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      Composition and Diversity of the Fecal Microbiome and Inferred Fecal Metagenome Does Not Predict Subsequent Pneumonia Caused by Rhodococcus equi in Foals

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          Abstract

          In equids, susceptibility to disease caused by Rhodococcus equi occurs almost exclusively in foals. This distribution might be attributable to the age-dependent maturation of immunity following birth undergone by mammalian neonates that renders them especially susceptible to infectious diseases. Expansion and diversification of the neonatal microbiome contribute to development of immunity in the gut. Moreover, diminished diversity of the gastrointestinal microbiome has been associated with risk of infections and immune dysregulation. We thus hypothesized that varying composition or reduced diversity of the intestinal microbiome of neonatal foals would contribute to increased susceptibility of their developing R. equi pneumonia. The composition and diversity indices of the fecal microbiota at 3 and 5 weeks of age were compared among 3 groups of foals: 1) foals that subsequently developed R. equi pneumonia after sampling; 2) foals that subsequently developed ultrasonographic evidence of pulmonary abscess formation or consolidation but not clinical signs (subclinical group); and, 3) foals that developed neither clinical signs nor ultrasonographic evidence of pulmonary abscess formation or consolidation. No significant differences were found among groups at either sampling time, indicating absence of evidence of an influence of composition or diversity of the fecal microbiome, or predicted fecal metagenome, on susceptibility to subsequent R. equi pneumonia. A marked and significant difference identified between a relatively short interval of time appeared to reflect ongoing adaptation to transition from a milk diet to a diet including available forage (including hay) and access to concentrate fed to the mare.

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          Most cited references37

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          Innate immunity of the newborn: basic mechanisms and clinical correlates.

          Ofer Levy (2007)
          The fetus and newborn face a complex set of immunological demands, including protection against infection, avoidance of harmful inflammatory immune responses that can lead to pre-term delivery, and balancing the transition from a sterile intra-uterine environment to a world that is rich in foreign antigens. These demands shape a distinct neonatal innate immune system that is biased against the production of pro-inflammatory cytokines. This bias renders newborns at risk of infection and impairs responses to many vaccines. This Review describes innate immunity in newborns and discusses how this knowledge might be used to prevent and treat infection in this vulnerable population.
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            Rapid fucosylation of intestinal epithelium sustains host-commensal symbiosis in sickness

            Systemic infection induces conserved physiological responses that include both resistance and ‘tolerance of infection’ mechanisms 1 . Temporary anorexia associated with an infection is often beneficial 2,3 reallocating energy from food foraging towards resistance to infection 4 or depriving pathogens of nutrients 5 . It imposes, however, a stress on intestinal commensals, as they also experience reduced substrate availability and impacting host fitness due to the loss of caloric intake and colonization resistance (protection from additional infections) 6 . We hypothesized that the host might utilize internal resources to support the gut microbiota during the acute phase of the disease. Here we show that systemic exposure to Toll-like receptor (TLR) ligands causes rapid α1,2-fucosylation of the small intestine epithelial cells (IEC), which requires sensing of TLR agonists and production of IL-23 by dendritic cells, activation of innate lymphoid cells and expression of α1,2-Fucosyltransferase-2 (Fut2) by IL-22-stimulated IECs. Fucosylated proteins are shed into the lumen and fucose is liberated and metabolized by the gut microbiota, as shown by reporter bacteria and community-wide analysis of microbial gene expression. Fucose affects the expression of microbial metabolic pathways and reduces the expression of bacterial virulence genes. It also improves host tolerance of the mild pathogen Citrobacter rodentium. Thus, rapid IEC fucosylation appears to be a protective mechanism that utilizes the host's resources to maintain host-microbial interactions during pathogen-induced stress.
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              Taxonomic Classification of Bacterial 16S rRNA Genes Using Short Sequencing Reads: Evaluation of Effective Study Designs

              Massively parallel high throughput sequencing technologies allow us to interrogate the microbial composition of biological samples at unprecedented resolution. The typical approach is to perform high-throughout sequencing of 16S rRNA genes, which are then taxonomically classified based on similarity to known sequences in existing databases. Current technologies cause a predicament though, because although they enable deep coverage of samples, they are limited in the length of sequence they can produce. As a result, high-throughout studies of microbial communities often do not sequence the entire 16S rRNA gene. The challenge is to obtain reliable representation of bacterial communities through taxonomic classification of short 16S rRNA gene sequences. In this study we explored properties of different study designs and developed specific recommendations for effective use of short-read sequencing technologies for the purpose of interrogating bacterial communities, with a focus on classification using naïve Bayesian classifiers. To assess precision and coverage of each design, we used a collection of ∼8,500 manually curated 16S rRNA gene sequences from cultured bacteria and a set of over one million bacterial 16S rRNA gene sequences retrieved from environmental samples, respectively. We also tested different configurations of taxonomic classification approaches using short read sequencing data, and provide recommendations for optimal choice of the relevant parameters. We conclude that with a judicious selection of the sequenced region and the corresponding choice of a suitable training set for taxonomic classification, it is possible to explore bacterial communities at great depth using current technologies, with only a minimal loss of taxonomic resolution.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                25 August 2015
                2015
                : 10
                : 8
                : e0136586
                Affiliations
                [1 ]Department of Large Animal Clinical Sciences, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, Texas, United States of America
                [2 ]Department of Small Animal Clinical Sciences, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, Texas, United States of America
                [3 ]Molecular Research DNA Laboratory (MR DNA), Shallowater, Texas, United States of America
                [4 ]6666 Ranch, Guthrie, Texas, United States of America
                The University of Melbourne, AUSTRALIA
                Author notes

                Competing Interests: 1) This study was supported by a grant from Boehringer Ingelheim; 2) Scot Dowd is an employee of Molecular Research DNA Laboratory and receives a salary from Molecular Research DNA Laboratory, Shallowater, TX; and, 3) Glenn P. Blodgett is employed by 6666 Ranch. There are no patents, products in development, or marketed products to declare. This does not alter the authors' adherence to all the PLoS ONE policies on sharing data and materials.

                Conceived and designed the experiments: CW-C NDC MKC CA JS. Performed the experiments: CW-C SED. Analyzed the data: CW-C NDC JS SED CMM. Contributed reagents/materials/analysis tools: JS GPB. Wrote the paper: CW-C NDC JS MKC CA SED GPB.

                Article
                PONE-D-15-14430
                10.1371/journal.pone.0136586
                4549325
                26305682
                b5a3babb-aca2-4226-a73b-ff8e8fde1ca9
                Copyright @ 2015

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited

                History
                : 7 April 2015
                : 5 August 2015
                Page count
                Figures: 8, Tables: 1, Pages: 19
                Funding
                This work was supported by a grant from Boehringer Ingelheim (Grant number 01072014) http://www.boehringeringelheim.com/. Additional funding was provided by the Link Equine Research Endowment. Fecal sample collection was made possible by a grant from the American Quarter Horse Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Scot E. Dowd is employed by Molecular Research DNA Laboratory (MR DNA). Molecular Research DNA Laboratory (MR DNA) provided support in the form of salary for author SED, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific Cover Letter 2 role of this author is articulated in the ‘author contributions’ section. Glenn P. Blodgett is employed by 6666 Ranch. 6666 Ranch provided support in the form of salary for author GPB, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific role of this author is articulated in the ‘author contributions’ section.
                Categories
                Research Article
                Custom metadata
                Data are available from the NCBI GenBank database submission number SRP060764.

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