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Microbial analysis using Galaxy

Community, Projects, Workflows, Tools, and Training

By Paul Zierep (Galaxy Freiburg)

OSA2Micro, Turin, July 2023

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Content

  • Galaxy
    • Introduction
    • Frontend
    • Running a tool
    • Interactive tools
    • Workflows
    • Architecture
  • The EU server
  • Training and the Galaxy Training Network
  • microGalaxy Community
    • Community
    • Survey
  • Galaxy for Genomics and Metatranscriptomics
    • Tools
    • Workflows
  • Microbiome analysis
    • Tools
    • Workflows

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Galaxy Introduction - Huge increase in Sequencing Data

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Galaxy Introduction

  • Strong increase of generating scientific data: Sequencing technology has become fast and costs are low
  • Galaxy allows bioinformatics analysis for everyone
  • No programming skills required
  • Enable wet-lab scientists to do their own data analysis
  • Open source framework for high-throughput sequencing data and other big data analysis

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Galaxy Introduction

  • Galaxy is a gateway for transparent & reproducible data analysis using FAIR principles
  • Easy accessing (no installation) & sharing of data, tools, analysis, workflows
  • Multiple interfaces
    • intuitive web portals for biologists
    • unified API for bioinformaticians
  • International: 138+ public instances, 12,000+ citations
  • UseGalaxy.eu, UseGalaxy.org.au, UseGalaxy.org ...
  • Funded as international infrastructure project (EuroScienceGateway, ELIXIR, EOSC)

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Galaxy Introduction

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Galaxy GUI

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Galaxy Tools

  • Galaxy tools are wrappers that export inputs and outputs of command-line tools to the Galaxy GUI
  • > 9.000 tools
  • > 3.000 tools on usegalaxy.eu
  • Large tool suits (Mothur > 100)
  • New tools can be added by anyone via an open infrastructure
  • Internally tools require a conda, docker and/or singularity container

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Galaxy behind the scenes - Wrapping a tool

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Wrapping a tool

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Update infrastructure

Planemo: a command-line toolkit for developing, deploying, and executing scientific data analyses, 2022

Training

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Running a tool

Select your

  • Input data from history
  • Parameter for analysis
  • Output data

Each tool comes with

  • Documentation
  • Example input and results
  • Reference
  • Versioning

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Running a tool - database interaction

  • Admin based data management via Data Managers
  • No limit for DBs
  • No user quota increase

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Running a tool - interactive tools

  • RStudio
  • Jupyter Notebooks and specific flavors
    • Interactive MGnify Notebook
    • ML
    • Material Science (PyIron)
  • Interactive Hdfview tool
  • Pavian
  • Phinch

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Running a tool - interactive tools (Phinch)

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Running a tool - interactive tools (Phinch)

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Running a tool - interactive tools (Phinch)

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History - handle the data

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History - handle the data

  • Move data between histories without quota (or actual space) increase
  • No limit of histories
  • Can be shared, downloaded, published

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Data Preview and visualization

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Data Preview and visualization

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Data Preview and visualization

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Workflow building

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Workflow building

How?

  • Extracted from history (step-by-step)
  • Built manually (graphical workflow editor)
  • Import a shared workflow

Why?

  • Automatize your analysis
  • Re-run same analysis on different inputs or reproduce results
  • Change parameter, tool version, DB, workflows logic (add, remove, modify)
  • Use/create sub workflows
  • Share them, publish them (peer-reviewed in IWC!) + WorkflowHub

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Workflow building - graphical workflow editor

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Workflow building - share, import, store, update

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The European Galaxy server

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Galaxy - Training - Gallantries

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Galaxy - Training

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Galaxy - Training

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Galaxy - Training

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Galaxy - Training

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Galaxy - learn more

How?

  • Join training (https://galaxyproject.org/events/) e.g. GTN Smörgåsbord 2024
  • Galaxy Training Network https://training.galaxyproject.org/
  • Matrix/Element chat: https://matrix.to/#/#galaxyproject_Lobby:gitter.im
  • Try it out (free usage of the servers with 250 GB space)

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microGalaxy Community - a community of practice for everything related to micro*

  • https://galaxyproject.org/projects/microbial/
  • https://microgalaxy.usegalaxy.eu/
  • Roadmap development
  • Collect tools, workflows and tutorials
  • Bring people together and support each other
  • https://lists.galaxyproject.org/lists/microgalaxy.lists.galaxyproject.org/
  • Next community call: during GCC

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microGalaxy Community - Working Groups

  • Tool working groups
    • Community oriented updated tool list
      • Increasing the findability, visibility, and impact�of Galaxy tools for specialized scientific Communities�(https://biohackathon-europe.org/ - Project)
    • Coordinate:
      • New wrapper development
      • Updates
      • Pitfalls (Huge tools suits)
      • Discuss ideas

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microGalaxy Community - a community of practice for everything related to micro*

