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The clinical course of the 2019 coronavirus disease (COVID-19) is variable and to a substantial degree still unpredictable, especially in persons who have neither been vaccinated nor recovered from previous infection. We hypothesized that disease progression and inflammatory responses were associated with alterations in the microbiome and metabolome. To test this, we integrated metagenome, metabolome, cytokine, and transcriptome profiles of longitudinally collected samples from hospitalized COVID-19 patients at the beginning of the pandemic (before vaccines or variants of concern) and non-infected controls, and leveraged detailed clinical information and post-hoc confounder analysis to identify robust within- and cross-omics associations. Severe COVID-19 was directly associated with a depletion of potentially beneficial intestinal microbes mainly belonging to Clostridiales, whereas oropharyngeal microbiota disturbance appeared to be mainly driven by antibiotic use. COVID-19 severity was also associated with enhanced plasma concentrations of kynurenine, and reduced levels of various other tryptophan metabolites, lysophosphatidylcholines, and secondary bile acids. Decreased abundance of Clostridiales potentially mediated the observed reduction in 5-hydroxytryptophan levels. Moreover, altered plasma levels of various tryptophan metabolites and lower abundances of Clostridiales explained significant increases in the production of IL-6, IFNγ and/or TNFα. Collectively, our study identifies correlated microbiome and metabolome alterations as a potential contributor to inflammatory dysregulation in severe COVID-19.Graphical Abstract

Evaluating metagenomic software is key for optimizing metagenome interpretation and focus of the Initiative for the Critical Assessment of Metagenome Interpretation (CAMI). The CAMI II challenge engaged the community to assess methods on realistic and complex datasets with long- and short-read sequences, created computationally from around 1,700 new and known genomes, as well as 600 new plasmids and viruses. Here we analyze 5,002 results by 76 program versions. Substantial improvements were seen in assembly, some due to long-read data. Related strains still were challenging for assembly and genome recovery through binning, as was assembly quality for the latter. Profilers markedly matured, with taxon profilers and binners excelling at higher bacterial ranks, but underperforming for viruses and Archaea. Clinical pathogen detection results revealed a need to improve reproducibility. Runtime and memory usage analyses identified efficient programs, including top performers with other metrics. The results identify challenges and guide researchers in selecting methods for analyses., Nature Methods, 19 (8), ISSN:1548-7105, ISSN:1548-7091

Summary Background Rheumatoid arthritis is a chronic inflammatory autoimmune disease that is associated with reduced life expectancy. The disease is heritable and an extensive repertoire of genetic variants have been identified. The gut microbiota might represent an environmental risk factor for rheumatoid arthritis. We aimed to assess whether known rheumatoid arthritis risk alleles were associated with the gut microbiota in a large population who do not have rheumatoid arthritis. Methods In this cross-sectional study done in the UK and Switzerland, we used genotyping and microbiota data from previous studies of the TwinsUK cohort, excluding participants who had ever had a diagnosis of rheumatoid arthritis, as well as their unaffected co-twins. We used blood samples for genotyping and stool samples for the assessment of the gut microbiota. We generated a polygenic risk score (PRS) for rheumatoid arthritis in 1650 TwinsUK participants without the disease, based on 233 GWAS-identified single nucleotide polymorphisms associated with rheumatoid arthritis. We validated the PRS using logistic regression against rheumatoid arthritis diagnosis in 2686 UK Biobank individuals with a confirmed diagnosis of rheumatoid arthritis. Amplicon sequence variants (ASVs) were generated from 16S rRNA gene sequencing of stool samples and assessed for association with the PRS for rheumatoid arthritis. We validated the findings in an independent sample comprised of first-degree relatives of patients with rheumatoid arthritis from the SCREEN-RA cohort. Differential abundance of ASVs present in more than 5% of samples, grouped by ASV taxon annotation, against the rheumatoid arthritis PRS as a continuous variable was assessed using fixed-effects covariates. To account for multiple testing, the false discovery rate calculation was applied to all p values to generate q values, with a significance threshold of 0·05 determined a priori. Findings We found that presence of Prevotella spp were positively associated with the rheumatoid arthritis PRS in TwinsUK participants (q

