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UTwastewaterARG

Identification of ARGs and MGEs in wastewater metagenomes

This repository contains a complete pipeline for identification of antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs) in metagenomes. It was carried out on a dedicated server in the Brazelton Lab at the University of Utah.

The batch script "utwaste-pipeline-master.sh" uses an assembly-based approach, beginning with quality control of the original sequencing reads and ending with the calculation of sequencing coverage of each predicted ARG and MGE in each metagenome.

The "plots" folder includes R code for generating plots with the tables generated by the pipeline.

Dependencies

Various python tools written by the Brazelton lab, documented in the following repositories:

https://github.com/Brazelton-Lab/seq-qc

https://github.com/Brazelton-Lab/seq-annot

https://github.com/Brazelton-Lab/lab_scripts

Main software dependencies

bbduk https://jgi.doe.gov/data-and-tools/software-tools/bbtools/bb-tools-user-guide/bbduk-guide/

Megahit https://github.com/voutcn/megahit

Prodigal https://github.com/hyattpd/Prodigal

AMRFinderPlus https://github.com/ncbi/amr

CoverM https://github.com/wwood/CoverM

Bowtie2 https://github.com/BenLangmead/bowtie2

samtools https://github.com/samtools/samtools

diamond https://github.com/bbuchfink/diamond

mobileOG https://github.com/clb21565/mobileOG-db/tree/main

VFDB http://www.mgc.ac.cn/VFs/download.htm

Citations

The overall assembly-based approach, sequence coverage calculations, and the seq-qc and seq-annot packages were described in:

Thornton, CT, Tanner, W, VanDerslice, J, Brazelton, WJ (2020) Localized effect of treated wastewater effluent on the resistome of an urban watershed. GigaScience. 9 (11): giaa125, https://doi.org/10.1093/gigascience/giaa125

Brazelton WJ, McGonigle JM, Motamedi S, Pendleton HL, Twing KI, Miller BC, Lowe WJ, Hoffman AM, Prator CA, Chadwick GL, Anderson RE, Thomas E, Butterfield DA, Aquino KA, Früh-Green GL, Schrenk MO, Lang SQ. 2022. Metabolic strategies shared by basement residents of the Lost City hydrothermal field. Applied and Environmental Microbiology: e00929-22. DOI: 10.1128/aem.00929-22.

In addition, each of the tools listed above should be cited if you use them in your own work.

Acknowledgements

Brazelton lab tools were created by Christopher Thornton and Alex Hyer. Analyses of ARGs and MGEs were further developed by Robin McArdel.

This work was funded by the Centers for Disease Control through a SHEPheRD RFTOP award to Matthew Samore, Jennifer Weidhaas, and James VanDerslice (University of Utah). The point of contact for this repository is William Brazelton.

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