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Abstract

The oesophageal microbiome is thought to contribute to the pathogenesis of oesophageal cancer. However, investigations using culture and molecular barcodes have provided only a low-resolution view of this important microbial community. We therefore explored the potential of culturomics and metagenomic binning to generate a catalogue of reference genomes from the healthy human oesophageal microbiome, alongside a comparison set from saliva.

Twenty-two distinct colonial morphotypes from healthy oesophageal samples were genome-sequenced. These fell into twelve species clusters, eleven of which represented previously defined species. Two isolates belonged to a novel species, which we have named . We performed metagenomic binning of reads generated from UK samples from this study alongside reads generated from Australian samples in a recent study. Metagenomic binning generated 136 medium or high-quality metagenome-assembled genomes (MAGs). MAGs were assigned to 56 species clusters, eight representing novel species which we have named . Granulicatella gullae, . Streptococcus gullae, . Nanosynbacter quadramensis, . Nanosynbacter gullae, . Nanosynbacter colneyensis, . Nanosynbacter norwichensis, . Nanosynococcus oralis and . Haemophilus gullae. Five of these novel species belong to the recently described phylum . Although members of the are known to inhabit the oral cavity, this is the first report of their presence in the oesophagus. Eighteen of the metagenomic species were, until recently, identified only by hard-to-remember alphanumeric placeholder designations. Here we illustrate the utility of a set of recently published arbitrary Latinate species names in providing user-friendly taxonomic labels for microbiome analyses.

Our non-redundant species catalogue contained 63 species derived from cultured isolates or MAGs. Mapping revealed that these species account for around half of the sequences in the oesophageal and saliva metagenomes. Although no species was present in all oesophageal samples, 60 species occurred in at least one oesophageal metagenome from either study, with 50 identified in both cohorts.

Recovery of genomes and discovery of new species represents an important step forward in our understanding of the oesophageal microbiome. The genes and genomes that we have released into the public domain will provide a base line for future comparative, mechanistic and intervention studies.

Funding
This study was supported by the:
  • Medical Research Council (Award MR/L015080/1)
    • Principal Award Recipient: MarkJ. Pallen
  • Biotechnology and Biological Sciences Research Council (Award BB/R012504/1)
    • Principal Award Recipient: MarkJ. Pallen
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
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/content/journal/acmi/10.1099/acmi.0.000558.v3
2023-06-26
2026-03-13