Prevotella copri is a prevalent inhabitant of the human gut and has been associated with plant‐rich diet consumption and diverse health states. The underlying genetic basis of these associations remains enigmatic due to the lack of genetic tools. Here, we developed a novel versatile genetic toolbox for rapid and efficient genetic insertion and allelic exchange applicable to P. copri strains from multiple clades. Enabled by the genetic platform, we systematically investigated the specificity of polysaccharide utilization loci (PULs) and identified four highly conserved PULs for utilizing arabinan, pectic galactan, arabinoxylan, and inulin, respectively. Further genetic and functional analysis of arabinan utilization systems illustrate that P. copri has evolved two distinct types of arabinan‐processing PULs (PULAra) and that the type‐II PULAra is significantly enriched in individuals consuming a vegan diet compared to other diets. In summary, this genetic toolbox will enable functional genetic studies for P. copri in future., A novel conjugation‐based gene insertion/excision system allowing genetic manipulation of the previously intractable bacterium Prevotella copri provides insight into human diet effects on commensal bacterial metabolism.

Prevotella copriis a prevalent inhabitant of the human gut and has been associated with plant-rich diet consumption and diverse health states. The underlying genetic basis of these associations remains enigmatic due to the lack of genetic tools. Here, we developed a novel versatile genetic toolbox for rapid and efficient genetic insertion and allelic exchange applicable toP. copristrains from multiple clades. Enabled by the genetic platform, we systematically investigated the specificity of polysaccharide utilization loci (PULs), and identified four highly conserved PULs for utilizing arabinan, pectic galactan, arabinoxylan and inulin, respectively. Further genetic and functional analysis of arabinan utilization systems illustrate thatP. coprihas evolved two distinct types of arabinan-processing PULs (PULAra) and that the type-II PULArais significantly enriched in individuals consuming a vegan diet compared to other diets. In summary, this genetic toolbox will enable functional genetic studies forP. copriin the future.

Computational methods are key in microbiome research, and obtaining a quantitative and unbiased performance estimate is important for method developers and applied researchers. For meaningful comparisons between methods, to identify best practices and common use cases, and to reduce overhead in benchmarking, it is necessary to have standardized datasets, procedures and metrics for evaluation. In this tutorial, we describe emerging standards in computational meta-omics benchmarking derived and agreed upon by a larger community of researchers. Specifically, we outline recent efforts by the Critical Assessment of Metagenome Interpretation (CAMI) initiative, which supplies method developers and applied researchers with exhaustive quantitative data about software performance in realistic scenarios and organizes community-driven benchmarking challenges. We explain the most relevant evaluation metrics for assessing metagenome assembly, binning and profiling results, and provide step-by-step instructions on how to generate them. The instructions use simulated mouse gut metagenome data released in preparation for the second round of CAMI challenges and showcase the use of a repository of tool results for CAMI datasets. This tutorial will serve as a reference for the community and facilitate informative and reproducible benchmarking in microbiome research. This tutorial explains how to evaluate and benchmark metagenome assembly, binning and profiling methods using standards and software provided by the CAMI initiative.

In viral infections often multiple related viral strains are present, due to coinfection or within-host evolution. We describe Haploflow, a de Bruijn graph-based assembler for de novo genome assembly of viral strains from mixed sequence samples using a novel flow algorithm. We assessed Haploflow across multiple benchmark data sets of increasing complexity, showing that Haploflow is faster and more accurate than viral haplotype assemblers and generic metagenome assemblers not aiming to reconstruct strains. Haplotype reconstructed high-quality strain-resolved assemblies from clinical HCMV samples and SARS-CoV-2 genomes from wastewater metagenomes identical to genomes from clinical isolates.

CD81 plays a role in a variety of physiological and pathological processes. Recent structural analysis of CD81 indicates that it contains an intramembrane cholesterol-binding pocket and that interaction with cholesterol may regulate a conformational switch in the extracellular domain of CD81. Therefore, CD81 possesses a potential cholesterol sensing mechanism; however, its relevance for protein function is thus far unknown. In this study we investigate CD81 cholesterol sensing in the context of its activity as a receptor for hepatitis C virus. Structure-led mutagenesis of the cholesterol-binding pocket reduced CD81-cholesterol association, but had disparate effects on HCV, both reducing and enhancing CD81 receptor activity. We reasoned that this could be explained by alterations in the consequences of cholesterol binding. To investigate this further we performed molecular dynamic simulations of CD81 with and without cholesterol; this identified an allosteric mechanism by which cholesterol binding regulates the conformation of CD81. To test this, we designed further mutations to force CD81 into either the open (cholesterol unbound) or closed (cholesterol bound) conformation. The open mutant of CD81 exhibited reduced receptor activity whereas the closed mutant was enhanced. These data are consistent with cholesterol switching CD81 between a receptor active and inactive state. CD81 interactome analysis also suggests that conformational switching may modulate the assembly of CD81-partner networks. This work furthers our understanding of the molecular mechanism of CD81 cholesterol sensing, how this relates to HCV entry and CD81's function as a molecular scaffold; these insights are relevant to CD81's varied roles in health and disease.

