We isolated several strains of human and animal origin, focusing on the genera Lactobacillus and Bifidobacterium. Samples of cow colostrum, feces and saliva of calves and piglets, and infant feces were acquired, bacterial strains were isolated and identified, and twenty strains of lactobacilli and bifidobacteria were shortlisted for further in vitro analysis of adhesion capacity to human cells, surface hydrophobicity, and auto-aggregation. Adhesion capacity was evaluated using a mixture of Caco-2 and HT-29-MTX cells and hydrophobicity was measured using the microbial adhesion to the hydrocarbon method. Lactobacillus reuteri was the most frequently isolated species, followed by L. casei subsp. paracasei and L. gasseri; all strains were isolated from infant feces. Bifidobacteria were represented by the species B. longum (infant feces), and B. thermophilum and B. pseudolongum (calves, piglets). All twenty strains showed good adhesion capacities to the mixed cell culture (17.7–37.2%), particularly, L. reuteri isolates K7 and K14 (37.2% and 35.5%, respectively). In hydrophobicity and auto-aggregation assays, strain-specific differences irrespective of the origin or taxonomic group were observed. Hydrophobicity values varied considerably (from 6.1% to 87.4%), whereas auto-aggregation ability ranged from 21.7% to 69.7%. No relation was observed between hydrophobicity and adhesion capacity; instead, auto-aggregation was apparently related with adhesion.
Pfaffia glomerata (Amaranthaceae) is popularly known as "Brazilian ginseng." Previous studies have shown that fructose is the major carbohydrate component present in its roots. Inulin-type fructans, polymers of fructose, are the most widespread and researched prebiotics. Here, we isolated and chemically characterized inulin extracted from P. glomerata roots and investigated its potential prebiotic effect. Fructans were isolated and their structures were determined using colorimetric, chromatography, polarimetry, and spectroscopic analysis. The degree of polymerization (DP) was determined, and an in vitro prebiotic test was performed. The structure of inulin was confirmed by chromatography and spectroscopic analysis and through comparison with existing data. Representatives from the genera Lactobacillus and Bifidobacterium utilized inulin from P. glomerata, because growth was significantly stimulated, while this ability is strain specific. The results indicated that inulin extracted from P. glomerata roots represents a promising new source of inulin-type prebiotics.
Stevia rebaudiana (Bertoni) is widely studied because of its foliar steviol glycosides. Fructan-type polysaccharides were recently isolated from its roots. Fructans are reserve carbohydrates that have important positive health effects and technological applications in the food industry. The objective of the present study was to isolate and characterize fructo-oligosaccharides (FOSs) from S. rebaudiana roots and in vitro adventitious root cultures and evaluate the potential prebiotic effect of these molecules. The in vitro adventitious root cultures were obtained using a roller bottle system. Chemical analyses (gas chromatography-mass spectrometry, (1)H nuclear magnetic resonance, and off-line electrospray ionization-mass spectrometry) revealed similar chemical properties of FOSs that were obtained from the different sources. The potential prebiotic effects of FOSs that were isolated from S. rebaudiana roots enhanced the growth of both bifidobacteria and lactobacilli, with strains specificity in their fermentation ability.
Selenized lactic acid bacteria (LAB) represent potentially safe and effective sources of selenium (Se), essential for human health, as lactic acid fermentation improves Se bioavailability and reduces its toxicity. LAB are generally recognized as safe (GRAS) and widely used in fermented dairy products. To facilitate selenized LAB implementation as a functional food, we developed and characterized new Se-enriched strains based on the food industry commercial strains Streptococcus thermophilus CCDM 144 and Enterococcus faecium CCDM 922A as representatives of two LAB genera. We evaluated Se bioaccumulation capacity, Se biotransformation and growth ability in the presence of different sodium selenite concentrations (0–50 mg/L), and antioxidant properties (2, 2-diphenyl-1-picrylhydrazyl (DPPH) method) and cell surface hydrophobicity between Se-enriched and parental strains in vitro. Sodium selenite addition did not negatively influence growth of either strain; thus, 50 mg/L was chosen as the optimal concentration based on strain accumulation capacity. Selenization improved the antioxidant properties of both strains and significantly increased their cell surface hydrophobicity (p < 0.05). To our knowledge, this represents the first report of Se-enriched strain hydrophobicity as well as the first on Se speciation in families Enterococcaceae and Streptococcaceae. Moreover, both tested strains demonstrated good potential for Se-enrichment, providing a foundation for further in vitro and in vivo studies to confirm the suitability of these Se-enriched strains for industrial applications.