  • Community oriented paper - working groups
    • Survey
      • What is the need, what are the problems �for the community ?
    • Advertisement
      • Target the global south !
    • Aggregate information
    • Use cases
      • Learn from others
    • Roadmap
      • Next 2, 5, 10 years

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microGalaxy Community - survey

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microGalaxy Community - survey

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microGalaxy Community - survey

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microGalaxy Community - survey

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microGalaxy Community - survey

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microGalaxy Community - survey

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microGalaxy Community - survey

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microGalaxy Community - survey

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microGalaxy Community - survey

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microGalaxy Community - survey

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microGalaxy Community - survey

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Galaxy for microbial analysis

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Galaxy for genomics and transcriptomics

  • Tools
    • Preprocessing
      • Long reads (Oxford nanopore), short reads (Illumina)
    • Assembly
      • Flye (de novo - long reads), MEGAHIT (de novo), SPAdes (de novo), Quast (evaluation tool)

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Galaxy for genomics and transcriptomics

  • Tools
    • Gene prediction
      • Prodigal
    • Gene annotation
      • Prokka, Bakta, BLAST
    • Functional annotation
      • eggNOG, InterProScan, Diamond

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Galaxy for genomics and transcriptomics

  • Tools
    • Gene prediction
      • Prodigal
    • Gene annotation
      • Prokka, Bakta, BLAST
    • Functional annotation
      • eggNOG, InterProScan, Diamond

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Galaxy for genomics and transcriptomics

  • Tools
    • Mapping
      • STAR, HISAT2, BBtools, Bowtie2, Burrows-Wheeler Aligner (BWA - mapping)

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Galaxy for genomics and transcriptomics

  • Tools
    • Gene cluster detection
      • antiSMASH
    • Pangenome building
      • Scoary
    • Visulisation
      • JBrowse

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Galaxy for genomics and transcriptomics

  • Workflows and Trainings
    • Assembly
    • Genome Annotation
    • Transcriptomics
    • Variant Analysis
      • M. tuberculosis
      • SARS-CoV-2

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Galaxy for microbiome (meta*omics) analysis

  • Tools
    • Phylogenetic profiling
      • Amplicon based
        • Dada2, Mothur, ITSx
      • Read based
        • Kraken2, MetaPhlAn
    • Functional profiling
      • HUMAnN
    • Assembly
      • metaSpades, metaQuast
    • Binning
      • Das Tool, MetaBAT2, SemiBin, CONCOCT
    • Comparative genomics
      • DeSeq2
    • Predictive Metagenomics
      • DeepMicro

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Galaxy for microbiome analysis

  • Workflows and Training
    • GTN (Metagenomics: 13)
    • Amplicon based analysis
    • Read-based analysis
    • (foodborne)-Pathogens detection and tracking
    • ML base H. pylori detection and classification

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Amplicon

analysis

https://astrobiomike.github.io/

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Amplicon analysis

Comparison: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6964864/

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Amplicon analysis - BeerDeCoded - Street Science

BeerDeCoded: the open beer metagenome project, F1000Res, 2017

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Amplicon analysis - BeerDeCoded

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https://astrobiomike.github.io/

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ASaiM-(MT): a Galaxy-based framework to analyze microbiome data

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ASaiM-(MT): a Galaxy-based framework to analyze microbiome data

  • Read-based analysis
  • MetaPhlAn - uses clade-specific markers to identify and quantify the relative abundance of microbial taxa present in the sample
  • HUMAnN - compare functional pathways across different samples.

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Accessible and scalable

pipelines for fast and easy

(foodborne) pathogens

detection and tracking

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Predictive Metagenomics - Identification of parameters indicative of disease development by Helicobacter pylori predict.

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Identification of parameters indicative of disease development by Helicobacter pylori predict.

  • Input
    • microbiome composition (16S rDNA)
      • diversity and taxonomic composition
    • microbiome activity
      • RNA-sequencing
        • microbial gene expression
      • metabolite sampling (MS)
    • immune cell activity (‘immune status’)
      • leukocyte RNA-sequencing
    • H. pylori whole-genome sequencing (WGS)
      • H. pylori genotype in the infection group
    • + public data to enrich the data

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Identification of parameters indicative of disease development by Helicobacter pylori predict

16S rDNA

gene expression

metabolites

immune status

H. pylori SNPs

0.1,0.3,0.6

0.2,0.4,0.4

Infected / Not-infected

H. pylori benign/

malign

0

-

1

0

1

1

0

-

ML

WF1

WF2

WF3

WF4

WF5

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Curse of dimensionality

  • Rule of thumb: 5 training examples for each dimension in the representation.
  • DeepMicro: Transform high-dimensional microbiome data into a robust low-dimensional representation

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Curse of dimensionality - DeepMicro

DeepMicro: deep representation learning for disease prediction based

on microbiome data, 2020, Scientific Reports

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Thank you very much for your attention !

  • Try Galaxy for yourself !
  • We’re open for project ideas and collaborations.
  • Questions ?