# Using sourmash from the command line ```{contents} Contents :depth: 3 ``` From the command line, sourmash can be used to create [FracMinHash sketches][0] from DNA and protein sequences, compare them to each other, and plot the results; these sketches are saved into "signature files". These signatures allow you to estimate sequence similarity and containment quickly and accurately in large collections, among other capabilities. sourmash also provides a suite of metagenome functionality. This includes genome search in metagenomes, metagenome decomposition into a list of genomes from a database, and taxonomic classification functionality. The sourmash team provides a collection of prepared [databases](databases.md) for GTDB and GenBank. There is an increasingly large ecosystem of plugins that support [high-performance search and sketching](https://github.com/sourmash-bio/sourmash_plugin_branchwater), [more advanced plotting capabilities](https://github.com/sourmash-bio/sourmash_plugin_betterplot/), and [streaming sketching of large collections of genomes](https://github.com/sourmash-bio/sourmash_plugin_directsketch). Please see the [mash software][1] and the [mash paper (Ondov et al., 2016)][2] for background information on how and why MinHash sketches work. The [FracMinHash preprint (Irber et al, 2022)](https://www.biorxiv.org/content/10.1101/2022.01.11.475838) describes FracMinHash sketches as well as the metagenome-focused features of sourmash. sourmash uses a subcommand syntax, so all commands start with `sourmash` followed by a subcommand specifying the action to be taken. ## An example Download three bacterial genomes from NCBI: ``` curl -L -O https://ftp.ncbi.nlm.nih.gov/genomes/all/GCF/000/017/325/GCF_000017325.1_ASM1732v1/GCF_000017325.1_ASM1732v1_genomic.fna.gz curl -L -O https://ftp.ncbi.nlm.nih.gov/genomes/all/GCF/000/021/665/GCF_000021665.1_ASM2166v1/GCF_000021665.1_ASM2166v1_genomic.fna.gz curl -L -O https://ftp.ncbi.nlm.nih.gov/genomes/refseq/bacteria/Escherichia_coli/reference/GCF_000005845.2_ASM584v2/GCF_000005845.2_ASM584v2_genomic.fna.gz ``` Compute sourmash signatures for them all: ``` sourmash sketch dna -p k=31 *.fna.gz ``` This will produce three `.sig` files containing MinHash signatures using a k-mer size of 31. Next, compare all the signatures to each other: ``` sourmash compare *.sig -o cmp.dist ``` Finally, plot a dendrogram: ``` sourmash plot cmp.dist --labels ``` This will output three files, `cmp.dist.dendro.png`, `cmp.dist.matrix.png`, and `cmp.dist.hist.png`, containing a clustering & dendrogram of the sequences, a similarity matrix and heatmap, and a histogram of the pairwise similarities between the three genomes. Matrix: ![Matrix](_static/cmp.matrix.png) Here, the two genomes that cluster together are strains of the same species, while the third is from a completely different genus. ## The `sourmash` command and its subcommands To get a list of subcommands, run `sourmash` without any arguments. Please use the command line option `--help` to get more detailed usage information for each command. All signature saving commands can save to a variety of formats (we suggest `.zip` files) and all signature loading commands can load signatures from any of these formats. There are seven main subcommands: `sketch`, `compare`, `plot`, `search`, `gather`, `index`, and `prefetch`. See [the tutorial](tutorials.md) for a walkthrough of these commands. * `sketch` creates signatures. * `compare` compares signatures and builds a similarity matrix. * `plot` plots similarity matrices created by `compare`. * `search` finds matches to a query signature in a collection of signatures. * `gather` finds the best reference genomes for a metagenome, using the provided collection of signatures. * `index` builds fast indexes for searching many (thousands to millions) of signatures. * `prefetch` selects signatures of interest from a very large collection of signatures, for later processing. There are also a number of commands that work with taxonomic information; these are grouped under the `sourmash tax` and `sourmash lca` subcommands. `sourmash tax` commands: * `tax metagenome` - summarize metagenome gather results at each taxonomic rank. * `tax genome` - summarize single-genome gather results and report most likely classification. * `tax annotate` - annotate gather results with lineage information (no summarization or classification). * `tax prepare` - prepare and/or combine taxonomy files. * `tax grep` - subset taxonomies and create picklists based on taxonomy string matches. * `tax summarize` - print summary information (counts of lineages) for a taxonomy lineages file or database. `sourmash lca` commands: ```{attention} We do not recommend using the `lca` subcommands for taxonomic analysis any more; please use `sourmash tax` instead. See [taxonomic profiling with sourmash](classifying-signatures.md#taxonomic-profiling-with-sourmash) for more information. ``` * `lca classify` classifies many signatures against an LCA database. * `lca summarize` summarizes the content of metagenomes using an LCA database. * `lca index` creates a database for use with LCA subcommands. * `lca rankinfo` summarizes the content of a database. * `lca compare_csv` compares lineage spreadsheets, e.g. those output by `lca classify`. See [the LCA tutorial](tutorials-lca.md) for a walkthrough of some of these commands. Finally, there are a number of utility and information commands: * `info` shows version and software information. * `sbt_combine` combines multiple SBT indexes. * `categorize` is an experimental command to categorize many signatures. * `watch` is an experimental command to classify a stream of sequencing data. * `multigather` is an experimental command to run multiple gathers against the same collection of databases. Please use the command line option `--help` to get more detailed usage information for each command. ### `sourmash sketch` - make sourmash signatures from sequence data Most of the commands in sourmash work with **signatures**, which contain information about genomic or proteomic sequences. Each signature contains one or more **sketches**, which are compressed versions of these sequences. Using sourmash, you can search, compare, and analyze these sequences in various ways. To create a signature with one or more sketches, you use the `sourmash sketch` command. There are four main commands: ``` sourmash sketch dna sourmash sketch protein sourmash sketch translate sourmash sketch fromfile ``` The `sketch dna` command reads in **DNA sequences** and outputs **DNA sketches**. The `sketch protein` command reads in **protein sequences** and outputs **protein sketches**. The `sketch translate` command reads in **DNA sequences**, translates them in all six frames, and outputs **protein sketches**. The `sketch fromfile` command takes in a CSV file containing the locations of genomes and proteomes, and outputs all of the requested sketches. It is primarily intended for large-scale database construction. (`fromfile` is a new command as of sourmash v4.4.0.) All of the `sourmash sketch` commands take FASTA or FASTQ sequences as input; input data can be uncompressed, compressed with gzip, or compressed with bzip2. The output will be one or more signature files that can be used by other sourmash commands. Please see [the `sourmash sketch` documentation page](sourmash-sketch.md) for details on `sketch`, and see [Using sourmash: a practical guide](using-sourmash-a-guide.md) for more information on creating signatures. ### `sourmash compute` - make sourmash signatures from sequence data **Note: `sourmash compute` is deprecated in sourmash 4.0 and will be removed in sourmash 5.0; please switch to using `sourmash sketch`, above.** The `compute` subcommand computes and saves signatures for each sequence in one or more sequence files. It takes as input FASTA or FASTQ files, and these files can be uncompressed or compressed with gzip or bzip2. The output will be one or more JSON signature files that can be used with `sourmash compare`. Please see [Using sourmash: a practical guide](using-sourmash-a-guide.md) for more information on computing signatures. ______ Usage: ``` sourmash compute [ ... ] ``` Optional arguments: ``` --ksizes K1[,K2,K3] -- one or more k-mer sizes to use; default is 31 --force -- recompute existing signatures; convert non-DNA characters to N --output -- save all the signatures to this file; can be '-' for stdout. --track-abundance -- compute and save k-mer abundances. --name-from-first -- name the signature based on the first sequence in the file --singleton -- instead of computing a single signature for each input file, compute one for each sequence --merged -- compute a single signature for all of the input files, naming it ``` ### `sourmash compare` - compare many signatures **Note:** As of 2025, we have a much faster implementation of `compare` called `multisearch` available in [the branchwater plugin](https://github.com/sourmash-bio/sourmash_plugin_branchwater). It is multithreaded and much more memory efficient than `compare`, although it does accept a slightly more restricted set of inputs. The `compare` subcommand compares one or more signatures (created with `sketch`) using estimated [Jaccard index][3] or (if signatures are created with `-p abund`) the [angular similarity](https://en.wikipedia.org/wiki/Cosine_similarity#Angular_distance_and_similarity). Jaccard is most appropriate for genome comparisons, while angular similarity has been used for comparing metagenomes (e.g. see [simka](https://github.com/GATB/simka)). Use `--ignore-abundance` to force Jaccard. The default output of `compare` is a text display of a similarity matrix where each entry `[i, j]` contains the estimated Jaccard index between input signature `i` and input signature `j`. The output matrix can be saved to a numpy binary file with `--output ` and used with the `sourmash plot` subcommand (or loaded with `numpy.load(...)