The selenium (Se) enrichment of yeasts and lactic acid bacteria (LAB) has recently emerged as a novel concept; the individual health effects of these beneficial microorganisms are combined by supplying the essential micronutrient Se in a more bioavailable and less toxic form. This study investigated the bioavailability of Se in the strains Enterococcus faecium CCDM 922A (EF) and Streptococcus thermophilus CCDM 144 (ST) and their respective Se-enriched forms, SeEF and SeST, in a CD (SD-Sprague Dawley) IGS rat model. Se-enriched LAB administration resulted in higher Se concentrations in the liver and kidneys of rats, where selenocystine was the prevalent Se species. The administration of both Se-enriched strains improved the antioxidant status of the animals. The effect of the diet was more pronounced in the heart tissue, where a lower glutathione reductase content was observed, irrespective of the Se fortification in LAB. Interestingly, rats fed diets with EF and SeEF had higher glutathione reductase activity. Reduced concentrations of serum malondialdehyde were noted following Se supplementation. Diets containing Se-enriched strains showed no macroscopic effects on the liver, kidneys, heart, and brain and had no apparent influence on the basic parameters of the lipid metabolism. Both the strains tested herein showed potential for further applications as promising sources of organically bound Se and Se nanoparticles.
The ability of bacteria to adhere to the intestinal mucosa is a critical property necessary for the long-term colonization of the intestinal tract. This ability can be highly sensitive to the presence of prebiotics. However, limited data are available in this respect for beneficial bacteria such as probiotics or resident gut microbiota. We previously demonstrated that the presence of prebiotics may decrease adherence in several pre- and probiotic combinations. Thus, characterizing the interactions between numerous combinations involving different classes of pre- and probiotics can be crucial in identifying new synbiotics. Accordingly, here, we extend our prior analyses to evaluate the adhesion of five lactobacilli, six bifidobacteria, and one probiotic Escherichia coli strains, as commercial probiotics or promising probiotic candidates, together with the cariogenic Bifidobacterium dentium strain. As an in vitro intestinal mucosa model, Caco-2 and mucin-secreting HT29-MTX cells were co-cultured at 9:1 in the presence or absence of prebiotics. Commercial inulin-type fructooligosaccharide prebiotics Orafti® GR, Orafti® P95, and galactooligosaccharide-based prebiotic formula Vivinal®, including purified human milk oligosaccharides (HMOs) were added into the cultivation media as the sole sugar source (2.5% each). Adherence was tested using microtiter plates and was evaluated as the percentage of fluorescently labeled bacteria present in the wells after three washes. Consistent prebiotics-mediated enhanced adherence was observed only for the commercial probiotic strain E. coli O83. For the remaining strains, the presence of HMO or prebiotics Orafti® P95 or Orafti® GR decreased adherence, reaching statistical significance (p < 0.05) for three of out of eight (HMO) or five of out of 11 strains tested, respectively. Conversely, Vivinal® enhanced adhesion in six out of the 12 strains tested, and notably, it significantly attenuated the adherence of the cariogenic Bifidobacterium dentium Culture Collection of Dairy Microorganisms (CCDM) 318. To our knowledge, this represents the first report on the influence of commercial prebiotics and HMOs on the adhesion of the cariogenic Bifidobacterium sp. Vivinal® seems to be a promising prebiotic to be used in the formulation of synbiotics, supporting the adhesion of a wide range of probiotics, especially the strains B. bifidum BBV and BBM and the probiotic Escherichia coli O83.
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