-stimulated adhesion and subproteome changes of the probiotic bacterium Lactobacillus acidophilus NCFM.

    PubMed

    Celebioglu, Hasan Ufuk; Olesen, Sita Vaag; Prehn, Kennie; Lahtinen, Sampo J; Brix, Susanne; Abou Hachem, Maher; Svensson, Birte

    2017-06-23

    Adhesion to intestinal mucosa is a crucial property for probiotic bacteria. Adhesion is thought to increase host-bacterial interactions, thus potentially enabling health benefits to the host. Molecular events connected with adhesion and surface proteome changes were investigated for the probiotic Lactobacillus acidophilus NCFM cultured with established or emerging prebiotic carbohydrates as carbon source and in the presence of mucin, the glycoprotein of the epithelial mucus layer. Variation in adhesion to HT29-cells and mucin was associated with carbon source and mucin-induced subproteome abundancy differences. Specifically, while growth on fructooligosaccharides (FOS) only stimulated adhesion to intestinal HT-29 cells, cellobiose and polydextrose in addition increased adhesion to mucin. Adhesion to HT-29 cells increased by about 2-fold for bacteria grown on mucin-supplemented glucose. Comparative 2DE-MS surface proteome analysis showed different proteins in energy metabolism appearing on the surface, suggesting they exert moonlighting functions. Mucin-supplemented bacteria had relative abundance of pyruvate kinase and fructose-bisphosphate aldolase increased by about 2-fold while six spots with 3.2-2.1 fold reduced relative abundance comprised elongation factor G, phosphoglycerate kinase, BipAEFTU family GTP-binding protein, ribonucleoside triphosphate reductase, adenylosuccinate synthetase, 30S ribosomal protein S1, and manganese-dependent inorganic pyrophosphatase. Surface proteome of cellobiose- compared to glucose-grown L. acidophilus NCFM had phosphate starvation inducible protein stress-related, thermostable pullulanase, and elongation factor G increasing 4.4-2.4 fold, while GAPDH, elongation factor Ts, and pyruvate kinase were reduced by 2.0-1.5 fold in relative abundance. Addition of recombinant L. acidophilus NCFM elongation factor G and pyruvate kinase to a coated mucin layer significantly suppressed subsequent adhesion of the bacterium. Human diet is

  16. Identification of extracellular surface-layer associated proteins in Lactobacillus acidophilus NCFM

    PubMed Central

    Johnson, Brant; Selle, Kurt; O’Flaherty, Sarah; Goh, Yong Jun

    2013-01-01

    Bacterial surface (S-) layers are crystalline arrays of self-assembling, proteinaceous subunits called S-layer proteins (Slps), with molecular masses ranging from 40 to 200 kDa. The S-layer-forming bacterium Lactobacillus acidophilus NCFM expresses three major Slps: SlpA (46 kDa), SlpB (47 kDa) and SlpX (51 kDa). SlpA has a demonstrated role in adhesion to Caco-2 intestinal epithelial cells in vitro, and has been shown to modulate dendritic cell (DC) and T-cell functionalities with murine DCs. In this study, a modification of a standard lithium chloride S-layer extraction revealed 37 proteins were solubilized from the S-layer wash fraction. Of these, 30 have predicted cleavage sites for secretion, 24 are predicted to be extracellular, six are lipid-anchored, three have N-terminal hydrophobic membrane spanning regions and four are intracellular, potentially moonlighting proteins. Some of these proteins, designated S-layer associated proteins (SLAPs), may be loosely associated with or embedded within the bacterial S-layer complex. Lba-1029, a putative SLAP gene, was deleted from the chromosome of L. acidophilus. Phenotypic characterization of the deletion mutant demonstrated that the SLAP LBA1029 contributes to a pro-inflammatory TNF-α response from murine DCs. This study identified extracellular proteins and putative SLAPs of L. acidophilus NCFM using LC-MS/MS. SLAPs appear to impart important surface display features and immunological properties to microbes that are coated by S-layers. PMID:24002751

  17. [Expression, purification and activity analysis of GGDEF and EAL domain-containing proteins from Lactobacillus acidophilus].

    PubMed

    He, Jia-Hui; Sun, Jie-Li; Yan, Wen-Juan; Wang, Fang

    2017-05-20

    To identify the functions of the proteins containing the GGDEF or EAL domain in Lactobacillus acidophilus for investigation of the regulatory mechanism of c-di-GMP in this strain. The DNA fragments of NH13_07045-GGDEF, NH13_07050 and NH13_07055 from Lactobacillus acidophilus ATCC4356 were amplified by PCR and cloned into the expression vector pMAL-His-c2. After sequencing, the recombinant plasmids were transformed into competent Escherichia coli cells, which were induced by IPTG to express the recombinant proteins fused with maltose binding protein (MBP). The fusion proteins were purified using amylose resin column for diguanylate cyclase (DGC) or phosphodiesterase (PDE) activity assays in vitro followed by analysis with high-performance liquid chromatography (HPLC). The target DNA fragments were obtained by PCR, and their sequences were all identical to that in GenBank. The purified and concentrated fusion proteins, which were identified by SDS-PAGE and Western blotting, had relative molecular masses of 59 kD, 67 kD and 72 kD. HPLC analysis showed no DGC activity in NH13_07045-GGDEF, while PDE activity was found in NH13_07050 but not in NH13_07055. We obtained the protein encoded by NH13_07050 that possesses PDE activity in vitro. This protein may facilitate the evaluation of the regulatory function of c-di-GMP in Lactobacillus acidophilus.

  18. Response of Lactobacillus acidophilus ATCC 4356 to low-shear modeled microgravity

    NASA Astrophysics Data System (ADS)

    Castro-Wallace, Sarah; Stahl, Sarah; Voorhies, Alexander; Lorenzi, Hernan; Douglas, Grace L.

    2017-10-01

    The introduction of 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 gene expression of probiotic bacteria must be investigated to confirm that benefits of selected strains will still be conveyed under microgravity conditions. The goal of this study was to evaluate the characteristics of the probiotic bacteria Lactobacillus acidophilus ATCC 4356 in a microgravity analog environment. L. acidophilus was cultured anaerobically under modeled microgravity conditions and assessed for differences in growth, survival through stress challenge, and gene expression compared to control cultures. No significant differences were observed between the modeled microgravity and control grown L. acidophilus, suggesting that this strain will behave similarly in spaceflight.

  19. Predominant genera of fecal microbiota in children with atopic dermatitis are not altered by intake of probiotic bacteria Lactobacillus acidophilus NCFM and Bifidobacterium animalis subsp. lactis Bi-07.

    PubMed

    Larsen, Nadja; Vogensen, Finn K; Gøbel, Rikke; Michaelsen, Kim F; Abu Al-Soud, Waleed; Sørensen, Søren J; Hansen, Lars H; Jakobsen, Mogens

    2011-03-01

    The effect of probiotic bacteria Lactobacillus acidophilus NCFM and Bifidobacterium lactis Bi-07 on the composition of the Lactobacillus group, Bifidobacterium and the total bacterial population in feces from young children with atopic dermatitis was investigated. The study included 50 children randomized to intake of one of the probiotic strain or placebo. Microbial composition was characterized by denaturing gradient gel electrophoresis, quantitative PCR and, in a subset of subjects, by pyrosequencing of the 16S rRNA gene. The core population of the Lactobacillus group was identified as Lactobacillus gasseri, Lactobacillus fermentum, Lactobacillus oris, Leuconostoc mesenteroides, while the bifidobacterial community included Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium longum and Bifidobacterium catenulatum. The fecal numbers of L. acidophilus and B. lactis increased significantly after intervention, indicating survival of the ingested bacteria. The levels of Bifidobacterium correlated positively (P=0.03), while the levels of the Lactobacillus group negatively (P=0.01) with improvement of atopic eczema evaluated by the Severity Scoring of Atopic Dermatitis index. This correlation was observed across the whole study cohort and not attributed to the probiotic intake. The main conclusion of the study is that administration of L. acidophilus NCFM and B. lactis Bi-07 does not affect the composition and diversity of the main bacterial populations in feces. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  20. Lactobacillus acidophilus CL1285, Lactobacillus casei LBC80R, and Lactobacillus rhamnosus CLR2 (Bio-K+): Characterization, Manufacture, Mechanisms of Action, and Quality Control of a Specific Probiotic Combination for Primary Prevention of Clostridium difficile Infection.

    PubMed

    Auclair, Julie; Frappier, Martin; Millette, Mathieu

    2015-05-15

    A specific probiotic formulation composed of Lactobacillus acidophilus CL1285, Lactobacillus casei LBC80R, and Lactobacillus rhamnosus CLR2 (Bio-K+) has been marketed in North America since 1996. The strains and the commercial products have been evaluated for safety, identity, gastrointestinal survival, and stability throughout shelf life. The capacity of both the fermented beverages and the capsules to reduce incidences of antibiotic-associated diarrhea and Clostridium difficile infection (CDI) has been demonstrated in human clinical trials. Individual strains and the finished products have shown antimicrobial activity against C. difficile and toxin A/B neutralization capacity in vitro. The use of this specific probiotic formulation as part of a bundle of preventive measures to control CDI in healthcare settings is discussed. © The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  1. Effects of Lactobacillus acidophilus dietary supplementation on the performance, intestinal barrier function, rectal microflora and serum immune function in weaned piglets challenged with Escherichia coli lipopolysaccharide.

    PubMed

    Qiao, Jiayun; Li, Haihua; Wang, Zhixiang; Wang, Wenjie

    2015-04-01

    This study was conducted with a lipopolysaccharide (LPS)-challenged piglet model to determine the effects of diets containing Lactobacillus acidophilus on the performance, intestinal barrier function, rectal microflora and serum immune function. A total of 150 piglets (initial body weight (BW) 7.53 ± 0.21 kg) were allotted to one of the following diets, including a basal diet, a basal diet supplemented with 250 mg/kg Flavomycin, or basal diet plus 0.05, 0.1 or 0.2 % L. acidophilus. On day 28 of the trial, the pigs were given an intraperitoneal injection of LPS (200 μg/kg body weight) followed by blood collection 3 h later. Diets with either antibiotics, 0.1 or 0.2 % Lactobacillus increased (P < 0.05) the final BW and decreased (P < 0.05) feed gain ratio (F/G) compared with the control group. Pigs fed diets containing antibiotic or Lactobacillus had greater average daily gain (ADG) (P < 0.05) than pigs fed the control diet. The rectal content Lactobacillus counts for pigs fed diet containing Lactobacillus were significant higher (P < 0.01) than those fed antibiotic or control diet. Feeding the Lactobacillus diets decreased the Escherichia coli counts of rectal content (P < 0.01). Pigs fed diets containing 0.1 or 0.2 % Lactobacillus decreased serum DAO activity (P < 0.05) compared with pigs fed the control diet. Serum IL-10 concentration was enhanced in pigs fed the diet with Lactobacillus compared to pigs fed the control diet and antibiotic diet. Feeding a diet with Lactobacillus reduced (P < 0.05) IFN-γ concentration compared to the control diet. Inclusion of Lactobacillus in diets fed to pigs reduced TNF-α concentration compared with pigs fed no Lactobacillus (P < 0.05). These results indicate that feeding with L. acidophilus improved growth performance and protected against LPS-induced inflammatory status.

  2. Effect of salt on cell viability and membrane integrity of Lactobacillus acidophilus, Lactobacillus casei and Bifidobacterium longum as observed by flow cytometry.

    PubMed

    Gandhi, Akanksha; Shah, Nagendra P

    2015-08-01

    The aim of the current study was to investigate the effect of varying sodium chloride concentrations (0-5%) on viability and membrane integrity of three probiotic bacteria, Lactobacillus acidophilus, Lactobacillus casei and Bifidobacterium longum, using conventional technique and flow cytometry. Double staining of cells by carboxyfluorescein diacetate (cFDA) and propidium iodide (PI) enabled to evaluate the effect of NaCl on cell esterase activity and membrane integrity. Observations from conventional culture technique were compared with findings from flow cytometric analysis on the metabolic activities of the cells and a correlation was observed between culturability and dye extrusion ability of L. casei and B. longum. However, a certain population of L. acidophilus was viable as per the plate count method but its efflux activity was compromised. Esterase activity of most bacteria reduced significantly (P < 0.05) during one week storage at NaCl concentrations greater than 3.5%. The study revealed that L. casei was least affected by higher NaCl concentrations among the three probiotic bacteria, as opposed to B. longum where the cF extrusion performance was greatly reduced during 1 wk storage. The metabolic activity and salt resistance of L. casei was found to be highest among the bacteria studied. Crown Copyright © 2015. Published by Elsevier Ltd. All rights reserved.

  3. Lactobacillus acidophilus ATCC 4356 attenuates the atherosclerotic progression through modulation of oxidative stress and inflammatory process.

    PubMed

    Chen, Lihua; Liu, Wenen; Li, Yanming; Luo, San; Liu, Qingxia; Zhong, Yiming; Jian, Zijuan; Bao, Meihua

    2013-09-01

    The aim of this study was to investigate the effect of Lactobacillus (L.) acidophilus ATCC 4356 on the progression of atherosclerosis in Apoliprotein-E knockout (ApoE(-/-)) mice and the underlying mechanisms. Eight week-old ApoE(-/-) mice were treated with L. acidophilus ATCC 4356 daily for 12 weeks. The wild type (WT) mice or ApoE(-/-) mice in the vehicle group were treated with saline only. Body weights, serum lipid levels, aortic atherosclerotic lesions, and tissue oxidative and inflammatory statuses were examined among the groups. As compared to ApoE(-/-) mice in the vehicle group, ApoE(-/-) mice treated with L. acidophilus ATCC 4356 had no changes in body weights and serum lipid profiles, but showed decreased atherosclerotic lesion size in en face aorta. In comparison with WT mice, ApoE(-/-) mice in the vehicle group showed higher levels of serum malondialdehyde (MDA), oxidized low density lipoprotein (oxLDL) and tumor necrosis factor-alpha (TNF-α), but lower levels of interleukin-10 (IL-10) and superoxide dismutase (SOD) activities in serum. Administration of L. acidophilus ATCC 4356 could reverse these trends in a dose-dependent manner in ApoE(-/-) mice. Furthermore, ApoE(-/-) mice treated with L. acidophilus ATCC 4356 showed an inhibition of translocation of NF-κB p65 from cytoplasm to nucleus, suppression of degradation of aortic IκB-α, and improvements of gut microbiota distribution, as compared to ApoE(-/-) mice in the vehicle group. Our findings suggest that administration of L. acidophilus ATCC 4356 can attenuate the development of atherosclerotic lesions in ApoE(-/-) mice through reducing oxidative stress and inflammatory response. Copyright © 2013 Elsevier B.V. All rights reserved.

  4. Inhibition of Cronobacter sakazakii by Lactobacillus acidophilus n.v. Er2 317/402.

    PubMed

    Charchoghlyan, Haykuhi; Kwon, Heejun; Hwang, Dong-Ju; Lee, Jong Suk; Lee, Junsoo; Kim, Myunghee

    2016-10-31

    Lactobacillus acidophilus n.v. Er2 317/402 strain Narine is known as a health beneficial functional probiotic culture and supplementary source of nutrition for newborns. In this study, in vitro antimicrobial activities of Narine-lyophilized (Narine-L), Narine-heat treated (Narine-HT), and Narine crude cell-free extract (Narine-CCFE) were evaluated against pathogen Cronobacter sakazakii ( C. sakazakii ) in agar as well as in a reconstituted powdered infant formula (RPIF) model. Inhibition zones of 30 mg Narine-L and Narine-HT were both 150 U, whereas inhibition zone of 30 mg Narine-CCFE was 200 U. Narine-L (1 g) and Narine-HT (1 g) were added to 10 mL of artificially contaminated RPIF, respectively, containing 100 μL of C. sakazakii (1.62×10 8 colony forming unit (CFU)/mL). After treatment with Narine-L and Narine-HT for 3 h and 6 h at 37℃, less than ≤107 CFU/mL of C. sakazakii was detected in RPIF. Without Narine-L and Narine-HT treatment, the population of C. sakazakii increased up to 5.36×10 9 CFU/mL after 6 h. Examination by transmission electron microscopy confirmed C. sakazakii cells were damaged by Narine-CCFE. Thus, employing Narine culture as a natural and safe bio-preservative may protect infants from C. sakazakii .

  5. The effects of Lactobacillus Acidophilus fermentation products as an alternative to antibiotics

    USDA-ARS?s Scientific Manuscript database

    This experiment compared the effects of Lactobacillus acidophilus fermentation products (LAFP) to carbadox and copper sulfate on growth performance and complete blood counts (CBC). Eight hundred pigs were weaned at 24 d of age and utilized in a randomized block design (4 farrowing groups, blocked by...

  6. Lactobacillus acidophilus attenuates Salmonella-induced intestinal inflammation via TGF-β signaling.

    PubMed

    Huang, I-Fei; Lin, I-Chun; Liu, Pei-Feng; Cheng, Ming-Fang; Liu, Yen-Chen; Hsieh, Yao-Dung; Chen, Jih-Jung; Chen, Chun-Lin; Chang, Hsueh-Wei; Shu, Chih-Wen

    2015-10-07

    Salmonella is a common intestinal pathogen that causes acute and chronic inflammatory response. Probiotics reduce inflammatory cytokine production and serve as beneficial commensal microorganisms in the human gastrointestinal tract. TGF-β (transforming growth factor β)/SMAD and NF-κB signaling play important roles in inflammation in intestinal cells. However, the involvement of the signaling in regulating inflammation between Salmonella and probiotics is not fully understood. L. acidophilus and prebiotic inulin were used to treat human intestinal Caco-2 cells prior to infection with Salmonella. The cells were harvested to examine the cytokines and MIR21 expression with immunoblotting and real-time PCR. NF-κB and SMAD3/4 reporter vectors were transfected into cells to monitor inflammation and TGF-β1 signaling, respectively. In this study, we showed that the probiotic L. acidophilus decreased Salmonella-induced NF-κB activation in human intestinal Caco-2 cells. Expression of the inflammatory cytokines, TNF-α and IL-8, in L. acidophilus-pretreated cells was also significantly lower than that in cells infected with Salmonella alone. Moreover, TGF-β1 and MIR21 expression was elevated in cells pretreated with L. acidophilus or synbiotic, a combination of inulin and L. acidophilus, compared to that in untreated cells or cells infected with S. typhimurium alone. By contrast, expression of SMAD7, a target of MIR21, was accordingly reduced in cells treated with L. acidophilus or synbiotics. Consistent with TGF-β1/MIR21 and SMAD7 expression, SMAD3/4 transcriptional activity was significantly higher in the cells treated with L. acidophilus or synbiotics. Furthermore, TGF-β1 antibody antagonized the SMAD3/4 and NF-κB transcriptional activity modulated by L. acidophilus in intestinal cells. Our results suggest that the TGF-β1/MIR21 signaling pathway may be involved in the suppressive effects of L. acidophilus on inflammation caused by S. typhimurium in intestinal

  7. 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. PMID:26761878

  8. The substitution of a traditional starter culture in mutton fermented sausages by Lactobacillus acidophilus and Bifidobacterium animalis.

    PubMed

    Holko, I; Hrabě, J; Šalaková, A; Rada, V

    2013-07-01

    Common starter cultures used in fermented mutton sausages were substituted by probiotic strains of Lactobacillus acidophilus CCDM 476 and Bifidobacterium animalis 241a. Technological properties of the traditional and the probiotic sausages were compared. The potential probiotic effect was evaluated by enumeration of bifidobacteria and lactobacilli in stool samples of 15 volunteers before and after a 14-day consumption period. The numbers of lactobacilli (10(7) cfu/g) and bifidobacteria (10(3) cfu/g) in the final product did not affect the technological properties. The use of L. acidophilus as a starter culture was found more beneficial than the use of B. animalis. Even after 60 days of storage, high counts of L. acidophilus (10(6) cfu/g) were detected; on the other hand, the counts of B. animalis were under the detection limit. Regarding sensory properties, the probiotic products showed better texture, and, curiously, a reduction of the typical smell of mutton. The numbers of lactobacilli in stool samples increased significantly after the consumption of the probiotic sausages. Copyright © 2013 Elsevier Ltd. All rights reserved.

  9. Lactobacillus acidophilus-Rutin Interplay Investigated by Proteomics.

    PubMed

    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.

  10. Dietary Lactobacillus acidophilus positively influences growth performance, gut morphology, and gut microbiology in rurally reared chickens.

    PubMed

    Forte, C; Manuali, E; Abbate, Y; Papa, P; Vieceli, L; Tentellini, M; Trabalza-Marinucci, M; Moscati, L

    2018-03-01

    In a market undergoing constant evolution, the production of chicken meat that consumers would perceive as "natural" and "animal friendly" is crucial. The use of probiotics in rurally reared chickens could represent a major opportunity to achieve mutual benefit for both the industry and consumers. A total of 264 male Kabir chicks were randomly distributed to one of 2 dietary treatments: the L group received a commercial feed supplemented with 2.0 g/100 kg of Lactobacillus acidophilus D2/CSL, while the C group received the same basal diet without the additive. To assess the effects of probiotic supplementation in the chickens' diet, productive performance was evaluated at d 21 and 42, whereas microbiological analyses of the intestinal content and intestinal histology and morphometry were performed at the end of the trial (d 42). At d 21 and 42, L birds showed better (P < 0.001) performance in terms of body weight, average daily gain, and feed conversion ratio. Enterococci, staphylococci, and Escherichia coli populations were not influenced by dietary treatment. On the contrary, Lactobacillus population increased (P = 0.032) in the L group. Furthermore, a tendency (P = 0.069) was observed for the coliforms to be influenced by diet, with lower values in the L group in comparison to the C group. Histological techniques revealed that the number of goblet cell containing neutral mucins was lower in the C group. Morphometric evaluations demonstrated that the probiotic supplementation increased the height of the mucosal layer by improving (P = 0.040) villus height, while crypt depth was unaffected. In conclusion, the results obtained in this study demonstrate that it is possible to use Lactobacillus acidophilus D2/CSL (CECT 4529) in rurally reared chicken breeds with positive effects on performance and gut health.

  11. Dietary Lactobacillus acidophilus positively influences growth performance, gut morphology, and gut microbiology in rurally reared chickens

    PubMed Central

    Forte, C; Manuali, E; Abbate, Y; Papa, P; Vieceli, L; Tentellini, M; Trabalza-Marinucci, M; Moscati, L

    2018-01-01

    Abstract In a market undergoing constant evolution, the production of chicken meat that consumers would perceive as “natural” and “animal friendly” is crucial. The use of probiotics in rurally reared chickens could represent a major opportunity to achieve mutual benefit for both the industry and consumers. A total of 264 male Kabir chicks were randomly distributed to one of 2 dietary treatments: the L group received a commercial feed supplemented with 2.0 g/100 kg of Lactobacillus acidophilus D2/CSL, while the C group received the same basal diet without the additive. To assess the effects of probiotic supplementation in the chickens’ diet, productive performance was evaluated at d 21 and 42, whereas microbiological analyses of the intestinal content and intestinal histology and morphometry were performed at the end of the trial (d 42). At d 21 and 42, L birds showed better (P < 0.001) performance in terms of body weight, average daily gain, and feed conversion ratio. Enterococci, staphylococci, and Escherichia coli populations were not influenced by dietary treatment. On the contrary, Lactobacillus population increased (P = 0.032) in the L group. Furthermore, a tendency (P = 0.069) was observed for the coliforms to be influenced by diet, with lower values in the L group in comparison to the C group. Histological techniques revealed that the number of goblet cell containing neutral mucins was lower in the C group. Morphometric evaluations demonstrated that the probiotic supplementation increased the height of the mucosal layer by improving (P = 0.040) villus height, while crypt depth was unaffected. In conclusion, the results obtained in this study demonstrate that it is possible to use Lactobacillus acidophilus D2/CSL (CECT 4529) in rurally reared chicken breeds with positive effects on performance and gut health. PMID:29294082

  12. Cell growth and proteolytic activity of Lactobacillus acidophilus, Lactobacillus helveticus, Lactobacillus delbrueckii ssp. bulgaricus, and Streptococcus thermophilus in milk as affected by supplementation with peptide fractions.

    PubMed

    Gandhi, Akanksha; Shah, Nagendra P

    2014-12-01

    The present investigation examined the effects of supplementation of milk peptide fractions produced by enzymatic hydrolysis on the fermentation of reconstituted skim milk (RSM). Changes in pH, cell growth, proteolytic activity, and angiotensin-converting enzyme (ACE)-inhibitory activity were monitored during fermentation of RSM by pure cultures of Lactobacillus acidophilus, Lactobacillus helveticus, Lactobacillus delbrueckii ssp. bulgaricus, and Streptococcus thermophilus. The study showed that supplementation with peptide fractions of different molecular weights did not significantly affect the bacterial growth in RSM. All bacteria showed an increased proteolytic activity in RSM supplemented with large peptides (>10 kDa), and L. helveticus in general exhibited the highest proteolytic activity among the bacteria studied. The ACE-inhibitory activity was observed to be the maximum in RSM supplemented with larger peptides (>10 kDa) for all bacteria. The results suggest that proteolysis by bacteria leads to increased production of ACE-inhibitory peptides compared to the supplemented peptides produced by enzymatic hydrolysis.

  13. Lactobacillus acidophilus Metabolizes Dietary Plant Glucosides and Externalizes Their Bioactive Phytochemicals.

    PubMed

    Theilmann, Mia C; Goh, Yong Jun; Nielsen, Kristian Fog; Klaenhammer, Todd R; Barrangou, Rodolphe; Abou Hachem, Maher

    2017-11-21

    Therapeutically active glycosylated phytochemicals are ubiquitous in the human diet. The human gut microbiota (HGM) modulates the bioactivities of these compounds, which consequently affect host physiology and microbiota composition. Despite a significant impact on human health, the key players and the underpinning mechanisms of this interplay remain uncharacterized. Here, we demonstrate the growth of Lactobacillus acidophilus on mono- and diglucosyl dietary plant glycosides (PGs) possessing small aromatic aglycones. Transcriptional analysis revealed the upregulation of host interaction genes and identified two loci that encode phosphotransferase system (PTS) transporters and phospho-β-glucosidases, which mediate the uptake and deglucosylation of these compounds, respectively. Inactivating these transport and hydrolysis genes abolished or severely reduced growth on PG, establishing the specificity of the loci to distinct groups of PGs. Following intracellular deglucosylation, the aglycones of PGs are externalized, rendering them available for absorption by the host or for further modification by other microbiota taxa. The PG utilization loci are conserved in L. acidophilus and closely related lactobacilli, in correlation with versatile growth on these compounds. Growth on the tested PG appeared more common among human gut lactobacilli than among counterparts from other ecologic niches. The PGs that supported the growth of L. acidophilus were utilized poorly or not at all by other common HGM strains, underscoring the metabolic specialization of L. acidophilus These findings highlight the role of human gut L. acidophilus and select lactobacilli in the bioconversion of glycoconjugated phytochemicals, which is likely to have an important impact on the HGM and human host. IMPORTANCE Thousands of therapeutically active plant-derived compounds are widely present in berries, fruits, nuts, and beverages like tea and wine. The bioactivity and bioavailability of these

  14. Purification of Lactobacillus acidophilus surface-layer protein and its immunomodulatory effects on RAW264.7 cells.

    PubMed

    Zhang, Dandan; Wu, Mengting; Guo, Yuxing; Xun, Mingyue; Wang, Wenwen; Wu, Zhen; Pan, Daodong

    2017-09-01

    Surface-layer proteins (SLP) have been found in the outermost layer of the cell wall in many types of lactobacillus are considered to be an important factor with respect to intestinal immunity. The present study compared the effects of SLP extracted by different concentrations of LiCl and carbamide, and subsequently identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, circular dichroism and differential scanning calorimetry. Furthermore, RAW 264.7 cells were used to evaluate the immunomodulatory effects of SLP. SLP were derived from Lactobacillus acidophilus CICC6074 with a molecular weight of 46 kDa, and consisted of 16.9% α-helix, 42.3% β-sheet, 20.8% β-turns and 22.5% random coils. SLP promoted NO secretion and higher quantities of NO were produced as the SLP concentrations increased. SLP concentrations over 50 µg mL -1 significantly decreased the amount of tumor necrosis factor-α secreted by RAW264.7 cells. SLP can trigger immunomodulatory effects in RAW 264.7 cells. This provides crucial information that will enable the further use of L. acidophilus in food, medicine and other products. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  15. Combined effect of Lactobacillus acidophilus and β-cyclodextrin on serum cholesterol in pigs.

    PubMed

    Alonso, L; Fontecha, J; Cuesta, P

    2016-01-14

    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% β-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.

  16. Impact of oral Lactobacillus acidophilus gavage on rooster seminal and cloacal Lactobacilli concentrations.

    PubMed

    Kiess, A S; Hirai, J H; Triplett, M D; Parker, H M; McDaniel, C D

    2016-08-01

    The use of antibiotics in poultry is being heavily scrutinized, therefore alternatives such as probiotics are being investigated. Lactobacilli spp. are a commonly used bacteria in formulating probiotics, and the addition of Lactobacilli to broiler diets has demonstrated increased growth rates, stimulated immune systems, and reduced pathogen loads in the gastro-intestinal tract ( GI: ) tract. However, previous research has shown that when rooster semen is directly exposed to Lactobacillus acidophilus (L. acidophilus) sperm quality is reduced. Therefore, the objective of the current study was to determine if oral administration of L. acidophilus increases the concentration of Lactobacilli in semen as well as the cloaca. A total of 30 roosters were used: 15 roosters were gavaged with 1X PBS (Control) and 15 roosters were gavaged with 10(7) cfu/mL of L. acidophilus (Treated). All roosters were gavaged for 14 consecutive days. Semen was collected on a 3 d interval, and cloacal swabs were collected on a 2 d interval, beginning on the first day prior to oral administration. Semen and cloacal swabs were serial diluted, and 100 μL of each dilution was then plated on Man, Rogosa, Sharpe ( MRS: ) agar plates. All plates were incubated for 48 h at 37°C under anaerobic conditions and counted. All Lactobacilli counts were first log transformed, then log transformed (day 0) pre-counts were subtracted from the log transformed day counts providing log differences for the analysis. Seminal Lactobacilli counts were not altered by treatments. However, the main effect of treatment (P = 0.026) for cloacal counts indicated that roosters gavaged with Lactobacilli yielded higher counts than the controls. Additionally, cloaca samples also demonstrated a treatment by day interaction trend (P = 0.082), where Lactobacilli was higher in the L. acidophilus gavaged roosters than the controls only on days 3, 5, 13, and 15. In conclusion, the addition of L. acidophilus to the male breeder diet

  17. An Extracellular Cell-Attached Pullulanase Confers Branched α-Glucan Utilization in Human Gut Lactobacillus acidophilus

    PubMed Central

    Møller, Marie S.; Rasmussen, Kasper Bøwig; Cypryk, Wojciech; Celebioglu, Hasan Ufuk; Klaenhammer, Todd R.; Svensson, Birte

    2017-01-01

    ABSTRACT Of the few predicted extracellular glycan-active enzymes, glycoside hydrolase family 13 subfamily 14 (GH13_14) pullulanases are the most common in human gut lactobacilli. These enzymes share a unique modular organization, not observed in other bacteria, featuring a catalytic module, two starch binding modules, a domain of unknown function, and a C-terminal surface layer association protein (SLAP) domain. Here, we explore the specificity of a representative of this group of pullulanases, Lactobacillus acidophilus Pul13_14 (LaPul13_14), and its role in branched α-glucan metabolism in the well-characterized Lactobacillus acidophilus NCFM, which is widely used as a probiotic. Growth experiments with L. acidophilus NCFM on starch-derived branched substrates revealed a preference for α-glucans with short branches of about two to three glucosyl moieties over amylopectin with longer branches. Cell-attached debranching activity was measurable in the presence of α-glucans but was repressed by glucose. The debranching activity is conferred exclusively by LaPul13_14 and is abolished in a mutant strain lacking a functional LaPul13_14 gene. Hydrolysis kinetics of recombinant LaPul13_14 confirmed the preference for short-branched α-glucan oligomers consistent with the growth data. Curiously, this enzyme displayed the highest catalytic efficiency and the lowest Km reported for a pullulanase. Inhibition kinetics revealed mixed inhibition by β-cyclodextrin, suggesting the presence of additional glucan binding sites besides the active site of the enzyme, which may contribute to the unprecedented substrate affinity. The enzyme also displays high thermostability and higher activity in the acidic pH range, reflecting adaptation to the physiologically challenging conditions in the human gut. IMPORTANCE Starch is one of the most abundant glycans in the human diet. Branched α-1,6-glucans in dietary starch and glycogen are nondegradable by human enzymes and constitute a

  18. Improved bioavailability of dietary phenolic acids in whole grain barley and oat groat following fermentation with probiotic Lactobacillus acidophilus , Lactobacillus johnsonii , and Lactobacillus reuteri.

    PubMed

    Hole, Anastasia S; Rud, Ida; Grimmer, Stine; Sigl, Stefanie; Narvhus, Judith; Sahlstrøm, Stefan

    2012-06-27

    The aim of this study was to improve the bioavailability of the dietary phenolic acids in flours from whole grain barley and oat groat following fermentation with lactic acid bacteria (LAB) exhibiting high feruloyl esterase activity (FAE). The highest increase of free phenolic acids was observed after fermentation with three probiotic strains, Lactobacillus johnsonii LA1, Lactobacillus reuteri SD2112, and Lactobacillus acidophilus LA-5, with maximum increases from 2.55 to 69.91 μg g(-1) DM and from 4.13 to 109.42 μg g(-1) DM in whole grain barley and oat groat, respectively. Interestingly, higher amounts of bound phenolic acids were detected after both water treatment and LAB fermentation in whole grain barley, indicating higher bioaccessibility, whereas some decrease was detected in oat groat. To conclude, cereal fermentation with specific probiotic strains can lead to significant increase of free phenolic acids, thereby improving their bioavailability.

  19. Effects of cadmium on the growth and uptake of cadmium by microorganisms. [Esherichia coli; Bacillus cereus; Lactobacillus acidophilus; Staphylococcus aureus; Streptococcus faecalis; Actinomyces niger

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Doyle, J.J.; Marshall, R.T.; Pfander, W.H.

    1975-01-01

    Six species of microorganisms, Escherichia coli, Bacillus cereus, Lactobacillus acidophilus, Staphylococcus aureus, Streptococcus faecalis and Actinomyces niger, were grown under suitable conditions in appropriate media. Cadmium chloride was added to provide 0, 5, 10, 20, 40, and 80 ..mu..g of Cd per ml. At 40 and 80 ..mu..g of Cd per ml, E. coli and B. cereus grew well and the other species were repressed. Cd uptake patterns differed significantly among the species tested. The significance of these data with respect to Cd in food chains is discussed. 14 references, 3 tables.

  20. Effects of Lactobacillus acidophilus on gut microbiota composition in broilers challenged with Clostridium perfringens

    PubMed Central

    Wang, Weiwei; Liu, Dan; Guo, Yuming

    2017-01-01

    This study shows the effects of dietary supplementation with Lactobacillus acidophilus on the gut microbiota of broiler chickens challenged with Clostridium perfringens infection during a 21-day period according to pyrosequencing of the 16S ribosomal RNA gene. In a 2 × 2 factorial arrangement of treatments, 308 1-day-old male Arbor Acres broiler chicks were analyzed for the effects of the probiotic (groups without or with L. acidophilus supplementation), pathogen challenge (groups without or with C. perfringens), and the effects of interaction. The infection decreased the number of Observed species, Chao1, and ACE of ileal microbiota and increased Chao1 of cecal microbiota of broilers, whereas L. acidophilus supplementation decreased the Shannon index of the ileal microbiota. Shannon index and Simpson indices were lower in the ileal microbiota than in the cecal microbiota. In the ileal microbiota, the control group had higher relative abundance of Lachnospiraceae and Ruminococcaceae in comparison with the other groups; however, the relative abundance of Gammaproteobacteria was significantly higher in the challenge group than in the other groups. C. perfringens infection tended to increase lactate concentration and decreasedconcentrations of formate, acetate and propionate in the ileum; decreased isobutyrate concentration; and tended to decrease isovalerate concentration in the cecum. Besides, L. acidophilus supplementation increased the concentration of lactate and butyrate and decreased concentrations of formate and propionate in the ileum, and increased concentrations of lactate and valerate in the cecum. In conclusion, C. perfringens infection and/or dietary supplementation with L. acidophilus modulated the relative abundance of some bacteria taxa, and the L. acidophilus supplementation helped to restore the microbial community disrupted by C. perfringens infection. PMID:29190649

  1. Effects of Lactobacillus acidophilus on gut microbiota composition in broilers challenged with Clostridium perfringens.

    PubMed

    Li, Zhui; Wang, Weiwei; Liu, Dan; Guo, Yuming

    2017-01-01

    This study shows the effects of dietary supplementation with Lactobacillus acidophilus on the gut microbiota of broiler chickens challenged with Clostridium perfringens infection during a 21-day period according to pyrosequencing of the 16S ribosomal RNA gene. In a 2 × 2 factorial arrangement of treatments, 308 1-day-old male Arbor Acres broiler chicks were analyzed for the effects of the probiotic (groups without or with L. acidophilus supplementation), pathogen challenge (groups without or with C. perfringens), and the effects of interaction. The infection decreased the number of Observed species, Chao1, and ACE of ileal microbiota and increased Chao1 of cecal microbiota of broilers, whereas L. acidophilus supplementation decreased the Shannon index of the ileal microbiota. Shannon index and Simpson indices were lower in the ileal microbiota than in the cecal microbiota. In the ileal microbiota, the control group had higher relative abundance of Lachnospiraceae and Ruminococcaceae in comparison with the other groups; however, the relative abundance of Gammaproteobacteria was significantly higher in the challenge group than in the other groups. C. perfringens infection tended to increase lactate concentration and decreasedconcentrations of formate, acetate and propionate in the ileum; decreased isobutyrate concentration; and tended to decrease isovalerate concentration in the cecum. Besides, L. acidophilus supplementation increased the concentration of lactate and butyrate and decreased concentrations of formate and propionate in the ileum, and increased concentrations of lactate and valerate in the cecum. In conclusion, C. perfringens infection and/or dietary supplementation with L. acidophilus modulated the relative abundance of some bacteria taxa, and the L. acidophilus supplementation helped to restore the microbial community disrupted by C. perfringens infection.

  2. Lactobacillus acidophilus K301 Inhibits Atherogenesis via Induction of 24 (S), 25-Epoxycholesterol-Mediated ABCA1 and ABCG1 Production and Cholesterol Efflux in Macrophages

    PubMed Central

    Kim, Hye Sun; Park, Woo Jung; Kim, Joo-Yun; Chung, Dae Kyun

    2016-01-01

    Lactobacillus acidophilus species are well-known probiotics with the beneficial activity of regulating cholesterol levels. In this study, we showed that L. acidophilus K301 reduced the level of cholesterol through reverse transport in macrophages. L. acidophilus K301 upregulated the mRNA and protein levels of genes such as ATP-binding cassette A1 (ABCA1) and ATP-binding cassette G1 (ABCG1) under the control of liver X receptor (LXR), resulting in increased apoA-I-dependent cholesterol efflux in phorbol 12-myristate 13-acetate (PMA)-differentiated THP-1 cells. L. acidophilus K301 induced both ABCA1 and ABCG1 through the endogenous LXR agonist 24(S), 25-epoxcycholesterol, which is synthesized by intracellular cholesterol synthetic pathways. In vivo studies using L. acidophilus K301-treated ApoE-/- mice showed reduced accumulation of lipoproteins in the arterial lumen. The inhibitory effects of L. acidophilus K301 on accumulation of lipoprotein in atherosclerotic plaques were mediated by the induction of squalene reductase (SQLE) and oxidosqualene cyclase (OSC) and resulted in ABCA1-mediated cholesterol efflux. Taken together, our findings revealed that Lactobacillus acidophilus K301 regulates the expression of genes related to cholesterol reverse transport via the induction of endogenous LXR agonist, suggesting the therapeutic potential of Lactobacillus acidophilus K301 as an anti-atherosclerotic agent. PMID:27120199

  3. Lactobacillus acidophilus K301 Inhibits Atherogenesis via Induction of 24 (S), 25-Epoxycholesterol-Mediated ABCA1 and ABCG1 Production and Cholesterol Efflux in Macrophages.

    PubMed

    Hong, Yi-Fan; Kim, Hangeun; Kim, Hye Sun; Park, Woo Jung; Kim, Joo-Yun; Chung, Dae Kyun

    2016-01-01

    Lactobacillus acidophilus species are well-known probiotics with the beneficial activity of regulating cholesterol levels. In this study, we showed that L. acidophilus K301 reduced the level of cholesterol through reverse transport in macrophages. L. acidophilus K301 upregulated the mRNA and protein levels of genes such as ATP-binding cassette A1 (ABCA1) and ATP-binding cassette G1 (ABCG1) under the control of liver X receptor (LXR), resulting in increased apoA-I-dependent cholesterol efflux in phorbol 12-myristate 13-acetate (PMA)-differentiated THP-1 cells. L. acidophilus K301 induced both ABCA1 and ABCG1 through the endogenous LXR agonist 24(S), 25-epoxcycholesterol, which is synthesized by intracellular cholesterol synthetic pathways. In vivo studies using L. acidophilus K301-treated ApoE-/- mice showed reduced accumulation of lipoproteins in the arterial lumen. The inhibitory effects of L. acidophilus K301 on accumulation of lipoprotein in atherosclerotic plaques were mediated by the induction of squalene reductase (SQLE) and oxidosqualene cyclase (OSC) and resulted in ABCA1-mediated cholesterol efflux. Taken together, our findings revealed that Lactobacillus acidophilus K301 regulates the expression of genes related to cholesterol reverse transport via the induction of endogenous LXR agonist, suggesting the therapeutic potential of Lactobacillus acidophilus K301 as an anti-atherosclerotic agent.

  4. Production of 13S-hydroxy-9(Z)-octadecenoic acid from linoleic acid by whole recombinant cells expressing linoleate 13-hydratase from Lactobacillus acidophilus.

    PubMed

    Park, Ji-Young; Lee, Seon-Hwa; Kim, Kyoung-Rok; Park, Jin-Byung; Oh, Deok-Kun

    2015-08-20

    Linoleate 13-hydratase from Lactobacillus acidophilus LMG 11470 converted linoleic acid to hydroxyl fatty acid, which was identified as 13S-hydroxy-9(Z)-octadecenoic acid (13-HOD) by GC-MS and NMR. The expression of linoleate 13-hydratase gene in Escherichia coli was maximized by using pACYC plasmid and super optimal broth with catabolite repression (SOC) medium containing 40mM Mg(2+). To optimize induction conditions, recombinant cells were cultivated at 37°C, 1mM isopropyl-β-d-thiogalactopyranoside was added at 2h, and the culture was further incubated at 16°C for 18h. Recombinant cells expressing linoleate 13-hydratase from L. acidophilus were obtained under the optimized expression conditions and used for 13-HOD production from linoleic acid. The optimal reaction conditions were pH 6.0, 40°C, 0.25% (v/v) Tween 40, 25gl(-1) cells, and 100gl(-1) linoleic acid, and under these conditions, whole recombinant cells produced 79gl(-1) 13-HOD for 3h with a conversion yield of 79% (w/w), a volumetric productivity of 26.3gl(-1)h(-1), and a specific productivity of 1.05g g-cells(-1)h(-1). To the best of our knowledge, the recombinant cells produced hydroxy fatty acid with the highest concentration and productivity reported so far. Crown Copyright © 2015. Published by Elsevier B.V. All rights reserved.

  5. Microencapsulation of Lactobacillus acidophilus NCFM using polymerized whey proteins as wall material.

    PubMed

    Jiang, Yujun; Zheng, Zhe; Zhang, Tiehua; Hendricks, Gregory; Guo, Mingruo

    2016-09-01

    Survivability of probiotics in foods is essential for developing functional food containing probiotics. We investigated polymerized whey protein (PWP)-based microencapsulation process which is developed for protecting probiotics like Lactobacillus acidophilus NCFM and compared with the method using sodium alginate (SA). The entrapment rate was 89.3 ± 4.8% using PWP, while it was 73.2 ± 1.4% for SA. The microencapsulated NCFM by PWP and SA were separately subjected to digestion juices and post-fermentation storage of fermented cows' and goats' milk using the encapsulated culture. The log viable count of NCFM in PWP-based microencapsulation was 4.56, compared with that of 4.26 in SA-based ones and 3.13 for free culture. Compared with using SA as wall material, PWP was more effective in protecting probiotic. Microencapsulation of L. acidophilus NCFM using PWP as wall material can be exploited in the development of fermented dairy products with better survivability of probiotic organism.

  6. Lactobacillus acidophilus Metabolizes Dietary Plant Glucosides and Externalizes Their Bioactive Phytochemicals

    PubMed Central

    Theilmann, Mia C.; Nielsen, Kristian Fog; Klaenhammer, Todd R.

    2017-01-01

    ABSTRACT Therapeutically active glycosylated phytochemicals are ubiquitous in the human diet. The human gut microbiota (HGM) modulates the bioactivities of these compounds, which consequently affect host physiology and microbiota composition. Despite a significant impact on human health, the key players and the underpinning mechanisms of this interplay remain uncharacterized. Here, we demonstrate the growth of Lactobacillus acidophilus on mono- and diglucosyl dietary plant glycosides (PGs) possessing small aromatic aglycones. Transcriptional analysis revealed the upregulation of host interaction genes and identified two loci that encode phosphotransferase system (PTS) transporters and phospho-β-glucosidases, which mediate the uptake and deglucosylation of these compounds, respectively. Inactivating these transport and hydrolysis genes abolished or severely reduced growth on PG, establishing the specificity of the loci to distinct groups of PGs. Following intracellular deglucosylation, the aglycones of PGs are externalized, rendering them available for absorption by the host or for further modification by other microbiota taxa. The PG utilization loci are conserved in L. acidophilus and closely related lactobacilli, in correlation with versatile growth on these compounds. Growth on the tested PG appeared more common among human gut lactobacilli than among counterparts from other ecologic niches. The PGs that supported the growth of L. acidophilus were utilized poorly or not at all by other common HGM strains, underscoring the metabolic specialization of L. acidophilus. These findings highlight the role of human gut L. acidophilus and select lactobacilli in the bioconversion of glycoconjugated phytochemicals, which is likely to have an important impact on the HGM and human host. PMID:29162708

  7. Antibacterial activity of Lactobacillus acidophilus strains isolated from honey marketed in Malaysia against selected multiple antibiotic resistant (MAR) Gram-positive bacteria.

    PubMed

    Aween, Mohamed Mustafa; Hassan, Zaiton; Muhialdin, Belal J; Eljamel, Yossra A; Al-Mabrok, Asma Saleh W; Lani, Mohd Nizam

    2012-07-01

    A total of 32 lactic acid bacteria (LAB) were isolated from 13 honey samples commercially marketed in Malaysia, 6 strains identified as Lactobacillus acidophilus by API CHL50. The isolates had antibacterial activities against multiple antibiotic resistant's Staphylococcus aureus (25 to 32 mm), Staphylococcus epidermis (14 to 22 mm) and Bacillus subtilis (12 to 19 mm) in the agar overlay method after 24 h incubation at 30 °C. The crude supernatant was heat stable at 90 °C and 121 °C for 1 h. Treatment with proteinase K and RNase II maintained the antimicrobial activity of all the supernatants except sample H006-A and H010-G. All the supernatants showed antimicrobial activities against target bacteria at pH 3 and pH 5 but not at pH 6 within 72 h incubation at 30 °C. S. aureus was not inhibited by sample H006-A isolated from Libyan honey and sample H008-D isolated from Malaysian honey at pH 5, compared to supernatants from other L. acidophilus isolates. The presence of different strains of L. acidophilus in honey obtained from different sources may contribute to the differences in the antimicrobial properties of honey. © 2012 Institute of Food Technologists®

  8. Biochemical characterization of a recombinant Lactobacillus acidophilus strain expressing exogenous FomA protein.

    PubMed

    Ma, Li; Li, Fei; Zhang, Xiangyu; Feng, Xiping

    2018-04-30

    In previous research, to combine the immunogenicity of Fusobacterium nucleatum (F. nucleatum) and the probiotic properties of Lactobacillus acidophilus (L. acidophilus), we constructed a FomA-expressing L. acidophilus strain and assessed its immunogenicity. Our findings indicated that oral administration of the recombinant L. acidophilus strain reduced the risk of periodontal infection by Porphyromonas gingivalis (P. gingivalis) and F. nucleatum. However, because the exogenous FomA is an heterologous protein for the original bacterium, in this study, we assessed whether the biochemical characteristics of the recombinant L. acidophilus strain change due to the expression of the exogenous FomA protein. To test the biochemical characteristics of a recombinant L. acidophilus strain expressing exogenous FomA and assess its antibiotic sensitivity. We assessed the colony morphology, growth, acid production, and carbohydrate fermentation abilities of the recombinant L. acidophilus strain. In addition, we tested the adhesive ability and antimicrobial activity of the recombinant and assessed its antibiotic sensitivity through a drug susceptibility test. The experimental results showed that the colony and microscopic morphology of the recombinant L. acidophilus strain was consistent with the original strain, and the recombinant strain grew well when cultured under aerobic or anaerobic conditions, exhibiting a growth rate that was identical to that of the standard strain. Similarly, the supernatants of the recombinant L. acidophilus can inhibit the growth of E. coli and P. gingivalis at different concentrations, and the recombinant strain displayed essentially the same drug sensitivity profile as the original L. acidophilus. However, to our surprise, the recombinant strains exhibited a greater adhesion ability than the reference strain. Our study demonstrated that, in addition to an increased adhesion ability, the recombinant L. acidophilus strain maintained the basic

  9. Effect of dietary supplementation with Lactobacillus acidophilus D2/CSL (CECT 4529) on caecum microbioma and productive performance in broiler chickens.

    PubMed

    De Cesare, Alessandra; Sirri, Federico; Manfreda, Gerardo; Moniaci, Paola; Giardini, Alberto; Zampiga, Marco; Meluzzi, Adele

    2017-01-01

    This study examines the effects of the dietary supplementation with Lactobacillus acidophilus D2/CSL (CECT 4529) (LA) on productive performances, incidence of foot pad dermatitis and caecum microbioma in broiler chickens. A total of 1,100 one-day old male Ross 308 chicks were divided into 2 groups of 16 replicates with 25 birds each and reared from 1-41 d. One group was fed a basal diet (CON) and the other group the same diet supplemented with LA. Caecum contents were collected from 4 selected birds at day one and 5 selected birds at the end of the rearing period. Then, they were submitted to DNA extraction and whole DNA shotgun metagenomic sequencing. Overall, the LA supplementation produced a significant beneficial effect on body weight gain between 15-28 d and improved feed conversion rate in the overall period. On the contrary, litter moisture, pH and incidence of the foot pad lesions were not affected by LA. Birds treated with LA showed a lower occurrence of pasty vent at both 14 and 28 d. At the end of the rearing period, Lachanospiraceae were significantly higher in LA birds in comparison to CON (17.07 vs 14.39%; P = 0.036). Moreover, Ruminococcus obeum, Clostridium clostridioforme, Roseburia intestinalis, Lachnospiraceae bacterium 14-2T and Coprococcus eutactus were significantly higher in LA birds in comparison to CON. The relative abundance of Lactobacillus acidophilus was comparable between LA and CON groups. However, a positive effect was observed in relation to the metabolic functions in the treated group, with particular reference to the higher abundance of β-glucosidase. In conclusion, the LA supplementation improved broiler productive performances and metabolic functions promoting animal health.

  10. Effect of dietary supplementation with Lactobacillus acidophilus D2/CSL (CECT 4529) on caecum microbioma and productive performance in broiler chickens

    PubMed Central

    De Cesare, Alessandra; Manfreda, Gerardo; Moniaci, Paola; Giardini, Alberto; Zampiga, Marco; Meluzzi, Adele

    2017-01-01

    This study examines the effects of the dietary supplementation with Lactobacillus acidophilus D2/CSL (CECT 4529) (LA) on productive performances, incidence of foot pad dermatitis and caecum microbioma in broiler chickens. A total of 1,100 one-day old male Ross 308 chicks were divided into 2 groups of 16 replicates with 25 birds each and reared from 1–41 d. One group was fed a basal diet (CON) and the other group the same diet supplemented with LA. Caecum contents were collected from 4 selected birds at day one and 5 selected birds at the end of the rearing period. Then, they were submitted to DNA extraction and whole DNA shotgun metagenomic sequencing. Overall, the LA supplementation produced a significant beneficial effect on body weight gain between 15–28 d and improved feed conversion rate in the overall period. On the contrary, litter moisture, pH and incidence of the foot pad lesions were not affected by LA. Birds treated with LA showed a lower occurrence of pasty vent at both 14 and 28 d. At the end of the rearing period, Lachanospiraceae were significantly higher in LA birds in comparison to CON (17.07 vs 14.39%; P = 0.036). Moreover, Ruminococcus obeum, Clostridium clostridioforme, Roseburia intestinalis, Lachnospiraceae bacterium 14-2T and Coprococcus eutactus were significantly higher in LA birds in comparison to CON. The relative abundance of Lactobacillus acidophilus was comparable between LA and CON groups. However, a positive effect was observed in relation to the metabolic functions in the treated group, with particular reference to the higher abundance of β-glucosidase. In conclusion, the LA supplementation improved broiler productive performances and metabolic functions promoting animal health. PMID:28472118

  11. Efficacy profiles for different concentrations of Lactobacillus acidophilus in experimental colitis.

    PubMed

    Chen, Lin-Lin; Zou, Yi-You; Lu, Fang-Gen; Li, Fu-Jun; Lian, Guang-Hui

    2013-08-28

    To determine the efficacy profiles of different concentrations of Lactobacillus acidophilus (L. acidophilus) for treating colitis using an experimental murine model. Colitis was established in 64 BALB/c mice by adding 5% dextran sodium sulfate (DSS) to the drinking water and allowing ad libitum access for 7 d. The mice were then randomly divided into the following control and experimental model groups (n = 8 each; day 0): untreated model control; negative-treatment model control (administered gavage of 1 mL/10 g normal saline); experimental-treatment models C4-C8 (administered gavage of 10(4), 10(5), 10(6), 10(7), or 10(8) CFU/10 g L. acidophilus, respectively); positive-treatment model control (administration of the anti-inflammatory agent prednisone acetate at 45 μg/10 g). Eight mice given regular water (no DSS) and no subsequent treatments served as the normal control group. Body weight, fecal traits, and presence of fecal occult blood were assessed daily. All animals were sacrificed on post-treatment day 7 to measure colonic length, perform histological scoring, and quantify the major bacteria in the proximal and distal colon. Intergroup differences were determined by one-way ANOVA and post-hoc Student-Newman-Keuls comparison. All treatments (L. acidophilus and prednisone acetate) protected against colitis-induced weight loss (P < 0.05 vs model and normal control groups). The extent of colitis-induced colonic shortening was significantly reduced by all treatments (prednisone acetate > C4 > C5 > C7 > C8 > C6; P < 0.05 vs untreated model group), and the C6 group showed colonic length similar to that of the normal control group (P > 0.05). The C6 group also had the lowest disease activity index scores among the model groups. The bacterial profiles in the proximal colon were similar between all of the experimental-treatment model groups (all P > 0.05). In contrast, the bacterial profile in the distal colon of the C6 group showed the distinctive features (P < 0

  12. Oral Immunization with Recombinant Lactobacillus acidophilus Expressing the Adhesin Hp0410 of Helicobacter pylori Induces Mucosal and Systemic Immune Responses

    PubMed Central

    Hongying, Fan; Xianbo, Wu; Fang, Yu; Yang, Bai

    2014-01-01

    Helicobacter pylori infection is relatively common worldwide and is closely related to gastric mucosa-associated lymphoid tissue (MALT) lymphoma, chronic gastritis, and stomach ulcers. Therefore, a safe and effective method for preventing H. pylori infection is urgently needed. Given that developing an effective vaccine against H. pylori is one of the best alternatives, H. pylori adhesin Hp0410 was expressed in the food-grade bacterium Lactobacillus acidophilus. The recombinant live bacterial vaccine was then used to orally vaccinate mice, and the immunoprotective effects of Hp0410-producing strains were investigated. H. pylori colonization in the stomach of mice immunized with the recombinant L. acidophilus was significantly reduced, in comparison with that in control groups. Furthermore, mucosal secretory IgA antibodies were elicited in the mucosal tissue of mice immunized with the recombinant bacteria, and specific anti-Hp0410 IgG responses were also detected in mouse serum. There was a significant increase in the level of protection against gastric Helicobacter infection following a challenge with H. pylori Sydney strain 1 (SS1). Our results collectively indicate that adhesin Hp0410 is a promising candidate vaccine antigen, and recombinant L. acidophilus expressing Hp0410 is likely to constitute an effective, low-cost, live bacterial vaccine against H. pylori. PMID:24285819

  13. An Extracellular Cell-Attached Pullulanase Confers Branched α-Glucan Utilization in Human Gut Lactobacillus acidophilus.

    PubMed

    Møller, Marie S; Goh, Yong Jun; Rasmussen, Kasper Bøwig; Cypryk, Wojciech; Celebioglu, Hasan Ufuk; Klaenhammer, Todd R; Svensson, Birte; Abou Hachem, Maher

    2017-06-15

    Of the few predicted extracellular glycan-active enzymes, glycoside hydrolase family 13 subfamily 14 (GH13_14) pullulanases are the most common in human gut lactobacilli. These enzymes share a unique modular organization, not observed in other bacteria, featuring a catalytic module, two starch binding modules, a domain of unknown function, and a C-terminal surface layer association protein (SLAP) domain. Here, we explore the specificity of a representative of this group of pullulanases, Lactobacillus acidophilus Pul13_14 ( La Pul13_14), and its role in branched α-glucan metabolism in the well-characterized Lactobacillus acidophilus NCFM, which is widely used as a probiotic. Growth experiments with L. acidophilus NCFM on starch-derived branched substrates revealed a preference for α-glucans with short branches of about two to three glucosyl moieties over amylopectin with longer branches. Cell-attached debranching activity was measurable in the presence of α-glucans but was repressed by glucose. The debranching activity is conferred exclusively by La Pul13_14 and is abolished in a mutant strain lacking a functional La Pul13_14 gene. Hydrolysis kinetics of recombinant La Pul13_14 confirmed the preference for short-branched α-glucan oligomers consistent with the growth data. Curiously, this enzyme displayed the highest catalytic efficiency and the lowest K m reported for a pullulanase. Inhibition kinetics revealed mixed inhibition by β-cyclodextrin, suggesting the presence of additional glucan binding sites besides the active site of the enzyme, which may contribute to the unprecedented substrate affinity. The enzyme also displays high thermostability and higher activity in the acidic pH range, reflecting adaptation to the physiologically challenging conditions in the human gut. IMPORTANCE Starch is one of the most abundant glycans in the human diet. Branched α-1,6-glucans in dietary starch and glycogen are nondegradable by human enzymes and constitute a

  14. Modulation of the Lactobacillus acidophilus La-5 lipidome by different growth conditions.

    PubMed

    Hansen, Marie-Louise R W; Clausen, Anders; Ejsing, Christer S; Risbo, Jens

    2015-10-01

    Probiotics are bacteria used in the food industry due to their potential health benefits. In this study, the plasma membrane of the probiotic Lactobacillus acidophilus La-5 was investigated using state-of-the-art high-resolution shotgun lipidomics. Comparisons of the lipidome of the plasma membrane were done after altering the fatty acid composition by supplementing L. acidophilus La-5 with saturated, mono-, di- and tri-unsaturated fatty acids during fermentation. The plasma membrane with the highest degree of saturation resulted in a lipid composition with the highest proportion of cardiolipin (CL) and lowest proportion of monolysocardiolipin (MLCL). No significant changes were found for other lipid classes. The bacteria grown with di- and tri-unsaturated fatty acids were expected to have more unsaturated plasma membranes than bacteria grown with mono-unsaturated fatty acids. This was also the case for MLCL, but the numbers of double bonds for CL were quite similar for these three samples. The results indicate that L. acidophilus La-5 possesses a molecular mechanism for remodelling and optimizing the fatty acid composition of CL and MLCL species and the molar ratio of CL and MLCL. This study contributes new knowledge on the previously uninvestigated lipidome of L. acidophilus La-5.

  15. The effect of a probiotic strain (Lactobacillus acidophilus) on the plaque formation of oral Streptococci.

    PubMed

    Tahmourespour, Arezoo; Kermanshahi, Rooha Kasra

    2011-02-01

    The objective of this study was to investigate the ability of biofilm formation among mutans and non mutans oral streptococci and to determine the effect of Lactobacillus acidophilus DSM 20079 as a probiotic strain on the adhesion of selected streptococcal strains on the surfaces. The sample comprised 40 isolates of oral streptococci from dental plaque and caries of volunteer persons. Streptococcus mutans ATCC35668 (no24) was as an standard strain. The probiotic strain was Lactobacillus acidophilus DSM 20079. The ability of biofilm formation was investigated with colorimetric method and the strongest isolates were selected. Then the effect of probiotic strain on the adhesion of streptococci isolates was determined in polystyrene microtiter plate simultaneously and 30 minutes before streptococci entrance to the system. The results showed that 42% of mutans streptococci were strongly adherent (SA) and in non mutans streptococci, only 23.5% of isolates were found strongly adherent. The strong biofilm forming bacterium isolated was Streptococcus mutans strain22. In the next step, in the presence of probiotic strain the streptococcal adhesion were reduced, and this reduction was non significantly stronger if the probiotic strain was inoculated to the system before the oral bacteria. The Lactobacillus acidophilus had more effect on adherence of mutans streptococci than non mutans streptococci with significant difference (p < 0.05). Adhesion reduction is likely due to bacterial interactions and colonization of adhesion sites with probiotic strain before the presence of streptococci. Adhesion reduction can be an effective way on decreasing cariogenic potential of oral streptococci.

  16. Inhibition of initial adhesion of uropathogenic Enterococcus faecalis by biosurfactants from Lactobacillus isolates.

    PubMed

    Velraeds, M M; van der Mei, H C; Reid, G; Busscher, H J

    1996-06-01

    In this study, 15 Lactobacillus isolates were found to produce biosurfactants in the mid-exponential and stationary growth phases. The stationary-phase biosurfactants from lactobacillus casei subsp. rhamnosus 36 and ATCC 7469, Lactobacillus fermentum B54, and Lactobacillus acidophilus RC14 were investigated further to determine their capacity to inhibit the initial adhesion of uropathogenic Enterococcus faecalis 1131 to glass in a parallel-plate flow chamber. The initial deposition rate of E. faecalis to glass with an adsorbed biosurfactant layer from L. acidophilus RC14 or L. fermentum B54 was significantly decreased by approximately 70%, while the number of adhering enterococci after 4 h of adhesion was reduced by an average of 77%. The surface activity of the biosurfactants and their activity inhibiting the initial adhesion of E. faecalis 1131 were retained after dialysis (molecular weight cutoff, 6,000 to 8,000) and freeze-drying. Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy revealed that the freeze-dried biosurfactants from L. acidophilus RC14 and L. fermentum B54 were richest in protein, while those from L. casei subsp. rhamnosus 36 and ATCC 7469 had relatively high polysaccharide and phosphate contents.

  17. Inhibition of initial adhesion of uropathogenic Enterococcus faecalis by biosurfactants from Lactobacillus isolates.

    PubMed Central

    Velraeds, M M; van der Mei, H C; Reid, G; Busscher, H J

    1996-01-01

    In this study, 15 Lactobacillus isolates were found to produce biosurfactants in the mid-exponential and stationary growth phases. The stationary-phase biosurfactants from lactobacillus casei subsp. rhamnosus 36 and ATCC 7469, Lactobacillus fermentum B54, and Lactobacillus acidophilus RC14 were investigated further to determine their capacity to inhibit the initial adhesion of uropathogenic Enterococcus faecalis 1131 to glass in a parallel-plate flow chamber. The initial deposition rate of E. faecalis to glass with an adsorbed biosurfactant layer from L. acidophilus RC14 or L. fermentum B54 was significantly decreased by approximately 70%, while the number of adhering enterococci after 4 h of adhesion was reduced by an average of 77%. The surface activity of the biosurfactants and their activity inhibiting the initial adhesion of E. faecalis 1131 were retained after dialysis (molecular weight cutoff, 6,000 to 8,000) and freeze-drying. Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy revealed that the freeze-dried biosurfactants from L. acidophilus RC14 and L. fermentum B54 were richest in protein, while those from L. casei subsp. rhamnosus 36 and ATCC 7469 had relatively high polysaccharide and phosphate contents. PMID:8787394

  18. Lactobacillus rhamnosus HN001 and Lactobacillus acidophilus La-14 Attenuate Gardnerella vaginalis-Infected Bacterial Vaginosis in Mice.

    PubMed

    Jang, Se-Eun; Jeong, Jin-Ju; Choi, Su-Young; Kim, Hyunji; Han, Myung Joo; Kim, Dong-Hyun

    2017-05-23

    Oral administration of a probiotic mixture (PM; Respecta ® ) consisting of Lactobacillus rhamnosus HN001 (L1), Lactobacillus acidophilus La-14 (L2), and lactoferrin RCXTM results in colonization of these probiotics in the vagina of healthy women. Therefore, we examined whether vaginal colonization of the PM ingredients L1 and L2 could attenuate bacterial vaginosis (BV). BV was induced in mice via β-estradiol-3-benzoate-induced immunosuppression and intravaginal inoculation with Gardnerella vaginalis (GV). Inflammatory markers were analyzed using enzyme-linked immunosorbent assay, immunoblotting, quantitative polymerase chain reaction, and flow cytometry. Oral or intravaginal administration of PM resulted in colonization of L1 and L2 in the vagina. Oral or intravaginal administration of L1, L2, or PM significantly inhibited GV-induced epithelial cell disruption, myeloperoxidase activity, NF-κB activation, and IL-1β and TNF-α expression ( p < 0.05). Administration of these probiotics also inhibited IL-17 and RORγt expression but increased IL-10 and Foxp3 expression. Of these probiotics, L2 most effectively attenuated GV-induced BV, followed by L1 and PM. Oral administration was more effective against GV-induced BV than intravaginal administration. L1 and L2 also significantly inhibited the adherence of GV to HeLa cells (a human cervical cancer cell line) and GV growth in vitro. In addition, L1 and L2 inhibited lipopolysaccharide-induced NF-κB activation in macrophages and the differentiation of splenocytes into Th17 cells in vitro, but increased their differentiation into Treg cells. Our study suggests that L1, L2, and PM attenuated GV-induced vaginosis by regulating both vaginal and systemic innate and adaptive immune responses rather than direct competition or killing of GV in the vagina.

  19. Lactobacillus rhamnosus HN001 and Lactobacillus acidophilus La-14 Attenuate Gardnerella vaginalis-Infected Bacterial Vaginosis in Mice

    PubMed Central

    Jang, Se-Eun; Jeong, Jin-Ju; Choi, Su-Young; Kim, Hyunji; Han, Myung Joo; Kim, Dong-Hyun

    2017-01-01

    Oral administration of a probiotic mixture (PM; Respecta®) consisting of Lactobacillus rhamnosus HN001 (L1), Lactobacillus acidophilus La-14 (L2), and lactoferrin RCXTM results in colonization of these probiotics in the vagina of healthy women. Therefore, we examined whether vaginal colonization of the PM ingredients L1 and L2 could attenuate bacterial vaginosis (BV). BV was induced in mice via β-estradiol-3-benzoate-induced immunosuppression and intravaginal inoculation with Gardnerella vaginalis (GV). Inflammatory markers were analyzed using enzyme-linked immunosorbent assay, immunoblotting, quantitative polymerase chain reaction, and flow cytometry. Oral or intravaginal administration of PM resulted in colonization of L1 and L2 in the vagina. Oral or intravaginal administration of L1, L2, or PM significantly inhibited GV-induced epithelial cell disruption, myeloperoxidase activity, NF-κB activation, and IL-1β and TNF-α expression (p < 0.05). Administration of these probiotics also inhibited IL-17 and RORγt expression but increased IL-10 and Foxp3 expression. Of these probiotics, L2 most effectively attenuated GV-induced BV, followed by L1 and PM. Oral administration was more effective against GV-induced BV than intravaginal administration. L1 and L2 also significantly inhibited the adherence of GV to HeLa cells (a human cervical cancer cell line) and GV growth in vitro. In addition, L1 and L2 inhibited lipopolysaccharide-induced NF-κB activation in macrophages and the differentiation of splenocytes into Th17 cells in vitro, but increased their differentiation into Treg cells. Our study suggests that L1, L2, and PM attenuated GV-induced vaginosis by regulating both vaginal and systemic innate and adaptive immune responses rather than direct competition or killing of GV in the vagina. PMID:28545241

  20. Strain-specific detection of orally administered canine jejunum-dominated Lactobacillus acidophilus LAB20 in dog faeces by real-time PCR targeted to the novel surface layer protein.

    PubMed

    Tang, Y; Saris, P E J

    2013-10-01

    Lactobacillus acidophilus LAB20 has potential to be a probiotic strain because it can be present at high numbers in the jejunum of dog. To specifically detect LAB20 from dog faecal samples, a real-time PCR protocol was developed targeting the novel surface (S) layer protein gene of LAB20. The presence of S-layer protein was verified by N-terminal sequencing of the approximately 50-kDa major band from SDS-PAGE gel. The corresponding S-layer gene was amplified by inverse PCR using homology to known S-layers and sequenced. This novel S-layer protein has low sequence similarity to other S-layer proteins in the N-terminal region (32-211 aa, 7-39%). This enabled designing strain-specific PCR primers. The primer set was utilized to study intestinal persistence of LAB20 in dog that was fed with LAB20 fermented milk for 5 days. The results showed that LAB20 can be detected from dog faecal sample after 6 weeks with 10(4·53)  DNA copies g(-1) postadministration. It suggested that LAB20 could be a good candidate to study the mechanism behind its persistence and dominance in dog intestine and maybe utilize it as a probiotic for canine. A real-time PCR method was developed to detect Lactobacillus acidophilus LAB20, a strain that was previously found dominant in canine gastrointestinal (GI) tract. The quantitative detection was based on targeting to variation region of a novel S-layer protein found in LAB20, allowing to specifically enumerate LAB20 from dog faeces. The results showed that the real-time PCR method was sensitive enough to be used in later intervention studies. Interestingly, LAB20 was found to persist in dog GI tract for 6 weeks. Therefore, LAB20 could be a good candidate to study its colonization and potentially utilize as a canine probiotic. © 2013 The Society for Applied Microbiology.

  1. The protective effect of recombinant FomA-expressing Lactobacillus acidophilus against periodontal infection.

    PubMed

    Ma, Li; Ding, Qinfeng; Feng, Xiping; Li, Fei

    2013-10-01

    A number of studies have shown that the outer membrane protein FomA found in Fusobacterium nucleatum demonstrates great potential as an immune target for combating periodontitis. Lactobacillus acidophilus is a useful antigen delivery vehicle for mucosal immunisation, and previous studies by our group have shown that L. acidophilus acts as a protective factor in periodontal health. In this study, making use of the immunogenicity of FomA and the probiotic properties of L. acidophilus, we constructed a recombinant form of L. acidophilus expressing the FomA protein and detected the FomA-specific IgG in the serum and sIgA in the saliva of mice through oral administration with the recombinant strains. When serum containing FomA-specific antibodies was incubated with the F. nucleatum in vitro, the number of Porphyromonas gingivalis cells that coaggregated with the F. nucleatum cells was significantly reduced. Furthermore, a mouse gum abscess model was successfully generated, and the range of gingival abscesses in the immune mice was relatively limited compared with the control group. The level of IL-1β in the serum and local gum tissues of the immune mice was consistently lower than in the control group. Our findings indicated that oral administration of the recombinant L. acidophilus reduced the risk of periodontal infection with P. gingivalis and F. nucleatum.

  2. Potential for tyndalized Lactobacillus acidophilus as an effective component in moisturizing skin and anti-wrinkle products.

    PubMed

    Im, A-Rang; Kim, Hui Seong; Hyun, Jin Won; Chae, Sungwook

    2016-08-01

    It is widely accepted that ultraviolet (UV) irradiation induces skin damage. In the present study, a UVB-induced hairless mouse model of skin photoaging was developed to determine whether tyndalized Lactobacillus acidophilus was able to significantly enhance the repair of photodamaged skin. To evaluate the effects of tyndalized L. acidophilus on UVB-induced skin-wrinkle formation in vivo , HR-1 hairless male mice were exposed to UVB radiation and orally administered tyndalized L. acidophilus . Compared with the control group, the UVB irradiation mice displayed a significant increase in transepidermal water loss and a reduction in skin hydration. In mice with UVB-induced photodamage, the effacement of the fine wrinkles by tyndalized L. acidophilus was correlated with dermal collagen synthesis, accompanied by histological changes. Furthermore, western blotting was performed to investigate the protein expression levels of matrix metalloproteinases (MMPs) and mitogen-activated protein kinase. Notably, orally administered tyndalized L. acidophilus reduced the expression levels of MMP-1 and MMP-9. Based upon the aforementioned results, it was determined that tyndalized L. acidophilus effectively inhibited the wrinkle formation induced by UVB irradiation, and that this may be attributed to the downregulation of MMPs. Therefore, tyndalized L. acidophilus may be considered a potential agent for preventing skin photoaging and wrinkle formation.

  3. Lactobacillus acidophilus Improves Intestinal Inflammation in an Acute Colitis Mouse Model by Regulation of Th17 and Treg Cell Balance and Fibrosis Development.

    PubMed

    Park, Jin-Sil; Choi, Jeong Won; Jhun, JooYeon; Kwon, Ji Ye; Lee, Bo-In; Yang, Chul Woo; Park, Sung-Hwan; Cho, Mi-La

    2018-03-01

    Disruption of the balance among the microbiota, epithelial cells, and resident immune cells in the intestine is involved in the pathogenesis of inflammatory bowel disease (IBD). Probiotics exert protective effects against IBD, and probiotic commensal Lactobacillus species are common inhabitants of the natural microbiota, especially in the gut. To investigate the effects of Lactobacillus acidophilus on the development of IBD, L. acidophilus was administered orally in mice with dextran sodium sulfate (DSS)-induced colitis. DSS-induced damage and the therapeutic effect of L. acidophilus were investigated. Treatment with L. acidophilus attenuated the severity of DSS-induced colitis. Specifically, it suppressed proinflammatory cytokines such as interleukin (IL)-6, tumor necrosis factor-α, IL-1β, and IL-17 in the colon tissues, which are produced by T helper (Th) 17 cells. Moreover, in vitro L. acidophilus treatment directly induced T regulatory (Treg) cells and the production of IL-10, whereas the production of IL-17 was suppressed in splenocytes. In addition, we found that L. acidophilus treatment decreased the levels of α-smooth muscle actin, a marker of activated myofibroblasts, and type I collagen compared with control mice. These results suggest that L. acidophilus may be a novel treatment for IBD by modulating the balance between Th17 and Treg cells, as well as fibrosis development.

