Evaluation of probiotic properties of Lactobacillus plantarum strains isolated from Chinese sauerkraut

Lactobacillus plantarum strains isolated and identified from naturally-fermented Chinese sauerkraut were examined in vitro for potential probiotic properties and in vivo for cholesterol-lowering effect in mice. Among 7 isolated L. plantarum strains, strains S2-5 and S4-1 were found to possess desirable probiotic properties including ability to survive at pH 2.0 for 60 min, tolerate pancreatin and bile salts, adhere to Caco-2 cells, produce high β-galactosidase activity and antimicrobial activity against Escherichia coli O157 and Shigella flexneri CMCC(B). In addition, strains S2-5 and S4-1 were susceptible to several antibiotics, and capable of reducing cholesterol level in MRS medium by assimilation of cholesterol at 20.39 and 22.28 μg ml^?1, respectively. The in vivo study with L. plantarum S4-1 showed that feeding with fermented milk containing this strain was able to effectively reduce serum cholesterol level in mice, demonstrating its potential as an excellent probiotic candidate for applications in functional products.     

Determining potential probiotic properties of human originated Lactobacillus plantarum strains

In this study, twenty Lactobacillus plantarum strains which were isolated from the fecal samples of humans were investigated in vitro for their characteristics as potential new probiotic strains. The L. plantarum strains were examined for resistance to gastric acidity in simulated gastric juice at pH 2.0, 2.5, 3.0, and 3.5. The growth of test cultures with different pH was monitored after 0, 10, 30, 60, 90, and 120 min of incubation using a spectrophotometer at 550 nm. At the same time, samples were serially diluted in sterile PBS, and counts of viable bacteria were determined by plate counts using MRS agar for each pH and time parameter. The strains were also examined for resistance to 0.4% phenol, production of H₂O₂, adhesion to Caco-2 cell line and antimicrobial activity. It was determined that the artificial gastric juice, even at pH 2.0, did not significantly change the viability of the cultures. Except L. plantarum AA1-2, all strains were detected at 8 ～ 9 log₁₀ CFU/g. It was found that all L. plantarum strains showed good resistance to 0.4% phenol, and only one strain (AC18-82) produced H₂O₂. Good adhesion of L. plantarum strains to Caco-2 cells was observed in this experiment. These selected strains also showed antimicrobial activity.     

Antifungal activity of two Lactobacillus strains with potential probiotic properties

Aflatoxin (highly toxic and carcinogenic secondary metabolites produced by fungi) contamination is a serious problem worldwide. Modern agriculture and animal production systems need to use high-quality and mycotoxin-free feedstuffs. The use of microorganisms to preserve food has gained importance in recent years due to the demand for reduced use of chemical preservatives by consumers. Lactic acid bacteria are known to produce various antimicrobial compounds that are considered to be important in the biopreservation of food and feed. Lactobacillus rhamnosus L60 and Lactobacillus fermentum L23 are producers of secondary metabolites, such as organic acids, bacteriocins and, in the case of L60, hydrogen peroxide. The antifungal activity of lactobacilli strains was determined by coculture with Aspergillus section Flavi strains by two qualitative and one quantitative methods. Both L23 and L60 completely inhibited the fungal growth of all aflatoxicogenic strains assayed. Aflatoxin B (1) production was reduced 95.7-99.8% with L60 and 27.5-100% with L23. Statistical analysis of the data revealed the influence of L60 and L23 on growth parameters and aflatoxin B (1) production. These results are important given that these aflatoxicogenic fungi are natural contaminants of feed used for animal production, and could be effectively controlled by Lactobacillus L60 and L23 strains with probiotic properties.     

Different probiotic properties for Lactobacillus fermentum strains isolated from swine and poultry

Systematic procedures were used to evaluate the probiotic properties of Lactobacillus fermentum (L. fermentum) strains isolated from swine and poultry. The major properties included their capabilities to adhere to the intestinal epithelium of swine and poultry, the inhibition on pathogenic bacteria, and their tolerance to the gastric juice and bile salts. Results showed that L. fermentum strains from poultry digestive tract showed better adherence to the swine intestine and chicken crop epithelial cells as compared to those strains from the swine origin. In addition, six strains from poultry and one strain from swine showed adhesion specificity to their own intestinal epithelium. Four poultry isolates and one swine isolate were able to adhere to the epithelial cells from both swine and chicken. For gastric juice and bile tolerance, most of the strains isolated from swine or poultry were acid tolerant but less strains were bile intolerant. The spent culture supernatant (SCS) of these L. fermentum strains showed antagonistic effect against the indicator bacteria, such as Escherichia coli, Salmonella spp., Shigella sonnei and some enterotoxigenic Staphylococcus aureus. From the above studies, some L. fermentum strains isolated from poultry were found to have the probiotic properties required for use in animal feed supplement. This study suggested that poultry digestive tract may serve as potential source for the isolation of probiotic lactic acid bacteria.     

