Effect of in vitro cultivation on human gut microbiota composition using 16S rDNA amplicon sequencing and metabolomics approach.

Gut microbiota (GM) plays many key functions and helps maintain the host's health. Consequently, the development of GM cultivation under in vitro stimulating physiological conditions has gained extreme interest in different fields. In this study, we evaluated the impact of four culture media: Gut Microbiota Medium (GMM), Schaedler Broth (SM), Fermentation Medium (FM), and Carbohydrate Free Basal Medium (CFBM) on preserving the biodiversity and metabolic activity of human GM in batch in vitro cultures using PMA treatment coupled with 16S rDNA sequencing (PMA-seq) and LC-HR-MS/MS untargeted metabolomics supplemented with GC-MS SCFA profiling. Before the experiments, we determined the possibility of using the pooled faecal samples (MIX) from healthy donors (n = 15) as inoculum to reduce the number of variables and ensure the reproducibility of in vitro cultivation tests. Results showed the suitability of pooling faecal samples for in vitro cultivation study. Non-cultured MIX inoculum was characterized by higher α-diversity (Shannon effective count, and Effective microbial richness) compared to inocula from individual donors. After 24 h of cultivation, a significant effect of culture media composition on GM taxonomic and metabolomic profiles was observed. The SM and GMM had the highest α-diversity (Shannon effective count). The highest number of core ASVs (125) shared with non-cultured MIX inoculum and total SCFAs production was observed in the SM. These results might contribute to the development of standardized protocols for human GM in vitro cultivation by preventing methodological bias in the data.

Efficacy and Safety Assessment of Microbiological Feed Additive for Chicken Broilers in Tolerance Studies.

INTRODUCTION: One aim of the study was to evaluate the impact when added to feed of the two potentially probiotic strains of lactic acid bacteria (LAB) Lactobacillus plantarum K KKP 593/p and Lactobacillus rhamnosus KKP 825 on production performance, health, and the composition of gut microbiota. The complementary aim was to assess the safety of these strains in broiler rearing. MATERIAL AND METHODS: A total of 500 one-day-old Ross 308 chicks were divided into four groups. The experimental factor was the admixture of bacterial preparation to the feed at different doses: the recommended maximum dose, a dose ten times higher, the recommended minimum dose, and a zero dose for the control group not receiving bacteria. RESULTS: Addition of bacteria to the diets did not have a significant effect on the final body weight, final body weight gain, nor total feed intake or feed conversion. However, lactic acid bacteria had a positive effect on chicken health. Mortality among chickens fed with LAB was reduced. Moreover, LAB feeding inhibited the growth of Salmonella spp. and Clostridium perfringens in the intestines. There were no significant differences in chicken performance by dose of bacteria in the feed. The group dosed with LAB ten times higher than the recommended maximum did not demonstrate changes in biochemical or haematological parameters of blood compared to the remaining groups. CONCLUSION: Feeding chicken broilers with two potentially probiotic LAB strains is safe and impacts animal health positively.

Selection of lactic acid bacteria strains for the hydrolysis of allergenic proteins of wheat flour.

BACKGROUND: Wheat flour is one of the most common causative agents of food allergy. The study presents the selection and characterization of lactic acid bacteria (LAB) strains capable of hydrolyzing/modifying allergenic proteins of wheat flour. Hydrolysis of wheat proteins was determined with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting with sera from patients with food allergy to gluten. RESULTS: The analysis of electrophoretic profiles of protein extracted from sourdough shows the capability of selected LAB strains for proteolytic degradation of wheat proteins that belong to two factions: albumin/globulin (hydrolysis of 13 polypeptides with a molecular weight between 103 and 22 kDa); and gliadin (seven polypeptides with a molecular weight between 39 and 24 kDa). All analyzed strains were capable of hydrolyzing some IgE-binding epitopes of wheat allergens. The lack of such changes in control samples indicates that they were induced rather by the proteolytic activity of bacterial strains than endogenous enzymes of wheat flour. The gluten proteins were susceptible to hydrolysis by sequential digestion with pepsin and trypsin. CONCLUSION: The selected strains exhibit proteolytic activity, which leads to a reduction in allergenicity of wheat sourdoughs. These strains may be applied as specific starter cultures to prepare bakery products of special nutritional use. © 2015 Society of Chemical Industry.

Assortment of carbon sources in medium for Yarrowia lipolytica lipase production: A statistical approach.

Glycerol is considered an important renewable feedstock as well as an undesirable side-product of biodiesel production. The aim of this study was to determine whether supplementing a culture medium with a combination of three different carbon sources (olive oil, glucose and glycerol) would optimize lipase production by the yeast Yarrowia lipolytica. The optimization experiments were conducted with a statistical approach using the mixture design. Analysis of the response surface revealed that it would be possible to compose a medium in which both an an extracellular lipase activity of 0.1 U/mL and up to 37.5 g/L of pure glycerol could be obtained. An YPO-Gl30 medium consisting of 30 g/L glycerol and 19.2 mL/L olive oil was selected for further investigation. Although a high biomass yield was found in all cultures, the glycerol content of the YPO-Gl30 medium slightly influenced yeast growth, but it did not prolong the duration of the lag phase. The hydrolytic activity of the extracellular lipases produced in YPO-Gl30 medium was satisfactory.