`. Using `--csv ` will output a CSV file that can be loaded into other languages than Python, such as R. As of sourmash 4.4.0, `compare` also supports Average Nucleotide Identity (ANI) estimates instead of Jaccard or containment index; use `--ani` to enable this. Usage: ``` sourmash compare [ ... ] ``` Options: * `--output ` -- save the output matrix to this file, as a numpy binary matrix. * `--distance-matrix` -- create and output a distance matrix, instead of a similarity matrix. * `--ksize ` -- select sketches at this k-mer size * `--containment` -- calculate containment instead of similarity; `C(i, j) = size(i intersection j) / size(i)` * `--ani` -- output estimates of Average Nucleotide Identity (ANI) instead of Jaccard similarity or containment. * `--from-file ` -- append the list of files in this text file to the input signatures. * `--ignore-abundance` -- ignore abundances in signatures and calculate Jaccard instead of angular similarity. * `--picklist ::` -- select a subset of signatures with [a picklist](#using-picklists-to-subset-large-collections-of-signatures) * `--csv ` -- save the output matrix in CSV format. * `--labels-to ` -- create a CSV file (spreadsheet) that can be passed in to `sourmash plot` with `--labels-from` in order to customize the labels. **Note:** compare by default produces a symmetric similarity matrix that can be used for clustering in downstream tasks. With `--containment`, however, this matrix is no longer symmetric and cannot formally be used for clustering (although `sourmash plot` will still cluster it). The containment matrix is organized such that the value in row A for column B is the containment of the B'th sketch in the A'th sketch, i.e. ``` C(A, B) = B.contained_by(A) ``` **Note:** The ANI estimate will be calculated based on Jaccard similarity by default; however, if `--containment`, `--max-containment`, or `--avg-containment` is specified, those values will be used instead. With `--containment --ani`, the ANI output matrix will be asymmetric as discussed above. ### `sourmash plot` - cluster and visualize comparisons of many signatures **Note:** The [betterplot plugin](https://github.com/sourmash-bio/sourmash_plugin_betterplot/) provides a wide array of improved visualization options, including MDS and tSNE plots, as well as more customizable version of `plot`. Please take a look! The `plot` subcommand produces two plots -- a dendrogram and a dendrogram+matrix -- from a matrix created by `sourmash compare --output `. The default output is two PNG files. Usage: ``` sourmash plot ``` Options: * `--pdf` -- output PDF files. (defaults to PNG) * `--labels` -- display the signature names on the plot (default) * `--indices` -- turn on index display on the plot. * `--vmax` -- maximum value (default 1.0) for heatmap. * `--vmin` -- minimum value (default 0.0) for heatmap. * `--subsample=` -- plot a maximum of samples, randomly chosen. * `--subsample-seed=` -- seed for pseudorandom number generator. Example command lines for label and index display - * `--indices` will show only numbers; * `--no-labels --no-indices` will remove all labels! Example output: ![An E. coli comparison plot](_static/ecoli_cmp.matrix.png) ### `sourmash search` - search for signatures in collections or databases The `search` subcommand searches a collection of signatures (in any of the [formats supported by sourmash](#storing-and-searching-signatures)) for matches to a single query signature. It can search for matches with either high [Jaccard similarity](https://en.wikipedia.org/wiki/Jaccard_index) or containment; the default is to use Jaccard similarity, unless `--containment` is specified. `-o/--output` will create a CSV file containing all of the matches with respective similarity or containment score. `search` makes use of [indexed databases](#loading-many-signatures) to decrease search time and memory where possible. Usage: ``` sourmash search query.sig [ ... ] ``` Example output: ``` % sourmash search tests/test-data/47.fa.sig gtdb-rs207.genomic-reps.dna.k31.zip ... -- loaded 65703 total signatures from 1 locations. after selecting signatures compatible with search, 65703 remain. 2 matches above threshold 0.080: similarity match ---------- ----- 32.3% GCF_900456975.1 Shewanella baltica strain=NCTC10735, 5088... 14.0% GCF_002838165.1 Shewanella sp. Pdp11 strain=Pdp11, ASM283... ``` `search` takes a number of command line options - * `--containment` - find matches using the containment index rather than Jaccard similarity; * `--max-containment` - find matches using the max containment index rather than Jaccard similarity; * `-t/--threshold` - lower threshold for matching; defaults to 0.08; * `--best-only` - find and report only the best match; * `-n/--num-results` - number of matches to report to stdout; defaults to 3; 0 to report all; Match information can be saved to a CSV file with `-o/--output`; with `-o`, all matches above the threshold will be saved, not just those printed to stdout (which are limited to `-n/--num-results`). The `--containment` flag calculates the containment of the query in database matches; this is an asymmetric order-dependent measure, unlike Jaccard. Here, `search --containment Q A B C D` will report the containment of `Q` in each of `A`, `B`, `C`, and `D`. This is opposite to the order used by `prefetch`, where the composite sketch (e.g. metagenomes) is the query, and the matches are contained items (e.g. genomes). As of sourmash 4.2.0, `search` supports `--picklist`, to [select a subset of signatures to search, based on a CSV file](#using-picklists-to-subset-large-collections-of-signatures). This can be used to search only a small subset of a large collection, or to exclude a few signatures from a collection, without modifying the collection itself. Related commands: * `sourmash compare` will compare many sketches to many sketches. * `sourmash prefetch` is an upgraded version of `search` that returns more information in the output CSV file. * `sourmash scripts manysearch` from [the branchwater plugin](https://github.com/sourmash-bio/sourmash_plugin_branchwater) will search multiple sketches against a database, and outputs abundance-weighted comparisons as well. ### `sourmash gather` - find metagenome members **Note:** As of 2025, we have a much faster implementation of `gather` called `fastgather` available in [the branchwater plugin](https://github.com/sourmash-bio/sourmash_plugin_branchwater). It is multithreaded and similar in memory usage, although it does accept a slightly more restricted set of inputs than `gather`. The `gather` subcommand selects the best reference genomes to use for a metagenome analysis, by finding the smallest set of non-overlapping matches to the query metagenome in a database of genomes. This is specifically meant for metagenome and genome bin analysis. (See [Classifying Signatures](classifying-signatures.md) for more information on the different approaches that can be used here.) `sourmash gather` takes exactly one query and one or more [collections of signatures](#storing-and-searching-signatures). Please see [`sourmash multigather`](#sourmash-multigather-do-gather-with-many-queries) if you have multiple queries! If the input signature was created with `-p abund`, output will be abundance weighted (unless `--ignore-abundances` is specified). `-o/--output` will create a CSV file containing the matches and quite a bit more information; see [the CSV output file documentation for gather](classifying-signatures.md#appendix-d-gather-csv-output-columns) for details of the output columns. `gather`, like `search`, works with any of the [signature collection formats supported by sourmash](#storing-and-searching-signatures) and will make use of [indexed databases](#loading-many-signatures) to decrease search time and memory where possible. We recommend using RocksDB indexes if you are running gather regularly! Usage: ``` sourmash gather query.sig [ ... ] ``` Example