In their outstanding study, Wei et al 1 compared the intestinal microbiota (IM) of well-defined autoimmune hepatitis (AIH) patients to healthy controls. They reported significantly reduced IM alpha diversity and disparate IM compositional heterogeneity (beta diversity) with enrichment of Streptococcus , Veillonella , Klebsiella and Lactobacillus in AIH. The excellent achievement of this study is to have characterised the IM of untreated patients with AIH. However, differentiation between IM changes caused by AIH in particular and by chronic liver disease (CLD) in general was not possible because this study lacked a control group with CLD other than AIH. In a pilot study, we analysed the IM of patients with CLD of mixed aetiology in comparison to healthy controls to clarify alterations across different stages of CLD (table 1 and online supplementary data). In contrast to the study by Wei et al , patients with recent intake of antibiotics were excluded. Additionally, only patients on proton pump inhibitors (PPIs), which are frequently taken by patients with CLD, were included to avoid a bias due to mixed PPI use, because PPI can alter the IM, in particular, the abundance of Veillonellaceae and Streptococcaceae.2 Based on investigations of normalised relative reads (n-RR) per IM taxon as well as IM alpha-diversity and beta-diversity analyses, we detected Veillonella …

Diverse microbial signatures within the intestinal microbiota have been associated with intestinal and systemic inflammatory diseases, but whether these candidate microbes actively modulate host phenotypes or passively expand within the altered microbial ecosystem is frequently not known. Here we demonstrate that colonization of mice with a member of the genus Prevotella, which has been previously associated to colitis in mice, exacerbates intestinal inflammation. Our analysis revealed that Prevotella intestinalis alters composition and function of the ecosystem resulting in a reduction of short-chain fatty acids, specifically acetate, and consequently a decrease in intestinal IL-18 levels during steady state. Supplementation of IL-18 to Prevotella-colonized mice was sufficient to reduce intestinal inflammation. Hence, we conclude that intestinal Prevotella colonization results in metabolic changes in the microbiota, which reduce IL-18 production and consequently exacerbate intestinal inflammation, and potential systemic autoimmunity.

Summary The complexity of host-associated microbial ecosystems requires host-specific reference catalogs to survey the functions and diversity of these communities. We generate a comprehensive resource, the integrated mouse gut metagenome catalog (iMGMC), comprising 4.6 million unique genes and 660 metagenome-assembled genomes (MAGs), many (485 MAGs, 73%) of which are linked to reconstructed full-length 16S rRNA gene sequences. iMGMC enables unprecedented coverage and taxonomic resolution of the mouse gut microbiota; i.e., more than 92% of MAGs lack species-level representatives in public repositories (, Graphical Abstract, Highlights • Large-scale metagenomic assembly uncovers hundreds of mouse microbiome species • Most microbes found in the mouse gut are unique to the ecosystem • Integration of gene catalog and microbial genomes creates a comprehensive resource • Linking of 16S rRNA genes with metagenome-assembled genomes allows new applications, Gene catalogs and genome references facilitate taxonomic and functional annotation of sequencing data. Through the combination of data from laboratory and wild mice, Lesker et al. create a comprehensive resource, the integrated mouse gut metagenome catalog, to characterize the microbial ecosystem in the murine gut.

Periods of fasting and refeeding may reduce cardiometabolic risk elevated by Western diet. We show that in hypertensive metabolic syndrome (MetS) patients (n=35), a 5-day fast followed by a modified DASH diet (Dietary Approach to Stop Hypertension) reduced systolic blood pressure (SBP), antihypertensive medication need, and body-mass index (BMI) at three months post intervention compared to a modified DASH diet alone (n=36). Fasting altered the gut microbiome, impacting bacterial taxa and gene modules associated with short-chain fatty acid production. Cross-system analyses revealed a positive correlation of circulating mucosa-associated invariant T (MAIT) cells, non-classical monocytes and CD4+ effector T cells with SBP. Furthermore, regulatory T cells (Tregs) positively correlated with BMI and weight. Machine learning could predict sustained SBP-responsiveness within the fasting group from baseline immunome data, identifying CD8+ effector T cells, Th17 cells and Tregs as important contributors to the model. The high-resolution multi-omics data highlights fasting as a promising non-pharmacological intervention in MetS.

