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      Comprehensive Understanding of the Bacterial Populations and Metabolites Profile of Fermented Feed by 16S rRNA Gene Sequencing and Liquid Chromatography–Mass Spectrometry

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          Abstract

          The comprehensive bacterial populations and metabolites profile in fermented feed is unclear, which may have significant effects on the stability of fermented feed quality and animal gut health. In this study, 16S rRNA gene sequencing and liquid chromatography–mass spectrometry were used to explore the bacterial populations and metabolites profile in the fermented feed incubated with probiotics (MF) or without probiotics (SF). The probiotics were a combination of Lactobacillus salivarius, Bacillus subtilis, and Saccharomyces cerevisiae. The pH and lactic acid levels were higher in MF than in SF ( P < 0.05), while the total volatile fatty acid content was lower ( P < 0.05). Interestingly, after fermentation, the most abundant bacterial genus in MF was Enterococcus, rather than the added probiotics Lactobacillus or Bacillus. Weissella and a few potential pathogens ( Enterobacter, Escherichia-Shigella, and Pantoea) were dominant in SF ( P < 0.05). Metabolomics analysis identified 32 different metabolites in the two types of fermented feed. These metabolites enriched in MF, such as maleic acid, phenylacetic acid, ethyl linoleate, dihomo-gamma-linolenic acid, and L-theanine had potential antimicrobial activities. Conclusively, the addition of probiotics enriched a few potentially beneficial microbes and small molecular compounds with antimicrobial activities, and inhibited the potential pathogens in fermented feed.

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          Aspergillus oryzae GB-107 fermentation improves nutritional quality of food soybeans and feed soybean meals.

          This study evaluated the effect of fermentation on the nutritional quality of food-grade soybeans and feed-grade soybean meals. Soybeans and soybean meals were fermented by Aspergillus oryzae GB-107 in a bed-packed solid fermentor for 48 hours. After fermentation, their nutrient contents as well as trypsin inhibitor were measured and compared with those of raw soybeans and soybean meals. Proteins were extracted from fermented and non-fermented soybeans and soybean meals, and the peptide characteristics were evaluated after electrophoresis. Fermented soybeans and fermented soybean meals contained 10% more (P 60 kDa) (P 60 kDa), whereas 22.1% of peptides in soybean meal were large-size (>60 kDa). Collectively, fermentation increased protein content, eliminated trypsin inhibitors, and reduced peptide size in soybeans and soybean meals. These effects of fermentation might make soy foods more useful in human diets as a functional food and benefit livestock as a novel feed ingredient.
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            Antimicrobial and Probiotic Properties of Yeasts: From Fundamental to Novel Applications

            The yeasts constitute a large and heterogeneous group of microorganisms that are currently attracting increased attention from scientists and industry. Numerous and diverse biological activities make them promising candidates for a wide range of applications not limited to the food sector. In addition to their major contribution to flavor development in fermented foods, their antagonistic activities toward undesirable bacteria, and fungi are now widely known. These activities are associated with their competitiveness for nutrients, acidification of their growth medium, their tolerance of high concentrations of ethanol, and release of antimicrobial compounds such as antifungal killer toxins or “mycocins” and antibacterial compounds. While the design of foods containing probiotics (microorganisms that confer health benefits) has focused primarily on Lactobacillus and Bifidobacterium, the yeast Saccharomyces cerevisiae var. boulardii has long been known effective for treating gastroenteritis. In this review, the antimicrobial activities of yeasts are examined. Mechanisms underlying this antagonistic activity as well as recent applications of these biologically active yeasts in both the medical and veterinary sectors are described.
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              Health-beneficial effects of probiotics: Its mode of action.

              It is now widely recognized that probiotics have health-beneficial effects on humans and animals. Probiotics should survive in the intestinal tract to exert beneficial effects on the host's health. To keep a sufficient level of probiotic bacteria in the gastrointestinal tract, a shorter interval between doses may be required. Although adherence to the intestinal epithelial cell and mucus is not a universal property of probiotics, high ability to adhere to the intestinal surface might strongly interfere with infection of pathogenic bacteria and regulate the immune system. The administration of probiotic Lactobacillus stimulated indigenous Lactobacilli and the production of short-chain fatty acids. This alteration of the intestinal environment should contribute to maintain the host's health. The immunomodulatory effects of probiotics are related to important parts of their beneficial effects. Probiotics may modulate the intestinal immune response through the stimulation of certain cytokine and IgA secretion in intestinal mucosa. The health-beneficial effects, in particular the immunomodulation effect, of probiotics depend on the strain used. Differences in indigenous intestinal microflora significantly alter the magnitude of the effects of a probiotic. Specific probiotic strains suitable for each animal species and their life stage as well as each individual should be found.
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                Author and article information

                Journal
                Metabolites
                Metabolites
                metabolites
                Metabolites
                MDPI
                2218-1989
                21 October 2019
                October 2019
                : 9
                : 10
                : 239
                Affiliations
                [1 ]Laboratory of Gastrointestinal Microbiology, Jiangsu Key Laboratory of Gastrointestinal Nutrition and Animal Health, College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China; jinwei@ 123456njau.edu.cn (W.J.); 2016105045@ 123456njau.edu.cn (Z.Z.); 2016805092@ 123456njau.edu.cn (K.Z.); xueyanfeng1990@ 123456163.com (Y.X.); 2017105050@ 123456njau.edu.cn (F.X.)
                [2 ]National Center for International Research on Animal Gut Nutrition, Nanjing Agricultural University, Nanjing 210095, China
                [3 ]National Experimental Teaching Demonstration Center of Animal Science, Nanjing Agricultural University, Nanjing 210095, China
                Author notes
                Author information
                https://orcid.org/0000-0002-0089-9314
                Article
                metabolites-09-00239
                10.3390/metabo9100239
                6835224
                31640120
                bbdf23a0-c2e0-471c-9403-7323194f57ee
                © 2019 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 17 September 2019
                : 15 October 2019
                Categories
                Article

                fermented feed,bacterial population,small molecular metabolites,lactobacillus salivarius,bacillus subtilis,saccharomyces cerevisiae

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