output: ``` overlap p_query p_match --------- ------- -------- 1.4 Mbp 11.0% 58.0% JANA01000001.1 Fusobacterium sp. OBRC... 1.0 Mbp 7.7% 25.9% CP001957.1 Haloferax volcanii DS2 pla... 0.9 Mbp 7.4% 11.8% BA000019.2 Nostoc sp. PCC 7120 DNA, c... 0.7 Mbp 5.9% 23.0% FOVK01000036.1 Proteiniclasticum rumi... 0.7 Mbp 5.3% 17.6% AE017285.1 Desulfovibrio vulgaris sub... ... found less than 50.0 kbp in common. => exiting found 64 matches total; the recovered matches hit 94.0% of the abundance-weighted query. the recovered matches hit 45.6% of the query k-mers (unweighted). ``` For each match, * 'overlap', the first column, is the estimated number of base pairs shared between the match and the query, based on the number of shared hashes. * 'p_query' is the _percentage_ of the query that overlaps with the match; it is the amount of the metagenome "explained" by this match. It is typically a lower bound on the percent of metagenomes reads that will map to this genome. * 'p_match' is the percentage of the _match_ that overlaps with the query; it is the "detection" of the match in the metagenome. It is typically a lower bound on the number of base pairs that will be covered by read mapping. Quite a bit more information per match row is available in the CSV output saved with `-o`; for details, see [Classifying signatures: how sourmash gather works](classifying-signatures.md#appendix-a-how-sourmash-gather-works). The "recovered matches" lines detail how much of the query is explained by the entire collection of matches. You will get two numbers if your metagenome sketch has been calculated with `-p abund`, and only one if it does not have abundances. The abundance-weighted number should approximate the fraction of metagenome reads that will map to at least one reference genome, while the unweighted number describes how much of the metagenome itself matches to genomes. Here's another way to put it: if the metagenome could be perfectly assembled into contigs, the unweighted number would approximate the number of bases from the contigs that would match perfectly to at least one genome in the reference database. More practically, the abundance-weighted number is less sensitive to sequencing errors. See [classifying signatures](classifying-signatures.md#abundance-weighting) or [the FAQ](faq.md) for more information! The command line option `--threshold-bp` sets the threshold below which matches are no longer reported; by default, this is set to 50kb. See the Appendix in [Classifying Signatures](classifying-signatures.md) for details. As of sourmash 4.2.0, `gather` supports `--picklist`, to [select a subset of signatures based on a CSV file](#using-picklists-to-subset-large-collections-of-signatures). This can be used to search only a small subset of a large collection, or to exclude a few signatures from a collection, without modifying the collection itself. Note: Use `sourmash gather` to analyze a metagenome against a collection of genomes. Then use `sourmash tax metagenome` to integrate that collection of genomes with taxonomic information. #### Alternative search mode for low-memory (but slow) search: `--linear` By default, `sourmash gather` uses all information available for faster search. In particular, for SBTs, `prefetch` will prune the search tree. This can be slow and/or memory intensive for very large databases, and `--linear` asks `sourmash prefetch` to instead use a linear search across all leaf nodes in the tree. The results are the same whether `--no-linear` or `--linear` is used. #### Alternative search mode: `--no-prefetch` By default, `sourmash gather` does a "prefetch" to find *all* candidate signatures across all databases, before removing overlaps between the candidates. In rare circumstances, depending on the databases and parameters used, this may be slower or more memory intensive than doing iterative overlap removal. Prefetch behavior can be turned off with `--no-prefetch`. The results are the same whether `--prefetch` or `--no-prefetch` is used. This option can be used with or without `--linear` (although `--no-prefetch --linear` will generally be MUCH slower). ### `sourmash index` - build an index of signatures The `sourmash index` command creates indexed databases from a collection of signatures. This can be used to create databases from private collections of genomes or metagenomes, and can also be used to create databases for e.g. subsets of GenBank. As of sourmash v4.9.0, there are two types of indexed databases supported by sourmash: RocksDB-based inverted indexes, and Sequence Bloom Trees (SBTs). RocksDB indexes are a newer form of fast and low-memory on-disk indexes that work at massive scale, while SBTs are older indexes that we support for legacy reasons. All signatures in an index must be of compatible types (i.e. the same k-mer size, scaled, and molecule type). You can specify the usual command line selectors (`-k`, `--scaled`, `--dna/--protein/--hp/--dayhoff/--skipm1n3/--skipm2n3`, etc.) to pick out the types of signatures to include when running `index`. Usage: ``` sourmash index .rocksdb [ ... ] -F rocksdb ``` This will create a `database.rocksdb` file containing the sketches. Note that you can use `--from-file` to pass `index` a text file containing a list of file names to index; you can also provide individual signature files, directories full of signatures, or other sourmash databases. As of sourmash 4.2.0, `index` supports `--picklist`, to [select a subset of signatures based on a CSV file](#using-picklists-to-subset-large-collections-of-signatures). This can be used to index a subset of a large collection, or to exclude a few signatures from an index being built from a large collection. Note: until sourmash v5, the default index type is "SBT"; this is to ensure backwards compatibility of the command line API for semantic versioning. We no longer recommend SBTs. Also note that a third index type, 'zip', is supported by the command line, but this should be used solely for testing purposes - it is identical in format to a zipped database output by e.g. `sig cat`, but restricted to the same ksize/moltype/scaled. ### `sourmash prefetch` - select subsets of very large databases for more processing The `prefetch` subcommand searches a scaled signature (usually a genome or a metagenome) for matches in a large database of genomes, using containment. It is similar to `search --containment`, while taking a `--threshold-bp` argument like `gather` does for thresholding matches (instead of using Jaccard similarity or containment). Note that `prefetch` uses the composite sketch (e.g. a metagenome) as the query, and finds all matching subjects (e.g. genomes) from the database - the arguments are in the opposite order from `search --containment`. Note that the `manysearch` command in [the branchwater plugin](https://github.com/sourmash-bio/sourmash_plugin_branchwater) is a fast, multithreaded version of `prefetch` that will search for multiple genomes in multiple metagenomes. `sourmash prefetch` is intended to select a subset of a large database for further processing. As such, it can search very large collections of signatures (potentially millions or more), operates in very low memory (see `--linear` option, below), and does no post-processing of signatures. `prefetch` has four main output options, which can all be used individually or together: * `-o/--output` produces a CSV summary file; see [the CSV output file documentation for prefetch](classifying-signatures.md#appendix-e-prefetch-csv-output-columns) for details of the output columns. * `--save-matches` saves all matching signatures; * `-save-matching-hashes` saves a single signature containing all of the hashes that matched any signature in the database at or above the specified threshold; * `--save-unmatched-hashes` saves a single signature containing the complement of `--save-matching-hashes`. Other options include: * the usual `-k/--ksize` and `--dna/--protein/--dayhoff/--hp/--skipm1n3/--skipm2n3` signature selectors; * `--threshold-bp` to require a minimum estimated bp overlap for output; * `--scaled` for downsampling; * `--force` to continue past survivable errors; * `--picklist` will select a subset of signatures to search, using [a picklist](#using-picklists-to-subset-large-collections-of-signatures) #### Alternative search mode for low-memory (but slow) search: `--linear` By default, `sourmash prefetch` uses all information available for faster search. In particular, for SBTs, `prefetch` will prune the search tree. This can be slow and/or memory intensive for very large databases, and `--linear` asks `sourmash prefetch` to instead use a linear search across all leaf nodes in the tree. #### Caveats and comments `sourmash prefetch` provides no guarantees on output order. It runs in "streaming mode" on its inputs, in that each input file is loaded, searched, and then unloaded. And `sourmash prefetch` can be run separately on multiple databases, after