Our knowledge about the gut microbiota of pigs is still scarce, despite the importance of these animals for biomedical research and agriculture. Here, we present a collection of cultured bacteria from the pig gut, including 110 species across 40 families and nine phyla. We provide taxonomic descriptions for 22 novel species and 16 genera. Meta-analysis of 16S rRNA amplicon sequence data and metagenome-assembled genomes reveal prevalent and pig-specific species within Lactobacillus, Streptococcus, Clostridium, Desulfovibrio, Enterococcus, Fusobacterium, and several new genera described in this study. Potentially interesting functions discovered in these organisms include a fucosyltransferase encoded in the genome of the novel species Clostridium porci, and prevalent gene clusters for biosynthesis of sactipeptide-like peptides. Many strains deconjugate primary bile acids in in vitro assays, and a Clostridium scindens strain produces secondary bile acids via dehydroxylation. In addition, cells of the novel species Bullifex porci are coccoidal or spherical under the culture conditions tested, in contrast with the usual helical shape of other members of the family Spirochaetaceae. The strain collection, called ‘Pig intestinal bacterial collection’ (PiBAC), is publicly available at www.dsmz.de/pibac and opens new avenues for functional studies of the pig gut microbiota., The authors present a public collection of 117 bacterial isolates from the pig gut, including the description of 38 novel taxa. Interesting functions discovered in these organisms include a new fucosyltransferease and sactipeptide-like molecules encoded by biosynthetic gene clusters.

Prevotella spp. form one of the most dominant bacterial genera in the human gut. Multiple Prevotella spp. can co-exist in some but not all individuals consuming plant-based diets. To functionally investigate the relationship between Prevotella competition and diet, we isolated several novel Prevotella spp. from the mouse gut, performed genome and metatranscriptome analysis as well as a series of competition experiments under different dietary conditions. These experiments revealed that diverse dominant Prevotella species compete for similar metabolic niches in vivo, which is linked to the upregulation of specific Polysaccharide Utilization Loci (PULs). Complex plant-derived polysaccharides are required, and particularly arabinoxylan is sufficient, for the expansion of Prevotella spp. The most dominant Prevotella species encodes a specific tandem-repeat trsusC/D PUL, which is conserved in highly abundant Prevotella copri strains further expanded in humans consuming a vegan diet. Our data suggest efficient (arabino)xylan-utilization as a factor contributing to Prevotella dominance.

BackgroundRheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease associated with reduced life expectancy. It is heritable and an extensive repertoire of genetic variants have been identified. The gut microbiota may represent an environmental risk factor for RA. Indeed, Prevotella copri is a candidate keystone species, but whether it lies on the causal pathway for disease or is simply a bystander reflecting host-genetic predisposition to RA, remains to be determined. The study of disease-microbiota associations may be confounded by the presence of the disease of interest or by its treatment. To circumvent this issue, we assessed whether known RA risk alleles were associated with the gut microbiota, in a large who do not have RA.MethodsBlood and stool acquired from volunteers from TwinsUK were used for genotyping and assessment of the gut microbiota, respectively. A weighted polygenic risk score (PRS) for RA was calculated in 1650 unaffected twins from the TwinsUK cohort, based on 233 published GWAS-identified published RA associated single nucleotide polymorphisms (SNPs). Amplicon sequence variants (ASVs) were generated from 16S rRNA sequencing of stool samples and assessed for association with RA PRS. Confirmation of findings was performed using an independent sample comprised of first-degree relatives of RA patients from the SCREEN-RA cohort (n=133).FindingsWe found that Prevotella spp was positively associated with RA PRS in TwinsUK participants (p.adjPrevotella spp. with pre-clinical RA phases was also demonstrated (p.adj=0.021).InterpretationPrevotella in the gut microbiota is associated with RA genotype in the absence of RA, as well as in subjects at high risk of developing RA. This work suggests that host genotype is associated with microbiota profile prior to disease onset.Research in contextEvidence before this study?Prevotella copri is increased within the gut microbiota in rheumatoid arthritis (RA) patients, predominantly those with early disease, before treatment is initiated. Prevotellaceae (family) has been demonstrated to be increased in pre-clinical RA cases compared to controls. Prevotella copri is posited to be an inflammatory driver, contributing to RA pathology by promoting a pro-inflammatory cytokine milieu.What does this study add?This study is the first to demonstrate that, in a large human cohort, carrying genetic risk factors for RA is associated with a higher abundance of Prevotella spp. in the absence of RA.How might this impact on clinical practice or future developmentsThis work suggests that any potential causal role of Prevotella spp. occurs very early in disease development. Strategies to intervene should target early or asymptomatic at-risk groups