Assessment of cell surface properties and adhesion potential of selected probiotic strains

Aim:  To evaluate the physicochemical cell surface and adhesive properties of selected probiotic strains for human use.
Methods and Results:  Probiotic strains, Bifidobacterium longum B6, Lactobacillus acidophilus ADH, Lactobacillus paracasei , Lactobacillus rhamnosus GG, Lactobacillus brevis , Lactobacillus casei , Leuconostoc mesenteroides and Pediococcus acidilactici were tested for the physicochemical properties of cell surfaces and the adhesion abilities against foodborne pathogens. Bif .  longum B6 (53·6%) and Lact .  rhamnosus GG (46·5%) showed the highest hydrophobicity, while the least affinity to xylene was observed in Ped .  acidilactici (10·4%). Bifidobacterium longum B6 showed the strongest coaggregation phenotype with Listeria monocytogenes (53·0%), Shigella boydii (42·0%) and Staphylococcus aureus (45·9%). Lactobacillus rhamnosus GG had the strong binding ability to Caco-2 cells and effectively inhibited the adhesion of L .  monocytogenes , Salmonella Typhimurium, Sh .  boydii and Staph .  aureus to Caco-2 cells. The hydrophobicity was highly correlated with coaggregative abilities and competitive inhibition, suggesting a good relationship between in vitro adhesion and in vivo colonization.
Conclusion:  The results suggest that Bif .  longum B6 and Lact .  rhamnosus GG can be candidate probiotics available for human consumption.
Significance and Impact of the Study:  Because the use of probiotic strains has been more concerned with their beneficial effects in the GI tract, it is essential to examine the potential of probiotic strains based on the physicochemical properties in terms of bacterial-binding and adhesion capabilities.     

Metabolism of biodiesel-derived glycerol in probiotic Lactobacillus strains

Three probiotic Lactobacillus strains, Lactobacillus acidophilus, Lactobacillus plantarum, and Lactobacillus delbrueckii, were tested for their ability to assimilate and metabolize glycerol. Biodiesel-derived glycerol was used as the main carbon and energy source in batch microaerobic growth. Here, we show that the tested strains were able to assimilate glycerol, consuming between 38 and 48 % in approximately 24 h. L. acidophilus and L. delbrueckii showed a similar growth, higher than L. plantarum. The highest biomass reached was 2.11 g?L^?1 for L. acidophilus, with a cell mass yield (Y _X/S) of 0.37 g?g^?1. L. delbrueckii and L. plantarum reached a biomass of 2.06 and 1.36 g?L^?1. All strains catabolize glycerol mainly through glycerol kinase (EC 2.7.1.30). For these lactobacillus species, kinetic parameters for glycerol kinase showed Michaelis–Menten constant (K _m) ranging from 1.2 to 3.8 mM. The specific activities for glycerol kinase in these strains were in the range of 0.18 to 0.58 U?mg?protein^?1, with L. acidophilus ATCC 4356 showing the maximum specific activity after 24 h of cultivation. Glycerol dehydrogenase activity was also detected in all strains studied but only for the reduction of glyceraldehyde with NADPH (K _m for DL-glyceraldehyde ranging from 12.8 to 32.3 mM). This enzyme shows a very low oxidative activity with glycerol and NADP⁺ and, most likely, under physiological conditions, the oxidative reaction does not occur, supporting the assumption that the main metabolic flux concerning glycerol metabolism is through the glycerol kinase pathway.     

Selenite-stress selected mutant strains of probiotic bacteria for Se source production

Selenium deficiency is a major health problem worldwide for about 1 billion people. Bacterial cells usually possess low tolerance to selenite stress and also low ability to reduce high concentrations of toxic selenite. Here, high tolerance to selenite and selenium bioaccumulation capability were developed in mutated clones of probiotic and starter bacteria including Enterococcus faecium, Bifidobacterium animalis ssp. lactis, Lactobacillus casei and Lactococcus lactis ssp. lactis by food-level strain development process and clone selection. All mutant clones possessed increased glutathione concentration and glutathione reductase activity. The selenite treatment increased further these values in L. casei mutant strain pointing at a different selenite reduction pathway and/or stress response in this organism. Considerable conversion of selenite to cell bound selenium forms with a concomitant high biomass production was detected in E. faecium and B. animalis ssp. lactis cultures. Possible application of these strains as food and feed supplements is under investigation.     