which the results can be searched in combination with `search`, `gather`, `compare`, etc. A motivating use case for `sourmash prefetch` is to run it on multiple large databases with a metagenome query using `--threshold-bp=0`, `--save-matching-hashes matching-hashes.sig`, and `--save-matches db-matches.sig`, and then run `sourmash gather matching-hashes.sig db-matches.sig`. This combination of commands ensures that the more time- and memory-intensive `gather` step is run only on a small set of relevant signatures, rather than all the signatures in the database. ### `sourmash multigather` - do gather with many queries **Note:** As of 2025, we have a much faster implementation of `multigather` called `fastmultigather` available in [the branchwater plugin](https://github.com/sourmash-bio/sourmash_plugin_branchwater). It is multithreaded and similar in memory usage, although it does accept a slightly more restricted set of inputs than `multigather`. The `multigather` subcommand runs `sourmash gather` on multiple queries. (See [`sourmash gather` docs](#sourmash-gather-find-metagenome-members) for specifics on what gather does, and how!) Usage: ``` sourmash multigather --query --db ``` Note that multigather is single threaded, so it offers no substantial efficiency gains over just running gather multiple times! Nonetheless, it is useful for situations where you have many sketches organized in a combined file, e.g. sketches built with `sourmash sketch ... --singleton`). #### `multigather` output files multigather produces three output files for each query: * `.csv` - gather CSV output * `.matches.sig` - all matching outputs * `.unassigned.sig` - all remaining unassigned hashes As of sourmash v4.8.7, `` is set as follows: * the filename attribute of the query sketch, if it is not empty or `-`; * the query sketch md5sum, if the query filename is empty or `-`; * the query filename + the query sketch md5sum (`.`), if `-U/--output-add-query-md5sum` is specified; By default, `multigather` will complain and exit with an error if the same `` is used repeatedly and an output file is going to be overwritten. With `-U/--output-add-query-md5sum` this should only happen when identical sketches are present in a query database. Use `--force-allow-overwrite-output` to allow overwriting of output files without an error. ## `sourmash tax` subcommands for integrating taxonomic information into gather results The `sourmash tax` subcommands support taxonomic analysis of genomes and taxonomic profiling of metagenomes. See [taxonomic profiling with sourmash](classifying-signatures.md#taxonomic-profiling-with-sourmash) for more information. There are a number of plotting and visualization options for `tax metagenome` output available in [the betterplot plugin](#the-betterplot-plugin-improved-plotting-and-visualization) as well as [the taxburst fork of the Krona visualization software](https://taxburst.github.io/). The sourmash `tax` or `taxonomy` commands integrate taxonomic information with the results of `sourmash gather`. All `tax` commands require one or more properly formatted `taxonomy` files where the identifiers correspond to those in the database(s) used for `gather`. Note that if using multiple databases, the `gather` needs to have been conducted against all desired databases within the same `gather` command (we cannot combine separate `gather` runs for the same query). For supported databases (e.g. GTDB, NCBI), we provide taxonomy csv files, but they can also be generated for user-generated databases. As of v4.8 and 4.8.6, respectively, some sourmash taxonomy commands can also use `LIN` or `ICTV` lineage information. `tax` commands rely upon the fact that `gather` provides both the total fraction of the query matched to each database matched, as well as a non-overlapping `f_unique_to_query`, which is the fraction of the query uniquely matched to each reference genome. The `f_unique_to_query` for any reference match will always be between (0% of query matched) and 1 (100% of query matched), and for a query matched to multiple references, the `f_unique_to_query` will sum to at most 1 (100% of query matched). We use this property to aggregate gather matches at the desired taxonomic rank. For example, if the gather results for a metagenome include results for 30 different strains of a given species, we can sum the fraction uniquely matched to each strain to obtain the fraction uniquely matched to this species. Alternatively, taxonomic summarization can take into account abundance weighting; see [classifying signatures](classifying-signatures.md) for more information. As with all reference-based analysis, results can be affected by the completeness of the reference database. However, summarizing taxonomic results from `gather` minimizes issues associated with increasing size and redundancy of reference databases. For more details on how `gather` works and can be used to classify signatures, see ### `sourmash tax metagenome` - summarize metagenome content from `gather` results `sourmash tax metagenome` summarizes gather results for each query metagenome by taxonomic lineage. Here is an example command to summarize a single `gather csv`, where the query was gathered against `gtdb-rs202` representative species database: ``` sourmash tax metagenome --gather-csv HSMA33MX_gather_x_gtdbrs202_k31.csv \ --taxonomy gtdb-rs202.taxonomy.v2.csv ``` One or more output formats can be provided with `-F/--output-format`, and output will be saved to files named for each format. The possible output formats are listed below, followed by the file extension used when writing to a file rather than stdout. When using more than one output format, you must provide an output basename (`--output-base`) that will be used to name the output files. If an `--output-dir` is provided, files will output to that directory. - `human`: ".human.txt" - `csv_summary`: ".summarized.csv" - `lineage_summary`: ".lineage_summary.tsv" - `krona`: ".krona.tsv" - `kreport`: ".kreport.txt" - `lingroup`: ".lingroup.tsv" - `bioboxes`: ".bioboxes.profile" In general, `tax metagenome` should be used with gather results that use abundance (e.g. sketches calculated with `-p abund`). This will properly report the proportion of the metagenome data that can be classified. However, in sourmash v4, abundance reporting is inconsistent between formats; see the below table for a summary. As of sourmash v4.9.4, we recommend using `--use-abund` with `tax metagenome`, which will force consistent reporting of abundances, and will fail when abundances are not available. `--ignore-abund` can be used to force ignoring abundances. `--use-abund` will be the default behavior in sourmash v5. | output format | v4 behavior | v5 default behavior | notes | | -------- | -------- | -------- | -- | | human | abund | abund | default output in v5 | csv_summary | provides both | provides both | default output in v4 | lineage_summary | no abund | abund | | krona | no abund | abund | | kreport | abund | abund | :::{warning} **sourmash v5 breaking change:** In sourmash v5.0 and later, the default output format will become `human`, instead of `csv_summary`. Likewise, `--use-abundances` will be on by default for `tax metagenome`, and it will be an error to not have abundances in your gather output. You can specify `--ignore-abundances` to override the error. ::: You can specify `--v4` to keep the old behavior, or `--v5` to support the new default behavior, or simply specify the desired formats and/or abundance flags to override the default. See [Migrating from sourmash v4.x to sourmash v5.x](support.md#migrating-from-sourmash-v4x-to-sourmash-v5x) for more details. #### `human` output format `human` will be the default output format in sourmash v5. This outputs a human-readable table with summarization for each leaf node. It is not intended for automated parsing. example `human` output: ``` sample name proportion cANI lineage ----------- ---------- ---- ------- test1 86.9% - unclassified test1 5.8% 92.5% d__Bacteria;p__Proteobacteria;c__Gammaproteobacteria;o__Enterobacterales;f__Enterobacteriaceae;g__Escherichia;s__Escherichia coli test1 5.7% 92.5% d__Bacteria;p__Bacteroidota;c__Bacteroidia;o__Bacteroidales;f__Bacteroidaceae;g__Prevotella;s__Prevotella copri test1 1.6% 89.1% d__Bacteria;p__Bacteroidota;c__Bacteroidia;o__Bacteroidales;f__Bacteroidaceae;g__Phocaeicola;s__Phocaeicola vulgatus ``` #### `csv_summary` output format `csv_summary` is the default output format in sourmash v4. This outputs a `csv` with lineage summarization for each taxonomic rank. This output currently consists of six columns, `query_name,rank,fraction,lineage,query_md5,query_filename`, where `fraction` is the fraction of the query matched to the reported rank and lineage. example `csv_summary` output from the command above: ``` query_name,rank,fraction,lineage HSMA33MX,superkingdom,0.131,d__Bacteria HSMA33MX,phylum,0.073,d__Bacteria;p__Bacteroidota