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      Stimbiotic supplementation and xylose-rich carbohydrates modulate broiler’s capacity to ferment fibre

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          Abstract

          Stimbiotics are a new category of feed additives that can increase fibre fermentability by stimulating fibre-degrading microbiota in the gut. The aim of this study was to test, ex vivo, if the microbiota of broilers fed a stimbiotic are better able to ferment different xylose-rich substrates in an ileal and a caecal environment. The ileal and caecal contents from broiler chickens fed a stimbiotic or from a control group were used as an inoculum in the ex vivo fermentation experiment. Different xylose-rich substrates including monomeric xylose (XYL), XOS with DP 2 to 6 (XOS), short DP XOS of 2 to 3 (sDP-XOS), long DP XOS of 4 to 6 (lDP-XOS) and de-starched wheat bran (WB), were added to each ileal and caecal inoculum in fermentation vessels. Total gas, short-chain fatty acids (SCFA) production, bacterial quantification, and carbohydrate utilisation were monitored for 9 h post-inoculation. No significant interactions were observed in any of the parameters measured in either the ileal or caecal contents ( p > 0.05). Stimbiotic ileal inocula resulted in higher total gas ( p < 0.001) and volatile fatty acid (VFA) ( p < 0.001) production, increased numbers of Lactobacillus spp. ( p < 0.001), and decreased numbers of Enterococcus spp. ( p < 0.01) after 9 h regardless of the xylose-rich substrate added. Stimbiotic caecal inocula resulted in a higher ratio of VFA to branched-chain fatty acids (BCFAs) by up to +9% ( p < 0.05). Ileal microbiota were found to preferentially metabolise WB, while caecal microbiota favoured XOS substrates, particularly lDP-XOS. These results indicate that stimbiotics can promote the abundance of lactic acid bacteria involved in the establishment of fibre-degrading bacteria and VFA content in the gut, which could have beneficial effects on broiler performance. Further, ileal and caecal microbiota differ in their utilisation of different substrates which may impact the effectiveness of different stimbiotic products.

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          Determination of bacterial load by real-time PCR using a broad-range (universal) probe and primers set.

          The design and evaluation of a set of universal primers and probe for the amplification of 16S rDNA from the Domain Bacteria to estimate total bacterial load by real-time PCR is reported. Broad specificity of the universal detection system was confirmed by testing DNA isolated from 34 bacterial species encompassing most of the groups of bacteria outlined in Bergey's Manual of Determinative Bacteriology. However, the nature of the chromosomal DNA used as a standard was critical. A DNA standard representing those bacteria most likely to predominate in a given habitat was important for a more accurate determination of total bacterial load due to variations in 16S rDNA copy number and the effect of generation time of the bacteria on this number, since rapid growth could result in multiple replication forks and hence, in effect, more than one copy of portions of the chromosome. The validity of applying these caveats to estimating bacterial load was confirmed by enumerating the number of bacteria in an artificial sample mixed in vitro and in clinical carious dentine samples. Taking these parameters into account, the number of anaerobic bacteria estimated by the universal probe and primers set in carious dentine was 40-fold greater than the total bacterial load detected by culture methods, demonstrating the utility of real-time PCR in the analysis of this environment.
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            Lactate-utilizing bacteria, isolated from human feces, that produce butyrate as a major fermentation product.