  4. Activities of free and encapsulated Lactobacillus acidophilus LA5 or Lactobacillus casei 01 in processed longan juices on exposure to simulated gastrointestinal tract.

    PubMed

    Chaikham, Pittaya; Apichartsrangkoon, Arunee; Worametrachanon, Srivilai; Supraditareporn, Wissanee; Chokiatirote, Ekachai; Van der Wiele, Tom

    2013-07-01

    Fruit drinks containing probiotics are gaining interest in the global marketplace. For example, longan juice, containing carbohydrate and various bioactive components, is a potentially health-promoting beverage as well as probiotic carrier for human consumption. In this study, high-pressure and thermal processes were applied to eliminate competitive micro-organisms in longan juice prior to the addition of Lactobacillus acidophilus LA5 or Lactobacillus casei 01. The activities of these probiotics in a simulated gastrointestinal tract were also investigated. Encapsulated probiotics could survive in the acidic environment of the stomach and small intestine, while the free cells were completely eliminated. In the colon experiment, the influence of encapsulated L. casei 01 on colon lactobacilli was significantly greater than that of encapsulated L. acidophilus LA5. Both encapsulated probiotics suspended in processed longan juices led to extensive increases in the formation of lactic acid and short-chain fatty acids (SCFA). Acetate was the major SCFA produced by colon bacteria, followed by propionate and butyrate. The discernible clear zone suggested that L. casei 01 provided greater antibacterial activity than L. acidophilus LA5. Both encapsulated probiotics along with processed longan juice led to significant increases in colon lactobacilli, lactic acid and SCFA formation. © 2012 Society of Chemical Industry.

  5. Surface layer protein from Lactobacillus acidophilus NCFM inhibit intestinal pathogen-induced apoptosis in HT-29 cells.

    PubMed

    Meng, Jun; Zhang, Qiu-Xiang; Lu, Rong-Rong

    2017-03-01

    Intestinal pathogens have been proposed to adhere to epithelial cells and cause apoptosis. This study was to investigate the inhibitory effects of surface layer protein (SLP, 46kDa) from Lactobacillus acidophilus NCFM on Escherichia coli and Salmonella-induced apoptosis in HT-29 cells and the mechanism of the inhibition was also studied. The SLP could alleviate the chromatin condensation caused by intestinal pathogens as observed under fluorescent microscope. Flow cytometry analysis showed that the SLP decreased E. coli and Salmonella-induced apoptosis by 46% and 48%, respectively. The SLP could also inhibit the mitochondrial membrane potential reduction and Ca 2+ level increase in HT-29 cells. Furthermore, the activation of caspase-9 and caspase-3 induced by E. coli and Salmonella was significantly decreased by the addition of SLP. These results suggested that L. acidophilus NCFM SLP could protect HT-29 cells against intestinal pathogen-induced apoptosis through a mitochondria-mediated pathway. These findings may reveal a new method for the treatment of intestinal infection and provide a theoretical basis for the practical application of SLP in food, biological and pharmaceutical fields. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Lactobacillus acidophilus alleviates the inflammatory response to enterotoxigenic Escherichia coli K88 via inhibition of the NF-κB and p38 mitogen-activated protein kinase signaling pathways in piglets.

    PubMed

    Li, Haihua; Zhang, Lei; Chen, Longbin; Zhu, Qi; Wang, Wenjie; Qiao, Jiayun

    2016-11-10

    A newly isolated L. acidophilus strain has been reported to have potential anti-inflammatory activities against lipopolysaccharide (LPS) challenge in piglet, while the details of the related inflammatory responses are limited. Here we aimed to analysis the ability of L. acidophilus to regulate inflammatory responses and to elucidate the mechanisms involved in its anti-inflammatory activity. The ETEC (enterotoxigenic Escherichia coli) K88-induced up-regulations of IL-1β, IL-8 and TNF-α were obviously inhibited by L. acidophilus while IL-10 was significantly increased. Moreover, L. acidophilus down-regulated pattern recognition receptors TLR (Toll-like receptor) 2 and TLR4 expression in both spleen and mesenteric lymph nodes of ETEC-challenged piglets, in accompanied with the reduced phosphorylation levels of nuclear factor kappa B (NF-κB) p65 and mitogen-activated protein kinase (MAPK) p38 as well in spleen of ETEC-infected piglets. Furthermore, L.acidophilus significantly increased the expression of the negative regulators of TLRs signaling, including Tollip, IRAK-M, A20 and Bcl-3 in spleen of ETEC-challenged piglets. Our findings suggested that L. acidophilus regulated inflammatory response to ETEC via impairing both NF-κB and MAPK signaling pathways in piglets.

  7. The Effect of Lactobacillus acidophilus PTCC 1643 on Cultured Intestinal Epithelial Cells Infected with Salmonella enterica serovar Enteritidis

    PubMed Central

    Moshiri, Mona; Dallal, Mohammad Mehdi Soltan; Rezaei, Farhad; Douraghi, Masoumeh; Sharifi, Laleh; Noroozbabaei, Zahra; Gholami, Mehrdad; Mirshafiey, Abbas

    2017-01-01

    Objectives Gastrointestinal disorders caused by Salmonella enterica serovar Enteritidis (SesE) are a significant health problem around the globe. Probiotic bacteria have been shown to have positive effects on the immune responses. Lactobacillus acidophilus was examined for its capability to influence the innate immune response of HT29 intestinal epithelial cells towards SesE. The purpose of this work was to assess the effect of L. acidophilus PTCC 1643 on cultured intestinal epithelial cells infected with SesE. Methods HT29 cells were cultured in Roswell Park Memorial Institute medium supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. The cells were treated with L. acidophilus PTCC 1643 after or before challenge with SesE. At 2 and 4 hours post-infection, we measured changes in the expression levels of TLR2 and TLR4 via real-time polymerase chain reaction. Results Treatment with L. acidophilus inhibited SesE-induced increases in TLR2 and TLR4 expression in the infected HT29 cells. Moreover, the expression of TLR2 and TLR4 in cells that were pretreated with L. acidophilus and then infected with SesE was significantly higher than that in cells infected with SesE without pretreatment. Taken together, the results indicated that L. acidophilus had an anti-inflammatory effect and modulated the innate immune response to SesE by influencing TLR2 and TLR4 expression. Conclusion Our findings suggested that L. acidophilus PTCC 1643 was able to suppress inflammation caused by SesE infection in HT29 cells and reduce TLR2 and TLR4 expression. Additional in vivo and in vitro studies are required to further elucidate the mechanisms underlying this anti-inflammatory effect. PMID:28443224

  8. The Effect of Lactobacillus acidophilus PTCC 1643 on Cultured Intestinal Epithelial Cells Infected with Salmonella enterica serovar Enteritidis.

    PubMed

    Moshiri, Mona; Dallal, Mohammad Mehdi Soltan; Rezaei, Farhad; Douraghi, Masoumeh; Sharifi, Laleh; Noroozbabaei, Zahra; Gholami, Mehrdad; Mirshafiey, Abbas

    2017-02-01

    Gastrointestinal disorders caused by Salmonella enterica serovar Enteritidis ( Se sE) are a significant health problem around the globe. Probiotic bacteria have been shown to have positive effects on the immune responses. Lactobacillus acidophilus was examined for its capability to influence the innate immune response of HT29 intestinal epithelial cells towards Se sE. The purpose of this work was to assess the effect of L. acidophilus PTCC 1643 on cultured intestinal epithelial cells infected with Se sE. HT29 cells were cultured in Roswell Park Memorial Institute medium supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. The cells were treated with L. acidophilus PTCC 1643 after or before challenge with Se sE. At 2 and 4 hours post-infection, we measured changes in the expression levels of TLR2 and TLR4 via real-time polymerase chain reaction. Treatment with L. acidophilus inhibited Se sE-induced increases in TLR2 and TLR4 expression in the infected HT29 cells. Moreover, the expression of TLR2 and TLR4 in cells that were pretreated with L. acidophilus and then infected with Se sE was significantly higher than that in cells infected with Se sE without pretreatment. Taken together, the results indicated that L. acidophilus had an anti-inflammatory effect and modulated the innate immune response to Se sE by influencing TLR2 and TLR4 expression. Our findings suggested that L. acidophilus PTCC 1643 was able to suppress inflammation caused by Se sE infection in HT29 cells and reduce TLR2 and TLR4 expression. Additional in vivo and in vitro studies are required to further elucidate the mechanisms underlying this anti-inflammatory effect.

  9. Deletion of Lipoteichoic Acid Synthase Impacts Expression of Genes Encoding Cell Surface Proteins in Lactobacillus acidophilus

    PubMed Central

    Selle, Kurt; Goh, Yong J.; Johnson, Brant R.; O’Flaherty, Sarah; Andersen, Joakim M.; Barrangou, Rodolphe; Klaenhammer, Todd R.

    2017-01-01

    Lactobacillus acidophilus NCFM is a well-characterized probiotic microorganism, supported by a decade of genomic and functional phenotypic investigations. L. acidophilus deficient in lipoteichoic acid (LTA), a major immunostimulant in Gram-positive bacteria, has been shown to shift immune system responses in animal disease models. However, the pleiotropic effects of removing LTA from the cell surface in lactobacilli are unknown. In this study, we surveyed the global transcriptional and extracellular protein profiles of two strains of L. acidophilus deficient in LTA. Twenty-four differentially expressed genes specific to the LTA-deficient strains were identified, including a predicted heavy metal resistance operon and several putative peptidoglycan hydrolases. Cell morphology and manganese sensitivity phenotypes were assessed in relation to the putative functions of differentially expressed genes. LTA-deficient L. acidophilus exhibited elongated cellular morphology and their growth was severely inhibited by elevated manganese concentrations. Exoproteomic surveys revealed distinct changes in the composition and relative abundances of several extracellular proteins and showed a bias of intracellular proteins in LTA-deficient strains of L. acidophilus. Taken together, these results elucidate the impact of ltaS deletion on the transcriptome and extracellular proteins of L. acidophilus, suggesting roles of LTA in cell morphology and ion homeostasis as a structural component of the Gram positive cell wall. PMID:28443071

  10. Deletion of Lipoteichoic Acid Synthase Impacts Expression of Genes Encoding Cell Surface Proteins in Lactobacillus acidophilus.

    PubMed

    Selle, Kurt; Goh, Yong J; Johnson, Brant R; O'Flaherty, Sarah; Andersen, Joakim M; Barrangou, Rodolphe; Klaenhammer, Todd R

    2017-01-01

    Lactobacillus acidophilus NCFM is a well-characterized probiotic microorganism, supported by a decade of genomic and functional phenotypic investigations. L. acidophilus deficient in lipoteichoic acid (LTA), a major immunostimulant in Gram-positive bacteria, has been shown to shift immune system responses in animal disease models. However, the pleiotropic effects of removing LTA from the cell surface in lactobacilli are unknown. In this study, we surveyed the global transcriptional and extracellular protein profiles of two strains of L. acidophilus deficient in LTA. Twenty-four differentially expressed genes specific to the LTA-deficient strains were identified, including a predicted heavy metal resistance operon and several putative peptidoglycan hydrolases. Cell morphology and manganese sensitivity phenotypes were assessed in relation to the putative functions of differentially expressed genes. LTA-deficient L. acidophilus exhibited elongated cellular morphology and their growth was severely inhibited by elevated manganese concentrations. Exoproteomic surveys revealed distinct changes in the composition and relative abundances of several extracellular proteins and showed a bias of intracellular proteins in LTA-deficient strains of L. acidophilus . Taken together, these results elucidate the impact of ltaS deletion on the transcriptome and extracellular proteins of L. acidophilus , suggesting roles of LTA in cell morphology and ion homeostasis as a structural component of the Gram positive cell wall.

  11. Regulatory role of Lactobacillus acidophilus on inflammation and gastric dysmotility in intestinal mucositis induced by 5-fluorouracil in mice.

    PubMed

    Justino, Priscilla F C; Melo, Luis F M; Nogueira, Andre F; Morais, Cecila M; Mendes, Walber O; Franco, Alvaro X; Souza, Emmanuel P; Ribeiro, Ronaldo A; Souza, Marcellus H L P; Soares, Pedro Marcos Gomes

    2015-03-01

    Lactobacillus acidophilus is widely used for gastrointestinal disorders, but its role in inflammatory conditions like in chemotherapy-induced mucositis is unclear. Here, we report the effect of L. acidophilus on 5-fluorouracil-induced (5-FU) intestinal mucositis in mice. Mice weighing 25-30 g (n = 8) were separated into three groups, saline, 5-FU, and 5-FU + L. acidophilus (5-FU-La) (16 × 10(9) CFU/kg). In the 5-FU-La group, L. acidophilus was administered concomitantly with 5-FU on the first day and alone for two additional days. Three days after the last administration of L. acidophilus, the animals were euthanized and the jejunum and ileum were removed for histopathological assessment and for evaluation of levels of myeloperoxidase activity, sulfhydryl groups, nitrite, and cytokines (TNF-α, IL-1β, CXCL-1, and IL-10). In addition, we investigated gastric emptying using spectrophotometry after feeding a 1.5-ml test meal by gavage and euthanasia. Data were submitted to ANOVA and Bonferroni's test, with the level of significance at p < 0.05. Intestinal mucositis induced by 5-FU significantly (p < 0.05) reduced the villus height-crypt depth ratio and GSH concentration and increased myeloperoxidase activity and the nitrite concentrations compared with the control group. Furthermore, 5-FU significantly (p < 0.05) increased cytokine (TNF-α, IL-1β, and CXCL-1) concentrations and decreased IL-10 concentrations compared with the control group. 5-FU also significantly (p < 0.05) delayed gastric emptying and gastrointestinal transit compared with the control group. All of these changes were significantly (p < 0.05) reversed by treatment with L. acidophilus. Lactobacillus acidophilus improves the inflammatory and functional aspects of intestinal mucositis induced by 5-FU.

  12. Poultry offal ensiled with lactobacillus acidophilus for growing and finishing swine diets.

    PubMed

    Tibbetts, G W; Seerley, R W; McCampbell, H C

    1987-01-01

    Poultry offal (heads, feet, viscera) from a broiler processing plant was ground and mixed with corn, dried molasses and a Lactobacillus acidophilus culture. Fermentation was shown to be effective in batch sizes of 3.8, 75.7 and 208.1 liters, as well as a 1,360-kg silo. It was generally complete at 72 h. In trial 1, rats were fed silage mixtures of 60:30:5:5, 45:45:5:5 and 30:60:5:5, offal, corn, molasses and inoculant, respectively. Rats did not gain as well when fed the silage diets (P less than .05) as when fed the basal diet; however, the ranking of silages was 45:45, 60:30 and 30:60, offal-to-corn ratio for rat daily gains and feed conversions. In trial 2, growing-finishing pigs were fed the 60:30:5:5 silage at rates of 0, 10, 20 and 30% of the diet. Average daily gains and feed-to-gain ratios were not affected by including offal silage at up to 20% of the diet, but 30% offal silage in diets resulted in depressed gains (P less than .05) and increased feed-to-gain ratios. Carcasses were not different among treatments for dressing percentage, length, average backfat and percentage of ham, loin, shoulder, lean cuts and primal cuts. Carcasses from pigs fed 20 and 30% offal silage had significantly darker and firmer loin-eyes than those from control fed pigs. Marbling was higher (P less than .05) in loin-eyes from pigs fed 30% offal silage as compared with those from pigs fed 0% offal silage.(ABSTRACT TRUNCATED AT 250 WORDS)

  13. Anaerobic sludge digestion with a biocatalytic additive. [Lactobacillus acidophilus

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ghosh, S.; Henry, M.P.; Fedde, P.A.

    1982-01-01

    Aimed at improving the process operating characteristics of anaerobic digestion for sludge stabilization and SNG production, this study evaluates the effects of a lactobacillus additive under normal, variable, and overload conditions. This whey fermentation product of an acid-tolerant strain of L. acidophilus fortified with CoCO/sub 3/, (NH/sub 4/)/sub 2/HPO/sub 4/, ferrous lactate, and lactic acid provides growth factors, metabolic intermediates, and enzymes needed for substrate degradation and cellular synthesis. Data indicate that the biochemical additive increases methane yield, gas production rate, and volatile solids reduction; decreases volatile acids accumulation; enhances the digester buffer capacity; and improves the fertilizer value andmore » dewatering characteristics of the digested residue. Digester capacities could be potentially doubled when the feed is so treated. Results of field tests with six full-scale digesters confirm observations made with bench-scale digesters.« less

  14. Inhibitory effects of antiseptic mouthrinses on Streptococcus mutans, Streptococcus sanguinis and Lactobacillus acidophilus.

    PubMed

    Evans, A; Leishman, S J; Walsh, L J; Seow, W K

    2015-06-01

    Oral antiseptics are valuable in controlling oral infections caused by cariogenic bacteria. The aim of this study was to investigate the effects of mouthrinses and pure antiseptic compounds on Streptococcus mutans and non-mutans bacteria (Streptococcus sanguinis and Lactobacillus acidophilus). The agar diffusion assay was employed to determine bacterial growth inhibition. Commercial mouthrinses containing chlorhexidine gluconate (0.2%), cetylpyridinium chloride (0.05%) and sodium fluoride (0.05%) produced statistically similar growth inhibition of S. mutans, S. sanguinis and L. acidophilus (with zones of inhibition ranging from 7.56 ± 0.52 mm to 7.39 ± 0.53 mm, 17.44 ± 0.94 mm to 18.31 ± 0.62 mm and 8.61 ± 1.43 to 8.67 ± 1.43 mm respectively, p > 0.05). The chlorhexidine mouthwash produced the greatest mean growth inhibition of S. sanguinis and S. mutans compared to all other mouthrinses tested (p < 0.01). The minimum concentrations at which inhibition against S. mutans could be detected were chlorhexidine gluconate at 0.005% (wt/vol), cetylpyridinium chloride 0.01% (wt/ vol), povidone iodine 10% (wt/vol) and sodium hypochlorite 0.5% (vol/vol). Chlorhexidine (0.01%), cetylpyridinium chloride (0.01%), povidone iodine (10%) and sodium hypochlorite (0.5%) are effective at inhibiting the growth of S. mutans, S. sanguinis and L. acidophilus. © 2015 Australian Dental Association.

  15. Lactobacillus acidophilus CL1285, Lactobacillus casei LBC80R and Lactobacillus rhamnosus CLR2 improve quality-of-life and IBS symptoms: a double-blind, randomised, placebo-controlled study.

    PubMed

    Preston, K; Krumian, R; Hattner, J; de Montigny, D; Stewart, M; Gaddam, S

    2018-06-11

    A combination of Lactobacillus acidophilus CL1285, Lactobacillus casei LBC80R and Lactobacillus rhamnosus CLR2 was compared to placebo for relief of symptoms of irritable bowel syndrome (IBS). A total of 113 subjects at 3 clinical sites were randomised in a 2:1 ratio and followed for 12 weeks. Subjects ingested either 2 capsules of active study product, containing 50×10 9 cfu of live organisms, or 2 placebo capsules daily. Endpoints included improvement in abdominal pain, days of pain, distention, stool consistency and frequency, quality of life (QOL), and adequate relief (AR) of IBS symptoms. IBS subtypes constipation (IBS-C), diarrhoea (IBS-D), and mixed (IBS-M) were evaluated separately; the effect of gender was also examined. For all efficacy endpoints improvement of 30% or more vs placebo was considered clinically significant. With the exception of pain intensity and AR, the endpoints demonstrated a therapeutic advantage of active over placebo for IBS symptoms in at least some subject subgroups. The IBS-D and female subgroups showed the largest and most consistent effects. Stool frequency and consistency were evaluated in the IBS-C and IBS-D subgroups, and improvement of active vs placebo was noted in both. QOL improvement was seen overall and in specific domains. Adverse events (AEs) were limited to 7 subjects; all were of mild or moderate intensity except one, severe cramping. Four AEs in the same subject in the placebo group were judged to be related to study product; these resolved by the end of study. There were no serious AEs.

  16. Transcriptional Analysis of Prebiotic Uptake and Catabolism by Lactobacillus acidophilus NCFM

    PubMed Central

    Andersen, Joakim Mark; Barrangou, Rodolphe; Hachem, Maher Abou; Lahtinen, Sampo J.; Goh, Yong-Jun; Svensson, Birte; Klaenhammer, Todd R.

    2012-01-01

    The human gastrointestinal tract can be positively modulated by dietary supplementation of probiotic bacteria in combination with prebiotic carbohydrates. Here differential transcriptomics and functional genomics were used to identify genes in Lactobacillus acidophilus NCFM involved in the uptake and catabolism of 11 potential prebiotic compounds consisting of α- and β- linked galactosides and glucosides. These oligosaccharides induced genes encoding phosphoenolpyruvate-dependent sugar phosphotransferase systems (PTS), galactoside pentose hexuronide (GPH) permease, and ATP-binding cassette (ABC) transporters. PTS systems were upregulated primarily by di- and tri-saccharides such as cellobiose, isomaltose, isomaltulose, panose and gentiobiose, while ABC transporters were upregulated by raffinose, Polydextrose, and stachyose. A single GPH transporter was induced by lactitol and galactooligosaccharides (GOS). The various transporters were associated with a number of glycoside hydrolases from families 1, 2, 4, 13, 32, 36, 42, and 65, involved in the catabolism of various α- and β-linked glucosides and galactosides. Further subfamily specialization was also observed for different PTS-associated GH1 6-phospho-β-glucosidases implicated in the catabolism of gentiobiose and cellobiose. These findings highlight the broad oligosaccharide metabolic repertoire of L. acidophilus NCFM and establish a platform for selection and screening of both probiotic bacteria and prebiotic compounds that may positively influence the gastrointestinal microbiota. PMID:23028535

  17. Viability of Lactobacillus acidophilus in synbiotic guava mousses and its survival under in vitro simulated gastrointestinal conditions.

    PubMed

    Buriti, Flávia C A; Castro, Inar A; Saad, Susana M I

    2010-02-28

    The effects of refrigeration, freezing and substitution of milk fat by inulin and whey protein concentrate (WPC) on Lactobacillus acidophilus La-5 viability and resistance to gastric and enteric simulated conditions in synbiotic guava mousses effects were investigated. Refrigerated mousses supplemented with WPC presented the best probiotic viability, ranging from 7.77 to 6.24 log cfu/g during 28 days of storage. The highest probiotic populations, above 7.45 log cfu/g, were observed for all frozen mousses during 112 days of storage. Decreased L. acidophilus survival during the in vitro gastrointestinal simulation was observed both for refrigerated and frozen mousses. Nonetheless, for the refrigerated mousses, the addition of inulin enhanced the probiotic survival during the in vitro assays in the first week of storage. L. acidophilus survival in simulated gastrointestinal fluids was also improved through freezing. The frozen storage may be used to provide increased shelf-life for synbiotic guava mousses. Even though the protective effect of inulin and WPC on the probiotic microorganism tested was shown to be more specific for the refrigerated products, the partial replacement of milk fat by these ingredients may also help, as it improves the nutritional value of mousses in both storage conditions. (c) 2009 Elsevier B.V. All rights reserved.

  18. Lactobacillus acidophilus suppresses intestinal inflammation by inhibiting endoplasmic reticulum stress.

    PubMed

    Kim, Da Hye; Kim, Soochan; Lee, Jin Ha; Kim, Jae Hyeon; Che, Xiumei; Ma, Hyun Woo; Seo, Dong Hyuk; Kim, Tae Il; Kim, Won Ho; Kim, Seung Won; Cheon, Jae Hee

    2018-06-22

    Nuclear factor kappa B (NF-κB) activation and endoplasmic reticulum (ER) stress signaling play significant roles in the pathogenesis of inflammatory bowel disease (IBD). Thus, we evaluated whether new therapeutic probiotics have anti-colitic effects and we investigated their mechanisms related to NF-κB and ER-stress pathways. Luciferase, nitric oxide (NO), and cytokine assays using HT-29 or RAW264.7 cells were conducted. Mouse colitis was induced using dextran sulfate sodium (DSS) and confirmed by disease activity index and histology. Macrophages and T-cell subsets in isolated peritoneal cavity cells (PCCs) and splenocytes were analyzed by flow cytometry. Gene and cytokine expression profiles were determined using RT-PCR. Lactobacillus acidophilus (LA1) and Pediococcus pentosaceus inhibited NO production in RAW264.7 cells, but only LA1 inhibited Tnfa and induced Il10 expression. LA1 increased the life span of DSS-treated mice and attenuated the severity of colitis by inducing M2 macrophages in PCCs and Th2 and Treg cells in splenocytes. The restoration of goblet cells in the colon was accompanied by the induction of Il10 expression and the suppression of proinflammatory cytokines. Additionally, we found that LA1 exerts an anti-colitic effect by improving ER stress in HT-29 cells as well as in vivo. We showed that LA1 significantly interferes with ER stress and suppresses NF-κB activation. Our findings suggest that LA1 can be used as a potent immunomodulator in IBD treatment and the regulation of ER stress may have significant implications in treating IBD. This article is protected by copyright. All rights reserved.

  19. Probiotic Yogurt Culture Bifidobacterium Animalis Subsp. Lactis BB-12 and Lactobacillus Acidophilus LA-5 Modulate the Cytokine Secretion by Peripheral Blood Mononuclear Cells from Patients with Ulcerative Colitis.

    PubMed

    Sheikhi, A; Shakerian, M; Giti, H; Baghaeifar, M; Jafarzadeh, A; Ghaed, V; Heibor, M R; Baharifar, N; Dadafarin, Z; Bashirpour, G

    2016-06-01

    There are some evidences for the immunomodulation disorders in the response to intestinal microbiota in inflammatory bowel disease. Yogurt is a fermented milk product made with a starter culture consisting of different probiotics which could be colonized in intestine. However, the role of probiotics in the aetiopathogenesis of ulcerative colitis (UC) has not been clarified. To determine how the immune system responds to these bacteria this study was planned. Bifidobacterium lactis BB-12 (B. lactis) and Lactobacillus acidophilus LA-5 (L. acidophilus) were cultivated on MRS broth. PBMCs of 36 UC patients were separated by Ficoll-Hypaque centrifugation and co-cultured with different concentrations of UV killed bacteria in RPMI-1 640 plus 10% FCS for 48/72 h. IL-10, TGF-β, IFN-γ and TNF-α were measured in supernatant of PBMCs by ELISA. Both bacteria significantly augmented IL-10, TGF-β, IFN-γ and TNF-α compared to control (p<0.001). The secretion levels of IL-10 and TGF-β by B. lactis- compared to L. acidophilus-stimulated PBMCs were significantly higher (p<0.05, p<0.01 respectively). The secretion levels of TNF-α and IFN-γ by PBMCs after 72 h were significantly lower compared to 48 h stimulation by B. lactis (p<0.001, p<0.035 respectively). These data show that both probiotics may trigger the pro- and anti-inflammatory immune response of UC patients. It seems that IL-10/TGF-β uprising by B. lactis could be the reason of TNF-α/IFN-γ reduction. Therefore albeit B. lactis still stimulates the effector Th cells but because of more stimulatory effect on Tregs, it could be a good potential therapeutic candidate for further investigation. © Georg Thieme Verlag KG Stuttgart · New York.

  20. A functional glycogen biosynthesis pathway in Lactobacillus acidophilus: expression and analysis of the glg operon

    PubMed Central

    Goh, Yong Jun; Klaenhammer, Todd R

    2013-01-01

    Glycogen metabolism contributes to energy storage and various physiological functions in some prokaryotes, including colonization persistence. A role for glycogen metabolism is proposed on the survival and fitness of Lactobacillus acidophilus, a probiotic microbe, in the human gastrointestinal environment. L. acidophilus NCFM possesses a glycogen metabolism (glg) operon consisting of glgBCDAP-amy-pgm genes. Expression of the glg operon and glycogen accumulation were carbon source- and growth phase-dependent, and were repressed by glucose. The highest intracellular glycogen content was observed in early log-phase cells grown on trehalose, which was followed by a drastic decrease of glycogen content prior to entering stationary phase. In raffinose-grown cells, however, glycogen accumulation gradually declined following early log phase and was maintained at stable levels throughout stationary phase. Raffinose also induced an overall higher temporal glg expression throughout growth compared with trehalose. Isogenic ΔglgA (glycogen synthase) and ΔglgB (glycogen-branching enzyme) mutants are glycogen-deficient and exhibited growth defects on raffinose. The latter observation suggests a reciprocal relationship between glycogen synthesis and raffinose metabolism. Deletion of glgB or glgP (glycogen phosphorylase) resulted in defective growth and increased bile sensitivity. The data indicate that glycogen metabolism is involved in growth maintenance, bile tolerance and complex carbohydrate utilization in L. acidophilus. PMID:23879596

  1. Assessment of in vitro oxalate degradation by Lactobacillus species cultured from veterinary probiotics.

    PubMed

    Cho, Jenny G; Gebhart, Connie J; Furrow, Eva; Lulich, Jody P

    2015-09-01

    To culture Lactobacillus spp from veterinary probiotics and measure their in vitro oxalate-degrading capacity. 2 commercial veterinary probiotics containing Lactobacillus spp. Lactobacillus spp were cultured anaerobically on selective deMan, Rogosa, Sharpe agar medium and subcultured for speciation by 16S rDNA gene sequencing. Isolates were inoculated into broth containing sodium oxalate (5 mg/L) and incubated anaerobically for 72 hours. An oxalate-degrading isolate of Lactobacillus acidophilus (American Type Culture Collection [ATCC] 53544) was the positive control sample; sterile broth containing a known quantity of sodium oxalate was the negative control sample. Oxalate concentrations were detected with ion chromatography. Oxalate degradation was assessed with Dunnett tests to detect differences in mean oxalate concentration for each isolate, compared with results for the negative control. Lactobacillus acidophilus, Lactobacillus plantarum, and Lactobacillus casei or Lactobacillus zeae (too closely related to differentiate) were isolated from probiotic 1, and L plantarum was isolated from probiotic 2. Sequencing of the 16S rDNA gene confirmed 100% homology to type species. Lactobacillus acidophilus (ATCC 53544) and L acidophilus from probiotic 1 significantly decreased oxalate concentrations by 85.3 and 161.9 mg/L, respectively. Lactobacillus plantarum from probiotics 1 and 2 significantly increased oxalate concentrations by 56.1 and 36.1 mg/L, respectively. Lactobacillus casei did not alter oxalate concentrations. Lactobacillus acidophilus isolates significantly reduced oxalate concentrations. In vivo studies are needed to determine whether probiotics containing L acidophilus decrease urine oxalate concentrations and reduce risk of urolith recurrence in dogs with a history of calcium oxalate urolithiasis.

  2. Design and evaluation of an oral multiparticulate system for dual delivery of amoxicillin and Lactobacillus acidophilus.

    PubMed

    Govender, Mershen; Choonara, Yahya E; van Vuuren, Sandy; Kumar, Pradeep; du Toit, Lisa C; Pillay, Viness

    2016-09-01

    A delayed-release dual delivery system for amoxicillin and the probiotic Lactobacillus acidophilus was developed and evaluated. Statistical optimization of a cross-linked denatured ovalbumin protective matrix was first synthesized using a Box-Behnken experimental design prior to encapsulation with glyceryl monostereate. The encapsulated ovalbumin matrix was thereafter incorporated with amoxicillin in a gastro-resistant capsule. In vitro characterization and stability analysis of the ovalbumin and encapsulated components were also performed Results: Protection of L. acidophilus probiotic against the bactericidal effects of amoxicillin within the dual formulation was determined. The dual formulation in this study proved effective and provides insight into current microbiome research to identify, classify and use functional healthy bacteria to develop novel probiotic delivery technologies.

  3. Characteristic odor components of volatile oil from the cultivation medium of Lactobacillus acidophilus.

    PubMed

    Ono, Toshirou; Yonejima, Yasunori; Ikeda, Atsushi; Kashima, Yusei; Nakaya, Satoshi; Miyazawa, Mitsuo

    2014-01-01

    Volatile oils obtained from both the liquid medium after incubation (MAI) and liquid medium before incubation (MBI) in the cultivation process of Lactobacillus acidophilus were isolated by hydrodistillation (HD) and analyzed to investigate the utility of the liquid waste. The composition of the volatile oils was analyzed by capillary gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). In total, 46 and 19 compounds were detected in the volatile oils from MAI (MAI oil) and MBI (MBI oil), respectively. The principle components of MAI oil were fatty acids, including pentanoic acid (12.75%), heptanoic acid (14.05%), and nonanoic acid (14.04%). The important aroma-active compounds in the oils were detected by GC-MS/Olfactometry (GC-O), and their intensity of aroma were measured by aroma extraction dilution analysis (AEDA). Pyrazines were determined as key aroma components; in particular, 2-ethyl-5-methylpyrazine was the most primary aroma-active compound in MAI oil. In addition, as the characteristic aroma-active compounds, 3-(methylthio)-propanal, trimethylpyrazine, and pentanoic acid were also detected in MAI oil. These results imply that the waste medium after incubation of L. acidophilus may be utilized as a source of volatile oils.

  4. In vitro assessment of photocatalytic titanium oxide surface modified stainless steel orthodontic brackets for antiadherent and antibacterial properties against Lactobacillus acidophilus.

    PubMed

    Shah, Alok Girish; Shetty, Pradeep Chandra; Ramachandra, C S; Bhat, N Sham; Laxmikanth, S M

    2011-11-01

    To assess the antiadherent and antibacterial properties of surface modified stainless steel orthodontic brackets with photocatalytic titanium oxide (TiO(2)) against Lactobacillus acidophilus. This study was done on 120 specimens of stainless steel preadjusted edgewise appliance (PEA) orthodontic brackets. The specimens were divided into four test groups. Each group consisted of 30 specimens. Groups containing uncoated brackets acted as a control group for their respective experimental group containing coated brackets. Surface modification of brackets was carried out by the radiofrequency (RF) magnetron sputtering method with photocatalytic TiO(2). Brackets then were subjected to microbiological tests for assessment of the antiadherent and antibacterial properties of photocatalytic TiO(2) coating against L acidophilus. Orthodontic brackets coated with photocatalytic TiO(2) showed an antiadherent effect against L acidophilus compared with uncoated brackets. The bacterial mass that was bound to the TiO(2)-coated brackets was less when compared with the uncoated brackets. Furthermore, TiO(2)-coated brackets had a bactericidal effect on L acidophilus, which causes dental caries. Surface modification of orthodontic brackets with photocatalytic TiO(2) can be used to prevent the accumulation of dental plaque and the development of dental caries during orthodontic treatment.

  5. The potential risks of probiotics among HIV-infected persons: Bacteraemia due to Lactobacillus acidophilus and review of the literature.

    PubMed

    Haghighat, Leila; Crum-Cianflone, Nancy F

    2016-11-01

    Lactobacillus sp. are commensal organisms that are increasingly reported to cause invasive infections among immunosuppressed persons. However, few data exist regarding the occurrence and risk factors of these infections among HIV-infected persons. Further, the safety of products that contain lactobacilli (e.g. probiotics) in certain populations, including those with HIV/AIDS, is unclear. We report a case of Lactobacillus acidophilus bacteraemia in a patient with AIDS temporally related to excessive consumption of probiotic-enriched yogurt, and provide a comprehensive review of the literature of Lactobacillus sp. infections among HIV-infected persons. © The Author(s) 2015.

  6. Supplementation with a Lactobacillus acidophilus fermentation product alters the metabolic response following a lipopolysaccharide challenge in weaned pigs

    USDA-ARS?s Scientific Manuscript database

    This study was designed to determine if feeding a Lactobacillus acidophilus fermentation product to weaned pigs would alter the metabolic response following a lipopolysaccharide (LPS) challenge. Pigs (n=30; 6.4+/-0.1 kg BW) were housed individually with ad libitum access to feed and water. Pigs were...

  7. Supplementation of Lactobacillus acidophilus fermentation product can attenuate the acute phase response following a lipopolysaccharide challenge in pigs.

    USDA-ARS?s Scientific Manuscript database

    This study was designed to determine if feeding a Lactobacillus acidophilus fermentation product to weaned pigs would reduce stress and acute phase responses (APR) following a lipopolysaccharide (LPS) challenge. Pigs (n=30; 6.4±0.1 kilograms body weight) were housed individually in pens with ad libi...

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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 ...

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

    USDA-ARS?s Scientific Manuscript database

    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 ...

  11. A Decade of Experience in Primary Prevention of Clostridium difficile Infection at a Community Hospital Using the Probiotic Combination Lactobacillus acidophilus CL1285, Lactobacillus casei LBC80R, and Lactobacillus rhamnosus CLR2 (Bio-K+).

    PubMed

    Maziade, Pierre-Jean; Pereira, Pascale; Goldstein, Ellie J C

    2015-05-15

    In August 2003, the 284-bed community hospital Pierre-Le Gardeur (PLGH) in Quebec experienced a major outbreak associated with the Clostridium difficile NAP1/027/BI strain. Augmented standard preventive measures (SPMs) were not able to control this outbreak. It was decided in February 2004 to give to every adult inpatient on antibiotics, without any exclusion, a probiotic (Bio-K+: Lactobacillus acidophilus CL1285, Lactobacillus casei LBC80R, and Lactobacillus rhamnosus CLR2) within 12 hours of the antibiotic prescription. Augmented SPMs were continued. The use of the probiotic in addition to SPMs was associated with a marked reduction of C. difficile infection (CDI). During the 10 years of observation, 44 835 inpatients received Bio-K+, and the CDI rate at PLGH declined from 18.0 cases per 10,000 patient-days and remained at low mean levels of 2.3 cases per 10,000 patient-days. Additionally, 10-year data collected by the Ministry of Health in Quebec comparing the CDI rate between Quebec hospitals showed that CDI rates at PLGH were consistently and continuously lower compared with those at similar hospitals. Blood cultures were monitored at PLGH for Lactobacillus bacteremia through the 10 years' experience, and no Lactobacillus bacteremias were detected. Despite the limitation of an observational study, we concluded that the probiotic Bio-K+ was safe and effective in decreasing our primary CDI rate. © The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  12. Safety evaluation of HOWARU® Restore (Lactobacillus acidophilus NCFM, Lactobacillus paracasei Lpc-37, Bifidobacterium animalis subsp. lactis Bl-04 and B. lactis Bi-07) for antibiotic resistance, genomic risk factors, and acute toxicity.

    PubMed

    Morovic, Wesley; Roper, Jason M; Smith, Amy B; Mukerji, Pushkor; Stahl, Buffy; Rae, Jessica Caverly; Ouwehand, Arthur C

    2017-12-01

    Although probiotic lactobacilli and bifidobacteria are generally considered safe by various regulatory agencies, safety properties, such as absence of transferable antibiotic resistance, must still be determined for each strain prior to market introduction as a probiotic. Safety requirements for probiotics vary regionally and evaluation methods are not standardized, therefore methodologies are often adopted from food ingredients or chemicals to assess microbial safety. Four individual probiotic strains, Lactobacillus acidophilus NCFM ® , Lactobacillus paracasei Lpc-37 ® , Bifidobacterium animalis subsp. lactis strains Bl-04 ® , and Bi-07 ® , and their combination (HOWARU ® Restore) were examined for antibiotic resistance by broth microdilution culture, toxin genes by PCR and genome mining, and acute oral toxicity in rats. Only B. lactis Bl-04 exhibited antibiotic resistance above a regulated threshold due to a tetW gene previously demonstrated to be non-transferable. Genomic mining did not reveal any bacterial toxin genes known to harm mammalian hosts in any of the strains. The rodent studies did not indicate any evidence of acute toxicity following a dose of 1.7-4.1 × 10 12  CFU/kg body weight. Considering a 100-fold safety margin, this corresponds to 1.2-2.8 × 10 12  CFU for a 70 kg human. Our findings demonstrate a comprehensive approach of in vitro, in silico, and in vivo safety testing for probiotics. Copyright © 2017 The Author(s). Published by Elsevier Ltd.. All rights reserved.

  13. Reversal in fatigued athletes of a defect in interferon gamma secretion after administration of Lactobacillus acidophilus.

    PubMed

    Clancy, R L; Gleeson, M; Cox, A; Callister, R; Dorrington, M; D'Este, C; Pang, G; Pyne, D; Fricker, P; Henriksson, A

    2006-04-01

    Fatigue and impaired performance in athletes is well recognised and has been loosely linked to "overtraining". Reduced concentration of IgA in the saliva and increased shedding of Epstein Barr virus (EBV) have been associated with intense training in elite athletes. To determine whether athletes presenting with fatigue and impaired performance had an immune defect relevant to defective containment of EBV infection, and whether a probiotic preparation (Lactobacillus acidophilus) shown to enhance mucosal immunity in animal models could reverse any detected abnormality. The fatigued athletes had clinical characteristics consistent with re-activation of EBV infection and significantly (p = 0.02) less secretion of interferon (IFN) gamma from blood CD4 positive T cells. After one month of daily capsules containing 2 x 10(10) colony forming units of L acidophilus, secretion of IFNgamma from T cells had increased significantly (p = 0.01) to levels found in healthy control athletes. A significant (p = 0.03) increase in salivary IFNgamma concentrations in healthy control athletes after the one month course of L acidophilus demonstrated in man the capacity for this probiotic to enhance the mucosal IFNgamma concentration. This is the first evidence of a T cell defect in fatigued athletes, and of its reversal following probiotic therapy.

  14. Development of a chromosome-plasmid balanced lethal system for Lactobacillus acidophilus with thyA gene as selective marker.

    PubMed

    Fu, X; Xu, J G

    2000-01-01

    A chromosome-plasmid balanced lethal gene delivery system for Lactobacillus acidophilus based on the thyA gene was developed. The selected L. acidophilus DOM La strain carries a mutated thyA gene and has an obligate requirement for thymidine. This strain can be used as a host for the constructed shuttle vector pFXL03, lacking antibiotic-resistant markers but having the wild-type thyA gene from L. casei which complements the thyA chromosomal mutation. The vector also contains the replicon region from plasmid pUC19 and that of the Lactococcus plasmid pWV01, which allows the transfer between Escherichia coli, L. casei and L. acidophilus. Eight unique restriction sites (i.e., PstI, HindIII, SphI, SalI, AccI, XbaI, KpnI and SacI) are available for cloning. After 40-time transfers in modified MRS medium, no plasmid loss was observed. The vector pFXL03 is potentially useful as a food-grade vaccine delivery system for L. acidophilus.

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

    USDA-ARS?s Scientific Manuscript database

    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...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  17. Comparable effects of low-intensity electromagnetic irradiation at the frequency of 51.8 and 53 GHz and antibiotic ceftazidime on Lactobacillus acidophilus growth and survival.

    PubMed

    Soghomonyan, Diana; Trchounian, Armen

    2013-01-01

    The effects of low-intensity electromagnetic irradiation (EMI) with the frequencies of 51.8 and 53 GHz on Lactobacillus acidophilus growth and survival were revealed. These effects were compared with antibacterial effects of antibiotic ceftazidime. Decrease in bacterial growth rate by EMI was comparable with the inhibitory effect of ceftazidime (minimal inhibitory concentration-16 μM) and no enhanced action was observed with combined effects of EMI and the antibiotic. However, EMI-enhanced antibiotic inhibitory effect on bacterial survival. The kinetics of the bacterial suspension oxidation-reduction potential up to 24 h of the growth was changed by EMI and ceftazidime. The changes were more strongly expressed by combined effects of EMI and antibiotic especially up to 12 h. Moreover, EMI did not change overall energy (glucose)-dependent H(+) efflux across the membrane but it increased N,N'-dicyclohexylcarbodiimide (DCCD)-inhibited H(+) efflux. In contrast, this EMI in combination with ceftazidime decreased DCCD-sensitive H(+) efflux. Low-intensity EMI had inhibitory effect on L. acidophilus bacterial growth and survival. The effect on bacterial survival was more significant in the combination with ceftazidime. The H(+)-translocating F 0 F 1-ATPase, for which DCCD is specific inhibitor, might be a target for EMI and ceftazidime. The revealed bactericide effects on L. acidophilus can be applied in biotechnology, food producing and safety technology.

  18. 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.

  19. Multivalent Chromosomal Expression of the Clostridium botulinum Serotype A Neurotoxin Heavy-Chain Antigen and the Bacillus anthracis Protective Antigen in Lactobacillus acidophilus.

    PubMed

    O'Flaherty, Sarah; Klaenhammer, Todd R

    2016-10-15

    -based expression systems since expressing antigens from a chromosomal location confers an advantage to the vaccine strains by eliminating the antibiotic maintenance required for plasmids and negates issues with plasmid instability that would result in loss of the antigen. Lactic acid bacteria, including Lactobacillus acidophilus, have shown potential for mucosal vaccine delivery, as L. acidophilus is bile and acid tolerant, allowing transit through the gastrointestinal tract where cells interact with host epithelial and immune cells, including dendritic cells. In this study, we successfully expressed C. botulinum and B. anthracis antigens in the probiotic L. acidophilus strain NCFM. Both antigens were highly expressed individually or in tandem from the chromosome of L. acidophilus. Copyright © 2016 O'Flaherty and Klaenhammer.

  20. Multivalent Chromosomal Expression of the Clostridium botulinum Serotype A Neurotoxin Heavy-Chain Antigen and the Bacillus anthracis Protective Antigen in Lactobacillus acidophilus

    PubMed Central

    Klaenhammer, Todd R.

    2016-01-01

    preferred over plasmid-based expression systems since expressing antigens from a chromosomal location confers an advantage to the vaccine strains by eliminating the antibiotic maintenance required for plasmids and negates issues with plasmid instability that would result in loss of the antigen. Lactic acid bacteria, including Lactobacillus acidophilus, have shown potential for mucosal vaccine delivery, as L. acidophilus is bile and acid tolerant, allowing transit through the gastrointestinal tract where cells interact with host epithelial and immune cells, including dendritic cells. In this study, we successfully expressed C. botulinum and B. anthracis antigens in the probiotic L. acidophilus strain NCFM. Both antigens were highly expressed individually or in tandem from the chromosome of L. acidophilus. PMID:27496774

  1. Construction and immunogenicity of the recombinant Lactobacillus acidophilus pMG36e-E0-LA-5 of bovine viral diarrhea virus.

    PubMed

    Zhao, Yuelan; Jiang, Lufeng; Liu, Teng; Wang, Min; Cao, Wenbo; Bao, Yongzhan; Qin, Jianhua

    2015-12-01

    Bovine viral diarrhea/mucosal disease (BVD/MD) is an infectious disease of cattle with a worldwide distribution, creating a substantial economic impact. It is caused by bovine viral diarrhea virus (BVDV). This research was conducted to construct the recombinant Lactobacillus acidophilus (L. acidophilus) pMG36e-E0-LA-5 of BVDV E0 gene and to test its immunogenicity and protective efficacy against BVDV infection in the mice model. The BVDV E0 gene was sub-cloned into the expression vector and then transformed into the L. acidophilus LA-5 strain by electroporation. The recombinant L. acidophilus pMG36e-E0-LA-5 was confirmed by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. The mice were immunized orally with the recombinant L. acidophilus pMG36e-E0-LA-5. The serum IgG antibody and fecal sIgA antibody responses, expression levels of interleukin (IL)-12 (IL-12) and interferon gamma (IFN-γ) were detected respectively. On the 7th day after the last-immunization, the mice were inoculated with BVDV to evaluate the protective efficiency of the recombinant L. acidophilus pMG36e-E0-LA-5. The results showed that the expressed products protein E0 in the L. acidophilus LA-5 resulted in single band of 27kDa by SDS-PAGE and its strong reactivity with BVDV antibody was confirmed by Western blotting. The IgG and sIgA antibodies responses, IL-12 and IFN-γ expression levels in the vaccinated mice with recombinant L. acidophilus pMG36e-E0-LA-5 were significantly higher than those in the control mice. The protective rate of the vaccinated mice against BVDV increased significantly, and a 90.00% protection rate in virulent challenge was observed. These results indicated that the recombinant L. acidophilus pMG36e-E0-LA-5 strain was successfully constructed and it could effectively improve the immune response in mice and might provide protection against BVDV. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Dietary Lactobacillus acidophilus modulated skin mucus protein profile, immune and appetite genes expression in gold fish (Carassius auratus gibelio).

    PubMed

    Hosseini, Marjan; Kolangi Miandare, Hamed; Shabani, Ali; Hoseinifar, Seyed Hossein; Yarahmadi, Peyman

    2016-12-01

    The objective of the present study was to investigate the effect of dietary Lactobacillus acidophilus on skin mucus protein pattern, immune and appetite related genes expression as well as growth performance in gold fish (Carassius auratus gibelio). Three hundred healthy gold fish (2.5 ± 0.05) juveniles were randomly distributed in 12 glass aquariums (400-L; 25 fish per aquaria) and fed experimental diets contain different levels of L. acidophilus (0, 1.5 × 10 8 , 3 × 10 8 and 6 × 10 8 ) for 8 weeks. SDS-PAGE analysis of skin mucus protein profile at the end of the feeding trial revealed differences in protein profile of probiotic fed fish and control group; even three new bands were observed in L. acidophilus treated groups. Furthermore, fish fed 6 × 10 8  CFU g -1 supplemented diet showed up-regulation of both TNF-1α and TNF-2α gene expression (P < 0.05). Evaluation of appetite related gene expression showed down-regulation of ghrelin in probiotic fed fish compared those of control treatment (P < 0.05). However, administration of different levels of L. acidophilus had no significant effects on growth performance (P > 0.05). These results demonstrated that while no beneficial effects on growth performance, dietary L. acidophilus affects immune and appetite related genes expression as well as skin mucus protein profile. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Probiotic potential and biotherapeutic effects of newly isolated vaginal Lactobacillus acidophilus 36YL strain on cancer cells.

    PubMed

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

    2014-08-01

    Lactobacillus acidophilus is categorized as a probiotic strain because of its beneficial effects in human health and prevention of disease transmission. This study is aimed to characterize the probiotic potential of L. acidophilus 36YL originally isolated from the vagina of healthy and fertile Iranian women. The L. acidophilus 36YL strain was identified using 16S rDNA gene sequencing and characterized by biochemical methodologies, such as antibiotics susceptibility, antimicrobial activity, and acid and bile resistance. The bioactivity of the secretion of this strain on four human cancer cell lines (AGS, HeLa, MCF-7, and HT-29) and one normal cell line (HUVEC) was evaluated by cytotoxicity assay and apoptosis analysis. This newly isolated strain was found to exhibit notable probiotic properties, such as admirable antibiotic susceptibility, good antimicrobial activity, and favorable resistance to acid and bile salt. The results of bioactivity assessment demonstrated acceptable anticancer effects on the four tested cancer cell lines and negligible side effects on the assayed normal cell line. Our findings revealed that the anticancer effect of L. acidophilus 36YL strain secretions depends on the induction of apoptosis in cancer cells. L. acidophilus 36YL strain is considered as a nutraceutical alternative or a topical medication with a potential therapeutic index because of the absence of cytotoxicity to normal cells, but effective toxicity to cancer cell lines. Copyright © 2014 Elsevier Ltd. All rights reserved.

  4. SIGNR3-dependent immune regulation by Lactobacillus acidophilus surface layer protein A in colitis.

    PubMed

    Lightfoot, Yaíma L; Selle, Kurt; Yang, Tao; Goh, Yong Jun; Sahay, Bikash; Zadeh, Mojgan; Owen, Jennifer L; Colliou, Natacha; Li, Eric; Johannssen, Timo; Lepenies, Bernd; Klaenhammer, Todd R; Mohamadzadeh, Mansour

    2015-04-01

    Intestinal immune regulatory signals govern gut homeostasis. Breakdown of such regulatory mechanisms may result in inflammatory bowel disease (IBD). Lactobacillus acidophilus contains unique surface layer proteins (Slps), including SlpA, SlpB, SlpX, and lipoteichoic acid (LTA), which interact with pattern recognition receptors to mobilize immune responses. Here, to elucidate the role of SlpA in protective immune regulation, the NCK2187 strain, which solely expresses SlpA, was generated. NCK2187 and its purified SlpA bind to the C-type lectin SIGNR3 to exert regulatory signals that result in mitigation of colitis, maintenance of healthy gastrointestinal microbiota, and protected gut mucosal barrier function. However, such protection was not observed in Signr3(-/-) mice, suggesting that the SlpA/SIGNR3 interaction plays a key regulatory role in colitis. Our work presents critical insights into SlpA/SIGNR3-induced responses that are integral to the potential development of novel biological therapies for autoinflammatory diseases, including IBD. © 2015 The Authors.

  5. Potential prebiotic properties of cashew apple (Anacardium occidentale L.) agro-industrial byproduct on Lactobacillus species.

    PubMed

    Duarte, Francisca Nayara Dantas; Rodrigues, Jéssica Bezerra; da Costa Lima, Maiara; Lima, Marcos Dos Santos; Pacheco, Maria Teresa Bertoldo; Pintado, Maria Manuela Estevez; de Souza Aquino, Jailane; de Souza, Evandro Leite

    2017-08-01

    The prebiotic effects of a cashew apple (Anacardium occidentale L.) agro-industrial byproduct powder (CAP) on different potentially probiotic Lactobacillus strains, namely Lactobacillus acidophilus LA-05, Lactobacillus casei L-26 and Lactobacillus paracasei L-10, were assessed using in vitro experimental models. Accordingly, the growth of the Lactobacillus strains when cultivated in a broth containing CAP (20 or 30 g L -1 ), glucose (20 g L -1 ) or fructooligosaccharides (FOS) (20 g L -1 ) was monitored over 48 h; the prebiotic activity scores of CAP were determined; and the changes in pH values, production of organic acids and consumption of sugars in growth media were verified. During the 48-h cultivation, similar viable cell counts were observed for the Lactobacillus strains grown in the different media tested. The CAP presented positive prebiotic activity scores toward all the tested Lactobacillus strains, indicating a desirable selective fermentable activity relative to enteric organisms. The cultivation of the Lactobacillus strains in broth containing glucose, FOS or CAP resulted in high viable cell counts, a decreased pH, the production of organic acids and the consumption of sugars over time, revealing intense bacterial metabolic activity. The CAP exerts potential prebiotic effects on different potentially probiotic Lactobacillus strains and should be an added-value ingredient for the food industry. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  6. Probiotic Therapy: A Promising Strategy for the Control of Canine Hookworm

    PubMed Central

    Coêlho, Matheus Diniz Gonçalves; Coêlho, Francine Alves da Silva; de Mancilha, Ismael Maciel

    2013-01-01

    Canis familiaris is a domestic animal which plays an important role as a pet; however, it is very likely to develop diseases characterized by high morbidity and mortality. In this sense, there are some Ancylostomatidae species that can lead to anemia, weight loss, and abdominal pain. Therefore, the treatment with allopathic drugs is essential for eliminating the parasitism. However, the increasing occurrence of resistance to allopathic treatments stimulates the search for new therapeutic inputs, among which the probiotics stand out and are able to positively alter the intestinal microbiota and exert immunomodulatory effect. Therefore, the present study aimed to evaluate the applicability of using species of Lactobacillus (L. acidophilus ATCC 4536, L. plantarum ATCC 8014, and L. delbrueckii UFV H2B20) to control canine ancylostomiasis. A probiotic preparation containing 1 × 106 CFU of each strain was administered to 10 naturally infected animals, on alternate days for 28 days. It was observed that the treatment with the probiotic preparation led to a significant reduction in EPG of feces (88.83%/ P < 0.05), as well as an increase in leukocyte and lymphocyte counts, highlighting the potential use of Lactobacillus species in the control of canine ancylostomiasis. PMID:24386558

  7. Discrimination of the Lactobacillus acidophilus group using sequencing, species-specific PCR and SNaPshot mini-sequencing technology based on the recA gene.

    PubMed

    Huang, Chien-Hsun; Chang, Mu-Tzu; Huang, Mu-Chiou; Wang, Li-Tin; Huang, Lina; Lee, Fwu-Ling

    2012-10-01

    To clearly identify specific species and subspecies of the Lactobacillus acidophilus group using phenotypic and genotypic (16S rDNA sequence analysis) techniques alone is difficult. The aim of this study was to use the recA gene for species discrimination in the L. acidophilus group, as well as to develop a species-specific primer and single nucleotide polymorphism primer based on the recA gene sequence for species and subspecies identification. The average sequence similarity for the recA gene among type strains was 80.0%, and most members of the L. acidophilus group could be clearly distinguished. The species-specific primer was designed according to the recA gene sequencing, which was employed for polymerase chain reaction with the template DNA of Lactobacillus strains. A single 231-bp species-specific band was found only in L. delbrueckii. A SNaPshot mini-sequencing assay using recA as a target gene was also developed. The specificity of the mini-sequencing assay was evaluated using 31 strains of L. delbrueckii species and was able to unambiguously discriminate strains belonging to the subspecies L. delbrueckii subsp. bulgaricus. The phylogenetic relationships of most strains in the L. acidophilus group can be resolved using recA gene sequencing, and a novel method to identify the species and subspecies of the L. delbrueckii and L. delbrueckii subsp. bulgaricus was developed by species-specific polymerase chain reaction combined with SNaPshot mini-sequencing. Copyright © 2012 Society of Chemical Industry.

  8. SIGNR3-dependent immune regulation by Lactobacillus acidophilus surface layer protein A in colitis

    PubMed Central

    Lightfoot, Yaíma L; Selle, Kurt; Yang, Tao; Goh, Yong Jun; Sahay, Bikash; Zadeh, Mojgan; Owen, Jennifer L; Colliou, Natacha; Li, Eric; Johannssen, Timo; Lepenies, Bernd; Klaenhammer, Todd R; Mohamadzadeh, Mansour

    2015-01-01

    Intestinal immune regulatory signals govern gut homeostasis. Breakdown of such regulatory mechanisms may result in inflammatory bowel disease (IBD). Lactobacillus acidophilus contains unique surface layer proteins (Slps), including SlpA, SlpB, SlpX, and lipoteichoic acid (LTA), which interact with pattern recognition receptors to mobilize immune responses. Here, to elucidate the role of SlpA in protective immune regulation, the NCK2187 strain, which solely expresses SlpA, was generated. NCK2187 and its purified SlpA bind to the C-type lectin SIGNR3 to exert regulatory signals that result in mitigation of colitis, maintenance of healthy gastrointestinal microbiota, and protected gut mucosal barrier function. However, such protection was not observed in Signr3−/− mice, suggesting that the SlpA/SIGNR3 interaction plays a key regulatory role in colitis. Our work presents critical insights into SlpA/SIGNR3-induced responses that are integral to the potential development of novel biological therapies for autoinflammatory diseases, including IBD. PMID:25666591

  9. Can Lactobacillus fermentum LF10 and Lactobacillus acidophilus LA02 in a slow-release vaginal product be useful for prevention of recurrent vulvovaginal candidiasis?: A clinical study.

    PubMed

    Murina, Filippo; Graziottin, Alessandra; Vicariotto, Franco; De Seta, Francesco

    2014-01-01

    To assess the effectiveness of the association of 2 specific strains, Lactobacillus fermentum LF10 (DSM 19187) and Lactobacillus acidophilus LA02 (DSM 21717), specifically formulated in slow-release effervescent tablets, in patients with recurrent vulvovaginal candidiasis. The study was a clinical trial of 58 women diagnosed with recurrent VVC (≥4 culture-confirmed episodes in a 12-mo period). All patients were given 200 mg of fluconazole orally as an induction dose for 3 alternate days during the first treatment week. Afterward, the patients were given a new product formulated in slow-release vaginal tablets containing at least 0.4 billion live cells of each of lactobacillus L. fermentum LF10 and L. acidophilus LA02 (first phase of the prophylactic period), on alternate days for 10 consecutive nights. Patients who were still free of symptoms were given 1 vaginal tablet every week for the next 10 weeks (second phase of the prophylactic period). Patients asymptomatic after the total duration of the observation phase (7 mo) were considered as responders. During the second 10-week prophylactic phase, 49 of 57 (86.0%) patients remained free of clinical recurrence, whereas symptomatic VVC occurred in 8 patients (14.0%). During the 7-month follow-up, 42 patients of 49 (85.7%) were symptom free at the end of the protocol, whereas clinical recurrences occurred in 7 women (14.3%). Overall, 42 of 58 women enrolled in the study (72.4%) experienced no clinical recurrence throughout the 7-month observation phase (responders). This study strengthens the evidence supporting the use of specific lactobacilli with well-demonstrated activities associated with the creation and maintenance of a vaginal biofilm that hinders the persistence of an infection caused by Candida.

  10. Lactobacillus acidophilus stimulates the expression of SLC26A3 via a transcriptional mechanism

    PubMed Central

    Raheja, Geetu; Singh, Varsha; Ma, Ke; Boumendjel, Redouane; Borthakur, Alip; Gill, Ravinder K.; Saksena, Seema; Alrefai, Waddah A.; Ramaswamy, Krishnamurthy

    2010-01-01

    Clinical efficacy of probiotics in treating various forms of diarrhea has been clearly established. However, mechanisms underlying antidiarrheal effects of probiotics are not completely defined. Diarrhea is caused either by decreased absorption or increased secretion of electrolytes and solutes in the intestine. In this regard, the electroneutral absorption of two major electrolytes, Na+ and Cl−, occurs mainly through the coupled operation of Na+/H+ exchangers and Cl−/OH− exchangers. Previous studies from our laboratory have shown that Lactobacillus acidophilus (LA) acutely stimulated Cl−/OH− exchange activity via an increase in the surface levels of the apical anion exchanger SLC26A3 (DRA). However, whether probiotics influence SLC26A3 expression and promoter activity has not been examined. The present studies were, therefore, undertaken to investigate the long-term effects of LA on SLC26A3 expression and promoter activity. Treatment of Caco-2 cells with LA for 6–24 h resulted in a significant increase in Cl−/OH− exchange activity. DRA mRNA levels were also significantly elevated in response to LA treatment starting as early as 8 h. Additionally, the promoter activity of DRA was increased by more than twofold following 8 h LA treatment of Caco-2 cells. Similar to the in vitro studies, in vivo studies using mice gavaged with LA also showed significantly increased DRA mRNA (∼4-fold) and protein expression in the colonic regions as assessed by Western blot analysis and immunofluorescence. In conclusion, increase in DRA promoter activity and expression may contribute to the upregulation of intestinal electrolyte absorption and might underlie the potential antidiarrheal effects of LA. PMID:20044511

  11. Astragalus Root and Elderberry Fruit Extracts Enhance the IFN-β Stimulatory Effects of Lactobacillus acidophilus in Murine-Derived Dendritic Cells

    PubMed Central

    Frøkiær, Hanne; Henningsen, Louise; Metzdorff, Stine Broeng; Weiss, Gudrun; Roller, Marc; Flanagan, John; Fromentin, Emilie; Ibarra, Alvin

    2012-01-01

    Many foods and food components boost the immune system, but little data are available regarding the mechanisms by which they do. Bacterial strains have disparate effects in stimulating the immune system. Indendritic cells, the gram-negative bacteria Escherichia coli upregulates proinflammatory cytokines, whereas gram-positive Lactobacillus acidophilus induces a robust interferon (IFN)-β response. The immune-modulating effects of astragalus root and elderberry fruit extracts were examined in bone marrow-derived murine dendritic cells that were stimulated with L. acidophilus or E. coli. IFN-β and other cytokines were measured by ELISA and RT-PCR. Endocytosis of fluorescence-labeled dextran and L. acidophilus in the presence of elderberry fruit or astragalus root extract was evaluated in dendritic cells. Our results show that both extracts enhanced L. acidophilus-induced IFN-β production and slightly decreased the proinflammatory response to E. coli. The enhanced IFN-β production was associated with upregulation of toll-like receptor 3 and to a varying degree, the cytokines IL-12, IL-6, IL-1β and TNF-α. Both extracts increased endocytosis in immature dendritic cells, and only slightly influenced the viability of the cells. In conclusion, astragalus root and elderberry fruit extracts increase the IFN-β inducing activity of L. acidophilus in dendritic cells, suggesting that they may exert antiviral and immune-enhancing activity. PMID:23118903

  12. Characterization of a lactose-responsive promoter of ATP-binding cassette (ABC) transporter gene from Lactobacillus acidophilus 05-172.

    PubMed

    Zeng, Zhu; Zuo, Fanglei; Yu, Rui; Zhang, Bo; Ma, Huiqin; Chen, Shangwu

    2017-09-01

    A novel lactose-responsive promoter of the ATP-binding cassette (ABC) transporter gene Lba1680 of Lactobacillus acidophilus strain 05-172 isolated from a traditionally fermented dairy product koumiss was characterized. In L. acidophilus 05-172, expression of Lba1680 was induced by lactose, with lactose-induced transcription of Lba1680 being 6.1-fold higher than that induced by glucose. This is in contrast to L. acidophilus NCFM, a strain isolated from human feces, in which expression of Lba1680 and Lba1679 is induced by glucose. Both gene expression and enzyme activity assays in L. paracasei transformed with a vector containing the inducible Lba1680 promoter (PLba1680) of strain 05-172 and a heme-dependent catalase gene as reporter confirmed that PLba1680 is specifically induced by lactose. Its regulatory expression could not be repressed by glucose, and was independent of cAMP receptor protein. This lactose-responsive promoter might be used in the expression of functional genes in L. paracasei incorporated into a lactose-rich environment, such as dairy products. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  13. The NAD+-dependent deacetylase, Bifidobacterium longum Sir2 in response to oxidative stress by deacetylating SigHH) and FOXO3a in Bifidobacterium longum and HEK293T cell respectively.

    PubMed

    Guo, Qing; Li, Shiyu; Xie, Yajie; Zhang, Qian; Liu, Mengge; Xu, Zhenrui; Sun, Hanxiao; Yang, Yan

    2017-07-01

    Silent information regulator 2 (Sir2) enzymes which catalyze NAD+-dependent protein/histone deacetylation. The mammalian sirtuin family SIRT1, SIRT2, SIRT3 and SIRT6 can regulate oxidative stress. The probiotics (Bifidobacterium longum(B.longum) and Lactobacillus acidophilus(L. acidophilus)) have Sir2 gene family and have antioxidant activity in human body. it remains unknown whether probiotics Sir2 has a direct role in regulating oxidative stress. To this end, we knockout BL-sir2(sir2 B. longum) and LA-sir2(sir2 L.acidophilus) in low oxygen level. The antioxidant activities of two sir2 deficient strains was decreased, while when reintroduction of BL-sir2 and LA-sir2, the antioxidant activities were recoveried. In order to understand the regulation mechanism of probiotics Sir2 oxidation response. Then, we screened 65 acetylated protein, and found that SigHH ) was a substrate of BL-Sir2. In addition, the acetylation level of σ H decreased with the increase of BL-Sir2 level in B. longum. Thus, BL-Sir2 deacetylated σ H in response to oxidative stress. Next, we transfected BL-Sir2 into H 2 O 2 -induced oxidative damage of 293T cells, BL-Sir2 increased the activity of manganese superoxide dismutase (MnSOD/SOD2) and catalase (CAT) and reduced reactive oxygen species(ROS). Then, we analyzed the differential gene by RNA sequencing and Gene ontology (GO) and found that BL-Sir2 regulated forkhead transcription factor (FOXO3a) mediated antioxidant genes in overexpressed BL-Sir2 HEK293T cells. Our study is the first to link probiotics Sir2 with oxidative stress and uncover the antioxidant mechanism of BL-Sir2 in B. longum itself and human body. Copyright © 2017 Elsevier Inc. All rights reserved.

  14. Influence of osmotic stress on the profile and gene expression of surface layer proteins in Lactobacillus acidophilus ATCC 4356.

    PubMed

    Palomino, María Mercedes; Waehner, Pablo M; Fina Martin, Joaquina; Ojeda, Paula; Malone, Lucía; Sánchez Rivas, Carmen; Prado Acosta, Mariano; Allievi, Mariana C; Ruzal, Sandra M

    2016-10-01

    In this work, we studied the role of surface layer (S-layer) proteins in the adaptation of Lactobacillus acidophilus ATCC 4356 to the osmotic stress generated by high salt. The amounts of the predominant and the auxiliary S-layer proteins SlpA and SlpX were strongly influenced by the growth phase and high-salt conditions (0.6 M NaCl). Changes in gene expression were also observed as the mRNAs of the slpA and slpX genes increased related to the growth phase and presence of high salt. A growth stage-dependent modification on the S-layer protein profile in response to NaCl was observed: while in control conditions, the auxiliary SlpX protein represented less than 10 % of the total S-layer protein, in high-salt conditions, it increased to almost 40 % in the stationary phase. The increase in S-layer protein synthesis in the stress condition could be a consequence of or a way to counteract the fragility of the cell wall, since a decrease in the cell wall thickness and envelope components (peptidoglycan layer and lipoteichoic acid content) was observed in L. acidophilus when compared to a non-S-layer-producing species such as Lactobacillus casei. Also, the stationary phase and growth in high-salt medium resulted in increased release of S-layer proteins to the supernatant medium. Overall, these findings suggest that pre-growth in high-salt conditions would result in an advantage for the probiotic nature of L. acidophilus ATCC 4356 as the increased amount and release of the S-layer might be appropriate for its antimicrobial capacity.

  15. Dual Effects of Cell Free Supernatants from Lactobacillus acidophilus and Lactobacillus rhamnosus GG in Regulation of MMP-9 by Up-Regulating TIMP-1 and Down-Regulating CD147 in PMA- Differentiated THP-1 Cells

    PubMed Central

    Maghsood, Faezeh; Mirshafiey, Abbas; Farahani, Mohadese M.; Modarressi, Mohammad Hossein; Jafari, Parvaneh; Motevaseli, Elahe

    2018-01-01

    Objective Recent studies have reported dysregulated expression of matrix metalloproteinases (MMPs), especially MMP-2, MMP-9, tissue inhibitor of metalloproteinase-1, -2 (TIMP-1, TIMP-2), and extracellular matrix metalloproteinase inducer (EMMPRIN/CD147) in activated macrophages of patients with inflammatory diseases. Therefore, MMP-2, MMP-9, and their regulators may represent a new target for treatment of inflammatory diseases. Probiotics, which are comprised of lactic acid bacteria, have the potential to modulate inflammatory responses. In this experimental study, we investigated the anti-inflammatory effects of cell-free supernatants (CFS) from Lactobacillus acidophilus (L. acidophilus) and L. rhamnosus GG (LGG) in phorbol myristate acetate (PMA)-differentiated THP-1 cells. Materials and Methods In this experimental study, PMA-differentiated THP-1 cells were treated with CFS from L. acidophilus, LGG and uninoculated bacterial growth media (as a control). The expression of MMP-2, MMP-9, TIMP-1, and TIMP-2 mRNAs were determined using real-time quantitative reverse transcription polymerase chain reaction (RT- PCR). The levels of cellular surface expression of CD147 were assessed by flow cytometry, and the gelatinolytic activity of MMP-2 and MMP-9 were determined by zymography. Results Our results showed that CFS from both L. acidophilus and LGG significantly inhibited the gene expression of MMP-9 (P=0.0011 and P=0.0005, respectively), increased the expression of TIMP-1 (P<0.0001), decreased the cell surface expression of CD147 (P=0.0307 and P=0.0054, respectively), and inhibited the gelatinolytic activity of MMP-9 (P=0.0003 and P<0.0001, respectively) in PMA-differentiated THP-1 cells. Although, MMP-2 expression and activity and TIMP-2 expression remained unchanged. Conclusion Our results indicate that CFS from L. acidophilus and LGG possess anti-inflammatory properties and can modulate the inflammatory response. PMID:29105390

  16. Probiotics L. acidophilus and B. clausii Modulate Gut Microbiota in Th1- and Th2-Biased Mice to Ameliorate Salmonella Typhimurium-Induced Diarrhea.

    PubMed

    Pradhan, Biswaranjan; Guha, Dipanjan; Naik, Aman Kumar; Banerjee, Arka; Tambat, Subodh; Chawla, Saurabh; Senapati, Shantibhusan; Aich, Palok

    2018-06-16

    Gut microbiota play important role in maintaining health. Probiotics are believed to augment it further. We aimed at comparing effects of probiotics, Lactobacillus acidophilus (LA) and Bacillus clausii (BC) (a) on the gut microbiota abundance and diversity and (b) their contributions to control intestinal dysbiosis and inflammation in Th1- and Th2-biased mice following Salmonella infection. We report how could gut microbiota and the differential immune bias (Th1 or Th2) of the host regulate host responses when challenged with Salmonella typhimurium in the presence and absence of either of the probiotics. LA was found to be effective in ameliorating the microbial dysbiosis and inflammation caused by Salmonella infection, in Th1 (C57BL/6) and Th2 (BALB/c)-biased mouse. BC was able to ameliorate Salmonella-induced dysbiosis and inflammation in Th2 but not in Th1-biased mouse. These results may support probiotics LA as a treatment option in the case of Salmonella infection.

  17. Cloning, expression and characterization of a mucin-binding GAPDH from Lactobacillus acidophilus.

    PubMed

    Patel, Dhaval K; Shah, Kunal R; Pappachan, Anju; Gupta, Sarita; Singh, Desh Deepak

    2016-10-01

    Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a ubiquitous enzyme involved in glycolysis. It is also referred to as a moonlighting protein as it has many diverse functions like regulation of apoptosis, iron homeostasis, cell-matrix interactions, adherence to human colon etc. apart from its principal role in glycolysis. Lactobacilli are lactic acid bacteria which colonize the human gut and confer various health benefits to humans. In the present study, we have cloned, expressed and purified the GAPDH from Lactobacillus acidophilus to get a recombinant product (r-LaGAPDH) and characterized it. Size exclusion chromatography shows that r-LaGAPDH exists as a tetramer in solution and have a mucin binding and hemagglutination activity indicating carbohydrate like binding adhesion mechanism. Fluorescence spectroscopy studies showed an interaction of r-LaGAPDH with mannose, galactose, N-acetylgalactosamine and N-acetylglucosamine with a Kd of 3.6±0.7×10(-3)M, 4.34±0.09×10(-3)M, 4±0.87×10(-3)M and 3.7±0.28×10(-3)M respectively. We hope that this preliminary data will generate more interest in further elucidation of the roles of GAPDH in the adhesion processes of the bacteria. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Enzymology and structure of the GH13_31 glucan 1,6-α-glucosidase that confers isomaltooligosaccharide utilization in the probiotic Lactobacillus acidophilus NCFM.