HSMA33MX,phylum,0.058,d__Bacteria;p__Proteobacteria . . . HSMA33MX,species,0.058,d__Bacteria;p__Proteobacteria;c__Gammaproteobacteria; o__Enterobacterales;f__Enterobacteriaceae;g__Escherichia;s__Escherichia coli HSMA33MX,species,0.057,d__Bacteria;p__Bacteroidota;c__Bacteroidia; o__Bacteroidales;f__Bacteroidaceae;g__Prevotella;s__Prevotella copri HSMA33MX,species,0.016,d__Bacteria;p__Bacteroidota;c__Bacteroidia; o__Bacteroidales;f__Bacteroidaceae;g__Phocaeicola;s__Phocaeicola vulgatus ``` The `query_md5` and `query_filename` columns are omitted here for brevity. Note: When using `--lins` with a `--lingroup` file, the `csv_summary` file will report summarization for each specified `lingroup`, rather than all possible `lin` ranks (v4.8.12+). #### `krona` output format `krona` format is a tab-separated list of these results at a specific rank. The first column, `fraction` is the fraction of the query matched to the reported rank and lineage. The remaining columns are `superkingdom`, `phylum`, ... etc down to the rank used for summarization. This output can be used directly for summary visualization. To generate `krona`, we add `--output-format krona` to the command above, and need to specify a rank to summarize. Here's the command for reporting `krona` summary at `species` level: ``` sourmash tax metagenome --gather-csv HSMA33MX_gather_x_gtdbrs202_k31.csv \ --taxonomy gtdb-rs202.taxonomy.v2.csv \ --output-format krona --rank species ``` example krona output from this command: ``` fraction superkingdom phylum class order family genus species 0.05815279361459521 Bacteria Proteobacteria Gammaproteobacteria Enterobacterales Enterobacteriaceae Escherichia Escherichia coli 0.05701254275940707 Bacteria Bacteroidetes Bacteroidia Bacteroidales Prevotellaceae Prevotella Prevotella copri 0.015637726014008795 Bacteria Bacteroidetes Bacteroidia Bacteroidales Bacteroidaceae Bacteroides Bacteroides vulgatus ``` #### `lineage_summary` output format The lineage summary format is most useful when comparing across metagenome queries. Each row is a lineage at the desired reporting rank. The columns are each query used for gather, with the fraction match reported for each lineage. This format is commonly used as input for many external multi-sample visualization tools. To generate `lineage_summary`, we add `--output-format lineage_summary` to the summarize command, and need to specify a rank to summarize. Here's the command for reporting `lineage_summary` for two queries (HSMA33MX, PSM6XBW3) summary at `species` level. ``` sourmash tax metagenome --gather-csv HSMA33MX_gather_x_gtdbrs202_k31.csv \ --gather-csv PSM6XBW3_gather_x_gtdbrs202_k31.csv \ --taxonomy gtdb-rs202.taxonomy.v2.csv \ --output-format lineage_summary --rank species ``` example `lineage_summary`: ``` lineage HSMA33MX PSM6XBW3 d__Bacteria;p__Bacteroidota;c__Bacteroidia;o__Bacteroidales;f__Bacteroidaceae;g__Phocaeicola;s__Phocaeicola vulgatus 0.015637726014008795 0.015642822225843248 d__Bacteria;p__Bacteroidota;c__Bacteroidia;o__Bacteroidales;f__Bacteroidaceae;g__Prevotella;s__Prevotella copri 0.05701254275940707 0.05703112269838684 d__Bacteria;p__Proteobacteria;c__Gammaproteobacteria;o__Enterobacterales;f__Enterobacteriaceae;g__Escherichia;s__Escherichia coli 0.05815279361459521 0.05817174515235457 ``` To produce multiple output types from the same command, add the types into the `--output-format` argument, e.g. `--output-format summary krona lineage_summary` #### `kreport` output format The `kreport` output reports kraken-style `kreport` output, which may be useful for comparison with other taxonomic profiling methods. While this format typically records the percent of number of reads assigned to taxa, we create ~comparable output by reporting the percent of k-mers matched to each taxon and the estimated number of base pairs that these k-mers represent. To best represent the percent of all reads, we use k-mer abundance information in this output. To generate this properly, query FracMinHash sketches should be generated with abundance information (`-p abund`) to allow abundance-weighted `gather` results. Note: `sourmash gather` makes all assignments to genomes, and then `sourmash tax` integrates taxonomy information and uses LCA-style summarization to build assignments. For species-level specificity, our current recommendation is to use use our default k-mer size of 31. standard `kreport` columns (read-based tools): - `Percent Reads Contained in Taxon`: The cumulative percentage of reads for this taxon and all descendants. - `Number of Reads Contained in Taxon`: The cumulative number of reads for this taxon and all descendants. - `Number of Reads Assigned to Taxon`: The number of reads assigned directly to this taxon (not a cumulative count of all descendants). - `Rank Code`: (U)nclassified, (R)oot, (D)omain, (K)ingdom, (P)hylum, (C)lass, (O)rder, (F)amily, (G)enus, or (S)pecies. - `NCBI Taxon ID`: Numerical ID from the NCBI taxonomy database. - `Scientific Name`: The scientific name of the taxon. Example reads-based `kreport` with all columns: ``` 88.41 2138742 193618 K 2 Bacteria 0.16 3852 818 P 201174 Actinobacteria 0.13 3034 0 C 1760 Actinomycetia 0.13 3034 45 O 85009 Propionibacteriales 0.12 2989 1847 F 31957 Propionibacteriaceae 0.05 1142 352 G 1912216 Cutibacterium 0.03 790 790 S 1747 Cutibacterium acnes ``` sourmash `kreport` columns: - `Percent [k-mers] contained in taxon`: The cumulative percentage of k-mers for this taxon and all descendants. - `Estimated base pairs contained in taxon`: The cumulative estimated base pairs for this taxon and all descendants. - `Estimated base pairs "assigned" (species-level)`: The estimated base pairs assigned at species-level (cumulative count of base pairs assigned to individual genomes in this species). - `Rank Code`: (U)nclassified, (R)oot, (D)omain, (K)ingdom, (P)hylum, (C)lass, (O)rder, (F)amily, (G)enus, or (S)pecies. - `NCBI Taxon ID`: Reported (v4.7+) if using NCBI taxonomy. Otherwise blank. - `Scientific Name`: The scientific name of the taxon. notes: - `gather` assigns k-mers to specific genomes. To mimic the output of other tools, we report all results as "assigned" to species-level, which summarizes the k-mers matched to each genome within a given species. Hence, column 3 will show all estimated base pairs at this level, and 0 for all other ranks. Column 2 contains the summarized info at the higher ranks. - Since `gather` results are non-overlapping and all assignments are done at the genome level, the percent match (first column) will sum to 100% at each rank (aside from rounding issues) when including the unclassified (U) percentage. Higher-rank assignments are generated using LCA-style summarization of genome matches. - Rows are ordered by rank and then ~percent containment. example sourmash `{output-name}.kreport.txt`: ``` 92.73 64060000 D Bacteria 0.44 11299000 D Eukaryota 6.82 284315000 U unclassified 60.23 30398000 P Proteobacteria 21.86 22526000 P Firmicutes 10.41 5250000 P Bacteroidetes . . . 3.94 6710000 S Escherichia coli 4.56 6150000 S Pseudomonas aeruginosa 0.71 5801000 S Clostridium beijerinckii 2.55 5474000 S Bacillus cereus 21.95 4987000 S Escherichia sp. XD7 28.57 4124000 S Cereibacter sphaeroides 0.25 4014000 S Acinetobacter baumannii 7.23 3934000 S Staphylococcus haemolyticus 0.09 3187000 S Phocaeicola vulgatus 0.61 2820000 S Streptococcus agalactiae 0.20 2499000 S Cutibacterium acnes 0.03 2339000 S Deinococcus radiodurans 10.31 2063000 S Porphyromonas gingivalis 9.24 2011000 S Streptococcus mutans ``` #### `lingroup` output format When using LIN taxonomic information, you can optionally also provide a `lingroup` file with two required columns: `name` and `lin`. If provided, we will produce a file, `{base}.lingroups.tsv`, where `{base}` is the name provided via the `-o`,` --output-base` option. This output will select information from the full summary that match the LIN prefixes provided as groups. This output format consists of four columns: - `name`, `lin` columns are taken directly from the `--lingroup` file - `percent_containment`, the total percent of the dataset contained in this lingroup and all descendants - `num_bp_contained`, the estimated number of base pairs contained in this lingroup and all descendants. Similar to `kreport` above, we use the wording "contained" rather than "assigned," because `sourmash` assigns matches at the genome level, and the `tax` functions summarize this information. example output: ``` name lin percent_containment num_bp_contained lg1 0;0;0 5.82 714000 lg2 1;0;0 5.05 620000 lg3 2;0;0 1.56 192000 lg3 1;0;1 0.65 80000 lg4 1;0;1;0;0;0;0;0;0;0;0;0;0;0;0;0;0;0;0;0 0.65 80000 ``` Related lingroup subpaths will be grouped in output, but exact ordering may change between runs. Note: this output format requires a single sample only. For a similar output with multiple query samples, provide the `lingroup` file and use the 'csv_summary' output format. #### `bioboxes` output format