Probiotic Lactobacillus Acidophilus NCFM Research Articles

Bee pollen as a novel substrate in pilot-scale probiotic-mediated lactic fermentation processes

Different process conditions were evaluated to induce lactic acid fermentation of bee pollen at the pilot scale using three different commercial starter cultures. Clean and dry Colombian high Andean bee pollen was used as the sole source of nutrients (substrate). The effect of substrate preparation conditions (water:pollen ratio and initial substrate acidity) on lactic fermentation was evaluated by monitoring pH, titratable acidity, and pollen morphology in 6-kg-fermentation units. A 2:1 water:pollen mixture with an initial pH adjusted to 5.8, pretreated with heat/pressure (10 min; 80 kPa; 115 °C), was determined to be an adequate substrate for pilot-scale lactic fermentation for all lactic starter cultures. At the selected conditions, the kinetics of lactic acid production and the growth of specific lactic bacteria were determined. The fermentation process time was 30 h, after which a significant decline in bacterial populations was observed. Among the starter cultures tested, one constituted exclusively by probiotic Lactobacillus acidophilus NCFM presented the best performance in terms of lactic acid production, with a concentration of 1.65% wt, after 30 h. The results highlight the feasibility of producing fermented bee pollen at a commercial scale as a functional food product with probiotic features.

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

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. Biological significanceHuman diet is important for intestinal health and food components, especially non-digestible carbohydrates can beneficially modify the microbiota. In the present study, effects of emerging and established prebiotic carbohydrates on the probiotic potential of Lactobacillus acidophilus NCFM were investigated by testing adhesion to a mucin layer and intestinal cells, and comparing this with changes in abundancy of surface proteins thought to be important for host interactions. Increased adhesion was observed following culturing of the bacterium with fructooligosaccharides, cellobiose or polydextrose, as well as mucin-supplemented glucose as carbon source. Enhanced adhesion ability can prolong bacterial residence in GIT yielding positive health effects. Higher relative abundance of certain surface proteins under various conditions (i.e. grown on cellobiose or mucin-supplemented glucose) suggested involvement of these proteins in adhesion, as confirmed by competition in case of two recombinantly produced moonlighting proteins. Combination of Lactobacillus acidophilus NCFM with different carbohydrates revealed potential bacterial determinants of synbiotic interactions, including stimulation of adhesion.

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

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.

Cereal β-glucan immune modulating activity depends on the polymer fine structure

ScopeThe relationship between the immune modulatory properties of different mixed linkage β-glucans from plant and microbial origin and their physico-chemical properties is elusive. By the use of murine dendritic cells and multivariate data analysis, this study examines the importance of the physico-chemical characteristics of β-glucans for their capacity to modulate immune responses. Methods and resultsA total of 23 different β-glucan samples characterised with respect to compositional, molecular, structural and rheological properties were investigated for their immune modulatory properties. Most of the β-glucans showed a decrease in the LPS induced IL-12 production and an increase in the LPS induced IL-10 levels in dendritic cells. When the dendritic cells were stimulated with the probiotic Lactobacillus acidophilus NCFM, the β-glucans up-regulate the IL-12 production but showed no effect on the IL-10 levels. The modulation of the cytokine responses was shown to depend on the β-glucan block structure as expressed by the ratio of cellotriosyl to cellotetraosyl units in the molecule as well as the polymer solubility and aggregation in solution. ConclusionThis comparative study reveals that the β-glucan fine structure and in turn the intermolecular organisation in solution are key factors that determine their immune modulatory capacity.

Comparative fermentation of insoluble carbohydrates in an in vitro human feces model spiked with Lactobacillus acidophilus NCFM