We are grateful to Dr Inamo1 for his interest in our article. We agree and acknowledged in our manuscript that our study design did not allow us to assess a causal association. Our study described an increased relative abundance in Prevotella spp. in individuals in ‘preclinical rheumatoid arthritis (RA) stages’ using participants enrolled in a first-degree relatives of patients with RA (FDR-RA) cohort.2 The microbiota of individuals in preclinical RA stages were significantly altered compared with FDR-RA controls. In particular, bacteria of the Prevotellaceae family and associated taxa were enriched among individuals in preclinical stages of RA. Dr …

The composition of the intestinal microbiota influences the outcome of enteric infections in human and mice. However, the role of specific members and their metabolites contributing to disease severity is largely unknown. Using isogenic mouse lines harboring distinct microbiota communities, we observed highly variable disease kinetics of enteric Citrobacter rodentium colonization after infection. Transfer of communities from susceptible and resistant mice into germ-free mice verified that the varying susceptibilities are determined by microbiota composition. The strongest differences in colonization were observed in the cecum and could be maintained in vitro by coculturing cecal bacteria with C. rodentium. Cohousing of animals as well as the transfer of cultivable bacteria from resistant to susceptible mice led to variable outcomes in the recipient mice. Microbiome analysis revealed that a higher abundance of butyrate-producing bacteria was associated with the resistant phenotype. Quantification of short-chain fatty acid (SCFA) levels before and after infection revealed increased concentrations of acetate, butyrate and propionate in mice with delayed colonization. Addition of physiological concentrations of butyrate, but not of acetate and/or propionate strongly impaired growth of C. rodentium in vitro. In vivo supplementation of susceptible, antibiotic-treated and germ-free mice with butyrate led to the same level of protection, notably only when cecal butyrate concentration reached a concentration higher than 50 nmol/mg indicating a critical threshold for protection. In the recent years, commensal-derived primary and secondary bacterial metabolites emerged as potent modulators of hosts susceptibility to infection. Our results provide evidence that variations in SCFA production in mice fed fibre-rich chow-based diets modulate susceptibility to colonization with Enterobacteriaceae not only in antibiotic-disturbed ecosystems but even in undisturbed microbial communities. These findings emphasise the need for microbiota normalization across laboratory mouse lines for infection experiments with the model-pathogen C. rodentium independent of investigations of diet and antibiotic usage., Author summary The distinct composition of the gut microbiota in each individual results in variable metabolic activity and output of these communities, which influences the host, including resistance to enteric pathogens. Lack of reproducibility in biomedical research is nowadays frequently attributed to the microbiota, but little is known about which specific members and metabolites contribute to disease severity. Here, we use genetically identical mouse lines with variable microbiota compositions on a standardized diet and observed highly variable colonization with the enteric pathogen Citrobacter rodentium without antibiotics intervention. We found the same differences in formerly germ-free animals harbouring the respective donors microbiota and also in vitro by coculturing cecal bacteria from resistant and susceptible animals with C. rodentium showing that the phenotype is fully dependent on differences in the microbiota. We analysed the microbiome composition and found a higher abundance of butyrate-producing bacteria as well as increased levels of butyrate in resistant mice. By supplementation of susceptible and germ-free animals with butyrate, we could significantly lower the levels of colonization highlighting that commensal-derived primary and secondary bacterial metabolites are highly variable between laboratory animals from different vendors and are potent modulators of hosts susceptibility to infection with C. rodentium.