    PubMed

    Møller, Marie S; Fredslund, Folmer; Majumder, Avishek; Nakai, Hiroyuki; Poulsen, Jens-Christian N; Lo Leggio, Leila; Svensson, Birte; Abou Hachem, Maher

    2012-08-01

    Isomaltooligosaccharides (IMO) have been suggested as promising prebiotics that stimulate the growth of probiotic bacteria. Genomes of probiotic lactobacilli from the acidophilus group, as represented by Lactobacillus acidophilus NCFM, encode α-1,6 glucosidases of the family GH13_31 (glycoside hydrolase family 13 subfamily 31) that confer degradation of IMO. These genes reside frequently within maltooligosaccharide utilization operons, which include an ATP-binding cassette transporter and α-glucan active enzymes, e.g., maltogenic amylases and maltose phosphorylases, and they also occur separated from any carbohydrate transport or catabolism genes on the genomes of some acidophilus complex members, as in L. acidophilus NCFM. Besides the isolated locus encoding a GH13_31 enzyme, the ABC transporter and another GH13 in the maltooligosaccharide operon were induced in response to IMO or maltotetraose, as determined by reverse transcription-PCR (RT-PCR) transcriptional analysis, suggesting coregulation of α-1,6- and α-1,4-glucooligosaccharide utilization loci in L. acidophilus NCFM. The L. acidophilus NCFM GH13_31 (LaGH13_31) was produced recombinantly and shown to be a glucan 1,6-α-glucosidase active on IMO and dextran and product-inhibited by glucose. The catalytic efficiency of LaGH13_31 on dextran and the dextran/panose (trisaccharide) efficiency ratio were the highest reported for this class of enzymes, suggesting higher affinity at distal substrate binding sites. The crystal structure of LaGH13_31 was determined to a resolution of 2.05 Å and revealed additional substrate contacts at the +2 subsite in LaGH13_31 compared to the GH13_31 from Streptococcus mutans (SmGH13_31), providing a possible structural rationale to the relatively high affinity for dextran. A comprehensive phylogenetic and activity motif analysis mapped IMO utilization enzymes from gut microbiota to rationalize preferential utilization of IMO by gut residents.

  19. Enzymology and Structure of the GH13_31 Glucan 1,6-α-Glucosidase That Confers Isomaltooligosaccharide Utilization in the Probiotic Lactobacillus acidophilus NCFM

    PubMed Central

    Møller, Marie S.; Fredslund, Folmer; Majumder, Avishek; Nakai, Hiroyuki; Poulsen, Jens-Christian N.; Lo Leggio, Leila; Svensson, Birte

    2012-01-01

    Isomaltooligosaccharides (IMO) have been suggested as promising prebiotics that stimulate the growth of probiotic bacteria. Genomes of probiotic lactobacilli from the acidophilus group, as represented by Lactobacillus acidophilus NCFM, encode α-1,6 glucosidases of the family GH13_31 (glycoside hydrolase family 13 subfamily 31) that confer degradation of IMO. These genes reside frequently within maltooligosaccharide utilization operons, which include an ATP-binding cassette transporter and α-glucan active enzymes, e.g., maltogenic amylases and maltose phosphorylases, and they also occur separated from any carbohydrate transport or catabolism genes on the genomes of some acidophilus complex members, as in L. acidophilus NCFM. Besides the isolated locus encoding a GH13_31 enzyme, the ABC transporter and another GH13 in the maltooligosaccharide operon were induced in response to IMO or maltotetraose, as determined by reverse transcription-PCR (RT-PCR) transcriptional analysis, suggesting coregulation of α-1,6- and α-1,4-glucooligosaccharide utilization loci in L. acidophilus NCFM. The L. acidophilus NCFM GH13_31 (LaGH13_31) was produced recombinantly and shown to be a glucan 1,6-α-glucosidase active on IMO and dextran and product-inhibited by glucose. The catalytic efficiency of LaGH13_31 on dextran and the dextran/panose (trisaccharide) efficiency ratio were the highest reported for this class of enzymes, suggesting higher affinity at distal substrate binding sites. The crystal structure of LaGH13_31 was determined to a resolution of 2.05 Å and revealed additional substrate contacts at the +2 subsite in LaGH13_31 compared to the GH13_31 from Streptococcus mutans (SmGH13_31), providing a possible structural rationale to the relatively high affinity for dextran. A comprehensive phylogenetic and activity motif analysis mapped IMO utilization enzymes from gut microbiota to rationalize preferential utilization of IMO by gut residents. PMID:22685275

  20. Fermentation optimization of goat milk with Lactobacillus acidophilus and Bifidobacterium bifidum by Box-Behnken design.

    PubMed

    Shu, Guowei; Bao, Chunju; Chen, He; Wang, Changfeng; Yang, Hui

    2016-01-01

    Goat milk is only limited to the processing of goat milk powder and liquid milk, the products are mainly about milk powder and a few of them are made as milk tablet. Therefore, the study of probiotic goat milk will have great significance in the full use of goats and the development of the goat milk industry in China. The effect of fermentation temperature (35°C, 37°C, 39°C), strain ratio (1:1:1, 2:1:1, 3:1:1) and inoculum size (4%, 5%, 6%) on viable counts of L. acidophilus and B. bifidum, total bacteria and sensory value during fermentation process of L. acidophilus and B. bifidum goat yogurt (AB-goat yogurt) was investigated. The optimum fermentation conditions for AB-goat yogurt were: fermentation temperature 38°C, the strain ratio 2:1:1, inoculum size 6%. Under the optimum conditions, the viable counts of B. bifidum, L. acidophilus, total bacteria and sensory value reached (4.30 ±0.11)×107  cfu/mL, (1.39 ±0.09)×108  cfu/mL, (1.82±0.06)×109  cfu/mL and 7.90 ±0.14, respectively. The fermentation temperature, the strain ratio and inoculum size had a significant effect on the fermentation of AB-goat yogurt and these results are beneficial for developing AB-goat yogurt.

  1. S-layer proteins from Lactobacillus sp. inhibit bacterial infection by blockage of DC-SIGN cell receptor.

    PubMed

    Prado Acosta, Mariano; Ruzal, Sandra M; Cordo, Sandra M

    2016-11-01

    Many species of Lactobacillus sp. possess Surface(s) layer proteins in their envelope. Among other important characteristics S-layer from Lactobacillus acidophilus binds to the cellular receptor DC-SIGN (Dendritic Cell-Specific Intercellular adhesion molecule-3-Grabbing Non-integrin; CD209), which is involved in adhesion and infection of several families of bacteria. In this report we investigate the activity of new S-layer proteins from the Lactobacillus family (Lactobacillus acidophilus, Lactobacillus brevis, Lactobacillus helveticus and Lactobacillus kefiri) over the infection of representative microorganisms important to human health. After the treatment of DC-SIGN expressing cells with these proteins, we were able to diminish bacterial infection by up to 79% in both gram negative and mycobacterial models. We discovered that pre-treatment of the bacteria with S-layers from Lactobacillus acidophilus and Lactobacillus brevis reduced bacteria viability but also prevent infection by the pathogenic bacteria. We also proved the importance of the glycosylation of the S-layer from Lactobacillus kefiri in the binding to the receptor and thus inhibition of infection. This novel characteristic of the S-layers proteins may contribute to the already reported pathogen exclusion activity for these Lactobacillus probiotic strains; and might be also considered as a novel enzymatic antimicrobial agents to inhibit bacterial infection and entry to host cells. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. Inhibition of H9N2 Virus Invasion into Dendritic Cells by the S-Layer Protein from L. acidophilus ATCC 4356

    PubMed Central

    Gao, Xue; Huang, Lulu; Zhu, Liqi; Mou, Chunxiao; Hou, Qihang; Yu, Qinghua

    2016-01-01

    Probiotics are essential for the prevention of virus invasion and the maintenance of the immune balance. However, the mechanism of competition between probiotics and virus are unknown. The objectives of this study were to isolate the surface layer (S-layer) protein from L. acidophilus ATCC 4356 as a new antiviral material, to evaluate the stimulatory effects of the S-layer protein on mouse dendritic cells (DCs) and to verify its ability to inhibit the invasion of H9N2 avian influenza virus (AIV) in DCs. We found that the S-layer protein induced DCs activation and up-regulated the IL-10 secretion. The invasion and replication of the H9N2 virus in mouse DCs was successfully demonstrated. However, the invasion of H9N2 virus into DCs could be inhibited by treatment with the S-layer protein prior to infection, which was verified by the reduced hemagglutinin (HA) and neuraminidase (NA) mRNA expression, and nucleoprotein (NP) protein expression in the DCs. Furthermore, treatment with the S-layer protein increases the Mx1, Isg15, and Ddx58 mRNA expressions, and remits the inflammatory process to inhibit H9N2 AIV infection. In conclusion, the S-layer protein stimulates the activation of mouse DCs, inhibits H9N2 virus invasion of DCs, and stimulates the IFN-I signaling pathway. Thus, the S-layer protein from Lactobacillus is a promising biological antiviral material for AIV prevention. PMID:27826541

  3. Effects of encapsulated Lactobacillus acidophilus along with pasteurized longan juice on the colon microbiota residing in a dynamic simulator of the human intestinal microbial ecosystem.

    PubMed

    Chaikham, Pittaya; Apichartsrangkoon, Arunee

    2014-01-01

    The effect of encapsulated Lactobacillus acidophilus LA5 along with pasteurized longan juice on the colon microbiota was investigated by applying a dynamic model of the human gastrointestinal tract. Encapsulated L. acidophilus LA5 in pasteurized longan juice or sole encapsulated L. acidophilus LA5 exhibited the efficiency of colonizing the colon and enabling the growth of colon lactobacilli as well as beneficial bifidobacteria but inhibited the growth of fecal coliforms and clostridia. Moreover, these treatments gave rise to a significant increase of lactic acid and short-chain fatty acids such as acetate, propionate, and butyrate. Although acetate displayed the highest quantity, it was likely that after incorporating encapsulated L. acidophilus LA5 plus pasteurized longan juice, quantity of butyrate exceed propionate, and acetate in comparison with their controls. Denaturant gradient gel electrophoresis patterns confirmed that various treatments affected the alteration of microbial community within the simulator of the human intestinal microbial ecosystem.

  4. 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.

  5. Some probiotic and antibacterial properties of Lactobacillus acidophilus cultured from dahi a native milk product.

    PubMed

    Mahmood, Talat; Masud, Tariq; Sohail, Asma

    2014-08-01

    In this study, different strains of Lactobacillus acidophilus from dahi were analyzed for certain probiotic and antibacterial properties. Initially, these strains were confirmed by the amplification of 16S rRNA regions and then screened for antibacterial activities against food borne pathogens. The phenotypic relationship between apparent antibacterial activity and cell wall proteins were established by cluster analysis. It was observed that those strains, which have prominent bands having size 22-25 kDa possess antibacterial activity. On the basis of wide spectrum of killing pattern, a strain LA06FT was further characterized that showed no change in its behavior when subjected to the antibiotic protected environment and grow well in acid-bile conditions. The bacteriocin produced by this strain has specific antibacterial activity of 5369.13 AU mg(-1). It remained stable at 60-90 °C and pH range of 4.5-6.5 while proteolytic enzymes inactivate the bacteriocin that confirm its proteinic nature having molecular weight of ≤8.5 kDa.

  6. Influence of the addition of Lactobacillus acidophilus La-05, Bifidobacterium animalis subsp. lactis Bb-12 and inulin on the technological, physicochemical, microbiological and sensory features of creamy goat cheese.

    PubMed

    Barbosa, Ilsa C; Oliveira, Maria E G; Madruga, Marta S; Gullón, Beatriz; Pacheco, Maria T B; Gomes, Ana M P; Batista, Ana S M; Pintado, Maria M E; Souza, Evandro L; Queiroga, Rita C R E

    2016-10-12

    The effects of the addition of Lactobacillus acidophilus LA-05, Bifidobacterium animalis subsp. lactis BB-12 and inulin on the quality characteristics of creamy goat cheese during refrigerated storage were evaluated. The manufactured cheeses included the addition of starter culture (Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris - R-704) (CC); starter culture, L. acidophilus LA-05 and inulin (CLA); starter culture, B. lactis BB-12 and inulin (CBB); or starter culture, L. acidophilus LA-05, B. lactis BB-12 and inulin (CLB). In the synbiotic cheeses (CLA, CBB and CLB), the counts of L. acidophilus LA-05 and B. lactis BB-12 were greater than 6log CFU g -1 , the amount of inulin was greater than 6 g per 100 g, and the firmness was reduced. The cheeses evaluated had high brightness values (L*), with a predominance of yellow (b*). CC had higher contents of proteins, lipids and minerals compared to the other cheeses. There was a decrease in the amount of short-chain fatty acids (SCFAs) and an increase of medium-chain (MCFAs) and long-chain fatty acids (LCFAs) in the synbiotic cheeses compared to CC. The amount of conjugated linoleic acid increased in CLA, CBB and CLB. The highest depth of proteolysis and the greatest changes in the release of free amino acids were found in CLB. The addition of inulin and probiotics, alone or in co-culture, did not affect the cheese acceptance. Inulin and probiotics can be used together for the production of creamy goat cheese without negatively affecting the general quality characteristics of the product, and to add value because of its synbiotic potential.

  7. Lactobacillus acidophilus INMIA 9602 Er-2 strain 317/402 probiotic regulates growth of commensal Escherichia coli in gut microbiota of familial Mediterranean fever disease subjects.

    PubMed

    Pepoyan, A Z; Balayan, M H; Manvelyan, A M; Mamikonyan, V; Isajanyan, M; Tsaturyan, V V; Kamiya, S; Netrebov, V; Chikindas, M L

    2017-04-01

    Previously, we reported a positive effect the probiotic formulation, Lactobacillus acidophilus INMIA 9602 Er-2 strain 317/402 (Narine strain), had on the blood characteristics of patients with familial Mediterranean fever disease (FMF). The aim of this investigation was to evaluate the effect of the Narine probiotic on growth characteristics in the predominant commensal Escherichia coli isolates from the gut microbiota in FMF-positive study participants. Bacterial growth of 192 prevalent commensal E. coli isolates found in the volunteer participants' guts was evaluated using Verhulst's logistic function. This study showed that the duration of the preparatory growth phase for the E. coli isolates collected from FMF-positive volunteers was significantly shorter, whereas the duration of the logarithmic growth phase was significantly longer (P < 0·03) than that of the isolates collected from healthy participants. The Narine probiotic formulation caused a significant extension (P < 0·001) of the preparatory growth phase in the commensal E. coli isolated from FMF subjects a month after the Narine probiotic administration was terminated. The data suggest that the mathematical model characterizes the growth of commensal E. coli isolates from FMF-positive participants and it can be useful in a decision-making process on the practical use of probiotics during FMF. This is the first study to demonstrate the effects of Narine, containing the probiotic Lactobacillus acidophilus, on the growth of gut commensal Escherichia coli from study participants with familial Mediterranean fever disease (FMF). Verhulst's logistic function was demonstrated to act as a possible tool for the evaluation and quantification of effects produced by the probiotic formulation in FMF participants. © 2017 The Society for Applied Microbiology.

  8. Antimicrobial activity of lactobacillus strains against uropathogens.

    PubMed

    Shim, Yoon Hee; Lee, Seung Joo; Lee, Jung Won

    2016-10-01

    The use of lactobacillus probiotics has been proposed as an alternative to prophylactic antibiotics for preventing urinary tract infection (UTI) in the era of antibiotic resistance. In this study, the antimicrobial activity of lactobacillus strains against uropathogens, was evaluated and compared with that of antibiotics. To evaluate inhibitory activities of lactobacilli against uropathogens, six lactobacillus strains (L. gasseri, L. rhamnosus, L. acidophilus, L. plantarum, L. paracasei, L. acidophilus) and four representative uropathogens of infantile UTI (extended-spectrum beta-lactamase [ESBL](-) Escherichia coli, ESBL(+) E. coli, Proteus vulgaris, Enterococcus fecalis) were selected. Lactobacillus strain in vitro inhibition of each uropathogen was evaluated on MRS agar well diffusion assay and compared with that of commercial antibiotic discs. Average inhibitory zone for each of the six lactobacillus strains against the four uropathogens showed slightly different but consistent inhibition (inhibitory zone diameter, 10.5-20.0 mm). This was different to that of the antibiotic discs, which had a wider range of inhibition (inhibitory zone diameter, <6.0-27.5 mm) depending on the uropathogen resistance pattern. The inhibitory zone of the six lactobacillus strains was between that of sensitive and resistant antibiotics (P < 0.05). Lactobacillus strains had similar moderate antimicrobial activities against uropathogens. Further research is needed to ascertain the strains with the best probiotic potential. © 2016 Japan Pediatric Society.

  9. Transcriptional and functional analysis of galactooligosaccharide uptake by lacS in Lactobacillus acidophilus

    PubMed Central

    Andersen, Joakim M.; Barrangou, Rodolphe; Abou Hachem, Maher; Lahtinen, Sampo; Goh, Yong Jun; Svensson, Birte; Klaenhammer, Todd R.

    2011-01-01

    Probiotic microbes rely on their ability to survive in the gastrointestinal tract, adhere to mucosal surfaces, and metabolize available energy sources from dietary compounds, including prebiotics. Genome sequencing projects have proposed models for understanding prebiotic catabolism, but mechanisms remain to be elucidated for many prebiotic substrates. Although β-galactooligosaccharides (GOS) are documented prebiotic compounds, little is known about their utilization by lactobacilli. This study aimed to identify genetic loci in Lactobacillus acidophilus NCFM responsible for the transport and catabolism of GOS. Whole-genome oligonucleotide microarrays were used to survey the differential global transcriptome during logarithmic growth of L. acidophilus NCFM using GOS or glucose as a sole source of carbohydrate. Within the 16.6-kbp gal-lac gene cluster, lacS, a galactoside-pentose-hexuronide permease-encoding gene, was up-regulated 5.1-fold in the presence of GOS. In addition, two β-galactosidases, LacA and LacLM, and enzymes in the Leloir pathway were also encoded by genes within this locus and up-regulated by GOS stimulation. Generation of a lacS-deficient mutant enabled phenotypic confirmation of the functional LacS permease not only for the utilization of lactose and GOS but also lactitol, suggesting a prominent role of LacS in the metabolism of a broad range of prebiotic β-galactosides, known to selectively modulate the beneficial gut microbiota. PMID:22006318

  10. Antiadherent and antibacterial properties of stainless steel and NiTi orthodontic wires coated with silver against Lactobacillus acidophilus--an in vitro study.

    PubMed

    Mhaske, Arun Rameshwar; Shetty, Pradeep Chandra; Bhat, N Sham; Ramachandra, C S; Laxmikanth, S M; Nagarahalli, Kiran; Tekale, Pawankumar Dnyandeo

    2015-01-01

    The purpose of the study is to assess the antiadherent and antibacterial properties of surface-modified stainless steel and NiTi orthodontic wires with silver against Lactobacillus acidophilus. This study was done on 80 specimens of stainless steel and NiTi orthodontic wires. The specimens were divided into eight test groups. Each group consisted of 10 specimens. Groups containing uncoated wires acted as a control group for their respective experimental group containing coated wires. Surface modification of wires was carried out by the thermal vacuum evaporation method with silver. Wires were then subjected to microbiological tests for assessment of the antiadherent and antibacterial properties of silver coating against L. acidophilus. Mann-Whitney U test was used to analyze the colony-forming units (CFUs) in control and test groups; and Student's t test (two-tailed, dependent) was used to find the significance of study parameters on a continuous scale within each group. Orthodontic wires coated with silver showed an antiadherent effect against L. acidophilus compared with uncoated wires. Uncoated stainless steel and NiTi wires respectively showed 35.4 and 20.5 % increase in weight which was statistically significant (P < 0.001), whereas surface-modified wires showed only 4.08 and 4.4 % increase in weight (statistically insignificant P > 0.001). The groups containing surface-modified wires showed statistically significant decrease in the survival rate of L. acidophilus expressed as CFU and as log of colony count when compared to groups containing uncoated wires. It was 836.60 ± 48.97 CFU in the case of uncoated stainless steel whereas it was 220.90 ± 30.73 CFU for silver-modified stainless steel, 748.90 ± 35.64 CFU for uncoated NiTi, and 203.20 ± 41.94 CFU for surface-modified NiTi. Surface modification of orthodontic wires with silver can be used to prevent the accumulation of dental plaque and the development of dental caries

  11. Lactobacillus acidophilus CP23 with weak immunomodulatory activity lacks anchoring structure for surface layer protein.

    PubMed

    Yanagihara, Sae; Kato, Shinji; Ashida, Nobuhisa; Yamamoto, Naoyuki

    2015-05-01

    To determine the reason for the low levels of Surface layer protein A (SlpA) on CP23 cells, which might play a crucial role in the immunomodulatory effect of Lactobacillus acidophilus, the DNA sequence of the slpA gene of CP23 and L-92 strains, including the upstream region, were analyzed. Unexpectedly, there was no significant difference in the predicted amino acid sequence of the C-terminus needed for cell anchoring, and only an additional Ala-Val-Ala sequence inserted in the N-terminal region of the mature CP23 protein. Therefore, anchoring of SlpA on the cell wall of CP23 and L-92 was evaluated by a reconstitution assay, which showed that SlpA released by LiCl treatment from both CP23 and L-92 was successfully anchored on LiCl-treated L-92 cells, but not on LiCl-treated CP23 cells. Moreover, quantitative analysis of SlpA protein in the culture medium of CP23 and L-92 by ELISA revealed higher levels of SlpA secretion in CP23 cells than in L-92 cells. Collectively, these results suggest that the lower levels of SlpA on the surface of CP23 cells might be caused by less cell wall capacity for SlpA anchoring, leading to an accumulation of SlpA in the culture medium of CP23 cells. The present study supports the importance of cell surface structure of L. acidophilus L-92 for SlpA anchoring on the cell surface needed for immunomodulatory effect. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  12. [Influence of corynebacteria metabolites on antagonistic activity of H2O2 producing lactobacilli].

    PubMed

    Bukharin, O V; Sgibnev, A V

    2012-01-01

    Study combined influence of Corynebacterium genus bacteria metabolites and H2O2 producing lactobacilli on survival rate of Staphylococcus aureus, Escherichia coli and Lactobacillus acidophilus. The ability to inhibit catalase of the test strains used and to reduce bactericidal effect of hydroxyl radical were determined in corynebacteria. H2O2 containing metabolites were obtained by cultivating lactobacilli in mineral medium, the amount of H2O2 was determined by oxidation of TMB by peroxidase. Bactericidal effect of lactobacilli metabolites for test strains treated by corynebacteria metabolites was evaluated by seeding results. Results. Inhibitio by corynebacteria metabolites of S. aureus catalase activity by 30-40% and E. coli catalase activ ity by 40-70% was shown. A reduction of bactericidal effect of hydroxyl radicals by corynebacteria metabolites by 30-35% for S. aureus, 38-42% for E. coli and 70-73% for L. acidophilus was noted. The enchantment of bactericidal effect of lactobacilli after treatment of the test strain by corynebacteria metabolites against S. aureus and E. coli manifested by reduction of the numbe of viable cells by 2-3 lg CFU. For L. acidophilus the bactericidal effect oflactobacilli metabolite in the same conditions reduced, and that led to the increase ofviability by 2-4 lg PFU. A conclusion on the possibility of regulation by associative bacteria the manifestations of antagonistic activity of H2O2 producing dominant microorganisms is made based on the data obtained.

  13. Selection of the Strain Lactobacillus acidophilus ATCC 43121 and Its Application to Brewers' Spent Grain Conversion into Lactic Acid

    PubMed Central

    Liguori, Rossana; Soccol, Carlos Ricardo; Vandenberghe, Luciana Porto de Souza; Woiciechowski, Adenise Lorenci; Ionata, Elena; Marcolongo, Loredana; Faraco, Vincenza

    2015-01-01

    Six Lactobacillus strains were analyzed to select a bacterium for conversion of brewers' spent grain (BSG) into lactic acid. Among the investigated strains, L. acidophilus ATCC 43121 showed the highest yield of lactic acid production (16.1 g/L after 48 hours) when grown in a synthetic medium. It was then analyzed for its ability to grow on the hydrolysates obtained from BSG after acid-alkaline (AAT) or aqueous ammonia soaking (AAS) pretreatment. The lactic acid production by L. acidophilus ATCC 43121 through fermentation of the hydrolysate from AAS treated BSG was 96% higher than that from the AAT treated one, although similar yields of lactic acid per consumed glucose were achieved due to a higher (46%) glucose consumption by L. acidophilus ATCC 43121 in the AAS BSG hydrolysate. It is worth noting that adding yeast extract to the BSG hydrolysates increased both the yield of lactic acid per substrate consumed and the volumetric productivity. The best results were obtained by fermentation of AAS BSG hydrolysate supplemented by yeast extract, in which the strain produced 22.16 g/L of lactic acid (yield of 0.61 g/g), 27% higher than the value (17.49 g/L) obtained in the absence of a nitrogen source. PMID:26640784

  14. Impairment of Swimming Motility by Antidiarrheic Lactobacillus acidophilus Strain LB Retards Internalization of Salmonella enterica Serovar Typhimurium within Human Enterocyte-Like Cells▿

    PubMed Central

    Liévin-Le Moal, Vanessa; Amsellem, Raymonde; Servin, Alain L.

    2011-01-01

    We report that both culture and the cell-free culture supernatant (CFCS) of Lactobacillus acidophilus strain LB (Lactéol Boucard) have the ability (i) to delay the appearance of Salmonella enterica serovar Typhimurium strain SL1344-induced mobilization of F-actin and, subsequently, (ii) to retard cell entry by S. Typhimurium SL1344. Time-lapse imaging and Western immunoblotting showed that S. Typhimurium SL1344 swimming motility, as represented by cell tracks of various types, was rapidly but temporarily blocked without affecting the expression of FliC flagellar propeller protein. We show that the product(s) secreted by L. acidophilus LB that supports the inhibitory activity is heat stable and of low molecular weight. The product(s) caused rapid depolarization of the S. Typhimurium SL1344 cytoplasmic membrane without affecting bacterial viability. We identified inhibition of swimming motility as a newly discovered mechanism by which the secreted product(s) of L. acidophilus strain LB retards the internalization of the diarrhea-associated pathogen S. enterica serovar Typhimurium within cultured human enterocyte-like cells. PMID:21825295

  15. Regulation of the Lactobacillus Strains on HMGCoA Reductase Gene Transcription in Human HepG2 Cells via Nuclear Factor-κB.

    PubMed

    Chen, Kun; Li, Shaocong; Chen, Fang; Li, Jun; Luo, Xuegang

    2016-02-01

    Lactic acid bacteria have been identified to be effective in reducing cholesterol levels. Most of the mechanistic studies were focused on the bile salt deconjugation ability of bile salt hydrolase in lactic acid bacteria. However, the mechanism by which Lactobacillus decreases cholesterol levels has not been thoroughly studied in intact primate cells. 3-Hydroxy-3- methyl-glutaryl-coenzyme A reductase (HMGCR) is the vital enzyme in cholesterol synthesis. To confirm the effect of probiotic Lactobacillus strains on HMGCR level, in the present study, human hepatoma HepG2 cells were treated with Lactobacillus strains, and then the HMGCR level was illustrated by luciferase reporter assay and RT-PCR. The results showed that the level of HMGCR was suppressed after being treated with the live Lactobacillus strains. These works might set a foundation for the following study of the antihyperlipidemic effects of L. acidophilus, and contribute to the development of functional foods or drugs that benefit patients suffering from hyperlipidemia diseases.

  16. Application of Lactobacillus acidophilus (LA 5) strain in fruit-based ice cream

    PubMed Central

    Senanayake, Suraji A; Fernando, Sirimali; Bamunuarachchi, Arthur; Arsekularatne, Mariam

    2013-01-01

    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 × 107 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

  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.

  18. Lactobacillus Species Identification, H2O2 Production, and Antibiotic Resistance and Correlation with Human Clinical Status

    PubMed Central

    Felten, Annie; Barreau, Claude; Bizet, Chantal; Lagrange, Philippe Henri; Philippon, Alain

    1999-01-01

    Lactobacilli recovered from the blood, cerebrospinal fluid, respiratory tract, and gut of 20 hospitalized immunocompromised septic patients were analyzed. Biochemical carbohydrate fermentation and total soluble cell protein profiles were used to identify the species. Hydrogen peroxide production was measured. Susceptibility to 19 antibiotics was tested by a diffusion method, and the MICs of benzylpenicillin, amoxicillin, imipenem, erythromycin, vancomycin, gentamicin, and levofloxacin were determined. A small number of species produced H2O2, and antibiotic susceptibilities were species related. Eighteen (90%) of the isolates were L. rhamnosus, one was L. paracasei subsp. paracasei, and one was L. crispatus. L. rhamnosus, L. paracasei subsp. paracasei isolates, and the type strains were neither H2O2 producers nor vancomycin susceptible (MICs, ≥256 μg/ml). L. crispatus, as well as most of the type strains of lactobacilli which belong to the L. acidophilus group, was an H2O2 producer and vancomycin susceptible (MICs, <4 μg/ml). PMID:9986841

  19. Prevalence of Escherichia coli O157:H7 and performance by beef feedlot cattle given Lactobacillus direct-fed microbials.

    PubMed

    Brashears, M M; Galyean, M L; Loneragan, G H; Mann, J E; Killinger-Mann, K

    2003-05-01

    Fecal shedding of Escherichia coli O157:H7, the prevalence of Escherichia coli O157:H7 in pens and on carcasses and hides, and cattle performance as a result of daily dietary supplementation with Lactobacillus-based direct-fed microbials (DFMs) were evaluated in a feeding trial involving 180 beef steers. Steers were evaluated for shedding of E. coli O157:H7 by an immunomagnetic separation technique on arrival at the feedlot, just before treatment with the DFMs, and every 14 days thereafter until slaughter. Composite pen fecal samples were collected every 14 days (alternating weeks with animal testing), and prevalence on hides and carcasses at slaughter was also evaluated. Feedlot performance (body weight gain and feed intake) was measured for the period during which the DFMs were fed. Gain efficiency was calculated as the ratio of weight gain to feed intake. Lactobacillus acidophilus NPC 747 decreased (P < 0.01) the shedding of E. coli O157:H7 in the feces of individual cattle during the feeding period. E. coli O157:H7 was approximately twice as likely to be detected in control animal samples as in samples from animals receiving L. acidophilus NPC 747. In addition, DFM supplementation decreased (P < 0.05) the number of E. coli O157:H7-positive hide samples at harvest and the number of pens testing positive for the pathogen. Body weight gains (on a live or carcass basis) and feed intakes during the DFM supplementation period did not differ among treatments. Gain efficiencies on a live-weight basis did not differ among treatments, but carcass-based gain/feed ratios tended (P < 0.06) to be better for animals receiving the two DFM treatments than for control animals. The results of this study suggest that the feeding of a Lactobacillus-based DFM to cattle will decrease, but not eliminate, fecal shedding of E. coli O157:H7, as well as contamination on hides, without detrimental effects on performance.

  20. A novel unsaturated fatty acid hydratase toward C16 to C22 fatty acids from Lactobacillus acidophilus

    PubMed Central

    Hirata, Akiko; Kishino, Shigenobu; Park, Si-Bum; Takeuchi, Michiki; Kitamura, Nahoko; Ogawa, Jun

    2015-01-01

    Hydroxy FAs, one of the gut microbial metabolites of PUFAs, have attracted much attention because of their various bioactivities. The purpose of this study was to identify lactic acid bacteria with the ability to convert linoleic acid (LA) to hydroxy FAs. A screening process revealed that a gut bacterium, Lactobacillus acidophilus NTV001, converts LA mainly into 13-hydroxy-cis-9-octadecenoic acid and resulted in the identification of the hydratase responsible, fatty acid hydratase 1 (FA-HY1). Recombinant FA-HY1 was purified, and its enzymatic characteristics were investigated. FA-HY1 could convert not only C18 PUFAs but also C20 and C22 PUFAs. C18 PUFAs with a cis carbon-carbon double bond at the Δ12 position were converted into the corresponding 13-hydroxy FAs. Arachidonic acid and DHA were converted into the corresponding 15-hydroxy FA and 14-hydroxy FA, respectively. To the best of our knowledge, this is the first report of a bacterial FA hydratase that can convert C20 and C22 PUFAs into the corresponding hydroxy FAs. These novel hydroxy FAs produced by using FA-HY1 should contribute to elucidating the bioactivities of hydroxy FAs. PMID:25966711

  1. 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

  2. Evaluation of phytate-degrading Lactobacillus culture administration to broiler chickens.

    PubMed

    Askelson, Tyler E; Campasino, Ashley; Lee, Jason T; Duong, Tri

    2014-02-01

    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.

  3. [Diversity of Lactobacillus in vagina of vulvovaginal candidiasis].

    PubMed

    2015-04-07

    To investigate the Lactobacillus species in the vaginas of vulvovaginal candidiasis and to assess the prevalence of each Lactobacillus species in vulvovaginal candidiasis. 154 vaginal samples were analyzed, 92 of which were from fertile healthy women, and 62 of which were from women with vulvovaginal candidiasis; and species-specific PCR showed the prevalence of each Lactobacillus species Species-specific PCR was used to investigate the prevalence of each Lactobacillus species in healthy Chinese women and the women with vulvovaginal candidiasis. In women with vulvovaginal candidiasis: L. iners (6.5%), L. cripatus (79.0%), L. gasseri (37.1%), L. jensenii (74.2%), L. acidophilus (16.1%), L. brevis (19.4%), L. plantarum (1.6%), L. johnsonii (51.6%), L. fermentum (8.1%), L. salivarius (9.7%), L. reuter (1.6%), L. paracasei (8.1%), L. delbrueckii (3. 2% ) ; More than two different Lactobacillus species coexisted in 98% of women with vulvovaginal candidiasis, and no anyone species existed in 2% of them; In fertile women: L. iners (82.6%), L. cripatus (70.7%), L. gasseri (67.4%), L. jensenii (40.2%), L. acidophilus (39.1%), L. brevis (23.9%), L. plantarum (5.4%), L. rhamnosus (1.1%), L. paracasei (1.1%), L. reuter (1.1%) i, L. johnsonii (3.3%), L. fermentum (2.2%), L. salivarius (2.2%); More than two different Lactobacillus species coexisted in 97% of fertile women, and only one species existed in 3% of fertile women. Species of lactobacillus in women with vulvovaginal candidiasis did not significantly reduced compared with healthy women. Lactobacillus inert may be a marker of the change of vaginal microenvironment; Lactobacillus crispatus is a dominant lactobacillus in the vaginal of fertile healthy women, pregnant women and women with vulvovaginal candidiasis.

  4. A gastro-resistant ovalbumin bi-layered mini-tablet-in-tablet system for the delivery of Lactobacillus acidophilus probiotic to simulated human intestinal and colon conditions.

    PubMed

    Govender, Mershen; Choonara, Yahya Essop; van Vuuren, Sandy; Kumar, Pradeep; du Toit, Lisa Claire; Pillay, Viness

    2015-07-01

    The viability of probiotic bacteria during formulation processes and delivery is vital to ensure health benefits. This study focuses on the use of gastro-resistant denatured ovalbumin for the targeted delivery of probiotic Lactobacillus acidophilus to simulated human intestinal and colon conditions through a bi-layered mini-tablet-in-tablet system (BMTTS). The BMTTS consists of two gastro-resistant ovalbumin mini-tablets containing L. acidophilus suspended in lactose and eudragit S100 for targeted intestinal and colonic delivery respectively. Luminescence has been utilized to ensure probiotic viability during formulation processes in addition to determining all probiotic release profiles. The mechanism of probiotic release from the ovalbumin matrix was ascertained using mathematical modelling and molecular docking studies. Magnetic resonance imaging and differential scanning calorimetry are also included as part of the in-vitro characterization of the ovalbumin system. The BMTTS was effective in the delivery of L. acidophilus to simulated human intestinal and colon conditions. Formulation processes were furthermore determined to maintain probiotic viability. Statistical analysis of the release data noted a significant effect of pH denaturation on the release properties of ovalbumin. Magnetic resonance imaging results have indicated a decrease in ovalbumin matrix size upon exposure to simulated intestinal fluid. Molecular docking studies carried out depicted the interaction and binding positions inherent to the ovalbumin-pancreatic trypsin interaction complex indicating the possible enzymatic degradation of ovalbumin leading to the release of the probiotic from the protein matrix. The BMTTS has been determined to be effective in the protection and delivery of probiotic L. acidophilus to simulated human intestinal and colonic conditions. Molecular docking analysis has noted that pancreatin exerts a significant effect on probiotic release from the gastro

  5. The effects of Lactobacillus acidophilus as feed supplement on skin mucosal immune parameters, intestinal microbiota, stress resistance and growth performance of black swordtail (Xiphophorus helleri).

    PubMed

    Hoseinifar, Seyed Hossein; Roosta, Zahra; Hajimoradloo, Abdolmajid; Vakili, Farzaneh

    2015-02-01

    The present study evaluates the effects of different levels of dietary Lactobacillus acidophilus as feed supplement on intestinal microbiota, skin mucus immune parameters and salinity stress resistance as well as growth performance of black swordtail (Xiphophorus helleri). One-thousand and eight hundred healthy black swordtail larvae (0.03 ± 0.001 g) were randomly distributed in 12 tanks (100 L) at a density of 150 fish per aquaria and fed different levels of dietary L. acidophilus (0, 1.5 × 10(8), 3 × 10(8) and 6 × 10(8) CFU g(-1)) for 10 weeks. At the end of trial, there were significant differences among antibacterial activity of skin mucus in probiotic fed fish and control group (P < 0.05). Furthermore, the skin mucus protein level and alkaline phosphatase activity in control group were significantly lower than those of L. acidophilus fed fish (P < 0.05). Microbiological assessments revealed that feeding with probiotic supplemented diet remarkably increased total autochthonous bacteria and autochthonous lactic acid bacteria levels (P < 0.05). The results showed that dietary administration of L. acidophilus significantly elevated black swordtail resistance against salinity stress (i.e survival %) (P < 0.05). Also, dietary administration of different levels of L. acidophilus improved weight gain, SGR, FCR compared to fish fed unsupplemented diet (P < 0.05). These results demonstrate beneficial effects of dietary L. acidophilus on mucosal immune parameters, intestinal microbiota, stress resistance and growth parameters of black swordtail and the appropriate inclusion is 6 × 10(8) CFU g(-1). Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. Screening, Isolation and Identification of Probiotic Producing Lactobacillus acidophilus Strains EMBS081 & EMBS082 by 16S rRNA Gene Sequencing.

    PubMed

    Chandok, Harshpreet; Shah, Pratik; Akare, Uday Raj; Hindala, Maliram; Bhadoriya, Sneha Singh; Ravi, G V; Sharma, Varsha; Bandaru, Srinivas; Rathore, Pragya; Nayarisseri, Anuraj

    2015-09-01

    16S rDNA sequencing which has gained wide popularity amongst microbiologists for the molecular characterization and identification of newly discovered isolates provides accurate identification of isolates down to the level of sub-species (strain). Its most important advantage over the traditional biochemical characterization methods is that it can provide an accurate identification of strains with atypical phenotypic characters as well. The following work is an application of 16S rRNA gene sequencing approach to identify a novel species of Probiotic Lactobacillus acidophilus. The sample was collected from pond water samples of rural and urban areas of Krishna district, Vijayawada, Andhra Pradesh, India. Subsequently, the sample was serially diluted and the aliquots were incubated for a suitable time period following which the suspected colony was subjected to 16S rDNA sequencing. The sequence aligned against other species was concluded to be a novel, Probiotic L. acidophilus bacteria, further which were named L. acidophilus strain EMBS081 & EMBS082. After the sequence characterization, the isolate was deposited in GenBank Database, maintained by the National Centre for Biotechnology Information NCBI. The sequence can also be retrieve from EMBL and DDBJ repositories with accession numbers JX255677 and KC150145.

  7. Differential proteome and cellular adhesion analyses of the probiotic bacterium Lactobacillus acidophilus NCFM grown on raffinose - an emerging prebiotic.

    PubMed

    Celebioglu, Hasan Ufuk; Ejby, Morten; Majumder, Avishek; Købler, Carsten; Goh, Yong Jun; Thorsen, Kristian; Schmidt, Bjarne; O'Flaherty, Sarah; Abou Hachem, Maher; Lahtinen, Sampo J; Jacobsen, Susanne; Klaenhammer, Todd R; Brix, Susanne; Mølhave, Kristian; Svensson, Birte

    2016-05-01

    Whole cell and surface proteomes were analyzed together with adhesive properties of the probiotic bacterium Lactobacillus acidophilus NCFM (NCFM) grown on the emerging prebiotic raffinose, exemplifying a synbiotic. Adhesion of NCFM to mucin and intestinal HT-29 cells increased three-fold after culture with raffinose versus glucose, as also visualized by scanning electron microscopy. Comparative proteomics using 2D-DIGE showed 43 unique proteins to change in relative abundance in whole cell lysates from NCFM grown on raffinose compared to glucose. Furthermore, 14 unique proteins in 18 spots of the surface subproteome underwent changes identified by differential 2DE, including elongation factor G, thermostable pullulanase, and phosphate starvation inducible stress-related protein increasing in a range of +2.1 - +4.7 fold. By contrast five known moonlighting proteins decreased in relative abundance by up to -2.4 fold. Enzymes involved in raffinose catabolism were elevated in the whole cell proteome; α-galactosidase (+13.9 fold); sucrose phosphorylase (+5.4 fold) together with metabolic enzymes from the Leloir pathway for galactose utilization and the glycolysis; β-galactosidase (+5.7 fold); galactose (+2.9/+3.1 fold) and fructose (+2.8 fold) kinases. The insights at the molecular and cellular levels contributed to the understanding of the interplay of a synbiotic composed of NCFM and raffinose with the host. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  8. Development of Yoghurt with Juçara Pulp (Euterpe edulis M.) and the Probiotic Lactobacillus acidophilus La5.

    PubMed

    Geraldi, Marina V; Tulini, Fabricio L; Souza, Vanessa M; De Martinis, Elaine C P

    2018-03-01

    Yoghurts are dairy products consumed worldwide and can be supplemented with substances that provide extra health benefits as well as probiotic strains. In this context, the present study aimed to prepare a yoghurt added of juçara (Euterpe edulis M.) pulp and the commercial probiotic strain Lactobacillus acidophilus La5. Moreover, the probiotic survival during storage and after in vitro exposure to simulated gastric and enteric conditions was evaluated. Four formulations of yoghurt were prepared: (a) natural yoghurt, (b) yoghurt added of probiotic, (c) yoghurt added of juçara pulp, and (d) yoghurt added of probiotic culture and juçara pulp. The preparations were evaluated for survival of probiotic strain during storage and its tolerance to gastric and enteric conditions in vitro. The probiotic population in yoghurt remained unchanged during 28 days of storage. In addition, juçara pulp increased the probiotic resistance to simulated gastric and enteric conditions in the first day of storage. These data indicate that juçara pulp is a potential ingredient for the production of probiotic yoghurts.

  9. The other way around: Probiotic lactobacillus acidophilus NP51 restricts progression of Mycobacterium avium subspecies paratuberculosis (MAP) infection in Balb/c mice through activation of CD8+ T cell-mediated immunity

    USDA-ARS?s Scientific Manuscript database

    The objective of this study was to examine immune effects of feeding novel probiotic Lactobacillus acidophilus strain NP51 to specific pathogen-free Balb/c mice challenged with Mycobacterium avium subspecies paratuberculosis (MAP), the causative agent of Johne’s disease (JD). We hypothesized that fe...

  10. The effects of the DDS-1 strain of lactobacillus on symptomatic relief for lactose intolerance - a randomized, double-blind, placebo-controlled, crossover clinical trial.

    PubMed

    Pakdaman, Michael N; Udani, Jay K; Molina, Jhanna Pamela; Shahani, Michael

    2016-05-20

    Lactose intolerance is a form of lactose maldigestion where individuals experience symptoms such as diarrhea, abdominal cramping, flatulence, vomiting and bowel sounds following lactose consumption. Lactobacillus acidophilus is a species of bacteria known for its sugar fermenting properties. Preclinical studies have found that Lactobacillus acidophilus supplementation may assist in breaking down lactose; however, no human clinical trials exist evaluating its efficacy in alleviating symptoms related to lactose intolerance. The aim of this randomized, double-blind, placebo-controlled, crossover study was to evaluate the effect of a proprietary strain of Lactobacillus acidophilus on relieving discomfort related to lactose intolerance. The study enrolled healthy volunteers between 18 and 75 years of age who complained of lactose intolerance. Screening visits included a lactose challenge visit to confirm eligibility based on a score of 10 or higher on subjective assessment of the following symptoms after lactose challenge: diarrhea, abdominal cramping, vomiting, audible bowel sounds, flatulence, and overall symptoms. Qualified subjects participated in a 2-arm crossover design, with each arm consisting of 4 weeks of intervention of either active or placebo product, with a 2-week washout period during crossover. The study product consisted of the DDS-1 strain of Lactobacillus acidophilus (Nebraska Cultures, Walnut Creek, California). The placebo was formulated from maltodextrin. Study participants were instructed to take the product once daily for 4 weeks. Data collected included subjective symptom scores related to lactose intolerance. Longitudinal comparison between the DDS-1 group and placebo group demonstrated statistically significant reductions in abdominal symptom scores during the 6-h Lactose Challenge at week 4 for diarrhea (p = 0.033), abdominal cramping (p = 0.012), vomiting (p = 0.0002), and overall symptom score (p = 0.037). No adverse

  11. Effect of amino acid starvation on UV sensitivity of Lactobacillus acidophilus cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Soška, J.; Nečasová, J.

    1973-11-01

    In Lactobacillus acidophilus cultures uv irradiated in the exponential phase of growth, the dose-survival curve was of the simple exponential type, without any shoulder. If the bacteria were subjected to amino acid starvation prior to irradiation, a shoulder corresponding to a quasi-threshold dose (D) of about 780 ergs/mm/sup 2/ appeared in the curve. The administration of protein- or RNA-synthesis inhibitors prior to irradiation had the same effect. The effect of pre-irradiation amino acid starvation was abolished by simuitaneous thymidine starvation. It was likewise abolished if amino acid starvation was followed by incubation in the presence of amino acids (without thymidine)more » and then by irradiation of the cells. Post-irradiation amino acid starvation did not lead to the formation of a shoulder but if combined with thymidine starvation it did. It can be concluded from the results that post-irradiation repair processes are facilitated or promoted if, during the post-irradiation interval DNA synthesis is delayed. This delay represents a compensation of the pre-irradiation increase of cellular DNA-content, taking place during inhibition of proteosynthesis. The post-irradiation administration of caffeine did not abolish the formation of the shoulder induced by pre-irradiation amino acid starvation; on the contrary, it induced its formation even in exponentially growing, irradiated control bacteria. (auth)« less

  12. The Effect of Lactobacillus plantarum ATCC 8014 and Lactobacillus acidophilus NCFM Fermentation on Antioxidant Properties of Selected in Vitro Sprout Culture of Orthosiphon aristatus (Java Tea) as a Model Study

    PubMed Central

    Hunaefi, Dase; Akumo, Divine N.; Riedel, Heidi; Smetanska, Iryna

    2012-01-01

    High rosmarinic acid (RA) productivity has been achieved by applying jasmonic acid and yeast extract elicitors to the in vitro sprout culture of Orthosiphon aritatus (IOSC). The highest RA accumulation from three solvents was detected in IOSC after treatment with yeast extract (5 g/L). HPLC analysis clearly confirmed a drastic increase in RA subjected to yeast extract elicitation. Therefore, this yeast extract elicited IOSC was chosen for a lactic acid bacteria (LAB) fermentation study as a model system. This selected IOSC was subjected to different types of LAB fermentations (Lactobacillus plantarum ATCC 8014 and Lactobacillus acidophilus NCFM) for different periods of time 24, 48 and 72 h. The LAB fermentations consisted of solid state fermentations (SSF) and liquid state fermentations (LSF) in a Digital Control Unit (DCU) fermenter system. The aim was to determine the effect of fermentation on the antioxidant properties of the plant extract. Results indicated that all types of LAB fermentation decreased the level of RA and total phenolics, however, a slight increase in total flavonoids and flavonols was observed in SSF samples. HPLC results confirmed that the longer the fermentation, the greater the reduction in RA content. The highest reduction was obtained in the sample of LSF inoculated with L. plantarum for a period of 72 h. The temperature of fermentation (37 °C) was predicted as contributing to the declining level in RA content. The loss in RA was concomitant with a loss of total antioxidant activity (1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity, Trolox Equivalent Antioxidant Capacity (TEAC), and Superoxide Dismutase (SOD)-like activity). These results indicate that RA is the major contributor to the antioxidant activity of this plant. PMID:26787613

  13. A prebiotic mixture improved Lactobacillus acidophilus and Bifidobacterium animalis gastrointestinal in vitro resistance in petit-suisse.

    PubMed

    Padilha, Marina; Villarreal Morales, Martha Lissete; Vieira, Antonio Diogo Silva; Costa, Mayra Garcia Maia; Saad, Susana Marta Isay

    2016-05-18

    The survival of two probiotic strains -Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 - incorporated into probiotic (PC) and into synbiotic (SC, with inulin + fructooligosaccharides, respectively, at 7.5 and at 2.5 g per 100 g) petit-suisse cheese was investigated in the beginning (day 1) and at the end (28 days) of storage at 4 °C when the food products were subjected to in vitro gastrointestinal simulated assays. Species-specific quantitative real time PCR (qPCR) combined with propidium monoazide (PMA-qPCR) was employed to quantify the strains. Initial La-5 and Bb-12 populations were always above 7 log CFU g(-1). The presence of the prebiotic ingredients in SC improved the Bb-12 and La-5 resistance after the 6 h assay, with higher populations in all the in vitro stages and throughout the storage period (p < 0.05), leading to equal or superior survival rates (SR) in SC of both probiotic strains, in the beginning as well as at the end of storage. The mean La-5 SR were 58% (PC) and 67% (SC), whereas the mean Bb-12 SR were 60% (PC) and 79% (SC). Our findings suggest that the addition of a prebiotic mixture in petit-suisse cheese was advantageous, since it improved both the Bb-12 and La-5 viability and tolerance under in vitro gastrointestinal simulated conditions, both in the fresh product and in the product refrigerated for 28 days.

  14. Effect of probiotic strain Lactobacillus acidophilus (LBKV-3) on fecal residual lactase activity in undernourished children below 10 years.

    PubMed

    Hajare, Sunil Tulshiram; Bekele, Genene

    2017-01-01

    Clinically proven Lactobacillus acidophilus strain LBKV-3 intended as probiotic for humans was used to test its effect on fecal residual lactase activity in undernourished children below 10 years of age. The children were selected from malnutrition-declared area of Maharashtra (India). One of the major causes of malnutrition is lactose intolerance which leads to diarrhea. The basic consideration in selecting the probiotic strain of L. acidophilus (LBKV-3) in this investigation was the fact that the organism is isolated from human vaginal surface swab and it was found extensively studied for probiotic characteristic. LBKB 3 is tested by several workers as probiotic for hypocholesterolemic activity, implantation ability, therapeutic effects on gastrointestinal (GI) and related ailments. The results of present investigation have shown that the fecal residual lactase activity significantly increased than its initial value (which was almost zero). It appeared that the fecal residual β-galactosidase activity is an indication of positive implementation abilities of the cultures under investigation. These trends were compared with the control and blank group of children receiving Dahi and buffalo milk (BM). It was observed that both these products failed to exert any significant impact on increase in residual lactase activity.

  15. Nod2 is required for antigen-specific humoral responses against antigens orally delivered using a recombinant Lactobacillus vaccine platform

    PubMed Central

    Bumgardner, Sara A.; Zhang, Lin; LaVoy, Alora S.; Frank, Chad B.; Kajikawa, Akinobu; Klaenhammer, Todd R.

    2018-01-01

    Safe and efficacious orally-delivered mucosal vaccine platforms are desperately needed to combat the plethora of mucosally transmitted pathogens. Lactobacillus spp. have emerged as attractive candidates to meet this need and are known to activate the host innate immune response in a species- and strain-specific manner. For selected bacterial isolates and mutants, we investigated the role of key innate immune pathways required for induction of innate and subsequent adaptive immune responses. Co-culture of murine macrophages with L. gasseri (strain NCK1785), L. acidophilus (strain NCFM), or NCFM-derived mutants—NCK2025 and NCK2031—elicited an M2b-like phenotype associated with TH2 skewing and immune regulatory function. For NCFM, this M2b phenotype was dependent on expression of lipoteichoic acid and S layer proteins. Through the use of macrophage genetic knockouts, we identified Toll-like receptor 2 (TLR2), the cytosolic nucleotide-binding oligomerization domain containing 2 (NOD2) receptor, and the inflammasome-associated caspase-1 as contributors to macrophage activation, with NOD2 cooperating with caspase-1 to induce inflammasome derived interleukin (IL)-1β in a pyroptosis-independent fashion. Finally, utilizing an NCFM-based mucosal vaccine platform with surface expression of human immunodeficiency virus type 1 (HIV-1) Gag or membrane proximal external region (MPER), we demonstrated that NOD2 signaling is required for antigen-specific mucosal and systemic humoral responses. We show that lactobacilli differentially utilize innate immune pathways and highlight NOD2 as a key mediator of macrophage function and antigen-specific humoral responses to a Lactobacillus acidophilus mucosal vaccine platform. PMID:29734365

  16. The effect of addition of selected milk protein preparations on the growth of Lactobacillus acidophilus and physicochemical properties of fermented milk.

    PubMed

    Gustaw, Waldemar; Kozioł, Justyna; Radzki, Wojciech; Skrzypczak, Katarzyna; Michalak-Majewska, Monika; Sołowiej, Bartosz; Sławińska, Aneta; Jabłońska-Ryś, Ewa

    2016-01-01

    The intake of fermented milk products, especially yoghurts, has been systematically increasing for a few decades. The purpose of this work was to obtain milk products fermented with a mix of bacterial cultures (yoghurt bacteria and Lactobacillus acidophillus LA-5) and enriched with selected milk protein preparations. Secondly, the aim of the work was to determine physiochemical and rheological properties of the obtained products. The following additives were applied in the experiment: whey protein concentrate (WPC 65), whey protein isolate (WPI), demineralised whey powder (SPD), caseinoglycomacropeptide (CGMP), α-lactalbumin (α-la), sodium caseinate (KNa) and calcium caseinate (KCa). Milk was fermented using probiotic strain Lactobacillus acidophillus LA-5 and a typical yoghurt culture. The products were analysed in terms of the survivability of bacterial cells during refrigerated storage, rheological properties and syneresis. Fermented milk products were obtained using blends of bacterial strains: ST-B01:Lb-12 (1:1), ST-B01:Lb-12:LA-5 (1:1:2). Milk beverages fermented with typical yoghurt bacteria and LA-5 strain showed intensive syneresis. The addition of LA-5 strain caused formation of harder acid gels, comparing to typical yoghurts. Milk products which were prepared from skimmed milk possessed higher values of hardness and consistency coefficient. The increase of concentrations of milk preparations (except of WPI) did not cause significant differences in the hardness of acidic gels obtained by fermentation of mixed culture with a probiotic strain. The applied preparations improved physiochemical properties of the milk beverages which were prepared with a probiotic strain. The increase of protein milk preparations concentration resulted in a gradual decrease of the secreted whey. Among the products that were made of full milk powder and were subjected to three weeks of refrigerated storage the highest survivability of Lb. acidophilus LA-5 was noticed in the

  17. The other way around: probiotic Lactobacillus acidophilus NP51 restrict progression of Mycobacterium avium subspecies paratuberculosis (MAP) infection in Balb/c mice via activiation of CD8 alpha+ immune cell-mediated immunity

    USDA-ARS?s Scientific Manuscript database

    The objective of this study was to examine the immune-modulating effects of feeding a novel probiotic Lactobacillus acidophilus strain NP51 to specific pathogen-free Balb/c mice challenged with Mycobacterium avium subspecies paratuberculosis (MAP), the causative agent of Johne’s disease (JD) in rumi...

  18. Can Lactobacillus acidophilus improve minimal hepatic encephalopathy? A neurometabolite study using magnetic resonance spectroscopy.

    PubMed

    Ziada, Dina H; Soliman, Hanan H; El Yamany, Saher A; Hamisa, Manal F; Hasan, Azza M

    2013-09-01

    Minimal hepatic encephalopathy (MHE) is diagnosed when hepatic patients perform worse on psychometric tests compared to healthy controls. This study aimed to evaluate probiotics as alternative therapy in MHE. This is an open-label randomised controlled trial, performed in the Department of Tropical Medicine and Infectious Diseases, Tanta University Hospitals, from March 2010 to January 2012. A total of 90 patients with MHE were allocated by simple randomisation to three parallel equal groups. Group A received lactulose, group B a probiotic (Lactobacillus acidophilus) and group C served as the control. After informed consent, patients were tested for gut micrecology, fasting blood ammonia, liver functions and magnetic resonance spectroscopy (MRS) examination to study brain metabolites, mainly choline (Cho), myo-inositol (mI), glutamine+glutamate (Glx) and creatinin (Cre). Patients who developed overt encephalopathy were excluded from analysis. The whole battery of investigations was repeated in the same order after 4weeks. The probiotic was better tolerated than lactulose. The relative risk reduction (RRR) of developing overt encephalopathy was 60% in the case of lactulose and 80% in the case of probiotic, with a number needed to treat (NNT) of 2.4 and 2.3, respectively. The differential but not total microecology count was significantly shifted towards saccharolytic rather than proteolytic bacteria. The mI/Cre and (Cho+mI)/Glx ratios were significantly increased and the Glx/Cre ratio was significantly reduced after 1month-follow-up in the probiotic group compared to the lactulose group and in both treatment groups compared to the control group. Both probiotic and lactulose therapy can improve blood ammonia and psychometric tests in MHE and reduce the risk of developing overt encephalopathy. MRS showed more improvement in the levels of brain neurometabolites in the probiotic group. Copyright © 2013 Arab Journal of Gastroenterology. Published by Elsevier Ltd. All

  19. The Starvation Resistance and Biofilm Formation of Enterococcus faecalis in Coexistence with Candida albicans, Streptococcus gordonii, Actinomyces viscosus, or Lactobacillus acidophilus.

    PubMed

    Gao, Yan; Jiang, Xiaoqiong; Lin, Dongjia; Chen, Yanhuo; Tong, Zhongchun

    2016-08-01

    Enterococcus faecalis is the most frequently detected species in root canal-treated teeth, and it is able to survive under starvation conditions. However, persistent periapical disease is often caused by multispecies. The aim of this study was to explore the survival of E. faecalis in starvation conditions and biofilm formation with the 4 common pathogenic species. A dual-species model of Candida albicans, Streptococcus gordonii, Actinomyces viscosus, or Lactobacillus acidophilus in combination with E. faecalis was established and allowed to grow in phosphate-buffered saline for the examination of starvation survival. Cefuroxime sodium and vancomycin at a concentration of 100 mg/L were added into brain-heart infusion plate agar to count the 2 bacteria separately in the dual species. Scanning electron microscopy was used to observe the dual species and multiple species on the root canal dentin of bovine teeth for 48 hours. A confocal laser scanning microscope was used to show the 4 groups of dual-species biofilms on substrates with glass bottoms for 48 hours. E. faecalis was more resistant to starvation in coexistence with C. albicans, S. gordonii, A. viscosus, or L. acidophilus, and S. gordonii was completely inhibited in coexistence with E. faecalis. The dual-species biofilm showed that E. faecalis formed thicker and denser biofilms on the root canal dentin and glass slides in coexistence with S. gordonii and A. viscosus than C. albicans and L. acidophilus. The multispecies community is conducive to the resistance to starvation of E. faecalis and biofilm formation in root canals. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

  20. Stability of free and encapsulated Lactobacillus acidophilus ATCC 4356 in yogurt and in an artificial human gastric digestion system.

    PubMed

    Ortakci, F; Sert, S

    2012-12-01

    The objective of this study was to determine the effect of encapsulation on survival of probiotic Lactobacillus acidophilus ATCC 4356 (ATCC 4356) in yogurt and during artificial gastric digestion. Strain ATCC 4356 was added to yogurt either encapsulated in calcium alginate or in free form (unencapsulated) at levels of 8.26 and 9.47 log cfu/g, respectively, and the influence of alginate capsules (1.5 to 2.5mm) on the sensorial characteristics of yogurts was investigated. The ATCC 4356 strain was introduced into an artificial gastric solution consisting of 0.08 N HCl (pH 1.5) containing 0.2% NaCl or into artificial bile juice consisting of 1.2% bile salts in de Man, Rogosa, and Sharpe broth to determine the stability of the probiotic bacteria. When incubated for 2h in artificial gastric juice, the free ATCC 4356 did not survive (reduction of >7 log cfu/g). We observed, however, greater survival of encapsulated ATCC 4356, with a reduction of only 3 log cfu/g. Incubation in artificial bile juice (6 h) did not significantly affect the viability of free or encapsulated ATCC 4356. Moreover, statistically significant reductions (~1 log cfu/g) of both free and encapsulated ATCC 4356 were observed during 4-wk refrigerated storage of yogurts. The addition of probiotic cultures in free or alginate-encapsulated form did not significantly affect appearance/color or flavor/odor of the yogurts. However, significant deficiencies were found in body/texture of yogurts containing encapsulated ATCC 4356. We concluded that incorporation of free and encapsulated probiotic bacteria did not substantially change the overall sensory properties of yogurts, and encapsulation in alginate using the extrusion method greatly enhanced the survival of probiotic bacteria against an artificial human gastric digestive system. Copyright © 2012 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  1. Lactobacillus buchneri S-layer as carrier for an Ara h 2-derived peptide for peanut allergen-specific immunotherapy.

    PubMed

    Anzengruber, Julia; Bublin, Merima; Bönisch, Eva; Janesch, Bettina; Tscheppe, Angelika; Braun, Matthias L; Varga, Eva-Maria; Hafner, Christine; Breiteneder, Heimo; Schäffer, Christina

    2017-05-01

    Peanut allergy is an IgE-mediated severe hypersensitivity disorder. The lack of a treatment of this potentially fatal allergy has led to intensive research on vaccine development. Here, we describe the design and initial characterization of a carrier-bound peptide derived from the most potent peanut allergen, Ara h 2, as a candidate vaccine. Based on the adjuvant capability of bacterial surface (S-) layers, a fusion protein of the S-layer protein SlpB from Lactobacillus buchneri CD034 and the Ara h 2-derived peptide AH3a42 was produced. This peptide comprised immunodominant B-cell epitopes as well as one T cell epitope. The fusion protein SlpB-AH3a42 was expressed in E. coli, purified, and tested for its IgE binding capacity as well as for its ability to activate sensitized rat basophil leukemia (RBL) cells. The capacity of Ara h 2-specific IgG rabbit-antibodies raised against SlpB-AH3a42 or Ara h 2 to inhibit IgE-binding was determined by ELISA inhibition assays using sera of peanut allergic patients sensitized to Ara h 2. IgE specific to the SlpB-AH3a42 fusion protein was detected in 69% (25 of 36) of the sera. Despite the recognition by IgE, the SlpB-AH3a42 fusion protein was unable to induce β-hexosaminidase release from sensitized RBL cells at concentrations up to 100ng per ml. The inhibition of IgE-binding to the natural allergen observed after pre-incubation of the 20 sera with rabbit anti-SlpB-AH3a42 IgG was more than 30% for four sera, more than 20% for eight sera, and below 10% for eight sera. In comparison, anti-Ara h 2 rabbit IgG antibodies inhibited binding to Ara h 2 by 48% ±13.5%. Our data provide evidence for the feasibility of this novel approach towards the development of a peanut allergen peptide-based carrier-bound vaccine. Our experiments further indicate that more than one allergen-peptide will be needed to induce a broader protection of patients allergic to Ara h 2. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights

  2. Long-term multi-species Lactobacillus and Bifidobacterium dietary supplement enhances memory and changes regional brain metabolites in middle-aged rats.

    PubMed

    O'Hagan, Caroline; Li, Jia V; Marchesi, Julian R; Plummer, Sue; Garaiova, Iveta; Good, Mark A

    2017-10-01

    Ageing is associated with changes in the gut microbiome that may contribute to age-related changes in cognition. Previous work has shown that dietary supplements with multi-species live microorganisms can influence brain function, including induction of hippocampal synaptic plasticity and production of brain derived neurotrophic factor, in both young and aged rodents. However, the effect of such dietary supplements on memory processes has been less well documented, particularly in the context of aging. The main aim of the present study was to examine the impact of a long-term dietary supplement with a multi-species live Lactobacillus and Bifidobacteria mixture (Lactobacillus acidophilus CUL60, L. acidophilus CUL21, Bifidobacterium bifidum CUL20 and B. lactis CUL34) on tests of memory and behavioural flexibility in 15-17-month-old male rats. Following behavioural testing, the hippocampus and prefrontal cortex was extracted and analysed ex vivo using 1 H nuclear magnetic resonance ( 1 H NMR) spectroscopy to examine brain metabolites. The results showed a small beneficial effect of the dietary supplement on watermaze spatial navigation and robust improvements in long-term object recognition memory and short-term memory for object-in-place associations. Short-term object novelty and object temporal order memory was not influenced by the dietary supplement in aging rats. 1 H NMR analysis revealed diet-related regional-specific changes in brain metabolites; which indicated changes in several pathways contributing to modulation of neural signaling. These data suggest that chronic dietary supplement with multi-species live microorganisms can alter brain metabolites in aging rats and have beneficial effects on memory. Copyright © 2017 Elsevier Inc. All rights reserved.

  3. Functional and Probiotic Attributes of an Indigenous Isolate of Lactobacillus plantarum

    PubMed Central

    Kaushik, Jai K.; Kumar, Ashutosh; Duary, Raj K.; Mohanty, Ashok K.; Grover, Sunita; Batish, Virender K.

    2009-01-01

    Background Probiotic microorganisms favorably alter the intestinal microflora balance, promote intestinal integrity and mobility, inhibit the growth of harmful bacteria and increase resistance to infection. Probiotics are increasingly used in nutraceuticals, functional foods or in microbial interference treatment. However, the effectiveness of probiotic organism is considered to be population-specific due to variation in gut microflora, food habits and specific host-microbial interactions. Most of the probiotic strains available in the market are of western or European origin, and a strong need for exploring new indigenous probiotic organisms is felt. Methods and Findings An indigenous isolate Lp9 identified as Lactobacillus plantarum by molecular-typing methods was studied extensively for its functional and probiotic attributes, viz., acid and bile salt tolerance, cell surface hydrophobicity, autoaggregation and Caco-2 cell-binding as well as antibacterial and antioxidative activities. Lp9 isolate could survive 2 h incubation at pH 1.5–2.0 and toxicity of 1.5–2.0% oxgall bile. Lp9 could deconjugate major bile salts like glycocholate and deoxytaurocholate, indicating its potential to cause hypocholesterolemia. The isolate exhibited cell-surface hydrophobicity of ∼37% and autoaggregation of ∼31%. Presence of putative probiotic marker genes like mucus-binding protein (mub), fibronectin-binding protein (fbp) and bile salt hydrolase (bsh) were confirmed by PCR. Presence of these genes suggested the possibility of specific interaction and colonization potential of Lp9 isolate in the gut, which was also suggested by a good adhesion ratio of 7.4±1.3% with Caco-2 cell line. The isolate demonstrated higher free radical scavenging activity than standard probiotics L. johnsonii LA1 and L. acidophilus LA7. Lp9 also exhibited antibacterial activity against E. coli, L. monocytogenes, S. typhi, S. aureus and B. cereus. Conclusion The indigenous Lactobacillus plantarum Lp

  4. Microencapsulation of Lactobacillus acidophilus NCIMB 701748 in matrices containing soluble fibre by spray drying: Technological characterization, storage stability and survival after in vitro digestion☆

    PubMed Central

    Yonekura, Lina; Sun, Han; Soukoulis, Christos; Fisk, Ian

    2014-01-01

    We evaluated sodium alginate, chitosan and hydroxypropyl methylcellulose (HPMC) as co-encapsulants for spray dried Lactobacillus acidophilus NCIMB 701748 by assessing their impact on cell viability and physicochemical properties of the dried powders, viability over 35 days of storage at 25 °C and survival after simulated digestion. Fibres were added to a control carrier medium containing whey protein concentrate, d-glucose and maltodextrin. Sodium alginate and HPMC did not affect cell viability but chitosan reduced viable counts in spray dried powders, as compared to the control. Although chitosan caused large losses of viability during spray-drying, these losses were counteracted by the excellent storage stability compared to control, sodium alginate and HPMC, and the overall effect became positive after the 35-day storage. Chitosan also improved survival rates in simulated GI conditions, however no single fibre could improve L. acidophilus NCIMB 701748 viability in all steps from production through storage and digestion. PMID:24748900

  5. Genome Sequence of the Enterohemorrhagic Escherichia coli Bacteriophage UFV-AREG1

    PubMed Central

    Batalha, Laís Silva; Albino, Luiz Augusto A.; Boggione, Delaine Meireles Gouveia; Gontijo, Marco Tulio Pardini; Bazzolli, Denise M. Soares; Mendonca, Regina C. Santos

    2016-01-01

    Here, we present the genome sequence of the Escherichia coli bacteriophage UFV-AREG1. This phage was isolated from cowshed wastewater and showed specificity for enterohemorrhagic E. coli O157:H7 (ATCC 43895), E. coli 0111 (CDC O11ab) and E. coli (ATCC 23229). PMID:27738021

  6. Salivary mutans streptococci and lactobacilli modulations in young children on consumption of probiotic ice-cream containing Bifidobacterium lactis Bb12 and Lactobacillus acidophilus La5.

    PubMed

    Singh, Richa Polka; Damle, Satyawan Gangaram; Chawla, Amrita

    2011-11-01

    To compare the levels of mutans streptococci and lactobacilli in saliva of school children, before and after consumption of probiotic and control ice-cream. A double-blind, cross-over, placebo-controlled trial was carried out in forty, 12-14 year-old children, with no clinically detectable caries. The selected children were randomized equally into two groups I and II. Following an initial run-in period of 1 week, children in group I and II were given ice-creams 'A' and 'B', respectively, for 10 days. Being a cross-over study, the ice-creams were interchanged in the two groups after a 2-week wash-out period. Saliva samples at baseline and follow-up were assessed using Dentocult SM and Dentocult LB kits. On statistical evaluation, it was seen that probiotic ice-cream brought about a statistically significant reduction (p-value = 0.003) in salivary mutans streptococci levels with no significant effect on lactobacilli levels. In conclusion, probiotic ice-cream containing Bifidobacterium lactis Bb-12 ATCC27536 and Lactobacillus acidophilus La-5 can reduce the levels of certain caries-associated micro-organisms in saliva.

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

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    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).

  8. Live encapsulated Lactobacillus acidophilus cells in yogurt for therapeutic oral delivery: preparation and in vitro analysis of alginate-chitosan microcapsules.

    PubMed

    Urbanska, Aleksandra Malgorzata; Bhathena, Jasmine; Prakash, Satya

    2007-09-01

    Targeted delivery of live microencapsulated bacterial cells has strong potential for application in treating various diseases, including diarrhea, kidney failure, liver failure, and high cholesterol, among others. This study investigates the potential of microcapsules composed of two natural polymers, alginate and chitosan (AC), and the use of these artificial cells in yogurt for delivery of probiotic Lactobacillus acidophilus bacterial live cells. Results show that the integrity of AC microcapsules was preserved after 76 h of mechanical shaking in MRS broth and after 12 h and 24 h in simulated gastric and intestinal fluids. Using an in vitro computer-controlled simulated human gastrointestinal (GI) model, we found 8.37 log CFU/mL of viable bacterial cells were present after 120 min of gastric exposure and 7.96 log CFU/mL after 360 min of intestinal exposure. In addition, AC microcapsules composed of chitosan 10 and 100 at various concentrations were subjected to 4-week storage in 2% milk fat yogurt or 0.85% physiological solution. It was found that 9.37 log CFU/mL of cells encapsulated with chitosan 10 and 8.24 log CFU/mL of cells encapsulated with chitosan 100 were alive after 4 weeks. The AC capsule composed of 0.5% chitosan 10 provided the highest bacterial survival of 9.11 log CFU/mL after 4 weeks. Finally, an investigation of bacterial viability over 72 h in different pH buffers yielded highest survival of 6.34 log CFU/mL and 10.34 log CFU/mL at pH 8 for free and AC-encapsulated cells, respectively. We conclude from these findings that encapsulation allows delivery of a higher number of bacteria to desired targets in the GI tract and that microcapsules containing bacterial cells are good candidates for oral artificial cells for bacterial cell therapy.

  9. The effect of probiotic bacteria (Lactobacillus acidophilus and Bifidobacterium lactis) on the accumulation of lead in rat brains.

    PubMed

    Zanjani, Saman Yahyavi; Eskandari, Mohammad Reza; Kamali, Koorosh; Mohseni, Mehran

    2017-01-01

    Lead is a toxic metal present in different concentrations in a wide variety of food products. Exposure to lead, even to low levels, causes acute and chronic toxicities. Lead can cross the blood-brain barrier and accumulate in the nervous system. Probiotics are live microorganisms that, when used in adequate amounts, confer a health benefit on the host. Although a recent study demonstrated that the studied bacteria have a protective effect against acute lead toxicity, no research has been found that shows the long-term impact of these bacteria in vivo. The current study surveyed the protective effects of two species of probiotics, Lactobacillus acidophilus LA-5 and Bifidobacterium lactis BB-12, that are most widely used in many functional foods against oral lead exposure (4 weeks) in rat brains. The results revealed that, at the end of the second week of chronic exposure to lead and probiotic bacteria, the lowest level of lead belonged to the Lactobacillus group. At the end of the fourth week, the lowest amount of lead was related to the group receiving both types of probiotics. With the physiological benefits of probiotic consumption, the bacterial solution in this study did not show high efficacy in reducing brain lead concentrations.

  10. Two Lactobacillus Species Inhibit the Growth and α-Toxin Production of Clostridium perfringens and Induced Proinflammatory Factors in Chicken Intestinal Epithelial Cells in Vitro

    PubMed Central

    Guo, Shuangshuang; Liu, Dan; Zhang, Beibei; Li, Zhui; Li, Yehan; Ding, Binying; Guo, Yuming

    2017-01-01

    Clostridium perfringens is the causative pathogen of avian necrotic enteritis. Lactobacillus spp. are well-characterized probiotics with anti-microbial and immune-modulatory activities. In the present study, we investigated the effects of L. acidophilus and L. fermentum on the growth, α-toxin production and inflammatory responses of C. perfringens. In in vitro culture experiments, both lactobacilli inhibited the growth of C. perfringens (P < 0.01), accompanied with a decrease in pH (P < 0.01). Supernatants from lactobacilli cultures also suppressed the growth of C. perfringens during 24 h of incubation (P < 0.01), but this inhibitory effect disappeared after 48 h. Both lactobacilli decreased the α-toxin production of C. perfringens (P < 0.01) without influencing its biomass, and even degraded the established α-toxin (P < 0.01). Lower environmental pH reduced the α-toxin production as well (P < 0.01). Preincubation with L. acidophilus decreased the attachment of C. perfringens to cells (P < 0.01) with the cell cytotoxicity being unaffected. Both lactobacilli pretreatment reduced the up-regulation of proinflammatory factors, peptidoglycan (PGN) receptors and nuclear factor kappa B (NF-κB) p65 in C. perfringens-challenged chicken intestinal epithelial cells (P < 0.05). In conclusion, L. acidophilus and L. fermentum inhibited the pathological effects of C. perfringens in vitro conditions. PMID:29118744

  11. Detection and Identification of Lactobacillus Species in Crops of Broilers of Different Ages by Using PCR-Denaturing Gradient Gel Electrophoresis and Amplified Ribosomal DNA Restriction Analysis

    PubMed Central

    Guan, Le Luo; Hagen, Karen E.; Tannock, Gerald W.; Korver, Doug R.; Fasenko, Gaylene M.; Allison, Gwen E.

