From e04ff1e70dc2a7f3b68cdfc602de32be2e5c0d57 Mon Sep 17 00:00:00 2001 From: "Vitor C. Piro" Date: Wed, 7 Feb 2024 10:34:56 +0100 Subject: [PATCH 1/5] ganon v2.1.0 --- CMakeLists.txt | 2 +- LICENSE | 2 +- setup.py | 2 +- src/ganon/config.py | 2 +- 4 files changed, 4 insertions(+), 4 deletions(-) diff --git a/CMakeLists.txt b/CMakeLists.txt index 28c6f447..b2e4586d 100755 --- a/CMakeLists.txt +++ b/CMakeLists.txt @@ -3,7 +3,7 @@ # ============================================================================= cmake_minimum_required( VERSION 3.4 FATAL_ERROR ) -project( ganon VERSION 2.0.1 LANGUAGES CXX ) +project( ganon VERSION 2.1.0 LANGUAGES CXX ) # ----------------------------------------------------------------------------- # build setup diff --git a/LICENSE b/LICENSE index db103125..9f2a2e1a 100644 --- a/LICENSE +++ b/LICENSE @@ -1,6 +1,6 @@ MIT License -Copyright (c) 2023 Vitor C. Piro pirovc.github.io +Copyright (c) 2024 Vitor C. Piro pirovc.github.io Permission is hereby granted, free of charge, to any person obtaining a copy of this software and associated documentation files (the "Software"), to deal diff --git a/setup.py b/setup.py index e91c05a4..793492f8 100755 --- a/setup.py +++ b/setup.py @@ -13,7 +13,7 @@ def read(filename): setup( name="ganon", - version="2.0.1", + version="2.1.0", url="https://www.github.com/pirovc/ganon", license='MIT', author="Vitor C. Piro", diff --git a/src/ganon/config.py b/src/ganon/config.py index d2601c02..fd7514d5 100644 --- a/src/ganon/config.py +++ b/src/ganon/config.py @@ -8,7 +8,7 @@ class Config: - version = "2.0.1" + version = "2.1.0" path_exec = {"build": "", "classify": "", "get_seq_info": "", "genome_updater": ""} empty = False From d7046a989fc42de88ff4333c0621cd0b54c90a95 Mon Sep 17 00:00:00 2001 From: pirovc <4673375+pirovc@users.noreply.github.com> Date: Fri, 9 Feb 2024 15:39:27 +0100 Subject: [PATCH 2/5] Feature/input target sequence hibf (#283) * split sequences in files for --input-target sequence * sequence mode, tests, chunk size parsing acc2txid * fix docs * docs * Update tax_util.py * Update utils.py --- .travis.yml | 2 +- docs/classification.md | 6 +- docs/custom_databases.md | 179 ++++--- docs/default_databases.md | 19 +- docs/index.md | 26 +- docs/reports.md | 2 +- src/ganon-build/CommandLineParser.cpp | 2 +- src/ganon-build/GanonBuild.cpp | 132 ++--- src/ganon/build_update.py | 90 ++-- src/ganon/config.py | 14 +- src/ganon/tax_util.py | 106 ++-- src/ganon/util.py | 8 +- tests/aux/Aux.hpp | 9 +- tests/ganon-build/GanonBuild.test.cpp | 38 +- tests/ganon-build/data/SEQ0.fna.gz | Bin 0 -> 12852 bytes tests/ganon-build/data/SEQ1.fna.gz | Bin 0 -> 12247 bytes tests/ganon-build/data/SEQ10.fna.gz | Bin 0 -> 8395 bytes tests/ganon-build/data/SEQ11.fna.gz | Bin 0 -> 11653 bytes tests/ganon-build/data/SEQ12.fna.gz | Bin 0 -> 11834 bytes tests/ganon-build/data/SEQ13.fna.gz | Bin 0 -> 12031 bytes tests/ganon-build/data/SEQ14.fna.gz | Bin 0 -> 12164 bytes tests/ganon-build/data/SEQ15.fna.gz | Bin 0 -> 13009 bytes tests/ganon-build/data/SEQ16.fna.gz | Bin 0 -> 10645 bytes tests/ganon-build/data/SEQ17.fna.gz | Bin 0 -> 9094 bytes tests/ganon-build/data/SEQ18.fna.gz | Bin 0 -> 8577 bytes tests/ganon-build/data/SEQ19.fna.gz | Bin 0 -> 13164 bytes tests/ganon-build/data/SEQ2.fna.gz | Bin 0 -> 11952 bytes tests/ganon-build/data/SEQ20.fna.gz | Bin 0 -> 13090 bytes tests/ganon-build/data/SEQ21.fna.gz | Bin 0 -> 10347 bytes tests/ganon-build/data/SEQ22.fna.gz | Bin 0 -> 9746 bytes tests/ganon-build/data/SEQ23.fna.gz | Bin 0 -> 8020 bytes tests/ganon-build/data/SEQ24.fna.gz | Bin 0 -> 12401 bytes tests/ganon-build/data/SEQ3.fna.gz | Bin 0 -> 14053 bytes tests/ganon-build/data/SEQ4.fna.gz | Bin 0 -> 11840 bytes tests/ganon-build/data/SEQ5.fna.gz | Bin 0 -> 11567 bytes tests/ganon-build/data/SEQ6.fna.gz | Bin 0 -> 9159 bytes tests/ganon-build/data/SEQ7.fna.gz | Bin 0 -> 10158 bytes tests/ganon-build/data/SEQ8.fna.gz | Bin 0 -> 13415 bytes tests/ganon-build/data/SEQ9.fna.gz | Bin 0 -> 13300 bytes tests/ganon-build/data/mode.fasta.gz | Bin 275510 -> 0 bytes tests/ganon-build/data/mode_input.tsv | 50 +- tests/ganon/integration/test_build_custom.py | 492 ++++++++++++++++++- tests/ganon/utils.py | 148 ++++-- 43 files changed, 916 insertions(+), 407 deletions(-) create mode 100644 tests/ganon-build/data/SEQ0.fna.gz create mode 100644 tests/ganon-build/data/SEQ1.fna.gz create mode 100644 tests/ganon-build/data/SEQ10.fna.gz create mode 100644 tests/ganon-build/data/SEQ11.fna.gz create mode 100644 tests/ganon-build/data/SEQ12.fna.gz create mode 100644 tests/ganon-build/data/SEQ13.fna.gz create mode 100644 tests/ganon-build/data/SEQ14.fna.gz create mode 100644 tests/ganon-build/data/SEQ15.fna.gz create mode 100644 tests/ganon-build/data/SEQ16.fna.gz create mode 100644 tests/ganon-build/data/SEQ17.fna.gz create mode 100644 tests/ganon-build/data/SEQ18.fna.gz create mode 100644 tests/ganon-build/data/SEQ19.fna.gz create mode 100644 tests/ganon-build/data/SEQ2.fna.gz create mode 100644 tests/ganon-build/data/SEQ20.fna.gz create mode 100644 tests/ganon-build/data/SEQ21.fna.gz create mode 100644 tests/ganon-build/data/SEQ22.fna.gz create mode 100644 tests/ganon-build/data/SEQ23.fna.gz create mode 100644 tests/ganon-build/data/SEQ24.fna.gz create mode 100644 tests/ganon-build/data/SEQ3.fna.gz create mode 100644 tests/ganon-build/data/SEQ4.fna.gz create mode 100644 tests/ganon-build/data/SEQ5.fna.gz create mode 100644 tests/ganon-build/data/SEQ6.fna.gz create mode 100644 tests/ganon-build/data/SEQ7.fna.gz create mode 100644 tests/ganon-build/data/SEQ8.fna.gz create mode 100644 tests/ganon-build/data/SEQ9.fna.gz delete mode 100644 tests/ganon-build/data/mode.fasta.gz diff --git a/.travis.yml b/.travis.yml index 936049d3..fafde3d4 100755 --- a/.travis.yml +++ b/.travis.yml @@ -43,7 +43,7 @@ matrix: before_install: - eval "${MATRIX_EVAL}" - python3 -m pip install --upgrade pip - - python3 -m pip install "pandas>=1.1.0" + - python3 -m pip install "pandas>=1.2.0" - python3 -m pip install "multitax>=1.3.1" - if [ "$BUILD_TYPE" == "Coverage" ]; then python3 -m pip install coverage; diff --git a/docs/classification.md b/docs/classification.md index 6dea00af..28f3b976 100644 --- a/docs/classification.md +++ b/docs/classification.md @@ -12,7 +12,7 @@ Output files: - `results.rep`: plain report of the run, used to further generate tree-like reports - `results.tre`: tree-like report with cumulative abundances by taxonomic ranks (can be re-generated with `ganon report`) -By default, `ganon classify` only write report files. To get files with the classification of each read, use `--output-one` and/or `--output-all`. More information about output files [here](../outputfiles/#ganon-classify). +By default, `ganon classify` only write report files. To get files with the classification of each read, use `--output-one` and/or `--output-all`. More information about output files [here](outputfiles.md#ganon-classify). !!! Note ganon performs **taxonomic profiling** and/or **binning** (one tax. assignment for each read) at a taxonomic, strain or sequence level. Some guidelines are listed below, please choose the parameters according to your application. @@ -23,7 +23,7 @@ By default, `ganon classify` only write report files. To get files with the clas - strict thresholds: `--rel-cutoff 0.75` and `--rel-filter 0.1` - `--min-count 0.00005` (0.005%) to exclude very low abundant taxa - - `--report-type abundance` to generate taxonomic abundances, correcting for genome sizes (more infos [here](../reports/#report-type-report-type)) + - `--report-type abundance` to generate taxonomic abundances, correcting for genome sizes (more infos [here](reports.md#report-type-report-type)) ### Binning @@ -31,7 +31,7 @@ To achieve better results for taxonomic binning or sequence classification, `gan - less strict thresholds: `--rel-cutoff 0.25 --rel-filter 0` - `--min-count 0` reports all taxa with at least one read assigned to it - - `--report-type reads` will report sequence abundances instead of taxonomic abundances (more infos [here](../reports/#report-type-report-type)) + - `--report-type reads` will report sequence abundances instead of taxonomic abundances (more infos [here](reports.md#report-type-report-type)) !!! Tip Database parameters in `ganon build` can also influence your results. Lower `--max-fp` (e.g. 0.1, 0.001) and higher `--kmer-size` (e.g. `23`, `27`) will improve sensitivity of your results at cost of a larger database and memory usage. diff --git a/docs/custom_databases.md b/docs/custom_databases.md index f8d06b79..27e8def3 100644 --- a/docs/custom_databases.md +++ b/docs/custom_databases.md @@ -1,81 +1,120 @@ # Custom databases -## Default NCBI assembly or sequence accession +Besides the automated download and build (`ganon build`) ganon provides a highly customizable build procedure (`ganon build-custom`) to create databases from local sequence files. The usage of this procedure depends on the configuration of your files: -Besides the automated download and build (`ganon build`) ganon provides a highly customizable build procedure (`ganon build-custom`) to create databases from local sequence files. +- Filename like `GCA_002211645.1_ASM221164v1_genomic.fna.gz`: genomic fasta files in the NCBI standard, with assembly accession in the beginning of the filename. Provide the files with the `--input` parameter. ganon will try to retrieve all necessary information to build the database. +- Headers like `>NC_006297.1 Bacteroides fragilis YCH46 ...`: sequence headers are in the NCBI standard, with sequence accession in after `>` and with a space afterwards (or line break). Provide the files with the `--input` parameter and set `--input-target sequence`. ganon will try to retrieve all necessary information to build the database. +- For non-standard filenames and headers, [follow this](#non-standard-filesheaders-with-input-file) -To use custom sequences, just provide them with `--input`. ganon will try to retrieve all necessary information necessary to build a database. +!!! warning + `--input-target sequence` will be slower to build and will use more disk space, since files have be re-written separately for each sequence. More information about building by file or sequence can be found [here](#target-file-or-sequence-input-target). -!!! note - ganon expects assembly accessions in the filename like `GCA_002211645.1_ASM221164v1_genomic.fna.gz`. When using `--input-target sequence` filenames are not important but sequence headers should contain sequence accessions like `>CP022124.1 Fusobacterium nu...`. More information about building by file or sequence can be found [here](#target-file-or-sequence-input-target). +The `--level` is a important parameter that will define the (max.) classification level for the database ([more infos](#build-level-level)): + +- `--level file` or `sequence` -> default behavior (depending on `--input-target `), use file/sequence as classification target +- `--level assembly` -> will retrieve assembly related to the file/sequence, use assembly as classification target +- `--level leaves` or `species`,`genus`,... -> group input by taxonomy, use tax. nodes at the rank chosen as classification target -## Non-standard/custom accessions +More infos about other parameters [here](#parameter-details). -It is also possible to use **non-standard accessions and headers** to build custom databases with `--input-file`. This file should contain the following fields (tab-separated): `file [ target node specialization specialization_name].` Note that file is mandatory and additional fields not. +## Non-standard files/headers with `--input-file` -!!! tip - If you just want to build a database without any taxonomic or target information, just sent the files with `--input`, use `--taxonomy skip` and choose between `--input-target file` or `sequence`. +Alternatively to the automatic input methods, it is possible to manually define the input with either standard or **non-standard filenames, accessions and headers** to build custom databases with `--input-file`. This file should contain the following fields (tab-separated): + +`file [ target node specialization specialization_name].` + +- `file`: relative or full path to the sequence file +- `target`: any unique text to name the file, to be used in the taxonomy +- `node`: taxonomic node (e.g. taxid) to link entry with taxonomy +- `specialization`: creates a specialized taxonomic level with a custom name, allowing files to be grouped +- `specialization_name`: a name for the specialization, to be used in the taxonomy !!! warning - the target and specialization fields (2nd and 4th col) cannot be the same as the node (3rd col) + the `target` and `specialization` fields (2nd and 4th col) cannot be the same as the `node` (3rd col) + +Below you find example of `--input-file`. Note they are slightly different depending on the `--input-target` chosen. They need to be *tab-separated* to be properly parsed (tsv). -