从实际应用特点出发筛选益生乳杆菌菌株

正众所周知,在人的体内和体表生活着大量的各类共生微生物。后者的总数量可达人体自身细胞数量的10-100倍,编码的基因数量可达人体自身的300多倍,被称为人体的"第二基因组",对保障人体的健康具[1]有重要意义。研发各种益生菌制剂来维持、调整、完善人体,特别是肠道的正常菌群结构,近年来一直[2]是备受关注的热点和前沿。乳杆菌作为公认的益生菌,可有效改善宿主的肠道环境、抑制有害菌的生长、     

The probiotic properties of Lactobacillus buchneri P2

Aims: To isolate new lactobacilli strain with cholesterol‐lowering effect and analyse its probiotic properties and possible mechanisms of cholesterol removal. Methods and Results: The strain with cholesterol‐lowering effect was isolated from pickled juice. The acid and bile tolerance and antimicrobial activity were tested. The free cholalic acid, the cholesterol in supernatant fluid, washing buffer and cell extract, the cholesterol removed by growing, dead and resting cells were quantified. The isolated strain with high cholesterol‐reducing rate of 43·95% was identified as Lactobacillus buchneri (Lact. buchneri) P2. It had acid and bile tolerance and antimicrobial activity. Moreover, it could remove cholesterol via coprecipitating with deconjugated bile salts, assimilating and adsorbing by cells. And the assimilation was considered to be the main reason of cholesterol removal. Conclusions: The isolated Lact. buchneri P2 showed probiotic properties of cholesterol reduction, acid and bile tolerance and antimicrobial activity and could remove cholesterol via different ways. Significance and Impact of the Study: A new strain of Lact. buchneri P2 with efficient cholesterol‐reducing ability was isolated to provide species diversity of lactobacilli for functional dairy products. And the possible mechanism of cholesterol removal by Lact. buchneri was discussed.     

Characterization of the three selected probiotic strains for the application in food industry

Previously selected bacterial probiotic strains Enterococcus faecium L3, Lactobacillus plantarum L4 and Lactobacillus acidophilus M92 have shown their potential as functional starter cultures in silage, white cabbage and milk fermentation. Therefore, the phenotypic and genotypic characteristics important for their application in food industry were investigated. Pulsed-field gel electrophoresis (PFGE) of NotI digested genomic DNA, in combination with physiological traits determined by API tests, made a useful tool for identification of these probiotic strains and differentiation among them. Lyophilized probiotic cells remained viable during 75 days of storage at −20, +4 and +15°C, while fresh concentrated cells remained viable only at −20°C with addition of glycerol as cryoprotectant. After the lyophilization with addition of skim milk as lyoprotectant, the viability of L. acidophilus M92, L. plantarum L4 and E. faecium L3 was reduced by only 0.37, 0.44 and 0.50 log, respectively. Furthermore, probiotic strains L. acidophilus M92, L. plantarum L4, and E. faecium L3, demonstrated anti-Salmonella activity, and L. acidophilus M92 having also antilisterial activity demonstrated by in vitro competition test. Overnight cultures and cell-free supernatants of the three probiotic strains exerted also an antagonistic effect against the Gram-positive and Gram-negative test microorganisms examined, demonstrated by the agar-well diffusion test. The inhibition of Listeria monocytogenes, Salmonella typhimurium, Yersinia enterocolitica, and Acinetobacter calcoaceticus obtained, achieved by the neutralized, 5-fold concentrated supernatant of L. plantarum L4, may be the result of its bacteriocinogenic activity. On the basis of these results, the application of the three examined probiotic strains may become a point of great importance in respect of food safety.     

Potential probiotic properties of phytase-producing Lactobacillus salivarius FC113

Probiotics have known efficacy as dietary supplements. Here, Lactobacillus strain F113 was characterized for probiotic use. Strain FC113 was selected as having the highest phytase activity (403.6 U) among tested strains showing acid tolerance and nitrite production. FC113 was tentatively identified as Lactobacillus salivarius based on an API 50 CHL assay and 16S rRNA gene analysis. The production of interleukin (IL)-1α and tumor necrosis factor (TNF)-α was measured in in vitro culture experiments. Cytokine production by L. salivarius FC113 at 1?×?10⁷ CFU/ml was approximately 175.5?±?36.40 pg/mL IL-1α and 353.5?±?61.79 pg/mL TNF-α. L. salivarius FC113 was profoundly resistant to artificial gastric juice (pH 2.5, 1 % pepsin), and persisted for 24 h in artificial bile acid. According to results obtained with an API ZYM kit, L. salivarius FC113 did not generate carcinogenic enzymes. L. salivarius F113 had an inhibitory effect on food-borne pathogens, and adhered strongly to HT-29 human intestinal epithelial cell lines. These results show that L. salivarius FC113 has probiotic characteristics, and exhibits high feed bioavailability in the host animal, in addition to an immune-stimulating effect.     