    2003-01-01

    The microflora of the crop was investigated throughout the broiler production period (0 to 42 days) using PCR combined with denaturing gradient gel electrophoresis (PCR-DGGE) and selective bacteriological culture of lactobacilli followed by amplified ribosomal DNA restriction analysis (ARDRA). The birds were raised under conditions similar to those used in commercial broiler production. Lactobacilli predominated and attained populations of 108 to 109 CFU per gram of crop contents. Many of the lactobacilli present in the crop (61.9% of isolates) belonged to species of the Lactobacillus acidophilus group and could not be differentiated by PCR-DGGE. A rapid and simple ARDRA method was developed to distinguish between the members of the L. acidophilus group. HaeIII-ARDRA was used for preliminary identification of isolates in the L. acidophilus group and to identify Lactobacillus reuteri and Lactobacillus salivarius. MseI-ARDRA generated unique patterns for all species of the L. acidophilus group, identifying Lactobacillus crispatus, Lactobacillus johnsonii, and Lactobacillus gallinarum among crop isolates. The results of our study provide comprehensive knowledge of the Lactobacillus microflora in the crops of birds of different ages using nucleic acid-based methods of detection and identification based on current taxonomic criteria. PMID:14602636

  12. Efficacy of prolonged ingestion of Lactobacillus acidophilus L-92 in adult patients with atopic dermatitis.

    PubMed

    Yamamoto, Kozo; Yokoyama, Kazuhito; Matsukawa, Takehisa; Kato, Sayaka; Kato, Shinji; Yamada, Kazuhisa; Hirota, Tatsuhiko

    2016-07-01

    To evaluate the safety and efficacy of prolonged ingestion of Lactobacillus acidophilus L-92 (L-92) on skin symptoms in adult atopic dermatitis (AD) patients, a placebo-controlled double-blinded parallel-group comparison study was performed. This included daily administration of heat-killed and dried L-92 or placebo for 24wk in 50 AD patients who were 16yr old or older. The severity of skin symptoms was evaluated at baseline and at 4, 8, 12, 16, 20, and 24wk during the intervention using the investigator global assessment, eczema area and severity index, and scoring atopic dermatitis. Serum cytokine and blood marker levels were also measured at baseline and at 4, 8, 16, and 24wk during the intervention. No adverse events were reported during the study period. Compared with the placebo group, the L-92 group showed significant decreases in investigator global assessment, eczema area and severity index, and scoring atopic dermatitis. Subjective symptoms in adult AD patients were reduced by intake of L-92. Furthermore, it was suggested that sustained ingestion of L-92 resulted in suppression of scratching behavior and maintenance of remission status of skin symptoms. Sixteen weeks after the study commenced, a significant decrease in lactate dehydrogenase and a significant increase in transforming growth factor-β were observed in the L-92 group compared with the placebo group. In the L-92 group, a significant elevation of IL-12 (p70) level at the end of treatment period compared with before the treatment was observed. This study suggested that L-92 suppresses type-2-helper-T-cell-dominant inflammation by activating regulatory T cells and type 1 helper T cells. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  13. Probiotics (Lactobacillus rhamnosus R0011 and acidophilus R0052) Reduce the Expression of Toll-Like Receptor 4 in Mice with Alcoholic Liver Disease

    PubMed Central

    Hong, Meegun; Kim, Seung Woo; Han, Sang Hak; Kim, Dong Joon; Suk, Ki Tae; Kim, Yeon Soo; Kim, Myong Jo; Kim, Moon Young; Baik, Soon Koo; Ham, Young Lim

    2015-01-01

    Objective The role of lipopolysaccharide (LPS) and toll-like receptor 4 (TLR 4) in the pathogenesis of alcoholic liver disease (ALD) has been widely established. We evaluated the biological effects of probiotics (Lactobacillus rhamnosus R0011 and acidophilus R0052), KRG (Korea red ginseng), and urushiol (Rhus verniciflua Stokes) on ALD, including their effects on normal and high-fat diet in mice. Methods One hundred C57BL/6 mice were classified into normal (N) and high-fat diet (H) groups. Each group was divided into 5 sub-groups: control, alcohol, alcohol+probiotics, alcohol+KRG, and alcohol+urushiol. A liver function test, histology, electron-microscopy, interleukin (IL)-1β, tumor necrosis factor (TNF)-α, IL-6, and IL-10, and TLR 4 were evaluated and compared. Results In the N group, probiotics, KRG, and urushiol significantly reduced levels of TNF-α (12.3±5.1, 13.4±3.9, and 12.1±4.3 vs. 27.9±15.2 pg/mL) and IL-1β (108.4±39.4, 75.0±51.0, and 101.1±26.8 vs. 162.4±37.5 pg/mL), which were increased by alcohol. Alcohol-induced TLR 4 expression was reduced by probiotics and urushiol (0.7±0.2, and 0.8±0.1 vs. 1.0±0.3, p<0.001). In the H group, IL-10 was significantly increased by probiotics and KRG, compared with alcohol (25.3±15.6 and 20.4±6.2 vs. 7.6±5.6 pg/mL) and TLR 4 expression was reduced by probiotics (0.8±0.2 vs. 1.0±0.3, p = 0.007). Conclusions Alcohol-induced TLR 4 expression was down-regulated by probiotics in the normal and high-fat diet groups. Probiotics, KRG, and urushiol might be effective in the treatment of ALD by regulating the gut-liver axis. PMID:25692549

  14. Identification and Characterization of Novel Surface Proteins in Lactobacillus johnsonii and Lactobacillus gasseri

    PubMed Central

    Ventura, Marco; Jankovic, Ivana; Walker, D. Carey; Pridmore, R. David; Zink, Ralf

    2002-01-01

    We have identified and sequenced the genes encoding the aggregation-promoting factor (APF) protein from six different strains of Lactobacillus johnsonii and Lactobacillus gasseri. Both species harbor two apf genes, apf1 and apf2, which are in the same orientation and encode proteins of 257 to 326 amino acids. Multiple alignments of the deduced amino acid sequences of these apf genes demonstrate a very strong sequence conservation of all of the genes with the exception of their central regions. Northern blot analysis showed that both genes are transcribed, reaching their maximum expression during the exponential phase. Primer extension analysis revealed that apf1 and apf2 harbor a putative promoter sequence that is conserved in all of the genes. Western blot analysis of the LiCl cell extracts showed that APF proteins are located on the cell surface. Intact cells of L. johnsonii revealed the typical cell wall architecture of S-layer-carrying gram-positive eubacteria, which could be selectively removed with LiCl treatment. In addition, the amino acid composition, physical properties, and genetic organization were found to be quite similar to those of S-layer proteins. These results suggest that APF is a novel surface protein of the Lactobacillus acidophilus B-homology group which might belong to an S-layer-like family. PMID:12450842

  15. Effect of Type of Protein-Based Microcapsules and Storage at Various Ambient Temperatures on the Survival and Heat Tolerance of Spray Dried Lactobacillus acidophilus.

    PubMed

    Dianawati, Dianawati; Lim, Seng Feng; Ooi, Yasmin Beng Houi; Shah, Nagendra P

    2017-09-01

    The aims of this study were to evaluate the effect of types of protein-based microcapsules and storage at various ambient temperatures on the survival of Lactobacillus acidophilus during exposure to simulated gastrointestinal tract and on the change in thermo-tolerance during heating treatment. The encapsulating materials were prepared using emulsions of protein (sodium caseinate, soy protein isolate, or pea protein), vegetable oil, and glucose, with maltodextrin was used as a wall material. The formulations were heated at 90 °C for 30 min to develop Maillard substances prior to being incorporated with L. acidophilus. The mixtures were then spray dried. The microspheres were stored at 25, 30, and 35 °C for 8 wk and examined every 4 wk. The addition of proteins as encapsulating materials demonstrated a significant protective effect (P < 0.05) as compared to the control sample. Sodium caseinate and soy protein isolate appeared more effective than pea protein in protecting the bacteria after spray drying and during the storage at different room temperatures. Storage at 35 °C resulted in a significant decrease in survival at end of storage period regardless the type of encapsulating materials. The addition of protein-based materials also enhanced the survival of L. acidophilus during exposure to simulated gastrointestinal condition as compared to the control. After spray drying and after 0th wk storage, casein, soy protein isolate, and pea protein-based formulations protected the bacteria during heat treatment. In fact, a significant decrease in thermal tolerance was inevitable after 2 wk of storage at 25 °C. © 2017 Institute of Food Technologists®.

  16. RNF20 and USP44 regulate stem cell differentiation by modulating H2B monoubiquitylation

    PubMed Central

    Fuchs, Gilad; Shema, Efrat; Vesterman, Rita; Kotler, Eran; Wolchinsky, Zohar; Wilder, Sylvia; Golomb, Lior; Pribluda, Ariel; Zhang, Feng; Haj-Yahya, Mahmood; Feldmesser, Ester; Brik, Ashraf; Yu, Xiaochun; Hanna, Jacob; Aberdam, Daniel; Domany, Eytan; Oren, Moshe

    2012-01-01

    Summary Embryonic stem cells (ESC) maintain high genomic plasticity, essential for their capacity to enter diverse differentiation pathways. Post-transcriptional modifications of chromatin histones play a pivotal role in maintaining this plasticity. We now report that one such modification, monoubiquitylation of histone H2B on lysine 120 (H2Bub1), catalyzed by the E3 ligase RNF20, increases during ESC differentiation and is required for efficient execution of this process. This increase is particularly important for the transcriptional induction of relatively long genes during ESC differentiation. Furthermore, we identify the deubiquitinase USP44 as a negative regulator of H2B ubiquitylation, whose downregulation during ESC differentiation contributes to the increase in H2Bub1. Our findings suggest that optimal ESC differentiation requires dynamic changes in H2B ubiquitylation patterns, which must occur in a timely and well-coordinated manner. PMID:22681888

  17. Lactobacillus acidophilus alleviates pouchitis after ileal pouch-anal anastomosis in rats.

    PubMed

    Xu, Yan-Yan; Zhang, Ying-Ying; He, An-Qi; Li, Kai-Yu; Gao, Sen-Yang; Liu, Gang

    2017-07-14

    To assess the therapeutic potential of Lactobacillus acidophilus (LA) for the treatment of pouchitis in a rat model. Sprague Dawley rats underwent proctocolectomy and ileal pouch-anal anastomosis followed by administration of dextran sulfate sodium (DSS) to induce pouchitis. Rats with pouchitis were randomly divided into three groups: no intervention (NI), normal saline (NS, 3 mL/d normal saline for 7 d), and LA (3 mL/d LA at 1× 10 10 colony-forming units for 7 d). General body condition was recorded and pouch specimens were obtained for histological examination. mRNA expression levels of interleukin (IL)-1β, IL-6, IL-10, and tumor necrosis factor-α were determined by RT-PCR. Zonula occludens protein 1 (ZO-1) levels were measured by immunohistochemistry. LA reduced weight loss associated with pouchitis ( P < 0.05) and improved the symptoms of pouchitis in rats. Compared with the NI and NS groups, rats in the LA group showed earlier disappearance of hematochezia (6.17 ± 0.75, 6.50 ± 0.55, 3.17 ± 0.75, P < 0.05) and higher fecal scores (2.67 ± 0.48, 2.50 ± 0.51, 4.42 ± 0.50, respectively, P < 0.05). Histological scores were also lower in the LA group compared with the other two groups (7.17 ± 0.98, 8.00 ± 0.89, 4.00 ± 0.89, respectively, P < 0.05). mRNA expression levels of IL-1β, IL-6, and tumor necrosis factor-α were significantly reduced, while IL-10 mRNA levels were significantly increased in the LA group ( P < 0.05, respectively). ZO-1 protein levels were also significantly increased after administration of LA ( P < 0.05). LA alleviates pouchitis induced by DSS after ileal pouch-anal anastomosis by decreasing pro-inflammatory factors and increasing anti-inflammatory factors, and restoring ZO-1 expression in the mucosa.

  18. Eradication of Helicobacter pylori infection by the probiotic strains Lactobacillus johnsonii MH-68 and L. salivarius ssp. salicinius AP-32.

    PubMed

    Hsieh, Pei-Shan; Tsai, Yi-Chun; Chen, Yi-Chun; Teh, Su-Fen; Ou, Chung-Mou; King, V An-Erl

    2012-12-01

    The current therapy for Helicobacter pylori infection includes antimicrobial agents and proton pump inhibitors. We have examined the ability of Lactobacillus spp. to inhibit H. pylori infection. Probiotic strains isolated from samples of adult feces, infant feces, breast milk, and vaginal swab collected from healthy volunteers in Taiwan and commercially available strains were screened for antagonism toward H. pylori. Inhibition liquid culture assay was used to screen potential anti-H. pylori activity. Then, we performed agar plate inhibition assay, and assays to determine the capacity of probiotics for adhesion, and inhibition and killing of H. pylori, and measured the levels of IL-8 and IL-10. Using animal models, we studied regulation of gastric acid and histopathological changes accompanying anti-H. pylori activity. We found that six of the tested strains suppressed urease activity of H. pylori: Lactobacillus acidophilus TYCA08, L. acidophilus TYCA15, L. johnsonii MH-68, and L. salivarius subsp. salicinius AP-32 were more effective than the others. In vivo, L. johnsonii MH-68 and L. salivarius subsp. salicinius AP-32 alone or in combination, reduced the H. pylori load in the gastric mucosa, and also reduced inflammatory chemokine expression and lymphocyte infiltration. Lactobacillus johnsonii MH-68 and L. salivarius subsp. salicinius AP-32 effectively suppress H. pylori viability, and when used as probiotics, they may help decrease the occurrence of gastritis, and even reduce the risk of H. pylori infection. © 2012 Blackwell Publishing Ltd.

  19. Salmonella enterica serovar typhimurium colonization of the crop in the domestic turkey: influence of probiotic and prebiotic treatment (Lactobacillus acidophilus and lactose).

    PubMed

    Johannsen, Sara A; Griffith, Ronald W; Wesley, Irene V; Scanes, Colin G

    2004-01-01

    Acute colonization of the crop of the domestic turkey by Salmonella enterica serovar typhimurium (ST) was examined. The influences of preharvest probiotic and prebiotic treatment with lactobaccilli and lactose on crop colonization with ST were also investigated. Prior to Salmonella challenge, poults received 2.5% lactose and Lactobacillus acidophilus (1.9 x 10(9) organisms/liter) in the only source of drinking water from 1 day old to termination. At 3-wk-old, turkey poults were challenged with ST (1.7 X 10(8) colony-forming units [CFU]/ml) before their natural nocturnal fast to determine the potential effects of supplementation on crop colonization when the crop was engorged and subsequently undergoing emptying. Crop ingesta and tissue were collected at time points 30 min and 4, 8, and 24 hr postchallenge and ST levels were determined. High levels of ST were detected in the crop. For instance, for the poults not receiving lactose or lactobacilli, 30 min after ST challenge, there were 4.4 x 10(7) CFU in the crop ingesta and 5.3 x 10(5) CFU in the crop wall. Ingesta ST levels dropped dramatically to 1.0 x 10(6) CFU after 4 hr as the crop emptied. Crop wall ST levels were steady during the nocturnal crop evacuation. Immunohistochemical staining demonstrated ST in close association with the crop epithelium. Treatment with lactose and L. acidophilus supplementation did not reduce ST colonization.

  20. Effect of metronidazole supplemented with hydroquinone on the adhesion of Lactobacillus acidophilus in ovine vaginal cells.

    PubMed

    Coletti Zabala, Tamara L; Zerbatto, María E; Perotti, Elda B R; Smacchia, Ana M; Ombrella, Adriana; Pidello, Alejandro R

    This work demonstrates that the addition of metronidazole together with a ubiquitous quinone compound reduces adherence of Lactobacillus acidophilus to ovine vaginal cells. Spectrophotometric and voltammetric studies have shown that neoformed compounds were observed in these systems; there were also changes in their electroactive composition, and the oxidant status had a significantly higher value compared to the control (p<0.05). Based on reduction potential (E; mV), the distribution of electroactive compound concentrations suggests that the compounds with low reduction potential induce this behavior, which would indicate that the addition of metronidazole with a ubiquitous quinone compound to the vaginal system might increase the reductive capacity of these systems. This work shows that the study of behavior and fluctuations of the redox compounds that compose the vaginal environment, in terms of concentration and species of redox molecules, must be hierarchized in order to better understand the early stages of colonization by microorganisms. Copyright © 2016 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  1. Probiotic Strawberry Yogurts: Microbiological, Chemical and Sensory Properties.

    PubMed

    Turgut, Tamer; Cakmakci, Songül

    2018-03-01

    This study was performed to determine the viability of Lactobacillus acidophilus and Bifidobacterium bifidum in yogurt made with strawberry marmalade (SM) and to examine the quality properties of probiotic yogurt. Acidity, pH, bacterial counts and sensory analysis of the yogurt samples were investigated on days 1, 3, 5, 7, 10 and 14 during storage at 4 °C. The survival rate of L. acidophilus was greater than that of B. bifidum. The viability of L. acidophilus decreased during the storage period, but B. bifidum numbers remained stable during the storage period. The highest L. acidophilus count (7.20 log cfu/g) was found in L. acidophilus + B. bifidum SM yogurt on day 1. The highest B. bifidum count (6.13 log cfu/g) was detected in yogurt containing L. acidophilus + B. bifidum SM yogurt on day 7. Yeast and mould counts of all yogurts increased during the storage period. Coliform bacteria and Staphylococcus aureus were not detected in the yogurt samples. The highest overall acceptance sensory score was observed in yogurts containing L. acidophilus. Considering the sensory and probiotic characteristics of all yogurt samples, this study suggested that strawberry yogurt with a suitable 5-7 day storage period can be produced with single L. acidophilus addition or single B. bifidum addition.

  2. In vitro gastrointestinal resistance of Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 in soy and/or milk-based synbiotic apple ice creams.

    PubMed

    Matias, Natalia Silva; Padilha, Marina; Bedani, Raquel; Saad, Susana Marta Isay

    2016-10-03

    The viability and resistance to simulated gastrointestinal (GI) conditions of Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 in synbiotic ice creams, in which milk was replaced by soy extract and/or whey protein isolate (WPI) with inulin, were investigated. The ice creams were showed to be satisfactory vehicles for La-5 and Bb-12 (populations around 7.5logCFU/g), even after the whole storage period (84days/-18°C). In all formulations, the propidium monoazide qPCR (PMA-qPCR) analysis demonstrated that probiotics could resist the in vitro GI assay, with significant survival levels, achieving survival rates exceeding 50%. Additionally, scanning electron microscopy images evidenced cells with morphological differences, suggesting physiological changes in response to the induced stress during the in vitro assay. Although all formulations provided resistance to the probiotic strains under GI stress, the variation found in probiotic survival suggests that GI tolerance is indeed affected by the choice of the food matrix. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. Effects of synbiotic fermented milk containing Lactobacillus acidophilus La-5 and Bifidobacterium animalis ssp. lactis BB-12 on the fecal microbiota of adults with irritable bowel syndrome: A randomized double-blind, placebo-controlled trial.

    PubMed

    Bogovič Matijašić, Bojana; Obermajer, Tanja; Lipoglavšek, Luka; Sernel, Tjaša; Locatelli, Igor; Kos, Mitja; Šmid, Alenka; Rogelj, Irena

    2016-07-01

    We conducted a randomized double-blind, placebo-controlled multicentric study to investigate the influence of a synbiotic fermented milk on the fecal microbiota composition of 30 adults with irritable bowel syndrome (IBS). The synbiotic product contained Lactobacillus acidophilus La-5, Bifidobacterium animalis ssp. lactis BB-12, Streptococcus thermophilus, and dietary fiber (90% inulin, 10% oligofructose), and a heat-treated fermented milk without probiotic bacteria or dietary fiber served as placebo. Stool samples were collected after a run-in period, a 4-wk consumption period, and a 1-wk follow-up period, and were subjected to real-time PCR and 16S rDNA profiling by next-generation sequencing. After 4wk of synbiotic (11 subjects) or placebo (19 subjects) consumption, a greater increase in DNA specific for L. acidophilus La-5 and Bifidobacterium animalis ssp. lactis was detected in the feces of the synbiotic group compared with the placebo group by quantitative real-time PCR. After 1wk of follow-up, the content of L. acidophilus La-5 and B. animalis ssp. lactis decreased to levels close to initial levels. No significant changes with time or differences between the groups were observed for Lactobacillus, Enterobacteriaceae, Bifidobacterium, or all bacteria. The presence of viable BB-12- and La-5-like bacteria in the feces resulting from the intake of synbiotic product was confirmed by random amplification of polymorphic DNA (RAPD)-PCR. At the end of consumption period, the feces of all subjects assigned to the synbiotic group contained viable bacteria with a BB-12-like RAPD profile, and after 1wk of follow-up, BB-12-like bacteria remained in the feces of 87.5% of these subjects. The presence of La-5-like colonies was observed less frequently (37.5 and 25% of subjects, respectively). Next-generation sequencing of 16S rDNA amplicons revealed that only the percentage of sequences assigned to Strep. thermophilus was temporarily increased in both groups, whereas the

  4. The Copper Homeostasis Transcription Factor CopR Is Involved in H2O2 Stress in Lactobacillus plantarum CAUH2

    PubMed Central

    Yang, Yang; Yin, Jia; Liu, Jie; Xu, Qi; Lan, Tian; Ren, Fazheng; Hao, Yanling

    2017-01-01

    Transcriptional factors (TFs) play important roles in the responses to oxidative, acid, and other environmental stresses in Gram-positive bacteria, but the regulatory mechanism of TFs involved in oxidative stress remains unknown in lactic acid bacteria. In the present work, homologous overexpression strains with 43 TFs were constructed in the Lactobacillus plantarum CAUH2 parent strain. The strain overexpressing CopR displayed the highest sensitivity and a 110-fold decrease in survival rate under H2O2 challenge. The importance of CopR in the response to H2O2 stress was further confirmed by a 10.8-fold increase in the survival of a copR insertion mutant. In silico analysis of the genes flanking copR revealed putative CopR-binding “cop box” sequences in the promoter region of the adjacent gene copB encoding a Cu2+-exporting ATPase. Electrophoretic mobility shift assay (EMSA) analysis demonstrated the specific binding of CopR with copB in vitro, suggesting copB is a target gene of CopR in L. plantarum. The role of CopB involved in oxidative stress was verified by the significantly decreased survival in the copB mutant. Furthermore, a growth defect in copper-containing medium demonstrated that CopB functions as an export ATPase for copper ions. Furthermore, EMSAs revealed that CopR functions as a regulator that negatively regulates copB gene and Cu2+ serves as inducer of CopR to activate the expression of CopB in response to H2O2 stress in L. plantarum CAUH2. Our findings indicated that CopR plays an important role in enhancing oxidative resistance by regulating copB to modulate copper homeostasis. PMID:29089937

  5. Low-Lying Energy Isomers and Global Minima of Aqueous Nanoclusters: Structures and Spectroscopic Features of the Pentagonal Dodecahedron (H2O)20 and (H3O)+(H2O)20

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Xantheas, Sotiris S.

    We rely on a hierarchy of methods to identify the low-lying isomers for the pentagonal dodecahedron (H2O)20 and the H3O+(H2O)20 clusters. Initial screening of isomers was performed with classical potentials [TIP4P, TTM2-F, TTM2.1-F for (H2O)20 and ASP for H3O+(H2O)20] and the networks obtained with those potentials were subsequently reoptimized at the DFT (B3LYP) and MP2 levels of theory. For the pentagonal dodecahedron (H2O)20 it was found that DFT (B3LYP) and MP2 produced the same global minimum. However, this was not the case for the H3O+(H2O)20 cluster, for which MP2 produced a different network for the global minimum when compared tomore » DFT (B3LYP). All low-lying minima of H3O+(H2O)20 correspond to hydrogen bonding networks having 9 ''free'' OH bonds and the hydronium ion on the surface of the cluster. The fact that DFT (B3LYP) and MP2 produce different results and issues related to the use of a smaller basis set, explains the discrepancy between the current results and the structure previously suggested [Science 304, 1137 (2004)] for the global minimum of the H3O+(H2O)20 cluster. Additionally, the IR spectra of the MP2 global minimum are closer to the experimentally measured ones than the spectra of the previously suggested DFT global minimum. The latter exhibit additional bands in the most red-shifted region of the OH stretching vibrations (corresponding to the ''fingerprint'' of the underlying hydrogen bonding network), which are absent from both the experimental as well as the spectra of the new structure suggested for the global minimum of this cluster.« less

  6. Characterization of an intracellular oligopeptidase from Lactobacillus paracasei.

    PubMed Central

    Tobiassen, R O; Sørhaug, T; Stepaniak, L

    1997-01-01

    An intracellular oligopeptidase from Lactobacillus paracasei Lc-01 has been purified to homogeneity by Fast Flow Q Sepharose, hydroxyapatite, and Mono Q chromatography. The molecular mass of the enzyme was determined to be 140 kDa by gel filtration and approximately 30 kDa by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis and SDS-capillary electrophoresis. The pI of the enzyme was at pH 4.5. The enzyme expressed maximum activity at pH 8.0 and 40 degrees C. Oligopeptidase activity on bradykinin was inhibited strongly by 1,10-phenantroline and EDTA and partly by p-chloromercuribenzoic acid but not by phosphoramidon or phenylmethylsulfonyl fluoride. Marked inhibition by beta-casein fragment 58 to 72 was demonstrated. The enzyme showed neither general aminopeptidase nor caseinolytic activity, and it degraded only oligopeptides between 8 and 13 amino acids. The enzyme readily hydrolyzed the Phe-Ser and Pro-Phe bonds of bradykinin; the Phe-His bond of angiotensin I; the Pro-Gln, Gln-Phe, and Phe-Gly bonds of substance P; and the Pro-Tyr bond of neurotensin. Weak activity toward the Ala-Tyr and Pro-Ser bonds of alpha(s1)-casein fragment 157 to 164, was observed. The N-terminal amino acid sequence of the oligopeptidase showed a high degree of homology to the lactacin B inducer from Lactobacillus acidophilus. PMID:9097425

  7. Conversion of Thin Stillage Compounds using Endemic Bacteria Augmented with Lactobacillus panis PM1B.

    PubMed

    Ratanapariyanuch, Kornsulee; Shim, Youn Young; Reaney, Martin J T

    2016-10-04

    A consortium of organisms endemic in wheat-based thin stillage (W-TS) obtained from a commercial ethanol production converts glycerol to 1,3-propanediol (1,3-PD) and lactic acid to acetic acid. We sought to improve conditions for 1,3-PD and acetic acid production to be used in future studies of industrial isolation of these compounds from two-stage fermentation. Occasionally stillage fermentation proceeded slowly but an inoculum of Lactobacillus panis PM1B augmented both fermentation rate and extent. Fermentation rate and product yield were enhanced by adjusting pH to 5 daily, adding glucose and glycerol (molar ratio 0.1:1), adding freeze-dried W-TS, and adding vitamins (B 2 , B 3 , and B 12 ). 1,3-PD and 3-HPA did not inhibit 1,3-PD production during fermentation. Moreover, agitation did not improve fermentation rate or extent. Corn sugar was a suitable substitute for glucose. Fermentation was performed at both 20 and 150 L, with 1,3-PD production of 2% (w/v, 20 g/L) being routinely achieved or exceeded.

  8. Effect of yogurt containing polydextrose, Lactobacillus acidophilus NCFM and Bifidobacterium lactis HN019: a randomized, double-blind, controlled study in chronic constipation.

    PubMed

    Magro, Daniéla Oliveira; de Oliveira, Lais Mariana R; Bernasconi, Isabela; Ruela, Marilia de Souza; Credidio, Laura; Barcelos, Irene K; Leal, Raquel F; Ayrizono, Maria de Lourdes Stesuko; Fagundes, João José; Teixeira, Leandro de B; Ouwehand, Arthur C; Coy, Claudio S R

    2014-07-24

    Constipation is a frequent complaint and the combination of a prebiotic and probiotics could have a potentially synergic effect on the intestinal transit. The present study therefore aims to investigate the combination of polydextrose (Litesse), L. acidophilus NCFM® and B. lactis HN019 in a yogurt on intestinal transit in subjects who suffer from constipation. Patients with constipation were randomly divided into two groups, Control Group (CG) and Treatment Group (TG), and had to eat 180 ml of unflavored yogurt every morning for 14 days. Those in the CG received only yogurt, while the TG received yogurt containing polydextrose, L. acidophilus NCFM (ATCC 700396) and B. lactis HN019 (AGAL NM97/09513). Favourable clinical response was assessed since Agachan score had a significant reduction at the end of the study in both groups and tended to be better in the TG. The subjects in the treatment group also had a shorter transit time at the end of the intervention compared to the control group (p = 0.01). The product containing yogurt with polydextrose, B. lactis HN019 and L. acidophilus NCFM® significantly shortened colonic transit time after two weeks in the TG compared to CG and may be an option for treatment of constipation.

  9. The S-layer Associated Serine Protease Homolog PrtX Impacts Cell Surface-Mediated Microbe-Host Interactions of Lactobacillus acidophilus NCFM

    PubMed Central

    Johnson, Brant R.; O’Flaherty, Sarah; Goh, Yong Jun; Carroll, Ian; Barrangou, Rodolphe; Klaenhammer, Todd R.

    2017-01-01

    Health-promoting aspects attributed to probiotic microorganisms, including adhesion to intestinal epithelia and modulation of the host mucosal immune system, are mediated by proteins found on the bacterial cell surface. Notably, certain probiotic and commensal bacteria contain a surface (S-) layer as the outermost stratum of the cell wall. S-layers are non-covalently bound semi-porous, crystalline arrays of self-assembling, proteinaceous subunits called S-layer proteins (SLPs). Recent evidence has shown that multiple proteins are non-covalently co-localized within the S-layer, designated S-layer associated proteins (SLAPs). In Lactobacillus acidophilus NCFM, SLP and SLAPs have been implicated in both mucosal immunomodulation and adhesion to the host intestinal epithelium. In this study, a S-layer associated serine protease homolog, PrtX (prtX, lba1578), was deleted from the chromosome of L. acidophilus NCFM. Compared to the parent strain, the PrtX-deficient strain (ΔprtX) demonstrated increased autoaggregation, an altered cellular morphology, and pleiotropic increases in adhesion to mucin and fibronectin, in vitro. Furthermore, ΔprtX demonstrated increased in vitro immune stimulation of IL-6, IL-12, and IL-10 compared to wild-type, when exposed to mouse dendritic cells. Finally, in vivo colonization of germ-free mice with ΔprtX led to an increase in epithelial barrier integrity. The absence of PrtX within the exoproteome of a ΔprtX strain caused morphological changes, resulting in a pleiotropic increase of the organisms’ immunomodulatory properties and interactions with some intestinal epithelial cell components. PMID:28713337

  10. The Efficacy of Bifidobacterium longum BORI and Lactobacillus acidophilus AD031 Probiotic Treatment in Infants with Rotavirus Infection.

    PubMed

    Park, Myeong Soo; Kwon, Bin; Ku, Seockmo; Ji, Geun Eog

    2017-08-16

    A total of 57 infants hospitalized with rotavirus disease were included in this study. The children were randomly divided into the study's two treatment groups: three days of the oral administration of (i) a probiotics formula containing both Bifidobacterium longum BORI and Lactobacillus acidophilus AD031 ( N = 28); or (ii) a placebo (probiotic-free skim milk, N = 29) and the standard therapy for diarrhea. There were no differences in age, sex, or blood characteristics between the two groups. When the 57 cases completed the protocol, the duration of the patients' diarrhea was significantly shorter in the probiotics group (4.38 ± 1.29, N = 28) than the placebo group (5.61 ± 1.23, N = 29), with a p -value of 0.001. Symptoms such as duration of fever ( p = 0.119), frequency of diarrhea ( p = 0.119), and frequency of vomiting ( p = 0.331) tended to be ameliorated by the probiotic treatment; however, differences were not statistically significant between the two groups. There were no serious, adverse events and no differences in the frequency of adverse events in both groups.

  11. Probiotic bacteria Lactobacillus acidophilus NCFM and Bifidobacterium lactis Bi-07 versus placebo for the symptoms of bloating in patients with functional bowel disorders: a double-blind study.

    PubMed

    Ringel-Kulka, Tamar; Palsson, Olafur S; Maier, Danielle; Carroll, Ian; Galanko, Joseph A; Leyer, Gregory; Ringel, Yehuda

    2011-07-01

    Recent data suggest a role for the intestinal microbiota in the pathogenesis of functional bowel disorders (FBDs). Probiotic studies in FBDs generated inconsistent results suggesting a strain-specific and product-specific effect. To investigate the clinical efficacy of Lactobacillus acidophilus NCFM (L-NCFM) and Bifidobacterium lactis Bi-07 (B-LBi07) in nonconstipation FBDs. A double-blind, placebo-control clinical trial of the probiotic bacterias L-NCFM and B-LBi07 twice a day (2×10(11) CFU/d) versus placebo over 8 weeks. Primary endpoints were global relief of gastrointestinal symptoms and satisfaction with treatment. Secondary endpoints were change in symptoms severity, well-being, and quality of life. Microbiological effect was assessed by quantitative real time polymerase chain reaction on fecal samples. Sixty patients (probiotic, n=31; placebo, n=29), 72% females, 84% whites, mean age 37 years. Abdominal bloating improved in the probiotics compared with the placebo group at 4 weeks (4.10 vs 6.17, P=0.009; change in bloating severity P=0.02) and 8 weeks (4.26 vs 5.84, P=0.06; change in bloating severity P<0.01). Analyses on the irritable bowel syndrome subgroup (n=33) showed similar results. L-NCFM and B-LBi07 twice a day improve symptoms of bloating in patients with FBDs. These data supports the role of intestinal bacteria in the pathophysiology of FBD and the role for probiotic bacteria in the management of these disorders.

  12. Probiotic Dahi containing Lactobacillus acidophilus and Bifidobacterium bifidum alleviates age-inflicted oxidative stress and improves expression of biomarkers of ageing in mice.

    PubMed

    Kaushal, Deepti; Kansal, Vinod K

    2012-02-01

    The potential benefiting effects of probiotic Dahi on age-inflicted accumulation of oxidation products, antioxidant enzymes and expression of biomarkers of ageing were evaluated in mice. Probiotic Dahi were prepared by co-culturing in buffalo milk (3% fat) Dahi bacteria (Lactococcus lactis ssp. cremoris NCDC-86 and Lactococcus lactis ssp. lactis biovar diacetylactis NCDC-60) along with selected strain of Lactobacillus acidophilus LaVK2 (La-Dahi) or combined L. acidophilus and Bifidobacterium bifidum BbVK3 (LaBb-Dahi). Four groups of 12 months old mice (6 each) were fed for 4 months supplements (5 g/day) of buffalo milk (3% fat), Dahi, La-Dahi and LaBb-Dahi, respectively, with basal diet. The activities of catalase (CAT) and glutathione peroxidase (GPx) declined and the contents of oxidation products, thiobarbituric acid reactive substances (TBARS) and protein carbonyls, increased in red blood corpuscles (RBCs), liver, kidney and heart tissues and superoxide dismutase (SOD) activity increased in RBCs and hepatic tissues during ageing of mice. Feeding ageing mice with La-Dahi or LaBb-Dahi increased CAT activity in all the four tissues, and GPx activity in RBCs and hepatic tissue, and a significant decline in TBARS in plasma, kidney and hepatic tissues and protein carbonyls in plasma. Feeding mice with probiotic Dahi also reversed age related decline in expression of biomarkers of ageing, peroxisome proliferators activated receptor-α, senescence marker protein-30 (SMP-30) and klotho in hepatic and kidney tissues. The present study suggests that probiotic Dahi containing selected strains of bacteria can be used as a potential nutraceutical intervention to combat oxidative stress and molecular alterations associated with ageing.

  13. Anti-influenza virus effects of both live and non-live Lactobacillus acidophilus L-92 accompanied by the activation of innate immunity.

    PubMed

    Goto, Hiroaki; Sagitani, Atsuhiro; Ashida, Nobuhisa; Kato, Shinji; Hirota, Tatsuhiko; Shinoda, Tadashi; Yamamoto, Naoyuki

    2013-11-01

    The antiviral effects of both a live and non-live Lactobacillus acidophilus strain L-92 (L-92) were investigated by oral administration (10 mg/mouse per d) daily for 21 d in a mouse model infected intranasally with influenza virus (H1N1). Virus titres in the lung of mice administered either live or non-live L-92 cells daily for 15 d were repressed 6 d after virus infection compared with the control group. Natural killer (NK) activity in the orally administered non-live L-92 group was higher compared with that of the control group before virus infection and on day 6. In contrast, NK activity in the live L-92 group compared with the control group was not significantly changed on both days, but was significantly higher on day 1. In contrast, live L-92 showed a greater repression of virus proliferation compared with non-live L-92, 6 d after the infection. Live L-92 decreased the number of neutrophils in the lung and suppressed lung weight, leading to the consequent deterioration of consolidation scores of the lung. These results indicated that pretreatment of live or non-live L-92 cells had protective effects against influenza virus infection. Among the measured cytokines and chemokines, eotaxin, macrophage colony-stimulating factor, IL-1b, RANTES (regulated on activation, normal T cell expressed and secreted) and interferon-a were significantly increased in the lung: IL-17 was significantly increased in Peyer’s patch of the live L-92 group compared with the control group. A mechanistic study suggested that the enhancement of NK activity in the lung caused by stimulating various antiviral cytokines and chemokines after the oral administration of L-92 cells might be important in protecting against virus infection.

  14. Identification, characterization, immobilization, and mutational analysis of a novel acetylesterase with industrial potential (LaAcE) from Lactobacillus acidophilus.

    PubMed

    Wang, Ying; Ryu, Bum Han; Yoo, Wanki; Lee, Chang Woo; Kim, Kyeong Kyu; Lee, Jun Hyuck; Kim, T Doohun

    2018-01-01

    Lactic acid bacteria, which are involved in the fermentation of vegetables, meats, and dairy products, are widely used for the productions of small organic molecules and bioactive peptides. Here, a novel acetylesterase (LaAcE) from Lactobacillus acidophilus NCFM was identified, functionally characterized, immobilized, and subjected to site-directed mutagenesis for biotechnological applications. The enzymatic properties of LaAcE were investigated using biochemical and biophysical methods including native polyacrylamide gel electrophoresis, acetic acid release, biochemical assays, enzyme kinetics, and spectroscopic methods. Interestingly, LaAcE exhibited the ability to act on a broad range of substrates including glucose pentaacetate, glyceryl tributyrate, fish oil, and fermentation-related compounds. Furthermore, immobilization of LaAcE showed good recycling ability and high thermal stability compared with free LaAcE. A structural model of LaAcE was used to guide mutational analysis of hydrophobic substrate-binding region, which was composed of Leu 156 , Phe 164 , and Val 204 . Five mutants (L156A, F164A, V204A, L156A/F164A, and L156A/V204A) were generated and investigated to elucidate the roles of these hydrophobic residues in substrate specificity. This work provided valuable insights into the properties of LaAcE, and demonstrated that LaAcE could be used as a model enzyme of acetylesterase in lactic acid bacteria, making LaAcE a great candidate for industrial applications. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. prtH2, Not prtH, Is the Ubiquitous Cell Wall Proteinase Gene in Lactobacillus helveticus▿

    PubMed Central

    Genay, M.; Sadat, L.; Gagnaire, V.; Lortal, S.

    2009-01-01

    Lactobacillus helveticus strains possess an efficient proteolytic system that releases peptides which are essential for lactobacillus growth in various fermented dairy products and also affect textural properties or biological activities. Cell envelope proteinases (CEPs) are bacterial enzymes that hydrolyze milk proteins. In the case of L. helveticus, two CEPs with low percentages of amino acid identity have been described, i.e., PrtH and PrtH2. However, the distribution of the genes that encode CEPs still remains unclear, rendering it difficult to further control the formation of particular peptides. This study evaluated the diversity of genes that encode CEPs in a collection of strains of L. helveticus isolated from various biotopes, both in terms of the presence or absence of these genes and in terms of nucleotide sequence, and studied their transcription in dairy matrices. After defining three sets of primers for both the prtH and prtH2 genes, we studied the distribution of the genes by using PCR and Southern blotting experiments. The prtH2 gene was ubiquitous in the 29 strains of L. helveticus studied, whereas only 18 of them also exhibited the prtH gene. Sequencing of a 350-bp internal fragment of these genes revealed the existence of intraspecific diversity. Finally, expression of these two CEP-encoding genes was followed during the growth in dairy matrices of two strains, ITG LH77 and CNRZ32, which possess one and two CEP-encoding genes, respectively. Both genes were shown to be expressed by L. helveticus at each stage of growth in milk and at different stages of mini-Swiss-type cheese making and ripening. PMID:19286786

  16. Effect of lactobacillus strains on phenolic profile, color attributes and antioxidant activities of lactic-acid-fermented mulberry juice.

    PubMed

    Kwaw, Emmanuel; Ma, Yongkun; Tchabo, William; Apaliya, Maurice Tibiru; Wu, Meng; Sackey, Augustina Sackle; Xiao, Lulu; Tahir, Haroon Elrasheid

    2018-06-01

    This study was conducted to investigate the effect of lactic acid bacteria (LAB) strains on color properties, phenolic profile and antioxidant activities of mulberry juice. Mulberry juice was separately fermented at 37 °C for 36 h using Lactobacillus plantarum, Lactobacillus acidophilus and Lactobacillus paracasei. The results showed that lactic acid fermentation impacted on the color of the juice. Moreover, the study demonstrated that LABs impacted on the phenolic profile of the juice. Syringic acid, cyanidin-3-O-rutinoside and quercetin were the predominant phenolic acid, anthocyanin and flavonol respectively in the lactic-acid-fermented mulberry juice. The degree of radical scavenging activity was species-specific with the L. plantarum fermented juice having the highest radical scavenging activities. The correlation analysis demonstrated that flavonols and anthocyanins were mostly responsible for the increased in 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) scavenging activity while phenolic acids and flavonols were responsible for 2,2-diphenyl-1-picrylhydrazyl scavenging activity and reducing power capacity of the fermented juice. Copyright © 2018 Elsevier Ltd. All rights reserved.

  17. 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 μMT mice or Jh mice, and Lactobacillus-primed μMT mice had no characteristic infiltrates or airway Igs. Nonetheless, we observed diminished virus recovery and profound suppression of virus-induced proinflammatory cytokines CCL2, IFN-γ, and CXCL10 in both wild-type and Lactobacillus-primed μMT mice. Furthermore, Lactobacillus plantarum-primed, B cell-deficient μMT 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.

  18. Ultra-low-dose estriol and Lactobacillus acidophilus vaginal tablets (Gynoflor(®)) for vaginal atrophy in postmenopausal breast cancer patients on aromatase inhibitors: pharmacokinetic, safety, and efficacy phase I clinical study.

    PubMed

    Donders, Gilbert; Neven, Patrick; Moegele, Maximilian; Lintermans, Anneleen; Bellen, Gert; Prasauskas, Valdas; Grob, Philipp; Ortmann, Olaf; Buchholz, Stefan

    2014-06-01

    Phase I pharmacokinetic (PK) study assessed circulating estrogens in breast cancer (BC) patients on a non-steroidal aromatase inhibitor (NSAI) with vaginal atrophy using vaginal ultra-low-dose 0.03 mg estriol (E3) and Lactobacillus combination vaginal tablets (Gynoflor(®)). 16 women on NSAI with severe vaginal atrophy applied a daily vaginal tablet of Gynoflor(®) for 28 days followed by a maintenance therapy of 3 tablets weekly for 8 weeks. Primary outcomes were serum concentrations and PK of E3, estradiol (E2), and estrone (E1) using highly sensitive gas chromatography-mass spectrometry. Secondary outcomes were clinical measures for efficacy and side effects; microscopic changes in vaginal epithelium and microflora; and changes in serum FSH, LH, and sex hormone-binding globulin. Compared with baseline, serum E1 and E2 did not increase in any of the women at any time following vaginal application. Serum E3 transiently increased after the first application in 15 of 16 women, with a maximum of 168 pg/ml 2-3 h post-insertion. After 4 weeks, serum E3 was slightly increased in 8 women with a maximum of 44 pg/ml. The vaginal atrophy resolved or improved in all women. The product was well tolerated, and discontinuation of therapy was not observed. The low-dose 0.03 mg E3 and Lactobacillus acidophilus vaginal tablets application in postmenopausal BC patients during AI treatment suffering from vaginal atrophy lead to small and transient increases in serum E3, but not E1 or E2, and therefore can be considered as safe and efficacious for treatment of atrophic vaginitis in BC patients taking NSAIs.

  19. Comparative Evaluation of Antibacterial Effect of Dental Luting Cements on Streptococcus mutans and Lactobacillus acidophilus: An In vitro Study.

    PubMed

    Feroz, Sma; Bhoyar, A; Khan, S

    2016-12-01

    This study aimed at evaluating and comparing the antibacterial activity of six types of dental luting cements on Streptococcus mutans and Lactobacillus acidophilus using the agar diffusion test (ADT) and the direct-contact test (DCT). The antibacterial activity in ADT was measured based on the diameter of the zone of inhibition formed, whereas in DCT the density of the bacterial suspension was measured. The lower the density of the suspension, the more antibacterial activity the cement possesses. Agar diffusion test was carried out on the bacteria. After an incubation period of 24 hours, the plates were checked for the presence of zone of inhibition. In DCT the cement was mixed and applied. Once the cement was set, bacterial suspension and brain-heart infusion medium was poured and incubated for 24 hours. After 24 hours, the plate was placed in the enzyme-linked immunosorbent assay plate reader, which measured the optical density of the fluid. The first set of data was recorded approximately 1 hour after incubation. Overall, three sets of data were recorded. Additional experiments were performed on set test materials that were allowed to age for 24 hours, 1 week, 1, 3, and 6 months. When using ADT only two cements zinc oxide eugenol (ZOE) and zinc polycarboxylate (ZPC) cement showed antibacterial activity against the test organisms. When using DCT, all cements showed some amount of antibacterial activity. Zinc oxide eugenol and ZPC cement showed highest amount of antibacterial activity against S. mutans and L. acidophilus respectively. Within the limitations of study, ZOE cement and ZPC cement were most effective against the tested microorganisms followed by the newer resin cement. The glass ionomer cement was the weakest of all. Patients with high caries index can be treated more effectively using the abovementioned cements.

  20. Clinical effects of Lactobacillus acidophilus strain L-92 on perennial allergic rhinitis: a double-blind, placebo-controlled study.

    PubMed

    Ishida, Y; Nakamura, F; Kanzato, H; Sawada, D; Hirata, H; Nishimura, A; Kajimoto, O; Fujiwara, S

    2005-02-01

    Studies in animals have suggested that lactic acid bacteria alleviate allergic diseases, however, little information is available on their clinical effect on allergy in humans. Thus, we examined the efficacy of orally administered Lactobacillus acidophilus strain L-92 (L-92) on perennial allergic rhinitis. In a randomized, double-blind, placebo-controlled clinical trial, 49 patients with perennial allergic rhinitis were randomized to receive either 100 mL of heat-treated fermented milk containing L-92 (n = 25) or acidified milk without lactic acid bacteria (placebo; n = 24) for 8 wk. The severity of symptoms was evaluated based on the changes in the scores of clinical symptoms. Oral administration of milk fermented with L-92 resulted in a statistically significant improvement of nasal symptom-medication scores. Ocular symptom-medication scores of patients in the L-92 intervention group tended to improve compared with those in the placebo group. In addition, clear decreases of the scores of swelling and color of the nasal mucosa were observed in the L-92 intervention group at 6 and 8 wk after the start of ingestion of fermented milk. There were no significant differences in serum antihouse dust mite immunoglobulin E levels nor in T helper type 1/T helper type 2 ratio between the 2 groups. These results suggest that oral administration of L-92 can alleviate the symptoms of perennial allergic rhinitis, however, statistically significant changes were not shown in blood parameters.

  1. 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 μMT mice or Jh mice, and Lactobacillus-primed μMT 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 μMT mice. Furthermore, L. plantarum-primed, B-cell deficient μMT 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

  2. The Efficacy of Bifidobacterium longum BORI and Lactobacillus acidophilus AD031 Probiotic Treatment in Infants with Rotavirus Infection

    PubMed Central

    Park, Myeong Soo; Kwon, Bin; Ku, Seockmo; Ji, Geun Eog

    2017-01-01

    A total of 57 infants hospitalized with rotavirus disease were included in this study. The children were randomly divided into the study’s two treatment groups: three days of the oral administration of (i) a probiotics formula containing both Bifidobacterium longum BORI and Lactobacillus acidophilus AD031 (N = 28); or (ii) a placebo (probiotic-free skim milk, N = 29) and the standard therapy for diarrhea. There were no differences in age, sex, or blood characteristics between the two groups. When the 57 cases completed the protocol, the duration of the patients’ diarrhea was significantly shorter in the probiotics group (4.38 ± 1.29, N = 28) than the placebo group (5.61 ± 1.23, N = 29), with a p-value of 0.001. Symptoms such as duration of fever (p = 0.119), frequency of diarrhea (p = 0.119), and frequency of vomiting (p = 0.331) tended to be ameliorated by the probiotic treatment; however, differences were not statistically significant between the two groups. There were no serious, adverse events and no differences in the frequency of adverse events in both groups. PMID:28813007

  3. Characterization of Feruloyl Esterases Produced by the Four Lactobacillus Species: L. amylovorus, L. acidophilus, L. farciminis and L. fermentum, Isolated from Ensiled Corn Stover.

    PubMed

    Xu, Zhenshang; He, Huiying; Zhang, Susu; Guo, Tingting; Kong, Jian

    2017-01-01

    Lactic acid bacteria (LAB) play important roles in silage fermentation, which depends on the production of sufficient organic acids to inhibit the growth of undesirable microorganisms. However, LAB are not able to degrade cellulose and hemicellulose. Bacteria and fibrolytic enzymes are usually used as inoculants to improve the silage quality and digestibility. In the present study, we isolated four Lactobacillus strains ( L. amylovorus CGMCC 11056, L. acidophilus CCTCC AB2010208, L. farciminis CCTCC AB2016237 and L. fermentum CCTCC AB2010204) with feruloyl esterase (FAE) activities from ensiled corn stover (CS) by a plate screening assay. The genes encoding FAEs were cloned and hetero-expressed in Escherichia coli . The optimal temperature and pH of these purified enzymes ranged from 45 to 50°C and from 7.0 to 8.0, respectively. They could hydrolyze hydroxycinnamoyl esters in a substrate-specific manner when methyl ferulate, methyl caffeate, methyl ρ-coumarate and methyl sinapinate were used as substrates. Moreover, these four FAEs were able to hydrolyze CS to release hydroxycinnamic acids. Furthermore, these strains could degrade hydroxycinnamic esters, and L. amylovorus CGMCC 11056 was the most efficient strain among these four isolates. These results provided a new target for the development of inoculants to improve silage quality and digestibility.

  4. Characterization of Feruloyl Esterases Produced by the Four Lactobacillus Species: L. amylovorus, L. acidophilus, L. farciminis and L. fermentum, Isolated from Ensiled Corn Stover

    PubMed Central

    Xu, Zhenshang; He, Huiying; Zhang, Susu; Guo, Tingting; Kong, Jian

    2017-01-01

    Lactic acid bacteria (LAB) play important roles in silage fermentation, which depends on the production of sufficient organic acids to inhibit the growth of undesirable microorganisms. However, LAB are not able to degrade cellulose and hemicellulose. Bacteria and fibrolytic enzymes are usually used as inoculants to improve the silage quality and digestibility. In the present study, we isolated four Lactobacillus strains (L. amylovorus CGMCC 11056, L. acidophilus CCTCC AB2010208, L. farciminis CCTCC AB2016237 and L. fermentum CCTCC AB2010204) with feruloyl esterase (FAE) activities from ensiled corn stover (CS) by a plate screening assay. The genes encoding FAEs were cloned and hetero-expressed in Escherichia coli. The optimal temperature and pH of these purified enzymes ranged from 45 to 50°C and from 7.0 to 8.0, respectively. They could hydrolyze hydroxycinnamoyl esters in a substrate-specific manner when methyl ferulate, methyl caffeate, methyl ρ-coumarate and methyl sinapinate were used as substrates. Moreover, these four FAEs were able to hydrolyze CS to release hydroxycinnamic acids. Furthermore, these strains could degrade hydroxycinnamic esters, and L. amylovorus CGMCC 11056 was the most efficient strain among these four isolates. These results provided a new target for the development of inoculants to improve silage quality and digestibility. PMID:28626449

  5. A food additive with prebiotic properties of an α-d-glucan from lactobacillus plantarum DM5.

    PubMed

    Das, Deeplina; Baruah, Rwivoo; Goyal, Arun

    2014-08-01

    An α-d-glucan produced by Lactobacillus plantarum DM5 was explored for in vitro prebiotic activities. Glucan-DM5 demonstrated 21.6% solubility, 316.9% water holding capacity, 86.2% flocculation activity, 71.4% emulsification activity and a degradation temperature (Td) of 292.2°C. Glucan-DM5 exhibited lowest digestibility of 0.54% by artificial gastric juice, 0.21% by intestinal fluid and 0.32% by α-amylase whereas the standard prebiotic inulin, showed 25.23%, 5.97% and 19.13%, hydrolysis, respectively. Prebiotic activity assay of glucan-DM5 displayed increased growth of probiotic bacteria such as Bifidobacterium infantis and Lactobacillus acidophilus, but did not support the growth of non-probiotic bacteria such as Escherichia coli and Enterobacter aerogenes. The overall findings indicated that glucan from L. plantarum DM5 can serve as a potential prebiotic additive for food products. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. Isolation of a thermostable acid phytase from Aspergillus niger UFV-1 with strong proteolysis resistance

    PubMed Central

    Monteiro, Paulo S.; Guimarães, Valéria M.; de Melo, Ricardo R.; de Rezende, Sebastião T.

    2015-01-01

    An Aspergillus niger UFV-1 phytase was characterized and made available for industrial application. The enzyme was purified via ultrafiltration followed by acid precipitation, ion exchange and gel filtration chromatography. This protein exhibited a molecular mass of 161 kDa in gel filtration and 81 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), indicating that it may be a dimer. It presented an optimum temperature of 60 °C and optimum pH of 2.0. The K M for sodium phytate hydrolysis was 30.9 mM, while the k cat and k cat / K M were 1.46 ×10 5 s −1 and 4.7 × 10 6 s −1 .M −1 , respectively. The purified phytase exhibited broad specificity on a range of phosphorylated compounds, presenting activity on sodium phytate, p-NPP, 2- naphthylphosphate, 1- naphthylphosphate, ATP, phenyl-phosphate, glucose-6-phosphate, calcium phytate and other substrates. Enzymatic activity was slightly inhibited by Mg 2+ , Cd 2+ , K + and Ca 2+ , and it was drastically inhibited by F − . The enzyme displayed high thermostability, retaining more than 90% activity at 60 °C during 120 h and displayed a t 1/2 of 94.5 h and 6.2 h at 70 °C and 80 °C, respectively. The enzyme demonstrated strong resistance toward pepsin and trypsin, and it retained more than 90% residual activity for both enzymes after 1 h treatment. Additionally, the enzyme efficiently hydrolyzed phytate in livestock feed, liberating 15.3 μmol phosphate/mL after 2.5 h of treatment. PMID:26221114

  7. Strain-specific inhibition of the adherence of uropathogenic bacteria to bladder cells by probiotic Lactobacillus spp.

    PubMed

    de Llano, Dolores González; Arroyo, Amalia; Cárdenas, Nivia; Rodríguez, Juan Miguel; Moreno-Arribas, M Victoria; Bartolomé, Begoña

    2017-06-01

    Urinary tract infections (UTIs), one of most common infections worldwide, face high recurrence rates and increasing antimicrobial resistance. Probiotic bacteria, especially of the genus Lactobacillus, are considered a promising preventive and/or treatment therapy against UTIs. In order to elucidate the mechanisms involved in these beneficial effects, we studied the impact of different Lactobacillus strains (Lactobacillus salivarius UCM572, L. plantarum CLC17 and L. acidophilus 01) in the adherence of reference and clinical uropathogenic strains (Escherichia coli ATCC® 53503, E. coli 10791, Enterococcus faecalis 04-1, En. faecalis 08-1 and Staphylococcus epidermidis 08-3) to T24 epithelial bladder cells. In general, the Lactobacillus strains with previous in vivo evidence of beneficial effects against UTIs (L. salivarius UCM572 and L. acidophilus 01) significantly inhibited the adherence of the five uropathogens to T24 cells, displaying percentages of inhibition ranging between 22.2% and 43.9%, and between 16.5% and 53.7%, respectively. On the other hand, L. plantarum CLC17, a strain with no expected effects on UTIs, showed almost negligible anti-adherence effects.Therefore, these in vitro results suggest that inhibition of the adherence of uropathogens to epithelial bladder cells may be one of the mechanisms involved in the potential beneficial effects of probiotics against UTIs in vivo. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  8. Distribution Dynamics of Recombinant Lactobacillus in the Gastrointestinal Tract of Neonatal Rats

    PubMed Central

    Bao, Sujin; Zhu, Libin; Zhuang, Qiang; Wang, Lucia; Xu, Pin-Xian; Itoh, Keiji; Holzman, Ian R.; Lin, Jing

    2013-01-01

    One approach to deliver therapeutic agents, especially proteins, to the gastro-intestinal (GI) tract is to use commensal bacteria as a carrier. Genus Lactobacillus is an attractive candidate for use in this approach. However, a system for expressing exogenous proteins at a high level has been lacking in Lactobacillus. Moreover, it will be necessary to introduce the recombinant Lactobacillus into the GI tract, ideally by oral administration. Whether orally administered Lactobacillus can reach and reside in the GI tract has not been explored in neonates. In this study, we have examined these issues in neonatal rats. To achieve a high level of protein expression in Lactobacillus, we tested the impact of three promoters and two backbones on protein expression levels using mRFP1, a red fluorescent protein, as a reporter. We found that a combination of an L-lactate dehydrogenase (ldhL) promoter of Lactobacillus sakei with a backbone from pLEM415 yielded the highest level of reporter expression. When this construct was used to transform Lactobacillus casei, Lactobacillus delbrueckii and Lactobacillus acidophilus, high levels of mRFP1 were detected in all these species and colonies of transformed Lactobacillus appeared pink under visible light. To test whether orally administered Lactobacillus can be retained in the GI tract of neonates, we fed the recombinant Lactobacillus casei to neonatal rats. We found that about 3% of the bacteria were retained in the GI tract of the rats at 24 h after oral feeding with more recombinant Lactobacillus in the stomach and small intestine than in the cecum and colon. No mortality was observed throughout this study with Lactobacillus. In contrast, all neonatal rats died within 24 hours after fed with transformed E. coli. Taken together, our results indicate that Lactobacillus has the potential to be used as a vehicle for the delivery of therapeutic agents to neonates. PMID:23544119

  9. Statistical optimization of exopolysaccharide production by Lactobacillus plantarum NTMI05 and NTMI20.

    PubMed

    Imran, Mohamed Yousuff Mohamed; Reehana, Nazar; Jayaraj, K Arumugam; Ahamed, Abdul Azees Parveez; Dhanasekaran, Dharmadurai; Thajuddin, Nooruddin; Alharbi, Naiyf S; Muralitharan, Gangatharan

    2016-12-01

    In this study, 27 strains of Lactic acid bacteria (LAB) were isolated and identified from different milk sources. All the isolates were biochemically characterized and screened for their ability to produce exopolysaccharides (EPS), among which two isolates namely Lactobacillus plantarum NTMI05 (197mg/L) and Lactobacillus plantarum NTMI20 (187mg/L) showed higher EPS production. Both the isolates were molecular characterized and tested for their probiotic properties. The chemical composition of EPS from L. plantarum NTMI05 and NTMI20 revealed the presence of 95.45% and 92.35% carbohydrates, 14±0.1and 11±0.15mg/L lactic acid, 10.5±0.2 and 9±0.1mg/mL of reducing sugar, respectively. HPLC analysis showed galactose at the retention time of 2.29.The maximum EPS yield was optimized for the media components like glucose (20g/L), yeast extract (25g/L) and ammonium sulphate (2g/L) using Central Composite Design and Response Surface Methodology (RSM). Under optimum conditions the predicted maximum EPS production was 0.891g/L, 0.797g/L, while the actual experimental value was 0.956g/L and 0.827g/L for L. plantarum NTMI05 and NTMI20, respectively. The antioxidant capacity was also evaluated by DPPH and reducing power assay proving the potentiality of these organisms in food and dairy industries. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Effect of Fixed Orthodontic Treatment on Salivary Flow, pH and Microbial Count.

    PubMed

    Arab, Sepideh; Nouhzadeh Malekshah, Sepideh; Abouei Mehrizi, Ehsan; Ebrahimi Khanghah, Anita; Naseh, Roya; Imani, Mohammad Moslem

    2016-01-01

    The present study was designed to evaluate the changes in saliva properties and oral microbial flora in patients undergoing fixed orthodontic treatment. Two important saliva properties namely the salivary flow rate and pH as well as oral microbial flora were assessed in 30 orthodontic patients before starting fixed orthodontic treatment and after six, 12 and 18 weeks of treatment. Selective media, Sabouraud dextrose agar, Mitis salivarius agar and Rogosa agar were used for isolation of Candida albicans, Streptococcus mutans and Lactobacillus acidophilus, respectively. Statistical analysis was performed using Friedman and Dunn's tests. P< 0.05 was considered statistically significant. After six, 12 and 18 weeks of commencing fixed orthodontic treatment, the total colony counts of Candida albicans, Streptococcus mutans and Lactobacillus acidophilus showed a significant increase. The saliva pH decreased during the orthodontic treatment (P< 0.05) while the salivary flow did not change significantly. Fixed orthodontic treatment causes major changes in the saliva properties. The changes in oral microflora and saliva properties show the importance of caries preventive measures during orthodontic treatment.

  11. Complete genome sequence of Lactobacillus johnsonii FI9785, a competitive exclusion agent against pathogens in poultry.

    PubMed

    Wegmann, Udo; Overweg, Karin; Horn, Nikki; Goesmann, Alexander; Narbad, Arjan; Gasson, Michael J; Shearman, Claire

    2009-11-01

    Lactobacillus johnsonii is a member of the acidophilus group of lactobacilli. Because of their probiotic properties, including attachment to epithelial cells, immunomodulation, and competitive exclusion of pathogens, representatives of this group are being intensively studied. Here we report the complete annotated genome sequence of Lactobacillus johnsonii FI9785, a strain which prevents the colonization of specific-pathogen-free chicks by Clostridium perfringens.

  12. [Confocal laser scanning electron microscopy for assessment of vaginal Lactobacillus crispatus biofilm].

    PubMed

    Wu, Li-jie; Wang, Ben; Liao, Qin-ping; Zhang, Rui

    2015-12-18

    To investigate the female vaginal Lactobacillus crispatus biofilm by using confocal laser scanning microscopy (CLSM),thus revealing the formation of biofilm. The cover slide biofilm culture approach in vitro was employed for induction of the vaginal Lactobacillus crispatus biofilm formation. Following the culture for 2, 4, 8, 12, 16, 20, 24, 48, 72, 96 and 120 hours, the cover slide was removed for subsequent staining with the fluoresce in isothiocyanate-conjugated concanavalin A(FITC-ConA) and propidium (PI).This was followed by determination of the formation and characteristics of the vaginal Lactobacillus crispatus biofilm by using CLSM. The CLSM images of biofilm formation at different time points were captured, suggesting that the vaginal Lactobacillus crispatus adhesion occurred at h 4, which was in reversible attachment, then more and more Lactobacillus crispatus aggregated at h 8 to h 20, which was in irreversible attachment.Lactobacillus crispatus clustered at h 20, with early development of biofilm architecture.Then the biofilm with extracellular matrix around the bacteria was set up at h 24,with gradual matureation at h 24 to h 48.The biofilm dispersed at h 72. The biofilm density of cultivating for 20 hours was 42.7 × 10⁻³ ± 6.8 × 10⁻³ ,and for 24 hours increased to 102.5 × 10⁻³ ± 23.1 × 10⁻³, suggesting a significant difference, P<0.05. This meant that mature biofilm was formed at h 24. The vaginal Lactobacillus crispatus is able to form typical biofilm with distinct developmental phases and architecture characteristics.Mature biofilm is formed at h 24 to h 48, then the biofilm begins to disperse.

  13. Quantitative Real-Time PCR Analysis of Fecal Lactobacillus Species in Infants Receiving a Prebiotic Infant Formula

    PubMed Central

    Haarman, Monique; Knol, Jan

    2006-01-01

    The developing intestinal microbiota of breast-fed infants is considered to play an important role in the priming of the infants' mucosal and systemic immunity. Generally, Bifidobacterium and Lactobacillus predominate the microbiota of breast-fed infants. In intervention trials it has been shown that lactobacilli can exert beneficial effects on, for example, diarrhea and atopy. However, the Lactobacillus species distribution in breast-fed or formula-fed infants has not yet been determined in great detail. For accurate enumeration of different lactobacilli, duplex 5′ nuclease assays, targeted on rRNA intergenic spacer regions, were developed for Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus delbrueckii, Lactobacillus fermentum, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus reuteri, and Lactobacillus rhamnosus. The designed and validated assays were used to determine the amounts of different Lactobacillus species in fecal samples of infants receiving a standard formula (SF) or a standard formula supplemented with galacto- and fructo-oligosaccharides in a 9:1 ratio (OSF). A breast-fed group (BF) was studied in parallel as a reference. During the 6-week intervention period a significant increase was shown in total percentage of fecal lactobacilli in the BF group (0.8% ± 0.3% versus 4.1% ± 1.5%) and the OSF group (0.8% ± 0.3% versus 4.4% ± 1.4%). The Lactobacillus species distribution in the OSF group was comparable to breast-fed infants, with relatively high levels of L. acidophilus, L. paracasei, and L. casei. The SF-fed infants, on the other hand, contained more L. delbrueckii and less L. paracasei compared to breast-fed infants and OSF-fed infants. An infant milk formula containing a specific mixture of prebiotics is able to induce a microbiota that closely resembles the microbiota of BF infants. PMID:16597930

  14. Probiotic Properties of Lactobacillus Strains Isolated from Tibetan Kefir Grains

    PubMed Central

    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

  15. Functional and probiotic attributes of an indigenous isolate of Lactobacillus plantarum.

    PubMed

    Kaushik, Jai K; Kumar, Ashutosh; Duary, Raj K; Mohanty, Ashok K; Grover, Sunita; Batish, Virender K

    2009-12-01

    Probiotic microorganisms favorably alter the intestinal microflora balance, promote intestinal integrity and mobility, inhibit the growth of harmful bacteria and increase resistance to infection. Probiotics are increasingly used in nutraceuticals, functional foods or in microbial interference treatment. However, the effectiveness of probiotic organism is considered to be population-specific due to variation in gut microflora, food habits and specific host-microbial interactions. Most of the probiotic strains available in the market are of western or European origin, and a strong need for exploring new indigenous probiotic organisms is felt. An indigenous isolate Lp9 identified as Lactobacillus plantarum by molecular-typing methods was studied extensively for its functional and probiotic attributes, viz., acid and bile salt tolerance, cell surface hydrophobicity, autoaggregation and Caco-2 cell-binding as well as antibacterial and antioxidative activities. Lp9 isolate could survive 2 h incubation at pH 1.5-2.0 and toxicity of 1.5-2.0% oxgall bile. Lp9 could deconjugate major bile salts like glycocholate and deoxytaurocholate, indicating its potential to cause hypocholesterolemia. The isolate exhibited cell-surface hydrophobicity of approximately 37% and autoaggregation of approximately 31%. Presence of putative probiotic marker genes like mucus-binding protein (mub), fibronectin-binding protein (fbp) and bile salt hydrolase (bsh) were confirmed by PCR. Presence of these genes suggested the possibility of specific interaction and colonization potential of Lp9 isolate in the gut, which was also suggested by a good adhesion ratio of 7.4+/-1.3% with Caco-2 cell line. The isolate demonstrated higher free radical scavenging activity than standard probiotics L. johnsonii LA1 and L. acidophilus LA7. Lp9 also exhibited antibacterial activity against E. coli, L. monocytogenes, S. typhi, S. aureus and B. cereus. The indigenous Lactobacillus plantarum Lp9 exhibited high

  16. Techno-functional differentiation of two vitamin B12 producing Lactobacillus plantarum strains: an elucidation for diverse future use.

    PubMed

    Bhushan, Bharat; Tomar, S K; Chauhan, Arun

    2017-01-01

    An appropriate selection of Lactobacillus strain (probiotic/starter/functional) on the basis of its techno-functional characteristics is required before developing a novel fermented functional food. We compared vitamin B 12 (B 12 , cobalamin) producing Lactobacillus plantarum isolates, BHM10 and BCF20, for functional (vitamin over-production, genomic insight to B 12 structural genes, and probiotic attributes) and technological [milks (skim and soy) fermentation and B 12 bio-fortification] characteristics. Addition of B 12 precursors (5-amonolevulinate and dimethylbenzimidazole) to cobalamin-free fermentation medium increased vitamin production in BHM10, BCF20, and DSM20016 (a positive standard) by 3.4-, 4.4-, and 3.86-folds, respectively. Three important B 12 structural genes were detected in L. plantarum species (strains BHM10 and BCF20) by PCR for the first time. The gene sequences were submitted to NCBI GenBank and found phylogenetically closer to respective sequences in B 12 producing Lactobacillus reuteri strains. During comparative probiotic testing, BCF20 showed significantly higher (p < 0.05 to p < 0.001) gastrointestinal tolerance and cell surface hydrophobicity (p < 0.05) than BHM10. Moreover, only BCF20 was found positive for BSH activity and also exhibited comparatively better antagonistic potential against potent pathogens. Conversely, high acid and bile susceptible strain BHM10 displayed significantly higher soy milk fermentation and resultant B 12 bio-fortification abilities during technological testing. Two B 12 quantification techniques, UFLC and competitive immunoassay, confirmed the in vitro and in situ bio-production of bio-available form of B 12 after BHM10 fermentation. Conclusively, techno-functional differentiation of two B 12 producing strains elucidates their diverse future use; BCF20 either for B 12 over-production (in vitro) or as a probiotic candidate, while BHM10 for cobalamin bio-fortification (in situ) in soy milk.

  17. Effect of Non-Dairy Food Matrices on the Survival of Probiotic Bacteria during Storage

    PubMed Central

    Min, Min; Bunt, Craig R.; Mason, Susan L.; Bennett, Grant N.

    2017-01-01

    The viability of probiotics in non-dairy food products during storage is required to meet content criteria for probiotic products. This study investigated whether non-dairy foods could be matrices for probiotics. Selected probiotic bacteria were coated on non-dairy foods under two storage conditions, and viabilities were assessed. The non-dairy foods were coated with 5–7 log cfu g−1 of Lactobacillus acidophilus ATCC4356T, Lactobacillus plantarum RC30, and Bifidobacterium longum ATCC15707T. The coated non-dairy foods were stored at 20 °C and 20% relative humidity (RH) or 30 °C and 50% RH. Viability of probiotic bacteria was determined after 0, 2, and 4 weeks of storage. B. longum showed the highest survival at week 4 of 6.5–6.7 log cfu g−1 on wheat bran and oat, compared with 3.7–3.9 log cfu g−1 of L. acidophilus and 4.2–4.8 log cfu g−1 of L. plantarum at 20 °C 20% RH. Under the storage conditions of 30 °C 50% RH, survival of 4.5 log cfu g−1 of B. longum was also found on oat and peanut. This was two and four times higher than the population of L. acidophilus and L. plantarum, respectively. The results suggest that probiotics can survive on non-dairy foods under ambient storage conditions. However, the storage conditions, food matrices, and probiotic strains should be carefully chosen to maximize probiotic bacteria survival. PMID:28763015

  18. A gastrointestinal anti-infectious biotherapeutic agent: the heat-treated Lactobacillus LB

    PubMed Central

    Liévin-Le Moal, Vanessa

    2016-01-01

    Experimental in vitro and in vivo studies support the hypothesis that heat-treated, lyophilized Lactobacillus acidophilus LB cells and concentrated, neutralized spent culture medium conserve the variety of pharmacological, antimicrobial activities of the live probiotic strain against several infectious agents involved in well-established acute and persistent watery diarrhoea and gastritis. Heat-treated cells and heat-stable secreted molecules trigger multiple strain-specific activities explaining the therapeutic efficacy of L. acidophilus LB. This review discusses the current body of knowledge on the antimicrobial mechanisms of action exerted by L. acidophilus LB demonstrated in in vitro and in vivo experimental studies, and the evidence for the therapeutic efficacy of this anti-infectious biotherapeutic agent proved in randomized clinical trials for the treatment of acute and persistent watery diarrhoea associated with several intestinal infectious diseases in humans. PMID:26770268

  19. Importance of Molecular Methods to Determine Whether a Probiotic is the Source of Lactobacillus Bacteremia.

    PubMed

    Aroutcheva, Alla; Auclair, Julie; Frappier, Martin; Millette, Mathieu; Lolans, Karen; de Montigny, Danielle; Carrière, Serge; Sokalski, Stephen; Trick, William E; Weinstein, Robert A

    2016-03-01

    There has been an increasing interest in the use of probiotic products for the prevention of Clostridium difficile infection (CDI). Bio-K+(®) is a commercial probiotic product comprising three strains of lactobacilli--Lactobacillus acidophilus CL1285(®), Lact. casei LBC80R(®) and Lact. rhamnosus CLR2(®)--that have been applied to prevent CDI. Generally considered as safe, lactobacilli have potential to cause bacteremia, endocarditis and other infections. The source of Lactobacillus bacteremia can be normal human flora or lactobacilli-containing probiotic. The aim of this study was to assess whether probiotic lactobacilli caused bacteremia and to show the value of molecular identification and typing techniques to determine probiotic and patient strain relatedness. We report an episode of Lactobacillus bacteremia in a 69-year-old man admitted to a hospital with severe congestive heart failure. During his hospitalization, he required long-term antibiotic therapy. Additionally, the patient received Bio-K+(®) probiotic as part of a quality improvement project to prevent CDI. Subsequently, Lactobacillus bacteremia occurred. Two independent blinded laboratory evaluations, using pulse field gel electrophoresis, 16S rRNA gene sequencing and DNA fingerprint analysis (rep-PCR), were performed to determine whether the recovered Lact. acidophilus originated from the probiotic product. Ultimately, the patient strain was identified as Lact. casei and both laboratories found no genetic relation between the patient's strain and any of the probiotic lactobacilli. This clinical case of lactobacillus bacteremia in the setting of probiotic exposure demonstrates the value of using discriminatory molecular methods to clearly determine whether there were a link between the patient's isolate and the probiotic strains.