When using standard taxonomic ranks (not lins), you can choose to output a 'bioboxes' profile, `{base}.bioboxes.profile`, where `{base}` is the name provided via the `-o/--output-base` option. This output is organized according to the [bioboxes profile specifications](https://github.com/bioboxes/rfc/tree/master/data-format) so that this file can be used for CAMI challenges. As of v4.9.4, `bioboxes` will support taxonomies with missing ranks, as seen in some NCBI taxonomies (e.g. taxpath: 2|1239|||||2292892|). In these cases, the missing rank-taxid combinations will be omitted from the output. This output format starts with some header information: ``` #CAMI Submission for Taxonomic Profiling @Version:0.9.3 @SampleID:SAMPLEID @Ranks:superkingdom|phylum|class|order|family|genus|species|strain @__program__:sourmash @@TAXID RANK TAXPATH TAXPATHSN PERCENTAGE ``` and then provides taxonomic profiling information in the tab-separated columns described by the last header line: - `TAXID` - specifies a unique alphanumeric ID for a node in a reference tree such as the NCBI taxonomy - `RANK` - superkingdom --> strain - `TAXPATH` - the path from the root of the reference taxonomy to the respective taxon - `TAXPATHSN` - scientific names of taxpath - `PERCENTAGE` (0-100) - field specifies what percentage of the sample was assigned to the respective TAXID example output (using small test data): ``` # Taxonomic Profiling Output @SampleID:test1 @Version:0.10.0 @Ranks:superkingdom|phylum|class|order|family|genus|species @__program__:sourmash @@TAXID RANK TAXPATH TAXPATHSN PERCENTAGE 2 superkingdom 2 Bacteria 13.08 976 phylum 2|976 Bacteria|Bacteroidota 7.27 1224 phylum 2|1224 Bacteria|Pseudomonadota 5.82 200643 class 2|976|200643 Bacteria|Bacteroidota|Bacteroidia 7.27 1236 class 2|1224|1236 Bacteria|Pseudomonadota|Gammaproteobacteria 5.82 171549 order 2|976|200643|171549 Bacteria|Bacteroidota|Bacteroidia|Bacteroidales 7.27 91347 order 2|1224|1236|91347 Bacteria|Pseudomonadota|Gammaproteobacteria|Enterobacterales 5.82 171552 family 2|976|200643|171549|171552 Bacteria|Bacteroidota|Bacteroidia|Bacteroidales|Prevotellaceae 5.70 543 family 2|1224|1236|91347|543 Bacteria|Pseudomonadota|Gammaproteobacteria|Enterobacterales|Enterobacteriaceae 5.82 815 family 2|976|200643|171549|815 Bacteria|Bacteroidota|Bacteroidia|Bacteroidales|Bacteroidaceae 1.56 838 genus 2|976|200643|171549|171552|838 Bacteria|Bacteroidota|Bacteroidia|Bacteroidales|Prevotellaceae|Prevotella 5.70 561 genus 2|1224|1236|91347|543|561 Bacteria|Pseudomonadota|Gammaproteobacteria|Enterobacterales|Enterobacteriaceae|Escherichia 5.82 909656 genus 2|976|200643|171549|815|909656 Bacteria|Bacteroidota|Bacteroidia|Bacteroidales|Bacteroidaceae|Phocaeicola 1.56 165179 species 2|976|200643|171549|171552|838|165179 Bacteria|Bacteroidota|Bacteroidia|Bacteroidales|Prevotellaceae|Prevotella|Prevotella copri 5.70 562 species 2|1224|1236|91347|543|561|562 Bacteria|Pseudomonadota|Gammaproteobacteria|Enterobacterales|Enterobacteriaceae|Escherichia|Escherichia coli 5.82 821 species 2|976|200643|171549|815|909656|821 Bacteria|Bacteroidota|Bacteroidia|Bacteroidales|Bacteroidaceae|Phocaeicola|Phocaeicola vulgatus 1.56 ``` ### `sourmash tax genome` - classify a genome using `gather` results `sourmash tax genome` reports likely classification for each query, based on `gather` matches. By default, classification requires at least 10% of the query to be matched. Thus, if 10% of the query was matched to a species, the species-level classification can be reported. However, if 7% of the query was matched to one species, and an additional 5% matched to a different species in the same genus, the genus-level classification will be reported. `sourmash tax genome` can use an ANI threshold (`--ani-threshold`) instead of a containment threshold. This works the same way as the containment threshold (and indeed, is using the same underlying information). Note that for DNA k-mers, k=21 ANI is most similar to alignment-based ANI values, and ANI values should only be compared if they were generated using the same ksize. Optionally, `genome` can instead report classifications at a desired `rank`, regardless of match threshold (`--rank` argument, e.g. `--rank species`). If using `--lins` taxonomy, you can also provide a `--lingroup` file containing two columns, `name`, and `lin`, which provide a series of lin prefixes of interest. If provided, genome classification will be restricted to provided lingroups only. All other options (`--rank`, `--ani-threshold`, etc) should continue to function. If you specify a `--rank` that does not have an associated lingroup, sourmash will notify you that you eliminated all classification options. Note that these thresholds and strategies are under active testing. To illustrate the utility of `genome`, let's consider a signature consisting of two different Shewanella strains, `Shewanella baltica OS185 strain=OS185` and `Shewanella baltica OS223 strain=OS223`. For simplicity, we gave this query the name "Sb47+63". When we gather this signature against the `gtdb-rs202` representatives database, we see 66% matches to one strain, and 33% to the other: abbreviated gather_csv: ``` f_match,f_unique_to_query,name,query_name 0.664,0.664,"GCF_000021665.1 Shewanella baltica OS223 strain=OS223, ASM2166v1",Sb47+63 0.656,0.335,"GCF_000017325.1 Shewanella baltica OS185 strain=OS185, ASM1732v1",Sb47+63 ``` > Here, `f_match` shows that independently, both strains match ~65% percent of this mixed query. The `f_unique_to_query` column has the results of gather-style decomposition. As the OS223 strain had a slightly higher `f_match` (66%), it was the first match. The remaining 33% of the query matched to strain OS185. We can use `tax genome` on this gather csv to classify our "Sb47+63" mixed-strain query: ``` sourmash tax genome --gather-csv 47+63_x_gtdb-rs202.gather.csv \ --taxonomy gtdb-rs202.taxonomy.v2.csv ``` > This command uses the default classification strategy, which uses a containment threshold of 0.1 (10%). `sourmash tax genome` can produce the following output formats: - `human`: ".human.txt", - `csv_summary`: ".classifications.csv", - `krona`: ".krona.tsv", - `lineage_summary`: ".lineage_summary.tsv", :::{warning} **sourmash v5 breaking change:** In sourmash v5.0 and later, the default output format will become `human`, instead of `csv_summary`. You can specify `--v4` to keep the old behavior, or `--v5` to support the new default behavior, or simply specify the desired output format. See [Migrating from sourmash v4.x to sourmash v5.x](support.md#migrating-from-sourmash-v4x-to-sourmash-v5x) for more details. ::: #### `csv_summary` output format `csv_summary` is the default output format. This outputs a `csv` with taxonomic classification for each query genome. This output currently consists of six columns, `query_name,rank,fraction,lineage,query_md5,query_filename`, where `fraction` is the fraction of the query matched to the reported rank and lineage. The `status` column provides additional information on the classification: - `match` - this query was classified - `nomatch`- this query could not be classified - `below_threshold` - this query was classified at the specified rank, but the query fraction matched was below the containment threshold Here is the `csv_summary` output from classifying this mixed-strain Shewanella query to species level: ``` query_name,status,rank,fraction,lineage "Sb47+63",match,species,1.000,d__Bacteria;p__Proteobacteria;c__Gammaproteobacteria;o__Enterobacterales;f__Shewanellaceae;g__Shewanella;s__Shewanella baltica ``` >Here, we see that the match percentages to both strains have been aggregated, and we have 100% species-level `Shewanella baltica` annotation. We have omitted the `query_md5` and `query_filename` columns for brevity. #### `krona` output format `krona` format is a tab-separated list of these results at a specific rank. The first column, `fraction` is the fraction of the query matched to the reported rank and lineage. The remaining columns are `superkingdom`, `phylum`, ... etc down to the rank used for summarization. This output can be used directly for `krona` visualization. To generate `krona`, we must classify by `--rank` instead of using the classification threshold. For the command, we add `--output-format krona` and `--rank ` to the command above. Here's the command for producing `krona` output for `species`-level classifications: ``` sourmash tax genome --gather-csv Sb47+63_gather_x_gtdbrs202_k31.csv \ --taxonomy gtdb-rs202.taxonomy.v2.csv \ --output-format krona --rank species ``` > Note that specifying `--rank` forces classification by rank rather than by the containment threshold. Here is the `krona`-formatted output for this command: ``` fraction superkingdom phylum class order family genus species 1.0 d__Bacteria p__Proteobacteria c__Gammaproteobacteria o__Enterobacterales f__Shewanellaceae g__Shewanella s__Shewanella baltica ``` To produce multiple output types from the same command, add the types into the `--output-format` argument, e.g. `--output-format csv_summary krona`. **Note that specifying the classification rank with `--rank`, (e.g. `--rank species`), as needed for `krona` output, forces classification by `rank` rather than by containment threshold.