- Examples of --input-file -
+### Examples of `--input-file` using the default `--input-target file` -With --input-target file (default), where my_target_1 and my_target_2 are just names to assign sequences from (unique) sequence files: +#### List of files ``` -sequences.fasta my_target_1 -others.fasta my_target_2 +sequences.fasta +others.fasta ``` -With --input-target sequence, second column should match sequence headers on provided sequence files (that should be repeated for each header): +No taxonomic information is provided so `--taxonomy skip` should be set. The classification against the generated database will be performed at file level (`--level file`), since that is the only available information given. +#### List of files with alternative names ``` -sequences.fasta HEADER1 -sequences.fasta HEADER2 -sequences.fasta HEADER3 -others.fasta HEADER4 -others.fasta HEADER5 +sequences.fasta sequences +others.fasta others ``` -A third column with taxonomic nodes can be provided to link the data with taxonomy. For example with --taxonomy ncbi: +Just like above, but with a specific name to be used for each file. + +#### Files and taxonomy ``` -sequences.fasta FILE_A 562 -others.fasta FILE_B 623 +sequences.fasta sequences 562 +others.fasta others 623 ``` +The classification max. level against this database will depend on the value set for `--level`: + +- `--level file` -> use the file (named with target) with node as parent +- `--level leaves` or `species`,`genus`,... -> files are grouped by taxonomy + +#### Files, taxonomy and specialization + ``` -sequences.fasta HEADER1 562 -sequences.fasta HEADER2 562 -sequences.fasta HEADER3 562 -others.fasta HEADER4 623 -others.fasta HEADER5 623 +sequences.fasta FILE_A 562 ID44444 Escherichia coli TW10119 +others.fasta FILE_B 623 ID55555 Shigella flexneri 1a ``` -Further specializations can be used to create a additional classification level after the taxonomic leaves. For example (using --level custom): +The classification max. level against this database will depend on the value set for `--level`: + +- `--level custom` -> use the specialization (named with specialization_name) with node as parent +- `--level file` -> use the file (named with target) as a tax. node as parent +- `--level leaves` or `species`,`genus`,... -> files are grouped by taxonomy + +### Examples of `--input-file` using `--input-target sequence` + +To provide a tabular information for every sequence in your files, you need to use the `target` field (2nd col.) of the `--input-file` to input sequence headers. For example: + +#### Sequences and taxonomy ``` -sequences.fasta FILE_A 562 ID44444 Escherichia coli TW10119 -others.fasta FILE_B 623 ID55555 Shigella flexneri 1a +sequences.fasta NZ_CP054001.1 562 +sequences.fasta NZ_CP117955.1 623 +others.fasta header1 666 +others.fasta header2 666 ``` +The classification max. level against this database will depend on the value set for `--level`: + +- `--level sequence` -> use the sequence header with node as parent +- `--level assembly` -> will attempt to retrieve the assembly related to the sequence with node as parent +- `--level leaves` or `species`,`genus`,... -> files are grouped by taxonomy + +#### Sequences, taxonomy and specialization + ``` -sequences.fasta HEADER1 562 ID443 Escherichia coli TW10119 -sequences.fasta HEADER2 562 ID297 Escherichia coli PCN079 -sequences.fasta HEADER3 562 ID8873 Escherichia coli P0301867.7 -others.fasta HEADER4 623 ID2241 Shigella flexneri 1a -others.fasta HEADER5 623 ID4422 Shigella flexneri 1b +sequences.fasta NZ_CP054001.1 562 ID44444 Escherichia coli TW10119 +sequences.fasta NZ_CP117955.1 623 ID55555 Shigella flexneri 1a +others.fasta header1 666 StrainA My Strain +others.fasta header2 666 StrainA My Strain ``` -
-
+The classification max. level against this database will depend on the value set for `--level`: + +- `--level custom` -> use the specialization (named with specialization_name) with node as parent +- `--level sequence` -> use the sequence header with node as parent +- `--level leaves` or `species`,`genus`,... -> files are grouped by taxonomy ## Examples -Some examples with download and build commands for custom ganon databases from useful and commonly used repositories and datasets for metagenomics analysis: +Below you will find some examples from commonly used repositories for metagenomics analysis with `ganon build-custom`: ### HumGut @@ -121,15 +160,7 @@ wget -A genomic.fna.gz -m -nd --quiet --show-progress "ftp://ftp.ncbi.nlm.nih.go wget -A genomic.fna.gz -m -nd --quiet --show-progress "ftp://ftp.ncbi.nlm.nih.gov/genomes/refseq/plastid/" wget -A genomic.fna.gz -m -nd --quiet --show-progress "ftp://ftp.ncbi.nlm.nih.gov/genomes/refseq/mitochondrion/" -# Split sequences in files and retrieve taxonomy -mkdir sequences/ -zcat plasmid.* plastid.* mitochondrion.* | awk '$0 ~ ">" {accver=(substr($1,2)); print accver}{print $0 > "sequences/"accver".fna"}' | ganon-get-seq-info.sh -e -i - | awk '{print "sequences/"$1".fna\t"$1"\t"$3}' > ppm.tsv - -# Build ganon database -ganon build-custom --input-file ppm.tsv --db-prefix ppm --level species --threads 16 - -# OPTIONAL Remove temporary folder and downloaded files -rm -rf sequences/ ppm.tsv plasmid.* plastid.* mitochondrion.* +ganon build-custom --input plasmid.* plastid.* mitochondrion.* --db-prefix ppm --level species --threads 8 --input-target sequence ``` ### UniVec, UniVec_core @@ -139,13 +170,13 @@ rm -rf sequences/ ppm.tsv plasmid.* plastid.* mitochondrion.* ```bash # UniVec wget -O "UniVec.fasta" --quiet --show-progress "ftp://ftp.ncbi.nlm.nih.gov/pub/UniVec/UniVec" -echo -e "UniVec.fasta\tUniVec\t81077" > UniVec_Core_ganon_input_file.tsv -ganon build-custom --input-file UniVec_ganon_input_file.tsv --db-prefix UniVec --level leaves --threads 8 +echo -e "UniVec.fasta\tUniVec\t81077" > UniVec_ganon_input_file.tsv +ganon build-custom --input-file UniVec_ganon_input_file.tsv --db-prefix UniVec --level leaves --threads 8 --skip-genome-size # UniVec_Core wget -O "UniVec_Core.fasta" --quiet --show-progress "ftp://ftp.ncbi.nlm.nih.gov/pub/UniVec/UniVec_Core" echo -e "UniVec_Core.fasta\tUniVec_Core\t81077" > UniVec_Core_ganon_input_file.tsv -ganon build-custom --input-file UniVec_Core_ganon_input_file.tsv --db-prefix UniVec_Core --level leaves --threads 8 +ganon build-custom --input-file UniVec_Core_ganon_input_file.tsv --db-prefix UniVec_Core --level leaves --threads 8 --skip-genome-size ``` !!! note @@ -155,12 +186,15 @@ ganon build-custom --input-file UniVec_Core_ganon_input_file.tsv --db-prefix Uni "Genome catalogues are biome-specific collections of metagenomic-assembled and isolate genomes". [Article](https://ftp.ebi.ac.uk/pub/databases/metagenomics/mgnify_genomes/)/[Website](https://www.ebi.ac.uk/metagenomics/browse/genomes)/[FTP](https://ftp.ebi.ac.uk/pub/databases/metagenomics/mgnify_genomes/). -There are currently (2023-05-04) 8 genome catalogues available: chicken-gut, human-gut, human-oral, marine, non-model-fish-gut, pig-gut and zebrafish-fecal. An example below how to download and build the human-oral catalog: +Currently available genome catalogues (2024-02-09): `chicken-gut` `cow-rumen` `honeybee-gut` `human-gut` `human-oral` `human-vaginal` `marine` `mouse-gut` `non-model-fish-gut` `pig-gut` `zebrafish-fecal` +*List currently available entries `curl --silent --list-only ftp://ftp.ebi.ac.uk/pub/databases/metagenomics/mgnify_genomes/`* + +Example on how to download and build the `human-oral` catalog: ```bash # Download metadata -wget "https://ftp.ebi.ac.uk/pub/databases/metagenomics/mgnify_genomes/human-oral/v1.0/genomes-all_metadata.tsv" +wget "https://ftp.ebi.ac.uk/pub/databases/metagenomics/mgnify_genomes/human-oral/v1.0.1/genomes-all_metadata.tsv" # Download sequence files with 12 threads tail -n+2 genomes-all_metadata.tsv | cut -f 1,20 | xargs -P 12 -n2 sh -c 'curl --silent ${1}| gzip -d | sed -e "1,/##FASTA/ d" | gzip > ${0}.fna.gz' @@ -169,7 +203,7 @@ tail -n+2 genomes-all_metadata.tsv | cut -f 1,20 | xargs -P 12 -n2 sh -c 'curl - tail -n+2 genomes-all_metadata.tsv | cut -f 1,15 | tr ';' '\t' | awk -F"\t" '{tax="1";for(i=NF;i>1;i--){if(length($i)>3){tax=$i;break;}};print $1".fna.gz\t"$1"\t"tax}' > ganon_input_file.tsv # Build ganon database -ganon build-custom --input-file ganon_input_file.tsv --db-prefix mgnify_human_oral_v1 --taxonomy gtdb --level leaves --threads 32 +ganon build-custom --input-file ganon_input_file.tsv --db-prefix mgnify_human_oral_v101 --taxonomy gtdb --level leaves --threads 8 ``` !!! note @@ -196,15 +230,14 @@ ganon build-custom --input download/ --input-recursive --db-prefix fdaargos --nc BLAST databases. [Website](https://blast.ncbi.nlm.nih.gov/Blast.cgi)/[FTP](https://ftp.ncbi.nlm.nih.gov/blast/db/). -Current available nucleotide databases (2023-05-04): `16S_ribosomal_RNA` `18S_fungal_sequences` `28S_fungal_sequences` `Betacoronavirus` `env_nt` `human_genome` `ITS_eukaryote_sequences` `ITS_RefSeq_Fungi` `LSU_eukaryote_rRNA` `LSU_prokaryote_rRNA` `mito` `mouse_genome` `nt` `nt_euk` `nt_others` `nt_prok` `nt_viruses` `patnt` `pdbnt` `ref_euk_rep_genomes` `ref_prok_rep_genomes` `refseq_rna` `refseq_select_rna` `ref_viroids_rep_genomes` `ref_viruses_rep_genomes` `SSU_eukaryote_rRNA` `tsa_nt` +Current available nucleotide databases (2024-02-09): `16S_ribosomal_RNA` `18S_fungal_sequences` `28S_fungal_sequences` `Betacoronavirus` `env_nt` `human_genome` `ITS_eukaryote_sequences` `ITS_RefSeq_Fungi` `LSU_eukaryote_rRNA` `LSU_prokaryote_rRNA` `mito` `mouse_genome` `nt` `nt_euk` `nt_others` `nt_prok` `nt_viruses` `patnt` `pdbnt` `ref_euk_rep_genomes` `ref_prok_rep_genomes` `refseq_rna` `refseq_select_rna` `ref_viroids_rep_genomes` `ref_viruses_rep_genomes` `SSU_eukaryote_rRNA` `tsa_nt` -!!! note - List currently available nucleotide databases `curl --silent --list-only ftp://ftp.ncbi.nlm.nih.gov/blast/db/ | grep "nucl-metadata.json" | sed 's/-nucl-metadata.json/, /g' | sort` +*List currently available entries `curl --silent --list-only ftp://ftp.ncbi.nlm.nih.gov/blast/db/ | grep "nucl-metadata.json" | sed 's/-nucl-metadata.json/, /g' | sort`* !!! warning - Some BLAST databases are very big and may require extreme computational resources to build. You may need to use some [reduction strategies](../default_databases/#reducing-database-size) + Some BLAST databases are very big and may require extreme computational resources to build. You may need to use some [reduction strategies](default_databases.md#reducing-database-size). -The example below extracts sequences and information from a BLAST db to build a ganon database: +The example shows how to **download**, **parse** and **build** a ganon database from BLAST database files. It does so by splitting the database into taxonomic specific files, to speed-up the build process: ```bash # Define BLAST db @@ -226,18 +259,18 @@ find -name "${db}.