Revised classification of native probiotic strains of Lactobacillus used in Russian Federation

Thirteen strains of industrial bacterial cultures of the genus Lactobacillus (from a collection of Gabrichevsky Research Institute of Epidemiology and Microbiology) were studied. These strains were used for decades in Russian Federation for food and drug production, as ferments for lactic acid products, for production of probiotics, biologically active and veterinary preparations. Complex analysis of data on cultures obtained using microbiological and molecular-genetic methods was conducted for the first time. Biochemical characteristics of these cultures were studied and the sequence of the proximal region of 16S ribosomal RNA gene was determined. The employment of the test system API-50CHL was shown to broaden the opportunities of a more accurate biochemical identification of bacteria belonging to the genus Lactobacillus, in comparison with the set ANAEROTEST-23. According to the results obtained in a comparative analysis of nucleotide sequences of 16S rRNA gene, all strains examined show 97-99% homology of the proximal region of this gene with that of the type representatives of studied species. These data allowed taxonomic reclassification of the species position of cultures with consideration of the more advanced level of systematics. Nucleotide sequences of gene fragments of examined lactobacilli strains were recorded in NCBI database (accession numbers of deposits GU560031, GU560032, GU560033, GU560034, GU560035, GU560036, GU560037, GU560038, GU560039, GU560040, GU560041, GU560042, GU560043).     

Fermentation of coconut water by probiotic strains Lactobacillus acidophilus L10 and Lactobacillus casei L26

Coconut water is becoming an increasingly popular beverage and sports drink in tropical countries due to its high mineral content. Probiotic fermentation of coconut water would provide consumers with a novel probiotic beverage which can provide both hydration and probiotic benefits. The aim of this study was to assess the growth, survival and fermentation performance of two probiotic bacteria in coconut water. Lactobacillus acidophilus L10 and L. casei L26 grew well in coconut water and showed similar growth patterns. The viable cell count of the two probiotic cultures reached approximately 10⁸ CFU/ml after 2 days fermentation at 37 °C and maintained approximately10⁷–10⁸ CFU/ml after 26 days at 4 °C. Changes in total soluble solids (^oBrix), pH, sugars, organic acids and minerals were similar between the two probiotic cultures, except for fructose, glucose, copper, phosphorus and lactic, acetic and malic acids. There were significant variations between the two cultures in their ability to produce and consume these compounds. L. acidophilus produced higher amounts of 2-heptanone, 2-nonanone, benzaldehyde, 2-heptanol, 2-nonanol, δ-octalactone and δ-dodecalactone, whereas L. casei produced higher amounts of acetic acid, diacetyl, acetoin, δ-decalactone, 3-methyl-3-buten-1-ol, linalool, 1-octanol, p-tolualdehyde and ethyl 2-hydroxypropanoate. There was no substantial change in mineral content. These results suggest the feasibility of fermenting coconut water into a probiotic beverage, especially for sports nutrition, with the dual benefits of electrolytes and probiotics.     

罗伊乳杆菌LE16益生特性的研究

目的研究罗伊乳杆菌LE16的益生特性。方法通过体外和动物实验,评价罗伊乳杆菌LE16的益生特性。结果该菌株对p H 2.5的强酸和0.3%高胆盐环境均有良好的耐受性。对引起肠道感染的几种病原菌均有不同程度的拮抗,尤其对枯草芽胞杆菌、伤寒沙门菌、单核细胞增生李斯特菌及蜡样芽胞杆菌的抑制作用最强。对肠道菌群正常和失调的小鼠均具有调节功能。结论罗伊乳杆菌LE16具有良好的益生性能,可作为益生菌菌株开发应用于食品和保健食品中。     

Distinct immunomodulatory properties of Lactobacillus paracasei strains

Aims: This study was performed to ascertain the immunomodulatory effect of Lactobacillus paracasei strains. These strains were also genetically characterized. Methods and Results: The strains were genetically differentiated by using the fluorescent‐amplified fragment length polymorphism technique, which led to the identification of several molecular markers unique to each strain. To determine the immunomodulatory properties, we evaluated the effect of strains on dendritic cell maturation, dextran uptake, ability to induce proliferation of allogenic T cells and cytokine secretion. The results indicated that all the strains stimulated phenotypic maturation of dendritic cells (DCs), but they acted differently on DCs in relation to the other tested properties; notably, a different effect on cytokine secretion was detected. Conclusions: The results of this study revealed different immunomodulatory properties of strains of the species Lact. paracasei. Strain IMPC 4.1 showed an interesting anti‐inflammatory ability. Probiotic strains IMPC 2.1 and LMG P‐17806 were characterized by a similar and intermediate ability to induce cytokine secretion in contrast to the very low ability of strain LMG 23554. Significance and Impact of Study: Our results confirm that each single strain of a bacterial species appears to influence the immune system in a peculiar manner. The evaluation of the different types and/or levels of cytokines whose secretion is induced by each strain could be relevant to define its pro‐ or anti‐inflammatory properties and its more appropriate clinical use.