  20. Synbiotic Lactobacillus acidophilus NCFM and cellobiose does not affect human gut bacterial diversity but increases abundance of lactobacilli, bifidobacteria and branched-chain fatty acids: a randomized, double-blinded cross-over trial.

    PubMed

    van Zanten, Gabriella C; Krych, Lukasz; Röytiö, Henna; Forssten, Sofia; Lahtinen, Sampo J; Abu Al-Soud, Waleed; Sørensen, Søren; Svensson, Birte; Jespersen, Lene; Jakobsen, Mogens

    2014-10-01

    Probiotics, prebiotics, and combinations thereof, that is synbiotics, have been reported to modulate gut microbiota of humans. In this study, effects of a novel synbiotic on the composition and metabolic activity of human gut microbiota were investigated. Healthy volunteers (n = 18) were enrolled in a double-blinded, randomized, and placebo-controlled cross-over study and received synbiotic [Lactobacillus acidophilus NCFM (10(9) CFU) and cellobiose (5 g)] or placebo daily for 3 weeks. Fecal samples were collected and lactobacilli numbers were quantified by qPCR. Furthermore, 454 tag-encoded amplicon pyrosequencing was used to monitor the effect of synbiotic on the composition of the microbiota. The synbiotic increased levels of Lactobacillus spp. and relative abundances of the genera Bifidobacterium, Collinsella, and Eubacterium while the genus Dialister was decreased (P < 0.05). No other effects were found on microbiota composition. Remarkably, however, the synbiotic increased concentrations of branched-chain fatty acids, measured by gas chromatography, while short-chain fatty acids were not affected. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

  1. Antibacterial Effects of Different Concentrations of Althaea officinalis Root Extract versus 0.2% Chlorhexidine and Penicillin on Streptococcus mutans and Lactobacillus (In vitro).

    PubMed

    Haghgoo, Roza; Mehran, Majid; Afshari, Elahe; Zadeh, Hamide Farajian; Ahmadvand, Motahare

    2017-01-01

    The aims of the present study were to determine and compare the effects of different concentrations of Althaea officinalis extract, 0.2% chlorhexidine (CHX), and penicillin on Streptococcus mutans and Lactobacillus acidophilus in vitro . The laboratory study was done, for a period of 8 weeks. Minimum inhibitory concentration (MIC) in the test tube, minimum bactericidal concentration (MBC) in a plate culture medium, and growth inhibition zone diameter methods were used to compare the antibacterial effects of 0.2% CHX, penicillin, and different concentrations of A. officinalis root extract. The data were analyzed by SPSS version 24 using ANOVA and t -test analysis. The results showed A. officinalis root extract had antibacterial effect, but significant differences were in MIC and MBC against L. acidophilus and S. mutans with penicillin and 0.2% CHX mouthwash. In addition, the mean growth inhibition zones of all the concentrations of the plant extract were less than that of the positive control group ( P = 0.001). However, the difference in the maximum growth inhibition zone from that with the negative control group was significant. In addition, the antibacterial effect of the extract increased with an increase in its concentration. The extract exerted a greater antibacterial effect on S. mutans than on L. acidophilus . The plant polyphenols content is 3.7% which is equivalent to 29.93 g/ml. The root extract of A. officinalis exhibited antibacterial effects on S. mutans and L. acidophilus , but this effect was less than those of CHX mouthwash and penicillin. The antibacterial effect increased with an increase in the concentration of the extract.

  2. Characterization of biosurfactants produced by Lactobacillus spp. and their activity against oral streptococci biofilm.

    PubMed

    Ciandrini, Eleonora; Campana, Raffaella; Casettari, Luca; Perinelli, Diego R; Fagioli, Laura; Manti, Anita; Palmieri, Giovanni Filippo; Papa, Stefano; Baffone, Wally

    2016-08-01

    Lactic acid bacteria (LAB) can interfere with pathogens through different mechanisms; one is the production of biosurfactants, a group of surface-active molecules, which inhibit the growth of potential pathogens. In the present study, biosurfactants produced by Lactobacillus reuteri DSM 17938, Lactobacillus acidophilus DDS-1, Lactobacillus rhamnosus ATCC 53103, and Lactobacillus paracasei B21060 were dialyzed (1 and 6 kDa) and characterized in term of reduction of surface tension and emulsifying activity. Then, aliquots of the different dialyzed biosurfactants were added to Streptococcus mutans ATCC 25175 and Streptococcus oralis ATCC 9811 in the culture medium during the formation of biofilm on titanium surface and the efficacy was determined by agar plate count, biomass analyses, and flow cytometry. Dialyzed biosurfactants showed abilities to reduce surface tension and to emulsifying paraffin oil. Moreover, they significantly inhibited the adhesion and biofilm formation on titanium surface of S. mutans and S. oralis in a dose-dependent way, as demonstrated by the remarkable decrease of cfu/ml values and biomass production. The antimicrobial properties observed for dialyzed biosurfactants produced by the tested lactobacilli opens future prospects for their use against microorganisms responsible of oral diseases.

  3. Assessment of potential probiotic properties of Lactobacillus spp., Lactococcus spp., and Pediococcus spp. strains isolated from kefir.

    PubMed

    Sabir, Firat; Beyatli, Yavuz; Cokmus, Cumhur; Onal-Darilmaz, Derya

    2010-01-01

    In this study, the metabolic activities (in terms of quantities of the produced lactic acid, hydrogen peroxide, and exopolysaccharides) of 8 strains of Lactobacillus spp., Lactococcus spp., and Pediococcus spp., were determined. Lactic acid levels produced by strains were 8.1 to 17.4 mg/L. The L. acidophilus Z1L strain produced the maximum amount (3.18 μg/mL) of hydrogen peroxide. The exopolysaccharides (EPS) production by the strains was ranged between 173 and 378 mg/L. The susceptibility of 7 different antibiotics against these strains was also tested. All strains were found to be sensitive to ampicillin. The tolerance of the strains to low pH, their resistance to bile salts of strains, and their abilities to autoaggregate and coaggregate with Escherichia coli ATCC 11229 were also evaluated. High EPS-producing strains showed significant autoaggregation and coaggregation ability with test bacteria (P < 0.01). A correlation also was determined between EPS production and acid-bile tolerance (P < 0.05). EPS production possibly affects or is involved in acid-bile tolerance and aggregation of Lactobacillus spp., Lactococcus spp., and Pediococcus spp. strains and supports the potential of L. acidophilus Z1L strain as new probiotic. © 2010 Institute of Food Technologists®

  4. Three new insertion sequence elements ISLdl2, ISLdl3, and ISLdl4 in Lactobacillus delbrueckii: isolation, molecular characterization, and potential use for strain identification.

    PubMed

    Ravin, Victor; Alatossava, Tapani

    2003-05-01

    A group of new insertion sequence (IS) elements, ISLdl2, ISLdl3, and ISLdl4, from Lactobacillus delbrueckii subsp. lactis ATCC 15808 was isolated, characterized, and used for strain identification together with ISLdl1, recently characterized as an L. delbrueckii IS element belonging to the ISL3 family. ISLdl2 was 1367 bp in size and had a 24 bp IR and an 8 bp DR. The single ORF of ISLdl2 encoded a protein of 392 aa similar to transposases of the IS256 family. ISLdl3 had a single ORF encoding a protein of 343 aa similar to transposases of the IS30 family. Finally, ISLdl4 had a single ORF encoding a protein of 406 aa and displayed homology to the transposases of the IS110 family. ISLdl4 was only slight different from ISL4 (Accession No. AY040213). ISLdl1, ISLdl2, and ISLdl4 were present in all of the 10 L. delbrueckii subsp. lactis and subsp. delbrueckii strains tested, as well as in three of the 11 L. delbrueckii subsp. bulgaricus strains tested. ISLdl3 was present only in four closely related strains of L. delbrueckii subsp. lactis. These IS elements were not observed in Lactobacillus rhamnosus, Lactobacillus acidophilus, Lactobacillus helveticus, or Lactobacillus plantarum. A cluster of IS elements, ISLdl1, ISLdl2, ISLdl3, ISLdl4, and ISL6, was observed in L. delbrueckii subsp. lactis strain ATCC 15808. Within this cluster, ISLdl4 was inserted into ISLdl1 between the left IR and the start codon of ORF455, encoding a putative transposase. Most of the integration sites of the IS elements were strain-specific. We have observed that IS elements can migrate from one strain to another as integral parts of bacterial DNA by using phage LL-H as a vehicle. We demonstrate for the first time that inverse PCR and vectorette PCR methods with primers based on sequences of the IS elements could be used for identification of L. delbrueckii strains.

  5. Na{sub 3}[B{sub 20}H{sub 17}NH{sub 3}]: Synthesis and liposomal delivery to murine tumors

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Feakes, D.A.; Shelly, K.; Knobler, C.B.

    1994-04-12

    The polyhedral borane ion [n-B{sub 20}H{sub 18}]{sup 2{minus}} reacts with liquid ammonia in the presence of a suitable base to produce an apical-equatorial (ae) isomer of the [B{sub 20}H{sub 17}NH{sub 3}]{sup 3{minus}} ion, [1-(2{prime}-B{sub 10}H{sub 9})-2-NH{sub 3}B{sub 10}H{sub 8}]{sup 3{minus}}. The structure of this product has been confirmed by {sup 11}B NMR spectroscopy and x-ray crystallography. This species undergoes acid-catalyzed rearrangement to an apical-apical (a{sup 2}) isomer, [1-(1{prime}-B{sub 10}H{sub 9})-2-NH{sub 3}B{sub 10}H{sub 8}]{sup 3{minus}}, whose structure has been determined by {sup 11}B NMR spectroscopy. The sodium salts of both the ae and the a{sup 2} isomers of [B{sub 20}H{sub 17}NH{submore » 3}]{sup 3{minus}} have been encapsulated within small unilamellar liposomes, composed of distearoyl phosphatidyl-choline/cholesterol (1:1), and investigated as boron-delivery agents for boron neutron capture therapy (BNCT) of cancer. The biodistribution of boron was determined after the injection of liposomal suspensions into BALB/c mice bearing EMT6 tumors. Both [B{sub 20}H{sub 17}NH{sub 3}]{sup 3{minus}} isomers exhibited excellent tumor uptake and selectivity at very low injected doses, achieving peak tumor boron concentrations of 30-40 {mu}g of B/g of tissue and tumor/blood boron ratios of {approximately}5. The enhanced retention of the [B{sub 20}H{sub 17}NH{sub 3}]{sup 3{minus}} isomers by EMT6 tumors may be attributed to their facile intracellular oxidation. In another experiment, [ae-B{sub 20}H{sub 17}NH{sub 3}]{sup 3{minus}} was encapsulated in liposomes prepared with 5% PEG-2000-distearoyl phosphatidylethanolamine in the liposome membrane. As expected, these liposomes exhibited a longer circulation lifetime in the biodistribution experiment, resulting in the continued accumulation of boron in the tumor over the entire 48-hr experiment and reaching a maximum of 47 {mu}g of B/g of tumor.« less

  6. Targeting Mucosal Dendritic Cells with Microbial Antigens from Probiotic Lactic Acid Bacteria

    DTIC Science & Technology

    2008-03-01

    Lactoba- cillus gasseri, Lactobacillus plantarum , Lactobacillus delbreuckii, Lactobacillus rhamnosus, Lactobacillus salivarius and Lactobacillus ... Lactobacillus plantarum Helicobacter pylori UreB Mouse [105] S. pneumoniae PsaA Mouse [104] Lactococcus lactis C. tetani TTFC Mouse [81...anthracis (the causative agent of anthrax). An antigen-specific immune response can be elicited using specific strains of Lactobacillus acidophilus

  7. Effect of oral administration of Bacillus coagulans B37 and Bacillus pumilus B9 strains on fecal coliforms, Lactobacillus and Bacillus spp. in rat animal model

    PubMed Central

    Haldar, Lopamudra; Gandhi, D. N.

    2016-01-01

    Aim: To investigate the effect of oral administration of two Bacillus strains on fecal coliforms, Lactobacillus and Bacillus spp. in rat animal model. Materials and Methods: An in vivo experiment was conducted for 49-day period on 36 adult male albino Wister rats divided equally into to four groups. After 7-day adaptation period, one group (T1) was fed on sterile skim milk along with basal diet for the next 28 days. Second (T2) and (T3) groups received spore biomass of Bacillus coagulans B37 and Bacillus pumilus B9, respectively, suspended in sterilized skim milk at 8-9 log colony-forming units/ml plus basal diet for 28 days, while control group (T4) was supplied with clean water along with basal diet. There was a 14-day post-treatment period. A total of 288 fecal samples (8 fecal collections per rat) were collected at every 7-day interval starting from 0 to 49 days and subjected to the enumeration of the counts of coliforms and lactobacilli and Bacillus spores using respective agar media. In vitro acid and bile tolerance tests on both the strains were performed. Results: The rats those (T2 and T3) received either B. coagulans B37 or B. pumilus B9 spore along with non-fermented skim milk showed decrease (p<0.01) in fecal coliform counts and increase (p<0.05) in both fecal lactobacilli and Bacillus spore counts as compared to the control group (T4) and the group fed only skim milk (T1). In vitro study indicated that both the strains were found to survive at pH 2.0 and 3.0 even up to 3 h and tolerate bile up to 2.0% concentration even after 12 h of exposure. Conclusions: This study revealed that oral administration of either B. coagulans B37 or B. pumilus B9 strains might be useful in reducing coliform counts accompanied by concurrent increase in lactobacilli counts in the intestinal flora in rats. PMID:27536040

  8. Effect of oral administration of Bacillus coagulans B37 and Bacillus pumilus B9 strains on fecal coliforms, Lactobacillus and Bacillus spp. in rat animal model.

    PubMed

    Haldar, Lopamudra; Gandhi, D N

    2016-07-01

    To investigate the effect of oral administration of two Bacillus strains on fecal coliforms, Lactobacillus and Bacillus spp. in rat animal model. An in vivo experiment was conducted for 49-day period on 36 adult male albino Wister rats divided equally into to four groups. After 7-day adaptation period, one group (T1) was fed on sterile skim milk along with basal diet for the next 28 days. Second (T2) and (T3) groups received spore biomass of Bacillus coagulans B37 and Bacillus pumilus B9, respectively, suspended in sterilized skim milk at 8-9 log colony-forming units/ml plus basal diet for 28 days, while control group (T4) was supplied with clean water along with basal diet. There was a 14-day post-treatment period. A total of 288 fecal samples (8 fecal collections per rat) were collected at every 7-day interval starting from 0 to 49 days and subjected to the enumeration of the counts of coliforms and lactobacilli and Bacillus spores using respective agar media. In vitro acid and bile tolerance tests on both the strains were performed. The rats those (T2 and T3) received either B. coagulans B37 or B. pumilus B9 spore along with non-fermented skim milk showed decrease (p<0.01) in fecal coliform counts and increase (p<0.05) in both fecal lactobacilli and Bacillus spore counts as compared to the control group (T4) and the group fed only skim milk (T1). In vitro study indicated that both the strains were found to survive at pH 2.0 and 3.0 even up to 3 h and tolerate bile up to 2.0% concentration even after 12 h of exposure. This study revealed that oral administration of either B. coagulans B37 or B. pumilus B9 strains might be useful in reducing coliform counts accompanied by concurrent increase in lactobacilli counts in the intestinal flora in rats.

  9. Thermostability of xylanolytic enzymes produced by Lentinula edodes UFV70

    PubMed Central

    Ribeiro, Liliane Fraga Costa; Vaz, Marcelo Gomes Marçal Vieira; Chaves-Alves, Virgínia Maria; Vanetti, Maria Cristina Dantas; Vanetti, Maria Cristina Dantas; Kasuya, Maria Catarina Megumi; Passos, Flávia Maria Lopes; do Nascimento, Antônio Galvão

    2012-01-01

    Xylanolytic enzymes produced by Lentinula edodes UFV70, cultivated in eucalyptus sawdust/rice bran medium, were stable at 50, 60 and 65°C for 21 hours, losing only 15–25% activity. Fungus incubation at 50°C for 12 hours and at 65°C for 24 hours increased the amount of xylose produced. PMID:24031818

  10. A new record of Chironomus (Chironomus) acidophilus Keyl (Diptera, Chironomidae) from the Uzon volcanic caldera (Kronotsky Reserve, Kamchatka Peninsula, Russia), its karyotype, ecology and biology.

    PubMed

    Orel, Oksana V; Lobkova, Ludmila E; Zhirov, Sergey V; Petrova, Ninel A

    2015-07-03

    Morphology, cytology, ecology and biology of Holarctic Chironomus (Chironomus) acidophilus Keyl, 1960 (Diptera, Chironomidae) was examined from material collected in the geothermal Vosmerka Lake (pH=2.0-2.5). An illustrated redescription of C. acidophilus is given on the basis of adult males reared from field-collected pupae, and of simultaneously collected larvae. Additional larvae belonging to the pseudothummi-complex were identified as C. acidophilus on the basis of their karyotype. The karyotype of C. acidophilus (2n=8) and detailed mapping of the 4 chromosome arms A, E, D and F are provided. The population of C. acidophilus from Kamchatka was found to be karyologically monomorphic. Information on distribution and ecology of C. acidophilus from Vosmerka Lake (total mineralization 1583.5 mg/l) is also given. Chironomus acidophilus is the only species of aquatic insects recorded in this lake. Lack of competition and a richness of food resources contribute to the high abundance (35161 ind./m2) and biomass (11.342 g/m2) of the larvae of C. acidophilus in Vosmerka Lake.

  11. Short communication: Viability of culture organisms in honey-enriched acidophilus-bifidus-thermophilus (ABT)-type fermented camel milk.

    PubMed

    Varga, L; Süle, J; Nagy, P

    2014-11-01

    The aim of this research was to monitor the survival during refrigerated storage of Lactobacillus acidophilus LA-5 (A), Bifidobacterium animalis ssp. lactis BB-12 (B), and Streptococcus thermophilus CHCC 742/2130 (T) in cultured dairy foods made from camel and, for comparison, cow milks supplemented with black locust (Robinia pseudoacacia L.) honey and fermented by an acidophilus-bifidus-thermophilus (ABT)-type culture. Two liters of dromedary camel milk and 2 L of cow milk were heated to 90 °C and held for 10 min, then cooled to 40 °C. One half of both types of milk was fortified with black locust honey at the rate of 5.0% (wt/vol), whereas the other half was devoid of honey and served as a control. The camel and cow milks with and without honey were subsequently inoculated with ABT-5 culture and were fermented at 37 °C until a pH value of 4.6 was reached. Thereafter, the probiotic fermented milks were cooled to 15 °C in ice water and were each separated into 18 fractions that were transferred in sterile, tightly capped centrifuge tubes. After 24 h of cooling at 8 °C (d 0), the samples were stored at refrigeration temperature (4 °C). Three tubes of all 4 products (i.e., fermented camel and cow milks with and without honey) were taken at each sampling time (i.e., following 0, 7, 14, 21, 28, and 35 d of storage), and the counts of characteristic microorganisms and those of certain spoilage microbes (yeasts, molds, coliforms, Escherichia coli) were enumerated. The entire experimental program was repeated twice. The results showed that addition of black locust honey at 5% to heat-treated camel and cow milks did not influence the growth and survival of starter streptococci during production and subsequent refrigerated storage of fermented ABT milks. In contrast, honey improved retention of viability of B. animalis ssp. lactis BB-12 in the camel milk-based product during storage at 4 °C up to 5 wk. No spoilage organisms were detected in any of the samples tested

  12. [Clinical effects of Lactobacillus acidophilus strain L-55-contained yogurt on symptoms of Japanese cedar pollen allergy].

    PubMed

    Kimura, Goro; Akagi, Hirofumi; Okada, Chiharu; Hirano, Atsushi; Amano, Yoshimi; Ohmura, Etsuko; Nakashige, Yoshito; Sunada, Yosuke; Fujii, Yusuke; Nakamura, Syoji; Soda, Ryo; Takahashi, Kiyoshi

    2012-05-01

    It has been reported that oral administration of Lactobacillus acidophilus strain L-55 (L-55) suppressed nasal symptom and antigen-specific IgE induced by antigen challenge in mice. We investigated clinical effects of L-55-contained yogurt on symptoms and IgE production in the patients with Japanese cedar pollinosis. The study was performed as a randomized double blind placebo-controlled trial. L-55-contained yogurt (L-55 yogurt) or placebo yogurt was started to administer to each subject from 4 weeks prior to the onset of Japanese cedar pollen release for total 13 weeks. The mean value of symptom score and symptom medication score were lower in L-55 yogurt group compared with placebo yogurt group during 5th week to 9th week from the first week of Japanese cedar pollination. Especially, in medicated subjects, total symptom score and throat symptom score were significantly lower in L-55 yogurt group compared with placebo yogurt group at 5th and 4th week from the first week of Japanese cedar pollination, respectively. Moreover, in medicated subjects, change ratio of serum total IgE was significantly lower in L-55 yogurt group compared with placebo yogurt group at 1st week from the first week of Japanese cedar pollination. Intake of L-55 during Japanese cedar pollinating season may be effective in alleviating the allergic symptoms related to Japanese cedar pollinosis. From these finding, it was suggested that L-55 is a possible candidate as a complementary medicine for Japanese cedar pollinosis.

  13. Physiological functions at single-cell level of Lactobacillus spp. isolated from traditionally fermented cabbage in response to different pH conditions.

    PubMed

    Olszewska, Magdalena A; Kocot, Aleksandra M; Łaniewska-Trokenheim, Łucja

    2015-04-20

    Changes in pH are significant environmental stresses that may be encountered by lactobacilli during fermentation processes or passage through the gastrointestinal tract. Here, we report the cell response of Lactobacillus spp. isolated from traditionally fermented cabbage subjected to acid/alkaline treatments at pH 2.5, 7.4 and 8.1, which represented pH conditions of the gastrointestinal tract. Among six isolates, four species of Lactobacillus plantarum and two of Lactobacillus brevis were identified by fluorescence in situ hybridization (FISH). The fluorescence-based strategy of combining carboxyfluorescein diacetate (CFDA) and propidium iodine (PI) into a dual-staining assay was used together with epifluorescence microscopy (EFM) and flow cytometry (FCM) for viability assessment. The results showed that the cells maintained esterase activity and membrane integrity at pH 8.1 and 7.4. There was also no loss of culturability as shown by plate counts. In contrast, the majority of 2.5 pH-treated cells had a low extent of esterase activity, and experienced membrane perturbation. For these samples, an extensive loss of culturability was demonstrated. Comparison of the results of an in situ assessment with that of the conventional culturing method has revealed that although part of the stressed population was unable to grow on the growth media, it was deemed viable using a CFDA/PI assay. However, there was no significant change in the cell morphology among pH-treated lactobacilli populations. These analyses are expected to be useful in understanding the cell response of Lactobacillus strains to pH stress and may facilitate future investigation into functional and industrial aspects of this response. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Lactobacillus plantarum B7 inhibits Helicobacter pylori growth and attenuates gastric inflammation

    PubMed Central

    Sunanliganon, Chompoonut; Thong-Ngam, Duangporn; Tumwasorn, Somying; Klaikeaw, Naruemon

    2012-01-01

    AIM: To determine the anti-Helicobacter property of Lactobacillus plantarum B7 (L. plantarum) B7 supernatants in vitro and the protective effects of L. plantarum B7 on serum tumor necrosis factor-alpha (TNF-α), gastric malondialdehyde (MDA) level, apoptosis, and histopathology in Helicobacter pylori (H. pylori)-induced gastric inflammation in rats. METHODS: In vitro, the inhibition of H. pylori growth was examined using L. plantarum B7 supernatants at pH 4 and pH 7 and at the concentration of 1×, 5× and 10× on plates inoculated with H. pylori. The inhibitory effect of H. pylori was interpreted by the size of the inhibition zone. In vitro, male Sprague-Dawley rats were randomly divided into four groups including group 1 (control group), group 2 (H. pylori infected group), group 3 (H. pylori infected with L. plantarum B7 106 CFUs/mL treated group) and group 4 (H. pylori infected with L. plantarum B7 1010 CFUs/mL treated group). One week after H. pylori inoculation, L. plantarum B7 106 CFUs/mL or 1010 CFUs/mL were fed once daily to group 3 and group 4, respectively, for one week. Blood and gastric samples were collected at the end of the study. RESULTS: In vitro, at intact pH 4, mean inhibitory zone diameters of 8.5 mm and 13 mm were noted at concentrations of 5× and 10× of L. plantarum B7 supernatant disks, respectively. At adjusted pH 7, L. plantarum B7 supernatants at concentrations of 5× and 10× yielded mean inhibitory zone diameters of 6.5 mm and 11 mm, respectively. In the in vitro study, in group 2, stomach histopathology revealed mild to moderate H. pylori colonization and inflammation. The level of gastric MDA and epithelial cell apoptosis were significantly increased compared with group 1. The serum TNF-α level was significant decreased in group 3 compared with group 2 (P < 0.05). In addition, L. plantarum B7 treatments resulted in a significant improvement in stomach pathology, and decreased gastric MDA level and apoptotic epithelial cells

  15. Distinctive Intestinal Lactobacillus Communities in 6-Month-Old Infants From Rural Malawi and Southwestern Finland.

    PubMed

    Aakko, Juhani; Endo, Akihito; Mangani, Charles; Maleta, Kenneth; Ashorn, Per; Isolauri, Erika; Salminen, Seppo

    2015-12-01

    Our aim was to compare the composition and diversity of Lactobacillus microbiota in infants living in Malawi and Southwestern Finland. The composition and diversity of the Lactobacillus group was analyzed in the feces of healthy 6-month-old infants living in rural Malawi (n = 44) and Southwestern Finland (n = 31), using the quantitative polymerase chain reaction method and PCR-denaturing gradient gel electrophoresis fingerprinting. Malawian infants had higher counts of lactobacilli than their Finnish counterparts (7.45 log cells/g vs 6.86 log cells/g, P < 0.001, respectively) and the Lactobacillus community was richer and more diverse in the Malawian infants. Leuconostoc citreum and Weissella confusa were the predominant species in both study groups, but Malawian infants were more often colonized by these species (100% vs 74.2%, P < 0.001; 95.5% vs 41.9%, P < 0.001, respectively). Moreover, Lactobacillus ruminis, Lactobacillus gasseri, Lactobacillus acidophilus, and Lactobacillus mucosae were detected more often in the Malawian infants (59.1% vs 0.0%, P < 0.001; 38.6% vs 9.7%, P = 0.004; 29.5% vs 0.0%, P < 0.001; 22.7% vs 3.2%, P = 0.017, respectively). Lactobacillus casei group species, however, were only detected in the Finnish infants. Malawian infants have a more abundant Lactobacillus microbiota with a distinct composition compared with Finnish infants. The environment, including diet and hygiene, may be among the factors influencing these differences.

  16. Anti-H-Y responses of H-2b mutant mice.

    PubMed

    Simpson, E; Gordon, R D; Chandler, P R; Bailey, D

    1978-10-01

    Two strains of H-2b mutant mice, H-2ba and H-2bf, in which the mutational event took place at H-2K, make anti-H-Y cytotoxic T cell responses which are H-2-restricted, Db-associated and indistinguishable in target cell specificity from those of H-2b mice. Thus, alteration of the H-2K molecule affects neither the Ir gene controlling the response, nor the associative antigen. On the other hand, one H-2Db mutant strain, H-2bo, although it makes a good anti-H-Y cytotoxic response, shows target cell specificity restricted to its own Dbo antigen(s), and neither H-2b, H-2ba or H-2bf anti-H-Y cytotoxic cells kill H-2bo male target cells. Thus, the alteration of the H-2Db molecule does not affect the Ir gene of H-2b mice, but it does alter the H-2Db-associative antigen.

  17. Rapid molecular identification and characteristics of Lactobacillus strains.

    PubMed

    Markiewicz, L H; Biedrzycka, E; Wasilewska, E; Bielecka, M

    2010-09-01

    Eleven type strains and 24 Lactobacillus isolates, preliminarily classified to the species due to phenotypic features, were investigated. Standard methods of identification with species-specific PCRs and typing with PFGE (with ApaI, NotI and SmaI restriction enzymes) allowed us to distinguish 16 unique strains belonging to 5 species (L. acidophilus, L. delbrueckii ssp. bulgaricus, L. plantarum, L. rhamnosus, L. salivarius). Alternative approach with 16S-23S rDNA ARDRA identification (with merely two restrictases, BsuRI and TaqI) and PCR-based typing (RAPD with two random- and rep-PCR with (GTG)(5) primers) showed to be more discriminative, i.e. 21 unique strains were classified in the same species as above. As a result, 7 out of 24 phenotypically species-assigned isolates were reclassified. The alternative procedure of rapid identification and typing of Lactobacillus isolates appeared to be equally effective and shortened from 1 week to 2-3 d (in comparison to the standard methods).

  18. Protective effect of surface layer proteins isolated from four Lactobacillus strains on hydrogen-peroxide-induced HT-29 cells oxidative stress.

    PubMed

    Zhao, Bin-Bin; Meng, Jun; Zhang, Qiu-Xiang; Kang, Ting-Ting; Lu, Rong-Rong

    2017-09-01

    The objective of this study was to explore the antioxidant effect of the surface layer proteins (SLPs) and their mechanism. We investigated four SLPs which were extracted from L. casei zhang, L. rhamnosus, L. gasseri and L. acidophilus NCFM respectively using LiCl. The protective effect of SLPs on H 2 O 2 -induced HT-29 cells oxidative injury was investigated. As results, SLPs (100μg/mL) could significantly mitigate HT-29 cells cytotoxicity, improve the activities of total antioxidant capacity (T-AOC), catalase (CAT) and superoxide dismutase (SOD), decrease the contents of malondialdehyde (MDA) and lactate dehydrogenase (LDH), compared with H 2 O 2 -induced group (P<0.05). Furthermore, SLPs were also shown to attenuate the apoptosis rate (10.94-24.03%, P<0.01), suppress the elevation of intracellular reactive oxygen species (ROS) and calcium levels, restore mitochondrial membrane potential (MMP) and block the activation of apoptosis-related proteins of caspase-3 and caspase-9 (P<0.05). Considering all the parameters analyzed, we concluded that Lactobacillus SLPs play an essential role in the antioxidant capacity of HT-29 cells induced by H 2 O 2 , and the mechanism could be attributed to SLPs' ability to enhance the activity of the intracellular antioxidant enzyme system, reduce ROS accumulation and to inhibit apoptosis by regulating mitochondrial pathway. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Lactobacillus plantarum culture supernatants improve intestinal tissue exposed to deoxynivalenol.

    PubMed

    Maidana, L G; Gerez, J; Pinho, F; Garcia, S; Bracarense, A P F L

    2017-10-02

    In the present study, histological, morphometrical and ultrastructural analysis were performed to investigate intestinal mucosa changes in piglets exposed to deoxynivalenol alone or associated with two strains of Lactobacillus plantarum and the respective culture supernatants. Jejunal explants were incubated for 4h in culture medium with a) only culture medium (DMEM, control group), b) deoxynivalenol (DON, 10μM), c) heat-inactivated Lactobacillus plantarum strain1 - LP1 (1.1×10 8 CFU/ml) plus DON, d) heat-inactivated Lactobacillus plantarum strain2-LP2 (2.0×10 9 CFU/ml) plus DON, e) heat-inactivated Lactobacillus plantarum strain1 culture supernatant (CS1) plus DON, and f) heat-inactivated Lactobacillus plantarum strain1 culture supernatant (CS1) plus DON. Explants exposed to DON and DON plus LP1 and LP2 showed a significant increase in histological changes (mainly villi atrophy and apical necrosis) and a significant decrease in villi height when compared to unexposed explants. However, explants treated with CS1+DON and CS2+DON remained similar to the control group both in histological and morphometrical aspects. DON also induced a significant decrease in goblet cell density compared to control whereas CS1+DON treatment induced an increase in the number of goblet cells in comparison to DON explants. In addition, ultrastructural assessment showed control, CS1+DON and CS2+DON explants with well delineated finger shape villi, meanwhile DON-treated, LP1+DON and LP2+DON explants showed a severe villi atrophy with leukocytes exudation on the intestinal surface. Taken together, our results indicate that the culture supernatant treatment reduced the toxic effects induced by DON on intestinal tissue and may contribute as an alternative strategy to reduce mycotoxin toxicity. Copyright © 2017 Elsevier GmbH. All rights reserved.

  20. 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

  1. Purification and characterization of novel antifungal compounds from the sourdough Lactobacillus plantarum strain 21B.

    PubMed

    Lavermicocca, P; Valerio, F; Evidente, A; Lazzaroni, S; Corsetti, A; Gobbetti, M

    2000-09-01

    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.

  2. Absolute Enumeration of Probiotic Strains Lactobacillus acidophilus NCFM® and Bifidobacterium animalis subsp. lactis Bl-04 ® via Chip-Based Digital PCR.

    PubMed

    Hansen, Sarah J Z; Morovic, Wesley; DeMeules, Martha; Stahl, Buffy; Sindelar, Connie W

    2018-01-01

    The current standard for enumeration of probiotics to obtain colony forming units by plate counts has several drawbacks: long time to results, high variability and the inability to discern between bacterial strains. Accurate probiotic cell counts are important to confirm the delivery of a clinically documented dose for its associated health benefits. A method is described using chip-based digital PCR (cdPCR) to enumerate Bifidobacterium animalis subsp. lactis Bl-04 and Lactobacillus acidophilus NCFM both as single strains and in combination. Primers and probes were designed to differentiate the target strains against other strains of the same species using known single copy, genetic differences. The assay was optimized to include propidium monoazide pre-treatment to prevent amplification of DNA associated with dead probiotic cells as well as liberation of DNA from cells with intact membranes using bead beating. The resulting assay was able to successfully enumerate each strain whether alone or in multiplex. The cdPCR method had a 4 and 5% relative standard deviation (RSD) for Bl-04 and NCFM, respectively, making it more precise than plate counts with an industry accepted RSD of 15%. cdPCR has the potential to replace traditional plate counts because of its precision, strain specificity and the ability to obtain results in a matter of hours.

  3. Salivary Actinomyces naeslundii genospecies 2 and Lactobacillus casei levels predict pregnancy outcomes.

    PubMed

    Dasanayake, Ananda P; Li, Yihong; Wiener, Howard; Ruby, John D; Lee, Men-Jean

    2005-02-01

    Gravida's poor periodontal health is emerging as a modifiable independent risk factor for preterm delivery and low birth weight. To test the hypothesis that oral bacteria other than periodontal pathogens are also associated with pregnancy outcomes, specific oral bacterial levels measured during pregnancy were evaluated in relation to gestational age and birth weight while controlling for demographic, medical, and dental variables. The study population consisted of 297 predominantly African- American women who were pregnant for the first time. The salivary bacterial levels evaluated were Streptococcus mutans, Streptococcus sobrinus, Streptococcus sanguinus, Lactobacillus acidophilus, Lactobacillus casei, Actinomyces naeslundii genospecies (gsp) 1 and 2, total streptococci, and total cultivable organisms. For 1 unit increase in log(10) A. naeslundii gsp 2 levels, there was a 60 gm decrease in birth weight (beta = -59.7 g; SE = 29.1; P = 0.04), and a 0.17 week decrease in gestational age (beta = -0.17 wk; SE = 0.09; P = 0.05). In contrast, per 1 unit increase in log(10) L. casei levels, there was a 42 gm increase in birth weight (beta = 42.2 g; SE = 19.3; P = 0.03), and a 0.13 week increase in gestational age (beta = 0.13 week; SE = 0.06; P = 0.04). We conclude that other oral bacterial species can also be related to pregnancy outcomes in addition to previously reported periodontal pathogens. These organism levels may not only predict poor pregnancy outcomes, but also be used as modifiable risk factors in reducing prematurity and low birth weight.

  4. Characterization of Lactobacillus salivarius strains B37 and B60 capable of inhibiting IL-8 production in Helicobacter pylori-stimulated gastric epithelial cells.

    PubMed

    Panpetch, Wimonrat; Spinler, Jennifer K; Versalovic, James; Tumwasorn, Somying

    2016-10-18

    Interleukin (IL)-8 is the key agent for initiating an inflammatory response to infection with Helicobacter pylori. Some strains of Lactobacillus spp. are known to colonize the stomach and suppress inflammation caused by H. pylori. In this study, we characterized two gastric-derived lactobacilli, Lactobacillus salivarius (LS) strains B37 and B60, capable of inhibiting H. pylori-induced IL-8 production by gastric epithelial cells. Conditioned media from LS-B37 and LS-B60 suppressed H. pylori-induced IL-8 production and mRNA expression from AGS cells without inhibiting H. pylori growth. These conditioned media suppressed the activation of NF-κB but did not suppress c-Jun activation. IL-8 inhibitory substances in conditioned media of LS-B37 and LS-B60 are heat-stable and larger than 100 kDa in size. The inhibitory activity of LS-B37 was abolished when the conditioned medium was treated with α-amylase but still remained when treated with either proteinase K, trypsin, lipase or lysozyme. The activity of LS-B60 was abolished when the conditioned medium was treated with either amylase or proteinase K but still remained when treated with lysozyme. Treatment with lipase and trypsin also significantly affected the inhibitory activity of LS-B60 although the conditioned medium retained IL-8 suppression statistically different from media control. These results suggest that L. salivarius strains B37 and B60 produce different immunomodulatory factors capable of suppressing H. pylori-induced IL-8 production from gastric epithelial cells. Our results suggest that the large, heat-stable immunomodulatory substance(s) present in the LCM of LS-B37 is a polysaccharide, while the one(s) of LS-B60 is either complex consisting of components of polysaccharide, lipid and protein or includes multiple components such as glycoprotein and lipoprotein.

  5. Gamma-amino butyric acid (GABA) synthesis of Lactobacillus in fermentation of defatted rice bran extract

    NASA Astrophysics Data System (ADS)

    Dat, Lai Quoc; Ngan, Tran Thi Kim; Nu, Nguyen Thi Xuan

    2017-09-01

    This research focused on the synthesis of GABA by Lactobacillus bacteria in fermentation of defatted rice bran extract without adding glutamate. Two strains of Lactobacillus were investigated into capacity of GABA synthesis. Result indicates that, Lactobacillus brevis VTCC - B - 454 exhibited the higher capacity of GABA synthesis in fermentation of defatted rice bran extract than that of Lactobacillus plantarum VTCC - B - 890. Total dissolved solid (TDS), free amino acids (AA) and reducing sugar (RS) contents in fermentation of defatted rice bran extract with two strains also significantly decreased. At pH 5 and 9 %w/w of TDS content in defatted rice bran extract, Lactobacillus brevis VTCC - B - 454 accumulated 2,952 ppm of GABA in 24 hours of fermentation. The result implies that fermentation with Lactobacillus brevis VTCC - B - 454 can be applied for GABA production from defatted rice bran extract.

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

    USDA-ARS?s Scientific Manuscript database

    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 ...

  7. Effect of salt nutrients on mannitol production by Lactobacillus intermedius NRRL B-3693.

    PubMed

    Saha, Badal C

    2006-10-01

    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 liter in pH-controlled fermentation at 5.0 at 37 degrees C were evaluated using a fractional factorial design. Only manganese sulfate was found to be essential for mannitol production. Added manganese sulfate concentration of 0.033 g/l was found to support maximum production. The bacterium produced 200.6 +/- 0.2 g mannitol, 61.9 +/- 0.1 g lactic acid, and 40.4 +/- 0.3 g acetic acid from 300 g fructose per liter in 67 h.

  8. Molecular mechanisms underlying protection against H9N2 influenza virus challenge in mice by recombinant Lactobacillus plantarum with surface displayed HA2-LTB.

    PubMed

    Jiang, Yanlong; Yang, Guilian; Wang, Qi; Wang, Zhannan; Yang, Wentao; Gu, Wei; Shi, Chunwei; Wang, Jianzhong; Huang, Haibin; Wang, Chunfeng

    2017-10-10

    It has been considered that the Avian influenza virus (AIV) causes severe threats to poultry industry. In this study, we constructed a series of recombinant Lactobacillus plantarum (L. plantarum) with surface displayed hemagglutinin subunit 2 (HA2) alone or together with heat-labile toxin B subunit (LTB) from enterotoxigenic Escherichia coli. Balb/c mice were used as model to evaluate the protective effects of recombinant L. plantarum strains against H9N2 subtype challenge. The results showed that the presence of LTB significantly increased the percentages of CD3 + CD4 + IL-4 + , CD3 + CD4 + IFN-γ + and CD3 + CD4 + IL-17 + T cells, as well as CD3 + CD8 + IFN-γ + T cells in spleen and MLNs determined by Fluorescence-Activated Cell Sorting assay. Similar increased production of serum IFN-γ was also confirmed by enzyme linked immunosorbent assay (ELISA). The L. plantarum with surface displayed HA2-LTB also dramatically increased the percentages of B220 + IgA + B cells in peyer patch, in consistent with elevated production of mucosal SIgA antibody determined by ELISA. Finally, the orally administrated HA2-LTB expressing strain efficiently protected mice against H9N2 subtype AIV challenge shown by increased survival percentages, body weight gains and decreased lung lesions in histopathologic analysis. In conclusion, this study provides more detail mechanisms underlying the adjuvant effects of LTB on heterologous antigen produced in recombinant lactic acid bacteria. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Proteomic analyses of ethanol tolerance in Lactobacillus buchneri NRRL B-30929.

    PubMed

    Liu, Siqing

    2014-11-01

    The Lactobacillus buchneri NRRL B-30929 strain, isolated from a fuel ethanol (EtOH) production facility, exhibits high tolerance to environmental EtOH concentrations. This study aimed to identify proteins produced by B-30929 in response to environmental EtOH. Cellular proteins expressed by B-30929 growing in media with 10 versus 0% EtOH were compared by 2DE, followed by in-gel digestion and MALDI-MS analyses. Twenty EtOH responsive proteins were identified. These include a proline-specific peptidase (Lbuc_1852); a membrane protein (Lbuc_0921), two general stress-related proteins including a 10 kDa chaperonin (GroESL Lbuc_1359) and a 29 kDa member of the HK 97 family (Lbuc_1523); metabolic enzymes involving redox potential balances (Lbuc_2051 and Lbuc_0522) and carbohydrate fermentation (Lbuc_1319 and Lbuc_2157); nitrogen, amino acid, and fatty acid metabolism proteins (Lbuc_1994, Lbuc_0446, Lbuc_0858, Lbuc_0707, and Lbuc_0787). These changes suggested B-30929 cells respond to EtOH by degradation of available proteins and fatty acids and increased production of specific enzymes and molecular chaperons. These results can be used to guide genetic modifications to increase EtOH tolerance in industrial biocatalysts. The data have been deposited to World-2DPAGE (http://world-2dpage.expasy.org/repository/0068/; username liu, password 1h8d6Mg1). © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Reclassification of Lactobacillus kefirgranum Takizawa et al. 1994 as Lactobacillus kefiranofaciens subsp. kefirgranum subsp. nov. and emended description of L. kefiranofaciens Fujisawa et al. 1988.

    PubMed

    Vancanneyt, M; Mengaud, J; Cleenwerck, I; Vanhonacker, K; Hoste, B; Dawyndt, P; Degivry, M C; Ringuet, D; Janssens, D; Swings, J

    2004-03-01

    Fourteen homofermentative lactic acid bacteria that were isolated from kefir grains and kefir fermented milks were assigned to either Lactobacillus kefiranofaciens or Lactobacillus kefirgranum, based on their characteristic morphotypes, phenotypic features and SDS-PAGE profiles of whole-cell proteins. Further genotypic analyses on representative strains from both taxa demonstrated that L. kefiranofaciens and L. kefirgranum share 100 % 16S rDNA sequence similarity and belong phylogenetically to the Lactobacillus acidophilus species group. DNA-DNA binding values of >79 % and analogous DNA G+C contents of 37-38 mol% showed that the strains studied belonged to one species: L. kefirgranum is a later synonym of L. kefiranofaciens. An emended description is proposed for L. kefiranofaciens. Due to the specific morphological and biochemical characteristics of these taxa in kefir grain formation, it is proposed that L. kefirgranum should be reclassified as L. kefiranofaciens subsp. kefirgranum subsp. nov.

  11. Evaluation of tyrosinase inhibitory and antioxidant activities of Angelica dahurica root extracts for four different probiotic bacteria fermentations.

    PubMed

    Wang, Guey-Horng; Chen, Chih-Yu; Tsai, Teh-Hua; Chen, Ching-Kuo; Cheng, Chiu-Yu; Huang, Yi-Hsin; Hsieh, Min-Chi; Chung, Ying-Chien

    2017-06-01

    Angelica dahurica root (ADR), which shows strong antioxidant activity, is used in Chinese medicine. This study evaluated the tyrosinase inhibitory and antioxidant activities of ADR extracts fermented by four different probiotic bacteria: Bifidobacterium bifidum, Bifidobacterium lactis, Lactobacillus acidophilus, and Lactobacillus brevis. The ADR was first extracted using distilled water, 70% ethanol, and ethyl acetate, and then fermented by probiotic bacteria. The physiological characteristics of these fermented extracts, namely the antityrosinase activity, antioxidant activity, phenolic composition, and phenolic content, were evaluated and compared with those of unfermented extracts. Results showed that the water extracts after fermentation by probiotic bacteria exhibited the most favorable physiological characteristics. Among the extracts fermented by these probiotic bacteria, L. acidophilus-fermented ADR extract showed the most favorable physiological characteristics. The optimal IC 50 values for antityrosinase activity, DPPH radical scavenging activity, and reducing power for L. acidophilus-fermented ADR extract were 0.07 ± 0.03, 0.12 ± 0.01, and 0.68 ± 0.06 mg/mL, respectively. Furthermore, the physiological activities of fermented extracts were considerably higher than those of unfermented extracts. The tyrosinase inhibition and melanin content of B16F10 melanoma cells, and cytotoxicity effects of the fermented ADR extracts on B16F10 cells were also evaluated. We found that the L. acidophilus-fermented ADR extract at 1.5 mg/mL showed significant cellular antityrosinase activity with low melanin production in B16F10 cells and was noncytotoxic to B16F10 cells. Among all probiotic bacteria, water-extracted ADR fermented by L. acidophilus for 48 h was found to be the best skincare agent or antioxidant agent. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  12. Functional crosstalk between histone H2B ubiquitylation and H2A modifications and variants.

    PubMed

    Wojcik, Felix; Dann, Geoffrey P; Beh, Leslie Y; Debelouchina, Galia T; Hofmann, Raphael; Muir, Tom W

    2018-04-11

    Ubiquitylation of histone H2B at lysine residue 120 (H2BK120ub) is a prominent histone posttranslational modification (PTM) associated with the actively transcribed genome. Although H2BK120ub triggers several critical downstream histone modification pathways and changes in chromatin structure, less is known about the regulation of the ubiquitylation reaction itself, in particular with respect to the modification status of the chromatin substrate. Here we employ an unbiased library screening approach to profile the impact of pre-existing chromatin modifications on de novo ubiquitylation of H2BK120 by the cognate human E2:E3 ligase pair, UBE2A:RNF20/40. Deposition of H2BK120ub is found to be highly sensitive to PTMs on the N-terminal tail of histone H2A, a crosstalk that extends to the common histone variant H2A.Z. Based on a series of biochemical and cell-based studies, we propose that this crosstalk contributes to the spatial organization of H2BK120ub on gene bodies, and is thus important for transcriptional regulation.

  13. H2-K(b) and H2-D(b) regulate cerebellar long-term depression and limit motor learning.

    PubMed

    McConnell, Michael J; Huang, Yanhua H; Datwani, Akash; Shatz, Carla J

    2009-04-21

    There are more than 50 class I MHC (MHCI) molecules in the mouse genome, some of which are now known to be expressed in neurons; however, the role of classical MHCI molecules in synaptic plasticity is unknown. We report that the classical MHCI molecules, H2-K(b) and H2-D(b), are co-expressed by Purkinje cells (PCs). In the cerebellum of mice deficient for both H2-K(b) and H2-D(b) (K(b)D(b-/-)), there is a lower threshold for induction of long-term depression (LTD) at parallel fiber to PC synapses. This change may be a result of additional glutamate release observed at K(b)D(b-/-) CF to PC synapses, which are thought to "train" the cerebellar circuit. A behavioral correlate of cerebellar LTD is motor learning; acquisition and retention of a Rotarod behavioral task is significantly better in K(b)D(b-/-) mice than in WT cohorts. These physiological and behavioral phenotypes in K(b)D(b-/-) mice reveal a surprising role for classical MHCI molecules in synaptic plasticity and motor learning.

  14. H2MBH2 and M(μ-H)2BH2 Molecules Isolated in Solid Argon: Interelement M-B and M-H-B Bonds (M = Ge, Sn).

    PubMed

    Zhao, Jie; Beckers, Helmut; Huang, Tengfei; Wang, Xuefeng; Riedel, Sebastian

    2018-02-19

    Laser-ablated boron atoms react with GeH 4 molecules to form novel germylidene borane H 2 GeBH 2 , which undergoes a photochemical rearrangement to the germanium tetrahydroborate Ge(μ-H) 2 BH 2 upon irradiation with light of λ = 405 nm. For comparison, the boron atom reactions with SnH 4 only gave the tin tetrahydroborate Sn(μ-H) 2 BH 2 . Infrared matrix-isolation spectroscopy with deuterium substitution and the state-of-the-art quantum-chemical calculations are used to identify these species in solid argon. A planar structure of H 2 GeBH 2 with an electron-deficient B-Ge bond with a partial multiple bond character (bond order = 1.5) is predicted by quantum-chemical calculations. In the case of M(μ-H) 2 BH 2 (M = Ge, Sn) two 3c-2e B-H-M hydrogen bridged bonds are formed by donation of electrons from the B-H σ-bonds into empty p-orbitals of M.

  15. Evaluation of agrowastes as immobilizers for probiotics in soy milk.

    PubMed

    Teh, Sue-Siang; Ahmad, Rosma; Wan-Abdullah, Wan-Nadiah; Liong, Min-Tze

    2009-11-11

    The objective of this study was to evaluate agricultural wastes as immobilizers for probiotics in liquid foods, such as soy milk. Probiotic strains were initially evaluated for acid and bile tolerance and the ability to produce alpha-galactosidase. Rinds of durian, mangosteen, and jackfruit were dried, ground, and sterilized prior to immobilization of selected strains ( Lactobacillus acidophilus FTDC 1331, L. acidophilus FTDC 2631, L. acidophilus FTDC 2333, L. acidophilus FTDC 1733, and Lactobacillus bulgaricus FTCC 0411). Immobilized cells were inoculated into soy milk, and growth properties were evaluated over 168 h at 37 degrees C. Soy milk containing free cells without agrowastes was used as the control. Immobilized probiotics showed increased growth, greater reduction of stachyose, sucrose, and glucose, higher production of lactic and acetic acids, and lower pH in soy milk compared to the control. The results illustrated that agrowastes could be used for the immobilization of probiotics with enhanced growth, utilization of substrates, and production of organic acids.

  16. Characterization of vaginal Lactobacillus species by rplK -based multiplex qPCR in Russian women.

    PubMed

    Demkin, Vladimir V; Koshechkin, Stanislav I

    2017-10-01

    We describe a multiplex qPCR assay for identification and quantitative assessment of a set of vaginal Lactobacillus species, including L. acidophilus, L. crispatus, L. gasseri, L. helveticus, L. iners, and L. jensenii. The assay extends the previously developed qPCR method for Lactobacillus detection and total quantification based on targeting the rplK gene. Both assays use only single pair of primers and a set of probes combined in three reactions, comprising a vaginal Lactobacillus diagnostic assay panel. The utility of the diagnostic panel was evaluated by analyzing of vaginal swab specimens from 145 patients with different status of vaginal health. Most frequently, only one Lactobacillus species was dominant (68,9%), mostly L. crispatus (18,6%) or L. iners (33,1%), but two or three Lactobacillus species were also being simultaneously detected (24,9%). The diagnostic panel will facilitate investigations of the role of Lactobacillus species in the health of the female reproductive system and promote studies of variability of the vaginal microbiota. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Exploring the ameliorative potential of probiotic Dahi containing Lactobacillus acidophilus and Bifidobacterium bifidum on dextran sodium sulphate induced colitis in mice.

    PubMed

    Jadhav, Sagar R; Shandilya, Umesh Kr; Kansal, Vinod K

    2013-02-01

    Conventional medical therapies for ulcerative colitis (UC) are still limited due to the adverse side effects like dose-dependent diarrhoea and insufficient potency to keep in remission for long-term periods. So, new alternatives that provide more effective and safe therapies for ulcerative colitis are constantly being sought. In the present study, probiotic LaBb Dahi was selected for investigation of its therapeutic effect on DSS-induced colitis model in mice. LaBb Dahi was prepared by co-culturing Dahi culture of Lactococci along with selected strain of Lactobacillus acidophilus LaVK2 and Bifidobacterium bifidum BbVK3 in buffalo milk. Four groups of mice (12 each) were fed for 17 d with buffalo milk (normal control), buffalo milk plus DSS (Colitis control), Dahi plus DSS, and LaBb Dahi plus DSS, respectively, with basal diet. The disease activity scores, weight loss, organ weight, colon length, myeloperoxidase (MPO) and β-glucoronidase activity was assessed, and the histopathological picture of the colon of mice was studied. All colitis control mice evidenced significant increase in MPO, β-glucoronidase activity and showed high disease activity scores along with histological damage to colonic tissue. Feeding with LaBb Dahi offered significant reduction in MPO activity, β-glucoronidase activity and improved disease activity scores. We found significant decline in length of colon, organ weight and body weight in colitis induced controls which were improved significantly by feeding LaBb Dahi. The present study suggests that LaBb Dahi can be used as a potential nutraceutical intervention to combat UC related changes and may offer effective adjunctive treatment for management of UC.

  18. Both IIC and IID Components of Mannose Phosphotransferase System Are Involved in the Specific Recognition between Immunity Protein PedB and Bacteriocin-Receptor Complex.

    PubMed

    Zhou, Wanli; Wang, Guohong; Wang, Chunmei; Ren, Fazheng; Hao, Yanling

    2016-01-01

    Upon exposure to exogenous pediocin-like bacteriocins, immunity proteins specifically bind to the target receptor of the mannose phosphotransferase system components (man-PTS IIC and IID), therefore preventing bacterial cell death. However, the specific recognition of immunity proteins and its associated target receptors remains poorly understood. In this study, we constructed hybrid receptors to identify the domains of IIC and/or IID recognized by the immunity protein PedB, which confers immunity to pediocin PA-1. Using Lactobacillus plantarum man-PTS EII mutant W903, the IICD components of four pediocin PA-1-sensitive strains (L. plantarum WQ0815, Leuconostoc mesenteroides 05-43, Lactobacillus salivarius REN and Lactobacillus acidophilus 05-172) were respectively co-expressed with the immunity protein PedB. Well-diffusions assays showed that only the complex formed by LpIICD from L. plantarum WQ0815 with pediocin PA-1 could be recognized by PedB. In addition, a two-step PCR approach was used to construct hybrid receptors by combining LpIIC or LpIID recognized by PedB with the other three heterologous IID or IIC compounds unrecognized by PedB, respectively. The results showed that all six hybrid receptors were recognized by pediocin PA-1. However, when IIC or IID of L. plantarum WQ0815 was replaced with any corresponding IIC or IID component from L. mesenteroides 05-43, L. salivarius REN and L. acidophilus 05-172, all the hybrid receptors could not be recognized by PedB. Taken altogether, we concluded that both IIC and IID components of the mannose phosphotransferase system play an important role in the specific recognition between the bacteriocin-receptor complex and the immunity protein PedB.

  19. High-fibre diet and Lactobacillus paracasei B21060 in symptomatic uncomplicated diverticular disease.

    PubMed

    Lahner, Edith; Esposito, Gianluca; Zullo, Angelo; Hassan, Cesare; Cannaviello, Claudio; Paolo, Maria Carla Di; Pallotta, Lorella; Garbagna, Nicoletta; Grossi, Enzo; Annibale, Bruno

    2012-11-07

    To investigate in symptomatic uncomplicated diverticular disease the efficacy of symbiotics associated with a high-fibre diet on abdominal symptoms. This study was a multicentre, 6-mo randomized, controlled, parallel-group intervention with a preceding 4-wk washout period. Consecutive outpatients with symptomatic uncomplicated diverticular disease, aged 40-80 years, evaluated in 4 Gastroenterology Units, were enrolled. Symptomatic uncomplicated diverticular disease patients were randomized to two treatment arms A or B. Treatment A (n = 24 patients) received 1 symbiotic sachet Flortec(©) (Lactobacillus paracasei B21060) once daily plus high-fibre diet for 6 mo. Treatment B (n = 21 patients) received high-fibre diet alone for 6 mo. The primary endpoint was regression of abdominal symptoms and change of symptom severity after 3 and 6 mo of treatment. In group A, the proportion of patients with abdominal pain < 24 h decreased from 100% at baseline to 35% and 25% after 3 and 6 mo, respectively (P < 0.001). In group B the proportion of patients with this symptom decreased from 90.5% at baseline to 61.9% and 38.1% after 3 and 6 mo, respectively (P = 0.001). Symptom improvement became statistically significant at 3 and 6 mo in group A and B, respectively.The proportion of patients with abdominal pain >24 h decreased from 60% to 20% then 5% after 3 and 6 mo, respectively in group A (P < 0.001) and from 33.3% to 9.5% at both 3 and 6 mo in group B (P = 0.03). In group A the proportion of patients with abdominal bloating significantly decreased from 95% to 60% after 3 mo, and remained stable (65%) at 6-mo follow-up (P = 0.005) while in group B, no significant changes in abdominal bloating was observed (P = 0.11). After 6 mo of treatment, the mean visual analogic scale (VAS) values of both short-lasting abdominal pain (VAS, mean ± SD, group A: 4.6 ± 2.1 vs 2.2 ± 0.8, P = 0.02; group B: 4.6 ± 2.9 vs 2.0 ± 1.9, P = 0.03) and abdominal bloating (VAS, mean ± SD, group A: 5

  20. Nucleoplasmin Binds Histone H2A-H2B Dimers through Its Distal Face*

    PubMed Central

    Ramos, Isbaal; Martín-Benito, Jaime; Finn, Ron; Bretaña, Laura; Aloria, Kerman; Arizmendi, Jesús M.; Ausió, Juan; Muga, Arturo; Valpuesta, José M.; Prado, Adelina

    2010-01-01

    Nucleoplasmin (NP) is a pentameric chaperone that regulates the condensation state of chromatin extracting specific basic proteins from sperm chromatin and depositing H2A-H2B histone dimers. It has been proposed that histones could bind to either the lateral or distal face of the pentameric structure. Here, we combine different biochemical and biophysical techniques to show that natural, hyperphosphorylated NP can bind five H2A-H2B dimers and that the amount of bound ligand depends on the overall charge (phosphorylation level) of the chaperone. Three-dimensional reconstruction of NP/H2A-H2B complex carried out by electron microscopy reveals that histones interact with the chaperone distal face. Limited proteolysis and mass spectrometry indicate that the interaction results in protection of the histone fold and most of the H2A and H2B C-terminal tails. This structural information can help to understand the function of NP as a histone chaperone. PMID:20696766

  1. Evidence for the η(b)(2S) and observation of h(b)(1P)→η(b)(1S)γ and h(b)(2P)→η(b)(1S)γ.

    PubMed

    Mizuk, R; Asner, D M; Bondar, A; Pedlar, T K; Adachi, I; Aihara, H; Arinstein, K; Aulchenko, V; Aushev, T; Aziz, T; Bakich, A M; Bay, A; Belous, K; Bhardwaj, V; Bhuyan, B; Bischofberger, M; Bonvicini, G; Bozek, A; Bračko, M; Brodzicka, J; Browder, T E; Chekelian, V; Chen, A; Chen, P; Cheon, B G; Chilikin, K; Chistov, R; Cho, I-S; Cho, K; Choi, S-K; Choi, Y; Dalseno, J; Danilov, M; Doležal, Z; Drásal, Z; Drutskoy, A; Eidelman, S; Epifanov, D; Fast, J E; Gaur, V; Gabyshev, N; Garmash, A; Golob, B; Haba, J; Hara, T; Hayasaka, K; Hayashii, H; Horii, Y; Hoshi, Y; Hou, W-S; Hsiung, Y B; Hyun, H J; Iijima, T; Ishikawa, A; Itoh, R; Iwabuchi, M; Iwasaki, Y; Iwashita, T; Jaegle, I; Julius, T; Kang, J H; Kapusta, P; Kawasaki, T; Kim, H J; Kim, H O; Kim, J H; Kim, K T; Kim, M J; Kim, Y J; Kinoshita, K; Ko, B R; Koblitz, S; Kodyš, P; Korpar, S; Kouzes, R T; Križan, P; Krokovny, P; Kuhr, T; Kumita, T; Kuzmin, A; Kwon, Y-J; Lange, J S; Lee, S-H; Li, J; Libby, J; Liu, C; Liu, Y; Liu, Z Q; Liventsev, D; Louvot, R; Matvienko, D; McOnie, S; Miyabayashi, K; Miyata, H; Mohanty, G B; Mohapatra, D; Moll, A; Muramatsu, N; Mussa, R; Nakao, M; Natkaniec, Z; Ng, C; Nishida, S; Nishimura, K; Nitoh, O; Nozaki, T; Ohshima, T; Okuno, S; Olsen, S L; Onuki, Y; Pakhlov, P; Pakhlova, G; Park, C W; Park, H; Pestotnik, R; Petrič, M; Piilonen, L E; Poluektov, A; Röhrken, M; Sakai, Y; Sandilya, S; Santel, D; Sanuki, T; Sato, Y; Schneider, O; Schwanda, C; Senyo, K; Seon, O; Sevior, M E; Shapkin, M; Shen, C P; Shibata, T-A; Shiu, J-G; Shwartz, B; Sibidanov, A; Simon, F; Smerkol, P; Sohn, Y-S; Sokolov, A; Solovieva, E; Stanič, S; Starič, M; Sumihama, M; Sumiyoshi, T; Tanida, K; Tatishvili, G; Teramoto, Y; Tikhomirov, I; Trabelsi, K; Tsuboyama, T; Uchida, M; Uehara, S; Uglov, T; Unno, Y; Uno, S; Vanhoefer, P; Varner, G; Varvell, K E; Vinokurova, A; Vorobyev, V; Wang, C H; Wang, M-Z; Wang, P; Wang, X L; Watanabe, M; Watanabe, Y; Williams, K M; Won, E; Yabsley, B D; Yamaoka, J; Yamashita, Y; Yuan, C Z; Zhang, Z P; Zhilich, V

    2012-12-07

    We report the first evidence for the η(b)(2S) using the h(b)(2P)→η(b)(2S)γ transition and the first observation of the h(b)(1P)→η(b)(1S)γ and h(b)(2P)→η(b)(1S)γ transitions. The mass and width of the η(b)(1S) and η(b)(2S) are measured to be m(η(b)(1S))=(9402.4±1.5±1.8) MeV/c(2), m(η(b)(2S))=(9999.0±3.5(-1.9)(+2.8)) MeV/c(2), and Γ(η(b)(1S))=(10.8(-3.7-2.0)(+4.0+4.5)) MeV. We also update the h(b)(1P) and h(b)(2P) mass measurements. We use a 133.4 fb(-1) data sample collected at energies near the Υ(5S) resonance with the Belle detector at the KEKB asymmetric-energy e(+)e(-) collider.

  2. The H2A.Z/H2B dimer is unstable compared to the dimer containing the major H2A isoform.

    PubMed

    Placek, Brandon J; Harrison, L Nicole; Villers, Brooke M; Gloss, Lisa M

    2005-02-01

    The nucleosome, the basic fundamental repeating unit of chromatin, contains two H2A/H2B dimers and an H3/H4 tetramer. Modulation of the structure and dynamics of the nucleosome is an important regulation mechanism of DNA-based chemistries in the eukaryotic cell, such as transcription and replication. One means of altering the properties of the nucleosome is by incorporation of histone variants. To provide insights into how histone variants may impact the thermodynamics of the nucleosome, the stability of the heterodimer between the H2A.Z variant and H2B was determined by urea-induced denaturation, monitored by far-UV circular dichroism, intrinsic Tyr fluorescence intensity, and anisotropy. In the absence of stabilizing agents, the H2A.Z/H2B dimer is only partially folded. The stabilizing cosolute, trimethylamine-N-oxide (TMAO) was used to promote folding of the unstable heterodimer. The equilibrium stability of the H2A.Z/H2B dimer is compared to that of the H2A/H2B dimer. The equilibrium folding of both histone dimers is highly reversible and best described by a two-state model, with no detectable equilibrium intermediates populated. The free energies of unfolding, in the absence of denaturant, of H2A.Z/H2B and H2A/H2B are 7.3 kcal mol(-1) and 15.5 kcal mol(-1), respectively, in 1 M TMAO. The H2A.Z/H2B dimer is the least stable histone fold characterized to date, while H2A/H2B appears to be the most stable. It is speculated that this difference in stability may contribute to the different biophysical properties of nucleosomes containing the major H2A and the H2A.Z variant.

  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 μg/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 β-galactosidase and N-acetyl-β-glucosaminidase activities. It also did not produce carcinogenic enzymes such as β-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. PMID:26761499

  4. 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

  5. Production of B atoms and BH radicals from B2H6/He/H2 mixtures activated on heated W wires.

    PubMed

    Umemoto, Hironobu; Kanemitsu, Taijiro; Tanaka, Akihito

    2014-07-17

    B atoms and BH radicals could be identified by laser-induced fluorescence when B2H6/He/H2 mixtures were activated on heated tungsten wires. The densities of these radical species increased not only with the wire temperature but also with the partial pressure of H2. The densities in the presence of 0.026 Pa of B2H6 and 2.6 Pa of H2 were on the order of 10(11) cm(-3) both for B and BH when the wire temperature was 2000 K. Densities in the absence of a H2 flow were much smaller, suggesting that the direct production of these species on wire surfaces is minor. B and BH must be produced in the H atom shifting reactions, BH(x) + H → BH(x-1) + H2 (x = 1-3), in the gas phase, while H atoms are produced from H2 on wire surfaces. The B atom density increased monotonously with the H atom density, while the BH density showed saturation. These tendencies could be reproduced by simple modeling based on ab initio potential energy calculations and the transition-state theoretical calculations of the rate constants. The absolute densities could also be reproduced within a factor of 2.5.

  6. Influence of fructooligosaccharides on the fermentation profile and viable counts in a symbiotic low fat milk

    PubMed Central

    Oliveira, Ricardo P.S.; Casazza, Alessandro A.; Aliakbarian, Bahar; Perego, Patrizia; Converti, Attilio; Oliveira, Maricê N.

    2013-01-01

    This study evaluated the effects of prebiotics on fermentation profile and growth of Lactobacillus acidophilus, Lactobacillus bulgaricus, Lactobacillus rhamnosus, and Bifidobacterium lactis in co-cultures with Streptococcus thermophilus. Acidification rate and viability were positively influenced by the co-culture with B. lactis and by both inulin or oligofructose in low fat milk. PMID:24294233

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

    PubMed

    Bhandari, Praveen; Prabha, Vijay

    2015-07-01

    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. 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 10 [6] cfu/20 µl E. coli, and probiotic group was administered with Lactobacillus (10 [8] cfu/20 µl) for 10 consecutive days. In prophylactic group, the vagina was colonized with 10 consecutive doses of Lactobacillus (10 [8] cfu/20 µl). After 24 h, it was followed by 10 day intravaginal infection with E. coli (10 [6] cfu/20 µl) whereas for the therapeutic group vagina was colonized with (10 [6] cfu/20 µl) E. coli for 10 consecutive days, followed by 10 day intravaginal administration with Lactobacillus after 24 h. 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. Results indicated that Lactobacillus intermitted colonization of pathogenic strains that resulted in reinforcement of natural microflora and resurge fertility.

  8. Antibacterial activity of Lactobacillus spp. isolated from the feces of healthy infants against enteropathogenic bacteria.

    PubMed

    Davoodabadi, Abolfazl; Soltan Dallal, Mohammad Mehdi; Rahimi Foroushani, Abbas; Douraghi, Masoumeh; Sharifi Yazdi, Mohammad Kazem; Amin Harati, Farzaneh

    2015-08-01

    Lactobacilli are normal microflora of the gastrointestinal (GI) tract and are a heterogeneous group of lactic acid bacteria (LAB). Lactobacillus strains with Probiotic activity may have health Benefits for human. This study investigates the probiotic potential of Lactobacillus strains obtained from the feces of healthy infants and also explores antibacterial activity of Lactobacillus strains with probiotic potential against enteropathogenic bacteria. Fecal samples were collected from 95 healthy infants younger than 18 months. Two hundred and ninety Lactobacillus strains were isolated and assessed for probiotic potential properties including ability to survive in gastrointestinal conditions (pH 2.0, 0.3% oxgall), adherence to HT-29 cells and antibiotic resistance. Six strains including Lactobacillus fermentum (4 strains), Lactobacillus paracasei and Lactobacillus plantarum showed good probiotic potential and inhibited the growth of enteropathogenic bacteria including ETEC H10407, Shigella flexneri ATCC 12022, Shigella sonnei ATCC 9290, Salmonella enteritidis H7 and Yersinia enterocolitica ATCC 23715. These Lactobacillus strains with probiotic potential may be useful for prevention or treatment of diarrhea, but further in vitro and in vivo studies on these strains are still required. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Construction of recombinant Kluyveromyces marxianus UFV-3 to express dengue virus type 1 nonstructural protein 1 (NS1).

    PubMed

    Bragança, Caio Roberto Soares; Colombo, Lívia Tavares; Roberti, Alvaro Soares; Alvim, Mariana Caroline Tocantins; Cardoso, Silvia Almeida; Reis, Kledna Constancio Portes; de Paula, Sérgio Oliveira; da Silveira, Wendel Batista; Passos, Flavia Maria Lopes

    2015-02-01

    The yeast Kluyveromyces marxianus is a convenient host for industrial synthesis of biomolecules. However, despite its potential, there are few studies reporting the expression of heterologous proteins using this yeast. Here, we report expression of a dengue virus protein in K. marxianus for the first time. The dengue virus type 1 nonstructural protein 1 (NS1) was integrated into the K. marxianus UFV-3 genome at the LAC4 locus using an adapted integrative vector designed for high-level expression of recombinant protein in Kluyveromyces lactis. The NS1 gene sequence was codon-optimized to increase the level of protein expression in yeast. The synthetic gene was cloned in frame with K. lactis α-mating factor signal peptide, and the recombinant plasmid obtained was used to transform K. marxianus UFV-3 by electroporation. The transformed cells, selected in yeast extract peptone dextrose containing 200 μg mL(-1) Geneticin, were mitotically stable. Analysis of recombinant strains by RT-PCR and protein detection using blot analysis confirmed both transcription and expression of extracellular NS1 polypeptide. After induction with galactose, the NS1 protein was analyzed by sodium dodecyl sulfate-PAGE and immunogenic detection. Protein production was investigated under two conditions: with galactose and biotin pulses at 24-h intervals during 96 h of induction and without galactose and biotin supplementation. Protease activity was not detected in post-growth medium. Our results indicate that recombinant K. marxianus is a good host for the production of dengue virus NS1 protein, which has potential for diagnostic applications.

  10. Proteomic analyses of ethanol tolerance in Lactobacillus buchneri NRRL B-30929

    USDA-ARS?s Scientific Manuscript database

    The Lactobacillus buchneri NRRL B-30929 strain, isolated from a fuel ethanol production facility, exhibits high tolerance to environmental ethanol concentrations. This study aimed to identify proteins produced by B-30929 in response to environmental ethanol. Cellular proteins expressed by B-30929 gr...

  11. Resveratrol induces cellular senescence with attenuated mono-ubiquitination of histone H2B in glioma cells

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Gao, Zhen; Xu, Michael S.; Barnett, Tamara L.

    2011-04-08

    Research highlights: {yields} Resveratrol induces cellular senescence in glioma cell. {yields} Resveratrol inhibits mono-ubiquitination of histone H2B at K120. {yields} Depletion of RNF20, phenocopies the inhibitory effects of resveratrol. {yields} Mono-ubiquitination of histone H2B at K120 is a novel target of resveratrol. {yields} RNF20 inhibits cellular senescence in proliferating glioma cells. -- Abstract: Resveratrol (3,4',5-trihydroxy-trans-stilbene), a polyphenol naturally occurring in grapes and other plants, has cancer chemo-preventive effects and therapeutic potential. Although resveratrol modulates multiple pathways in tumor cells, how resveratrol or its affected pathways converge on chromatin to mediate its effects is not known. Using glioma cells as amore » model, we showed here that resveratrol inhibited cell proliferation and induced cellular hypertrophy by transforming spindle-shaped cells to enlarged, irregular and flatten-shaped ones. We further showed that resveratrol-induced hypertrophic cells expressed senescence-associated-{beta}-galactosidase, suggesting that resveratrol-induced cellular senescence in glioma cells. Consistent with these observations, we demonstrated that resveratrol inhibited clonogenic efficiencies in vitro and tumor growth in a xenograft model. Furthermore, we found that acute treatment of resveratrol inhibited mono-ubiquitination of histone H2B at K120 (uH2B) in breast, prostate, pancreatic, lung, brain tumor cells as well as primary human cells. Chronic treatment with low doses of resveratrol also inhibited uH2B in the resveratrol-induced senescent glioma cells. Moreover, we showed that depletion of RNF20, a ubiquitin ligase of histone H2B, inhibited uH2B and induced cellular senescence in glioma cells in vitro, thereby recapitulated the effects of resveratrol. Taken together, our results suggest that uH2B is a novel direct or indirect chromatin target of resveratrol and RNF20 plays an important role in inhibiting

  12. Lactobacilli Inactivate Chlamydia trachomatis through Lactic Acid but Not H2O2

    PubMed Central

    Gong, Zheng; Luna, Yesmin; Yu, Ping; Fan, Huizhou

    2014-01-01

    Lactobacillus species dominate the microbiome in the lower genital tract of most reproductive-age women. Producing lactic acid and H2O2, lactobacilli are believed to play an important role in prevention of colonization by and growth of pathogens. However, to date, there have been no reported studies characterizing how lactobacilli interact with Chlamydia trachomatis, a leading sexually transmitted bacterium. In this report, we demonstrate inactivation of C. trachomatis infectivity by culture media conditioned by Lactobacillus crispatus, L. gasseri and L. jensenii, known to be dominating organisms in the human vaginal microbiome. Lactobacillus still cultures produced lactic acid, leading to time- and concentration-dependent killing of C. trachomatis. Neutralization of the acidic media completely reversed chlamydia killing. Addition of lactic acid into Lactobacillus-unconditioned growth medium recapitulated the chlamydiacidal activity of conditioned media. The H2O2 concentrations in the still cultures were found to be comparable to those reported for the cervicovaginal fluid, but insufficient to inactivate chlamydiae. Aeration of Lactobacillus cultures by shaking markedly induced H2O2 production, but strongly inhibited Lactobacillus growth and lactic acid production, and thus severely affected acidification, leading to significantly reduced chlamydiacidal efficiency. These observations indicate lactobacilli inactivate chlamydiae primarily through maintaining acidity in a relatively hypoxic environment in the vaginal lumen with limited H2O2, which is consistent with the notion that women with higher vaginal pH are more prone to sexually transmitted C. trachomatis infection. In addition to lactic acid, formic acid and acetic acid also exhibited potent chlamydiacidal activities. Taken together, our findings imply that lowering the vaginal pH through engineering of the vaginal microbiome and other means will make women less susceptible to C. trachomatis infection. PMID

  13. The preliminary study of prebiotic potential of Polish wild mushroom polysaccharides: the stimulation effect on Lactobacillus strains growth.