*tar.gz" -type f -printf '%P\n' | xargs -P ${threads} -I{} sh seq 0 9 | xargs -i mkdir -p "${db}"/{} # This command extracts sequences from the blastdb and writes them into taxid specific files -# It also generates the --input-file for ganon +# It also generates the --input-file for ganon with the fields: filepath file taxid blastdbcmd -entry all -db "${db}" -outfmt "%a %T %s" | \ -awk -v db="$(realpath ${db})" '{file=db"/"substr($2,1,1)"/"$2".fna"; print ">"$1"\n"$3 >> file; print file"\t"$2"\t"$2}' | \ +awk -v db="$(realpath ${db})" '{file=db"/"substr($2,1,1)"/"$2".fna"; print ">"$1"\n"$3 >> file; print file"\t"$2".fna\t"$2}' | \ sort | uniq > "${db}_ganon_input_file.tsv" # Build ganon database -ganon build-custom --input-file "${db}_ganon_input_file.tsv" --db-prefix "${db}" --threads 12 +ganon build-custom --input-file "${db}_ganon_input_file.tsv" --db-prefix "${db}" --threads ${threads} --level leaves # Delete extracted files and auxiliary files cat "${db}_extracted_files.txt" | xargs rm rm "${db}_extracted_files.txt" "${db}.md5" "${db}_downloaded.md5" -# Delete sequences +# Delete sequences and input_file rm -rf "${db}" "${db}_ganon_input_file.tsv" ``` @@ -246,7 +279,7 @@ rm -rf "${db}" "${db}_ganon_input_file.tsv" ### Files from genome_updater -To create a ganon database from files previosly downloaded with [genome_updater](https://github.com/pirovc/genome_updater): +To create a ganon database from files previously downloaded with [genome_updater](https://github.com/pirovc/genome_updater): ```bash ganon build-custom --input output_folder_genome_updater/version/ --input-recursive --db-prefix mydb --ncbi-file-info output_folder_genome_updater/assembly_summary.txt --level assembly --threads 32 @@ -256,7 +289,7 @@ ganon build-custom --input output_folder_genome_updater/version/ --input-recursi ### False positive and size (--max-fp, --filter-size) -ganon indices are based on bloom filters and can have false positive matches. This can be controlled with `--max-fp` parameter. The lower the `--max-fp`, the less chances of false positives matches on classification, but the larger the database size will be. For example, with `--max-fp 0.01` the database will be build so any target (defined by `--level`) will have 1 in a 100 change of reporting a false k-mer match. [The false positive of the query](../classification/#false-positive-of-a-query-fpr-query) (all k-mers of a read) will be way lower, but directly affected by this value. +ganon indices are based on bloom filters and can have false positive matches. This can be controlled with `--max-fp` parameter. The lower the `--max-fp`, the less chances of false positives matches on classification, but the larger the database size will be. For example, with `--max-fp 0.01` the database will be build so any target (defined by `--level`) will have 1 in a 100 change of reporting a false k-mer match. [The false positive of the query](classification.md#false-positive-of-a-query-fpr-query) (all k-mers of a read) will be way lower, but directly affected by this value. Alternatively, one can set a specific size for the final index with `--filter-size`. When using this option, please observe the theoretic false positive of the index reported at the end of the building process. @@ -266,15 +299,19 @@ in `ganon build`, when `--window-size` > `--kmer-size` minimizers are used. That ### Target file or sequence (--input-target) -Customized builds can be done either by file or sequence. `--input-target file` will consider every file provided with `--input` a single unit. `--input-target sequence` will use every sequence as a unit. +This is a parameter that defines how ganon will parse your input files: + - `--input-target file` (default) will consider every file provided with `--input` a single unit (e.g. multi-fasta files are considered one input, sequence headers ignored). + - `--input-target sequence` will use every sequence as a unit. For this, ganon will first decompose every sequence in the input files provided with `--input` into a separated file. This will take longer and use more disk space. -`--input-target file` is the default behavior and most efficient way to build databases. `--input-target sequence` should only be used when the input sequences are stored in a single file or when classification at sequence level is desired. +`--input-target file` is the default behavior and most efficient way to build databases. `--input-target sequence` should only be used when the input sequences are not separated by file (e.g. a single big FASTA file) or when classification at sequence level is desired. ### Build level (--level) -The `--level` parameter defines the max. depth of the database for classification. This parameter is relevant because the `--max-fp` is going to be guaranteed at the `--level` chosen. By default, the level will be the same as `--input-target`, meaning that classification will be done either at file or sequence level. +The `--level` parameter defines the max. depth of the database for classification. This parameter is relevant because the `--max-fp` is going to be guaranteed at the `--level` chosen. + +In `ganon build` the default value is `species`. In `ganon build-custom` the level will be the same as `--input-target`, meaning that classification will be done either at `file` or `sequence` level. -Alternatively, `--level assembly` will link the file or sequence target information with assembly accessions retrieved from NCBI servers. `--level leaves` or `--level species` (or genus, family, ...) will link the targets with taxonomic information and prune the tree at the chosen level. `--level custom` will use specialization level define in the `--input-file`. +Alternatively, `--level assembly` will link the file or sequence target information with assembly accessions retrieved from NCBI. `--level leaves` or `--level species` (or `genus`, `family`, ...) will link the targets with taxonomic information and prune the tree at the chosen level. `--level custom` will use specialization (4th col.) defined in the `--input-file`. ### Genome sizes (--genome-size-files) diff --git a/docs/default_databases.md b/docs/default_databases.md index 7577c178..cec3b8ba 100644 --- a/docs/default_databases.md +++ b/docs/default_databases.md @@ -13,7 +13,7 @@ This will download archaeal and bacterial complete genomes from RefSeq and build ganon update -d arc_bac -t 30 ``` -Additionally, [custom databases](../custom_databases) can be built with customized files and identifiers with the `ganon build-custom` command. +Additionally, [custom databases](custom_databases.md) can be built with customized files and identifiers with the `ganon build-custom` command. !!! info We DO NOT provide pre-built indices for download. ganon can build databases very efficiently. This way, you will always have up-to-date reference sequences and get most out of your data. @@ -56,9 +56,9 @@ NCBI RefSeq and GenBank repositories are common resources to obtain reference se - As a rule of thumb, the more the better, so choose the most comprehensive sub-set as possible given your computational resources - It is possible to build databases that consume a fixed size/RAM usage. Beware that smaller filters will increase the false positive rates when classifying. Other approaches [can reduce the size/RAM requirements with some trade-offs](#reducing-database-size). -- Alternatively, you can build one database for each organism group separately and use them in `ganon classify` in [any order or even stack them hierarchically](../classification/#multiple-and-hierarchical-classification). This way combination of multiple databases are possible, extending use cases. +- Alternatively, you can build one database for each organism group separately and use them in `ganon classify` in [any order or even stack them hierarchically](classification.md#multiple-and-hierarchical-classification). This way combination of multiple databases are possible, extending use cases. -Further examples of commonly used database can be found [here](../custom_databases/#examples). +Further examples of commonly used database can be found [here](custom_databases.md#examples). ### Specific organisms or taxonomic groups @@ -82,7 +82,7 @@ will download top 3 archaeal assemblies for each genus with date before 2023-01- ## GTDB -By default, ganon will use the NCBI Taxonomy to build the database. However, [GTDB](gtdb.ecogenomic.org/) is fully supported and can be used with the parameter `--taxonomy gtdb`. +By default, ganon will use the NCBI Taxonomy to build the database. However, [GTDB](https://gtdb.ecogenomic.org) is fully supported and can be used with the parameter `--taxonomy gtdb`. Filtering by taxonomic entries also work with GTDB, for example: @@ -121,7 +121,7 @@ genome_updater.sh -e assembly_summary.txt -f "genomic.fna.gz" -o recovered_files ### Filter type (IBF and HIBF) -The Hierarchical Interleaved Bloom Filter (HIBF) is an improvement over the default Interleaved Bloom Filter (IBF) and generates *smaller* databases with *faster* query times ([article](https://genomebiology.biomedcentral.com/articles/10.1186/s13059-023-02971-4)). However, the HIBF takes longer to build and has less flexibility regarding size and further options in ganon. You can choose which filter to use with the `--filter-type` parameter in `ganon build` and `ganon build-custom`- +The Hierarchical Interleaved Bloom Filter (HIBF) is an improvement over the default Interleaved Bloom Filter (IBF) and generates *smaller* databases with *faster* query times ([article](https://genomebiology.biomedcentral.com/articles/10.1186/s13059-023-02971-4)). However, the HIBF takes a little longer to build and has less flexibility regarding size and further options in ganon. You can choose which filter to use with the `--filter-type` parameter in `ganon build` and `ganon build-custom`. Due to differences between the default IBF used in ganon and the HIBF, it is recommended to lower the false positive when using the HIBF. The default value for high sensitivity is 1% (`--filter-type hibf --max-fp 0.001`). @@ -129,10 +129,9 @@ Due to differences between the default IBF used in ganon and the HIBF, it is rec - For large unbalanced reference sets, lots of reads to query -> HIBF (default) - For quick database build and more flexibility -> IBF - ### False positive rate -A higher `--max-fp` value will generate a smaller database but with a higher number of false positive matches on classification. [More details](../custom_databases/#false-positive-and-size-max-fp-filter-size). Values between `0.001` (0.1%) and `0.3` (30%) are generally used. +A higher `--max-fp` value will generate a smaller database but with a higher number of false positive matches on classification. [More details](custom_databases.md#false-positive-and-size-max-fp-filter-size). Values between `0.001` (0.1%) and `0.3` (30%) are generally used. !!! hint When using higher `--max-fp` values, more false positive results may be generated. This can be filtered with the `--fpr-query` parameter in `ganon classify` @@ -140,7 +139,7 @@ A higher `--max-fp` value will generate a smaller database but with a higher num ### k-mer and window size -Define how much unique information is stored in the database. [More details](../custom_databases/#minimizers-window-size-kmer-size) +Define how much unique information is stored in the database. [More details](custom_databases.md#minimizers-window-size-kmer-size) - The smaller the `--kmer-size`, the less unique they will be, reducing database size but also sensitivity in classification. - The bigger the `--window-size`, the less information needs to be stored resulting in smaller databases but with decrease classification accuracy. @@ -158,7 +157,7 @@ RefSeq and GenBank are highly biased toward some few organisms. This means that ### Split databases -Ganon allows classification with multiple databases in one level or in an hierarchy ([More details](../classification/#multiple-and-hierarchical-classification)). This means that databases can be built separately and used in any combination as desired. There are usually some benefits of doing so: +Ganon allows classification with multiple databases in one level or in an hierarchy ([More details](classification.md#multiple-and-hierarchical-classification)). This means that databases can be built separately and used in any combination as desired. There are usually some benefits of doing so: - Smaller databases when building by organism group, for example: one for bacteria, another for viruses, ... since average genome sizes