Summary Gut colonization with multidrug-resistant (MDR) bacteria enhances the risk of bloodstream infections in susceptible individuals. We demonstrate highly variable degrees of ex vivo colonization resistance against a carbapenem-resistant Klebsiella pneumoniae strain in human feces samples and subsequently isolate diverse K. oxytoca strains from protected donors. Several of these K. oxytoca strains reduce gut colonization of MDR K. pneumoniae strains in antibiotic-treated and gnotobiotic mouse models. Comparative analysis of K. oxytoca strains coupled with CRISPR-Cas9-mediated deletion of casA, a protein essential for utilization of selected beta-glucosides, identified competition for specific carbohydrates as key in promoting colonization resistance. In addition to direct competition between K. oxytoca and K. pneumoniae, cooperation with additional commensals is required to reestablish full colonization resistance and gut decolonization. Finally, humanized microbiota mice generated from K. pneumoniae-susceptible donors are protected by K. oxytoca administration, demonstrating the potential of commensal K. oxytoca strains as next-generation probiotics.

Summary Identifying distinctive taxa for micro-biome-related diseases is considered key to the establishment of diagnosis and therapy options in precision medicine and imposes high demands on the accuracy of micro-biome analysis techniques. We propose an alignment- and reference- free subsequence based 16S rRNA data analysis, as a new paradigm for micro-biome phenotype and biomarker detection. Our method, called DiTaxa, substitutes standard operational taxonomic unit (OTU)-clustering by segmenting 16S rRNA reads into the most frequent variable-length subsequences. We compared the performance of DiTaxa to the state-of-the-art methods in phenotype and biomarker detection, using human-associated 16S rRNA samples for periodontal disease, rheumatoid arthritis and inflammatory bowel diseases, as well as a synthetic benchmark dataset. DiTaxa performed competitively to the k-mer based state-of-the-art approach in phenotype prediction while outperforming the OTU-based state-of-the-art approach in finding biomarkers in both resolution and coverage evaluated over known links from literature and synthetic benchmark datasets. Availability and implementation DiTaxa is available under the Apache 2 license at http://llp.berkeley.edu/ditaxa. Supplementary information Supplementary data are available at Bioinformatics online.

Fasting can prolong survival and reduce disease burden in rodent models, and possibly in humans. The relationship between diet, gut microbiota, immune system and host (patho)physiology has only recently been explored, and information is lacking on how periodic fasting affects the gut microbiome in patients with metabolic syndrome (MetS). We show a 5-day fast (FAST) in humans, followed by a modified DASH diet is more effective than DASH alone (DASH) at reducing systolic blood pressure (SBP change measured by ABPM, 95% CI; FAST: [-7.053,-1.142], DASH: [-5.880,1.477]), need for antihypertensive medication (FAST: n=15 of n=35, DASH: n=6 of n=36 patients), and body-mass index at three months post intervention. Fasting altered the gut microbiome, impacting bacterial taxa and functional gene modules associated with the production of short-chain fatty acids (e.g. Faecalibacterium prausnitzii , Eubacterium rectale, Coprococcus comes ), previously linked to vascular health and immunity. Immunophenotyping and cross-system analyses revealed that SBP changes correlated with circulating Il-2 + TNFα + mucosa-associated invariant T (MAIT) cells (FDR-corr P(q) =0.044, Spearman’s rho=0.44), Il-17 - IFNγ + MAITs (FDR-corr P(q) =0.022, Spearman’s rho=0.49), and effector CD4 + T cells (FDR-corr P(q)=0.047, Spearman’s rho=0.43). By stratifying the fasting group into BP responders and non-responders, we identified a set of 76 microbial and 99 immune responder-specific features. Machine learning algorithms could predict long-term SBP responsiveness from baseline immunome data, identifying changes in effector CD8 + T cells, Th17 cells and Tregs as discriminators (Single-subject prediction: 71%). This is the first high-resolution multi-omics characterization of fasting in MetS. Fasting induced long-term reduction in body weight and SBP, accompanied by changes in microbiome and immune homeostasis. Our data implicate fasting as a promising non-pharmacological intervention in MetS.