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References

  1. Gilroy R. Mags and cultured isolate Genomes Figshare 2022 [View Article]
    [Google Scholar]
  2. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I et al. Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 2021; 71:209–249 [View Article] [PubMed]
    [Google Scholar]
  3. Corning B, Copland AP, Frye JW. The esophageal microbiome in health and disease. Curr Gastroenterol Rep 2018; 20:39 [View Article] [PubMed]
    [Google Scholar]
  4. Kumar B, Lam S, Adam M, Gilroy R, Pallen MJ. The oesophageal microbiome and cancer: hope or hype?. Trends Microbiol 2022; 30:322–329 [View Article] [PubMed]
    [Google Scholar]
  5. May M, Abrams JA. Emerging insights into the esophageal Microbiome. Curr Treat Options Gastroenterol 2018; 16:72–85 [View Article] [PubMed]
    [Google Scholar]
  6. Bilen M. Strategies and advancements in human microbiome description and the importance of culturomics. Microb Pathog 2020; 149:104460 [View Article] [PubMed]
    [Google Scholar]
  7. Deshpande NP, Riordan SM, Castaño-Rodríguez N, Wilkins MR, Kaakoush NO. Signatures within the esophageal microbiome are associated with host genetics, age, and disease. Microbiome 2018; 6:227 [View Article] [PubMed]
    [Google Scholar]
  8. Gonzalez A, Vázquez-Baeza Y, Pettengill JB, Ottesen A, McDonald D et al. Avoiding pandemic fears in the subway and conquering the platypus. mSystems 2016; 1:e00050-16 [View Article] [PubMed]
    [Google Scholar]
  9. Kim CY, Lee M, Yang S, Kim K, Yong D et al. Human reference gut microbiome catalog including newly assembled genomes from under-represented Asian metagenomes. Genome Med 2021; 13:134 [View Article] [PubMed]
    [Google Scholar]
  10. Saheb Kashaf S, Proctor DM, Deming C, Saary P, Hölzer M et al. Integrating cultivation and metagenomics for a multi-kingdom view of skin microbiome diversity and functions. Nat Microbiol 2022; 7:169–179 [View Article] [PubMed]
    [Google Scholar]
  11. Gall A, Fero J, McCoy C, Claywell BC, Sanchez CA et al. Bacterial composition of the human upper gastrointestinal tract microbiome is dynamic and associated with genomic instability in a Barrett’s Esophagus Cohort. PLoS One 2015; 10:e0129055 [View Article] [PubMed]
    [Google Scholar]
  12. Gilroy R, Ravi A, Getino M, Pursley I, Horton DL et al. Extensive microbial diversity within the chicken gut microbiome revealed by metagenomics and culture. PeerJ 2021; 9:e10941 [View Article] [PubMed]
    [Google Scholar]
  13. Chen S, Zhou Y, Chen Y, Gu J. fastp: an ultra-fast all-in-one FASTQ preprocessor. Bioinformatics 2018; 34:i884–i890 [View Article] [PubMed]
    [Google Scholar]
  14. Bankevich A, Nurk S, Antipov D, Gurevich AA, Dvorkin M et al. SPAdes: a new genome assembly algorithm and its applications to single-cell sequencing. J Comput Biol 2012; 19:455–477 [View Article] [PubMed]
    [Google Scholar]
  15. Parks DH, Imelfort M, Skennerton CT, Hugenholtz P, Tyson GW. CheckM: assessing the quality of microbial genomes recovered from isolates, single cells, and metagenomes. Genome Res 2015; 25:1043–1055 [View Article] [PubMed]
    [Google Scholar]
  16. Olm MR, Brown CT, Brooks B, Banfield JF. dRep: a tool for fast and accurate genomic comparisons that enables improved genome recovery from metagenomes through de-replication. ISME J 2017; 11:2864–2868 [View Article] [PubMed]
    [Google Scholar]
  17. Chaumeil P-A, Mussig AJ, Hugenholtz P, Parks DH. GTDB-Tk: a toolkit to classify genomes with the genome taxonomy database. Bioinformatics 2019; 36:1925–1927 [View Article] [PubMed]
    [Google Scholar]
  18. Schwengers O, Hain T, Chakraborty T, Goesmann A. n.d. ReferenceSeeker: rapid determination of appropriate reference genomes. BioRxiv2019863621 [View Article]
    [Google Scholar]