are quite different. - Easier to maintain and update. @@ -168,7 +167,7 @@ Ganon allows classification with multiple databases in one level or in an hierar ### Fixed size and Mode (only for --filter-type ibf) -A fixed size for the database filter can be defined with `--filter-size` when using `--filter-type ibf`. The smaller the filter size, the higher the false positive chances on classification. When using a fixed filter size, ganon will report the max. and avg. false positive rate at the end of the build. [More details](../custom_databases/#false-positive-and-size-max-fp-filter-size). +A fixed size for the database filter can be defined with `--filter-size` when using `--filter-type ibf`. The smaller the filter size, the higher the false positive chances on classification. When using a fixed filter size, ganon will report the max. and avg. false positive rate at the end of the build. [More details](custom_databases.md#false-positive-and-size-max-fp-filter-size). `--mode` offers 5 different categories to build a database controlling the trade-off between size and classification speed. diff --git a/docs/index.md b/docs/index.md index dbfe57e9..59571515 100644 --- a/docs/index.md +++ b/docs/index.md @@ -10,17 +10,17 @@ ganon is designed to index large sets of genomic reference sequences and to clas ## Features -- integrated download and build of any subset from [RefSeq/Genbank/GTDB](default_databases/#refseq-and-genbank) with incremental [updates](default_databases/#update-ganon-update) -- NCBI and [GTDB](default_databases/#gtdb) native support for taxonomic classification, custom taxonomy or no taxonomy at all -- [customizable database](custom_databases/) build for local or non-standard sequence files -- optimized [taxonomic binning](classification/#binning) and [profiling](classification/#profiling) configurations +- integrated download and build of any subset from [RefSeq/Genbank/GTDB](default_databases.md#refseq-and-genbank) with incremental [updates](default_databases.md#update-ganon-update) +- NCBI and [GTDB](default_databases.md#gtdb) native support for taxonomic classification, custom taxonomy or no taxonomy at all +- [customizable database](custom_databases.md) build for local or non-standard sequence files +- optimized [taxonomic binning](classification.md#binning) and [profiling](classification.md#profiling) configurations - build and classify at various taxonomic levels, strain, assembly, file, sequence or custom specialization -- [hierarchical classification](classification/#multiple-and-hierarchical-classification) using several databases in one or more levels in just one run -- [EM and/or LCA](classification/#reads-with-multiple-matches) algorithms to solve multiple-matching reads +- [hierarchical classification](classification.md#multiple-and-hierarchical-classification) using several databases in one or more levels in just one run +- [EM and/or LCA](classification.md#reads-with-multiple-matches) algorithms to solve multiple-matching reads - reporting of multiple and unique matches for every read -- [reporting](reports/#report-type-report-type) of sequence, taxonomic or multi-match abundances with optional genome size correction -- advanced tree-like [reports](reports) with several filter options -- generation of [contingency tables](table/) with several filters for multi-sample studies +- [reporting](reports.md#report-type-report-type) of sequence, taxonomic or multi-match abundances with optional genome size correction +- advanced tree-like [reports](reports.md) with several filter options +- generation of [contingency tables](table.md) with several filters for multi-sample studies ganon achieved very good results in [our own evaluations](https://dx.doi.org/10.1093/bioinformatics/btaa458) but also in independent evaluations: [LEMMI](https://lemmi-v1.ezlab.org/), [LEMMI v2](https://lemmi.ezlab.org/) and [CAMI2](https://dx.doi.org/10.1038/s41592-022-01431-4) @@ -39,7 +39,7 @@ However, there are possible performance benefits compiling ganon from source in ### Python dependencies - python >=3.6 -- pandas >=1.1.0 +- pandas >=1.2.0 - [multitax](https://github.com/pirovc/multitax) >=1.3.1 ```bash @@ -48,9 +48,9 @@ python3 -V # Install packages via pip or conda: # PIP -python3 -m pip install "pandas>=1.1.0" "multitax>=1.3.1" -# Conda (alternative) -conda install "pandas>=1.1.0" "multitax>=1.3.1" +python3 -m pip install "pandas>=1.2.0" "multitax>=1.3.1" +# Conda/Mamba (alternative) +conda install "pandas>=1.2.0" "multitax>=1.3.1" ``` ### C++ dependencies diff --git a/docs/reports.md b/docs/reports.md index 9c03b8e4..0572512f 100644 --- a/docs/reports.md +++ b/docs/reports.md @@ -40,7 +40,7 @@ Several reports are available with `--report-type`: `reads`, `abundance`, `dist` `reads` reports **sequence abundances** which are the basic proportion of reads classified in the sample. -`abundance` will convert sequence abundance into **taxonomic abundances** by re-distributing read counts among leaf nodes and correcting by genome size. The re-distribution applies for reads classified with a LCA assignment and it is proportional to the number of unique matches of leaf nodes available in the ganon database (relative to the LCA node). Genome size is estimated based on [NCBI or GTDB auxiliary files](../custom_databases/#genome-sizes-genome-size-files). Genome size correction is applied by rank based on default ranks only (superkingdom phylum class order family genus species assembly). Read counts in intermediate ranks will be corrected based on the closest parent default rank and re-assigned to its original rank. +`abundance` will convert sequence abundance into **taxonomic abundances** by re-distributing read counts among leaf nodes and correcting by genome size. The re-distribution applies for reads classified with a LCA assignment and it is proportional to the number of unique matches of leaf nodes available in the ganon database (relative to the LCA node). Genome size is estimated based on [NCBI or GTDB auxiliary files](custom_databases.md#genome-sizes-genome-size-files). Genome size correction is applied by rank based on default ranks only (superkingdom phylum class order family genus species assembly). Read counts in intermediate ranks will be corrected based on the closest parent default rank and re-assigned to its original rank. `dist` is the same of `reads` with read count re-distribution diff --git a/src/ganon-build/CommandLineParser.cpp b/src/ganon-build/CommandLineParser.cpp index bfeadd0e..e933388e 100644 --- a/src/ganon-build/CommandLineParser.cpp +++ b/src/ganon-build/CommandLineParser.cpp @@ -13,7 +13,7 @@ std::optional< Config > CommandLineParser::parse( int argc, char** argv ) // clang-format off options.add_options() - ( "i,input-file", "Define sequences to use. Tabular file with the fields: file [ target seqid]", cxxopts::value< std::string >() ) + ( "i,input-file", "Define sequences to use. Tabular file with the fields: file [ target]", cxxopts::value< std::string >() ) ( "o,output-file", "Filter output file", cxxopts::value< std::string >() ) ( "k,kmer-size", "k-mer size. Default: 19", cxxopts::value< uint8_t >() ) ( "w,window-size", "window size. Default: 31", cxxopts::value< uint16_t >() ) diff --git a/src/ganon-build/GanonBuild.cpp b/src/ganon-build/GanonBuild.cpp index da9121da..4fa58ae1 100644 --- a/src/ganon-build/GanonBuild.cpp +++ b/src/ganon-build/GanonBuild.cpp @@ -38,15 +38,14 @@ namespace detail typedef seqan3::interleaved_bloom_filter< seqan3::data_layout::uncompressed > TIBF; -typedef robin_hood::unordered_map< std::string, robin_hood::unordered_set< std::string > > TFile; -typedef robin_hood::unordered_map< std::string, uint64_t > THashesCount; +typedef robin_hood::unordered_map< std::string, uint64_t > THashesCount; typedef robin_hood::unordered_map< uint64_t, std::tuple< std::string, uint64_t, uint64_t > > TBinMapHash; struct InputFileMap { - std::string target; - TFile files; + std::string target; + std::vector< std::string > files; }; @@ -84,10 +83,10 @@ inline std::string get_seqid( std::string header ) return header.substr( 0, header.find( ' ' ) ); } -robin_hood::unordered_map< std::string, TFile > parse_input_file( const std::string& input_file, - THashesCount& hashes_count, - bool quiet, - Stats& stats ) +robin_hood::unordered_map< std::string, std::vector< std::string > > parse_input_file( const std::string& input_file, + THashesCount& hashes_count, + bool quiet, + Stats& stats ) { /* * Funtion to parse input file -> tabular file with the fields: file [ target seqid] @@ -95,10 +94,10 @@ robin_hood::unordered_map< std::string, TFile > parse_input_file( const std::str * In case of sequence parsing (provided seqids on third row), seqids is a set of sequence ids * In case of file parsing, each file has only one target and seqids is empty */ - robin_hood::unordered_map< std::string, TFile > input_map; - std::string line; - robin_hood::unordered_set< std::string > files; - std::ifstream infile( input_file ); + robin_hood::unordered_map< std::string, std::vector< std::string > > input_map; + std::string line; + robin_hood::unordered_set< std::string > files; + std::ifstream infile( input_file ); while ( std::getline( infile, line, '\n' ) ) { std::istringstream stream_line( line ); @@ -120,21 +119,14 @@ robin_hood::unordered_map< std::string, TFile > parse_input_file( const std::str if ( fields.size() == 1 ) { // target is the file itself (filename only without path) - const auto target = std::filesystem::path( file ).filename(); - input_map[target][file] = {}; - hashes_count[target] = 0; + const auto target = std::filesystem::path( file ).filename(); + input_map[target].push_back( file ); + hashes_count[target] = 0; } else if ( fields.size() == 2 ) { // provided target in the file - input_map[fields[1]][file] = {}; - hashes_count[fields[1]] = 0; - } - else - { - // parse by sequence - const std::string seqid = fields[2]; - input_map[fields[1]][file].insert( seqid ); + input_map[fields[1]].push_back( file ); hashes_count[fields[1]] = 0; } } @@ -217,7 +209,7 @@ void count_hashes( SafeQueue< InputFileMap >& ifm_queue, break; // For all files of a target - for ( auto& [file, seqids] : ifm.files ) + for ( auto& file : ifm.files ) { try { @@ -226,88 +218,26 @@ void count_hashes( SafeQueue< InputFileMap >& ifm_queue, seqan3::fields< seqan3::field::id, seqan3::field::seq > > fin{ file }; - if ( seqids.empty() ) - { - // File as target - generate all hashes from file with possible multiple sequences - // before counting and storing - robin_hood::unordered_set< uint64_t > hashes; - for ( auto const& [header, seq] : fin ) - { - if ( seq.size() < config.min_length ) - { - total.skipped_sequences++; - continue; - } - - total.sequences++; - total.length_bp += seq.size(); - const auto mh = seq | minimiser_view | std::views::common; - hashes.insert( mh.begin(), mh.end() ); - } - hashes_count[ifm.target] += hashes.size(); - detail::store_hashes( ifm.target, hashes, config.tmp_output_folder ); - } - else if ( seqids.size() == 1 ) + // File as target - generate all hashes from file with possible multiple sequences + // before counting and storing + + robin_hood::unordered_set< uint64_t > hashes; + for ( auto const& [header, seq] : fin ) { - // Single seqid for a target (or target is seqid itself) - // Directly count and store - for ( auto const& [header, seq] : fin ) + if ( seq.size() < config.min_length ) { - const auto seqid = get_seqid( header ); - if ( seqids.count( seqid ) ) - { - if ( seq.size() < config.min_length ) - { - total.skipped_sequences++; - } - else - { - total.sequences++; - total.length_bp += seq.size(); - robin_hood::unordered_set< uint64_t > hashes; - const auto mh = seq | minimiser_view | std::views::common; - hashes.insert( mh.begin(), mh.end() ); - hashes_count[ifm.target] += hashes.size(); - detail::store_hashes( ifm.target, hashes, config.tmp_output_folder ); - } - break; - } + total.skipped_sequences++; + continue; } - } - else - { - // Multiple seqids for a target - // generate all hashes from file with possible multiple sequences before counting and storing - // Keep track of total seqids to be parsed - size_t n_seqids = seqids.size(); - robin_hood::unordered_set< uint64_t > hashes; - for ( auto const& [header, seq] : fin ) - { - const auto seqid = get_seqid( header ); - if ( seqids.count( seqid ) ) - { - if ( seq.size() < config.min_length ) - { - total.skipped_sequences++; - continue; - } - else - { - total.sequences++; - total.length_bp += seq.size(); - const auto mh = seq | minimiser_view | std::views::common; - hashes.insert( mh.begin(), mh.end() ); - } - // In case all seqids were already processed - if ( !