    PubMed

    Nowak, Renata; Nowacka-Jechalke, Natalia; Juda, Marek; Malm, Anna

    2018-06-01

    According to the vast body of evidence demonstrating that the intestinal microbiota is undoubtedly linked with overall health, including cancer risk, searching for functional foods and novel prebiotic influencing on beneficial bacteria is necessary. The present study aimed to investigate the potential of polysaccharides from 53 wild-growing mushrooms to stimulate the growth of Lactobacillus acidophilus and Lactobacillus rhamnosus and to determine the digestibility of polysaccharide fractions. Mushroom polysaccharides were precipitated with ethanol from aqueous extracts. Determination of growth promoting activity of polysaccharides was performed in U-shaped 96-plates in an ELISA reader in relation to the reference strain of L. acidophilus and two clinical strains of L. rhamnosus. The digestibility of mushroom polysaccharides was investigated in vitro by exposing them to artificial human gastric juice. Obtained results revealed that fungal polysaccharides stimulate the growth of Lactobacillus strains stronger than commercially available prebiotics like inulin or fructooligosaccharides. Moreover, selected polysaccharides were subjected to artificial human gastric juice and remain undigested in more than 90%. Obtained results indicate that mushroom polysaccharides are able to pass through the stomach unchanged, reaching the colon and stimulating the growth of beneficial bacteria. Majority of 53 polysaccharide fractions were analysed for the first time in our study. Overall, our findings suggest that polysaccharide fractions from edible mushrooms might be useful in producing functional foods and nutraceuticals.

  14. Fe{sub 2}O{sub 3}-Bi{sub 2}O{sub 3}-B{sub 2}O{sub 3} glasses as lithium-free nonsilicate pH responsive glasses – Compatibility between pH responsivity and hydrophobicity

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hashimoto, Tadanori, E-mail: hasimoto@chem.mie-u.ac.jp; Hamajima, Mitsuaki; Ohta, Honami

    Highlights: • Fe{sub 2}O{sub 3}-rich FeBiB glasses show high pH sensitivity and short pH response time. • Bi{sub 2}O{sub 3}-rich FeBiB glasses show relatively high contact angle for water. • FeBiB glasses are lithium-free nonsilicate pH responsive ones. • pH responsivity and hydrophobicity are obtained for optimum glass compositions. - Abstract: Lithium silicate-based glasses have widely been used as commercially available pH glass electrodes. It was revealed that Ti{sup 3+}-containing titanophosphate (TiO{sub 2}-P{sub 2}O{sub 5}, TP) glasses are pH responsive as lithium-free nonsilicate glasses for the first time. TP glasses with the compatibility between pH responsivity and self-cleaning property weremore » obtained by the sequential post-annealing (oxidation and reduction) of as-prepared glasses. Bi{sub 2}O{sub 3}-B{sub 2}O{sub 3} (BiB) glasses are relatively hydrophobic and are expected to show anti-fouling effect. They are unsuitable for pH responsive glasses, because they have high electrical resistivity. In the present study, xFe{sub 2}O{sub 3}·yBi{sub 2}O{sub 3}·(100 − x − y)B{sub 2}O{sub 3} glasses (xFeyBiB, x = 0–20 mol%, y = 20–80 mol%) glasses were selected as new pH responsive glasses with hydrophobicity, because Fe{sub 2}O{sub 3} is a representative component for causing hopping conduction to the glasses. BiB glass did not show pH responsivity, whereas xFeyBiB glasses showed good pH responsivity. xFeyBiB glasses are lithium-free nonsilicate pH responsive ones as well as TP glasses. The electrical resistivity and pH response time decreased with increasing Fe{sub 2}O{sub 3} content. The pH repeatability for standard solutions increased with increasing Bi{sub 2}O{sub 3} content. Silicate glass (20Fe70BiSi) showed better pH responsivity but lower contact angle than those of borate glass (20Fe70BiB). pH sensitivity increased in order of TP glasses (about 80%), xFeyBiB glasses (about 90%) and commercial pH responsive glass (about 100

  15. Rapid identification of probiotic Lactobacillus species by multiplex PCR using species-specific primers based on the region extending from 16S rRNA through 23S rRNA.

    PubMed

    Kwon, Hyuk-Sang; Yang, Eun-Hee; Yeon, Seung-Woo; Kang, Byoung-Hwa; Kim, Tae-Yong

    2004-10-15

    This study aimed to develop a novel multiplex polymerase chain reaction (PCR) primer set for the identification of seven probiotic Lactobacillus species such as Lactobacillus acidophilus, Lactobacillus delbrueckii, Lactobacillus casei, Lactobacillus gasseri, Lactobacillus plantarum, Lactobacillus reuteri and Lactobacillus rhamnosus. The primer set, comprising of seven specific and two conserved primers, was derived from the integrated sequences of 16S and 23S rRNA genes and their rRNA intergenic spacer region of each species. It was able to identify the seven target species with 93.6% accuracy, which exceeds that of the general biochemical methods. The phylogenetic analyses, using 16S rDNA sequences of the probiotic isolates, also provided further support that the results from the multiplex PCR assay were trustworthy. Taken together, we suggest that the multiplex primer set is an efficient tool for simple, rapid and reliable identification of seven Lactobacillus species.

  16. Use of colony-based bacterial strain typing for tracking the fate of Lactobacillus strains during human consumption

    PubMed Central

    2009-01-01

    Background The Lactic Acid Bacteria (LAB) are important components of the healthy gut flora and have been used extensively as probiotics. Understanding the cultivable diversity of LAB before and after probiotic administration, and being able to track the fate of administered probiotic isolates during feeding are important parameters to consider in the design of clinical trials to assess probiotic efficacy. Several methods may be used to identify bacteria at the strain level, however, PCR-based methods such as Random Amplified Polymorphic DNA (RAPD) are particularly suited to rapid analysis. We examined the cultivable diversity of LAB in the human gut before and after feeding with two Lactobacillus strains, and also tracked the fate of these two administered strains using a RAPD technique. Results A RAPD typing scheme was developed to genetically type LAB isolates from a wide range of species, and optimised for direct application to bacterial colony growth. A high-throughput strategy for fingerprinting the cultivable diversity of human faeces was developed and used to determine: (i) the initial cultivable LAB strain diversity in the human gut, and (ii) the fate of two Lactobacillus strains (Lactobacillus salivarius NCIMB 30211 and Lactobacillus acidophilus NCIMB 30156) contained within a capsule that was administered in a small-scale human feeding study. The L. salivarius strain was not cultivated from the faeces of any of the 12 volunteers prior to capsule administration, but appeared post-feeding in four. Strains matching the L. acidophilus NCIMB 30156 feeding strain were found in the faeces of three volunteers prior to consumption; after taking the Lactobacillus capsule, 10 of the 12 volunteers were culture positive for this strain. The appearance of both Lactobacillus strains during capsule consumption was statistically significant (p < 0.05). Conclusion We have shown that genetic strain typing of the cultivable human gut microbiota can be evaluated using a high

  17. H2O2 production rate in Lactobacillus johnsonii is modulated via the interplay of a heterodimeric flavin oxidoreductase with a soluble 28 Kd PAS domain containing protein

    PubMed Central

    Valladares, Ricardo B.; Graves, Christina; Wright, Kaitlyn; Gardner, Christopher L.; Lorca, Graciela L.; Gonzalez, Claudio F.

    2015-01-01

    Host and commensals crosstalk, mediated by reactive oxygen species (ROS), has triggered a growing scientific interest to understand the mechanisms governing such interaction. However, the majority of the scientific studies published do not evaluate the ROS production by commensals bacteria. In this context we recently showed that Lactobacillus johnsonii N6.2, a strain of probiotic value, modulates the activity of the critical enzymes 2,3-indoleamine dioxygenase via H2O2 production. L. johnsonii N6.2 by decreasing IDO activity, is able to modify the tryptophan/kynurenine ratio in the host blood with further systemic consequences. Understanding the mechanisms of H2O2 production is critical to predict the probiotic value of these strains and to optimize bacterial biomass production in industrial processes. We performed a transcriptome analysis to identify genes differentially expressed in L. johnsonii N6.2 cells collected from cultures grown under different aeration conditions. Herein we described the biochemical characteristics of a heterodimeric FMN reductase (FRedA/B) whose in vitro activity is controlled by LjPAS protein with a typical Per-Arnst-Sim (PAS) sensor domain. Interestingly, LjPAS is fused to the FMN reductase domains in other lactobacillaceae. In L. johnsonii, LjPAS is encoded by an independent gene which expression is repressed under anaerobic conditions (>3 fold). Purified LjPAS was able to slow down the FRedA/B initial activity rate when the holoenzyme precursors (FredA, FredB, and FMN) were mixed in vitro. Altogether the results obtained suggest that LjPAS module regulates the H2O2 production helping the cells to minimize oxidative stress in response to environmental conditions. PMID:26236298

  18. H2O2 production rate in Lactobacillus johnsonii is modulated via the interplay of a heterodimeric flavin oxidoreductase with a soluble 28 Kd PAS domain containing protein.

    PubMed

    Valladares, Ricardo B; Graves, Christina; Wright, Kaitlyn; Gardner, Christopher L; Lorca, Graciela L; Gonzalez, Claudio F

    2015-01-01

    Host and commensals crosstalk, mediated by reactive oxygen species (ROS), has triggered a growing scientific interest to understand the mechanisms governing such interaction. However, the majority of the scientific studies published do not evaluate the ROS production by commensals bacteria. In this context we recently showed that Lactobacillus johnsonii N6.2, a strain of probiotic value, modulates the activity of the critical enzymes 2,3-indoleamine dioxygenase via H2O2 production. L. johnsonii N6.2 by decreasing IDO activity, is able to modify the tryptophan/kynurenine ratio in the host blood with further systemic consequences. Understanding the mechanisms of H2O2 production is critical to predict the probiotic value of these strains and to optimize bacterial biomass production in industrial processes. We performed a transcriptome analysis to identify genes differentially expressed in L. johnsonii N6.2 cells collected from cultures grown under different aeration conditions. Herein we described the biochemical characteristics of a heterodimeric FMN reductase (FRedA/B) whose in vitro activity is controlled by LjPAS protein with a typical Per-Arnst-Sim (PAS) sensor domain. Interestingly, LjPAS is fused to the FMN reductase domains in other lactobacillaceae. In L. johnsonii, LjPAS is encoded by an independent gene which expression is repressed under anaerobic conditions (>3 fold). Purified LjPAS was able to slow down the FRedA/B initial activity rate when the holoenzyme precursors (FredA, FredB, and FMN) were mixed in vitro. Altogether the results obtained suggest that LjPAS module regulates the H2O2 production helping the cells to minimize oxidative stress in response to environmental conditions.

  19. Comparative evaluation of antimicrobial efficacy of an alternative natural agent for disinfection of toothbrushes.

    PubMed

    Vignesh, R; Rekha, C Vishnu; Baghkomeh, Parisa Norouzi; Annamalai, Sankar; Sharmin, Ditto

    2017-01-01

    The aim of this study was to assess the antimicrobial potency of aqueous extract of Psidium guajava leaves in two different concentrations as a toothbrush disinfectant against three oral bacterial species. Aqueous extracts of P. guajava leaves were prepared at 20% and 30% concentrations and 0.2% chlorhexidine was used as control. The toothbrushes were equally divided into 9 groups with 10 toothbrushes per disinfectant, which were contaminated with Streptococcus mutans , Lactobacillus acidophilus , and Enterococcus faecalis . Microbial culture was done after 5 min and 3 h of decontamination. Group Ia and Ib showed that the presence of E. faecalis was observed in 8 (40%) of 20 toothbrushes. Group IIa and IIb showed a significant reduction in colony forming unit/toothbrush during 3 h evaluation. Group IIIa and IIIb showed nil growth during 3 h evaluation. Nil growth was observed with the control group for all three organisms. Statistically significant values were obtained for 5 min ( P < 0.001) and 3 h ( P < 0.001) disinfection period against L. acidophilus at two different concentrations. Aqueous extracts of guava leaves can be used as an alternative organic product for disinfection of toothbrushes.

  20. Mg2+ improves the thermotolerance of probiotic Lactobacillus rhamnosus GG, Lactobacillus casei Zhang and Lactobacillus plantarum P-8.

    PubMed

    Yang, Y; Huang, S; Wang, J; Jan, G; Jeantet, R; Chen, X D

    2017-04-01

    Food-related carbohydrates and proteins are often used as thermoprotectants for probiotic lactobacilli during industrial production and processing. However, the effect of inorganic salts is rarely reported. Magnesium is the second-most abundant cation in bacteria, and commonly found in various foods. Mg 2+ homeostasis is important in Salmonella and has been reported to play a critical role in their thermotolerance. However, the role of Mg 2+ in thermotolerance of other bacteria, in particular probiotic bacteria, still remains a hypothesis. In this study, the effect of Mg 2+ on thermotolerance of probiotic lactobacilli was investigated in three well-documented probiotic strains, Lactobacillus rhamnosus GG, Lactobacillus casei Zhang and Lactobacillus plantarum P-8, in comparison with Zn 2+ and Na + . Concentrations of Mg 2+ between 10 and 50 mmol l -1 were found to increase the bacterial survival upon heat challenge. Remarkably, Mg 2+ addition at 20 mmol l -1 led to a 100-fold higher survival of L. rhamnosus GG upon heat challenge. This preliminary study also showed that Mg 2+ shortened the heat-induced extended lag time of bacteria, which indicated the improvement in bacterial recovery from thermal injury. In order to improve the productivity and stability of live probiotics, extensive investigations have been carried out to improve thermotolerance of probiotics. However, most of these studies focused on the effects of carbohydrates, proteins or amino acids. The roles of inorganic salts in various food materials, which have rarely been reported, should be considered when incorporating probiotics into these foods. In this study, Mg 2+ was found to play a significant role in the thermotolerance of probiotic lactobacilli. A novel strategy may be available in the near future by employing magnesium salts as protective agents of probiotics during manufacturing process. © 2017 The Society for Applied Microbiology.

  1. Growth and exopolysaccharide yield of Lactobacillus delbrueckii ssp. bulgaricus DSM 20081 in batch and continuous bioreactor experiments at constant pH.

    PubMed

    Mende, Susann; Krzyzanowski, Leona; Weber, Jost; Jaros, Doris; Rohm, Harald

    2012-02-01

    Some Lactobacillus delbrueckii ssp. bulgaricus strains are able to synthesize exopolysaccharides (EPS) and are therefore highly important for the dairy industry as starter cultures. The aim of this study was to investigate the nutritional requirements for growth and EPS production of Lactobacillus delbrueckii ssp. bulgaricus DSM 20081. A medium was developed from a semi-defined medium (SDM) in which glucose was replaced by lactose and different combinations of supplements (nucleobases, vitamins, salts, sodium formate and orotic acid) were added. Constant pH batch fermentation with the modified medium resulted in an EPS yield of approximately 210 mg glucose equivalents per liter medium. This was a 10-fold increase over flask cultivation of this strain in SDM. Although not affecting cell growth, the mixture of salts enhanced the EPS synthesis. Whereas EPS production was approximately 12 mg/g dry biomass without salt supplementation, a significantly higher yield (approximately 20 mg/g dry biomass) was observed after adding the salt mixture. In continuous fermentation, a maximal EPS concentration was obtained at a dilution rate of 0.31/h (80 mg EPS/L), which corresponded to a specific EPS production of 49 mg/g dry biomass. Copyright © 2011 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  2. 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

  3. Lactobacillus plantarum isolated from kefir protects vero cells from cytotoxicity by type-II shiga toxin from Escherichia coli O157:H7.

    PubMed

    Kakisu, Emiliano; Abraham, Analía G; Farinati, Carla Tironi; Ibarra, Cristina; De Antoni, Graciela L

    2013-02-01

    Kefir is a fermented-milk beverage originating and widely consumed in the Caucasus as well as in Eastern Europe and is a source of bacteria with potential probiotic properties. Enterohaemorrhagic Escherichia coli producing Shiga toxin is commonly associated with food-transmitted diseases; the most prevalent serotype causing epidemics is Esch. coli O157:H7. The aim of this study was to evaluate the antagonism of Lactobacillus plantarum isolated from kefir against the action on Vero cells of supernatants of the Esch. coli O157:H7 strain 69160 expressing the type-II Shiga toxin (Stx2) and to study the role of the Lactobacillus cell wall in that inhibition. Spent culture supernatants of Esch. coli O157:H7 strain 69160 led to cytotoxic effects on cultured eukaryotic cells as evidenced by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-bromide-cleavage assay or by lactate-dehyrogenase release. Lb. plantarum CIDCA 83114 reduced the cytotoxic activity of Stx present in strain-69160 supernatants, and this protection was markedly higher than those of Lactobacillus kefir CIDCA 83113 and 8348 and Lb. delbrueckii subsp. bulgaricus CIDCA 333. This antagonism of cytotoxicity was mimicked by Lb. plantarum cell walls but was reduced after heating or protease treatments, thus indicating a protein or peptide as being involved in the protection mechanism. The cell surface of the lactobacilli bound the subunit B of Stx thereby decreasing the cytotoxicity. These interactions could constitute the first step in preventing the damage induced by Esch. coli O157:H7 supernatants, thus representing a valuable means of potentially mitigating the noxious effects of this food pathogen.

  4. Mutational Analysis of the Stability of the H2A and H2B Histone Monomers

    PubMed Central

    Stump, Matthew R.; Gloss, Lisa M.

    2008-01-01

    The eukaryotic histone heterodimer H2A-H2B folds through an obligatory dimeric intermediate that forms in a nearly diffusion-limited association reaction in the stopped-flow dead time. It is unclear whether there is partial folding of the isolated monomers before association. To address the possible contributions of structure in the monomers to the rapid association, we characterized H2A and H2B monomers in the absence of their heterodimeric partner. By far-UV circular dichroism, the H2A and H2B monomers are 15% and 31% helical, respectively—significantly less than observed in X-ray crystal structures. Acrylamide quenching of the intrinsic Tyr fluorescence was indicative of tertiary structure. The H2A and H2B monomers exhibit free energies of unfolding of 2.5 and 2.9 kcal mol−1, respectively; at 10 μM, the sum of the stability of the monomers is ~60% of the stability of the native dimer. The helical content, stability and m values indicate that H2B has a more stable, compact structure than H2A. The monomer m values are larger than expected for the extended histone fold motif, suggesting that the monomers adopt an overly-collapsed structure. Stopped-flow refolding—initiated from urea-denatured monomers or the partially folded monomers populated at low denaturant concentrations—yielded essentially identical rates, indicating that monomer folding is productive in the rapid association and folding of the heterodimer. A series of Ala and Gly mutations were introduced into H2A and H2B to probe the importance of helix propensity on the structure and stability of the monomers. The mutational studies show that the central α-helix of the histone fold, which makes extensive inter-monomer contacts, is structured in H2B but only partially folded in H2A. PMID:18976667

  5. Insights into glycogen metabolism in Lactobacillus acidophilus: impact on carbohydrate metabolism, stress tolerance and gut retention.

    PubMed

    Goh, Yong Jun; Klaenhammer, Todd R

    2014-11-20

    In prokaryotic species equipped with glycogen metabolism machinery, the co-regulation of glycogen biosynthesis and degradation has been associated with the synthesis of energy storage compounds and various crucial physiological functions, including global cellular processes such as carbon and nitrogen metabolism, energy sensing and production, stress response and cell-cell communication. In addition, the glycogen metabolic pathway was proposed to serve as a carbon capacitor that regulates downstream carbon fluxes, and in some microorganisms the ability to synthesize intracellular glycogen has been implicated in host persistence. Among lactobacilli, complete glycogen metabolic pathway genes are present only in select species predominantly associated with mammalian hosts or natural environments. This observation highlights the potential involvement of glycogen biosynthesis in probiotic activities and persistence of intestinal lactobacilli in the human gastrointestinal tract. In this review, we summarize recent findings on (i) the presence and potential ecological distribution of glycogen metabolic pathways among lactobacilli, (ii) influence of carbon substrates and growth phases on glycogen metabolic gene expression and glycogen accumulation in L. acidophilus, and (iii) the involvement of glycogen metabolism on growth, sugar utilization and bile tolerance. Our present in vivo studies established the significance of glycogen biosynthesis on the competitive retention of L. acidophilus in the mouse intestinal tract, demonstrating for the first time that the ability to synthesize intracellular glycogen contributes to gut fitness and retention among probiotic microorganisms.

  6. Suppression of Rotational Twins in Epitaxial B 12P 2 on 4H-SiC

    DOE PAGES

    Frye, C. D.; Saw, C. K.; Padavala, Balabalaji; ...

    2017-12-22

    B 12P2 was grown epitaxially on (0001) 4H-SiC using two different substrate miscuts: a standard 4° miscut toward the [more » $$11\\bar{20}$$] and a custom miscut 4° toward the [$$1\\bar{10}0$$]. Epitaxy on substrates miscut to the [$$11\\bar{20}$$] resulted in highly twinned B 12P 2 films with a rotational twin density of approximately 70% twin orientation I and 30% twin orientation II. In contrast, epitaxy on substrates tilted toward the [$$1\\bar{10}0$$] produced films of >99% twin orientation I. A H 2 etch model is used to explain the 4H-SiC surface morphology for each miscut prior to epitaxy and demonstrate how the surface steps influence the nucleation of B 12P 2 twin orientations. Surface steps on substrates miscut to the [$$11\\bar{20}$$] tend to be zig-zagged with steps rotated 60° from one another producing B 12P 2 crystals that nucleate in orientations rotated by 60°, hence forming rotationally twinned films. In conclusion, steps on substrates tilted to the [$$1\\bar{10}0$$] tend to be parallel resulting in crystallographically aligned B 12P 2 nucleation.« less

  7. Suppression of Rotational Twins in Epitaxial B 12P 2 on 4H-SiC

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Frye, C. D.; Saw, C. K.; Padavala, Balabalaji

    B 12P2 was grown epitaxially on (0001) 4H-SiC using two different substrate miscuts: a standard 4° miscut toward the [more » $$11\\bar{20}$$] and a custom miscut 4° toward the [$$1\\bar{10}0$$]. Epitaxy on substrates miscut to the [$$11\\bar{20}$$] resulted in highly twinned B 12P 2 films with a rotational twin density of approximately 70% twin orientation I and 30% twin orientation II. In contrast, epitaxy on substrates tilted toward the [$$1\\bar{10}0$$] produced films of >99% twin orientation I. A H 2 etch model is used to explain the 4H-SiC surface morphology for each miscut prior to epitaxy and demonstrate how the surface steps influence the nucleation of B 12P 2 twin orientations. Surface steps on substrates miscut to the [$$11\\bar{20}$$] tend to be zig-zagged with steps rotated 60° from one another producing B 12P 2 crystals that nucleate in orientations rotated by 60°, hence forming rotationally twinned films. In conclusion, steps on substrates tilted to the [$$1\\bar{10}0$$] tend to be parallel resulting in crystallographically aligned B 12P 2 nucleation.« less

  8. 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

  9. Lactobacillus acidophilus counteracts enteropathogenic E. coli-induced inhibition of butyrate uptake in intestinal epithelial cells

    PubMed Central

    Kumar, Anoop; Alrefai, Waddah A.; Dudeja, Pradeep K.

    2015-01-01

    Butyrate, a key short-chain fatty acid metabolite of colonic luminal bacterial action on dietary fiber, serves as a primary fuel for the colonocytes, ameliorates mucosal inflammation, and stimulates NaCl absorption. Absorption of butyrate into the colonocytes is essential for these intracellular effects. Monocarboxylate transporter 1 (MCT1) plays a major role in colonic luminal butyrate absorption. Previous studies (Tan J, McKenzie C, Potamitis M, Thorburn AN, Mackay CR, Macia L. Adv Immunol 121: 91–119, 2014.) showed decreased MCT1 expression and function in intestinal inflammation. We have previously shown (Borthakur A, Gill RK, Hodges K, Ramaswamy K, Hecht G, Dudeja PK. Am J Physiol Gastrointest Liver Physiol 290: G30–G35, 2006.) impaired butyrate absorption in human intestinal epithelial Caco-2 cells due to decreased MCT1 level at the apical cell surface following enteropathogenic E. coli (EPEC) infection. Current studies, therefore, examined the potential role of probiotic Lactobacilli in stimulating MCT1-mediated butyrate uptake and counteracting EPEC inhibition of MCT1 function. Of the five species of Lactobacilli, short-term (3 h) treatment with L. acidophilus (LA) significantly increased MCT1-mediated butyrate uptake in Caco-2 cells. Heat-killed LA was ineffective, whereas the conditioned culture supernatant of LA (LA-CS) was equally effective in stimulating MCT1 function, indicating that the effects are mediated by LA-secreted soluble factor(s). Furthermore, LA-CS increased apical membrane levels of MCT1 protein via decreasing its basal endocytosis, suggesting that LA-CS stimulation of butyrate uptake could be secondary to increased levels of MCT1 on the apical cell surface. LA-CS also attenuated EPEC inhibition of butyrate uptake and EPEC-mediated endocytosis of MCT1. Our studies highlight distinct role of specific LA-secreted molecules in modulating colonic butyrate absorption. PMID:26272259

  10. 77 FR 10037 - Temporary Non-Agricultural Employment of H-2B Aliens in the United States

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-02-21

    ... Administration 20 CFR Part 655 Wage and Hour Division 29 CFR Part 503 Temporary Non-Agricultural Employment of H... Employment of H-2B Aliens in the United States AGENCY: Employment and Training Administration, and Wage and... nonimmigrant worker in H-2B status. We have also created new regulations to provide for increased worker...

  11. In vitro evaluation of the viability of vaginal cells (VK2/E6E7) and probiotic Lactobacillus species in lemon juice.

    PubMed

    Anukam, Kingsley C; Reid, Gregor

    2009-03-01

    Women, especially in developing countries, most often bear the brunt of HIV infections. The continued lack of viable vaccines and microbicides has made some women resort to using natural products such as lemon or lime juice to avoid infection. Few in vitro studies have been done on the effect of lemon juice on vaginal cells and lactobacilli that constitute the major microbiota in healthy women. The objective of the present study was to evaluate in vitro the effect of lemon juice on the viability of vaginal cells (VK2/E6E7) and vaginal Lactobacillus species. Vaginal cells were exposed to different concentrations (0-30%) of lemon juice at pH 2.3 and 4.5 for 10 min. Viability was determined by staining the cells with propidium iodide and analysing them by flow cytometry. Lactobacillus organisms were dispensed into microplates with vaginally defined medium + peptone (VDMP) containing different concentrations of lemon juice ranging from 0 to 100%. Lemon juice at pH 2.3 had a significant (P = 0.03) toxic effect on the vaginal cell line used. At 30% concentration, the vaginal cells were practically non-viable, typified by a 95% loss of viability, whereas at pH 4.5 there was only 5% cell loss. Lemon juice had varying growth inhibitory effects on the Lactobacillus species tested. At pH 4.5 and using 10-30% lemon juice, there was a stimulatory growth effect on certain Lactobacillus species. Lemon juice (20-30%) at pH 2.3 was highly toxic to VK2/E6E7 cells, and at pH 4.5 there was no significant effect on the viability of the cells within 10 min. Lemon juice above 10% at pH 2.3 was found to be detrimental to the growth of vaginal lactobacilli. Although lemon juice may be useful in other applications, its use in the vaginal region should be discouraged.

  12. The nido-osmaboranes [2,2,2-(CO)(PPh(3))(2)-nido-2-OsB(5)H(9)] and [6,6,6-(CO)(PPh(3))(2)-nido-6-OsB(9)H(13)].

    PubMed

    Bould, J; Kennedy, J D; Thomas, R L; Rath, N P; Barton, L

    2001-11-01

    The structural characterization of the osmahexaborane 2-carbonyl-2,2-bis(triphenylphosphine)-nido-2-osmahexaborane(9), [Os(B(5)H(9))(C(18)H(15)P)(2)(CO)], (I), a metallaborane analogue of B(6)H(10), confirms the structure proposed from NMR spectroscopy. The structure of the osmadecaborane 6-carbonyl-6,6-bis(triphenylphosphine)-nido-6-osmadecaborane(13), [Os(B(9)H(13))(C(18)H(15)P)(2)(CO)], (IV), is similarly confirmed. The short basal B-B distance of 1.652 (8) A in (I), not bridged by an H atom, mirrors that in the parent hexaborane(10) [1.626 (4) A].

  13. [The range of antagonistic effects of Lactobacillus bacterial strains on etiologic agents of bacterial vaginosis].

    PubMed

    Strus, M; Malinowska, M

    1999-01-01

    Bacterial vaginosis is caused by uncontrolled sequential overgrowth of some anaerobic bacteria: Gardnerella vaginalis, Prevotella bivia, Bacteroides spp., Peptostreptococcus spp., Mobiluncus sp. usually occurring in stable numbers in the bacterial flora of healthy women. On the other hand, different species of bacteria belonging to the genus Lactobacillus, most frequently L. plantarum, L. rhamnosus and L. acidophilus, form a group of aerobic bacteria dominating in the same environment. The diversity and density of their populations depend on the age and health conditions. Thanks to their antagonistic and adherence properties bacteria of the genus Lactobacillus can maintain a positive balance role in this ecosystem. The aim of this study was to assess the antagonistic properties of Lactobacillus strains isolated from the vagina of healthy women against most common agents of bacterial vaginosis. It was found that nearly all of the tested Lactobacillus strains exerted distinct antagonistic activity against anaerobic bacteria: Gardnerella vaginalis, Prevotella bivia and Peptostreptococcus anaerobius and quite a number also against Gram-negative rods, while only some of them were able to inhibit Gram-positive aerobic cocci as Enterococcus faecalis or Staphylococcus aureus.

  14. Development of an integration mutagenesis system in Lactobacillus gasseri.

    PubMed

    Selle, Kurt; Goh, Yong Jun; O'Flaherty, Sarah; Klaenhammer, Todd R

    2014-01-01

    Lactobacillus gasseri ATCC 33323 is a member of the acidophilus-complex group, microbes of human origin with significant potential for impacting human health based on niche-specific traits. In order to facilitate functional analysis of this important species, a upp-based counterselective chromosomal integration system was established and employed for targeting the lipoteichoic acid (LTA) synthesis gene, ltaS, in L. gasseri ATCC 33323. The ltaS gene encodes a phosphoglycerol transferase responsible for building the glycerol chain of LTA. No isogenic mutant bearing the deletion genotype was recovered, but an integration knockout mutant was generated with insertion inactivation at the ltaS locus. The ltaS deficient derivative exhibited an altered cellular morphology and significantly reduced ability to adhere to Caco-2 intestinal cell monolayers, relative to the wild-type parent strain.

  15. Saccharomyces boulardii expresses neuraminidase activity selective for α2,3-linked sialic acid that decreases Helicobacter pylori adhesion to host cells.

    PubMed

    Sakarya, Serhan; Gunay, Necati

    2014-10-01

    Helicobacter pylori is a major causative agent of gastritis and peptic ulcer disease and is an established risk factor for gastric malignancy. Antibiotic combination therapy can eradicate H. pylori. As these same regimens can evoke adverse effects and resistance, new alternative therapies or adjunctive treatments are needed. A probiotic approach may provide a novel strategy for H. pylori treatment. In the current study, two probiotic bacteria, Lactobacillus acidophilus and Lactobacillus reuteri, and a probiotic yeast, Saccharomyces boulardii, were evaluated for their ability to influence H. pylori viability, adherence to gastric and duodenal cells, as well as the effect of S. boulardii on cell surface expression of sialic acid. Our results indicate that S. boulardii contains neuraminidase activity selective for α(2-3)-linked sialic acid. This neuraminidase activity removes surface α(2-3)-linked sialic acid, the ligand for the sialic acid-binding H. pylori adhesin, which in turn, inhibits H. pylori adherence to duodenal epithelial cells. © 2014 APMIS. Published by John Wiley & Sons Ltd.

  16. High-level expression of recombinant beta-galactosidases in Lactobacillus plantarum and Lactobacillus sakei using a Sakacin P-based expression system.

    PubMed

    Halbmayr, Elisabeth; Mathiesen, Geir; Nguyen, Thu-Ha; Maischberger, Thomas; Peterbauer, Clemens K; Eijsink, Vincent G H; Haltrich, Dietmar

    2008-06-25

    This work presents the cloning and expression of the genes encoding heterodimeric beta-galactosidases from Lactobacillus reuteri L103, Lactobacillus acidophilus R22, Lactobacillus plantarum WCFS1, and Lactobacillus sakei Lb790. These enzymes consist of two subunits of approximately 73 and 35 kDa, which are encoded by two overlapping genes, lacL and lacM, respectively. We have cloned these genes into the lactobacillal expression vectors pSIP403 and pSIP409, which are based on the sakacin P operon of L. sakei ( Sørvig et al. Microbiology 2005, 151, 2439- 2449 ), and expressed them in the host strains L. plantarum WCFS1 and L. sakei Lb790. Results varied considerably, ranging from 2.23 to 61.1 U/mg of beta-galactosidase activity, depending on the origin of the lacLM genes, the host strain, and the expression vector used. Highest expression levels were obtained in a laboratory cultivation of L. plantarum WCFS1 harboring the plasmid pEH3R containing the lacLM gene from L. reuteri L103. These cultivations yielded approximately 23 000 U of beta-galactosidase activity per liter, corresponding to the formation of roughly 100 mg of recombinant protein per liter of fermentation medium, and beta-galactosidase levels amounted to 55% of the total intracellular protein of the host organism. To further verify the suitability of this expression system, recombinant beta-galactosidase from L. reuteri was purified to apparent homogeneity. The properties of the purified enzyme were essentially identical with the properties of purified native beta-galactosidase from L. reuteri L103. The presented results lead the way to efficient overproduction of beta-galactosidase in a food-grade expression system, which is of high interest for applications in food industry.

  17. Gardnerella vaginalis and Lactobacillus sp in liquid-based cervical samples in healthy and disturbed vaginal flora using cultivation-independent methods.

    PubMed

    Klomp, Johanna M; Verbruggen, Banut-Sabine M; Korporaal, Hans; Boon, Mathilde E; de Jong, Pauline; Kramer, Gerco C; van Haaften, Maarten; Heintz, A Peter M

    2008-05-01

    Our objective was to determine the morphotype of the adherent bacteria in liquid-based cytology (LBC) in smears with healthy and disturbed vaginal flora. And to use PCR technology on the same fixed cell sample to establish DNA patterns of the 16S RNA genes of the bacteria in the sample. Thirty samples were randomly selected from a large group of cervical cell samples suspended in a commercial coagulant fixative "(BoonFix)." PCR was used to amplify DNA of five bacterial species: Lactobacillus acidophilus, Lactobacillus crispatus, Lactobacillus jensenii, Gardnerella vaginalis, and Mycoplasma hominis. The LBC slides were then analyzed by light microscopy to estimate bacterial adhesion. DNA of lactobacilli was detected in all cell samples. Seventeen smears showed colonization with Gardnerella vaginalis (range 2.6 x 10(2)-3.0 x 10(5) bacteria/mul BoonFix sample). Two cases were identified as dysbacteriotic with high DNA values for Gardnerella vaginalis and low values for Lactobacillus crispatus. The sample with the highest concentration for Gardnerella vaginalis showed an unequivocal Gardnerella infection. This study indicates that the adherence pattern of a disturbed flora in liquid-based cervical samples can be identified unequivocally, and that these samples are suitable for quantitative PCR analysis. This cultivation independent method reveals a strong inverse relationship between Gardnerella vaginalis and Lactobacillus crispatus in dysbacteriosis and unequivocal Gardnerella infection.

  18. Lactobacillus micheneri sp. nov., Lactobacillus timberlakei sp. nov. and Lactobacillus quenuiae sp. nov., lactic acid bacteria isolated from wild bees and flowers.

    PubMed

    McFrederick, Quinn S; Vuong, Hoang Q; Rothman, Jason A

    2018-06-01

    Gram-stain-positive, rod-shaped, non-spore forming bacteria have been isolated from flowers and the guts of adult wild bees in the families Megachilidae and Halictidae. Phylogenetic analysis of the 16S rRNA gene indicated that these bacteria belong to the genus Lactobacillus, and are most closely related to the honey-bee associated bacteria Lactobacillus kunkeei (97.0 % sequence similarity) and Lactobacillus apinorum (97.0 % sequence similarity). Phylogenetic analyses of 16S rRNA genes and six single-copy protein coding genes, in situ and in silico DNA-DNA hybridization, and fatty-acid profiling differentiates the newly isolated bacteria as three novel Lactobacillus species: Lactobacillus micheneri sp. nov. with the type strain Hlig3 T (=DSM 104126 T ,=NRRL B-65473 T ), Lactobacillus timberlakei with the type strain HV_12 T (=DSM 104128 T ,=NRRL B-65472 T ), and Lactobacillus quenuiae sp. nov. with the type strain HV_6 T (=DSM 104127 T ,=NRRL B-65474 T ).

  19. 77 FR 28764 - Temporary Non-agricultural Employment of H-2B Aliens in the United States

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-05-16

    ... DEPARTMENT OF LABOR Employment and Training Administration 20 CFR Part 655 RIN 1205-AB58 Temporary Non-agricultural Employment of H-2B Aliens in the United States AGENCY: Employment and Training... Temporary Non-agricultural Employment of H-2B Aliens in the United States, published February 21, 2012 (the...

  20. Probiotic Lactobacillus Strains Stimulate the Inflammatory Response and Activate Human Macrophages.

    PubMed

    Rocha-Ramírez, L M; Pérez-Solano, R A; Castañón-Alonso, S L; Moreno Guerrero, S S; Ramírez Pacheco, A; García Garibay, M; Eslava, C

    2017-01-01

    Lactobacilli have been shown to promote health functions. In this study, we analyzed the mechanism by which four different strains of probiotics affected innate immunity, such as regulation of ROS, cytokines, phagocytosis, bactericidal activity, signaling by NF- κ B pp65, and TLR2 activation. The production of ROS was dependent on the concentration and species of Lactobacillus . The results obtained from the tested strains ( Lactobacillus rhamnosus GG, L. rhamnosus KLSD, L. helveticus IMAU70129, and L. casei IMAU60214) showed that strains induced early proinflammatory cytokines such as IL-8,TNF- α , IL-12p70, and IL-6. However, IL-1 β expression was induced only by L. helveticus and L. casei strains (after 24 h stimulation). Phagocytosis and bactericidal activity of macrophages against various pathogens, such as S. aureus , S. typhimurium , and E. coli , were increased by pretreatment with Lactobacillus . The nuclear translocation NF- κ B pp65 and TLR2-dependent signaling were also increased by treatment with the probiotics. Taken together, the experiments demonstrate that probiotic strains of Lactobacillus exert early immunostimulatory effects that may be directly linked to the initial inflammation of the response of human macrophages.

  1. Probiotic Lactobacillus Strains Stimulate the Inflammatory Response and Activate Human Macrophages

    PubMed Central

    Moreno Guerrero, S. S.; Ramírez Pacheco, A.; García Garibay, M.; Eslava, C.

    2017-01-01

    Lactobacilli have been shown to promote health functions. In this study, we analyzed the mechanism by which four different strains of probiotics affected innate immunity, such as regulation of ROS, cytokines, phagocytosis, bactericidal activity, signaling by NF-κB pp65, and TLR2 activation. The production of ROS was dependent on the concentration and species of Lactobacillus. The results obtained from the tested strains (Lactobacillus rhamnosus GG, L. rhamnosus KLSD, L. helveticus IMAU70129, and L. casei IMAU60214) showed that strains induced early proinflammatory cytokines such as IL-8,TNF-α, IL-12p70, and IL-6. However, IL-1β expression was induced only by L. helveticus and L. casei strains (after 24 h stimulation). Phagocytosis and bactericidal activity of macrophages against various pathogens, such as S. aureus, S. typhimurium, and E. coli, were increased by pretreatment with Lactobacillus. The nuclear translocation NF-κB pp65 and TLR2-dependent signaling were also increased by treatment with the probiotics. Taken together, the experiments demonstrate that probiotic strains of Lactobacillus exert early immunostimulatory effects that may be directly linked to the initial inflammation of the response of human macrophages. PMID:28758133

  2. Production of Succinic Acid from Citric Acid and Related Acids by Lactobacillus Strains

    PubMed Central

    Kaneuchi, Choji; Seki, Masako; Komagata, Kazuo

    1988-01-01

    A number of Lactobacillus strains produced succinic acid in de Man-Rogosa-Sharpe broth to various extents. Among 86 fresh isolates from fermented cane molasses in Thailand, 30 strains (35%) produced succinic acid; namely, 23 of 39 Lactobacillus reuteri strains, 6 of 18 L. cellobiosus strains, and 1 of 6 unidentified strains. All of 10 L. casei subsp. casei strains, 5 L. casei subsp. rhamnosus strains, 6 L. mali strains, and 2 L. buchneri strains did not produce succinic acid. Among 58 known strains including 48 type strains of different Lactobacillus species, the strains of L. acidophilus, L. crispatus, L. jensenii, and L. parvus produced succinic acid to the same extent as the most active fresh isolates, and those of L. alimentarius, L. collinoides, L. farciminis, L. fructivorans (1 of 2 strains tested), L. malefermentans, and L. reuteri were also positive, to lesser extents. Diammonium citrate in de Man-Rogosa-Sharpe broth was determined as a precursor of the succinic acid produced. Production rates were about 70% on a molar basis with two fresh strains tested. Succinic acid was also produced from fumaric and malic acids but not from dl-isocitric, α-ketoglutaric, and pyruvic acids. The present study is considered to provide the first evidence on the production of succinic acid, an important flavoring substance in dairy products and fermented beverages, from citrate by lactobacilli. PMID:16347795

  3. Production of succinic Acid from citric Acid and related acids by lactobacillus strains.

    PubMed

    Kaneuchi, C; Seki, M; Komagata, K

    1988-12-01

    A number of Lactobacillus strains produced succinic acid in de Man-Rogosa-Sharpe broth to various extents. Among 86 fresh isolates from fermented cane molasses in Thailand, 30 strains (35%) produced succinic acid; namely, 23 of 39 Lactobacillus reuteri strains, 6 of 18 L. cellobiosus strains, and 1 of 6 unidentified strains. All of 10 L. casei subsp. casei strains, 5 L. casei subsp. rhamnosus strains, 6 L. mali strains, and 2 L. buchneri strains did not produce succinic acid. Among 58 known strains including 48 type strains of different Lactobacillus species, the strains of L. acidophilus, L. crispatus, L. jensenii, and L. parvus produced succinic acid to the same extent as the most active fresh isolates, and those of L. alimentarius, L. collinoides, L. farciminis, L. fructivorans (1 of 2 strains tested), L. malefermentans, and L. reuteri were also positive, to lesser extents. Diammonium citrate in de Man-Rogosa-Sharpe broth was determined as a precursor of the succinic acid produced. Production rates were about 70% on a molar basis with two fresh strains tested. Succinic acid was also produced from fumaric and malic acids but not from dl-isocitric, alpha-ketoglutaric, and pyruvic acids. The present study is considered to provide the first evidence on the production of succinic acid, an important flavoring substance in dairy products and fermented beverages, from citrate by lactobacilli.

  4. Unveiling of novel regio-selective fatty acid double bond hydratases from Lactobacillus acidophilus involved in the selective oxyfunctionalization of mono- and di-hydroxy fatty acids.

    PubMed

    Kim, Kyoung-Rok; Oh, Hye-Jin; Park, Chul-Soon; Hong, Seung-Hye; Park, Ji-Young; Oh, Deok-Kun

    2015-11-01

    The aim of this study is the first time demonstration of cis-12 regio-selective linoleate double-bond hydratase. Hydroxylation of fatty acids, abundant feedstock in nature, is an emerging alternative route for many petroleum replaceable products thorough hydroxy fatty acids, carboxylic acids, and lactones. However, chemical route for selective hydroxylation is still quite challenging owing to low selectivity and many environmental concerns. Hydroxylation of fatty acids by hydroxy fatty acid forming enzymes is an important route for selective biocatalytic oxyfunctionalization of fatty acids. Therefore, novel fatty acid hydroxylation enzymes should be discovered. The two hydratase genes of Lactobacillus acidophilus were identified by genomic analysis, and the expressed two recombinant hydratases were identified as cis-9 and cis-12 double-bond selective linoleate hydratases by in vitro functional validation, including the identification of products and the determination of regio-selectivity, substrate specificity, and kinetic parameters. The two different linoleate hydratases were the involved enzymes in the 10,13-dihydroxyoctadecanoic acid biosynthesis. Linoleate 13-hydratase (LHT-13) selectively converted 10 mM linoleic acid to 13S-hydroxy-9(Z)-octadecenoic acid with high titer (8.1 mM) and yield (81%). Our study will expand knowledge for microbial fatty acid-hydroxylation enzymes and facilitate the designed production of the regio-selective hydroxy fatty acids for useful chemicals from polyunsaturated fatty acid feedstocks. © 2015 Wiley Periodicals, Inc.

  5. Assessment of the effect of probiotic curd consumption on salivary pH and streptococcus mutans counts.

    PubMed

    Sudhir, R; Praveen, P; Anantharaj, A; Venkataraghavan, Karthik

    2012-07-01

    Antimicrobial methods of controlling dental caries that include probiotic agents can play a valuable role in establishing caries control in children at moderate to high risk for developing dental caries. Several studies have demonstrated the beneficial effects of use of various Probiotic products including curd. The objective of this study was to compare the effect of short-term consumption of probiotic curd containing Lactobacillus acidophilus and normal curd on salivary Streptococcus Mutans counts, as well as salivary pH. Forty, caries-free, 10-12 years old children were selected and randomly allocated to two groups. Test Group consisted of 20 children who consumed 200ml of probiotic curd daily for 30 days. Control Group consisted of 20 children who were given 200ml of regular curd for 30 days. Salivary pH and salivary Streptococcus Mutans counts were recorded at baseline and after 30 days and statistically compared using the Student's t-test. Consumption of probiotic curd resulted in a statistically significant reduction in S. Mutans colony counts (P<0.001) as compared to regular curd. However, there was a slight reduction in pH (P>0.05) in both the groups. Short-term consumption of probiotic curds can reduce oral S. Mutans counts. However, this caused a slight reduction in salivary pH.

  6. Production of Fermented Kale Juices with Lactobacillus Strains and Nutritional Composition

    PubMed Central

    Kim, Seong Yeong

    2017-01-01

    Fermented kale juices using four types of lactobacilli were produced in the present study. After 48 h of fermentation time, viable cell counts of all ferments reached an above 109 CFU/mL. The viability of the ferments after cold storage in the refrigerator for 4 weeks showed 108 CFU/mL in all ferments. Among four types of fermented kale juices, the ferment of Lactobacillus acidophilus IFO 3025 indicated a good nutritional composition, including neutral sugar (1,909.76 μg/mL), reducing sugar (564.00 μg/mL, P<0.05), and protein contents (160.06 μg/mL, P<0.05). The results of mineral composition analysis had the highest potassium value in all ferments (854.16~895.07 μg/mL), particularly in the ferment of Lactobacillus brevis FSB-1 (P<0.001), which is necessary to sustain osmotic pressure, prevention of high blood pressure, and protein synthesis. Moreover, calcium, phosphorus, and magnesium contents related to bone health were generally sufficient in all ferments. Consequently, in this study, fermented kale juices may be suggested as a healthy fermented beverage with essential nutrients. However, the acceptability of the fermented kale juice to the Korean taste should be further investigated with a trained taste panel to determine whether inoculated fermentation could be an option for the consumers. PMID:29043222

  7. Probiotic Dahi containing Lactobacillus acidophilus and Bifidobacterium bifidum modulates the formation of aberrant crypt foci, mucin-depleted foci, and cell proliferation on 1,2-dimethylhydrazine-induced colorectal carcinogenesis in Wistar rats.

    PubMed

    Mohania, Dheeraj; Kansal, Vinod K; Kruzliak, Peter; Kumari, Archana

    2014-08-01

    Aberrant crypt foci (ACF) and mucin-depleted foci (MDF) are pre-neoplastic lesions identified in the colon of carcinogen-treated rodents and in humans at high risk for colon cancer. The present study was carried out to divulge the protective potential of the probiotic Dahi containing Lactobacillus acidophilus LaVK2 and Bifidobacterium bifidum BbVK3 alone or in combination with piroxicam (PXC) on the development of early biomarkers of colorectal carcinogenesis in male Wistar rats administered 1,2-dimethylhydrazine (DMH). DMH was injected subcutaneously at the rate of 40 mg/kg body weight per animal twice a week for 2 weeks. A total of 120 male Wistar rats were randomly allocated to five groups, each group having 24 animals. The rats were fed with buffalo milk or probiotic supplement (20 grams) alone or as an adjunct with PXC in addition to a basal diet ad libitum for 32 weeks. Group I was offered buffalo milk (BM) and served as the control group. Group II was administered DMH along with BM and served as the DMH-control group; group III was administered BM-DMH-PXC, in which besides administering BM-DMH, PXC was also offered. Group IV was offered probiotic LaBb Dahi and DMH, and group V was offered both probiotic LaBb Dahi and PXC along with DMH. The rats were euthanized at the 8(th), 16(th), and 32(nd) week of the experiment and examined for development of ACF, aberrant crypts per ACF (AC/ACF), mucin-depleted foci (MDF), large MDF, and proliferating cell nuclear antigen (PCNA) labeling index. Administration of DMH in rats induced pre-neoplastic lesions (ACF and MDF) and increased the PCNA index in colorectal tissue. A significant (p<0.05) reduction in the number of ACF, AC/ACF, MDF, large MDF, and PCNA labeling index were observed in the probiotic LaBb Dahi group compared with the DMH control group. Feeding rats with LaBb Dahi or treatment with PXC diminished the initiation and progression of DMH-induced pre-neoplastic lesions and the PCNA index, and treatment with

  8. Synthesis, characterization, antimicrobial and enzymatic activity of 4b,9b-dihydroxy-7,8-dihydro-4bH-indeno[1,2-b]benzofuran-9,10(6H,9bH)-dione

    NASA Astrophysics Data System (ADS)

    Mehdi, Sayed Hasan; Hashim, Rokiah; Ghalib, Raza Murad; Fátima C. Guedes da Silva, M.; Sulaiman, Othman; Rahman, Syed Ziaur; Murugaiyah, Vikneswaran; Marimuthu, Mani Maran

    2011-12-01

    The crystal structure of the title compound, 4b,9b-dihydroxy-7,8-dihydro-4bH-indeno[1,2-b]benzofuran-9,10(6H,9bH)-dione has been determined by single crystal X-ray diffraction. It crystallizes in the monoclinic space group P2 1/c with Z = 4. The FTIR as well as the 1H and 13C NMR spectra of the compound were also recorded and briefly discussed. The compound showed potential antimicrobial activity comparable to that of clinically used antimicrobial agents against selected microorganisms. It has selective and moderate inhibitory activity on butyryl cholinesterase enzyme and could serve as potential lead compound for synthesis of more bioactive derivatives.

  9. Molecular mechanisms and in vitro antioxidant effects of Lactobacillus plantarum MA2.

    PubMed

    Tang, Wei; Xing, Zhuqing; Li, Chao; Wang, Jinju; Wang, Yanping

    2017-04-15

    Lactobacillus plantarum MA2 was isolated from Chinese traditional Tibetan kefir grains. The antioxidant activities in vitro of this strain were evaluated extensively. The results showed that L. plantarum MA2 can tolerate hydrogen peroxide up to 2.0mM, and its fermentate (fermented supernatant, intact cell and cell-free extract) had strong reducing capacities, lipid peroxidation inhibition capacities, Fe 2+ -chelating abilities, as well as various free radical scavenging capacities. Additionally, both the fermented supernatant and cell homogenate exhibited glutathione peroxidase activity and superoxide dismutase activity. In order to investigate the antioxidant mechanism of L. plantarum MA2 at the molecular level, eight antioxidant-related genes were identified, and further analyzed. Three groups of genes cat, gshR and npx, were found up-regulated under H 2 O 2 challenge. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. A Computational Investigation of the Oxidative Deboronation of BoroGlycine, H2N–CH2B(OH)2, Using H2O and H2O2

    PubMed Central

    Larkin, Joseph D.; Markham, George D.; Milkevitch, Matt; Brooks, Bernard R.; Bock, Charles W.

    2014-01-01

    We report results from a computational investigation of the oxidative deboronation of BoroGlycine, H2N–CH2B(OH)2, using H2O and H2O2 as the reactive oxygen species (ROS) to yield aminomethanol, H2N–CH2–OH; these results complement our study on the protodeboronation of BoroGlycine to produce methylamine, H2N–CH3 (Larkin et al. J. Phys. Chem. A, 111, 6489–6500, 2007). Second-order Møller-Plesset (MP2) perturbation theory with Dunning-Woon correlation-consistent (cc) basis sets were used for the calculations with comparisons made to results from Density Functional Theory (DFT) at the PBE1PBE/6-311++G(d,p)(cc-pVDZ) levels. The effects of a bulk aqueous environment were also incorporated into the calculations employing PCM and CPCM methodology. Using H2O as the ROS, the reaction H2O + H2N–CH2B(OH)2H2N–CH2–OH + H–B(OH)2 was calculated to be endothermic, the value of ΔH2980 was +12.0 kcal/mol at the MP2(FC)/cc-pVTZ computational level in vacuo and +13.7 kcal/mol in PCM aqueous media; the corresponding value for the activation barrier, ΔH‡, was +94.3 kcal/mol relative to the separated reactants in vacuo and +89.9 kcal/mol in PCM aqueous media. In contrast, the reaction H2O2 + H2N–CH2B(OH)2H2N–CH2–OH + B(OH)3 was calculated to be highly exothermic with a ΔH2980 value of −100.9 kcal/mol at the MP2(FC)/cc-pVTZ computational level in vacuo and −99.6 kcal/mol in CPCM aqueous media; the highest-energy transition state for the multi-step process associated with this reaction involved the rearrangement of H2N–CH2B(OH)(OOH) to H2N–CH2–O–B(OH)2 with a ΔH‡ value of +23.2 kcal/mol in vacuo relative to the separated reactants. These computational results for BoroGlycine are in accord with the experimental observations for the deboronation of the FDA approved anti-cancer drug Bortezomib (Velcade™, PS-341) where it was found to be the principle deactivation pathway. (Labutti et al. Chem. Res. Toxicol., 19, 539–546

  11. Inelastic scattering matrix elements for the nonadiabatic collision B(2P1/2)+H2(1Sigmag+,j)<-->B(2P3/2)+H2(1Sigmag+,j').

    PubMed

    Weeks, David E; Niday, Thomas A; Yang, Sang H

    2006-10-28

    Inelastic scattering matrix elements for the nonadiabatic collision B(2P1/2)+H2(1Sigmag+,j)<-->B(2P3/2)+H2(1Sigmag+,j') are calculated using the time dependent channel packet method (CPM). The calculation employs 1 2A', 2 2A', and 1 2A" adiabatic electronic potential energy surfaces determined by numerical computation at the multireference configuration-interaction level [M. H. Alexander, J. Chem. Phys. 99, 6041 (1993)]. The 1 2A' and 2 2A', adiabatic electronic potential energy surfaces are transformed to yield diabatic electronic potential energy surfaces that, when combined with the total B+H2 rotational kinetic energy, yield a set of effective potential energy surfaces [M. H. Alexander et al., J. Chem. Phys. 103, 7956 (1995)]. Within the framework of the CPM, the number of effective potential energy surfaces used for the scattering matrix calculation is then determined by the size of the angular momentum basis used as a representation. Twenty basis vectors are employed for these calculations, and the corresponding effective potential energy surfaces are identified in the asymptotic limit by the H2 rotor quantum numbers j=0, 2, 4, 6 and B electronic states 2Pja, ja=1/2, 3/2. Scattering matrix elements are obtained from the Fourier transform of the correlation function between channel packets evolving in time on these effective potential energy surfaces. For these calculations the H2 bond length is constrained to a constant value of req=1.402 a.u. and state to state scattering matrix elements corresponding to a total angular momentum of J=1/2 are discussed for j=0<-->j'=0,2,4 and 2P1/2<-->2P1/2, 2P3/2 over a range of total energy between 0.0 and 0.01 a.u.

  12. IS30-related transposon mediated insertional inactivation of bile salt hydrolase (bsh1) gene of Lactobacillus plantarum strain Lp20.

    PubMed

    Kumar, Rajesh; Grover, Sunita; Kaushik, Jai K; Batish, Virender Kumar

    2014-01-01

    Lactobacillus plantarum is a flexible and versatile microorganism that inhabits a variety of niches, and its genome may express up to four bsh genes to maximize its survival in the mammalian gut. However, the ecological significance of multiple bsh genes in L. plantarum is still not clearly understood. Hence, this study demonstrated the disruption of bile salt hydrolase (bsh1) gene due to the insertion of a transposable element in L. plantarum Lp20 - a wild strain of human fecal origin. Surprisingly, L. plantarum strain Lp20 produced a ∼2.0 kb bsh1 amplicon against the normal size (∼1.0 kb) bsh1 amplicon of Bsh(+)L. plantarum Lp21. Strain Lp20 exhibited minimal Bsh activity in spite of having intact bsh2, bsh3 and bsh4 genes in its genome and hence had a Bsh(-) phenotype. Cloning and sequence characterization of Lp20 bsh1 gene predicted four individual open reading frames (ORFs) within this region. BLAST analysis of ORF1 and ORF2 revealed significant sequence similarity to the L. plantarum bsh1 gene while ORF3 and ORF4 showed high sequence homology to IS30-family transposases. Since, IS30-related transposon element was inserted within Lp20 bsh1 gene in reverse orientation (3'-5'), it introduced several stop codons and disrupted the protein reading frames of both Bsh1 and transposase. Inverted terminal repeats (GGCAGATTG) of transposon, mediated its insertion at 255-263 nt and 1301-1309 nt positions of Lp20 bsh1 gene. In conclusion, insertion of IS30 related-transposon within the bsh1 gene sequence of L. plantarum strain Lp20 demolished the integrity and functionality of Bsh1 enzyme. Additionally, this transposon DNA sequence remains active among various Lactobacillus spp. and hence harbors the potential to be explored in the development of efficient insertion mutagenesis system. Copyright © 2013 Elsevier GmbH. All rights reserved.

  13. Lactobacillus brevis CD2 inhibits Prevotella melaninogenica biofilm.

    PubMed

    Vuotto, C; Barbanti, F; Mastrantonio, P; Donelli, G

    2014-10-01

    To evaluate the ability of the probiotic strain Lactobacillus brevis CD2 to inhibit the opportunistic anaerobe Prevotella melaninogenica (PM1), a well-known causative agent of periodontitis. The inhibitory effect of Lactobacillus CD2 on Prevotella PM1 biofilm was assessed both by exposing the anaerobe to the supernatant of the probiotic strain and by growing the two strains to obtain single or mixed biofilms. The inhibitory effect of CD2 on PM1 was also checked by the agar overlay method. The development of PM1 biofilm was strongly affected (56% decrease in OD value) by the CD2 supernatant after 96 h. A dose-dependent biofilm reduction was also observed at 1/10 and 1/100 dilutions of supernatant. Confocal microscopy on the mixed biofilms revealed the ability of CD2 to prevail on PM1, greatly reducing the biofilm of the latter. It has been hypothesized a multifactorial nature of the inhibition mechanism, the strong adherence ability of CD2 strain together with the released metabolites presumably contributing to the reduction in the PM1 biofilm detected by confocal microscopy. © 2013 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  14. 30 CFR 57.22238 - Actions at 2.0 percent methane (I-B, II-B, V-B, and VI mines).

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... 30 Mineral Resources 1 2011-07-01 2011-07-01 false Actions at 2.0 percent methane (I-B, II-B, V-B, and VI mines). 57.22238 Section 57.22238 Mineral Resources MINE SAFETY AND HEALTH ADMINISTRATION... Actions at 2.0 percent methane (I-B, II-B, V-B, and VI mines). If methane reaches 2.0 percent in the mine...

  15. 30 CFR 57.22238 - Actions at 2.0 percent methane (I-B, II-B, V-B, and VI mines).

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... 30 Mineral Resources 1 2010-07-01 2010-07-01 false Actions at 2.0 percent methane (I-B, II-B, V-B... AND NONMETAL MINES Safety Standards for Methane in Metal and Nonmetal Mines Ventilation § 57.22238 Actions at 2.0 percent methane (I-B, II-B, V-B, and VI mines). If methane reaches 2.0 percent in the mine...

  16. Hydride CVD Hetero-epitaxy of B 12P 2 on 4H-SiC

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Frye, C. D.; Saw, C. K.; Padavala, Balabalaji

    Icosahedral boron phosphide (B 12P 2) is a wide bandgap semiconductor (3.35 eV) that has been reported to “self-heal” from high-energy electron bombardment, making it attractive for potential use in radioisotope batteries, radiation detection, or in electronics in high radiation environments. Our study focused on improving B 12P 2 hetero-epitaxial films by growing on 4H-SiC substrates over the temperature range of 1250–1450 °C using B 2H 6 and PH 3 precursors in a H 2 carrier gas. Furthermore, XRD scans and Laue transmission photographs revealed that the epitaxial relationship was (0001)<11more » $$\\bar{2}$$0> B12P2|| (0001)<11$$\\bar{2}$$0> 4H-SiC. The film morphology and crystallinity were investigated as a function of growth temperature and growth time. At 1250 °C, films tended to form rough, polycrystalline layers, but at 1300 and 1350 °C, films were continuous and comparatively smooth (R RMS≤7 nm). At 1400 or 1450 °C, the films grew in islands that coalesced as the films became thicker. Using XRD rocking curves to evaluate the crystal quality, 1300 °C was the optimum growth temperature tested. Finally, at 1300 °C, the rocking curve FWHM decreased with increasing film thickness from 1494 arcsec for a 1.1 μm thick film to 954 arcsec for a 2.7 µm thick film, suggesting a reduction in defects with thickness.« less

  17. Hydride CVD Hetero-epitaxy of B 12P 2 on 4H-SiC

    DOE PAGES

    Frye, C. D.; Saw, C. K.; Padavala, Balabalaji; ...

    2016-11-27

    Icosahedral boron phosphide (B 12P 2) is a wide bandgap semiconductor (3.35 eV) that has been reported to “self-heal” from high-energy electron bombardment, making it attractive for potential use in radioisotope batteries, radiation detection, or in electronics in high radiation environments. Our study focused on improving B 12P 2 hetero-epitaxial films by growing on 4H-SiC substrates over the temperature range of 1250–1450 °C using B 2H 6 and PH 3 precursors in a H 2 carrier gas. Furthermore, XRD scans and Laue transmission photographs revealed that the epitaxial relationship was (0001)<11more » $$\\bar{2}$$0> B12P2|| (0001)<11$$\\bar{2}$$0> 4H-SiC. The film morphology and crystallinity were investigated as a function of growth temperature and growth time. At 1250 °C, films tended to form rough, polycrystalline layers, but at 1300 and 1350 °C, films were continuous and comparatively smooth (R RMS≤7 nm). At 1400 or 1450 °C, the films grew in islands that coalesced as the films became thicker. Using XRD rocking curves to evaluate the crystal quality, 1300 °C was the optimum growth temperature tested. Finally, at 1300 °C, the rocking curve FWHM decreased with increasing film thickness from 1494 arcsec for a 1.1 μm thick film to 954 arcsec for a 2.7 µm thick film, suggesting a reduction in defects with thickness.« less

  18. Maximum-biomass prediction of homofermentative Lactobacillus.

    PubMed

    Cui, Shumao; Zhao, Jianxin; Liu, Xiaoming; Chen, Yong Q; Zhang, Hao; Chen, Wei

    2016-07-01

    Fed-batch and pH-controlled cultures have been widely used for industrial production of probiotics. The aim of this study was to systematically investigate the relationship between the maximum biomass of different homofermentative Lactobacillus and lactate accumulation, and to develop a prediction equation for the maximum biomass concentration in such cultures. The accumulation of the end products and the depletion of nutrients by various strains were evaluated. In addition, the minimum inhibitory concentrations (MICs) of acid anions for various strains at pH 7.0 were examined. The lactate concentration at the point of complete inhibition was not significantly different from the MIC of lactate for all of the strains, although the inhibition mechanism of lactate and acetate on Lactobacillus rhamnosus was different from the other strains which were inhibited by the osmotic pressure caused by acid anions at pH 7.0. When the lactate concentration accumulated to the MIC, the strains stopped growing. The maximum biomass was closely related to the biomass yield per unit of lactate produced (YX/P) and the MIC (C) of lactate for different homofermentative Lactobacillus. Based on the experimental data obtained using different homofermentative Lactobacillus, a prediction equation was established as follows: Xmax - X0 = (0.59 ± 0.02)·YX/P·C. Copyright © 2016. Published by Elsevier B.V.

  19. Optimization of growth and bacteriocin production by Lactobacillus sakei subsp. sakei2a.

    PubMed

    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 2(4) 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).

  20. Lactobacillus species: taxonomic complexity and controversial susceptibilities.

    PubMed

    Goldstein, Ellie J C; Tyrrell, Kerin L; Citron, Diane M

    2015-05-15

    The genus Lactobacillus is a taxonomically complex and is composed of over 170 species that cannot be easily differentiated phenotypically and often require molecular identification. Although they are part of the normal human gastrointestinal and vaginal flora, they can also be occasional human pathogens. They are extensively used in a variety of commercial products including probiotics. Their antimicrobial susceptibilities are poorly defined in part because of their taxonomic complexity and are compounded by the different methods recommended by Clinical Laboratory Standards Institute and International Dairy Foundation. Their use as probiotics for prevention of Clostridium difficile infection is prevalent among consumers worldwide but raises the question of will the use of any concurrent antibiotic effect their ability to survive. Lactobacillus species are generally acid resistant and are able to survive ingestion. They are generally resistant to metronidazole, aminoglycosides and ciprofloxacin with L. acidophilus being susceptible to penicillin and vancomycin, whereas L. rhamnosus and L. casei are resistant to metronidazole and vancomycin. © The Author 2015. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  1. Antimicrobial potential for the combination of bovine lactoferrin or its hydrolysate with lactoferrin-resistant probiotics against foodborne pathogens.

    PubMed

    Chen, P-W; Jheng, T T; Shyu, C-L; Mao, F C

    2013-03-01

    Previous reports have shown that several probiotic strains can resist the antibacterial activity of bovine lactoferrin (bLf), but the results are inconsistent. Moreover, a portion of orally administered apo-bLf is digested in vivo by pepsin to yield bLf hydrolysate, which produces stronger antibacterial activity than that observed with apo-bLf. However, whether bLf hydrolysate affects the growth of probiotic strains is unclear. Therefore, various probiotic strains in Taiwan were collected and evaluated for activity against apo-bLf and bLf hydrolysate in vitro. Thirteen probiotic strains were evaluated, and the growth of Lactobacillus acidophilus ATCC 4356, Lactobacillus salivarius ATCC 11741, Lactobacillus rhamnosus ATCC 53103, Bifidobacterium longum ATCC 15707, and Bifidobacterium lactis BCRC 17394 were inhibited by both apo-bLf and bLf hydrolysate. The growth of 8 strains were not affected by apo-bLf and bLf hydrolysate, including L. rhamnosus ATCC 7469, Lactobacillus reuteri ATCC 23272, Lactobacillus fermentum ATCC 11739, Lactobacillus coryniformis ATCC 25602, L. acidophilus BCRC 14065, Bifidobacterium infantis ATCC 15697, Bifidobacterium bifidum ATCC 29521, and Pediococcus acidilactici ATCC 8081. However, apo-bLf and its hydrolysate inhibited the growth of foodborne pathogens, including Escherichia coli, Salmonella typhimurium, Staphylococcus aureus, and Enterococcus faecalis. Moreover, the supernatants produced by L. fermentum, B. lactis, and B. longum inhibited the growth of most pathogens. Importantly, a combination of apo-bLf or bLf hydrolysate with the supernatants of cultures of the organisms described above showed synergistic or partially synergistic effects against the growth of most of the selected pathogens. In conclusion, several probiotic strains are resistant to apo-bLf and bLf hydrolysate, warranting clinical studies to evaluate the antimicrobial potential for the combination of apo-bLf or its hydrolysate with specific probiotics. Copyright

  2. Draft Genome Sequence of Lactobacillus delbrueckii subsp. bulgaricus LBB.B5.

    PubMed

    Urshev, Zoltan; Hajo, Karima; Lenoci, Leonardo; Bron, Peter A; Dijkstra, Annereinou; Alkema, Wynand; Wels, Michiel; Siezen, Roland J; Minkova, Svetlana; van Hijum, Sacha A F T

    2016-10-06

    Lactobacillus delbrueckii subsp. bulgaricus LBB.B5 originates from homemade Bulgarian yogurt and was selected for its ability to form a strong association with Streptococcus thermophilus The genome sequence will facilitate elucidating the genetic background behind the contribution of LBB.B5 to the taste and aroma of yogurt and its exceptional protocooperation with S. thermophilus. Copyright © 2016 Urshev et al.

  3. Diselenophosphate-Induced Conversion of an Achiral [Cu 20 H 11 {S 2 P(O i Pr) 2 } 9 ] into a Chiral [Cu 20 H 11 {Se 2 P(O i Pr) 2 } 9 ] Polyhydrido Nanocluster

    DOE PAGES

    Dhayal, Rajendra S.; Liao, Jian-Hong; Wang, Xiaoping; ...

    2015-11-09

    A polyhydrido copper nanocluster, [Cu 20H 11{Se 2P(OiPr) 2} 9] (2 H), which exhibits an intrinsically chiral inorganic core of C-3 symmetry, was synthesized from achiral [Cu 20H 11{S 2P(OiPr) 2} 9] (1(H)) of C-3h symmetry by a ligand-exchange method. Likewise, the structure has a distorted cuboctahedral Cu-13 core, two triangular faces of which are capped along the C-3 axis, one by a Cu-6 cupola and the other by a single Cu atom. The Cu-20 framework is further stabilized by 9 diselenophosphate and 11 hydride ligands. The number of hydride, phosphorus, and selenium resonances and their splitting patterns in multinuclearmore » NMR spectra of 2(H) indicate that the chiral Cu20H11 core retains its C-3 symmetry in solution. Moreover, the 11 hydride ligands were located by neutron diffraction experiments and shown to be capping (3)-H and interstitial (5)-H ligands (in square-pyramidal and trigonal-bipyramidal cavities), as supported by DFT calculations on [Cu 20H 11(Se 2PH 2) 9] (2 H') as a simplified model.« less

  4. 21 CFR 73.3123 - 6-Ethoxy-2-(6-ethoxy-3-oxobenzo[b]thien-2(3H)-ylidene) benzo[b]thiophen-3 (2H)-one.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... 21 Food and Drugs 1 2013-04-01 2013-04-01 false 6-Ethoxy-2-(6-ethoxy-3-oxobenzo[b]thien-2(3H)-ylidene) benzo[b]thiophen-3 (2H)-one. 73.3123 Section 73.3123 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL LISTING OF COLOR ADDITIVES EXEMPT FROM CERTIFICATION Medical...

  5. 21 CFR 73.3123 - 6-Ethoxy-2-(6-ethoxy-3-oxobenzo[b]thien-2(3H)-ylidene) benzo[b]thiophen-3 (2H)-one.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 21 Food and Drugs 1 2012-04-01 2012-04-01 false 6-Ethoxy-2-(6-ethoxy-3-oxobenzo[b]thien-2(3H)-ylidene) benzo[b]thiophen-3 (2H)-one. 73.3123 Section 73.3123 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL LISTING OF COLOR ADDITIVES EXEMPT FROM CERTIFICATION Medical...

  6. 21 CFR 73.3123 - 6-Ethoxy-2-(6-ethoxy-3-oxobenzo[b]thien-2(3H)-ylidene) benzo[b]thiophen-3 (2H)-one.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... 21 Food and Drugs 1 2011-04-01 2011-04-01 false 6-Ethoxy-2-(6-ethoxy-3-oxobenzo[b]thien-2(3H)-ylidene) benzo[b]thiophen-3 (2H)-one. 73.3123 Section 73.3123 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL LISTING OF COLOR ADDITIVES EXEMPT FROM CERTIFICATION Medical...

  7. 21 CFR 73.3123 - 6-Ethoxy-2-(6-ethoxy-3-oxobenzo[b]thien-2(3H)-ylidene) benzo[b]thiophen-3 (2H)-one.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 21 Food and Drugs 1 2014-04-01 2014-04-01 false 6-Ethoxy-2-(6-ethoxy-3-oxobenzo[b]thien-2(3H)-ylidene) benzo[b]thiophen-3 (2H)-one. 73.3123 Section 73.3123 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL LISTING OF COLOR ADDITIVES EXEMPT FROM CERTIFICATION Medical...

  8. 21 CFR 73.3123 - 6-Ethoxy-2-(6-ethoxy-3-oxobenzo[b]thien-2(3H)-ylidene) benzo[b]thiophen-3 (2H)-one.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... 21 Food and Drugs 1 2010-04-01 2010-04-01 false 6-Ethoxy-2-(6-ethoxy-3-oxobenzo[b]thien-2(3H)-ylidene) benzo[b]thiophen-3 (2H)-one. 73.3123 Section 73.3123 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL LISTING OF COLOR ADDITIVES EXEMPT FROM CERTIFICATION Medical...

  9. Detoxification of Aflatoxin B1 by Antifungal Compounds from Lactobacillus brevis and Lactobacillus paracasei, Isolated from Dairy Products.