ObjectivesRheumatoid arthritis (RA) has been associated with a relative expansion of faecal Prevotellaceae. To determine the microbiome composition and prevalence ofPrevotellaspp. in a group of individuals at increased risk for RA, but prior to the development of the disease.MethodsIn an ongoing cohort study of first-degree relatives (FDRs) of patients with RA, we identified ‘FDR controls’, asymptomatic and without autoantibodies, and individuals in pre-clinical RA stages, who had either developed anticitrullinated peptide antibodies or rheumatoid factor positivity and/or symptoms and signs associated with possible RA. Stool sampling and culture-independent microbiota analyses were performed followed by descriptive statistics and statistical analyses of community structures.ResultsA total of 133 participants were included, of which 50 were categorised as ‘FDR controls’ and 83 in ‘pre-clinical RA stages’. The microbiota of individuals in ‘pre-clinical RA stages’ was significantly altered compared with FDR controls. We found a significant enrichment of the bacterial family Prevotellaceae, particularlyPrevotellaspp., in the ‘pre-clinical RA’ group (p=0.04).ConclusionsPrevotellaspp. enrichment in individuals in pre-clinical stages of RA, before the onset of RA, suggests a role of intestinal dysbiosis in the development of RA.

The vast complexity of host-associated microbial ecosystems requires generation of host-specific gene catalogs to survey the functions and diversity of these communities. We generated a comprehensive resource, the integrated mouse gut metagenome catalog (iMGMC), comprising 4.6 million unique genes and 660 high-quality metagenome-assembled genomes (MAGs) linked to reconstructed full-length 16S rRNA gene sequences. iMGMC enables unprecedented coverage and taxonomic resolution, i.e. more than 89% of the identified taxa are not represented in any other databases. The tool (github.com/tillrobin/iMGMC) allowed characterizing the diversity and functions of prevalent and previously unknown microbial community members along the gastrointestinal tract. Moreover, we show that integration of MAGs and 16S rRNA gene data allows a more accurate prediction of functional profiles of communities than based on 16S rRNA amplicons alone. Integrated gene catalogs such as iMGMC are needed to enhance the resolution of numerous existing and future sequencing-based studies.

Background As the gut microbiota contributes to metabolic health, it is important to determine specific diet-microbiota interactions that influence host metabolism. Bile acids and dietary fat source can alter phenotypes of diet-induced obesity, but the interplay with intestinal microorganisms is unclear. Here, we investigated metabolic consequences of diets enriched in primary bile acids with or without addition of lard or palm oil, and studied gut microbiota structure and functions in mice. Results In combination with bile acids, dietary lard fed to male C57BL/6N mice for a period of 8 weeks enhanced fat mass accumulation in colonized, but not in germ-free mice when compared to palm oil. This was associated with impaired glucose tolerance, lower fasting insulin levels, lower counts of enteroendocrine cells, fatty liver, and elevated amounts of hepatic triglycerides, cholesteryl esters, and monounsaturated fatty acids. Lard- and bile acid-fed mice were characterized by shifts in dominant gut bacterial communities, including decreased relative abundances of Lachnospiraceae and increased occurrence of Desulfovibrionaceae and the species Clostridium lactatifermentans and Flintibacter butyricus. Metatranscriptomic analysis revealed shifts in microbial functions, including lipid and amino acid metabolism. Conclusions Caution is required when interpreting data from diet-induced obesity models due to varying effects of dietary fat source. Detrimental metabolic consequences of a diet enriched with lard and primary bile acids were dependent on microbial colonization of the host and were linked to hepatic lipid rearrangements and to alterations of dominant bacterial communities in the cecum. Electronic supplementary material The online version of this article (10.1186/s40168-018-0510-8) contains supplementary material, which is available to authorized users.