--n_seqids ) - break; - } - } - hashes_count[ifm.target] += hashes.size(); - detail::store_hashes( ifm.target, hashes, config.tmp_output_folder ); + total.sequences++; + total.length_bp += seq.size(); + const auto mh = seq | minimiser_view | std::views::common; + hashes.insert( mh.begin(), mh.end() ); } + hashes_count[ifm.target] += hashes.size(); + detail::store_hashes( ifm.target, hashes, config.tmp_output_folder ); } catch ( seqan3::parse_error const& e ) { @@ -856,7 +786,7 @@ bool run( Config config ) detail::parse_input_file( config.input_file, hashes_count, config.quiet, stats ) ) { // Add to Target and Files to the SafeQueue - ifm_queue.push( detail::InputFileMap{ target, std::move( files ) } ); + ifm_queue.push( detail::InputFileMap{ target, files } ); } ifm_queue.notify_push_over(); diff --git a/src/ganon/build_update.py b/src/ganon/build_update.py index d9d6a147..c45a116f 100644 --- a/src/ganon/build_update.py +++ b/src/ganon/build_update.py @@ -25,7 +25,6 @@ from multitax import NcbiTx, GtdbTx - def build(cfg): # Set paths @@ -248,25 +247,20 @@ def build_custom(cfg, which_call: str="build_custom"): # Retrieve and check input files or folders if cfg.input: + tx = time.time() + print_log("Parsing --input", cfg.quiet) input_files = validate_input_files(cfg.input, cfg.input_extension, cfg.quiet, input_recursive=cfg.input_recursive) + print_log(" - done in " + str("%.2f" % (time.time() - tx)) + "s.\n", cfg.quiet) if not input_files: print_log("ERROR: No valid input files found", cfg.quiet) return False - # Set --input-target automatically if not given - if not cfg.input_target: - # file is the default if more than one file is provided - if len(input_files) > 1 or cfg.input_file: - cfg.input_target = "file" - else: - cfg.input_target = "sequence" - # Set-up taxonomy if cfg.taxonomy != "skip": tax = load_taxonomy(cfg, build_output_folder) # Set-up input info - info = load_input(cfg, input_files) + info = load_input(cfg, input_files, build_output_folder) # Define user bins for writing taxonomy and target info file user_bins_col = "target" # Default as target if cfg.level in cfg.choices_level: @@ -323,7 +317,7 @@ def build_custom(cfg, which_call: str="build_custom"): write_info_file(info, cfg.db_prefix + ".info.tsv") # Write aux file for ganon-build - write_target_info(info, cfg.input_target, user_bins_col, target_info_file) + write_target_info(info, user_bins_col, target_info_file) save_state(which_call + "_parse", files_output_folder) # Skip if already finished run @@ -332,9 +326,10 @@ def build_custom(cfg, which_call: str="build_custom"): else: if cfg.filter_type == "hibf": - tx = time.time() print_log("Building index (raptor)", cfg.quiet) + tx = time.time() + print_log("Preparing files", cfg.quiet) # rewrite target_info_file with one line for each target # symbolic link is created for the first time with the target name # this is a workaround to name targets with raptor @@ -348,6 +343,8 @@ def build_custom(cfg, which_call: str="build_custom"): n_bins = len(target_files) raptor_input_file = build_output_folder + "hibf.txt" + + # Create symbolic link with target to raptor with open(raptor_input_file, "w") as filehibf: for target, files in target_files.items(): # raptor v3.0.0 "eats" the . (e.g. GCF_013391805.1 -> GCF_013391805) @@ -368,18 +365,21 @@ def build_custom(cfg, which_call: str="build_custom"): Path(target_file).symlink_to(first_file) # Write input file for raptor (space separated) filehibf.write(target_file + " " + " ".join(files[1:]) + "\n") + print_log(" - done in " + str("%.2f" % (time.time() - tx)) + "s.\n", cfg.quiet) + tx = time.time() print_log("raptor prepare", cfg.quiet) run_raptor_prepare_cmd = " ".join([cfg.path_exec['raptor'], "prepare", - "--input '" + raptor_input_file + "'", - "--output '" + build_output_folder + "'", - "--kmer " + str(cfg.kmer_size), - "--window " + str(cfg.window_size), - "--quiet" if not cfg.verbose else "", - "--threads " + str(cfg.threads)]) + "--input '" + raptor_input_file + "'", + "--output '" + build_output_folder + "'", + "--kmer " + str(cfg.kmer_size), + "--window " + str(cfg.window_size), + "--quiet" if not cfg.verbose else "", + "--threads " + str(cfg.threads)]) run(run_raptor_prepare_cmd, quiet=cfg.quiet) print_log(" - done in " + str("%.2f" % (time.time() - tx)) + "s.\n", cfg.quiet) + tx = time.time() print_log("raptor layout", cfg.quiet) # Use info file as input for raptor run_raptor_layout_cmd = " ".join([cfg.path_exec['raptor'], "layout", @@ -456,7 +456,7 @@ def build_custom(cfg, which_call: str="build_custom"): ######################################################################################################################## -def parse_input_file(input_file, info, input_target): +def parse_input_file(input_file, info_cols, input_target, quiet): """ parse user provided --input-file with all specifications for input """ @@ -464,35 +464,60 @@ def parse_input_file(input_file, info, input_target): sep="\t", header=None, skiprows=0, - dtype=object, - names=["file", "target", "node", "specialization", "specialization_name"]) - + dtype=object) + # Need to rename columns after parsing since file can have 1 to 5 + info.rename(columns=lambda x: info_cols[x], inplace=True) + info = pd.concat([info, pd.DataFrame(columns=info_cols)]) + # If no target was provided and target is file, use filename if info["target"].isna().all() and input_target == "file": info["target"] = info["file"].apply(os.path.basename) + # Validate files provided and remove not found + total_files = len(info["file"].unique().tolist()) + valid_files = validate_input_files(info["file"].unique().tolist(), "", quiet) + if total_files - len(valid_files) > 0: + info = info[info["file"].isin(valid_files)] + print_log(" - " + str(total_files - len(valid_files)) + " invalid files skipped", quiet) return info -def load_input(cfg, input_files): +def load_input(cfg, input_files, build_output_folder): """ Load basic target info, either provided as --target-info or extracted from file/sequences """ tx = time.time() - info = pd.DataFrame(columns=["target", "node", "specialization", "specialization_name", "file"]) + info_cols = ["file", "target", "node", "specialization", "specialization_name"] # Parse/load info without setting index if cfg.input_file: print_log("Parsing --input-file " + cfg.input_file, cfg.quiet) - info = parse_input_file(cfg.input_file, info, cfg.input_target) + info = parse_input_file(cfg.input_file, info_cols, cfg.input_target, cfg.quiet) + + # If --input-target sequence, split files by sequence and validate headers + if cfg.input_target == "sequence": + print_log("\nMatching --input-file entries with sequences", cfg.quiet) + shape_tmp = info.shape[0] + # Split files and return headers obtained + info_seqs = parse_sequence_accession(info["file"].unique().tolist(), info_cols, build_output_folder) + + # Merge with provided file, keeping only matching entries + info = pd.merge(left=info, right=info_seqs, on="target", suffixes=("", "_seqs"))[info_cols + ["file_seqs"]] + info["file"] = info["file_seqs"] + info.drop("file_seqs", axis=1, inplace=True) + + if shape_tmp - info.shape[0] > 0: + print_log(" - " + str(shape_tmp - info.shape[0]) + " invalid sequences skipped", cfg.quiet) + else: + if cfg.input_target == "sequence": print_log("Parsing sequences from --input (" + str(len(input_files)) + " files)", cfg.quiet) - info = parse_sequence_accession(input_files, info) + info = parse_sequence_accession(input_files, info_cols, build_output_folder) else: - print_log("Parsing --input (" + str(len(input_files)) + " files)", cfg.quiet) - info = parse_file_accession(input_files, info) + print_log("Parsing files from --input (" + str(len(input_files)) + " files)", cfg.quiet) + info = parse_file_accession(input_files, info_cols) # Drop cols without values shape_tmp = info.shape[0] @@ -577,17 +602,14 @@ def write_tax(tax_file, info, tax, genome_sizes, user_bins_col, level, input_tar tax_df["genome_size"] = tax_df.apply(lambda d: genome_sizes[d.node] if d.node in genome_sizes else genome_sizes[d.parent], axis=1) tax_df.to_csv(tax_file, sep="\t", header=False, index=False) -def write_target_info(info, input_target, user_bins_col, target_info_file): +def write_target_info(info, user_bins_col, target_info_file): """ - write tabular file to be parsed by ganon-build with: file target [ sequence] + write tabular file to be parsed by ganon-build with: file target """ with open(target_info_file, "w") as outf: for target, row in info.iterrows(): t = row[user_bins_col] if user_bins_col != "target" else target - if input_target == "sequence": - print(row["file"], t, target, sep="\t", end="\n", file=outf) - else: - print(row["file"], t, sep="\t", end="\n", file=outf) + print(row["file"], t, sep="\t", end="\n", file=outf) def write_info_file(info, filename): diff --git a/src/ganon/config.py b/src/ganon/config.py index fd7514d5..0c3f0198 100644 --- a/src/ganon/config.py +++ b/src/ganon/config.py @@ -88,11 +88,11 @@ def __init__(self, which: str=None, **kwargs): build_custom_required_args.add_argument("-c", "--input-recursive", action="store_true", help="Look for files recursively in folder(s) provided with --input") build_custom_args = build_custom_parser.add_argument_group("custom arguments") - build_custom_args.add_argument("-n", "--input-file", type=file_exists, metavar="", help="Manually set information for input files: file [target node specialization specialization name]. target is the sequence identifier if --input-target sequence (file can be repeated for multiple sequences). if --input-target file and target is not set, filename is used. node is the taxonomic identifier. Mutually exclusive --input") - build_custom_args.add_argument("-a", "--input-target", type=str, metavar="", help="Target to use [file, sequence]. By default: 'file' if multiple input files are provided or --input-file is set, 'sequence' if a single file is provided. Using 'file' is recommended and will speed-up the building process", choices=self.choices_input_target) - build_custom_args.add_argument("-l", "--level", type=str, metavar="", help="Use a specialized target to build the database. By default, --level is the --input-target. Options: any available taxonomic rank [species, genus, ...] or 'leaves' (requires --taxonomy). Further specialization options [" + ", ".join(self.choices_level) + "]. assembly will retrieve and use the assembly accession and name. custom requires and uses the specialization field in the --input-file.") - build_custom_args.add_argument("-m", "--taxonomy-files", type=file_exists, nargs="*", metavar="", help="Specific files for taxonomy - otherwise files will be downloaded") - build_custom_args.add_argument("-z", "--genome-size-files", type=file_exists, nargs="*", metavar="", help="Specific files for genome size estimation - otherwise files will be downloaded") + build_custom_args.add_argument("-n", "--input-file", type=file_exists, metavar="", help="Manually set information for input files: file [target node specialization specialization name]. target is the sequence identifier if --input-target sequence (file can be repeated for multiple sequences). if --input-target file and target is not set, filename is used. node is the taxonomic identifier. Mutually exclusive --input") + build_custom_args.add_argument("-a", "--input-target", type=str, default="file", metavar="", help="Target to use [file, sequence]. Parse input by file or by sequence. Using 'file' is recommended and will speed-up the building process", choices=self.choices_input_target) + build_custom_args.add_argument("-l", "--level", type=str, metavar="", help="Max. level