    PubMed

    Gomaa, Eman Zakaria; Abdelall, Manal Farouk; El-Mahdy, Omima Mohammed

    2018-06-01

    Aflatoxins are a large group of highly toxic, mutagenic, and carcinogenic mycotoxins produced by specific species of fungi. Potential contamination of food commodities by these compounds causes extensive damage that lead to great economic losses. This study explored the potential use of antifungal compounds, produced by Lactobacillus brevis and Lactobacillus paracasei, for growth inhibition and subsequent aflatoxin B1 production from select strains of Aspergillus flavus and Aspergillus parasiticus. Lactobacilli strains were isolated from traditional Egyptian dairy products, whereas fungal strains were isolated from infected cereal seeds. There were noticeable decreases in mycelium biomass and aflatoxin production as well. L. brevis exhibited the highest reduction of aflatoxin B1 production by A. flavus and A. parasiticus, 96.31 and 90.43%, respectively. The concentrations of amino acids of the antifungal compound produced by L. brevis were significantly higher than that produced by L. paracasei. Asparagine, glutamine, glycine, alanine, and leucine were the most concentrated amino acids for both strains. The antifungal compounds produced by L. brevis and L. paracasei were active in a wide range of pH, heat stable and inactivated by proteolytic enzymes (protease K and trypsin A). The expression of Omt-A gene that involved in the later step of aflatoxin production was evaluated by real-time PCR. There was a vigorous reduction at transcriptional level of Omt-A gene observed in A. flavus that is treated by L. brevis and L. paracasei (80 and 70%, respectively). However, the reduction of Omt-A gene observed in A. parasiticus that is treated by L. brevis and L. paracasei was 64.5 and 52%, respectively. Treating maize seeds with antifungal compounds exhibited great efficiency in controlling fungal infection and increasing seed germination. The results confirmed that lactic acid bacteria are a promising strategy to control food contamination of fermented food and dairy

  10. Novel antibacterial polypeptide produced by Lactobacillus paracasei strain NRRL B-50314

    USDA-ARS?s Scientific Manuscript database

    This study reports the production and characterization of a novel antibacterial polypeptide, designated as laparaxin, which is secreted by Lactobacillus paracasei NRRL B-50314. The crude laparaxin has antibacterial activity against a range of Gram-positive bacteria including the following: lactic a...

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

    USDA-ARS?s Scientific Manuscript database

    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...

  12. Protection of chickens against H9N2 avian influenza virus challenge with recombinant Lactobacillus plantarum expressing conserved antigens.

    PubMed

    Yang, Wen-Tao; Yang, Gui-Lian; Shi, Shao-Hua; Liu, Yu-Ying; Huang, Hai-Bin; Jiang, Yan-Long; Wang, Jian-Zhong; Shi, Chun-Wei; Jing, Yu-Bei; Wang, Chun-Feng

    2017-06-01

    Avian influenza virus (AIV) is spreading worldwide and is a serious threat to the health of poultry and humans. In many countries, low pathogenic AIVs, such as H9N2, have become an enormous economic burden on the commercial poultry industry because they cause mild respiratory disease and decrease egg production. A recombinant Lactobacillus plantarum NC8 strain expressing NP-M1-DCpep from H9N2 AIV has been studied in a mouse model. However, it remains unknown whether this L. plantarum strain can induce an immune response and provide protection against H9N2 AIV in chickens. In this study, chickens that were orally vaccinated with NC8-pSIP409-NP-M1-DCpep exhibited significantly increased T cell-mediated immune responses and mucosal sIgA and IgG levels, which provided protection against H9N2 AIV challenge. More importantly, compared with oral administration of NC8-pSIP409-NP-M1-DCpep, intranasal administration induced stronger immune responses and provided effective protection against challenge with the H9N2 virus by reducing body weight loss, lung virus titers, and throat pathology. Taken together, these findings suggest that L. plantarum expressing NP-M1-DCpep has potential as a vaccine to combat H9N2 AIV infection.

  13. Potentially probiotic Lactobacillus strains from traditional Kurdish cheese.

    PubMed

    Hashemi, Seyed Mohammad Bagher; Shahidi, Fakhri; Mortazavi, Seyed Ali; Milani, Elnaz; Eshaghi, Zarrin

    2014-03-01

    In this study, the probiotic potential of Lactobacillus strains isolated from traditional Kurdish cheese was investigated. The Lactobacillus strains were examined for resistance to gastric acidity and bile toxicity, antimicrobial activities, autoaggregation, coaggregation, hydrophobicity, adhesion to Caco-2 cells, and antibiotic susceptibility. The results showed that all strains tested tolerate acid gastric conditions (pH 2.0 and 3.0), and all of them were bile resistant (at 0.3 and 1% concentration). Although no antibacterial activity was detected in vitro assay for the treated (neutralized to pH 6.5 and treated with catalase) cell-free culture supernatant (CFCS) of strains, untreated CFCS showed strong antagonistic activity against two known pathogens bacteria. All strains exhibited a strong autoaggregating phenotype and manifested a high degree of coaggregation with pathogens. On the other hand, majority of studied strains were found sensitive to different antibiotics, such as ampicillin, penicillin, ciprofloxacin, chloramphenicol, erythromycin, rifampicin, and tetracycline, and were resistant to vancomycin and streptomycin. Finally, isolated strains showed good hydrophobicity and adherence to Caco-2 cell line, so they could be exploited for food manufacture.

  14. 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

  15. H2B ubiquitination: Conserved molecular mechanism, diverse physiologic functions of the E3 ligase during meiosis.

    PubMed

    Wang, Liying; Cao, Chunwei; Wang, Fang; Zhao, Jianguo; Li, Wei

    2017-09-03

    RNF20/Bre1 mediated H2B ubiquitination (H2Bub) has various physiologic functions. Recently, we found that H2Bub participates in meiotic recombination by promoting chromatin relaxation during meiosis. We then analyzed the phylogenetic relationships among the E3 ligase for H2Bub, its E2 Rad6 and their partner WW domain-containing adaptor with a coiled-coil (WAC) or Lge1, and found that the molecular mechanism underlying H2Bub is evolutionarily conserved from yeast to mammals. However, RNF20 has diverse physiologic functions in different organisms, which might be caused by the evolutionary divergency of their domain/motif architectures. In the current extra view, we not only elucidate the evolutionarily conserved molecular mechanism underlying H2Bub, but also discuss the diverse physiologic functions of RNF20 during meiosis.

  16. Encapsulation of probiotic bacteria in lamb rennet paste: effects on the quality of Pecorino cheese.

    PubMed

    Santillo, A; Albenzio, M; Bevilacqua, A; Corbo, M R; Sevi, A

    2012-07-01

    Lamb rennet pastes containing encapsulated Lactobacillus acidophilus and a mix of Bifidobacterium longum and Bifidobacterium lactis were produced for Pecorino cheese manufacture from Gentile di Puglia ewe milk. Cheeses were denoted as RP cheese when made with traditional rennet paste, RP-L cheese when made with rennet paste containing L. acidophilus culture, and RP-B cheese when made with rennet paste containing a mix of B. lactis and B. longum. Biochemical features of Pecorino cheese were studied at 1, 15, 30, 60, and 120 d of cheese ripening. The effect of encapsulation and bead addition to rennet acted on a different way on the viability of probiotic. Lactobacillus acidophilus retained its viability for 4 to 5 d and then showed a fast reduction; on the other hand, B. longum and B. lactis experienced kinetics characterized by an initial death slope, followed by a tail effect due to acquired resistance. At 1 d of ripening, the levels of L. acidophilus and bifidobacteria in cheese were the lowest, and then increased, reaching the highest levels after 30 d; such cell loads were maintained throughout the ripening for L. acidophilus, whereas bifidobacteria experienced a decrease of about 1 log cfu/g at the end of ripening. Enzymatic activities and biochemical features of cheeses were influenced by the type of rennet used for cheesemaking. Greater enzymatic activity was recorded in RP-L and RP-B cheese due to the presence of probiotic bacteria released from alginate beads. A positive correlation was found between enzymatic activities and water-soluble nitrogen and proteose-peptone in RP-B and RP-L cheeses; water-soluble nitrogen and proteose-peptone were the highest in RP-B. Principal component analysis distinguished RP-L from the other cheeses on the basis of the conjugated linoleic acid content, which was higher in the RP-L due to the ability of L. acidophilus to produce conjugated linoleic acid in the cheese matrix. Copyright © 2012 American Dairy Science

  17. Gene-specific characterization of human histone H2B by electron capture dissociation.

    PubMed

    Siuti, Nertila; Roth, Michael J; Mizzen, Craig A; Kelleher, Neil L; Pesavento, James J

    2006-02-01

    The basis set of protein forms expressed by human cells from the H2B gene family was determined by Top Down Mass Spectrometry. Using Electron Capture Dissociation for MS/MS of H2B isoforms, direct evidence for the expression of unmodified H2B.Q, H2B.A, H2B.K/T, H2B.J, H2B.E, H2B.B, H2B.F, and monoacetylated H2B.A was obtained from asynchronous HeLa cells. H2B.A was the most abundant form, with the overall expression profile not changing significantly in cells arrested in mitosis by colchicine or during mid-S, mid-G2, G2/M, and mid-G1 phases of the cell cycle. Modest hyperacetylation of H2B family members was observed after sodium butyrate treatment.

  18. 78 FR 24047 - Wage Methodology for the Temporary Non-Agricultural Employment H-2B Program, Part 2

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-04-24

    ... Employment and Training Administration 20 CFR Part 655 RIN 1205-AB69 Wage Methodology for the Temporary Non-Agricultural Employment H- 2B Program, Part 2 AGENCY: Employment and Training Administration, Labor; U.S. Citizenship and Immigration Services, DHS. ACTION: Interim final rule; request for comments. SUMMARY: The...

  19. EphB4-targeted imaging with antibody h131, h131-F(ab′)2 and h131-Fab

    PubMed Central

    Li, Dan; Liu, Shuanglong; Liu, Ren; Zhou, Yue; Park, Ryan; Naga, Kranthi; Krasnoperov, Valery; Gill, Parkash S.; Li, Zibo; Shan, Hong; Conti, Peter S.

    2013-01-01

    Accumulating evidence suggests that overexpression of the tyrosine kinase receptor EphB4, a mediator of vascular development, is a novel target for tumor diagnosis, prognosis and therapy. Noninvasive imaging of EphB4 expression could therefore be valuable for evaluating disease course and therapeutic efficacy at the earliest stages of anti-EphB4 treatment. In this study, we systematically investigated the use of anti-EphB4 antibody h131 (150 kD) and its fragments (h131-F(ab′)2, 110 kD; h131-Fab, 50 kD) for near-infrared fluorescence (NIRF) imaging of EphB4 expression in vivo. h131-F(ab′)2 and h131-Fab were produced through pepsin and papain digestion of h131 respectively, whose purity was confirmed by FPLC and SDS-PAGE. After conjugation with Cy5.5, in vivo characteristics of h131, h131-F(ab′)2 and h131-Fab were evaluated in EphB4-positive HT29 tumor model. Although h131-Cy5.5 demonstrated highest tumor uptake among these probes, its optimal tumor uptake level was obtained at 2 d post injection (p.i.). For h131-Fab-Cy5.5, maximum tumor uptake was achieved at 4 h p.i.. However, no significant difference was observed between h131-Fab-Cy5.5 and hIgG-Fab-Cy5.5, indicating the tumor accumulation was mainly caused by passive targeting. In contrast, h131-F(ab′)2-Cy5.5 demonstrated prominent tumor uptake at 6 h p.i. The target specificity was confirmed by hIgG-F(ab′)2-Cy5.5 control and immunofluorescent staining. Collectively, h131-F(ab′)2 exhibited prominent and specific tumor uptake at early time points, which suggests it is a promising agent for EphB4-targeted imaging. PMID:24147882

  20. Field Trials of Attenuated Salmonella Typhi Live Oral Vaccine TY21A in Liquid and Enteric-Coated Capsule Formulations in Santiago, Chile

    DTIC Science & Technology

    1990-06-01

    inactive piacebo, children in the control group received viable Lactobacillus acidophilus because some experimental data suggest that L. acidophilus may...was the enteric-coating used to make the capsules acid-resistant. Such capsules, each containing 1-3 x 109 viable vaccine (or Lactobacillus ) organisms...formulation of vaccine (or of the Lactobacillus control preparation) consisted of two aluminum foil packets, one containing lyophilized vaccine (or

  1. Impact of inulin and okara on Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 viability in a fermented soy product and probiotic survival under in vitro simulated gastrointestinal conditions.

    PubMed

    Bedani, Raquel; Rossi, Elizeu Antonio; Isay Saad, Susana Marta

    2013-06-01

    The effect of inulin and/or okara flour on Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 viability in a fermented soy product (FSP) and on probiotic survival under in vitro simulated gastrointestinal conditions were investigated throughout 28 days of storage at 4 °C. Employing a 2(2) design, four FSP trials were produced from soymilk fermented with ABT-4 culture (La-5, Bb-12, and Streptococcus thermophilus): FSP (control); FSP-I (with inulin, 3 g/100 mL of soymilk); FSP-O (with okara, 5 g/100 mL); FSP-IO (with inulin + okara, ratio 3:5 g/100 mL). Probiotic viabilities ranged from 8 to 9 log cfu/g during the 28 days of storage, and inulin and/or okara flour did not affect the viability of La-5 and Bb-12. Bb-12 resistance to the artificial gastrointestinal juices was higher than for La-5, since the Bb-12 and La-5 populations decreased approximately 0.6 log cfu/g and 3.8 log cfu/g, respectively, throughout storage period. Even though the protective effect of inulin and/or okara flour on probiotic microorganisms was not significant, when compared to a fresh culture, the FSP matrix improved Bb-12 survival on day 1 of storage and may be considered a good vehicle for Bb-12 and could play an important role in probiotic protection against gastrointestinal juices. Copyright © 2013 Elsevier Ltd. All rights reserved.

  2. In situ synthesis of Fe-based alloy clad coatings containing TiB2-TiN-(h-BN)

    NASA Astrophysics Data System (ADS)

    Jiang, Shao-qun; Wang, Gang; Ren, Qing-wen; Yang, Chuan-duo; Wang, Ze-hua; Zhou, Ze-hua

    2015-06-01

    Fe-based alloy coatings containing TiB2-TiN-(h-BN) were synthesized in situ on Q235 steel substrates by a plasma cladding process using the powders of Fe901 alloy, Ti, and h-BN as raw materials. The effects of Ti/h-BN mass ratio on interfacial bonds between the coating and substrate along with the microstructures and microhardnesses of the coatings were investigated. The results show that the Ti/h-BN mass ratio is a vital factor in the formation of the coatings. Free h-BN can be introduced into the coatings by adding an excess amount of h-BN into the precursor. Decreases in the Ti/h-BN mass ratio improve the microstructural uniformity and compactness and enhance the interfacial bonds of the coatings. At a Ti/h-BN mass ratio of 10/20, the coating is free of cracks and micropores, and mainly consists of Fe-Cr, Fe3B, TiB2, TiN, Ti2N, TiB, FeN, FeB, Fe2B, and h-BN phases. Its average microhardness in the zone between 0.1-2.8 mm from the coating surface is about Hv0.2 551.5.

  3. Short belt-like Ca 2 B 2 O 5 ·H 2 O nanostructures: Hydrothermal formation, FT-IR, thermal decomposition, and optical properties

    NASA Astrophysics Data System (ADS)

    Zhu, Wancheng; Zhang, Xiao; Wang, Xiaoli; Zhang, Heng; Zhang, Qiang; Xiang, Lan

    2011-10-01

    Uniform high crystallinity short belt-like Ca 2B 2O 5·H 2O nanostructures (nanobelts) were facilely synthesized through a room temperature coprecipitation of CaCl 2, H 3BO 3, and NaOH solutions, followed by a mild hydrothermal treatment (180 °C, 12.0 h). By a preferential growth parallel to the (1 0 0) planes, Ca 2B 2O 5·H 2O nanobelts with a length of 1-5 μm, a width of 100-400 nm, and a thickness of 55-90 nm were obtained. The calcination of the nanobelts at 500 °C for 2.0 h led to short Ca 2B 2O 5 nanobelts with well preserved 1D morphology. Calcination at 800 °C led to a mixture of Ca 2B 2O 5 and Ca 3B 2O 6. The products were with belt-like and quasi-polyhedron morphology, while they turned into pore-free micro-rod like and polyhedron morphology when the calcination was taken in the presence of NaCl. NaCl assisted high temperature calcination at 900 °C promoted the formation of Ca 3B 2O 6 in the products. When dispersed in deionized water or absolute ethanol, the Ca 2B 2O 5·H 2O nanobelts and Ca 2B 2O 5 nanobelts showed good transparency from the ultraviolet to the visible region. The as-synthesized Ca 2B 2O 5·H 2O and Ca 2B 2O 5 nanobelts can be employed as novel metal borate nanomaterials for further potential applications in the area of glass fibers, antiwear additive, ceramic coatings, and so on.

  4. Interaction of soy isoflavones and their main metabolites with hOATP2B1 transporter.

    PubMed

    Navrátilová, Lucie; Applová, Lenka; Horký, Pavel; Mladěnka, Přemysl; Pávek, Petr; Trejtnar, František

    2018-06-22

    Membrane organic anion-transporting polypeptides (OATPs) are responsible for the drug transmembrane transport within the human body. The function of OATP2B1 transporter can be inhibited by various natural compounds. Despite increased research interest in soya as a part of human diet, the effect of its active components to interact with hOATP2B1 has not been elucidated in a complex extent. This in vitro study examined the inhibitory effect of main soy isoflavones (daidzin, daidzein, genistin, genistein, glycitin, glycitein, biochanin A, formononetin) and their metabolites formed in vivo (S-equol, O-desmethylangolensin) towards human OATP2B1 transporter. MDCKII cells overexpressing hOATP2B1 were employed to determine quantitative inhibitory parameters of the tested compounds and to analyze mechanism/s of the inhibitory interaction. The study showed that aglycones of soy isoflavones and the main biologically active metabolite S-equol were able to significantly inhibit hOATP2B1-mediated transport. The K i values for most of aglycones range from 1 to 20 μM. In contrast, glucosides did not exhibit significant inhibitory effect. The kinetic analysis did not indicate a uniform type of inhibition towards the hOATP2B1 although predominant mechanism of inhibition seemed to be competitive. These findings may suggest that tested soy isoflavones and their metabolites might affect transport of xenobiotics including drugs across tissue barriers via hOATP2B1.

  5. Mixotrophic and autotrophic growth of Thiobacillus acidophilus on glucose and thiosulfate

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Pronk, J.T.; Meulenberg, R.; van den Berg, D.J.C.

    1990-11-01

    Mixotrophic growth of the facultatively autotrophic acidophile Thiobacillus acidophilus on mixtures of glucose and thiosulfate or tetrathionate was studied in substrate-limited chemostat cultures. Growth yields in mixotrophic cultures were higher than the sum of the heterotrophic and autographic growth yields. Pulse experiments with thiosulfate indicated that tetrathionate is an intermediate during thiosulfate oxidation by cell suspensions of T. acidophilus. From mixotrophic growth studied, the energetic value of thiosulfate and tetrathionate redox equivalents was estimated to be 50% of that of redox equivalents derived from glucose oxidation. Ribulose 1,5-bisphosphate carboxylase (RuBPCase) activities in cell extracts and rates of sulfur compound oxidationmore » by cell suspensions increased with increasing thiosulfate/glucose ratios in the influent medium of the mixotrophic cultures. Significant RuBPCase and sulfur compound-oxidizing activities were detected in heterotrophically grown T. acidophilus. Polyhedral inclusion bodies (carboxysomes) could be observed at low frequencies in thin sections of cells grown in heterotrophic, glucose-limited chemostat cultures. Highest RuBPCase activities and carboxysome abundancy were observed in cells from autotrophic, CO{sub 2}-limited chemostat cultures. The maximum growth rate at which thiosulfate was still completely oxidized was increased when glucose was utilized simultaneously. This, together with the fact that even during heterotrophic growth the organism exhibited significant activities of enzymes involved in autotrophic metabolism, indicates that T. acidophilus is well adapted to a mixotrophic lifestyle. In this respect, T. acidophilus may have a competitive advantage over autotrophic acidophiles with respect to the sulfur compound oxidation in environments in which organic compounds are present.« less

  6. Identification of Lactic Acid Bacteria in Fruit Pulp Processing Byproducts and Potential Probiotic Properties of Selected Lactobacillus Strains

    PubMed Central

    Garcia, Estefânia F.; Luciano, Winnie A.; Xavier, Danilo E.; da Costa, Whyara C. A.; de Sousa Oliveira, Kleber; Franco, Octávio L.; de Morais Júnior, Marcos A.; Lucena, Brígida T. L.; Picão, Renata C.; Magnani, Marciane; Saarela, Maria; de Souza, Evandro L.

    2016-01-01

    This study aimed to identify lactic acid bacteria (LAB) in byproducts of fruit (Malpighia glabra L., Mangifera indica L., Annona muricata L., and Fragaria vesca L.) pulp processing. Fifty strains of LAB were identified using matrix-assisted laser desorption/ionization–time of flight mass spectrometry (MALDI-TOF MS) and 16S rRNA gene sequence (16S rRNA) analysis. Species belonging to Lactobacillus genus were the predominant LAB in all fruit pulp processing byproducts. The average congruency between the MALDI-TOF MS and 16S rRNA in LAB species identification reached 86%. Isolates of L. plantarum, L. brevis, L. pentosus, L. lactis and L. mesenteroides were identified with 100% congruency. MALDI-TOF MS and 16S rRNA analysis presented 86 and 100% efficiency of LAB species identification, respectively. Further, five selected Lactobacillus strains (L. brevis 59, L. pentosus 129, L. paracasei 108, L. plantarum 49, and L. fermentum 111) were evaluated for desirable probiotic-related properties and growth behavior on two different cultivation media. The exposure to pH 2.0 sharply decreased the counts of the different Lactobacillus strains after a 1 or 2 h incubation, while varied decreases were noted after 3 h of exposure to pH 3.0. Overall, the exposure to pH 5.0 and to bile salts (0.15, 0.30, and 1.00%) did not decrease the counts of the Lactobacillus strains. All tested Lactobacillus strains presented inhibitory activity against Staphylococcus aureus, Salmonella Typhimurium, Salmonella Enteritidis, Listeria monocytogenes and Escherichia coli, and presented variable susceptibility to different antibiotics. The selected Lactobacillus strains presented satisfactory and reproducible growth behavior. In conclusion, MALDI-TOF MS and 16S rRNA analysis revealed high efficiency and congruency for LAB species identification, and the selected Lactobacillus strains may be candidates for further investigation of novel probiotic strains. PMID:27625647

  7. Rapid identification of 11 human intestinal Lactobacillus species by multiplex PCR assays using group- and species-specific primers derived from the 16S-23S rRNA intergenic spacer region and its flanking 23S rRNA.

    PubMed

    Song, Y; Kato, N; Liu, C; Matsumiya, Y; Kato, H; Watanabe, K

    2000-06-15

    Rapid and reliable two-step multiplex polymerase chain reaction (PCR) assays were established to identify human intestinal lactobacilli; a multiplex PCR was used for grouping of lactobacilli with a mixture of group-specific primers followed by four multiplex PCR assays with four sorts of species-specific primer mixtures for identification at the species level. Primers used were designed from nucleotide sequences of the 16S-23S rRNA intergenic spacer region and its flanking 23S rRNA gene of members of the genus Lactobacillus which are commonly isolated from human stool specimens: Lactobacillus acidophilus, Lactobacillus crispatus, Lactobacillus delbrueckii (ssp. bulgaricus and ssp. lactis), Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus jensenii, Lactobacillus paracasei (ssp. paracasei and ssp. tolerans), Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus rhamnosus and Lactobacillus salivarius (ssp. salicinius and ssp. salivarius). The established two-step multiplex PCR assays were applied to the identification of 84 Lactobacillus strains isolated from human stool specimens and the PCR results were consistent with the results from the DNA-DNA hybridization assay. These results suggest that the multiplex PCR system established in this study is a simple, rapid and reliable method for the identification of common Lactobacillus isolates from human stool samples.

  8. Upregulation of P-glycoprotein by probiotics in intestinal epithelial cells and in the dextran sulfate sodium model of colitis in mice

    PubMed Central

    Goyal, Sonia; Raheja, Geetu; Singh, Varsha; Akhtar, Maria; Nazir, Talat M.; Alrefai, Waddah A.; Gill, Ravinder K.; Dudeja, Pradeep K.

    2011-01-01

    P-glycoprotein (P-gp) mediates efflux of xenobiotics and bacterial toxins from the intestinal mucosa into the lumen. Dysregulation of P-gp has been implicated in inflammatory bowel disease. Certain probiotics have been shown to be effective in treating inflammatory bowel disease. However, direct effects of probiotics on P-gp are not known. Current studies examined the effects of Lactobacilli on P-gp function and expression in intestinal epithelial cells. Caco-2 monolayers and a mouse model of dextran sulfate sodium-induced colitis were utilized. P-gp activity was measured as verapamil-sensitive [3H]digoxin transepithelial flux. Multidrug resistant 1 (MDR1)/P-gp expression was measured by real-time quantitative PCR and immunoblotting. Culture supernatant (CS; 1:10 or 1:50, 24 h) of Lactobacillus acidophilus or Lactobacillus rhamnosus treatment of differentiated Caco-2 monolayers (21 days postplating) increased (∼3-fold) MDR1/P-gp mRNA and protein levels. L. acidophilus or L. rhamnosus CS stimulated P-gp activity (∼2-fold, P < 0.05) via phosphoinositide 3-kinase and ERK1/2 MAPK pathways. In mice, L. acidophilus or L. rhamnosus treatment (3 × 109 colony-forming units) increased mdr1a/P-gp mRNA and protein expression in the ileum and colon (2- to 3-fold). In the dextran sulfate sodium (DSS)-induced colitis model (3% DSS in drinking water for 7 days), the degree of colitis as judged by histological damage and myeloperoxidase activity was reduced by L. acidophilus. L. acidophilus treatment to DSS-treated mice blocked the reduced expression of mdr1a/P-gp mRNA and protein in the distal colon. These findings suggest that Lactobacilli or their soluble factors stimulate P-gp expression and function under normal and inflammatory conditions. These data provide insights into a novel mechanism involving P-gp upregulation in beneficial effects of probiotics in intestinal inflammatory disorders. PMID:21350189

  9. Comparative genomics of Lactobacillus

    PubMed Central

    Kant, Ravi; Blom, Jochen; Palva, Airi; Siezen, Roland J.; de Vos, Willem M.

    2011-01-01

    Summary The genus Lactobacillus includes a diverse group of bacteria consisting of many species that are associated with fermentations of plants, meat or milk. In addition, various lactobacilli are natural inhabitants of the intestinal tract of humans and other animals. Finally, several Lactobacillus strains are marketed as probiotics as their consumption can confer a health benefit to host. Presently, 154 Lactobacillus species are known and a growing fraction of these are subject to draft genome sequencing. However, complete genome sequences are needed to provide a platform for detailed genomic comparisons. Therefore, we selected a total of 20 genomes of various Lactobacillus strains for which complete genomic sequences have been reported. These genomes had sizes varying from 1.8 to 3.3 Mb and other characteristic features, such as G+C content that ranged from 33% to 51%. The Lactobacillus pan genome was found to consist of approximately 14 000 protein‐encoding genes while all 20 genomes shared a total of 383 sets of orthologous genes that defined the Lactobacillus core genome (LCG). Based on advanced phylogeny of the proteins encoded by this LCG, we grouped the 20 strains into three main groups and defined core group genes present in all genomes of a single group, signature group genes shared in all genomes of one group but absent in all other Lactobacillus genomes, and Group‐specific ORFans present in core group genes of one group and absent in all other complete genomes. The latter are of specific value in defining the different groups of genomes. The study provides a platform for present individual comparisons as well as future analysis of new Lactobacillus genomes. PMID:21375712

  10. Synthesis, Structure, Bonding, and Reactivity of Metal Complexes Comprising Diborane(4) and Diborene(2): [{Cp*Mo(CO)2 }2 {μ-η22 -B2 H4 }] and [{Cp*M(CO)2 }2 B2 H2 M(CO)4 ], M=Mo,W.

    PubMed

    Mondal, Bijan; Bag, Ranjit; Ghorai, Sagar; Bakthavachalam, K; Jemmis, Eluvathingal D; Ghosh, Sundargopal

    2018-07-02

    The reaction of [(Cp*Mo) 2 (μ-Cl) 2 B 2 H 6 ] (1) with CO at room temperature led to the formation of the highly fluxional species [{Cp*Mo(CO) 2 } 2 {μ-η 22 -B 2 H 4 }] (2). Compound 2, to the best of our knowledge, is the first example of a bimetallic diborane(4) conforming to a singly bridged C s structure. Theoretical studies show that 2 mimics the Cotton dimolybdenum-alkyne complex [{CpMo(CO) 2 } 2 C 2 H 2 ]. In an attempt to replace two hydrogen atoms of diborane(4) in 2 with a 2e [W(CO) 4 ] fragment, [{Cp*Mo(CO) 2 } 2 B 2 H 2 W(CO) 4 ] (3) was isolated upon treatment with [W(CO) 5 ⋅thf]. Compound 3 shows the intriguing presence of [B 2 H 2 ] with a short B-B length of 1.624(4) Å. We isolated the tungsten analogues of 3, [{Cp*W(CO) 2 } 2 B 2 H 2 W(CO) 4 ] (4) and [{Cp*W(CO) 2 } 2 B 2 H 2 Mo(CO) 4 ] (5), which provided direct proof of the existence of the tungsten analogue of 2. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Growth and Survival of Some Probiotic Strains in Simulated Ice Cream Conditions

    NASA Astrophysics Data System (ADS)

    Homayouni, A.; Ehsani, M. R.; Azizi, A.; Razavi, S. H.; Yarmand, M. S.

    A Completely Randomized Design (CRD) experiment was applied in triplicates to evaluate the survival of four probiotic strains in simulated ice cream conditions. The growth and survival rate of these probiotic strains (Lactobacillus acidophilus, Lactobacillus casei, Bifidobacterium bifidum and Bifidobacterium longum) in varying amount of sucrose (10, 15, 20 and 25%), oxygen scavenging components (0.05% L-cysteine and 0.05% L-ascorbate) and temperatures (4 and -20°C) during different periods of time (1, 2 and 3 months) were evaluated in MRS-broth medium. Optical density at 580 nm was used to measure growth. Lactobacilli strains proved to be highly resistant in comparison with Biffidobacteria strains. The viable cell number of Lactobacillus casei in different sucrose concentrations, different oxidoreduction potentials and refrigeration temperature was 1x1010, 2x108 and 5x107 cfu mL-1, respectively. Growth and survival rate of Lactobacillus casei showed to be the highest.

  12. Complete Genome Sequences of Lactobacillus johnsonii Strain N6.2 and Lactobacillus reuteri Strain TD1.

    PubMed

    Leonard, Michael T; Valladares, Ricardo B; Ardissone, Alexandria; Gonzalez, Claudio F; Lorca, Graciela L; Triplett, Eric W

    2014-05-08

    We report here the complete genome sequences of Lactobacillus johnsonii strain N6.2, a homofermentative lactic acid intestinal bacterium, and Lactobacillus reuteri strain TD1, a heterofermentative lactic acid intestinal bacterium, both isolated from a type 1 diabetes-resistant rat model.

  13. Lactobacillus fermentum ATCC 23271 Displays In vitro Inhibitory Activities against Candida spp.

    PubMed Central

    do Carmo, Monique S.; Noronha, Francisca M. F.; Arruda, Mariana O.; Costa, Ênnio P. da Silva; Bomfim, Maria R. Q.; Monteiro, Andrea S.; Ferro, Thiago A. F.; Fernandes, Elizabeth S.; Girón, Jorge A.; Monteiro-Neto, Valério

    2016-01-01

    Lactobacilli are involved in the microbial homeostasis in the female genital tract. Due to the high prevalence of many bacterial diseases of the female genital tract and the resistance of microorganisms to various antimicrobial agents, alternative means to control these infections are necessary. Thus, this study aimed to evaluate the probiotic properties of well-characterized Lactobacillus species, including L. acidophilus (ATCC 4356), L. brevis (ATCC 367), L. delbrueckii ssp. delbrueckii (ATCC 9645), L. fermentum (ATCC 23271), L. paracasei (ATCC 335), L. plantarum (ATCC 8014), and L. rhamnosus (ATCC 9595), against Candida albicans (ATCC 18804), Neisseria gonorrhoeae (ATCC 9826), and Streptococcus agalactiae (ATCC 13813). The probiotic potential was investigated by using the following criteria: (i) adhesion to host epithelial cells and mucus, (ii) biofilm formation, (iii) co-aggregation with bacterial pathogens, (iv) inhibition of pathogen adhesion to mucus and HeLa cells, and (v) antimicrobial activity. Tested lactobacilli adhered to mucin, co-aggregated with all genital microorganisms, and displayed antimicrobial activity. With the exception of L. acidophilus and L. paracasei, they adhered to HeLa cells. However, only L. fermentum produced a moderate biofilm and a higher level of co-aggregation and mucin binding. The displacement assay demonstrated that all Lactobacillus strains inhibit C. albicans binding to mucin (p < 0.001), likely due to the production of substances with antimicrobial activity. Clinical isolates belonging to the most common Candida species associated to vaginal candidiasis were inhibited by L. fermentum. Collectively, our data suggest that L. fermentum ATCC 23271 is a potential probiotic candidate, particularly to complement candidiasis treatment, since presented with the best probiotic profile in comparison with the other tested lactobacilli strains. PMID:27833605

  14. A mixture of Lactobacillus casei, Lactobacillus lactis, and Paenibacillus polymyxa reduces Escherichia coli O157:H7 in finishing feedlot cattle.

    PubMed

    Stanford, Kim; Bach, Susan; Baah, John; McAllister, Tim

    2014-05-01

    A direct-fed microbial (DFM) containing Paenibacillus polymyxa, Lactobacillus casei, and Lactobacillus lactis was fed to cattle (n = 120) to determine impacts on shedding and survival of Escherichia coli O157:H7 in feces. Cattle were individually penned and fed diets containing 0 (control), 4 × 10(7) CFU (DFM-4), 8 × 10(7) CFU (DFM-8), or 1.2 × 10(8) CFU (DFM-12) lactobacilli per kg of dietary dry matter over 84-day fall-winter growing and 140-day spring-summer finishing periods. Fecal grab samples were collected from cattle at 28-day intervals, E. coli O157:H7 was detected by immunomagnetic separation, and isolates were compared by pulsed-field gel electrophoresis. During the growing period, feces negative for E. coli O157 from each dietary treatment were inoculated with 10(5) CFU/g nalidixic acid-resistant E. coli O157:H7 and were incubated at 4 and 22(u) C for 11 weeks. Fecal pH and fecal dry matter were measured on days 0, 1, 3, and 7 and weekly thereafter, with E. coli O157:H7 enumerated through dilution plating. Treatment with DFMs did not affect survival of E. coli O157:H7 in feces or fecal pH (P > 0.05). Only one steer was positive for E. coli O157:H7 during the growing period, but during the finishing period, DFM-8 and DFM-12 reduced the prevalence of E. coli O157:H7 in feces (P < 0.05). Feeding DFMs also reduced the frequency of individual steers shedding E. coli O157:H7 during finishing (P < 0.05), with control steers shedding E. coli O157:H7 up to four times, whereas DFM-12 steers shed E. coli O157:H7 a maximum of twice. Treatment with DFMs influenced pulsed-field gel electrophoresis profiles; steers that were fed DFM-8 and DFM-12 shed more diverse subtypes of E. coli O157:H7 than did control or DFM-4 steers. Because a companion study found linear improvement in performance with increasing dosage of DFMs in the first 28 days of the growing period, targeted use of DFM-12 during this time and for the final 1 or 2 weeks prior to slaughter may optimize

  15. 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

    2015-01-01

    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.

  16. Effect of phenolic compounds on the growth of selected probiotic and pathogenic bacteria.

    PubMed

    Pacheco-Ordaz, R; Wall-Medrano, A; Goñi, M G; Ramos-Clamont-Montfort, G; Ayala-Zavala, J F; González-Aguilar, G A

    2018-01-01

    Fruit extracts from different tissues (pulp, seed and peel) have shown antimicrobial and prebiotic activities related to their phenolic profile, although structure-specific evaluations have not been reported yet. The effect of five phenolic compounds (catechin and gallic, vanillic, ferulic and protocatechuic acids) identified in different fruits, particularly in mango, was evaluated on the growth of two probiotic (Lactobacillus rhamnosusGG ATCC 53103 and Lactobacillus acidophilusNRRLB 4495) and two pathogenic (Escherichia coli 0157:H7 ATCC 43890 and Salmonella enterica serovar Typhimurium ATCC 14028) bacteria. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of phenolic acids ranged from 15-20 mmol l -1 and 20-30 mmol l -1 against E. coli and S. Typhimurium, respectively. For catechin, the MIC and MBC were 35 mmol l -1 and >35 mmol l -1 against E. coli and S. Typhimurium, respectively. The presence of catechin and gallic, protocatechuic and vanillic acids in MRS broth without dextrose allowed the growth of lactobacilli. Catechin combined with protocatechuic or vanillic acid mildly allowed the growth of both probiotics. In conclusion, phenolic compounds can selectively inhibit the growth of pathogenic bacteria without affecting the viability of probiotics. This study provides relevant information about the effects of phenolic compounds commonly present in fruit and vegetables on the growth of probiotic and pathogenic bacteria. The compounds selectively allowed the growth of probiotic lactobacilli (Lactobacillus rhamnosus GG and Lactobacillus acidophilus) and inhibited pathogenic bacteria (Escherichia coli and Salmonella Typhimurium) at the same concentration (20 mmol l -1 ). These findings can contribute to the formulation of nutraceutical products, such as synbiotics, that can restore or maintain an optimal composition of human microbiota, potentially improving the overall health of the consumer. © 2017 The

  17. Assessment and comparison of probiotic potential of four Lactobacillus species isolated from feces samples of Iranian infants.

    PubMed

    Halimi, Shahnaz; Mirsalehian, Akbar

    2016-02-01

    The probiotic potential of Lactobacillus species isolated from infant feces was investigated. For this study, the antibiotic susceptibility, tolerance in gut-related conditions, antimicrobial activity, and ability to adhere to a human colorectal adenocarcinoma cell line (Caco-2 cells) of four common Lactobacillus species (Lactobacillus paracasei [n = 15], Lactobacillus rhamnosus [n = 45], Lactobacillus gasseri [n = 20] and Lactobacillus fermentum [n = 18]) were assessed. Most isolates that which were sensitive to imipenem, ampicillin, gentamycin, erythromycin and tetracycline were selected for other tests. L. gasseri isolates had the greatest sensitivity to gastric and intestinal fluids (<10% viability). L. fermentum (FH5, FH13 and FH18) had the highest adhesion to Caco-2 cells. The lowest antibacterial activity against pathogenic bacteria was shown by L. gasseri strains in spot tests. Furthermore, non-adjusted cell-free culture supernatants with low pH had greater antimicrobial activity, which was related to organic acid. The results showed that some isolates of L. rhamnosus and L. fermentum are suitable for use as a probiotic. © 2015 The Societies and John Wiley & Sons Australia, Ltd.

  18. 77 FR 59670 - Electronic Filing of H-2A and H-2B Labor Certification Applications Through the iCERT Visa Portal...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-09-28

    ... DEPARTMENT OF LABOR Employment and Training Administration Electronic Filing of H-2A and H-2B... certification applications under the H-2A and H-2B visa programs through the Department of Labor's (Department... will be able to submit H-2B applications electronically beginning on October 15, 2012, and H-2A...

  19. Identification of Human Intestinal Bacteria that Promote or Inhibit Inflammation

    DTIC Science & Technology

    2012-11-01

    Lactobacillus "acidophilus" Lactobacillus " plantarum " ProvidenCa"sp." Enterobacter"aerogenes" Lactobacillus "paraalimentarius" Salmonella"typhi...8217 indica3ve’of’the’poten3al’of’these’microbes’to’induce’intes3nal’inflamma3on.’ 0" 5" 10" 15" 20" 25" 30" 35" 40" 45" 50" no"bacteria" Lactobacillus "brevis" Escherichea"coli"(DH5a)" Klebsiella"pneumonia" Salmonella...only

  20. Lactobacillus delbrueckii ssp. bulgaricus B-30892 can inhibit cytotoxic effects and adhesion of pathogenic Clostridium difficile to Caco-2 cells

    PubMed Central

    Banerjee, Pratik; Merkel, Glenn J; Bhunia, Arun K

    2009-01-01

    Background Probiotic microorganisms are receiving increasing interest for use in the prevention, treatment, or dietary management of certain diseases, including antibiotic-associated diarrhea (AAD). Clostridium difficile is the most common cause of AAD and the resulting C. difficile – mediated infection (CDI), is potentially deadly. C. difficile associated diarrhea (CDAD) is manifested by severe inflammation and colitis, mostly due to the release of two exotoxins by C. difficile causing destruction of epithelial cells in the intestine. The aim of this study was to determine the effect of probiotic bacteria Lactobacillus delbrueckii ssp. bulgaricus B-30892 (LDB B-30892) on C. difficile-mediated cytotoxicity using Caco-2 cells as a model. Methods Experiments were carried out to test if the cytotoxicity induced by C. difficile-conditioned-medium on Caco-2 cells can be altered by cell-free supernatant (CFS) from LDB B-30892 in different dilutions (1:2 to 1:2048). In a similar experimental setup, comparative evaluations of other probiotic strains were made by contrasting the results from these strains with the results from LDB B-30892, specifically the ability to affect C. difficile induced cytotoxicity on Caco-2 monolayers. Adhesion assays followed by quantitative analysis by Giemsa staining were conducted to test if the CFSs from LDB B-30892 and other probiotic test strains have the capability to alter the adhesion of C. difficile to the Caco-2 monolayer. Experiments were also performed to evaluate if LDB B-30892 or its released components have any bactericidal effect on C. difficile. Results and discussion Co-culturing of LDB B-30892 with C. difficile inhibited the C. difficile-mediated cytotoxicity on Caco-2 cells. When CFS from LDB B-30892-C. difficile co-culture was administered (up to a dilution of 1:16) on Caco-2 monolayer, there were no signs of cytotoxicity. When CFS from separately grown LDB B-30892 was mixed with the cell-free toxin preparation (CFT) of

  1. Structural basis for histone H2B deubiquitination by the SAGA DUB module

    DOE PAGES

    Morgan, Michael T.; Haj-Yahya, Mahmood; Ringel, Alison E.; ...

    2016-02-12

    Monoubiquitinated histone H2B plays multiple roles in transcription activation. H2B is deubiquitinated by the Spt-Ada-Gcn5 acetyltransferase (SAGA) coactivator, which contains a four-protein subcomplex known as the deubiquitinating (DUB) module. In this paper, the crystal structure of the Ubp8/Sgf11/Sus1/Sgf73 DUB module bound to a ubiquitinated nucleosome reveals that the DUB module primarily contacts H2A/H2B, with an arginine cluster on the Sgf11 zinc finger domain docking on the conserved H2A/H2B acidic patch. The Ubp8 catalytic domain mediates additional contacts with H2B, as well as with the conjugated ubiquitin. Finally, we find that the DUB module deubiquitinates H2B both in the context ofmore » the nucleosome and in H2A/H2B dimers complexed with the histone chaperone, FACT, suggesting that SAGA could target H2B at multiple stages of nucleosome disassembly and reassembly during transcription.« less

  2. Role of plnB gene in the regulation of bacteriocin production in Lactobacillus paraplantarum L-XM1.

    PubMed

    Zhang, Xiangmei; Shang, Nan; Zhang, Xu; Gui, Meng; Li, Pinglan

    2013-06-12

    Homologues of plnB gene have been shown to participate in regulation of bacteriocin production through quorum sensing system in other organisms, to investigate the possible role of plnB gene in Lactobacillus paraplantarum L-XM1, we cloned and insertionally inactivated the plnB gene. The plnB knockout mutant ΔplnB21 showed loss of bacteriocin production, its Bac⁺ phenotype could not be restored even after the addition of PlnA. Furthermore, reverse transcription-PCR analysis from total RNA preparations showed that the bacteriocin structural genes of the plnEF and plnJK were not transcribed in the plnB knockout mutant compared with the wild-type strain. It was therefore concluded that plnB is invovled in a quorum sensing based bacteriocin production. This is the first demonstration of a role for plnB by gene knockout in L. paraplantarum. Copyright © 2012 Elsevier GmbH. All rights reserved.

  3. Dynamics of the formation and loss of boron atoms in a H2/B2H6 microwave plasma

    NASA Astrophysics Data System (ADS)

    Duluard, C. Y.; Aubert, X.; Sadeghi, N.; Gicquel, A.

    2016-09-01

    For further improvements in doped-diamond deposition technology, an understanding of the complex chemistry in H2/CH4/B2H6 plasmas is of general importance. In this context, a H2/B2H6 plasma ignited by microwave power in a near resonant cavity at high pressure (100-200 mbar) is studied to measure the B-atom density in the ground state. The discharge is ignited in the gas mixture (0-135 ppm B2H6 in H2) by a 2.45 GHz microwave generator, leading to the formation of a hemispheric plasma core, surrounded by a faint discharge halo filling the remaining reactor volume. Measurements with both laser induced fluorescence and resonant absoption with a boron hollow cathode lamp indicate that the B-atom density is higher in the halo than in the plasma core. When the absorption line-of-sight is positioned in the halo, the absorption is so strong that the upper detection limit is reached. To understand the mechanisms of creation and loss of boron atoms, time-resolved absorption measurements have been carried out in a pulsed plasma regime (10 Hz, duty cycle 50%). The study focuses on the influence of the total pressure, the partial pressure of B2H6, as well as the source power, on the growth and decay rates of boron atoms when the plasma is turned off.

  4. Cholesterol-Lowering Potentials of Lactic Acid Bacteria Based on Bile-Salt Hydrolase Activity and Effect of Potent Strains on Cholesterol Metabolism In Vitro and In Vivo

    PubMed Central

    Lin, Pei-Pei; Hsieh, You-Miin; Zhang, Zi-yi; Wu, Hui-Ching; Huang, Chun-Chih

    2014-01-01

    This study collected different probiotic isolates from animal and plant sources to evaluate the bile-salt hydrolase activity of probiotics in vitro. The deconjugation potential of bile acid was determined using high-performance liquid chromatography. HepG2 cells were cultured with probiotic strains with high BSH activity. The triglyceride (TG) and apolipoprotein B (apo B) secretion by HepG2 cells were evaluated. Our results show that the BSH activity and bile-acid deconjugation abilities of Pediococcus acidilactici NBHK002, Bifidobacterium adolescentis NBHK006, Lactobacillus rhamnosus NBHK007, and Lactobacillus acidophilus NBHK008 were higher than those of the other probiotic strains. The cholesterol concentration in cholesterol micelles was reduced within 24 h. NBHK007 reduced the TG secretion by 100% after 48 h of incubation. NBHK002, NBHK006, and NBHK007 could reduce apo B secretion by 33%, 38%, and 39%, respectively, after 24 h of incubation. The product PROBIO S-23 produced a greater decrease in the total concentration of cholesterol, low-density lipoprotein, TG, and thiobarbituric acid reactive substance in the serum or livers of hamsters with hypercholesterolemia compared with that of hamsters fed with a high-fat and high-cholesterol diet. These results show that the three probiotic strains of lactic acid bacteria are better candidates for reducing the risk of cardiovascular disease. PMID:25538960

  5. Topology-energy relationships and lowest energy configurations for pentagonal dodecahedral (H2O)20X clusters, X=empty, H2O, NH3, H3O+: The importance of O-topology

    NASA Astrophysics Data System (ADS)

    Anick, David J.

    2010-04-01

    For (H2O)20X water clusters consisting of X enclosed by the 512 dodecahedral cage, X=empty, H2O, NH3, and H3O+, databases are made consisting of 55-82 isomers optimized via B3LYP/6-311++G∗∗. Correlations are explored between ground state electronic energy (Ee) or electronic energy plus zero point energy (Ee+ZPE) and the clusters' topology, defined as the set of directed H-bonds. Linear regression is done to identify topological features that correlate with cluster energy. For each X, variables are found that account for 99% of the variance in Ee and predict it with a rms error under 0.2 kcal/mol. The method of analysis emphasizes the importance of an intermediate level of structure, the "O-topology," consisting of O-types and a list of O pairs that are bonded but omitting H-bond directions, as a device to organize the databases and reduce the number of structures one needs to consider. Relevant variables include three parameters, which count the number of H-bonds having particular donor and acceptor types; |M|2, where M is the cluster's vector dipole moment; and the projection of M onto the symmetry axis of X. Scatter diagrams for Ee or Ee+ZPE versus |M| show that clusters fall naturally into "families" defined by the values of certain discrete parameters, the "major parameters," for each X. Combining "family" analysis and O-topologies, a small group of clusters is identified for each X that are candidates to be the global minimum, and the minimum is determined. For X=H3O+, one cluster with central hydronium lies just 2.08 kcal/mol above the lowest isomer with surface hydronium. Implications of the methodology for dodecahedral (H2O)20(NH4+) and (H2O)20(NH4+)(OH-) are discussed, and new lower energy isomers are found. For MP2/TZVP, the lowest-energy (H2O)20(NH4+) isomer features a trifurcated H-bond. The results suggest a much more efficient and comprehensive way of seeking low-energy water cluster geometries that may have wide applicability.

  6. Topology-energy relationships and lowest energy configurations for pentagonal dodecahedral (H2O)20X clusters, X = empty, H2O, NH3, H3O+: the importance of O-topology.

    PubMed

    Anick, David J

    2010-04-28

    For (H(2)O)(20)X water clusters consisting of X enclosed by the 5(12) dodecahedral cage, X = empty, H(2)O, NH(3), and H(3)O(+), databases are made consisting of 55-82 isomers optimized via B3LYP/6-311++G(**). Correlations are explored between ground state electronic energy (Ee) or electronic energy plus zero point energy (Ee+ZPE) and the clusters' topology, defined as the set of directed H-bonds. Linear regression is done to identify topological features that correlate with cluster energy. For each X, variables are found that account for 99% of the variance in Ee and predict it with a rms error under 0.2 kcal/mol. The method of analysis emphasizes the importance of an intermediate level of structure, the "O-topology," consisting of O-types and a list of O pairs that are bonded but omitting H-bond directions, as a device to organize the databases and reduce the number of structures one needs to consider. Relevant variables include three parameters, which count the number of H-bonds having particular donor and acceptor types; absolute value(M)(2), where M is the cluster's vector dipole moment; and the projection of M onto the symmetry axis of X. Scatter diagrams for Ee or Ee+ZPE versus absolute value(M) show that clusters fall naturally into "families" defined by the values of certain discrete parameters, the "major parameters," for each X. Combining "family" analysis and O-topologies, a small group of clusters is identified for each X that are candidates to be the global minimum, and the minimum is determined. For X = H(3)O(+), one cluster with central hydronium lies just 2.08 kcal/mol above the lowest isomer with surface hydronium. Implications of the methodology for dodecahedral (H(2)O)(20)(NH(4)(+)) and (H(2)O)(20)(NH(4)(+))(OH(-)) are discussed, and new lower energy isomers are found. For MP2/TZVP, the lowest-energy (H(2)O)(20)(NH(4)(+)) isomer features a trifurcated H-bond. The results suggest a much more efficient and comprehensive way of seeking low

  7. Proteome analysis of Lactobacillus helveticus H9 during growth in skim milk.

    PubMed

    Chen, Y F; Zhao, W J; Wu, R N; Sun, Z H; Zhang, W Y; Wang, J C; Bilige, M; Zhang, H P

    2014-12-01

    Lactobacillus helveticus H9 was isolated from traditionally fermented yak milk in Tibet (China) with the ability to produce the antihypertensive peptides Val-Pro-Pro (VPP) and Ile-Pro-Pro (IPP) during milk fermentation. To understand the changes in the protein expression of L. helveticus H9, proteome analysis was performed at 3 different growth stages, lag phase (pH 6.1), log phase (pH 5.1), and stationary phase (pH 4.5) using 2-dimensional electrophoresis (2-DE). Further analysis showed that 257 differential protein spots were found and 214 protein spots were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS). The cellular functions of the differentially expressed proteins were complex. Interestingly, the proteolytic system-related proteins aminopeptidase N (PepN), aminopeptidase E (PepE), endopeptidase O2 (PepO2), and oligopeptide transport system permease protein (OppC) were observed only on the maps of pH 5.1 and pH 4.5, which was consistent with the presence of angiotensin I-converting enzyme (ACE)-inhibitory peptides VPP and IPP during these 2 growth stages (log phase and stationary phase). These results, combined with a previous study of gene expression of the proteolytic system, led us to conclude that the Opp transport system, pepE, and pepO2 are likely related to the production of ACE-inhibitory peptides. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  8. Mutational Studies Uncover Non-Native Structure in the Dimeric Kinetic Intermediate of the H2A-H2B Heterodimer

    PubMed Central

    Stump, Matthew R.; Gloss, Lisa M.

    2010-01-01

    The folding pathway of the histone H2A-H2B heterodimer minimally includes an on-pathway, dimeric, burst-phase intermediate, I2. The partially folded H2A and H2B monomers populated at equilibrium were characterized as potential monomeric kinetic intermediates. Folding kinetics were compared for initiation from isolated, folded monomers and the heterodimer unfolded in 4 M urea. The observed rates were virtually identical above 0.4 M urea, exhibiting a log-linear relationship on the final denaturant concentration. Below ~0.4 M urea (concentrations inaccessible from the 4 M urea unfolded state), a roll-over in the rates was observed; this suggests that a component of the I2 ensemble contains non-native structure that rearranges/isomerizes to a more native-like species. The contribution of helix propensity to the stability of the I2 ensemble was assessed with a set of H2A-H2B mutants containing Ala and Gly replacements at nine sites, focusing mainly on the long, central α2 helix. Equilibrium and kinetic folding/unfolding data were collected to determine the effects of the mutations on the stability of I2 and the transition state between I2 and N2. This limited mutational study indicated that residues in the α2 helices of H2A and H2B, as well as α1 of H2B and both the C-terminus of α3 and the short αC helix of H2A contribute to the stability of the I2 burst phase species. Interestingly, at least eight of the nine targeted residues stabilize I2 by interactions that are non-native to some extent. Given that destabilizing I2 and these non-native interactions does not accelerate folding, it is concluded that the native and non-native structure present in the I2 ensemble enables efficient folding of H2A-H2B. PMID:20600120

  9. Effects of inulin and oligofructose on the rheological characteristics and probiotic culture survival in low-fat probiotic ice cream.

    PubMed

    Akalin, A S; Erişir, D

    2008-05-01

    The effects of supplementation of oligofructose or inulin on the rheological characteristics and survival of Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 in low-fat ice cream stored at -18 degrees C for 90 d were studied. Addition of oligofructose or inulin to ice cream mix significantly increased apparent viscosity and overrun and developed the melting properties in ice cream during storage (P < 0.05). However, the highest increase in firmness, the lowest change in melting properties, and the longest 1st dripping time were obtained in probiotic ice cream containing inulin (P < 0.05). Some textural properties have also improved especially by the end of storage. Freezing process caused a significant decrease in the viability of Lactobacillus acidophilus La-5 and Bifidobacterium animalis Bb-12 (P < 0.05). Oligofructose significantly improved the viability of L. acidophilus La-5 and B. animalis Bb-12 in ice cream mix (P < 0.05). Although the viable numbers for both bacteria decreased throughout the storage, the minimum level of 10(6) CFU/g was maintained for B. animalis Bb-12 in only ice cream with oligofructose during storage.

  10. Different reaction of the core histones H2A and H2B to red laser irradiation

    NASA Astrophysics Data System (ADS)

    Brill, G. E.; Egorova, A. V.; Bugaeva, I. O.; Postnov, D. E.; Ushakova, O. V.

    2017-03-01

    Analysis of the influence of red laser irradiation on the processes of self-assembly of the core histones H2A and H2B was performed using a wedge dehydration method. Image-analysis of facies included their qualitative characteristics and calculation of quantitative parameters with subsequent statistical processing. It was established that linearly polarized red laser light (λ - 660 nm, 1 J/cm2) significantly modified the process of self-assembly of core histone H2B, whereas the structure of the facies of H2A histone changed to a lesser extent. Histones were used in the form of aqueous salt solutions. The effect of red light seems to result from the formation of singlet oxygen by direct laser excitation of molecular oxygen.

  11. Phase and Physicochemical Properties Diagrams of Quaternary System Li2B4O7 + Na2B4O7 + Mg2B6O11 + H2O

    NASA Astrophysics Data System (ADS)

    Wang, Shi-qiang; Du, Xue-min; Jing, Yan; Guo, Ya-fei; Deng, Tian-long

    2017-12-01

    The phase and physicochemical properties diagrams of the quaternary system (Li2B4O7 + Na2B4O7 + Mg2B6O11) at 288.15 K and 0.1 MPa were constructed using the solubilities, densities, and refractive indices measured. In the phase diagrams of the system there are one invariant point, three univariant isothermic dissolution curves, and three crystallization regions corresponding to Li2B4O7 · 3H2O, Na2B4O7 · 10H2O, and Mg2B6O11 · 15H2O, respectively. The solution density, refractive index of the quaternary system changes regularly with the increasing of Li2B4O7 concentration. The calculated values of density and refractive index using empirical equations of the quaternary system are in good agreement with the experimental values.

  12. ANTIFERROMAGNETIC SPIN RESONANCE OF COBR2.6H20,

    DTIC Science & Technology

    Electron spin resonance experiments were carried out on single crystals of CoBr2.6H20 at temperatures from 1.2 to 4.2K, using a microwave transmission spectrometer over a range of frequencies from 29 to 59 kMc .

  13. The extracellular phage-host interactions involved in the bacteriophage LL-H infection of Lactobacillus delbrueckii ssp. lactis ATCC 15808

    PubMed Central

    Munsch-Alatossava, Patricia; Alatossava, Tapani

    2013-01-01

    The complete genome sequence of Lactobacillus bacteriophage LL-H was determined in 1996. Accordingly, LL-H has been used as a model phage for the infection of dairy Lactobacillus, specifically for thermophilic Lactobacillus delbrueckii ssp. lactis host strains, such as ATCC 15808. One of the major goals of phage LL-H research consisted of the characterization of the first phage-host interactions at the level of phage adsorption and phage DNA injection steps to determine effective and practical methods to minimize the risks associated with the appearance and attack of phages in the manufacture of yogurt, and Swiss or Italian hard type cheeses, which typically use thermophilic lactic acid bacteria starter cultures containing L. delbrueckii strains among others. This mini review article summarizes the present data concerning (i) the special features, particle structure, and components of phage LL-H and (ii) the structure and properties of lipoteichoic acids (LTAs), which are the phage LL-H receptor components of L. delbrueckii ssp. lactis host strains. Moreover, a model of the first, extracellular, phage-host interactions for the infection of L. delbrueckii ssp. lactis ATCC 15808 by phage LL-H is presented and further discussed. PMID:24400001

  14. The extracellular phage-host interactions involved in the bacteriophage LL-H infection of Lactobacillus delbrueckii ssp. lactis ATCC 15808.

    PubMed

    Munsch-Alatossava, Patricia; Alatossava, Tapani

    2013-12-24

    The complete genome sequence of Lactobacillus bacteriophage LL-H was determined in 1996. Accordingly, LL-H has been used as a model phage for the infection of dairy Lactobacillus, specifically for thermophilic Lactobacillus delbrueckii ssp. lactis host strains, such as ATCC 15808. One of the major goals of phage LL-H research consisted of the characterization of the first phage-host interactions at the level of phage adsorption and phage DNA injection steps to determine effective and practical methods to minimize the risks associated with the appearance and attack of phages in the manufacture of yogurt, and Swiss or Italian hard type cheeses, which typically use thermophilic lactic acid bacteria starter cultures containing L. delbrueckii strains among others. This mini review article summarizes the present data concerning (i) the special features, particle structure, and components of phage LL-H and (ii) the structure and properties of lipoteichoic acids (LTAs), which are the phage LL-H receptor components of L. delbrueckii ssp. lactis host strains. Moreover, a model of the first, extracellular, phage-host interactions for the infection of L. delbrueckii ssp. lactis ATCC 15808 by phage LL-H is presented and further discussed.

  15. Transient MutS-Based Hypermutation System for Adaptive Evolution of Lactobacillus casei to Low pH.

    PubMed

    Overbeck, Tom J; Welker, Dennis L; Hughes, Joanne E; Steele, James L; Broadbent, Jeff R

    2017-10-15

    This study explored transient inactivation of the gene encoding the DNA mismatch repair enzyme MutS as a tool for adaptive evolution of Lactobacillus casei MutS deletion derivatives of L. casei 12A and ATCC 334 were constructed and subjected to a 100-day adaptive evolution process to increase lactic acid resistance at low pH. Wild-type parental strains were also subjected to this treatment. At the end of the process, the Δ mutS lesion was repaired in representative L. casei 12A and ATCC 334 Δ mutS mutant isolates. Growth studies in broth at pH 4.0 (titrated with lactic acid) showed that all four adapted strains grew more rapidly, to higher cell densities, and produced significantly more lactic acid than untreated wild-type cells. However, the adapted Δ mutS derivative mutants showed the greatest increases in growth and lactic acid production. Further characterization of the L. casei 12A-adapted Δ mutS derivative revealed that it had a significantly smaller cell volume, a rougher cell surface, and significantly better survival at pH 2.5 than parental L. casei 12A. Genome sequence analysis confirmed that transient mutS inactivation decreased DNA replication fidelity in both L. casei strains, and it identified genetic changes that might contribute to the lactic acid-resistant phenotypes of adapted cells. Targeted inactivation of three genes that had acquired nonsense mutations in the adapted L. casei 12A Δ mutS mutant derivative showed that NADH dehydrogenase ( ndh ), phosphate transport ATP-binding protein PstB ( pstB ), and two-component signal transduction system (TCS) quorum-sensing histidine protein kinase ( hpk ) genes act in combination to increase lactic acid resistance in L. casei 12A. IMPORTANCE Adaptive evolution has been applied to microorganisms to increase industrially desirable phenotypes, including acid resistance. We developed a method to increase the adaptability of Lactobacillus casei 12A and ATCC 334 through transient inactivation of the DNA

  16. Transient MutS-Based Hypermutation System for Adaptive Evolution of Lactobacillus casei to Low pH

    PubMed Central

    Overbeck, Tom J.; Welker, Dennis L.; Hughes, Joanne E.; Steele, James L.

    2017-01-01

    ABSTRACT This study explored transient inactivation of the gene encoding the DNA mismatch repair enzyme MutS as a tool for adaptive evolution of Lactobacillus casei. MutS deletion derivatives of L. casei 12A and ATCC 334 were constructed and subjected to a 100-day adaptive evolution process to increase lactic acid resistance at low pH. Wild-type parental strains were also subjected to this treatment. At the end of the process, the ΔmutS lesion was repaired in representative L. casei 12A and ATCC 334 ΔmutS mutant isolates. Growth studies in broth at pH 4.0 (titrated with lactic acid) showed that all four adapted strains grew more rapidly, to higher cell densities, and produced significantly more lactic acid than untreated wild-type cells. However, the adapted ΔmutS derivative mutants showed the greatest increases in growth and lactic acid production. Further characterization of the L. casei 12A-adapted ΔmutS derivative revealed that it had a significantly smaller cell volume, a rougher cell surface, and significantly better survival at pH 2.5 than parental L. casei 12A. Genome sequence analysis confirmed that transient mutS inactivation decreased DNA replication fidelity in both L. casei strains, and it identified genetic changes that might contribute to the lactic acid-resistant phenotypes of adapted cells. Targeted inactivation of three genes that had acquired nonsense mutations in the adapted L. casei 12A ΔmutS mutant derivative showed that NADH dehydrogenase (ndh), phosphate transport ATP-binding protein PstB (pstB), and two-component signal transduction system (TCS) quorum-sensing histidine protein kinase (hpk) genes act in combination to increase lactic acid resistance in L. casei 12A. IMPORTANCE Adaptive evolution has been applied to microorganisms to increase industrially desirable phenotypes, including acid resistance. We developed a method to increase the adaptability of Lactobacillus casei 12A and ATCC 334 through transient inactivation of the DNA

  17. [Correlation of Plasma Co-stimulatory Molecules B7-H2 and B7-H3 with Platelet Auto-antibodies in Patients with Immune Thrombocytopenic Purpura].

    PubMed

    Zuo, Bin; Zhao, Yun-Xiao; Yang, Jian-Feng; He, Yang

    2015-08-01

    To investigate whether the plasma level of platelet auto- antibodies in ITP patients is related to that of co-stimulatory molecules sB7-H2 and sB7-H3. A total of 61 ITP patients and 25 healthy controls from the First Affiliated Hospital of Soochow University from June 2012 to August 2013 were enrolled in this study. The expression levels of platelet auto-antibodies against 5 glycoproteins (GPIX, GP Ib, GP IIIa, GPIIb and P-selectin) in plasma were detected by flow cytometric immuno-beads array, and the expression of soluable co-stimulatory molecules sB7-H2 and sB7-H3 was measured by ELISA. The plasma levels of 5 auto-antibodies against platelet membrance glycoproteins significantly increased in ITP patiens (P < 0.01). Compared with healthy controls, sB7-H2 levels increased (P < 0.05), while the sB7-H3 level did not significantly change (r = 0.13, P > 0.05). However, the correlation analysis showed that sB7-H3 negatively correlated with platelet P-selectin auto-antibody (r = -0.46, P < 0.05), and sB7-H2 and sB7-H3 significantly reduced in ITP patients with positive P-selectin auto-antibody (P < 0.01). In ITP patients, platelet counts negatively correlated with sB7-H2 (r = -0.3907, P < 0.01), but did not correlate with sB7-H3. Soluble costimulatory molecule sB7-H2 elevates in ITP patients, and the level of sB7-H3 is associated with auto-antibodies against P-selectin, suggesting that costimulatory molecules B7-H2 and B7-H3 may be involved in the pathogenesis of immune regulation abnormality in ITP.

  18. Novel feruloyl esterase from Lactobacillus fermentum NRRL B-1932 and analysis of the recombinant enzyme produced in Escherichia coli.

    USDA-ARS?s Scientific Manuscript database

    Using agar plates containing ethyl ferulate as the sole carbon source, 33 Lactobacillus strains were screened for feruloyl esterase (FE) activity. Among a dozen species showing a clearing zone on the opaque plate containing ethyl ferulate, Lactobacillus fermentum NRRL B-1932 demonstrated the stronge...

  19. Crystallographic and mutational analyses of cystathionine β-synthase in the H2 S-synthetic gene cluster in Lactobacillus plantarum.

    PubMed

    Matoba, Yasuyuki; Yoshida, Tomoki; Izuhara-Kihara, Hisae; Noda, Masafumi; Sugiyama, Masanori

    2017-04-01

    Cystathionine β-synthase (CBS) catalyzes the formation of l-cystathionine from l-serine and l-homocysteine. The resulting l-cystathionine is decomposed into l-cysteine, ammonia, and α-ketobutylic acid by cystathionine γ-lyase (CGL). This reverse transsulfuration pathway, which is catalyzed by both enzymes, mainly occurs in eukaryotic cells. The eukaryotic CBS and CGL have recently been recognized as major physiological enzymes for the generation of hydrogen sulfide (H 2 S). In some bacteria, including the plant-derived lactic acid bacterium Lactobacillus plantarum, the CBS- and CGL-encoding genes form a cluster in their genomes. Inactivation of these enzymes has been reported to suppress H 2 S production in bacteria; interestingly, it has been shown that H 2 S suppression increases their susceptibility to various antibiotics. In the present study, we characterized the enzymatic properties of the L. plantarum CBS, whose amino acid sequence displays a similarity with those of O-acetyl-l-serine sulfhydrylase (OASS) that catalyzes the generation of l-cysteine from O-acetyl-l-serine (l-OAS) and H 2 S. The L. plantarum CBS shows l-OAS- and l-cysteine-dependent CBS activities together with OASS activity. Especially, it catalyzes the formation of H 2 S in the presence of l-cysteine and l-homocysteine, together with the formation of l-cystathionine. The high affinity toward l-cysteine as a first substrate and tendency to use l-homocysteine as a second substrate might be associated with its enzymatic ability to generate H 2 S. Crystallographic and mutational analyses of CBS indicate that the Ala70 and Glu223 residues at the substrate binding pocket are important for the H 2 S-generating activity. © 2017 The Protein Society.

  20. QM/MM Investigation of Substrate and Product Specificities of Suv4-20h2: How Does This Enzyme Generate Dimethylated H4K20 from Monomethylated Substrate?

    PubMed

    Qian, Ping; Guo, Haobo; Wang, Liang; Guo, Hong

    2017-06-13

    Protein lysine methyltransferases (PKMTs) catalyze the methylation of lysine residues on histone proteins in the regulation of chromatin structure and gene expression. In contrast to many other PKMTs for which unmodified lysine is the methylation target, the enzymes in the Suv4-20 family are able to generate dimethylated product (H4K20me2) based exclusively on the monomethylated H4K20 substrate (H4K20me1). The origin of such substrate/product specificity is still not clear. Here, molecular dynamics (MD) and free energy (potential of mean force) simulations are undertaken using quantum mechanical/molecular mechanical (QM/MM) potentials to understand the substrate/product specificities of Suv4-20h2, a member of the Suv4-20 family. The free energy barriers for mono-, di-, and trimethylation in Suv4-20h2 obtained from the simulations are found to be well correlated with the specificities observed experimentally with the allowed dimethylation based on the H4K20me1 substrate and prohibited monomethylation and trimethylation based on H4K20 and H4K20me2, respectively. It is demonstrated that the reason for the relatively efficient dimethylation is an effective transition state (TS) stabilization through strengthening the CH···O interactions as well as the presence of a cation-π interaction at the transition state. The simulations also show that the failures of Suv4-20h2 to catalyze monomethylation and trimethylation are due, respectively, to a less effective TS stabilization and inability of the reactant complex containing H4K20me2 to adopt a reactive (near attack) configuration for methyl transfer. The results suggest that care must be exercised in the prediction of the substrate specificity based only on the existence of near attack configurations in substrate complexes.

  1. Dehydrogenation reactions of cyclic C(2)B(2)N(2)H(12) and C(4)BNH(12) isomers.

    PubMed

    Matus, Myrna H; Liu, Shih-Yuan; Dixon, David A

    2010-02-25

    The energetics for different dehydrogenation pathways of C(2)B(2)N(2)H(12) and C(4)BNH(12) cycles were calculated at the B3LYP/DGDZVP2 and G3(MP2) levels with additional calculations at the CCSD(T)/complete basis set level. The heats of formation of the different isomers were calculated from the G3(MP2) relative energies and the heats of formation of the most stable isomers of c-C(2)B(2)N(2)H(6), c-C(2)B(2)N(2)H(12), and c-C(4)BNH(12) at the CCSD(T)/CBS including additional corrections together with the previously reported value for c-C(4)BNH(6). Different isomers were analyzed for c-C(2)B(2)N(2)H(x) and c-C(4)BNH(x) (x = 6 and 12), and the most stable cyclic structures were those with C-C-B-N-B-N and C-C-C-C-B-N sequences, respectively. The energetics for the stepwise loss of three H(2) were predicted, and the most feasible thermodynamic pathways were found. Dehydrogenation of the lowest energy c-C(2)B(2)N(2)H(12) isomer (6-H(12)) is almost thermoneutral with DeltaH(3dehydro) = 3.4 kcal/mol at the CCSD(T)/CBS level and -0.6 kcal/mol at the G3(MP2) level at 298 K. Dehydrogenation of the lowest energy c-C(4)BNH(12) isomer (7-H(12)) is endothermic with DeltaH(3dehydro) = 27.9 kcal/mol at the CCSD(T)/CBS level and 23.5 kcal/mol at the G3(MP2) level at 298 K. Dehydrogenation across the B-N bond is more favorable as opposed to dehydrogenation across the B-C, N-C, and C-C bonds. Resonance stabilization energies in relation to that of benzene are reported as are NICS NMR chemical shifts for correlating with the potential aromatic character of the rings.

  2. Probiotic properties of native Lactobacillus spp. strains for dairy calves.

    PubMed

    Fernández, S; Fraga, M; Silveyra, E; Trombert, A N; Rabaza, A; Pla, M; Zunino, P

    2018-04-10

    The use of native microorganisms with probiotic capacity is an alternative tool for the treatment and prevention of several diseases that affect animals, such as neonatal calf diarrhoea. The selection of probiotic strains within a collection is based on different in vitro and in vivo assays, which predict their potential. The aim of this study was to characterise a group of native Lactobacillus spp. strains isolated from faeces of healthy calves using an in vitro approach and to assess their ability to colonise the gastrointestinal tract (GIT) of calves. Native Lactobacillus spp. strains were evaluated on their capacity to survive low pH conditions and bile salts presence, biofilm formation and adhesion to both mucus and Caco-2 cells. Based on the in vitro characterisation, four strains (Lactobacillus johnsonii TP1.1, Lactobacillus reuteri TP1.3B, L. johnsonii TP1.6 and Lactobacillus amylovorus TP8.7) were selected to evaluate their capacity to colonise and persist in the GIT of calves. The assessment of enteric persistence involved an in vivo assay with oral administration of probiotics and quantification in faeces of the administered bacterial species with real-time quantitative PCR (qPCR). The study was conducted using 15 calves (1-month-old) which were divided into five groups of three animals, four of which were treated with four different selected strains and one was the control group. Strains TP1.3B and TP1.6 managed to persist in treated animals until ten days after the end of the administration period, indicating that they could be promising candidates for the design of probiotics for calves.

  3. Lactobacillus ixorae sp. nov., isolated from a flower (West-Indian jasmine).

    PubMed

    Techo, Sujitra; Miyashita, Mika; Shibata, Chiyo; Tanaka, Naoto; Wisetkhan, Preeyarach; Visessanguan, Wonnop; Tanasupawat, Somboon

    2016-12-01

    A Gram-stain-positive, lactic acid bacterium, strain Ru20-1T, was isolated from a flower (West-Indian jasmine) collected from Kalasin province, Thailand. A polyphasic approach was used to determine the taxonomic position of this strain. Studies of morphological and biochemical characteristics revealed that strain Ru20-1T belonged to the genus Lactobacillus. The strain was heterofermentative, non-spore-forming and rod-shaped. It produced dl-lactic acid. Based on 16S rRNA gene sequence similarity, this strain was closely related to Lactobacillus lindneri LMG 14528T (96.8 %), Lactobacillus sanfranciscensis NRIC 1548T (95.4 %) and Lactobacillus florum NRIC 0771T (95.2 %), respectively. In addition, the pheS gene sequence of strain Ru20-1T was closely related to those of L. sanfranciscensis NRIC 1548T (92.0 %), L. lindneri LMG 14528T (89.0 %) and L. florum NRIC 0771T(85.0 %). Phylogenetic analysis indicated that strain Ru20-1T was clearly separated from closely related species of the genus Lactobacillus. The DNA G+C content of strain Ru20-1T was 47.8 mol %. The cell-wall peptidoglycan type was l-Lys-d-Asp. The major cellular fatty acids were C18 : 1ω9c, C20 : 0, C20 : 1ω9c and summed feature 7 (unknown 18.846 and/or C19 : 1ω6c and/or C19 : 0 cyclo). On the basis of the data provided, strain Ru20-1T represents a novel species of the genus Lactobacillus, for which the name Lactobacillus ixorae sp. nov. is proposed. The type strain is Ru20-1T (=LMG 29008T=NBRC 111239T=PCU 346T=TISTR 2381T).