** If the query classification at this rank does not meet the containment threshold (default=0.1), the `status` column will contain `below_threshold`. ### `sourmash tax annotate` - annotates gather output with taxonomy `sourmash tax annotate` adds a column with taxonomic lineage information for each genome match in the gather output, without LCA summarization or classification. This format is not required for either `metagenome` or `genome`, but may be helpful for other downstream analyses. By default, `annotate` uses the name of each input gather csv to write an updated version with lineages information. For example, annotating `sample1.gather.csv` would produce `sample1.gather.with-lineages.csv`. This will produce an annotated gather CSV, `Sb47+63_gather_x_gtdbrs202_k31.with-lineages.csv`: ``` sourmash tax annotate --gather-csv Sb47+63_gather_x_gtdbrs202_k31.csv \ --taxonomy gtdb-rs202.taxonomy.v2.csv ``` The `with-lineages` output file format can be summarized with `sourmash tax summarize` and can also be used as an input taxonomy spreadsheet for any of the tax subcommands (new as of v4.6.0). ### `sourmash tax prepare` - prepare and/or combine taxonomy files `sourmash tax prepare` prepares taxonomy files for other `sourmash tax` commands. All `sourmash tax` commands must be given one or more taxonomy files as parameters to the `--taxonomy` argument. These files can be either CSV files or (as of sourmash 4.2.1) SQLite databases. SQLite databases are much faster for large taxonomies, while CSV files are easier to view and modify using spreadsheet software. `sourmash tax prepare` is a utility function that can ingest and validate multiple CSV files or SQLite databases, and output a CSV file or a SQLite database. It can be used to combine multiple taxonomies into a single file, as well as change formats between CSV and SQLite. The following command will take in two taxonomy files and combine them into a single taxonomy SQLite database. ``` sourmash tax prepare --taxonomy file1.csv file2.csv -o tax.db ``` Input databases formats can be mixed and matched, and the output format can be set to CSV like so: ``` sourmash tax prepare --taxonomy file1.csv file2.db -o tax.csv -F csv ``` **Note:** As of sourmash v4.6.0, the output of `sourmash tax annotate` can be used as a taxonomy input spreadsheet as well. ### `sourmash tax grep` - subset taxonomies and create picklists based on taxonomy string matches (`sourmash tax grep` is a new command as of sourmash v4.5.0.) `sourmash tax grep` searches taxonomies for matching strings, optionally restricting the string search to a specific taxonomic rank. It creates new files containing matching taxonomic entries; these new files can serve as taxonomies and can also be used as [picklists to restrict database matches](#using-picklists-to-subset-large-collections-of-signatures). Usage: ``` sourmash tax grep -t [ ...] ``` where `pattern` is a regular expression; see Python's [Regular Expression HOWTO for details on supported regexp features](https://docs.python.org/3/howto/regex.html#regex-howto). For example, ``` sourmash tax grep Shew -t gtdb-rs207.taxonomy.sqldb -o shew-picklist.csv ``` will search for a string match to `Shew` within the entire GTDB RS207 taxonomy, and will output a subset taxonomy in `shew-picklist.csv`. This picklist can be used with the GTDB RS207 databases like so: ``` sourmash search query.sig gtdb-rs207.genomic.k31.zip \ --picklist shew-picklist.csv:ident:ident ``` `tax grep` can also restrict string matching to a specific taxonomic rank with `-r/--rank`; for example, ``` sourmash tax grep Shew -t gtdb-rs207.taxonomy.sqldb \ -o shew-picklist.csv -r genus ``` will restrict matches to the rank of genus. Available ranks are superkingdom, phylum, class, order, family, genus, and species. `tax grep` also takes several standard grep arguments, including `-i` to ignore case and `-v` to output only taxonomic lineages that do _not_ match the pattern. Note: `tax grep` only searches taxonomic ranks, not identifier strings. Use `sig grep` to search for identifiers in sketch collections. Currently only CSV output (optionally gzipped) is supported; use `sourmash tax prepare` to convert CSV output from `tax grep` into a SQLite taxonomy database. ### `sourmash tax summarize` - print summary information for lineage spreadsheets or taxonomy databases (`sourmash tax summarize` is a new command as of sourmash v4.6.0.) `sourmash tax summarize` loads in one or more lineage spreadsheets, counts the distinct taxonomic lineages, and outputs a summary. It optionally will output a CSV file with a detailed count of how many identifiers belong to each taxonomic lineage. For example, ``` sourmash tax summarize gtdb-rs202.taxonomy.v2.db -o ranks.csv ``` outputs ``` number of distinct taxonomic lineages: 258406 rank superkingdom: 2 distinct taxonomic lineages rank phylum: 169 distinct taxonomic lineages rank class: 419 distinct taxonomic lineages rank order: 1312 distinct taxonomic lineages rank family: 3264 distinct taxonomic lineages rank genus: 12888 distinct taxonomic lineages rank species: 47894 distinct taxonomic lineages ``` and creates a file `ranks.csv` with the number of distinct identifier counts for each lineage at each rank: ``` rank,lineage_count,lineage superkingdom,254090,d__Bacteria phylum,120757,d__Bacteria;p__Proteobacteria class,104665,d__Bacteria;p__Proteobacteria;c__Gammaproteobacteria order,64157,d__Bacteria;p__Proteobacteria;c__Gammaproteobacteria;o__Enterobacterales family,55347,d__Bacteria;p__Proteobacteria;c__Gammaproteobacteria;o__Enterobacterales;f__Enterobacteriaceae ... ``` That is, there are 254,090 identifiers in GTDB rs202 under `d__Bacteria`, and 120,757 within the `p__Proteobacteria`. `tax summarize` can also be used to summarize the output of `tax annotate`. ## `sourmash lca` subcommands for in-memory taxonomy integration These commands use LCA databases (created with `lca index`, below, or prepared databases such as [genbank-k31.lca.json.gz](databases.md)). ### `sourmash lca classify` - classify a genome using an LCA database `sourmash lca classify` classifies one or more signatures using the given list of LCA DBs. It is meant for classifying metagenome-assembled genome bins (MAGs) and single-cell genomes (SAGs). ```{attention} We no longer recommend using `sourmash lca` for taxonomic analysis; please use `sourmash tax` instead. See [taxonomic profiling with sourmash](classifying-signatures.md#taxonomic-profiling-with-sourmash) for more information. ``` Usage: ``` sourmash lca classify --query query.sig [query2.sig ...] --db [ ...] ``` For example, the command ``` sourmash lca classify --query tests/test-data/63.fa.sig \ --db podar-ref.lca.json ``` will produce the following logging to stderr: ``` loaded 1 LCA databases. ksize=31, scaled=10000 finding query signatures... outputting classifications to stdout ... classifying NC_011663.1 Shewanella baltica OS223, complete genome classified 1 signatures total ``` and the example classification output is a CSV file with headers: ``` ID,status,superkingdom,phylum,class,order,family,genus,species "NC_009665.1 Shewanella baltica OS185, complete genome",found,Bacteria,Proteobacteria,Gammaproteobacteria,Alteromonadales,Shewanellaceae,Shewanella,Shewanella baltica ``` The `status` column in the classification output can take three possible values: `nomatch`, `found`, and `disagree`. `nomatch` means that no match was found for this query, and `found` means that an unambiguous assignment was found - all k-mers were classified within the same taxonomic hierarchy, and the most detailed lineage available was reported. `disagree` means that there was a taxonomic disagreement, and the lowest compatible taxonomic node was reported. To elaborate on this a bit, suppose that all of the k-mers within a signature were classified as family *Shewanellaceae*, genus *Shewanella*, or species *Shewanella baltica*. Then the lowest compatible node (here species *Shewanella baltica*) would be reported, and the status of the classification would be `found`. However, if a number of additional k-mers in the input signature were classified as *Shewanella oneidensis*, sourmash would be unable to resolve the taxonomic assignment below genus *Shewanella* and it would report a status of `disagree` with the genus-level assignment of *Shewanella*; species level assignments would not be reported. Here, the assigned rank is the rank immediately *above* where there is a taxonomic disagreement, and the taxid & lineage refer to the name at that rank (the lowest common ancestor at which an assignment can be made). For another example, if you