to build the database. By default, --level is the --input-target. Options: any available taxonomic rank [species, genus, ...] or 'leaves' (requires --taxonomy). Further specialization options [" + ", ".join(self.choices_level) + "]. assembly will retrieve and use the assembly accession and name. custom requires and uses the specialization field in the --input-file.") + build_custom_args.add_argument("-m", "--taxonomy-files", type=file_exists, nargs="*", metavar="", help="Specific files for taxonomy - otherwise files will be downloaded") + build_custom_args.add_argument("-z", "--genome-size-files", type=file_exists, nargs="*", metavar="", help="Specific files for genome size estimation - otherwise files will be downloaded") build_custom_args.add_argument("--skip-genome-size", action="store_true", help="Do not attempt to get genome sizes. Activate this option when using sequences not representing full genomes.") ncbi_args = build_custom_parser.add_argument_group("ncbi arguments") @@ -372,10 +372,6 @@ def validate(self): print_log("--input-file is mutually exclusive with --input") return False - if self.filter_type == "hibf" and self.input_target == "sequence": - print_log("--filter-type hibf is currently only supported with --input-target file") - return False - if self.filter_type == "hibf" and self.hash_functions == 0: print_log("--filter-type hibf requires --hash-function value between 1 and 5") return False diff --git a/src/ganon/tax_util.py b/src/ganon/tax_util.py index 23ff406d..7173c187 100644 --- a/src/ganon/tax_util.py +++ b/src/ganon/tax_util.py @@ -3,27 +3,46 @@ import re import os import gzip -import tarfile from ganon.util import download, run, print_log, rm_files from io import StringIO -def parse_sequence_accession(input_files, info): +def parse_sequence_accession(input_files, info_cols, build_output_folder): """ - Look for sequence accession (anything from > to the first space) in all input files + Parse sequence accession (anything from > to the first space) in all input files + Split files by sequence and write them in subfolders """ + info = pd.DataFrame(columns=info_cols) + + # Create subfolders to distribute files + n_folders = 10 + run("seq 0 {n_folders} | xargs -i mkdir -p \"{build_output_folder}/{{}}\"".format(n_folders=n_folders-1, build_output_folder=build_output_folder), shell=True) + + # Randomly distribute sequences into subfolders + # cat | zcat -> compability with osx for file in input_files: - # cat | zcat -> compability with osx - run_cat = "cat {0} {1} | grep -o '^>[^ ]*' | sed 's/>//'".format(file, "| zcat" if file.endswith(".gz") else "") + run_cat = """ + cat {file} {zcat} | + awk '/^>/{{ + id=substr($1,2); + subf=int({n_folders} * rand())"/"; + file=(\"{build_output_folder}\" subf id \".fna\"); + print id\"\t\"file; + }} + {{print $0 > file}}' + """.format(file=file, + zcat="| zcat" if file.endswith(".gz") else "", + build_output_folder=build_output_folder, + n_folders=n_folders) + stdout = run(run_cat, shell=True, ret_stdout=True) - tmp_info = pd.read_csv(StringIO(stdout), header=None, names=['target']) - tmp_info["file"] = file + tmp_info = pd.read_csv(StringIO(stdout), header=None, names=['target', 'file'], delimiter="\t") info = pd.concat([info, tmp_info]) - return info + return info -def parse_file_accession(input_files, info): +def parse_file_accession(input_files, info_cols): """ Look for genbank/refseq assembly accession* pattern in the filename if not found, return basename of the file as target @@ -31,12 +50,15 @@ def parse_file_accession(input_files, info): *https://support.nlm.nih.gov/knowledgebase/article/KA-03451/en-us *https://https.ncbi.nlm.nih.gov/datasets/docs/v1/reference-docs/gca-and-gcf-explained/ """ + info = pd.DataFrame(columns=info_cols) + assembly_accessions = [] assembly_accession_pattern = re.compile("GC[A|F]_[0-9]+\.[0-9]+") for file in input_files: match = assembly_accession_pattern.search(file) assembly_accessions.append((match.group() if match else os.path.basename(file), file)) info[["target", "file"]] = pd.DataFrame(assembly_accessions) + return info def parse_genome_size_files(cfg, build_output_folder): @@ -70,10 +92,9 @@ def parse_genome_size_files(cfg, build_output_folder): fields = line.rstrip().split("\t") leaves_sizes[fields[0]] = int(fields[3]) - elif cfg.taxonomy == "gtdb": for file in files: - with gzip.open(file, "rt") as f: + with gzip.open(file, "rt") as f: # skip first line wiht header # col 0: accession (with GC_ RF_ prefix), col 13: genome_size, col 16: gtdb_taxonomy (d__Archaea;p__Thermoproteota;...) next(f) @@ -93,6 +114,7 @@ def parse_genome_size_files(cfg, build_output_folder): return leaves_sizes + def parse_genome_size_tax(tax_files): """ Parse last column of a .tax file and retrieve genome_sizes to a dict {node: size} @@ -110,6 +132,7 @@ def parse_genome_size_tax(tax_files): genome_sizes[node] = gsize return genome_sizes + def get_genome_size(cfg, nodes, tax, build_output_folder): """ Estimate genome sizes based on auxiliary files @@ -317,27 +340,36 @@ def get_sequence_info(cfg, info, tax, build_output_folder): def parse_acc2txid(info, acc2txid_files): count_acc2txid = {} unique_acc = set(info.index) - for acc2txid in acc2txid_files: - tmp_acc_node = pd.read_csv(acc2txid, - sep="\t", - header=None, - skiprows=1, - usecols=[1, 2], - names=["target", "node"], - index_col="target", - converters={"target": lambda x: x if x in unique_acc else None, "node": str}) - tmp_acc_node = tmp_acc_node[tmp_acc_node.index.notnull()] # keep only seqids used - tmp_acc_node = tmp_acc_node[tmp_acc_node["node"] != "0"] # filter out taxid==0 - # save count to return - count_acc2txid[acc2txid] = tmp_acc_node.shape[0] - # merge node(taxid) retrieved based on target(accesion) - if count_acc2txid[acc2txid]: - info.update(tmp_acc_node) - del tmp_acc_node - #if already found all seqids no need to parse all files till the end) - if sum(count_acc2txid.values()) == len(unique_acc): - break + # Parse very large acc2txid files in chunks to limit memory usage + chunksize = 10 ** 6 + for acc2txid in acc2txid_files: + count_acc2txid[acc2txid] = 0 + with pd.read_csv(acc2txid, + sep="\t", + header=None, + skiprows=1, + usecols=[1, 2], + names=["target", "node"], + index_col="target", + converters={"target": lambda x: x if x in unique_acc else None, "node": str}, + chunksize=chunksize) as reader: + + for tmp_acc_node in reader: + tmp_acc_node = tmp_acc_node[tmp_acc_node.index.notnull()] # keep only seqids used + tmp_acc_node = tmp_acc_node[tmp_acc_node["node"] != "0"] # filter out taxid==0 + + # If there was any match + if tmp_acc_node.shape[0]: + # merge node(taxid) retrieved based on target(accesion) + info.update(tmp_acc_node) + + # save count to return + count_acc2txid[acc2txid] += tmp_acc_node.shape[0] + + #if already found all seqids no need to parse all files till the end) + if sum(count_acc2txid.values()) == len(unique_acc): + break return count_acc2txid @@ -414,11 +446,13 @@ def run_eutils(cfg, info, build_output_folder, skip_taxid: bool=False, level: st # (-e) get taxid length # (-a) get assembly accession # (-m) get assembly name - run_get_seq_info_cmd = "{0} -i {1} -k {2} {3}".format(cfg.path_exec["get_seq_info"], - accessions_file, - "" if skip_taxid else "-e", - "-a -m" if level == "assembly" else "") - stdout = run(run_get_seq_info_cmd, ret_stdout=True, quiet=cfg.quiet) + # || true to ignore exit status in case some sequences were not retrieved + run_get_seq_info_cmd = "{0} -i {1} -k {2} {3} || true".format(cfg.path_exec["get_seq_info"], + accessions_file, + "" if skip_taxid else "-e", + "-a -m" if level == "assembly" else "") + + stdout = run(run_get_seq_info_cmd, ret_stdout=True, shell=True, quiet=cfg.quiet) # set "na" as NaN with na_values="na" if level == "assembly": diff --git a/src/ganon/util.py b/src/ganon/util.py index 214cfd00..b930d681 100644 --- a/src/ganon/util.py +++ b/src/ganon/util.py @@ -72,7 +72,7 @@ def validate_input_files(input_files_folder, input_extension, quiet, input_recur valid_input_files.add(i) elif os.path.isdir(i): if not input_extension: - print_log("--input-extension is required when using folders in the --input. Skipping: " + i, quiet) + print_log(" - --input-extension is required when using folders in the --input. Skipping: " + i, quiet) continue files_in_dir = 0 @@ -90,13 +90,13 @@ def validate_input_files(input_files_folder, input_extension, quiet, input_recur files_in_dir += 1 valid_input_files.add(f) - print_log(str(files_in_dir) + " valid file(s) [--input-extension " + input_extension + + print_log(" - " + str(files_in_dir) + " valid file(s) [--input-extension " + input_extension + (", --input-recursive" if input_recursive else "") + "] found in " + i, quiet) else: - print_log("Skipping invalid file/folder: " + i, quiet) + print_log(" - skipping invalid file/folder: " + i, quiet) - print_log("Total valid files: " + str(len(valid_input_files)) + "\n", quiet) + print_log(" - total valid files: " + str(len(valid_input_files)) + "", quiet) return valid_input_files diff --git a/tests/aux/Aux.hpp b/tests/aux/Aux.hpp index 1c243189..6d0ae33e 100644 --- a/tests/aux/Aux.hpp +++ b/tests/aux/Aux.hpp @@ -209,11 +209,7 @@ struct SeqTarget for ( uint16_t i = 0; i < files.size(); ++i ) { output_file << files[i]; - if ( sequence_as_target ) - { - output_file << '\t' << headers[i] << '\t' << headers[i]; - } - else if ( custom_targets ) + if ( custom_targets ) { output_file << '\t' << targets[i]; } @@ -237,8 +233,7 @@ struct SeqTarget std::vector< std::string > headers; std::vector< seqan3::dna4_vector > sequences; std::vector< std::string > targets; - bool sequence_as_target = false; - bool custom_targets = false; + bool custom_targets = false; }; } // namespace aux diff --git a/tests/ganon-build/GanonBuild.test.cpp b/tests/ganon-build/GanonBuild.test.cpp index 66bd8fa0..395bdd08 100644 --- a/tests/ganon-build/GanonBuild.test.cpp +++ b/tests/ganon-build/GanonBuild.test.cpp @@ -83,11 +83,7 @@ void validate_elements( const GanonBuild::Config cfg, const aux::SeqTarget& seqt auto counts_seq = agent.bulk_count( hashes ); // get bins from target of this sequence - std::vector< uint64_t > target_bins; - if ( seqtarget.sequence_as_target ) - target_bins = targets[seqtarget.headers[i]]; - else - target_bins = targets[seqtarget.targets[i]]; + const std::vector< uint64_t > target_bins = targets[seqtarget.targets[i]]; // sum matches for all bins of the targets this sequences belongs uint16_t expected_counts = 0; @@ -201,38 +197,6 @@ SCENARIO( "building indices", "[ganon-build]" ) config_build::validate_filter( cfg ); config_build::validate_elements( cfg, seqtarget ); } - - SECTION( "three columns - custom target" ) - { - std::string prefix{ folder_prefix + "input_file_three_cols" }; - auto cfg = config_build::defaultConfig( prefix ); - - std::vector< std::string > targets{ "T1", "T9", "T1", "T8", "T1", "T1", "T1", "T1", "T4", "T1" }; - - auto seqtarget = aux::SeqTarget( prefix, seqs, targets ); - seqtarget.sequence_as_target = false; - seqtarget.write_input_file( cfg.input_file ); - seqtarget.write_sequences_files(); - - REQUIRE( GanonBuild::run( cfg ) ); - config_build::validate_filter( cfg ); - config_build::validate_elements( cfg, seqtarget ); - } - - SECTION( "three columns - sequence as target" ) - { - std::string prefix{ folder_prefix + "input_file_three_cols_sequence_target" }; - auto cfg = config_build::defaultConfig( prefix ); - - auto seqtarget = aux::SeqTarget( prefix, seqs ); - seqtarget.sequence_as_target = true; - seqtarget.write_input_file( cfg.input_file ); - seqtarget.write_sequences_files(); - - REQUIRE( GanonBuild::run( cfg ) ); - config_build::validate_filter( cfg ); - config_build::validate_elements( cfg, seqtarget ); - } } SECTION( "--max-fp" ) diff --git a/tests/ganon-build/data/SEQ0.fna.gz b/tests/ganon-build/data/SEQ0.fna.gz new file mode 100644 index 0000000000000000000000000000000000000000..55cd455c8406a8f7f88d50c827167065a563b9c5 GIT binary patch literal 12852 zcmV-4GRw^$iwFpPj>BaD15-s&FfL|pVE|Q{QH~``Oho@#4J$yhEaY#5_rDgVd`{$T z5b$Pt`qr(=blMRo>c9T?zyIey|D$HMdglJSUDaE&-Sh8H_qX}?