  4. Antagonistic potential against pathogenic microorganisms and hydrogen peroxide production of indigenous lactobacilli isolated from vagina of Chinese pregnant women.

    PubMed

    Xu, Heng-Yi; Tian, Wan-Hong; Wan, Cui-Xiang; Jia, Li-Jun; Wang, Lan-Yin; Yuan, Jing; Liu, Chun-Mei; Zeng, Ming; Wei, Hua

    2008-10-01

    To investigate the indigenous lactobacilli from the vagina of pregnant women and to screen the isolates with antagonistic potential against pathogenic microorganisms. The strains were isolated from pregnant women's vagina and identified using the API50CH system. The ability of the isolates to produce hydrogen peroxide was analyzed semi-quantitatively using the TMB-HRP-MRS agar. The antagonistic effects of the isolates on pathogenic microorganisms were determined with a double layer agar plate. One hundred and three lactobacilli strains were isolated from 60 samples of vaginal secretion from healthy pregnant women. Among them, 78 strains could produce hydrogen peroxide, in which 68%, 80%, 80%, and 88% had antagonistic effects against Candida albicans CMCC98001, Staphylococcus aureus CMCC26003, Escherichia coli CMCC44113, and Pseudomonas aeruginosa CMCC10110, respectively. The recovery of hydrogen peroxide-producing lactobacilli decreases with the increasing pregnant age and time. The most commonly isolated species from vagina of Chinese pregnant women are Lactobacillus acidophilus and Lactobacillus crispatus. Most of L. acidophilus and L. crispatus produce a high H2O2 level.

  5. Antimicrobial activity of Lactobacillus salivarius and Lactobacillus fermentum against Staphylococcus aureus.

    PubMed

    Kang, Mi-Sun; Lim, Hae-Soon; Oh, Jong-Suk; Lim, You-Jin; Wuertz-Kozak, Karin; Harro, Janette M; Shirtliff, Mark E; Achermann, Yvonne

    2017-03-01

    The increasing prevalence of methicillin-resistant Staphylococcus aureus has become a major public health threat. While lactobacilli were recently found useful in combating various pathogens, limited data exist on their therapeutic potential for S. aureus infections. The aim of this study was to determine whether Lactobacillus salivarius was able to produce bactericidal activities against S. aureus and to determine whether the inhibition was due to a generalized reduction in pH or due to secreted Lactobacillus product(s). We found an 8.6-log10 reduction of planktonic and a 6.3-log10 reduction of biofilm S. aureus. In contrast, the previously described anti-staphylococcal effects of L. fermentum only caused a 4.0-log10 reduction in planktonic S. aureus cells, with no effect on biofilm S. aureus cells. Killing of S. aureus was partially pH dependent, but independent of nutrient depletion. Cell-free supernatant that was pH neutralized and heat inactivated or proteinase K treated had significantly reduced killing of L. salivarius than with pH-neutralized supernatant alone. Proteomic analysis of the L. salivarius secretome identified a total of five secreted proteins including a LysM-containing peptidoglycan binding protein and a protein peptidase M23B. These proteins may represent potential novel anti-staphylococcal agents that could be effective against S. aureus biofilms. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  6. Bacterial identification of the vaginal microbiota in Ecuadorian pregnant teenagers: an exploratory analysis

    PubMed Central

    Salinas, Ana María; Osorio, Verónica Gabriela; Endara, Pablo Francisco; Salazar, Eduardo Ramiro; Vasco, Gabriela Piedad; Vivero, Sandra Guadalupe

    2018-01-01

    .7%) were found in high prevalence. Moreover, the most predominant bacterial Lactobacillus species found in the pregnant teenagers’ vaginal tract were L. crispatus (92.6%), L. iners (89.5%) and L. acidophilus (87.4%). In addition, the average vaginal pH measured in the study population was 5.2, and high pH was associated with the presence of the three-anaerobic species (p = 0.001). Finally, L. jensenii’s presence in the study decreased in 72% the occupation of the three anaerobes. Discussion This work identified a high pH as a risk factor for BV anaerobes’ presence in adolescent pregnant women. Moreover, this study identified L. crispatus, L. iners and L. acidophilus to be the most abundant species in our study population. From all fastidious anaerobes analyzed in this study, A. vaginae was present in all pregnant teenagers. To conclude, L. jensenii could be a potential healthy vaginal microbiota candidate in pregnant teenagers and should be further analyzed in future studies. PMID:29492333

  7. 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 β-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 β-glucosidase produced hydrolyzed 5-bromo-4-chloro-3-indolyl-β-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 β-3,4-dihydroxyphenylethanol. Images PMID:16349442

  8. Hydrolysis of Oleuropein by Lactobacillus plantarum Strains Associated with Olive Fermentation.

    PubMed

    Ciafardini, G; Marsilio, V; Lanza, B; Pozzi, N

    1994-11-01

    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 beta-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 beta-glucosidase produced hydrolyzed 5-bromo-4-chloro-3-indolyl-beta-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 degrees 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 beta-3,4-dihydroxyphenylethanol.

  9. Effect of ultrasound on lactic acid production by Lactobacillus strains in date (Phoenix dactylifera var. Kabkab) syrup.

    PubMed

    Hashemi, Seyed Mohammad Bagher; Mousavi Khaneghah, Amin; Saraiva, Jorge A; Jambrak, Anet Režek; Barba, Francisco J; Mota, Maria J

    2018-03-01

    Date syrup is rich in fermentable sugars and may be used as a substrate for different microbial fermentations, including lactic acid fermentation processes. The beneficial effects of ultrasounds (US) on bioprocesses have been reported for several microorganisms, due to the enhancement of cell growth, as well as improvements in yields and productivities. Therefore, US treatments (30 kHz, 100 W, 10-30 min) were applied to two lactobacilli (Lactobacillus helveticus PTCC 1332 and Lactobacillus acidophilus PTCC 1643), during fermentation using date syrup as substrate. The effects on lactic acid fermentation were evaluated by analyzing cell growth (dry cell weight and viable cell count), substrate consumption (quantification of glucose and fructose), and product formation (quantification of lactic acid) over time. The effects of US were also evaluated on cell membrane permeability. Both lactobacilli were able to grow well on date syrup without the need for addition of further ingredients. The US effects were highly dependent on treatment duration: treatments of 10- and 20-min stimulated lactobacilli growth, while the treatment extension to 30 min negatively affected cell growth. Similarly, the 10- and 20-min treatments increased sugar consumption and lactic acid production, contrarily to the 30-min treatment. All US treatments increased cell membrane permeability, with a more pronounced effect at more extended treatments. The results of this work showed that application of appropriate US treatments could be a useful tool for stimulation of lactic acid production from date syrup, as well as for other fermentative processes that use date syrup as substrate.

  10. Microbial transformation of ginsenoside Rb1 to compound K by Lactobacillus paralimentarius.

    PubMed

    Quan, Lin-Hu; Kim, Yeon-Ju; Li, Guan Hao; Choi, Kwang-Tea; Yang, Deok-Chun

    2013-06-01

    In this study, the major ginsenoside Rb1 was transformed into the more pharmacologically active minor compound K by food grade Lactobacillus paralimentarius LH4, which was isolated from kimchi, a traditional Korean fermented food. The enzymatic reaction was analyzed by TLC, HPLC, and NMR. Using the cell-free enzyme of Lactobacillus paralimentarius LH4 at optimal conditions for 30 °C at pH 6.0, 1.0 mg ml(-1) ginsenoside Rb1 was transformed into 0.52 mg ml(-1) compound K within 72 h, with a corresponding molar conversion yield of 88 %. The cell-free enzyme hydrolyzed the two glucose moieties attached to the C-3 position and the outer glucose moiety attached to the C-20 position of the ginsenoside Rb1. The cell-free enzyme hydrolyzed the ginsenoside Rb1 along the following pathway: ginsenoside Rb1 → gypenoside XVII and ginsenoside Rd → ginsenoside F2 → compound K. Our results indicate that Lactobacillus paralimentarius LH4 has the potential to be applied for the preparation of compound K in the food industry.

  11. O2(b1Σg+) Quenching by O2, CO2, H2O, and N2 at Temperatures of 300-800 K.

    PubMed

    Zagidullin, M V; Khvatov, N A; Medvedkov, I A; Tolstov, G I; Mebel, A M; Heaven, M C; Azyazov, V N

    2017-10-05

    Rate constants for the removal of O 2 (b 1 Σ g + ) by collisions with O 2 , N 2 , CO 2 , and H 2 O have been determined over the temperature range from 297 to 800 K. O 2 (b 1 Σ g + ) was excited by pulses from a tunable dye laser, and the deactivation kinetics were followed by observing the temporal behavior of the b 1 Σ g + -X 3 Σ g - fluorescence. The removal rate constants for CO 2 , N 2 , and H 2 O were not strongly dependent on temperature and could be represented by the expressions k CO2 = (1.18 ± 0.05) × 10 -17 × T 1.5 × exp[Formula: see text], k N2 = (8 ± 0.3) × 10 -20 × T 1.5 × exp[Formula: see text], and k H2O = (1.27 ± 0.08) × 10 -16 × T 1.5 × exp[Formula: see text] cm 3 molecule -1 s -1 . Rate constants for O 2 (b 1 Σ g + ) removal by O 2 (X), being orders of magnitude lower, demonstrated a sharp increase with temperature, represented by the fitted expression k O2 = (7.4 ± 0.8) × 10 -17 × T 0.5 × exp[Formula: see text] cm 3 molecule -1 s -1 . All of the rate constants measured at room temperature were found to be in good agreement with previously reported values.

  12. Prevention of Escherichia coli infection in broiler chickens with Lactobacillus plantarum B1.

    PubMed

    Wang, S; Peng, Q; Jia, H M; Zeng, X F; Zhu, J L; Hou, C L; Liu, X T; Yang, F J; Qiao, S Y

    2017-08-01

    Two studies were performed to assess the efficacy of Lactobacillus plantarum B1 in prevention of pathogenic Escherichia coli K88 gastrointestinal infection in broilers. In an in vitro study, L. plantarum B1 showed resistance to acid and bile and inhibited the growth of E. coli K88. Additionally, L. plantarum B1 exhibited high ability to adhere to broiler embryo ileal epithelium. In an animal trial, 240 broilers at 1 d of age were randomly assigned to one of 4 treatment arms: negative control (NC) broilers fed a basal diet and not challenged; positive control (PC) broilers fed a basal diet and challenged with E. coli K88; L. plantarum (LP) treatment broilers fed a basal diet containing 2 × 109 cfu/kg L. plantarum B1 and challenged with E. coli K88; and antibiotic treatment (Anti) broilers fed a basal diet supplemented with colistin sulfate (20 mg/kg) and challenged with E. coli K88. Broilers fed L. plantarum B1 had greater (P ≤ 0.05) BW than those in the PC treatment on d 14 and 28. Dietary L. plantarum B1 decreased (P < 0.05) E. coli counts in the cecal contents on d 10 and 14, and increased (P < 0.05) cecal lactic acid bacteria (LAB) on d 8, 10, 14, and 28 compared with the PC treatment. Dietary supplementation of L. plantarum B1 increased (P < 0.05) the ileal mucosal secretory IgA concentration and reduced (P < 0.05) IL-2, IL-4, IFN-γ, and tumor necrosis factor-α levels in the ileum. Overall, these results suggest dietary supplementation of L. plantarum B1 promotes growth performance, lowers cecal E. coli counts, and increases the population of cecal LAB, as well as improves intestinal mucosal immunity in E. coli K88-challenged broilers. © 2017 Poultry Science Association Inc.

  13. Novel antibacterial polypeptide laparaxin produced by Lactobacillus paracasei strain NRRL B-50314 via fermentation

    USDA-ARS?s Scientific Manuscript database

    This study reports the production and characterization of a novel antibacterial polypeptide, designated laparaxin, which is secreted by Lactobacillus paracasei NRRL B-50314. Crude laparaxin has antibacterial activity against a wide variety of Gram-positive bacteria, including: lactic acid bacteria ...

  14. Design, synthesis and anticonvulsant activity of some new 6,8-halo-substituted-2h-[1,2,4]triazino[5,6-b]indole-3(5h)-one/-thione and 6,8-halo-substituted 5-methyl-2h-[1,2,4]triazino[5,6-b]indol-3(5h)-one/-thione

    PubMed Central

    Kumar, Rajeev; Singh, Tejendra; Singh, Hariram; Jain, Sandeep; Roy, R. K.

    2014-01-01

    A new series of 6,8-halo-substituted-2H-[1,2,4]triazino[5,6-b]indole-3(5H)-one/-thione and 6,8-halo-substituted 5-methyl-2H-[1,2,4]triazino[5,6-b]indol-3(5H)-one/-thione (5a-5l) were designed and synthesized keeping in view of the structural requirement of pharmacophore. The above compounds were characterized by thin layer chromatography and spectral analysis. Anticonvulsant activity of the synthesized compounds was evaluated by the maximal electroshock (MES) test. Neurotoxicity and CNS depressant effects were evaluated by the rotarod motor impairment and Porsolt’s force swim tests, respectively. A computational study was carried out, for calculation of pharmacophore pattern, prediction of pharmacokinetic properties and toxicity properties. The above study revealed that the compounds 8-chloro-2H-[1,2,4]triazino[5,6-b]indol-3(5H)-one (5e), 6,8-dibromo-2H-[1,2,4]triazino[5,6-b]indol-3(5H)-one (5i) and 6,8-dibromo-5-methyl-2H-[1,2,4]triazino[5,6-b]indol-3(5H)-one (5k) possess excellent anticonvulsant activity in the series with little CNS depressant effect and no neurotoxicity as compared to standard drugs phenytoin and carbamazepine. PMID:26417257

  15. Characterization of glutamate decarboxylase from Lactobacillus plantarum and its C-terminal function for the pH dependence of activity.

    PubMed

    Shin, Sun-Mi; Kim, Hana; Joo, Yunhye; Lee, Sang-Jae; Lee, Yong-Jik; Lee, Sang Jun; Lee, Dong-Woo

    2014-12-17

    The gadB gene encoding glutamate decarboxylase (GAD) from Lactobacillus plantarum was cloned and expressed in Escherichia coli. The recombinant enzyme exhibited maximal activity at 40 °C and pH 5.0. The 3D model structure of L. plantarum GAD proposed that its C-terminal region (Ile454-Thr468) may play an important role in the pH dependence of catalysis. Accordingly, C-terminally truncated (Δ3 and Δ11 residues) mutants were generated and their enzyme activities compared with that of the wild-type enzyme at different pH values. Unlike the wild-type GAD, the mutants showed pronounced catalytic activity in a broad pH range of 4.0-8.0, suggesting that the C-terminal region is involved in the pH dependence of GAD activity. Therefore, this study may provide effective target regions for engineering pH dependence of GAD activity, thereby meeting industrial demands for the production of γ-aminobutyrate in a broad range of pH values.

  16. 20 CFR 655.1305 - Assurances and obligations of H-2A employers.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... 20 Employees' Benefits 3 2014-04-01 2014-04-01 false Assurances and obligations of H-2A employers... Temporary Agricultural Employment in the United States (H-2A Workers) § 655.1305 Assurances and obligations of H-2A employers. An employer seeking to employ H-2A workers must attest as part of the Application...

  17. Lactobacillus

    MedlinePlus

    ... eye symptoms. Preventing diarrhea caused by antibiotics. Taking probiotics products containing lactobacillus strains helps prevent diarrhea caused ... the first 1-2 years of life, lactobacillus probiotics can reduce the chance of the child developing ...

  18. Lactobacillus species isolated from vaginal secretions of healthy and bacterial vaginosis-intermediate Mexican women: a prospective study

    PubMed Central

    2013-01-01

    Background Lactobacillus jensenii, L. iners, L. crispatus and L. gasseri are the most frequently occurring lactobacilli in the vagina. However, the native species vary widely according to the studied population. The present study was performed to genetically determine the identity of Lactobacillus strains present in the vaginal discharge of healthy and bacterial vaginosis (BV) intermediate Mexican women. Methods In a prospective study, 31 strains preliminarily identified as Lactobacillus species were isolated from 21 samples collected from 105 non-pregnant Mexican women. The samples were classified into groups according to the Nugent score criteria proposed for detection of BV: normal (N), intermediate (I) and bacterial vaginosis (BV). We examined the isolates using culture-based methods as well as molecular analysis of the V1–V3 regions of the 16S rRNA gene. Enterobacterial repetitive intergenic consensus (ERIC) sequence analysis was performed to reject clones. Results Clinical isolates (25/31) were classified into four groups based on sequencing and analysis of the 16S rRNA gene: L. acidophilus (14/25), L. reuteri (6/25), L. casei (4/25) and L. buchneri (1/25). The remaining six isolates were presumptively identified as Enterococcus species. Within the L. acidophilus group, L. gasseri was the most frequently isolated species, followed by L. jensenii and L. crispatus. L. fermentum, L. rhamnosus and L. brevis were also isolated, and were placed in the L. reuteri, L. casei and L. buchneri groups, respectively. ERIC profile analysis showed intraspecific variability amongst the L. gasseri and L. fermentum species. Conclusions These findings agree with previous studies showing that L. crispatus, L. gasseri and L. jensenii are consistently present in the healthy vaginal ecosystem. Additional species or phylotypes were detected in the vaginal microbiota of the non-pregnant Mexican (Hispanic-mestizo) population, and thus, these results further our understanding of

  19. The D/H Ratio in Interstellar Gas towards G191-B2B from STIS Echelle Observations

    NASA Astrophysics Data System (ADS)

    Sahu, M. S.; Landsman, W. B.; Bruhweiler, F. C.; Gull, T. R.; Bowers, C. A.; Lindler, D.; Feggans, K.; Barstow, M. A.; Hubeny, I.; Holberg, J. B.

    1999-05-01

    We present STIS echelle observations of interstellar D i and H i Lyα and N i (1199.5, 1200.2 and 1200.7 Angstroms), C ii 1334.5 Angstroms, C(*) ii 1335.7 Angstroms, O i 1302 Angstroms, Si ii (1190, 1193, 1260, 1304 and 1526 Angstroms), Si iii 1206.5 Angstroms, Al ii 1670.8 Angstroms, S ii 1259.5 Angstroms and Fe ii 1608.5 Angstroms in the line of sight to the nearby (69 pc) hot, white dwarf (WD) G191-B2B. Compared to the GHRS study of G191-B2B by Vidal-Madjar et al. 1998 (VM98), the STIS E140H spectra have a higher velocity resolution (3 km s(-1) ), better S/N (between 20 to 50) and broader wavelength coverage (1150 to 1700 Angstroms). We use the Barstow & Hubeny stratified non-LTE model atmosphere calculations which include the effects of line-blanketing from more than 9x10(6) atomic transitions (mainly Ni and Fe), both to determine the NLTE shape of the stellar Lyalpha profile and to estimate the contamination of the interstellar lines by WD photospheric lines. The interstellar N i 1200.7 Angstroms, Si ii 1193 & 1304 Angstroms and Fe ii lines show no contamination by WD photospheric lines and are given more weight in our analysis. VM98 reported three components while we detect only two velocity components in all the interstellar species observed: one at ~ 8.5 km s(-1) and one at ~ 19.3 km s(-1) which we identify as the LIC component. Using the NLTE stellar Lyα profile and a total column density of N(H i) ~ 2 x 10(18) cm(-2) for both components (consistent with EUVE observations), we derive confidence contours. We find the D/H ratio with 2sigma confidence limits to lie within 1.77+/-0.2x10(-5) . This value is consistent with the value of (D/H)LIC = 1.6+/-0.1x10(-5) determined towards Capella (Linsky et al. 1995). The STIS data provide no evidence for local or cloud-to-cloud variation in the D/H ratio as suggested by VM98. Re-analysis of the GHRS data and comparison to the STIS data is in progress.

  20. Bactericidal activity of culture fluid components of Lactobacillus fermentum strain 90 TS-4 (21) clone 3, and their capacity to modulate adhesion of Candida albicans yeast-like fungi to vaginal epithelial cells.

    PubMed

    Anokhina, I V; Kravtsov, E G; Protsenko, A V; Yashina, N V; Yermolaev, A V; Chesnokova, V L; Dalin, M V

    2007-03-01

    Antagonistic activities of L. fermentum strain 90 TS-4 (21), L. casei ATCC 27216, and L. acidophilus ATCC 4356 and bactericidal activity of lactobacillus culture fluid towards E. coli strain K12, S. aureus, and S. epidermidis test cultures were studied. The bactericidal effect of L. fermentum strain 90 TS-4 (21) clone 3 culture fluid preparation (pH 6.0) on the test cultures was dose-dependent. Adhesion of C. albicans yeast-like fungi to vaginal epitheliocytes was more pronounced for strains isolated from women with asymptomatic infection than for strains isolated from women with manifest forms. L. fermentum strain 90 TS-4 (21) clone 3 culture fluid preparation modulated adhesion of yeast-like fungi only if the fungal strain was initially highly adherent.

  1. Lactobacillus GG prevents recurrence of colitis in HLA-B27 transgenic rats after antibiotic treatment

    PubMed Central

    Dieleman, L A; Goerres, M S; Arends, A; Sprengers, D; Torrice, C; Hoentjen, F; Grenther, W B; Sartor, R B

    2003-01-01

    Background and aims: Bacteroides vulgatus induces colitis in gnotobiotic HLA-B27 transgenic (TG) rats while broad spectrum antibiotics prevent and treat colitis in specific pathogen free (SPF) TG rats although disease recurs after treatment ends. Lactobacilli treat human pouchitis and experimental colitis. We investigated if Lactobacillus rhamnosus GG (L GG) can prevent colitis in TG rats monoassociated with B vulgatus and if L GG or Lactobacillus plantarum 299v (LP 299v) can treat established colitis in SPF TG rats and prevent recurrent disease after antibiotics were stopped. Methods: Germfree B27 TG rats were monoassociated with B vulgatus for four weeks following two weeks of colonisation with L GG or no bacteria. SPF B27 TG rats received oral vancomycin and imipenem for two weeks, or water alone, followed by four weeks of treatment with oral L GG, LP 299v, or water only. Disease activity was quantified by blinded gross and histological scores, caecal myeloperoxidase (MPO) activity, and levels of interleukin (IL)-1β, tumour necrosis factor (TNF), transforming growth factor β, and IL-10. Results: L GG did not prevent colitis in B vulgatus co-associated TG rats or treat established disease in SPF rats. However, L GG but not LP 299v prevented colitis relapse in antibiotic treated rats with reduced gross and histological scores, caecal MPO, IL-1β, and TNF whereas caecal IL-10 was increased. Conclusions: L GG does not prevent colitis in gnotobiotic TG rats or treat established disease in SPF rats, but is superior to LP 299v in the prevention of recurrent colitis. These studies suggest that antibiotics and probiotic agents provide synergistic therapeutic effects, perhaps mediated by altered immunomodulation with selective activity of different lactobacillus species. PMID:12584218

  2. Lactobacillus GG prevents recurrence of colitis in HLA-B27 transgenic rats after antibiotic treatment.

    PubMed

    Dieleman, L A; Goerres, M S; Arends, A; Sprengers, D; Torrice, C; Hoentjen, F; Grenther, W B; Sartor, R B

    2003-03-01

    Bacteroides vulgatus induces colitis in gnotobiotic HLA-B27 transgenic (TG) rats while broad spectrum antibiotics prevent and treat colitis in specific pathogen free (SPF) TG rats although disease recurs after treatment ends. Lactobacilli treat human pouchitis and experimental colitis. We investigated if Lactobacillus rhamnosus GG (L GG) can prevent colitis in TG rats monoassociated with B vulgatus and if L GG or Lactobacillus plantarum 299v (LP 299v) can treat established colitis in SPF TG rats and prevent recurrent disease after antibiotics were stopped. Germfree B27 TG rats were monoassociated with B vulgatus for four weeks following two weeks of colonisation with L GG or no bacteria. SPF B27 TG rats received oral vancomycin and imipenem for two weeks, or water alone, followed by four weeks of treatment with oral L GG, LP 299v, or water only. Disease activity was quantified by blinded gross and histological scores, caecal myeloperoxidase (MPO) activity, and levels of interleukin (IL)-1 beta, tumour necrosis factor (TNF), transforming growth factor beta, and IL-10. L GG did not prevent colitis in B vulgatus co-associated TG rats or treat established disease in SPF rats. However, L GG but not LP 299v prevented colitis relapse in antibiotic treated rats with reduced gross and histological scores, caecal MPO, IL-1 beta, and TNF whereas caecal IL-10 was increased. L GG does not prevent colitis in gnotobiotic TG rats or treat established disease in SPF rats, but is superior to LP 299v in the prevention of recurrent colitis. These studies suggest that antibiotics and probiotic agents provide synergistic therapeutic effects, perhaps mediated by altered immunomodulation with selective activity of different lactobacillus species.

  3. Modeling Lactic Fermentation of Gowé Using Lactobacillus Starter Culture.

    PubMed

    de J C Munanga, Bettencourt; Loiseau, Gérard; Grabulos, Joël; Mestres, Christian

    2016-12-01

    A global model of the lactic fermentation step of gowé was developed by assembling blocks hosting models for bacterial growth, lactic acid production, and the drop of pH during fermentation. Commercial strains of Lactobacillus brevis and of Lactobacillus plantarum were used; their growth was modeled using Rosso's primary model and the gamma concept as a secondary model. The optimum values of pH and temperature were 8.3 ± 0.3, 44.6 ± 1.2 °C and 8.3 ± 0.3, 3.2 ± 37.1 °C with μ max values of 1.8 ± 0.2 and 1.4 ± 0.1 for L. brevis and L. plantarum respectively. The minimum inhibitory concentration of undissociated lactic acid was 23.7 mM and 35.6 mM for L. brevis and L. plantarum , respectively. The yield of lactic acid was five times higher for L. plantarum than for L. brevis , with a yield of glucose conversion to lactic acid close to 2.0 for the former and 0.8 for the latter. A model was developed to predict the pH drop during gowé fermentation. The global model was partially validated during manufacturing of gowé. The global model could be a tool to aid in the choice of suitable starters and to determine the conditions for the use of the starter.

  4. Comparison of the antibacterial activity of different self-etching primers and adhesives.

    PubMed

    Korkmaz, Yonca; Ozalp, Meral; Attar, Nuray

    2008-11-01

    The aim of this study was to evaluate the antibacterial effects of different one-step and two-step self-etching primer/adhesives on Streptococcus mutans (S. mutans), Lactobacillus casei (L. casei), and Lactobacillus acidophilus (L. acidophilus). The antibacterial effects of Clearfil Protect Bond Primer and Bonding agent; AdheSE Primer and Bonding agent; Adper Prompt L-Pop; Futurabond NR; Clearfil Tri S Bond; and Cervitec (positive control, 1% chlorhexidine varnish) were tested against standard strains of S. mutans, L. Casei, and L. acidophilus using the disk diffusion method. Standard filter paper disks (n=5) impregnated with 20 microL of each material were prepared. After incubation at 37 masculineC for 48 hours in a 5-10% CO2 atmosphere, the diameter of inhibition zones were measured in millimeters. Data were analyzed using one way analysis of variance (ANOVA) and multivariate analysis of variance (MANOVA). Duncan's Multiple Range Test was used for pairwise comparison. The size of inhibition zones produced by primer/adhesives varied among the brands. AdheSE Primer: S. mutans (20.6+/-1.51); L. casei (14.8+/-1.78); L. acidophilus (11.4+/-0.54). Adper Prompt L-Pop: S. mutans (19.6+/-1.51); L. casei (13.8+/-1.64); L. acidophilus (13.8+/-1.09). Cervitec: S. mutans (23+/-0.00); L. casei (27+/-0.70); L. acidophilus (22.4+/-0.54). Clearfil Protect Bond Primer: S. mutans (17+/-0.00); L. casei (17.6+/-0.54); L. acidophilus (22.4+/-0.54). Futurabond NR was found effective only against S. mutans (14.6+/-1.67). Of all the materials tested, AdheSE Bonding agent, Clearfil Protect Bond Bonding agent, and Clearfil Tri S Bond exhibited no inhibition zone (-) for all bacteria tested. Among the adhesives tested Clearafil Protect Bond Primer based upon monomer methacryloyloxydodecylpyridiniium bromide (MDPB) was found to be the most potent material against L. acidophilus and L. casei. AdheSE Primer and Adper Prompt L-Pop are highly effective against S. mutans. Compared with other

  5. Co-administration of Bacillus subtilis RJGP16 and Lactobacillus salivarius B1 strongly enhances the intestinal mucosal immunity of piglets.

    PubMed

    Deng, Jun; Li, Yunfeng; Zhang, Jinhua; Yang, Qian

    2013-02-01

    Probiotics, including Bacillus spp. and Lactobacillus, are potential replacements for low dose in-feed antibiotics for pig. This study aimed to evaluate the effect of the co-administration of Bacillus subtilis RJGP16 and Lactobacillus salivarius B1 as potential probiotics to stimulate local immune responses. Thirty two newborn piglets were divided into four groups and orally administrated with different combination of probiotics (none; RJGP16; B1; RJGP16 and B1) at the age of 0, 7 and 11 days. We analysed the parameters of the mucosal immunity of piglets a week after weaning. Our results showed that the gene expression of interleukin (IL)-6 in the duodenum and ileum, porcine beta-defensins (pBD)-2 in the duodenum were significantly increased (p<0.01) with co-administration of the RJGP16 and B1. Also the expression and release of TLR-2 and the number of immunoglobulin (Ig) A producing cells were increased (p<0.01). The results demonstrate that the co-administration of the two bacteria stimulate a more intense mucosal immunity than the administration of each bacterium alone. Copyright © 2012 Elsevier Ltd. All rights reserved.

  6. Broad N2H+ Emission toward the Protostellar Shock L1157-B1

    NASA Astrophysics Data System (ADS)

    Codella, C.; Viti, S.; Ceccarelli, C.; Lefloch, B.; Benedettini, M.; Busquet, G.; Caselli, P.; Fontani, F.; Gómez-Ruiz, A.; Podio, L.; Vasta, M.

    2013-10-01

    We present the first detection of N2H+ toward a low-mass protostellar outflow, namely, the L1157-B1 shock, at ~0.1 pc from the protostellar cocoon. The detection was obtained with the IRAM 30 m antenna. We observed emission at 93 GHz due to the J = 1-0 hyperfine lines. Analysis of this emission coupled with HIFI CHESS multiline CO observations leads to the conclusion that the observed N2H+(1-0) line originated from the dense (>=105 cm-3) gas associated with the large (20''-25'') cavities opened by the protostellar wind. We find an N2H+ column density of a few 1012 cm-2 corresponding to an abundance of (2-8) × 10-9. The N2H+ abundance can be matched by a model of quiescent gas evolved for more than 104 yr, i.e., for more than the shock kinematical age (sime2000 yr). Modeling of C-shocks confirms that the abundance of N2H+ is not increased by the passage of the shock. In summary, N2H+ is a fossil record of the pre-shock gas, formed when the density of the gas was around 104 cm-3, and then further compressed and accelerated by the shock.

  7. The Mediator subunit MED23 couples H2B mono-ubiquitination to transcriptional control and cell fate determination.

    PubMed

    Yao, Xiao; Tang, Zhanyun; Fu, Xing; Yin, Jingwen; Liang, Yan; Li, Chonghui; Li, Huayun; Tian, Qing; Roeder, Robert G; Wang, Gang

    2015-12-02

    The Mediator complex orchestrates multiple transcription factors with the Pol II apparatus for precise transcriptional control. However, its interplay with the surrounding chromatin remains poorly understood. Here, we analyze differential histone modifications between WT and MED23(-/-) (KO) cells and identify H2B mono-ubiquitination at lysine 120 (H2Bub) as a MED23-dependent histone modification. Using tandem affinity purification and mass spectrometry, we find that MED23 associates with the RNF20/40 complex, the enzyme for H2Bub, and show that this association is critical for the recruitment of RNF20/40 to chromatin. In a cell-free system, Mediator directly and substantially increases H2Bub on recombinant chromatin through its cooperation with RNF20/40 and the PAF complex. Integrative genome-wide analyses show that MED23 depletion specifically reduces H2Bub on a subset of MED23-controlled genes. Importantly, MED23-coupled H2Bub levels are oppositely regulated during myogenesis and lung carcinogenesis. In sum, these results establish a mechanistic link between the Mediator complex and a critical chromatin modification in coordinating transcription with cell growth and differentiation. © 2015 The Authors.

  8. A Novel Dimeric Ni-Substituted beta-Keggin Silicotungstate: Structure and Magnetic Properties of K(12)[{beta-SiNi(2)W(10)O(36)(OH)(2)(H(2)O)}(2)].20H(2)O.

    PubMed

    Kortz, Ulrich; Jeannin, Yves P.; Tézé, André; Hervé, Gilbert; Isber, Samih

    1999-08-09

    The novel dimeric polyoxometalate [{beta-SiNi(2)W(10)O(36)(OH)(2)(H(2)O)}(2)](12)(-) (1) has been synthesized and characterized by IR spectroscopy, polarography, elemental analysis, thermogravimetric analysis, and magnetic measurements. An X-ray single-crystal analysis was carried out on K(12)[{beta-SiNi(2)W(10)O(36)(OH)(2)(H(2)O)}(2)].20H(2)O, which crystallizes in the monoclinic system, space group P2(1)/n, with a = 13.701(4) Å, b = 24.448(11) Å, c = 13.995(5) Å, beta = 99.62(3) degrees, and Z = 4. The anion consists of two [beta-SiNi(2)W(10)O(36)(OH)(2)(H(2)O)] Keggin moieties linked via two OH bridging groups, leading to a planar Ni(2)(OH)(2) unit. The two half-units are related by an inversion center and each contain one Ni atom in the rotated triad. The formation of the new anion involves insertion, isomerization, and dimerization. Magnetic measurements show that the central Ni(4) unit exhibits ferromagnetic (J' = 4.14 cm(-)(1)) as well as weak antiferromagnetic (J = -0.65 cm(-)(1)) Ni-Ni exchange interactions.

  9. Purification and genetic characterisation of the novel bacteriocin LS2 produced by the human oral strain Lactobacillus salivarius BGHO1.

    PubMed

    Busarcevic, Milos; Dalgalarrondo, Michèle

    2012-08-01

    The aim of this study was to investigate the antimicrobial potential of Lactobacillus salivarius BGHO1, a human oral strain with probiotic characteristics and a broad inhibitory spectrum both against Gram-positive and Gram-negative pathogens. Here we present the bacteriocin LS2, an extremely pH- and heat-stable peptide with antilisterial activity. LS2 is a novel member of the class IId bacteriocins, unique among all currently characterised bacteriocins. It is somewhat similar to putative bacteriocins from several oral streptococci, including the cariogenic Streptococcus mutans. LS2 is a 41-amino-acid, highly hydrophobic cationic peptide of 4115.1Da that is sensitive to proteolytic enzymes. LS2 was purified from cells of strain BGHO1 by solvent extraction and reverse-phase chromatography. Mass spectrometry was used to determine the molecular mass of the purified peptide. N-terminal amino acid sequencing enabled identification of the LS2 structural gene bacls2 by a reverse genetics approach. Downstream of the bacls2 gene, two bacteriocin-like genes were found, named blp1a and blp1b, and one putative bacteriocin immunity gene named bimlp. We also present the identification of the 242-kb megaplasmid pMPHO1 by pulsed-field gel electrophoresis, which harbours the genes bacls2, blp1a, blp1b and bimlp. Two peptides with antimicrobial activity, whose approximate sizes corresponded to those of blp1a and blp1b, were identified only after culturing strain BGHO1 in a chemically defined medium. This study demonstrated the capacity of Lactobacillus salivarius BGHO1 to produce multiple bacteriocins and further established this strain as a promising probiotic candidate. Copyright © 2012 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

  10. 20 CFR 655.700 - What statutory provisions govern the employment of H-1B, H-1B1, and E-3 nonimmigrants and how do...

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... employment of H-1B, H-1B1, and E-3 nonimmigrants and how do employers apply for H-1B, H-1B1, and E-3 visas... Requirements for Employers Seeking To Employ Nonimmigrants on H-1b Visas in Specialty Occupations and as Fashion Models, and Requirements for Employers Seeking To Employ Nonimmigrants on H-1b1 and E-3 Visas in...

  11. 20 CFR 655.700 - What statutory provisions govern the employment of H-1B, H-1B1, and E-3 nonimmigrants and how do...

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... employment of H-1B, H-1B1, and E-3 nonimmigrants and how do employers apply for H-1B, H-1B1, and E-3 visas... Requirements for Employers Seeking To Employ Nonimmigrants on H-1b Visas in Specialty Occupations and as Fashion Models, and Requirements for Employers Seeking To Employ Nonimmigrants on H-1b1 and E-3 Visas in...

  12. 20 CFR 655.700 - What statutory provisions govern the employment of H-1B, H-1B1, and E-3 nonimmigrants and how do...

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... employment of H-1B, H-1B1, and E-3 nonimmigrants and how do employers apply for H-1B, H-1B1, and E-3 visas... Requirements for Employers Seeking To Employ Nonimmigrants on H-1b Visas in Specialty Occupations and as Fashion Models, and Requirements for Employers Seeking To Employ Nonimmigrants on H-1b1 and E-3 Visas in...

  13. 20 CFR 655.700 - What statutory provisions govern the employment of H-1B, H-1B1, and E-3 nonimmigrants and how do...

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... employment of H-1B, H-1B1, and E-3 nonimmigrants and how do employers apply for H-1B, H-1B1, and E-3 visas... Requirements for Employers Seeking To Employ Nonimmigrants on H-1b Visas in Specialty Occupations and as Fashion Models, and Requirements for Employers Seeking To Employ Nonimmigrants on H-1b1 and E-3 Visas in...

  14. 20 CFR 655.700 - What statutory provisions govern the employment of H-1B, H-1B1, and E-3 nonimmigrants and how do...

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... employment of H-1B, H-1B1, and E-3 nonimmigrants and how do employers apply for H-1B, H-1B1, and E-3 visas... Requirements for Employers Seeking To Employ Nonimmigrants on H-1b Visas in Specialty Occupations and as Fashion Models, and Requirements for Employers Seeking To Employ Nonimmigrants on H-1b1 and E-3 Visas in...

  15. Flavoring Production in Kamut®, Quinoa and Wheat Doughs Fermented by Lactobacillus paracasei, Lactobacillus plantarum, and Lactobacillus brevis: A SPME-GC/MS Study.

    PubMed

    Di Renzo, Tiziana; Reale, Anna; Boscaino, Floriana; Messia, Maria C

    2018-01-01

    This study identified the odor-active compounds and the qualitative characteristics of doughs from "ancient" grains flours fermented by lactic acid bacteria. For this purpose doughs made with quinoa and Kamut® flours have been produced and inoculated with strains belonging to the species Lactobacillus paracasei, Lactobacillus plantarum and Lactobacillus brevis and compared with fermented doughs made from 100% wheat flour. The quality of the doughs was determined by assessment of pH, total titratable acidity, lactic acid bacteria growth and flavor compounds. The results showed that lactic acid bacteria used were able to grow in the different substrates reaching more than 9.0 log CFU/g after 24 h fermentation, although the best microbial growth was recorded in the doughs made with quinoa flour fermented with Lactobacillus paracasei I1. Good acidification and heterogeneous aromatic profile were recognized in all the doughs even if the volatile composition mainly derived from microbial specie. Among all the used strains, mostly Lactobacillus paracasei I1 positively contributed to the aromatic profile of the doughs, independently from flour type, producing the highest amount of different ketones such as, diacetyl, acetoin, 2,6-dimethyl-4-heptanone, 5-methyl-3-hexanone, 4-methyl-3-penten-2-one, volatile compounds highly appreciated in the bakery products for their buttery, fatty and fruity notes. So, the positive characteristic of Lactobacillus paracasei I1 to enhance the production of desired volatile compounds could make it suitable as adjunct culture starter in the bakery industry. Many differences in volatile organic compounds derived also by the type of flour used. Quinoa fermented doughs were characterized for specific nutty, roasted, acid and buttery tones derived from pyrazines, ketones and acid compounds whereas Kamut® fermented doughs were characterized for fruity, rose, green and sweet tones derived from aldehydes and ketones production. So, the use of

  16. Use of Probiotics to Control Aflatoxin Production in Peanut Grains

    PubMed Central

    da Silva, Juliana Fonseca Moreira; Peluzio, Joenes Mucci; Madeira, Jovita Eugênia Gazzinelli Cruz; Silva, Marize Oliveira; de Morais, Paula Benevides; Rosa, Carlos Augusto; Pimenta, Raphael Sanzio; Nicoli, Jacques Robert

    2015-01-01

    Probiotic microorganisms (Saccharomyces cerevisiae var. boulardii, S. cerevisiae UFMG 905, and Lactobacillus delbrueckii UFV H2b20) were evaluated as biological control agents to reduce aflatoxin and spore production by Aspergillus parasiticus IMI 242695 in peanut. Suspensions containing the probiotics alone or in combinations were tested by sprinkling on the grains followed by incubation for seven days at 25°C. All probiotic microorganisms, in live and inactivated forms, significantly reduced A. parasiticus sporulation, but the best results were obtained with live cells. The presence of probiotics also altered the color of A. parasiticus colonies but not the spore morphology. Reduction in aflatoxin production of 72.8 and 65.8% was observed for S. boulardii and S. cerevisiae, respectively, when inoculated alone. When inoculated in pairs, all probiotic combinations reduced significantly aflatoxin production, and the best reduction was obtained with S. boulardii plus L. delbrueckii (96.1%) followed by S. boulardii plus S. cerevisiae and L. delbrueckii plus S. cerevisiae (71.1 and 66.7%, resp.). All probiotics remained viable in high numbers on the grains even after 300 days. The results of the present study suggest a different use of probiotics as an alternative treatment to prevent aflatoxin production in peanut grains. PMID:26221629

  17. Use of Probiotics to Control Aflatoxin Production in Peanut Grains.

    PubMed

    da Silva, Juliana Fonseca Moreira; Peluzio, Joenes Mucci; Prado, Guilherme; Madeira, Jovita Eugênia Gazzinelli Cruz; Silva, Marize Oliveira; de Morais, Paula Benevides; Rosa, Carlos Augusto; Pimenta, Raphael Sanzio; Nicoli, Jacques Robert

    2015-01-01

    Probiotic microorganisms (Saccharomyces cerevisiae var. boulardii, S. cerevisiae UFMG 905, and Lactobacillus delbrueckii UFV H2b20) were evaluated as biological control agents to reduce aflatoxin and spore production by Aspergillus parasiticus IMI 242695 in peanut. Suspensions containing the probiotics alone or in combinations were tested by sprinkling on the grains followed by incubation for seven days at 25°C. All probiotic microorganisms, in live and inactivated forms, significantly reduced A. parasiticus sporulation, but the best results were obtained with live cells. The presence of probiotics also altered the color of A. parasiticus colonies but not the spore morphology. Reduction in aflatoxin production of 72.8 and 65.8% was observed for S. boulardii and S. cerevisiae, respectively, when inoculated alone. When inoculated in pairs, all probiotic combinations reduced significantly aflatoxin production, and the best reduction was obtained with S. boulardii plus L. delbrueckii (96.1%) followed by S. boulardii plus S. cerevisiae and L. delbrueckii plus S. cerevisiae (71.1 and 66.7%, resp.). All probiotics remained viable in high numbers on the grains even after 300 days. The results of the present study suggest a different use of probiotics as an alternative treatment to prevent aflatoxin production in peanut grains.

  18. Metallaborane reaction chemistry. A predicted and found tailored facile and reversible capture of SO2 by a B-frame-supported bimetallic: structures of [(PMe2Ph)2PtPd(phen)B10H10] and [(PMe2Ph)2Pt(SO2)Pd(phen)B10H10].

    PubMed

    Bould, Jonathan; Kennedy, John D

    2008-06-07

    The formally closo twelve-vertex {ortho-M2B10} dimetallaborane system has been predictively tailored for reversible uptake of SO2 across the metal-metal bond, as exemplified by the formation of [(PMe2Ph)2Pt(SO2)Pd(phen)B10H10] from [(PMe2Ph)2PtPd(phen)B10H10].

  19. Probiotics in foods not containing milk or milk constituents, with special reference to Lactobacillus plantarum 299v.

    PubMed

    Molin, G

    2001-02-01

    Lactic acid fermentation is the simplest and safest way of preserving food and has probably always been used by humans. Species such as Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus paracasei, Lactobacillus acidophilus, and Lactobacillus salivarius are common in the human mucosa, from the mouth to the rectum. In food, L. paracasei and L. rhamnosus are usually associated with dairy products whereas L. plantarum is found in fermented foods of plant origin. A probiotic food product containing no milk constituent was launched in Sweden in 1994. The product is a lactic acid fermented oatmeal gruel that is mixed in a fruit drink. It contains approximately 5 x 10(10) colony-forming units of L. plantarum 299v/L. The strain L. plantarum 299v originates from the human intestinal mucosa and has been shown in rats to decrease translocation, improve mucosal status, improve liver status, improve the immunologic status of the mucosa, and reduce mucosal inflammation. In humans, L. plantarum 299v can increase the concentration of carboxylic acids in feces and decrease abdominal bloating in patients with irritable bowel disease. It can also decrease fibrinogen concentrations in blood. Should probiotics be administrated through foods, the probiotic organism must remain vigorous in the food until consumption and the food must remain palatable, ie, the food carrier and the organism must suit each other. L. plantarum 299v not only affects the bacterial flora of the intestinal mucosa but may also regulate the host's immunologic defense. The mechanisms involved need to be clarified.

  20. Multiple antibacterial histone H2B proteins are expressed in tissues of American oyster.

    PubMed

    Seo, Jung-Kil; Stephenson, Jeana; Noga, Edward J

    2011-03-01

    We have previously identified a histone H2B isomer (cvH2B-1) from tissue extracts of the bivalve mollusk, the American oyster (Crassostrea virginica). In this paper, we isolate an additional three antibacterial proteins from acidified gill extract by preparative acid-urea-polyacrylamide gel electrophoresis and reversed-phase high performance liquid chromatography. Extraction of these proteins from tissue was best accomplished by briefly boiling the tissues in a weak acetic acid solution. Addition of protease inhibitors while boiling resulted in somewhat lower yields, with one protein being totally absent with this method. Via mass spectrometry, the masses of one of these purified proteins was 13607.0Da (peak 2), which is consistent with the molecular weight of histone H2B. In addition, via western-blotting using anti-calf histone H2B antibody, all three proteins were positive and were thus named cvH2B-2, cvH2B-3 and cvH2B-4. The antibacterial activity of cvH2B-2 was similar to that of cvH2B-1, with activity against a Gram-positive bacterium (Lactococcus lactis subsp. lactis; minimum effective concentration [MEC] 52-57μg/mL) but inactive against Staphylococcus aureus (MEC>250μg/mL). However, both proteins had relatively potent activity against the Gram-negative oyster pathogen Vibrio parahemolyticus (MEC 11.5-14μg/mL) as well as the human pathogen Vibrio vulnificus (MEC 21.3-25.3μg/mL). cvH2B-3 and cvH2B-4 also had similarly strong activity against Vibrio vulnificus. These data provide further evidence for the antimicrobial function of histone H2B isomers in modulating bacterial populations in oyster tissues. The combined estimated concentrations of these histone H2B isomers were far above the inhibitory concentrations for the tested vibrios, including human pathogens. Our results indicate that the highly conserved histone proteins might be important components not only of immune defenses in oysters but have the potential to influence the abundance of a

  1. Torque modulates nucleosome stability and facilitates H2A/H2B dimer loss

    PubMed Central

    Sheinin, Maxim Y.; Li, Ming; Soltani, Mohammad; Luger, Karolin; Wang, Michelle D.

    2013-01-01

    The nucleosome, the fundamental packing unit of chromatin, has a distinct chirality: 147 bp of DNA are wrapped around the core histones in a left-handed, negative superhelix. It has been suggested that this chirality has functional significance, particularly in the context of the cellular processes that generate DNA supercoiling, such as transcription and replication. However, the impact of torsion on nucleosome structure and stability is largely unknown. Here we perform a detailed investigation of single nucleosome behavior on the high affinity 601 positioning sequence under tension and torque using the angular optical trapping technique. We find that torque has only a moderate effect on nucleosome unwrapping. In contrast, we observe a dramatic loss of H2A/H2B dimers upon nucleosome disruption under positive torque, while (H3/H4)2 tetramers are efficiently retained irrespective of torsion. These data indicate that torque could regulate histone exchange during transcription and replication. PMID:24113677

  2. Screening of lactic acid bacteria from vacuum packaged beef for antimicrobial activity

    PubMed Central

    Oliveira, Roseane B. P.; de L. Oliveira, Afonso; Glória, M. Beatriz A.

    2008-01-01

    The objective of this study was to isolate lactic acid bacteria (LAB) from vacuum packaged beef and to investigate their antagonist activity. LAB mean counts of 5.19 log cfu/cm2 were obtained from five samples of vacuum packaged beef. Two hundred isolates were selected and screened for the inhibitory effect on five ATCC reference Lactobacillus strains. Thirty six isolates showed activity in the agar spot test against at least two of the indicator strains. However, only six cell free supernatants (CFS) from these isolates exhibited activity against the indicator strains using the well-diffusion test and conditions that eliminated the effects of organic acids and hydrogen peroxide. L. acidophilus was the most sensitive indicator tested, whereas L. plantarum and L. fermentum were the most resistant ones. Identification by MIDI system indicated that these LAB isolates were Lactococcus lactis subsp. cremoris, Pediococcus acidilactici, Lactobacillus delbrueckii subsp. bulgaricus and Lactobacillus casei GC subgroup A. The antagonistic factors produced by most of these LAB against L. acidophilus were resistant to heat treatment (100°C for 10 min) and stable over a wide pH range (4.0 to 9.0). These data suggest that these isolates could be used as promising hurdles aiming increased safety and extended shelf life of meat products. PMID:24031232

  3. Pd-catalytic in situ generation of H2O2 from H2 and O2 produced by water electrolysis for the efficient electro-fenton degradation of rhodamine B.

    PubMed

    Yuan, Songhu; Fan, Ye; Zhang, Yucheng; Tong, Man; Liao, Peng

    2011-10-01

    A novel electro-Fenton process was developed for wastewater treatment using a modified divided electrolytic system in which H2O2 was generated in situ from electro-generated H2 and O2 in the presence of Pd/C catalyst. Appropriate pH conditions were obtained by the excessive H+ produced at the anode. The performance of the novel process was assessed by Rhodamine B (RhB) degradation in an aqueous solution. Experimental results showed that the accumulation of H2O2 occurred when the pH decreased and time elapsed. The maximum concentration of H2O2 reached 53.1 mg/L within 120 min at pH 2 and a current of 100 mA. Upon the formation of the Fenton reagent by the addition of Fe2+, RhB degraded completely within 30 min at pH 2 with a pseudo first order rate constant of 0.109 ± 0.009 min(-1). An insignificant decline in H2O2 generation and RhB degradation was found after six repetitions. RhB degradation was achieved by the chemisorption of H2O2 on the Pd/C surface, which subsequently decomposed into •OH upon catalysis by Pd0 and Fe2+. The catalytic decomposition of H2O2 to •OH by Fe2+ was more powerful than that by Pd0, which was responsible for the high efficiency of this novel electro-Fenton process.

  4. Fermented Acanthopanax koreanum Root Extract Reduces UVB- and H2O2-Induced Senescence in Human Skin Fibroblast Cells.

    PubMed

    Park, Min-Ja; Bae, Young-Seuk

    2016-07-28

    The present study assessed the effects of an aqueous extract of Acanthopanax koreanum root (AE) and of AE following fermentation by lactic acid bacteria (Lactobacillus plantarum and Bifidobacterium bifidum) (AEF) on human skin fibroblast HS68 cells exposed to ultraviolet B (UVB) irradiation and oxidative stress. AEF effectively antagonized the senescence-associated β-galactosidase staining and upregulation of p53 and p21(Cip1/WAF1) induced by UVB or H2O2 treatment in HS68 cells. It also exhibited excellent antioxidant activities in radical scavenging assays and reduced the intracellular level of reactive oxygen species induced by UVB or H2O2 treatment. The antioxidant and antisenescent activities of AEF were greater than those of nonfermented A. koreanum extract. AEF significantly repressed the UVB- or H2O2-induced activities of matrix metalloproteinase (MMP)-1 and -3, overexpression of MMP-1, and nuclear factor κB (NF-κB) activation. This repression of NF-κB activation and MMP-1 overexpression was attenuated by a mitogen-activated protein kinase activator, suggesting that this AEF activity was dependent on this signaling pathway. Taken together, these data indicated that AEF-mediated antioxidant and anti-photoaging activities may produce anti-wrinkle effects on human skin.

  5. Genome Sequence Analysis of the Biogenic Amine-Degrading Strain Lactobacillus casei 5b

    PubMed Central

    Ladero, Victor; Herrero-Fresno, Ana; Martinez, Noelia; del Río, Beatriz; Linares, Daniel M.; Fernández, María; Martín, María Cruz

    2014-01-01

    We here report a 3.02-Mbp annotated draft assembly of the Lactobacillus casei 5b genome. The sequence of this biogenic amine-degrading dairy isolate may help identify the mechanisms involved in the catabolism of biogenic amines and perhaps shed light on ways to reduce the presence of these toxic compounds in food. PMID:24435875

  6. Discovery of an acidic, thermostable and highly NADP+ dependent formate dehydrogenase from Lactobacillus buchneri NRRL B-30929

    USDA-ARS?s Scientific Manuscript database

    Objectives: To identify a robust NADP+ dependent formate dehydrogenase from Lactobacillus buchneri NRRL B-30929 (LbFDH) with unique biochemical properties. Results: A new NADP+ dependent formate dehydrogenase gene (fdh) was cloned from genomic DNA of L. buchneri NRRL B-30929. The recombinant constru...

  7. The in vitro antimicrobial activity of natural infant fluoride-free toothpastes on oral micro-organisms.

    PubMed

    Carvalho, Fabíola G; Negrini, Thais De Cássia; Sacramento, Luis Victor S; Hebling, Josimeri; Spolidorio, Denise M P; Duque, Cristiane

    2011-01-01

    The objective of this study was to evaluate the antimicrobial activity of six toothpastes for infants: 3 fluoride-free experimental toothpastes--cashew-based, mango-based and without plant extract and fluoride compared with 2 commercially fluoride-free toothpastes and 1 fluoridated toothpastes. Six toothpastes for infants were evaluated in this study: (1) experimental cashew-based toothpaste; (2) experimental mango-based toothpaste; (3) experimental toothpaste without plant extract and fluoride (negative control); (4) First Teeth brand toothpaste; (5) Weleda brand toothpaste; and (6) Tandy brand toothpaste (positive control). The antimicrobial activity was recorded against Streptococcus mutans, Streptococcus sobrinus, Lactobacillus acidophilus, and Candida albicans using the agar plate diffusion test. First Teeth, Weleda, mango-based toothpaste, and toothpaste without plant extract presented no antimicrobial effect against any of the tested micro-organisms. Cashew toothpaste had antimicrobial activity against S mutans, S sobrinus, and L acidophilus, but it showed no antimicrobial activity against C albicans. There was no statistical difference between the inhibition halo of cashew and Tandy toothpastes against S mutans and L acidophilus. Cashew fluoride-free toothpaste had inhibitory activity against Streptococcus mutans and Lactobacillus acidophilus, and these results were similar to those obtained for fluoridated toothpaste.

  8. Ca2+-Citrate Uptake and Metabolism in Lactobacillus casei ATCC 334

    PubMed Central

    Mortera, Pablo; Pudlik, Agata; Magni, Christian; Alarcón, Sergio

    2013-01-01

    The putative citrate metabolic pathway in Lactobacillus casei ATCC 334 consists of the transporter CitH, a proton symporter of the citrate-divalent metal ion family of transporters CitMHS, citrate lyase, and the membrane-bound oxaloacetate decarboxylase complex OAD-ABDH. Resting cells of Lactobacillus casei ATCC 334 metabolized citrate in complex with Ca2+ and not as free citrate or the Mg2+-citrate complex, thereby identifying Ca2+-citrate as the substrate of the transporter CitH. The pathway was induced in the presence of Ca2+ and citrate during growth and repressed by the presence of glucose and of galactose, most likely by a carbon catabolite repression mechanism. The end products of Ca2+-citrate metabolism by resting cells of Lb. casei were pyruvate, acetate, and acetoin, demonstrating the activity of the membrane-bound oxaloacetate decarboxylase complex OAD-ABDH. Following pyruvate, the pathway splits into two branches. One branch is the classical citrate fermentation pathway producing acetoin by α-acetolactate synthase and α-acetolactate decarboxylase. The other branch yields acetate, for which the route is still obscure. Ca2+-citrate metabolism in a modified MRS medium lacking a carbohydrate did not significantly affect the growth characteristics, and generation of metabolic energy in the form of proton motive force (PMF) was not observed in resting cells. In contrast, carbohydrate/Ca2+-citrate cometabolism resulted in a higher biomass yield in batch culture. However, also with these cells, no generation of PMF was associated with Ca2+-citrate metabolism. It is concluded that citrate metabolism in Lb. casei is beneficial when it counteracts acidification by carbohydrate metabolism in later growth stages. PMID:23709502

  9. Characterization of Cinnamoyl Esterases from Different Lactobacilli and Bifidobacteria.

    PubMed

    Fritsch, Caroline; Jänsch, André; Ehrmann, Matthias A; Toelstede, Simone; Vogel, Rudi F

    2017-02-01

    A high variety of plants that are used for food production contain esterified hydroxycinnamic acids. As their free forms display several benefits, like an enhanced absorption in human intestinal tract, anti-oxidative and anti-carcinogenic effects, an improved protein solubility and reduced discoloration, the microbial ability to cleave the ester bond is highly desired. In order to examine potential fermentation strains for this purpose, six different lactic acid bacteria and one bifidobacterial strain were screened for their ability to degrade esterified hydroxycinnamic acids because these strains are commonly used for fermentation of plant-based foods. Moreover, their cinnamoyl esterase activity was examined by molecular biological analyses. The enzymes were heterologously expressed in Escherichia coli, purified and biochemically characterized. The purified esterases with a molecular mass around 27-29 kDa had their optimum predominantly between pH 7 and 8 at 20-30 °C. Bifidobacterium animalis subsp. lactis, Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus plantarum and Lactobacillus fermentum displayed activities against a broad substrate range (methyl caffeate, methyl trans-p-coumarate, chlorogenic acid as well as partially ethyl ferulate). Concerning substrate affinity, reaction velocity, thermal and pH stability, Lactobacillus gasseri showed the overall best performance. The herein studied lactic acid- and bifidobacteria are promising for the production of fermented plant-based foods with an increased quality and nutritional value.

  10. CPsuperH2.0: An improved computational tool for Higgs phenomenology in the MSSM with explicit CP violation

    NASA Astrophysics Data System (ADS)

    Lee, J. S.; Carena, M.; Ellis, J.; Pilaftsis, A.; Wagner, C. E. M.

    2009-02-01

    We describe the Fortran code CPsuperH2.0, which contains several improvements and extensions of its predecessor CPsuperH. It implements improved calculations of the Higgs-boson pole masses, notably a full treatment of the 4×4 neutral Higgs propagator matrix including the Goldstone boson and a more complete treatment of threshold effects in self-energies and Yukawa couplings, improved treatments of two-body Higgs decays, some important three-body decays, and two-loop Higgs-mediated contributions to electric dipole moments. CPsuperH2.0 also implements an integrated treatment of several B-meson observables, including the branching ratios of B→μμ, B→ττ, B→τν, B→Xγ and the latter's CP-violating asymmetry A, and the supersymmetric contributions to the Bs,d0-B¯s,d0 mass differences. These additions make CPsuperH2.0 an attractive integrated tool for analyzing supersymmetric CP and flavour physics as well as searches for new physics at high-energy colliders such as the Tevatron, LHC and linear colliders. Program summaryProgram title: CPsuperH2.0 Catalogue identifier: ADSR_v2_0 Program summary URL:http://cpc.cs.qub.ac.uk/summaries/ADSR_v2_0.html Program obtainable from: CPC Program Library, Queen's University, Belfast, N. Ireland Licensing provisions: Standard CPC licence, http://cpc.cs.qub.ac.uk/licence/licence.html No. of lines in distributed program, including test data, etc.: 13 290 No. of bytes in distributed program, including test data, etc.: 89 540 Distribution format: tar.gz Programming language: Fortran 77 Computer: PC running under Linux and computers in Unix environment Operating system: Linux RAM: 32 Mbytes Classification: 11.1 Catalogue identifier of the previous version: ADSR_v1_0 Journal reference of the previous version: CPC 156 (2004) 283 Does the new version supersede the previous version?: Yes Nature of problem: The calculations of mass spectrum, decay widths and branching ratios of the neutral and charged Higgs bosons in the Minimal

  11. Lactobacillus pentosus B231 Isolated from a Portuguese PDO Cheese: Production and Partial Characterization of Its Bacteriocin.

    PubMed

    Guerreiro, Joana; Monteiro, Vitor; Ramos, Carla; Franco, Bernadette Dora Gombossy de Melo; Martinez, Rafael Chacon Ruiz; Todorov, Svetoslav Dimitrov; Fernandes, Paulo

    2014-06-01

    Bacteriocin B231 produced by Lactobacillus pentosus, isolated from an artisanal raw cow's milk protected designation of origin Portuguese cheese, is a small protein with an apparent relative mass of about 5 kDa and active against a large number of Listeria monocytogenes wild-type strains, Listeria ivanovii and Listeria innocua. Bacteriocin B231 production is highly dependent on the type of the culture media used for growth of Lact. pentosus B231. Replacement of glucose with maltose yielded the highest bacteriocin production from eight different carbon sources. Similar results were recorded in the presence of combination of glucose and maltose or galactose. Production of bacteriocin B231 reached maximal levels of 800 AU/ml during the stationary phase of growth of Lact. pentosus B231 in MRS broth at 30 °C. Bacteriocin B231 (in cell-free supernatant) was sensitive to treatment with trypsin and proteinase K, but not affected by the thermal treatment in range of 55-121 °C, or freezing (-20 °C). Bacteriocin production and inhibitory spectrum were evaluated. Gene encoding plantaricin S has been detected in the genomic DNA. Virulence potential and safety of Lact. pentosus B231 were assessed by PCR targeted the genes gelE, hyl, asa1, esp, cylA, efaA, ace, vanA, vanB, hdc1, hdc2, tdc and odc. The Lact. pentosus B231 strains harbored plantaricin S gene, while the occurrence of virulence, antibiotic resistance and biogenic amine genes was limited to cytolysin, hyaluronidase, aggregation substance, adhesion of collagen protein, gelatinase, tyrosine decarboxylase and vancomycin B genes.

  12. Phosphodiesterase inhibitors suppress Lactobacillus casei cell-wall-induced NF-κB and MAPK activations and cell proliferation through protein kinase A--or exchange protein activated by cAMP-dependent signal pathway.

    PubMed

    Saito, Takekatsu; Sugimoto, Naotoshi; Ohta, Kunio; Shimizu, Tohru; Ohtani, Kaori; Nakayama, Yuko; Nakamura, Taichi; Hitomi, Yashiaki; Nakamura, Hiroyuki; Koizumi, Shoichi; Yachie, Akihiro

    2012-01-01

    Specific strains of Lactobacillus have been found to be beneficial in treating some types of diarrhea and vaginosis. However, a high mortality rate results from underlying immunosuppressive conditions in patients with Lactobacillus casei bacteremia. Cyclic AMP (cAMP) is a small second messenger molecule that mediates signal transduction. The onset and progression of inflammatory responses are sensitive to changes in steady-state cAMP levels. L. casei cell wall extract (LCWE) develops arteritis in mice through Toll-like receptor-2 signaling. The purpose of this study was to investigate whether intracellular cAMP affects LCWE-induced pathological signaling. LCWE was shown to induce phosphorylation of the nuclear factor κB (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways and cell proliferation in mice fibroblast cells. Theophylline and phosphodiesterase inhibitor increased intracellular cAMP and inhibited LCWE-induced cell proliferation as well as phosphorylation of NF-κB and MAPK. Protein kinase A inhibitor H89 prevented cAMP-induced MAPK inhibition, but not cAMP-induced NF-κB inhibition. An exchange protein activated by cAMP (Epac) agonist inhibited NF-κB activation but not MAPK activation. These results indicate that an increase in intracellular cAMP prevents LCWE induction of pathological signaling pathways dependent on PKA and Epac signaling.

  13. Susceptibility of Oral Bacteria to an Antimicrobial Decapeptide

    DTIC Science & Technology

    2003-01-01

    coccus mitis ATCC 15913, Streptococcus oralis ATCC 35037T, Lactoba- cillus salivarius ATCC 29602, Lactobacillus acidophilus ATCC 43571 and...exhibited an ED99 (the dose at which 99% killing was observed after 15 min at 37 8C) of 6.25 gml1 against selected strains of Lactobacillus salivarius...broth microdilution assay. Growth of the cariogenic bacteria S.mutansATCC 25175T, S. sobrinus and L. acidophilus was also inhibited effectively by Fig

  14. 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

  15. In-Vitro Activity of Saponins of Bauhinia Purpurea, Madhuca Longifolia, Celastrus Paniculatus and Semecarpus Anacardium on Selected Oral Pathogens

    PubMed Central

    Jyothi, K. S.; Seshagiri, M.

    2012-01-01

    Objective: Dental caries, periodontitis and other mucosal diseases are caused by a complex community of microorganisms. This study aimed to investigate the antimicrobial properties of saponins of four important oil yielding medicinal plant extracts on selected oral pathogens that are involved in such diseases. Materials and Methods: Saponins were extracted from Bauhinia purpurea, Madhuca longifolia, Celastrus paniculatus and Semecarpus anacardium and purified. Antimicrobial properties of these saponins against Streptococcus mutans, Streptococcus mitis, Streptococcus salivarius, Staphylococcus aureus and Lactobacillus acidophilus were determined using well diffusion method. The minimum inhibitory concentration (MIC) was determined as the lowest concentration of saponins inhibiting bacterial growth after 14 h of incubation at 37°C. The bactericidal activity was evaluated using the viable cell count method. Results: The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Madhuca longifolia saponin on Streptococcus mutans MTCC 890, Streptococcus mitis and Staphylococcus aureus was 18.3 ± 0.15/34.4 ± 0.24 μg/ml, 19.0 ± 0.05/32.2 ± 0.0 μg/ml and 21.2 ± 0.35/39.0 ± 0.30 μg/ml, respectively and Bauhinia purpurea saponin on Streptococcus mutans MTCC 890, Staphylococcus aureus and Lactobacillus acidophilus was 26.4 ± 0.20/43.0 ± 0.40 μg/ml, 29.0 ± 0.30/39.6 ± 0.12 μg/ml and 20.2 ± 0.05/36.8 ± 0.23 μg/ml, respectively. Conclusion: The strong antimicrobial activity of Madhuca longifolia and Bauhinia purpurea may be due to the presence of complex triterpenoid saponins, oleanane type triterpenoid glycosides or atypical pentacyclic triterpenoid saponin. Hence, these extracted saponins may be used in food and oral products to prevent and control oral diseases. PMID:23323183

  16. Arginine Catabolism by Sourdough Lactic Acid Bacteria: Purification and Characterization of the Arginine Deiminase Pathway Enzymes from Lactobacillus sanfranciscensis CB1

    PubMed Central

    De Angelis, Maria; Mariotti, Liberato; Rossi, Jone; Servili, Maurizio; Fox, Patrick F.; Rollán, Graciela; Gobbetti, Marco

    2002-01-01

    The cytoplasmic extracts of 70 strains of the most frequently isolated sourdough lactic acid bacteria were screened initially for arginine deiminase (ADI), ornithine transcarbamoylase (OTC), and carbamate kinase (CK) activities, which comprise the ADI (or arginine dihydrolase) pathway. Only obligately heterofermentative strains such as Lactobacillus sanfranciscensis CB1; Lactobacillus brevis AM1, AM8, and 10A; Lactobacillus hilgardii 51B; and Lactobacillus fructivorans DD3 and DA106 showed all three enzyme activities. Lactobacillus plantarum B14 did not show CK activity. L. sanfranciscensis CB1 showed the highest activities, and the three enzymes were purified from this microorganism to homogeneity by several chromatographic steps. ADI, OTC, and CK had apparent molecular masses of ca. 46, 39, and 37 kDa, respectively, and the pIs were in the range of 5.07 to 5.2. The OTCs, CKs, and especially ADIs were well adapted to pH (acidic, pH 3.5 to 4.5) and temperature (30 to 37°C) conditions which are usually found during sourdough fermentation. Internal peptide sequences of the three enzymes had the highest level of homology with ADI, OTC, and CK of Lactobacillus sakei. L. sanfranciscensis CB1 expressed the ADI pathway either on MAM broth containing 17 mM arginine or during sourdough fermentation with 1 to 43 mM added arginine. Two-dimensional electrophoresis showed that ADI, OTC, and CK were induced by factors of ca. 10, 4, and 2 in the whole-cell extract of cells grown in MAM broth containing 17 mM arginine compared to cells cultivated without arginine. Arginine catabolism in L. sanfranciscensis CB1 depended on the presence of a carbon source and arginine; glucose at up to ca. 54 mM did not exert an inhibitory effect, and the pH was not relevant for induction. The pH of sourdoughs fermented by L. sanfranciscensis CB1 was dependent on the amount of arginine added to the dough. A low supply of arginine (6 mM) during sourdough fermentation by L. sanfranciscensis CB1

  17. The zinc finger gene Krox20 regulates HoxB2 (Hox2.8) during hindbrain segmentation.

    PubMed

    Sham, M H; Vesque, C; Nonchev, S; Marshall, H; Frain, M; Gupta, R D; Whiting, J; Wilkinson, D; Charnay, P; Krumlauf, R

    1993-01-29

    The zinc finger gene Krox20 and many Hox homeobox genes are expressed in segment-restricted domains in the hindbrain. The restricted expression patterns appear before morphological segmentation, suggesting that these transcription factors may play an early role in the establishment and identity of rhombomeric segments. In this paper, we show that the HoxB2 (Hox2.8) gene is normally upregulated in rhombomeres (r) 3, 4, and 5, and we identify an enhancer region upstream of the gene that imposes r3/r5 expression in transgenic mice. This enhancer contains three Krox20-binding sites required in vitro for complex formation with Krox20 protein and in vivo for rhombomere-restricted expression. In transgenic mice, Krox20 expressed in ectopic domains can transactivate a reporter construct containing the HoxB2 r3/r5 enhancer. These data demonstrate that Krox20 is a part of the upstream transcriptional cascade that directly regulates HoxB2 expression during hindbrain segmentation.

  18. Bovine mastitis may be associated with the deprivation of gut Lactobacillus.

    PubMed

    Ma, C; Zhao, J; Xi, X; Ding, J; Wang, H; Zhang, H; Kwok, L Y

    2016-02-01

    Bovine mastitis is an economical important microbial disease in dairy industry. Some recent human clinical trials have shown that oral probiotics supplementation could effectively control clinical mastitis, suggesting that the mechanism of mastitis protection might be achieved via the host gut microbiota. We aimed to test our hypothesis that bovine mastitis was related to changes in both the mammary and gut microbial profiles. By quantitative PCR, the milk and faecal microbial profiles of cows with low (<3×10 5 cells/ml) and high (>1×10 6 cells/ml) somatic cell count (SCC) were compared. Firstly, we observed drastic differences in both the milk and faecal microbial compositions at genus and Lactobacillus-species levels between the two groups. Secondly, the pattern of faecal microbial community changes of mastitis cows was similar to that of the milk, characterised by a general increase in the mastitis pathogens (Enterococcus, Streptococcus and Staphylococcus) and deprivation of Lactobacillus and its members (L. salivarius, L. sakei, L. ruminis, L. delbrueckii, L. buchneri, and L. acidophilus). Thirdly, only the faecal lactobacilli, but not bifidobacteria correlated with the milk microbial communities and SCC. Our data together hint to a close association between bovine mastitis, the host gut and milk microbiota.

  19. Modeling Lactic Fermentation of Gowé Using Lactobacillus Starter Culture

    PubMed Central

    de J. C. Munanga, Bettencourt; Loiseau, Gérard; Grabulos, Joël; Mestres, Christian

    2016-01-01

    A global model of the lactic fermentation step of gowé was developed by assembling blocks hosting models for bacterial growth, lactic acid production, and the drop of pH during fermentation. Commercial strains of Lactobacillus brevis and of Lactobacillus plantarum were used; their growth was modeled using Rosso’s primary model and the gamma concept as a secondary model. The optimum values of pH and temperature were 8.3 ± 0.3, 44.6 ± 1.2 °C and 8.3 ± 0.3, 3.2 ± 37.1 °C with μmax values of 1.8 ± 0.2 and 1.4 ± 0.1 for L. brevis and L. plantarum respectively. The minimum inhibitory concentration of undissociated lactic acid was 23.7 mM and 35.6 mM for L. brevis and L. plantarum, respectively. The yield of lactic acid was five times higher for L. plantarum than for L. brevis, with a yield of glucose conversion to lactic acid close to 2.0 for the former and 0.8 for the latter. A model was developed to predict the pH drop during gowé fermentation. The global model was partially validated during manufacturing of gowé. The global model could be a tool to aid in the choice of suitable starters and to determine the conditions for the use of the starter. PMID:27916901

  20. Testing of viscous anti-HIV microbicides using Lactobacillus.

    PubMed

    Moncla, B J; Pryke, K; Rohan, L C; Yang, H

    2012-02-01

    The development of topical microbicides for intravaginal use to prevent HIV infection requires that the drugs and formulated products be nontoxic to the endogenous vaginal Lactobacillus. In 30min exposure tests we found dapivirine, tenofovir and UC781 (reverse transcriptase inhibitor anti-HIV drugs) as pure drugs or formulated as film or gel products were not deleterious to Lactobacillus species; however, PSC-RANTES (a synthetic CCR5 antagonist) killed 2 strains of Lactobacillus jensenii. To demonstrate the toxicity of formulated products a new assay was developed for use with viscous and non-viscous samples that we have termed the Lactobacillus toxicity test. We found that the vortex mixing of vaginal Lactobacillus species can lead to reductions in bacterial viability. Lactobacillus can survive briefly, about 2s, but viability declines with increased vortex mixing. The addition of heat inactivated serum or bovine serum albumin, but not glycerol, prevented the decrease in bacterial viability. Bacillus atrophaeus spores also demonstrated loss of viability upon extended mixing. We observed that many of the excipients used in film formulation and the films themselves also afford protection from the killing during vortex mixing. This method is of relevance for toxicity for cidal activities of viscous products. Copyright © 2011 Elsevier B.V. All rights reserved.