Our understanding of how the gut microbiome interacts with its human host has been restrained by limited access to longitudinal datasets to examine stability and dynamics, and by having only a few isolates to test mechanistic hypotheses. Here, we present the Broad Institute-OpenBiome Microbiome Library (BIO-ML), a comprehensive collection of 7,758 gut bacterial isolates paired with 3,632 genome sequences and longitudinal multi-omics data. We show that microbial species maintain stable population sizes within and across humans and that commonly used ‘omics’ survey methods are more reliable when using averages over multiple days of sampling. Variation of gut metabolites within people over time is associated with amino acid levels, and differences across people are associated with differences in bile acids. Finally, we show that genomic diversification can be used to infer eco-evolutionary dynamics and in vivo selection pressures for strains within individuals. The BIO-ML is a unique resource designed to enable hypothesis-driven microbiome research.
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Sequencing and genomic data were deposited on the NCBI, under BioProject PRJNA544527.
BioSample accession numbers for isolate genome assemblies:
BioSample accession numbers for raw 16S data:
BioSample accession numbers for metagenomic data:
The processed metabolomics data is available at the NIH Common Fund’s Metabolomics Data Repository and Coordinating Center (supported by NIH grant, U01-DK097430) website, the Metabolomics Workbench, http://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000804.
Scripts and command lines used to analyze the sequencing and genomic data are available at https://github.com/almlab/BIO-ML.
The library of isolates is maintained and stored at the Broad Institute and strains will be made available for purchase upon request by researchers through a Broad Institute online platform: https://www.broadinstitute.org/bio-ml
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The authors are thankful to M. Sovie, C. Kim, W. Kelley, E. Lee, W. Pettee, J. Watson and P. Panchal from OpenBiome for their assistance in processing materials and donor metadata used in this study. This work was funded by a grant from the Broad Institute (Broad Next 10 grant 4000017).
M.S. and E.J.A. are co-founders and shareholders of Finch Therapeutics, a company that specializes in microbiome-targeted therapeutics.
Peer review information: Alison Farrell is the primary editor on this article and managed its editorial process and peer review in collaboration with the rest of the editorial team.
Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
a, 16S phylogenetic tree of the 7,758 isolates in the BIO-ML. Lineages are colored by phylum. b, Depiction of the distribution of 1,347 isolates across 24 bacterial species (y axis) over time (x axis) that were whole-genome sequenced. c, Depiction of the distribution of 1,168 samples across individuals (y axis) and over time (x axis) that were processed for 16S amplicon sequencing. d, Depiction of the distribution of 563 samples across individuals and over time that were processed for shotgun metagenomic sequencing. e, Depiction of the distribution of 179 samples across individuals and over time that were processed for metabolomic study.
a, Abundance-weighted taxonomic coverage of the library of bacterial isolates (7,758 isolates) (y axis), compared to the diversity observed through culture-independent 16S amplicon sequencing (x axis). Eleven donors were used to build the library of isolates. The phylogenetic diversity of isolates was measured with 16S sanger sequencing, and this was compared to the total diversity observed in the 16S sequence data obtained from 1,168 samples from 90 individual donors of the BIO-ML. Taxonomic coverage was evaluated using different 16S OTU clustering thresholds, from 90% to 100% (ASV) similarity. b, Phylogenomic tree of the 3,632 genomes of the BIO-ML. Branches are colored by phylum.
Extended Data Fig. 3 The library of genomes contain multiple species within the Faecalibacterium and Akkermansia genera.
Phylogenetic trees of Faecalibacterium (a) and Akkermansia (b) genomes were reconstructed using the concatenate alignment of ribosomal proteins (see Methods). We used RAxML to reconstruct the tree, using the PROTGAMMALGF substitution model. Pairwise Mash distances are represented on the right of each tree. Within each major clade, pairwise Mash distances were lower than 0.05, the threshold used to define species taxonomic units. Between clades, pairwise distances were higher than 0.05. Genomes in the F. prausnitzii and A. muciniphila clades have Mash distances with corresponding NCBI reference genomes that were lower than 0.05. Two different Akkermansia species are present in our genome library. At least two different Faecalibacterium species are present in the genome library.
Extended Data Fig. 4 Stability and conservation of microbiome species over time within and across people.
a, Non-metric multidimensional scaling (NMDS) plot showing 16S community structure (Bray–Curtis distances) across long-term time series from ten stool donors. Samples are colored by donors (right). Donors maintain unique microbial signatures over many months to years (ANOSIM, P < 0.0001). b, The black points show the median abundance comparisons, and the red points show the results for a single, randomly drawn sample. Species abundances are conserved across donor pairs. The spread in the red points is larger than for the black points, indicating the median abundances show a tighter correlation across donors (black points Pearson’s R2 = 0.25; red points Pearson’s R2 = 0.19).
Extended Data Fig. 5 Stability and conservation of microbiome functions over time within and across people.
a, NMDS plot showing functional structure (Bray–Curtis distances) across long-term time series from four stool donors. Donors maintain unique functional signatures over many months-to-years. b, COG abundances are conserved across donor pairs. The black points show the median abundance comparisons, and the red points show the results for a single, randomly drawn sample. The spread in the red points is larger than that for the black points, indicating the median abundances show a tighter correlation across donors (black points Pearson’s R2 = 0.88; red points Pearson’s R2 = 0.77).
Extended Data Fig. 6 Averaging taxa abundances across time points improves the identification of species–species correlations.
a, Correlation matrix of log median ASV relative abundances across ten donors with long, dense time series (that is cross-sectional correlations) filtered to only look at abundant SVs with average frequencies of ≥0.01 across the dataset. b, Distribution of correlation coefficients from panel a. Dashed lines show the significance threshold (P < 0.05). Correlations beyond this threshold were used to infer a cross-sectional correlation network from the full dataset. c, The fraction of edges from the cross-sectional correlation network inferred from the full dataset that are captured by random subsampling of donor time series. Choosing a single sample from each donor only captures ~40% of ‘true’ network edges (number of iterations = 10).
Extended Data Fig. 7 Metabolomics data capture crossdonor variation as well as within-donor variation through time.
a, PC scores plot of all 179 samples for which metabolomic data were generated. Samples colored in gray correspond to subjects for which metabolomics data had been generated for less than six time points. Arrows connecting samples reflect the chronological order in which samples were collected. b, Dendrogram for donors for which metabolomics data had been generated for more than six time points. Metabolomes are colored by subject, as in a. The first two letters indicate the donor ID.
Extended Data Fig. 8 Bacterial taxa–metabolites correlation network reveals strong functional associations in the human gut.
Significant correlations between bacterial taxa and metabolite abundances (|Spearman’s rho| > 0.7, P < 0.01) suggest a link between eating meat and bacterial community composition. Alistipes and Subdoligranulum are strongly associated with the bile acid taurocholate and its derivatives. Subdoligranulum is also associated with carnitine, which has been linked to eating meat. Other taxa are associated with acids and lipids common to the gut environment.
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