  19. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ. Basic local alignment search tool. J Mol Biol 1990; 215:403–410 [View Article] [PubMed]
    [Google Scholar]
  20. Jain C, Rodriguez-R LM, Phillippy AM, Konstantinidis KT, Aluru S. High throughput ANI analysis of 90K prokaryotic genomes reveals clear species boundaries. Nat Commun 2018; 9:5114 [View Article] [PubMed]
    [Google Scholar]
  21. Ravi A, Halstead FD, Bamford A, Casey A, Thomson NM et al. Loss of microbial diversity and pathogen domination of the gut microbiota in critically ill patients. Microb Genom 2019; 5:e000293 [View Article] [PubMed]
    [Google Scholar]
  22. Connor TR, Loman NJ, Thompson S, Smith A, Southgate J et al. CLIMB (the Cloud Infrastructure for Microbial Bioinformatics): an online resource for the medical microbiology community. Microb Genom 2016; 2:e000086 [View Article] [PubMed]
    [Google Scholar]
  23. Langmead B, Salzberg SL. Fast gapped-read alignment with Bowtie 2. Nat Methods 2012; 9:357–359 [View Article] [PubMed]
    [Google Scholar]
  24. Eren AM, Esen ÖC, Quince C, Vineis JH, Morrison HG et al. Anvi’o: an advanced analysis and visualization platform for omics data. PeerJ 2015; 3:e1319 [View Article] [PubMed]
    [Google Scholar]
  25. Li H, Handsaker B, Wysoker A, Fennell T, Ruan J et al. The sequence alignment/map format and SAMtools. Bioinformatics 2009; 25:2078–2079 [View Article] [PubMed]
    [Google Scholar]
  26. Wu Y-W, Simmons BA, Singer SW. MaxBin 2.0: an automated binning algorithm to recover genomes from multiple metagenomic datasets. Bioinformatics 2016; 32:605–607 [View Article] [PubMed]
    [Google Scholar]
  27. Kang DD, Li F, Kirton E, Thomas A, Egan R et al. MetaBAT 2: an adaptive binning algorithm for robust and efficient genome reconstruction from metagenome assemblies. PeerJ 2019; 7:e7359 [View Article] [PubMed]
    [Google Scholar]
  28. Alneberg J, Bjarnason BS, de Bruijn I, Schirmer M, Quick J et al. Binning metagenomic contigs by coverage and composition. Nat Methods 2014; 11:1144–1146 [View Article] [PubMed]
    [Google Scholar]
  29. Sieber CMK, Probst AJ, Sharrar A, Thomas BC, Hess M et al. Recovery of genomes from metagenomes via a dereplication, aggregation and scoring strategy. Nat Microbiol 2018; 3:836–843 [View Article] [PubMed]
    [Google Scholar]
  30. Parks DH, Rinke C, Chuvochina M, Chaumeil P-A, Woodcroft BJ et al. Recovery of nearly 8,000 metagenome-assembled genomes substantially expands the tree of life. Nat Microbiol 2017; 2:1533–1542 [View Article] [PubMed]
    [Google Scholar]
  31. Brown CT, Hug LA, Thomas BC, Sharon I, Castelle CJ et al. Unusual biology across a group comprising more than 15% of domain Bacteria. Nature 2015; 523:208–211 [View Article] [PubMed]
    [Google Scholar]
  32. Pallen MJ, Rodriguez-R LM, Alikhan NF. Naming the unnamed: over 65,000 candidatus names for unnamed archaea and bacteria in the genome taxonomy database. Int J Syst Evol Microbiol 2022; 72: [View Article]
    [Google Scholar]
  33. Wickham H. ggplot2. WIREs Comp Stat 2011; 3:180–185 [View Article]
    [Google Scholar]
  34. Hyatt D, Chen GL, Locascio PF, Land ML, Larimer FW et al. Prodigal: prokaryotic gene recognition and translation initiation site identification. BMC Bioinformatics 2010; 11:119 [View Article] [PubMed]
    [Google Scholar]
  35. Segata N, Börnigen D, Morgan XC, Huttenhower C. PhyloPhlAn is a new method for improved phylogenetic and taxonomic placement of microbes. Nat Commun 2013; 4:2304 [View Article] [PubMed]
    [Google Scholar]
  36. Katoh K, Misawa K, Kuma K, Miyata T. MAFFT: a novel method for rapid multiple sequence alignment based on fast Fourier transform. Nucleic Acids Res 2002; 30:3059–3066 [View Article] [PubMed]
    [Google Scholar]