->&c9|NUoY|NZLs z&;NdP|NVKl=ijIQ{aUF7?ve}DaVfq&Qi{`&oW|9$$qet#F7{(f`rYIm*ruEdZ3 zeW>n2|9<@MU-!F%-S4CSe&yf2?00|P&;I@Pd^gnpZsEJ{{VrVJ-~Ro^cf-q#d{>;i 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zQq*@@W@2@Rw9aZ=N?plKPwu|l0Tq%95K)#B%6W&}v^r!^=+@qDVDsB~vwS22Z#`*y zzcu7Vi8W_}>Ly%2y;1Sh>a#=`scs8mcJ`ZXcsfmcR5unEqVZ;f7a;iJ-u>FHA$G7LLSjOtWhhcY zf(@La48)r!?)D_949fE)bYysYcI~v(!zP&N&&->j@LL&Zh7BbQ5HEse;DNDBNNc2q zFvPyTLN>FzHOioKlQ{w7X@9)fCioN!Cw#+hBLa^BRalk%k100: + if rank in Config.choices_default_ranks and val > 100: print("Inconsistent percentage by rank (>100%)") return None # Check if counts are consistent if ((res["tre_pd"]["unique"] + res["tre_pd"]["shared"] + res["tre_pd"]["children"]) > res["tre_pd"]["cumulative"]).any(): print("Inconsistent counts") - return None + return None # Check if percentage/abundance is consistent # nodes cannot have higher percentage than parents - target_perc = dict(zip(res["tre_pd"]["target"], res["tre_pd"]["cumulative_perc"])) + target_perc = dict( + zip(res["tre_pd"]["target"], res["tre_pd"]["cumulative_perc"])) for l in res["tre_pd"]["lineage"]: # skip empty nodes (e.g 241||412412 -> 241|412412) lineage = [n for n in l.split("|") if n] @@ -342,11 +349,12 @@ def report_sanity_check_and_parse(params, sum_full_percentage: bool=True): multi_res[out_tre] = res # If only one output, return directly - if len(multi_res)==1: + if len(multi_res) == 1: return res else: return multi_res + def table_sanity_check_and_parse(params): # Provide sanity checks for outputs (not specific to a test) and return loaded data @@ -358,29 +366,29 @@ def table_sanity_check_and_parse(params): res["out_pd"] = parse_tsv(params["output_file"]) # check if all values are positive - if not (res["out_pd"].values>=0 ).all(): + if not (res["out_pd"].values >= 0).all(): print("Invalid negative value") return None # specific for percentage output - if params["output_value"]=="percentage": + if params["output_value"] == "percentage": # check if all percentages are on the 0-1 range - if not (res["out_pd"].values<=1).all(): + if not (res["out_pd"].values <= 1).all(): print("Invalid table percentage (>1)") return None # Check if sum of the lines do not pass 1 - if not (res["out_pd"].sum(axis=1)<=1).all(): + if not (res["out_pd"].sum(axis=1) <= 1).all(): print("Invalid line value (>1)") return None - return res + def download_bulk_files(download_dir): - bulk_files = {"https://ftp.ncbi.nlm.nih.gov/": - ["pub/taxonomy/accession2taxid/dead_nucl.accession2taxid.gz", + bulk_files = {"https://ftp.ncbi.nlm.nih.gov/": + ["pub/taxonomy/accession2taxid/dead_nucl.accession2taxid.gz", "pub/taxonomy/accession2taxid/nucl_gb.accession2taxid.gz", "genomes/refseq/assembly_summary_refseq.txt", "genomes/refseq/assembly_summary_refseq_historical.txt", @@ -389,7 +397,7 @@ def download_bulk_files(download_dir): "genomes/ASSEMBLY_REPORTS/species_genome_size.txt.gz"], "https://data.gtdb.ecogenomic.org/releases/latest/": ["ar53_metadata.tsv.gz", - "bac120_metadata.tsv.gz"] + "bac120_metadata.tsv.gz"] } for url, files in bulk_files.items(): @@ -398,6 +406,50 @@ def download_bulk_files(download_dir): print("File found: " + download_dir + file) else: p = os.path.dirname(file) - print("Downloading " + url + file + " -> " + download_dir + p + file) + print("Downloading " + url + file + + " -> " + download_dir + p + file) os.makedirs(download_dir + p, exist_ok=True) - download([url + file], download_dir + p + "/") \ No newline at end of file + download([url + file], download_dir + p + "/") + + +def write_input_file(files, + assembly_summary, + output_file, + cols: list = ["file", "target", "node", "specialization", "specialization_name"], + sequence: bool = False): + """ + Generates a simulated --input-file based on an assembly summary and a list of files + """ + + input_file = {} + for file in files: + input_file[Path(file).name] = {"file": file} + + with open(assembly_summary, "r") as asf: + for line in asf: + fields = line.split("\t") + # filename from url + file = Path(fields[19]).name + "_genomic.fna.gz" + + if file not in input_file: + continue + + # assembly accession as target + target = fields[0] + # taxid as node + node = fields[5] + # use biosample as specialization + spec = fields[2] + # use name + strain for specialization name + spec_name = fields[7] + " " + fields[8] + + input_file[file].update({"target": target, "node": node, "specialization": spec, "specialization_name": spec_name}) + + with open(output_file, "w") as outf: + for file, vals in input_file.items(): + if sequence: + for seq in list_sequences([vals["file"]]): + vals["target"] = seq + print("\t".join([v for k,v in vals.items() if k in cols]), file=outf) + else: + print("\t".join([v for k,v in vals.items() if k in cols]), file=outf) From 4a0044cfe26a010cffaec0e3df5225748ebb2b3c Mon Sep 17 00:00:00 2001 From: pirovc <4673375+pirovc@users.noreply.github.com> Date: Sun, 11 Feb 2024 13:46:33 +0100 Subject: [PATCH 3/5] Feature/ci test hibf (#284) * test compile raptor * attempt with conda * debug travis * debug travis * add tests hibf * add update --- .travis.yml | 27 ++++++++------- docs/index.md | 1 + tests/ganon/data/classify/test_db.hibf | Bin 475930 -> 0 bytes tests/ganon/data/classify/test_db.tax | 29 ---------------- tests/ganon/integration/test_build.py | 11 +++++-- tests/ganon/integration/test_build_custom.py | 11 +++++-- tests/ganon/integration/test_classify.py | 33 +++++++------------ tests/ganon/integration/test_update.py | 14 +++++--- 8 files changed, 54 insertions(+), 72 deletions(-) delete mode 100644 tests/ganon/data/classify/test_db.hibf delete mode 100644 tests/ganon/data/classify/test_db.tax diff --git a/.travis.yml b/.travis.yml index fafde3d4..bd29612e 100755 --- a/.travis.yml +++ b/.travis.yml @@ -7,20 +7,15 @@ matrix: os: linux addons: apt: - sources: - - ubuntu-toolchain-r-test packages: - g++-11 - parallel env: - MATRIX_EVAL="CC=gcc-11 && CXX=g++-11 && BUILD_TYPE=Release" - - name: "linux gcc-12" os: linux addons: apt: - sources: - - ubuntu-toolchain-r-test packages: - g++-12 - parallel @@ -31,8 +26,6 @@ matrix: os: linux addons: apt: - sources: - - ubuntu-toolchain-r-test packages: - g++-11 - parallel @@ -43,22 +36,34 @@ matrix: before_install: - eval "${MATRIX_EVAL}" - python3 -m pip install --upgrade pip + - wget https://repo.anaconda.com/miniconda/Miniconda3-latest-Linux-x86_64.sh -O miniconda.sh + - bash miniconda.sh -b -p $HOME/miniconda + - source "$HOME/miniconda/etc/profile.d/conda.sh" + - conda config --set always_yes yes --set changeps1 no + - conda update -q conda + - conda info -a + - conda activate base + +install: - python3 -m pip install "pandas>=1.2.0" - python3 -m pip install "multitax>=1.3.1" + - python3 -m pip install "parameterized>=0.9.0" - if [ "$BUILD_TYPE" == "Coverage" ]; then python3 -m pip install coverage; fi - -script: + - conda install -c bioconda -c conda-forge "raptor=3.0.1" - mkdir build && cd build - cmake -DCMAKE_BUILD_TYPE=$BUILD_TYPE -DVERBOSE_CONFIG=ON -DCMAKE_EXPORT_COMPILE_COMMANDS=ON -DCONDA=OFF .. - cat compile_commands.json - cmake --build . -- -j2 - ctest -VV . - sudo make install - - ganon-build -h # ret 0 - - ganon-classify -h # ret 0 - cd .. + +script: + - ganon-build --version + - ganon-classify --version + - raptor --version - if [ "$BUILD_TYPE" == "Coverage" ]; then python3 -m coverage run --omit="/usr/*,tests/*" -m unittest discover -s tests/ganon/integration/ -v; else diff --git a/docs/index.md b/docs/index.md index 59571515..8bcd24b6 100644 --- a/docs/index.md +++ b/docs/index.md @@ -131,6 +131,7 @@ ganon -h #### Running tests ```bash +python3 -m pip install "parameterized>=0.9.0" python3 -m unittest discover -s tests/ganon/integration/ python3 -m unittest discover -s tests/ganon/integration_online/ # optional - downloads large files cd build/ diff --git a/tests/ganon/data/classify/test_db.hibf b/tests/ganon/data/classify/test_db.hibf deleted file mode 100644 index 5755549ff8e87c1023c09c2c556a44389072c327..0000000000000000000000000000000000000000 GIT binary patch literal 0 HcmV?d00001 literal 475930 zcmbTee{^KkbuRdwbI-jxbyY1+l}hT85cpK-N|d?)_5XaBS z@>>h%$D8rYtjTP}&RTD+S?`ZGwHD}C)xGEJv-h{Z{q2351NaF2xBqJY*%$otKeznL 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dryophthoridicola 7882008 -1972133 1971485 forma specialis endosymbiont of Rhynchophorus ferrugineus 7882008 -2012515 1801617 species Candidatus Pacearchaeota archaeon ex4484_31 1970502 -2565781 2856051 species Candidatus Nanohalobium constans 6896757 -871271 884215 strain Candidatus Zinderia insecticola CARI 985251 -118884 1236 no rank Gammaproteobacteria incertae sedis 7882008 -1224 2 phylum Proteobacteria 3941004 -1236 1224 class Gammaproteobacteria 5418880 -131567 1 no rank cellular organisms 4222504 -1462430 1783276 phylum Candidatus Nanohaloarchaeota 6896757 -1783276 2157 clade DPANN group 4433629 -1801617 1783276 clade Candidatus Pacearchaeota 1970502 -2 131567 superkingdom Bacteria 3941004 -204619 118884 genus Candidatus Nardonella 7882008 -2157 131567 superkingdom Archaea 4433629 -28216 1224 class Betaproteobacteria 985251 -2856051 2856054 genus Candidatus Nanohalobium 6896757 -2856052 1462430 class Candidatus Nanohalobia 6896757 -2856053 2856052 order Candidatus Nanohalobiales 6896757 -2856054 2856053 family Candidatus Nanohalobiaceae 6896757 -75682 80840 family Oxalobacteraceae 985251 -80840 28216 order Burkholderiales 985251 -884214 75682 genus Candidatus Zinderia 985251 -884215 884214 species Candidatus Zinderia insecticola 985251 -GCF_004296495.1 1972133 assembly endosymbiont of Rhynchophorus ferrugineus 7882008 -GCA_000147015.1 871271 assembly Candidatus Zinderia insecticola CARI 985251 -GCF_009617975.1 2565781 assembly Candidatus Nanohalobium constans LC1Nh 6896757 -GCA_002254805.1 2012515 assembly Candidatus Pacearchaeota archaeon ex4484_31 1970502 diff --git a/tests/ganon/integration/test_build.py b/tests/ganon/integration/test_build.py index 1e5dcb14..752f1fe2 100644 --- a/tests/ganon/integration/test_build.py +++ b/tests/ganon/integration/test_build.py @@ -13,11 +13,14 @@ from utils import build_sanity_check_and_parse data_dir = base_dir + "data/" +from parameterized import parameterized_class +@parameterized_class([ + { "filter_type": "ibf"}, + { "filter_type": "hibf" }, +]) class TestBuild(unittest.TestCase): - results_dir = base_dir + "results/integration/build/" - default_params = {"organism_group": ["archaea", "bacteria", "viral"], "source": ["genbank"], "taxonomy": "skip", @@ -37,6 +40,10 @@ def setUpClass(self): # viral GCA_004132065.1 txid 2161879 # Export local_dir for genome_updater, uses local folder as repository os.environ["local_dir"] = os.path.abspath(data_dir + "build/") + + # parametrization for filter type + self.default_params["filter_type"] = self.filter_type + self.results_dir = base_dir + "results/integration/build_" + self.filter_type + "/" setup_dir(self.results_dir) def test_og_arc_bac_vir(self): diff --git a/tests/ganon/integration/test_build_custom.py b/tests/ganon/integration/test_build_custom.py index 9ea2a968..ef51d63d 100644 --- a/tests/ganon/integration/test_build_custom.py +++ b/tests/ganon/integration/test_build_custom.py @@ -15,15 +15,17 @@ from utils import write_input_file data_dir = base_dir + "data/" +from parameterized import parameterized_class +@parameterized_class([ + { "filter_type": "ibf"}, + { "filter_type": "hibf" }, +]) class TestBuildCustom(unittest.TestCase): - results_dir = base_dir + "results/integration/build-custom/" - default_params = {"input": data_dir + "build-custom/files/", "taxonomy": "skip", "threads": 1, - "filter_type": "ibf", "write_info_file": True, "keep_files": True, "verbose": True, @@ -31,6 +33,9 @@ class TestBuildCustom(unittest.TestCase): @classmethod def setUpClass(self): + # parametrization for filter type + self.default_params["filter_type"] = self.filter_type + self.results_dir = base_dir + "results/integration/build-custom_" + self.filter_type + "/" setup_dir(self.results_dir) def test_input_folder(self): diff --git a/tests/ganon/integration/test_classify.py b/tests/ganon/integration/test_classify.py index 29915389..79af2223 100644 --- a/tests/ganon/integration/test_classify.py +++ b/tests/ganon/integration/test_classify.py @@ -12,18 +12,23 @@ from utils import check_files data_dir = base_dir + "data/" +from parameterized import parameterized_class +@parameterized_class([ + { "filter_type": "ibf"}, + { "filter_type": "hibf" }, +]) class TestClassify(unittest.TestCase): - results_dir = base_dir + "results/integration/classify/" - default_params = {"db_prefix": [results_dir + "base_build"], - "paired_reads": [data_dir+"classify/sim.1.fq.gz", + default_params = {"paired_reads": [data_dir+"classify/sim.1.fq.gz", data_dir+"classify/sim.2.fq.gz"], "verbose": True, "quiet": False} @classmethod def setUpClass(self): + self.results_dir = base_dir + "results/integration/classify_" + self.filter_type + "/" + self.default_params["db_prefix"] = [self.results_dir + "base_build"] setup_dir(self.results_dir) # Build base database build_params = {"db_prefix": self.results_dir + "base_build", @@ -34,7 +39,7 @@ def setUpClass(self): "genome_size_file": data_dir + "build-custom/species_genome_size.txt.gz", "level": "assembly", "threads": 1, - "filter_type": "ibf", + "filter_type": self.filter_type, "keep_files": True, "write_info_file": True, "verbose": True, @@ -52,7 +57,7 @@ def setUpClass(self): "genome_size_file": data_dir + "build-custom/species_genome_size.txt.gz", "level": "assembly", "threads": 1, - "filter_type": "ibf", + "filter_type": self.filter_type, "keep_files": True, "write_info_file": True, "verbose": True, @@ -139,6 +144,7 @@ def test_multiple_matches_em(self): params["rel_cutoff"] = 0.001 params["rel_filter"] = 1 + print(params) # Build config from params cfg = Config("classify", **params) # Run @@ -195,23 +201,6 @@ def test_multiple_matches_skip(self): # There is no .one output self.assertFalse(check_files(params["output_prefix"], "one")) - - def test_hibf(self): - """ - Test ganon classify with HIBF - """ - params = self.default_params.copy() - params["db_prefix"] = data_dir + "classify/test_db" - params["output_prefix"] = self.results_dir + "hibf" - params["verbose"] = True - # Build config from params - cfg = Config("classify", **params) - # Run - self.assertTrue(run_ganon(cfg, params["output_prefix"]), "ganon classify exited with an error") - # General sanity check of results - res = classify_sanity_check_and_parse(vars(cfg)) - self.assertIsNotNone(res, "ganon table has inconsistent results") - def test_report_params(self): """ Test ganon classify with report parameters diff --git a/tests/ganon/integration/test_update.py b/tests/ganon/integration/test_update.py index 38c036fa..b52f3819 100644 --- a/tests/ganon/integration/test_update.py +++ b/tests/ganon/integration/test_update.py @@ -14,13 +14,17 @@ from utils import build_sanity_check_and_parse data_dir = base_dir + "data/" +from parameterized import parameterized_class +@parameterized_class([ + { "filter_type": "ibf"}, + { "filter_type": "hibf" }, +]) class TestUpdate(unittest.TestCase): - results_dir = base_dir + "results/integration/update/" - @classmethod - def setUpClass(self): + def setUpClass(self): + self.results_dir = base_dir + "results/integration/update_" + self.filter_type + "/" # 3 small genomes from genbank (also on GTDB R207 for bac arc) # archaea GCA_002254805.1 # bacteria GCA_000147015.1 @@ -40,7 +44,7 @@ def test_output_db_prefix(self): "taxonomy": "skip", "level": "assembly", "threads": 1, - "filter_type": "ibf", + "filter_type": self.filter_type, "write_info_file": True, "keep_files": True, "verbose": True, @@ -90,7 +94,7 @@ def test_db_prefix(self): "taxonomy": "skip", "level": "assembly", "threads": 1, - "filter_type": "ibf", + "filter_type": self.filter_type, "write_info_file": True, "keep_files": True, "verbose": True, From c2528d4f711b3fd56ee6bed3d61c12263e832a3e Mon Sep 17 00:00:00 2001 From: "Vitor C. Piro" Date: Sun, 11 Feb 2024 14:18:01 +0100 Subject: [PATCH 4/5] docs, config --- docs/custom_databases.md | 4 +-- docs/index.md | 31 ++++++++++++------------ src/ganon/config.py | 2 +- tests/ganon/integration/test_classify.py | 1 - 4 files changed, 18 insertions(+), 20 deletions(-) diff --git a/docs/custom_databases.md b/docs/custom_databases.md index 27e8def3..7edcc9d5 100644 --- a/docs/custom_databases.md +++ b/docs/custom_databases.md @@ -68,8 +68,8 @@ The classification max. level against this database will depend on the value set #### Files, taxonomy and specialization ``` -sequences.fasta FILE_A 562 ID44444 Escherichia coli TW10119 -others.fasta FILE_B 623 ID55555 Shigella flexneri 1a +sequences.fasta sequences 562 ID44444 Escherichia coli TW10119 +others.fasta others 623 ID55555 Shigella flexneri 1a ``` The classification max. level against this database will depend on the value set for `--level`: diff --git a/docs/index.md b/docs/index.md index 8bcd24b6..ce0dc205 100644 --- a/docs/index.md +++ b/docs/index.md @@ -49,8 +49,10 @@ python3 -V # Install packages via pip or conda: # PIP python3 -m pip install "pandas>=1.2.0" "multitax>=1.3.1" +wget --quiet --show-progress https://raw.githubusercontent.com/pirovc/genome_updater/master/genome_updater.sh && chmod +x genome_updater.sh + # Conda/Mamba (alternative) -conda install "pandas>=1.2.0" "multitax>=1.3.1" +conda install -c bioconda -c conda-forge "pandas>=1.2.0" "multitax>=1.3.1" "genome_updater>=0.6.3" ``` ### C++ dependencies @@ -90,7 +92,7 @@ sudo make install # optional ### Installing raptor -The easiest way to install [raptor](https://github.com/seqan/raptor) is via conda with `conda install -c bioconda -c conda-forge "raptor>=3.0.1"` (already included in ganon install via conda). +The easiest way to install [raptor](https://github.com/seqan/raptor) is via conda with `conda install -c bioconda -c conda-forge "raptor=3.0.1"` (already included in ganon install via conda). !!! Note raptor is required to build databases with the Hierarchical Interleaved Bloom Filter (`ganon build --filter-type hibf`) @@ -131,7 +133,7 @@ ganon -h #### Running tests ```bash -python3 -m pip install "parameterized>=0.9.0" +python3 -m pip install "parameterized>=0.9.0" # Alternative: conda install -c conda-forge "parameterized>=0.9.0" python3 -m unittest discover -s tests/ganon/integration/ python3 -m unittest discover -s tests/ganon/integration_online/ # optional - downloads large files cd build/ @@ -147,7 +149,7 @@ usage: ganon [-h] [-v] - - - - - - - - - - _ _ _ _ _ (_|(_|| |(_)| | - _| v. 2.0.1 + _| v. 2.1.0 - - - - - - - - - - positional arguments: @@ -271,18 +273,15 @@ required arguments: Database output prefix (default: None) custom arguments: - -n , --input-file Manually set information for input files: file [target node specialization - specialization name]. target is the sequence identifier if --input-target sequence (file - can be repeated for multiple sequences). if --input-target file and target is not set, filename - is used. node is the taxonomic identifier. Mutually exclusive --input (default: None) - -a , --input-target Target to use [file, sequence]. By default: 'file' if multiple input files are provided or - --input-file is set, 'sequence' if a single file is provided. Using 'file' is recommended and - will speed-up the building process (default: None) - -l , --level Use a specialized target to build the database. By default, --level is the --input-target. - Options: any available taxonomic rank [species, genus, ...] or 'leaves' (requires --taxonomy). - Further specialization options [assembly, custom]. assembly will retrieve and use the assembly - accession and name. custom requires and uses the specialization field in the --input-file. - (default: None) + -n , --input-file Tab-separated file with all necessary file/sequence information. Fields: file [ target + node specialization specialization name]. For details: + https://pirovc.github.io/ganon/custom_databases/. Mutually exclusive --input (default: None) + -a , --input-target Target to use [file, sequence]. Parse input by file or by sequence. Using 'file' is recommended + and will speed-up the building process (default: file) + -l , --level Max. level to build the database. By default, --level is the --input-target. Options: any + available taxonomic rank [species, genus, ...] or 'leaves' (requires --taxonomy). Further + specialization options [assembly, custom]. assembly will retrieve and use the assembly accession + and name. custom requires and uses the specialization field in the --input-file. (default: None) -m [ ...], --taxonomy-files [ ...] Specific files for taxonomy - otherwise files will be downloaded (default: None) -z [ ...], --genome-size-files [ ...] diff --git a/src/ganon/config.py b/src/ganon/config.py index 0c3f0198..fdd43e2b 100644 --- a/src/ganon/config.py +++ b/src/ganon/config.py @@ -88,7 +88,7 @@ def __init__(self, which: str=None, **kwargs): build_custom_required_args.add_argument("-c", "--input-recursive", action="store_true", help="Look for files recursively in folder(s) provided with --input") build_custom_args = build_custom_parser.add_argument_group("custom arguments") - build_custom_args.add_argument("-n", "--input-file", type=file_exists, metavar="", help="Manually set information for input files: file [target node specialization specialization name]. target is the sequence identifier if --input-target sequence (file can be repeated for multiple sequences). if --input-target file and target is not set, filename is used. node is the taxonomic identifier. Mutually exclusive --input") + build_custom_args.add_argument("-n", "--input-file", type=file_exists, metavar="", help="Tab-separated file with all necessary file/sequence information. Fields: file [ target node specialization specialization name]. For details: https://pirovc.github.io/ganon/custom_databases/. Mutually exclusive --input") build_custom_args.add_argument("-a", "--input-target", type=str, default="file", metavar="", help="Target to use [file, sequence]. Parse input by file or by sequence. Using 'file' is recommended and will speed-up the building process", choices=self.choices_input_target) build_custom_args.add_argument("-l", "--level", type=str, metavar="", help="Max. level to build the database. By default, --level is the --input-target. Options: any available taxonomic rank [species, genus, ...] or 'leaves' (requires --taxonomy). Further specialization options [" + ", ".join(self.choices_level) + "]. assembly will retrieve and use the assembly accession and name. custom requires and uses the specialization field in the --input-file.") build_custom_args.add_argument("-m", "--taxonomy-files", type=file_exists, nargs="*", metavar="", help="Specific files for taxonomy - otherwise files will be downloaded") diff --git a/tests/ganon/integration/test_classify.py b/tests/ganon/integration/test_classify.py index 79af2223..00aae667 100644 --- a/tests/ganon/integration/test_classify.py +++ b/tests/ganon/integration/test_classify.py @@ -144,7 +144,6 @@ def test_multiple_matches_em(self): params["rel_cutoff"] = 0.001 params["rel_filter"] = 1 - print(params) # Build config from params cfg = Config("classify", **params) # Run From 1dfd757bdb352b87e5060b0b3bd7e096a5e3a8cf Mon Sep 17 00:00:00 2001 From: "Vitor C. Piro" Date: Sun, 11 Feb 2024 15:42:18 +0100 Subject: [PATCH 5/5] docs --- docs/index.md | 3 ++- 1 file changed, 2 insertions(+), 1 deletion(-) diff --git a/docs/index.md b/docs/index.md index ce0dc205..8449b4b6 100644 --- a/docs/index.md +++ b/docs/index.md @@ -41,6 +41,7 @@ However, there are possible performance benefits compiling ganon from source in - python >=3.6 - pandas >=1.2.0 - [multitax](https://github.com/pirovc/multitax) >=1.3.1 +- [genome_updater](https://github.com/pirovc/genome_updater) >=0.6.3 ```bash # Python version should be >=3.6 @@ -60,7 +61,7 @@ conda install -c bioconda -c conda-forge "pandas>=1.2.0" "multitax>=1.3.1" "geno - CMake >=3.4 - zlib - bzip2 -- raptor >=3.0.1 +- raptor ==3.0.1 !!! tip If your system has GCC version 10 or below, you can create an environment with the latest conda-forge GCC version and dependencies: `conda create -c conda-forge -n gcc-conda gcc gxx zlib bzip2 cmake` and activate the environment with: `source activate gcc-conda`.

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