meteor 2.0.2

Meteor - A plateform for quantitative metagenomic profiling of complex ecosystems

Meteor

Introduction

Meteor is a plateform for quantitative metagenomics profiling of complex ecosystems. Meteor relies on genes catalogue to perform species-level taxonomic profiling, functional analysis and strain-level population structure inference.

Dependencies

Meteor requires:

• python>=3.10
• bowtie2
• bcftools
• FastTree
• ete3

Installation

Meteor is available with conda which includes all its dependencies:

conda create --name meteor -c conda-forge -c bioconda  -c aghozlane meteor

Or with pip with a recent Python>=3.10 from the cloned directory:

pip install meteor

You can test the installation of meteor with:

meteor test

Getting started

A basic usage of meteor will require to:

1. Download or build a reference catalogue
2. Structure the raw fastq files
3. Map reads against the reference catalogue
4. Compute taxonomical and/or functional abundances
5. Strain profiling

1. Download or build a custom reference

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Meteor requires to download locally a microbial gene catalogue specif, either in 'full' or 'light' version. The 'full' version contains all genes of the catalogue, whereas the 'light' version contains only the marker genes that will be used to infer species abundance profiles. Of note, no functional profiling can be performed when using the 'light' version of a catalogue.

Ten catalogues are currently available:

Microbial gene catalogue	<name>	Genes count (M)	Metagenomic Species Pan-genomes (MSPs)	Size (full) (GB)	Size (light) (GB)	Description
Felis catus	cat_gut	1.3	344	2.7	0.9	link
Gallus gallus domesticus	chicken_caecal	13.6	2420	22.8	4.8	link
Canis lupus familiaris	dog_gut	0.95	234	1.7	0.3	link
Homo sapiens gut	human_gut	10.4	1990	15.1	3.2	link
Homo sapiens oral	human_oral	8.4	853	16.4	3.2	link
Homo sapiens skin	human_skin	2.9	392	4.6	0.9	link
Mus musculus	mouse_gut	5.0	1252	10.3	3.4	link
Oryctolagus cuniculus	rabbit_gut	5.7	1053	11.1	3.5	link
Rattus norvegicus	rat_gut	5.9	1627	8.4	2.0	link
Sus domesticus	pig_gut	9.3	1523	8.4	378	link

These references can be downloaded with the following command:

meteor download -i <name> -c -o <refdir>

The 'light' catalogues are available with the tag (--fast) :

meteor download -i <name> -c --fast -o <refdir>

Users can also import custom gene catalogue with the command:

meteor build -i <fastafile> -n <name> -o <refdir> -t <threads>

2. Import fastq

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Meteor requires a first of fastq indexing:

meteor fastq -i <fastqdir>  [-p paired reads] -n <projectname> -o <outputdir>

When multiple sequencing are available for a library, the option -m allows to group these samples. Example:

Illumina_lib1-SAMPLE_01.fastq
Illumina_lib1-SAMPLE_02.fastq
Illumina_lib2-SAMPLE_01.fastq
Illumina_lib2-SAMPLE_02.fastq

In this case, the following command will group these samples the same library:

meteor fastq -i ./  -m SAMPLE_\\d+ -n projectname -o outputdir

3. Mapping

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The raw fastq files are mapped against a catalogue to generate a gene count table with the following command:

meteor mapping -i <fastqdir/sampledir> -r <refdir> -o <mappingdir>

We recommend to first filter out reads with low-quality, length < 60nt or belonging to the host.

4. Taxonomic and functional profiling

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Genes from the catalogue are clustered into Metagenomic Species Pangeomes (MSP) with MSPminer, and are functionnaly annotated against KEGG r107, DBcan (carbohydate active enzymes) and MUSTARD (antiobitic resistant determinants).

MSP and functional profiles are computed from the gene count table with the following command:

meteor profile -i <mappingdir> -o <profiledir> -r <refdir> -n coverage

The "-n" parameter ensures read count normalization for gene length. If omitted, no normalization will be performed on the gene table.

This profiling step will generate:

• species abundance table;
• ARD abundance table (full catalogue only);
• DBCAN abundance table (full catalogue only);
• Gut Metabolic Modules (GMM) abundance table (from the KO annotation) (full catalogue only).

5. Merging

To merge output from different samples into a single table, use the following command:

meteor merge -i <profiledir> -o <mergingdir> --fast

The '--fast' parameter prevent merging of the gene count tables, so that only species and functions table will be merged.

5. Strain profiling

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The METEOR team

The main contributors to METEOR:

• Franck Gauthier
• Amine Ghozlane
• Florian Plaza Oñate
• Nicolas Pons
• Florence Thirion

Acknowledgements

Special thanks to the following people:

• Mathieu Almeida
• Emmanuelle Le Chatelier