            The microbial community of the human colon contains many bacteria that produce lactic acid, but lactate is normally detected only at low concentrations (<5 mM) in feces from healthy individuals. It is not clear, however, which bacteria are mainly responsible for lactate utilization in the human colon. Here, bacteria able to utilize lactate and produce butyrate were identified among isolates obtained from 10(-8) dilutions of fecal samples from five different subjects. Out of nine such strains identified, four were found to be related to Eubacterium hallii and two to Anaerostipes caccae, while the remaining three represent a new species within clostridial cluster XIVa based on their 16S rRNA sequences. Significant ability to utilize lactate was not detected in the butyrate-producing species Roseburia intestinalis, Eubacterium rectale, or Faecalibacterium prausnitzii. Whereas E. hallii and A. caccae strains used both D- and L-lactate, the remaining strains used only the d form. Addition of glucose to batch cultures prevented lactate utilization until the glucose became exhausted. However, when two E. hallii strains and one A. caccae strain were grown in separate cocultures with a starch-utilizing Bifidobacterium adolescentis isolate, with starch as the carbohydrate energy source, the L-lactate produced by B. adolescentis became undetectable and butyrate was formed. Such cross-feeding may help to explain the reported butyrogenic effect of certain dietary substrates, including resistant starch. The abundance of E. hallii in particular in the colonic ecosystem suggests that these bacteria play important roles in preventing lactate accumulation.
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              Microbiota, Gut Health and Chicken Productivity: What Is the Connection?

              Gut microbiota and its relationship to animal health and productivity in commercial broiler chickens has been difficult to establish due to high variability between flocks, which derives from plenty of environmental, nutritional, and host factors that influence the load of commensal and pathogenic microbes surrounding birds during their growth cycle in the farms. Chicken gut microbiota plays a key role in the maintenance of intestinal health through its ability to modulate host physiological functions required to maintain intestinal homeostasis, mainly through competitive exclusion of detrimental microorganisms and pathogens, preventing colonization and therefore decreasing the expense of energy that birds normally invest in keeping the immune system active against these pathogens. Therefore, a “healthy” intestinal microbiota implies energy saving for the host which translates into an improvement in productive performance of the birds. This review compiles information about the main factors that shape the process of gut microbiota acquisition and maturation, their interactions with chicken immune homeostasis, and the outcome of these interactions on intestinal health and productivity.
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                Author and article information

                Contributors
                URI : https://loop.frontiersin.org/people/2302583/overviewRole: Role: Role: Role: Role: Role: Role: Role: Role: Role: Role:
                Role: Role: Role: Role: Role: Role: Role: Role: Role: Role: Role: Role:
                Role: Role: Role: Role: Role: Role: Role: Role: Role: Role: Role: Role:
                Role: Role: Role: Role: Role: Role: Role: Role: Role: Role:
                URI : https://loop.frontiersin.org/people/2590017/overviewRole: Role: Role:
                Role: Role: Role: Role: Role: Role: Role: Role: Role: Role:
                Journal
                Front Microbiol
                Front Microbiol
                Front. Microbiol.
                Frontiers in Microbiology
                Frontiers Media S.A.
                1664-302X
                11 January 2024
                2023
                : 14
                : 1301727
                Affiliations
                [1] 1AB Vista , Wiltshire, United Kingdom
                [2] 2Alimetrics Research Ltd. , Espoo, Finland
                [3] 3School of Applied Sciences, University of Huddersfield , Huddersfield, United Kingdom
                Author notes

                Edited by: Karolina Skonieczna-Żydecka, Pomeranian Medical University, Poland

                Reviewed by: Lorenzo Nissen, University of Bologna, Italy; Paripok Phitsuwan, King Mongkut's University of Technology Thonburi, Thailand

                *Correspondence: Claire Davies, Claire.Davies@ 123456abvista.com
                Article
                10.3389/fmicb.2023.1301727
                10808361
                38274766
                4d82538c-c57d-4fcc-844a-b68846d1ce09
                Copyright © 2024 Davies, González-Ortiz, Rinttilä, Apajalahti, Alyassin and Bedford.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 25 September 2023
                : 20 December 2023
                Page count
                Figures: 2, Tables: 5, Equations: 0, References: 43, Pages: 11, Words: 8273
                Funding
                The author(s) declare that no financial support was received for the research, authorship, and/or publication of this article.
                Categories
                Microbiology
                Original Research
                Custom metadata
                Microorganisms in Vertebrate Digestive Systems

                Microbiology & Virology
                fibre fermentation,xylooligosaccharide,degree of polymerisation,ileal microbiota,caecal microbiota,scfa,ex vivo

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