  37. Capella-Gutiérrez S, Silla-Martínez JM, Gabaldón T. trimAl: a tool for automated alignment trimming in large-scale phylogenetic analyses. Bioinformatics 2009; 25:1972–1973 [View Article] [PubMed]
    [Google Scholar]
  38. Darling AE, Mau B, Perna NT, Stajich JE. progressiveMauve: multiple genome alignment with gene gain, loss and rearrangement. PLoS One 2010; 5:e11147 [View Article] [PubMed]
    [Google Scholar]
  39. Hug LA, Baker BJ, Anantharaman K et al. A new view of the tree of life. Nat Microbiol 2016; 1:16048 [View Article] [PubMed]
    [Google Scholar]
  40. Edgar RC. MUSCLE: a multiple sequence alignment method with reduced time and space complexity. BMC Bioinformatics 2004; 113: [View Article] [PubMed]
    [Google Scholar]
  41. Price MN, Dehal PS, Arkin AP. FastTree 2--approximately maximum-likelihood trees for large alignments. PLoS One 2010; 5:e9490 [View Article] [PubMed]
    [Google Scholar]
  42. Letunic I, Bork P. Interactive Tree Of Life (iTOL) v5: an online tool for phylogenetic tree display and annotation. Nucleic Acids Res 2021; 49:W293–W296 [View Article] [PubMed]
    [Google Scholar]
  43. Shaiber A, Willis AD, Delmont TO, Roux S, Chen L-X et al. Functional and genetic markers of niche partitioning among enigmatic members of the human oral microbiome. Genome Biol 2020; 21:292 [View Article] [PubMed]
    [Google Scholar]
  44. Oksanen J, Kindt R, Legendre P. The vegan package. Commun Ecol Pack 2007; 10:719 [View Article]
    [Google Scholar]
  45. McMurdie PJ, Holmes S. phyloseq: an R package for reproducible interactive analysis and graphics of microbiome census data. PLoS One 2013; 8:e61217 [View Article] [PubMed]
    [Google Scholar]
  46. Castelle CJ, Banfield JF. Major new microbial groups expand diversity and alter our understanding of the tree of life. Cell 2018; 172:1181–1197 [View Article] [PubMed]
    [Google Scholar]
  47. Macfarlane S, Furrie E, Macfarlane GT, Dillon JF. Microbial colonization of the upper gastrointestinal tract in patients with Barrett’s esophagus. Clin Infect Dis 2007; 45:29–30 [View Article] [PubMed]
    [Google Scholar]
  48. Yamamura K, Baba Y, Nakagawa S et al. Human microbiome fusobacterium nucleatum in esophageal cancer tissue is associated with prognosis. Clin Cancer Res 2016; 22:5574–5581 [View Article] [PubMed]
    [Google Scholar]
  49. Pei Z, Bini EJ, Yang L, Zhou M, Francois F et al. Bacterial biota in the human distal esophagus. Proc Natl Acad Sci 2004; 101:4250–4255 [View Article] [PubMed]
    [Google Scholar]
  50. Elliott DRF, Walker AW, O’Donovan M, Parkhill J, Fitzgerald RC. A non-endoscopic device to sample the oesophageal microbiota: A case-control study. Lancet Gastroenterol Hepatol 2017; 2:32–42 [View Article] [PubMed]
    [Google Scholar]
  51. Lopetuso LR, Severgnini M, Pecere S et al. Esophageal microbiome signature in patients with Barrett’s esophagus and esophageal adenocarcinoma. PLoS One 2020; 15:e0231789 [View Article] [PubMed]
    [Google Scholar]
  52. Norder Grusell E, Dahlén G, Ruth M, Ny L, Quiding-Järbrink M et al. Bacterial flora of the human oral cavity, and the upper and lower esophagus. Dis Esophagus 2013; 26:84–90 [View Article] [PubMed]
    [Google Scholar]
  53. Okereke IC, Miller AL, Jupiter DC, Hamilton CF, Reep GL et al. Microbiota detection patterns correlate with presence and severity of Barrett’s esophagus. Front Cell Infect Microbiol 2021; 11:555072 [View Article] [PubMed]
    [Google Scholar]
  54. Parks DH, Chuvochina M, Rinke C, Mussig AJ, Chaumeil P-A et al. GTDB: an ongoing census of bacterial and archaeal diversity through a phylogenetically consistent, rank normalized and complete genome-based taxonomy. Nucleic Acids Res 2021; 50:D785–D794 [View Article] [PubMed]
    [Google Scholar]
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