Improved eukaryotic detection compatible with large-scale automated analysis of metagenomes

Bazant, W., Blevins, A. S., Crouch, K. and Beiting, D. P. (2023) Improved eukaryotic detection compatible with large-scale automated analysis of metagenomes. Microbiome, 11, 72. (doi: 10.1186/s40168-023-01505-1) (PMID:37032329) (PMCID:PMC10084625)

Abstract

Background: Eukaryotes such as fungi and protists frequently accompany bacteria and archaea in microbial communities. Unfortunately, their presence is difficult to study with “shotgun” metagenomic sequencing since prokaryotic signals dominate in most environments. Recent methods for eukaryotic detection use eukaryote-specific marker genes, but they do not incorporate strategies to handle the presence of eukaryotes that are not represented in the reference marker gene set, and they are not compatible with web-based tools for downstream analysis. Results: Here, we present CORRAL (for Clustering Of Related Reference ALignments), a tool for the identification of eukaryotes in shotgun metagenomic data based on alignments to eukaryote-specific marker genes and Markov clustering. Using a combination of simulated datasets, mock community standards, and large publicly available human microbiome studies, we demonstrate that our method is not only sensitive and accurate but is also capable of inferring the presence of eukaryotes not included in the marker gene reference, such as novel strains. Finally, we deploy CORRAL on our MicrobiomeDB.org resource, producing an atlas of eukaryotes present in various environments of the human body and linking their presence to study covariates. Conclusions: CORRAL allows eukaryotic detection to be automated and carried out at scale. Implementation of CORRAL in MicrobiomeDB.org creates a running atlas of microbial eukaryotes in metagenomic studies. Since our approach is independent of the reference used, it may be applicable to other contexts where shotgun metagenomic reads are matched against redundant but non-exhaustive databases, such as the identification of bacterial virulence genes or taxonomic classification of viral reads.

• Item Type: Articles
• Additional Information: This work was partially supported by a grant from the Bill and Melinda Gates Foundation (D.P.B. and A.S.B.) and Astarte Medical (D.P.B. and W.B.).
• Keywords: Software, metagenome, shotgun metagenomics, microbial eukaryotes, bioinformatics, fungi, mycobiome.
• Status: Published
• Refereed: Yes
• Glasgow Author(s) Enlighten ID: Bazant, Mr Wojtek and Crouch, Dr Kathryn
• Authors: Bazant, W., Blevins, A. S., Crouch, K., and Beiting, D. P.
• College/School: College of Medical Veterinary and Life Sciences > School of Infection & Immunity
• Journal Name: Microbiome
• Publisher: BioMed Central
• ISSN: 2049-2618
• ISSN (Online): 2049-2618
• Copyright Holders: Copyright © 2023 The Authors
• First Published: First published in Microbiome 11: 72
• Publisher Policy: Reproduced under a Creative Commons License