Background Prevotella spp. have been identified as highly enriched in the intestinal microbiota of patients newly diagnosed with rheumatoid arthritis (RA), suggesting a role in the development of the disease.1 Sequence homology between RA-specific autoantigens and proteins of Prevotella copri have been reported.2 However, the role of these bacteriae in the pathogenesis of the disease is not yet established. Objectives To determine the microbiome composition and prevalence of Prevotella spp. in different pre-clinical phases of RA, in a group of individuals at risk for RA, namely first degree relatives of patients with RA (RA-FDR). Methods In an ongoing cohort study of RA-FDR, enrolling individuals without clinical evidence of RA at inclusion, we categorised individuals in the following groups: ‘healthy controls’: asymptomatic RA-FDR without any autoantibodies or symptoms associated with possible RA; ‘pre-clinical RA’: individuals with ‘systemic autoimmunity associated with RA’ defined by the presence of anti-citrullinated peptide antibodies (ACPA) or rheumatoid factor (RF) and/or symptomatic individuals with clinically suspect arthralgias or unclassified arthritis. Participants provided stool samples for microbiome analysis. We excluded subjects who had undergone antibiotic therapy within the last 3 months, with known history of inflammatory bowel disease and/or gastrointestinal tract surgery ever. Stool samples processing and microbial diversity culture-independent analyses were performed. After DNA extraction, the V4 region of the 16S rRNA gene was amplified using barcoded primers and sequencing was done on an Illumina MiSeq. Statistical analyses of community structures were performed.3 Results Of the 134 participants enrolled, 51 were categorised as ‘healthy controls’ and 83 as ‘pre-clinical RA’. table 1 shows the general characteristics of the two groups. The microbiota of ‘pre-clinical RA’ individuals was significantly altered compared to ‘healthy controls’, with abundance of specific bacteria, particularly an enrichment of Prevotella spp. (figure 1). Conclusions Individuals at risk of RA who have developed systemic autoimmunity associated with RA and/or symptoms, have enrichment of Prevotella spp in comparison with healthy controls. Our findings support the hypothesis of a causal role of Prevotella spp in the development of RA, which could lead to future attempts to interfere with its intestinal colonisation during the preclinical stages of disease. References [1] Scher JU, et al. eLife2013;2:e01202. [2] Pianta A, et al. J Clin Invest2017;127(8):2946–56. [3] Segata N, et al. Genome Biology2011;12:R60. Disclosure of Interest None declared

Background Shotgun metagenome data sets of microbial communities are highly diverse, not only due to the natural variation of the underlying biological systems, but also due to differences in laboratory protocols, replicate numbers, and sequencing technologies. Accordingly, to effectively assess the performance of metagenomic analysis software, a wide range of benchmark data sets are required. Results We describe the CAMISIM microbial community and metagenome simulator. The software can model different microbial abundance profiles, multi-sample time series, and differential abundance studies, includes real and simulated strain-level diversity, and generates second- and third-generation sequencing data from taxonomic profiles or de novo. Gold standards are created for sequence assembly, genome binning, taxonomic binning, and taxonomic profiling. CAMSIM generated the benchmark data sets of the first CAMI challenge. For two simulated multi-sample data sets of the human and mouse gut microbiomes, we observed high functional congruence to the real data. As further applications, we investigated the effect of varying evolutionary genome divergence, sequencing depth, and read error profiles on two popular metagenome assemblers, MEGAHIT, and metaSPAdes, on several thousand small data sets generated with CAMISIM. Conclusions CAMISIM can simulate a wide variety of microbial communities and metagenome data sets together with standards of truth for method evaluation. All data sets and the software are freely available at https://github.com/CAMI-challenge/CAMISIM Electronic supplementary material The online version of this article (10.1186/s40168-019-0633-6) contains supplementary material, which is available to authorized users.

Summary Inflammatory bowel disease comprises a group of heterogeneous diseases characterized by chronic and relapsing mucosal inflammation. Alterations in microbiota composition have been proposed to contribute to disease development, but no uniform signatures have yet been identified. Here, we compare the ability of a diverse set of microbial communities to exacerbate intestinal inflammation after chemical damage to the intestinal barrier. Strikingly, genetically identical wild-type mice differing only in their microbiota composition varied strongly in their colitis susceptibility. Transfer of distinct colitogenic communities in gene-deficient mice revealed that they triggered disease via opposing pathways either independent or dependent on adaptive immunity, specifically requiring antigen-specific CD4+ T cells. Our data provide evidence for the concept that microbial communities may alter disease susceptibility via different immune pathways despite eventually resulting in similar host pathology. This suggests a potential benefit for personalizing IBD therapies according to patient-specific microbiota signatures., Graphical Abstract, Highlights • Gut microbiota composition modulates colitis severity in immunocompetent hosts • Colitogenic microbiota drive colitis via innate or adaptive immunity • Distinct microbiota members induce pathogenic CD4+ T cells to drive colitis, Alterations in the microbiota contribute to the development of intestinal inflammation. Roy et al. demonstrate that distinct intestinal microbial communities cause colitis via opposing effector mechanisms independent or dependent on adaptive immunity. Their findings suggest that personalized immunomodulatory treatment according to distinct microbial signatures may be beneficial for IBD patients.