AbstractDietary fiber and slow carbohydrates can have differential beneficial effects on gut microbial composition and metabolism. Insoluble substrates such as RS cannot be used in continuous simulation systems and we tested an in vitro batch fermentation system for possible prebiotic effects using waxy maize starch granules, pectin‐rich potato fiber, and potato lintner starch tested with human fresh fecal microbiota spiked with the probiotic Lactobacillus acidophilus NCFM. Microbial quantification by real‐time polymerase chain‐reaction (qRT‐PCR) revealed that Bacteroidetes was specifically suppressed by each insoluble carbohydrate resulting in a clear decrease in the ratio of Bacteroidetes and Firmicutes. Notably, all carbohydrates tested appeared to block the formation of the potentially harmful branched chain fatty acids (BCFA) fermentation products, but supported lactobacilli growth and increase in lactic acid production. Potato lintner starch had the greatest effect. Insoluble carbohydrates also suppressed production of SCFAs as compared to the control medium. Importantly, potato lintner starch most efficiently suppressed the ratio between the Bacteroidetes and Firmicutes and suppressed growth of bifidobacteria, Enterobactericeae and Faecalibacterium prausnizii. Degradative resistance of the lintner starch were linked to these effects. Problems using continuous fermentation systems for insoluble prebiotics and interpretation of batch fermentation data are discussed.

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

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 for pizza cheeses.

Gut microbiota of healthy elderly NSAID users is selectively modified with the administration of Lactobacillus acidophilus NCFM and lactitol

Ageing changes gut microbiota composition and alters immune system function. Probiotics, prebiotics and synbiotics may improve the health status of elderly individuals by modifying the intestinal environment and the microbiota composition, and by stimulating the immune system. In this work, we studied the effects of synbiotic supplementation on the gut microbiota of healthy elderly volunteers. Fifty-one elders were randomly assigned to consume either a synbiotic dietary supplement or a placebo in addition to their usual diet for a 2-week period. The synbiotic product consisted of the probiotic Lactobacillus acidophilus NCFM and the prebiotic lactitol and was ingested twice a day, with a total daily dose of 10g lactitol and 2 × 10(10) cells of probiotic bacteria. Before, during and after the intervention period fecal quantities of six phylogenetic bacterial groups were determined using quantitative PCR, and relative changes in total microbiota composition were assessed by percent guanine-plus-cytosine profiling. The microbiota profiles showed certain relative changes within the microbial community, and indicated an increase of bifidobacteria levels during synbiotic supplementation. Quantification by PCR confirmed the in changes in the microbiota composition; for example increases in total levels of endogenous bifidobacteria and lactobacilli were recorded. Throughout the 6-week study period there was a decrease unrelated to intervention in the Blautia coccoides-Eubacterium rectale bacterial group levels and Clostridium cluster XIVab levels, but this decrease appeared to be halted during the synbiotic intervention. In conclusion, putatively beneficial changes in microbiota were observed in the elderly subjects supplemented with the synbiotic product.

Prophylactic Feeding of Lactobacillus acidophilus NCFM to Mice Attenuates Overt Colonic Hyperplasia

The objective of this project was to determine if the probiotic Lactobacillus acidophilus NCFM would protect mice from developing transmissible murine colonic hyperplasia (TMCH) caused by Citrobacter rodentium. Our hypothesis was that the oral administration of L. acidophilus NCFM to mice would mitigate colonic hyperplasia and modulate the host immune response. A concurrent administration (CA) study was performed by feeding mice phosphate-buffered saline (PBS), C. rodentium only, L. acidophilus NCFM only, or C. rodentium and NCFM concurrently on the same day. The mice in the CA study were not protected by the probiotic, since their mean colon sample weights (0.109 g) were significantly higher than those of the PBS controls (0.0774 g), and the hematoxylin and eosin–stained samples showed histological changes typically associated with TMCH. A prophylactic feeding (PF) study was performed by orally feeding mice PBS or NCFM once daily for 20 consecutive days; in addition, on day 7, mice were challenged with either PBS or C. rodentium. Mice in the PF study were protected when they consumed the probiotic prior to the pathogen challenge, since their mean colon sample weights (0.0812 g) were not significantly higher than those of the controls (0.0753 g). The hematoxylin and eosin–stained samples appeared similar to the control samples, and the intestinal interleukin (IL)-15 and gamma interferon (IFN-γ) mRNA levels were reduced. L. acidophilus NCFM did attenuate overt colonic hyperplasia when fed to mice prior to challenge with C. rodentium. The mouse model used in this study enabled us to investigate the efficacy of the L. acidophilus NCFM in preventing gastrointestinal disease and is a valid model for future probiotic research.