saw this line in the CSV file: ``` TARA_ASW_MAG_00029,1224,disagree,phylum,Bacteria;Proteobacteria ``` you would know that TARA_ASW_MAG_00029 has k-mers that are shared between different orders: 'Pseudomonadales' and 'Rhodobacterales'. Therefore, the classifier status is `disagree`, and the classified taxid is at rank `phylum` - just above `order`. (This is the approach that Kraken and other lowest common ancestor implementations use, we believe.) Note: you can specify a list of file names to load signatures from in a text file passed to `sourmash lca classify` with the `--query-from-file` flag; these files will be appended to the `--query` input. ### `sourmash lca summarize` - summarize a metagenome's contents using an LCA database `sourmash lca summarize` produces a Kraken-style summary of the combined contents of the given query signatures. It is meant for exploring metagenomes and metagenome-assembled genome bins. `sourmash lca summarize` also weights output with hash abundances, so that output percentages are weighted by the number of times a k-mer is seen; this can be turned off with `--ignore-abundance`. ```{attention} We no longer recommend using `sourmash lca` for taxonomic analysis; please use `sourmash tax` instead. See [taxonomic profiling with sourmash](classifying-signatures.md#taxonomic-profiling-with-sourmash) for more information. ``` Usage: ``` sourmash lca summarize --query query.sig [query2.sig ...] --db [ ...] ``` For example, with the data in `tests/test-data/fake-abund`, the command line: ``` sourmash lca summarize --query query.sig.gz --db matches.lca.json.gz ``` will produce the following log output to stderr: ``` loaded 1 LCA databases. ksize=31, scaled=10000 finding query signatures... loaded 1 signatures from 1 files total. ``` and the following example summarize output to stdout: ``` 79.6% 550 Bacteria;Proteobacteria;Gammaproteobacteria;Alteromonadales;Shewanellaceae;Shewanella;Shewanella baltica;Shewanella baltica OS223 79.6% 550 Bacteria;Proteobacteria;Gammaproteobacteria;Alteromonadales;Shewanellaceae;Shewanella;Shewanella baltica 79.6% 550 Bacteria;Proteobacteria;Gammaproteobacteria;Alteromonadales;Shewanellaceae;Shewanella 79.6% 550 Bacteria;Proteobacteria;Gammaproteobacteria;Alteromonadales;Shewanellaceae 79.6% 550 Bacteria;Proteobacteria;Gammaproteobacteria;Alteromonadales 79.6% 550 Bacteria;Proteobacteria;Gammaproteobacteria 79.6% 550 Bacteria;Proteobacteria 79.6% 550 Bacteria 20.4% 141 Archaea;Euryarchaeota;unassigned;unassigned;unassigned;Aciduliprofundum;Aciduliprofundum boonei;Aciduliprofundum boonei T469 20.4% 141 Archaea;Euryarchaeota;unassigned;unassigned;unassigned;Aciduliprofundum;Aciduliprofundum boonei 20.4% 141 Archaea;Euryarchaeota;unassigned;unassigned;unassigned;Aciduliprofundum 20.4% 141 Archaea;Euryarchaeota;unassigned;unassigned;unassigned 20.4% 141 Archaea;Euryarchaeota;unassigned;unassigned 20.4% 141 Archaea;Euryarchaeota;unassigned 20.4% 141 Archaea;Euryarchaeota 20.4% 141 Archaea ``` The output is space-separated and consists of three columns: the percentage of total k-mers that have this classification; the number of k-mers that have this classification; and the lineage classification. K-mer classifications are reported hierarchically, so the percentages and totals contain all assignments that are at a lower taxonomic level - e.g. *Bacteria*, above, contains all the k-mers in *Bacteria;Proteobacteria*. The same information is reported in a CSV file if `-o/--output` is used. The proportions reflect the query signature construction, where the metagenome contains a 1.5 Mbp Archaeal genome and a 5.4 Mbp Bacterial genome. The Archaeal genome is therefore only ~20% of the distinct k-mers in the metagenome (1.5 Mbp divided by 6.9 Mbp). If `--with-abundance` is given, the output changes to reflect the proportions of the query metagenome based on k-mer/read abundances: ``` 56.8% 740 Archaea;Euryarchaeota;unassigned;unassigned;unassigned;Aciduliprofundum;Aciduliprofundum boonei;Aciduliprofundum boonei T469 56.8% 740 Archaea;Euryarchaeota;unassigned;unassigned;unassigned;Aciduliprofundum;Aciduliprofundum boonei 56.8% 740 Archaea;Euryarchaeota;unassigned;unassigned;unassigned;Aciduliprofundum 56.8% 740 Archaea;Euryarchaeota;unassigned;unassigned;unassigned 56.8% 740 Archaea;Euryarchaeota;unassigned;unassigned 56.8% 740 Archaea;Euryarchaeota;unassigned 56.8% 740 Archaea;Euryarchaeota 56.8% 740 Archaea 43.2% 563 Bacteria;Proteobacteria;Gammaproteobacteria;Alteromonadales;Shewanellaceae;Shewanella;Shewanella baltica;Shewanella baltica OS223 43.2% 563 Bacteria;Proteobacteria;Gammaproteobacteria;Alteromonadales;Shewanellaceae;Shewanella;Shewanella baltica 43.2% 563 Bacteria;Proteobacteria;Gammaproteobacteria;Alteromonadales;Shewanellaceae;Shewanella 43.2% 563 Bacteria;Proteobacteria;Gammaproteobacteria;Alteromonadales;Shewanellaceae 43.2% 563 Bacteria;Proteobacteria;Gammaproteobacteria;Alteromonadales 43.2% 563 Bacteria;Proteobacteria;Gammaproteobacteria 43.2% 563 Bacteria;Proteobacteria 43.2% 563 Bacteria ``` Here, the changed proportions reflect the query signature abundances, where the 1.5 Mbp Archaeal genome is present 5 times, while the 5.4 Mbp Bacterial genome is present only once; when weighted by abundance, the Bacterial genome is only 41.8% of the metagenome content, while the Archaeal genome is 58.1% of the metagenome content. Note: you can specify a list of file names to load signatures from in a text file passed to `sourmash lca summarize` with the `--query-from-file` flag; these files will be appended to the `--query` input. ### `sourmash lca index` - build an LCA database The `sourmash lca index` command creates an LCA database from a lineage spreadsheet and a collection of signatures. This can be used to create LCA databases from private collections of genomes, and can also be used to create databases for e.g. subsets of GenBank. See [the `sourmash lca` tutorial](tutorials-lca.md) and the blog post [Why are taxonomic assignments so different for Tara bins?](http://ivory.idyll.org/blog/2017-taxonomic-disagreements-in-tara-mags.html) for some use cases. If you are interested in preparing lineage spreadsheets from GenBank genomes (or building off of NCBI taxonomies more generally), please see [the NCBI lineage repository](https://github.com/dib-lab/2018-ncbi-lineages). You can use `--from-file` to pass `lca index` a text file containing a list of file names to index. As of sourmash 4.2.0, `lca index` supports `--picklist`, to [select a subset of signatures based on a CSV file](#using-picklists-to-subset-large-collections-of-signatures). This can be used to index a subset of a large collection, or to exclude a few signatures from an index being built from a large collection. As of sourmash 4.4.0, `lca index` can produce an _on disk_ LCA database using SQLite. To prepare such a database, use `sourmash lca index ... -F sql`. All sourmash commands work with either type of LCA database (the default JSON database, and the SQLite version). SQLite databases are larger than JSON databases on disk but are typically much faster to load and search, and use much less memory. ### `sourmash lca rankinfo` - examine an LCA database The `sourmash lca rankinfo` command displays k-mer specificity information for one or more LCA databases. See the blog post [How specific are k-mers for taxonomic assignment of microbes, anyway?](http://ivory.idyll.org/blog/2017-how-specific-kmers.html) for example output. ### `sourmash lca compare_csv` - compare taxonomic spreadsheets The `sourmash lca compare_csv` command compares two lineage spreadsheets (such as those output by `sourmash lca classify` or taken as input by `sourmash lca index`) and summarizes their agreement/disagreement. Please see the blog post [Why are taxonomic assignments so different for Tara bins?](http://ivory.idyll.org/blog/2017-taxonomic-disagreements-in-tara-mags.html) for an example use case. [0]:https://www.biorxiv.org/content/10.1101/2022.01.11.475838v2 [1]:http://mash.readthedocs.io/en/latest/ [2]:http://biorxiv.org/content/early/2015/10/26/029827 [3]:https://en.wikipedia.org/wiki/Jaccard_index ## `sourmash signature` subcommands for signature manipulation These commands manipulate signatures from the command line. The signature commands that combine or otherwise have multiple signatures interacting (`merge`, `intersect`, `subtract`) work only on compatible signatures, where the k-mer size and nucleotide/protein sequences match each other. If working directly with the hash values (e.g. `merge`, `intersect`, `subtract`) then the scaled values must also match; you can use `downsample` to convert a bunch of samples to the same scaled value. If there are multiple signatures in a file with different ksizes and/or from nucleotide and protein sequences, you can choose amongst them with `-k/--ksize` and `--dna/--protein/--hp/--dayhoff/--skipm1n3/--skipm2n3`, as with other sourmash commands such as `search`, `gather`, and `compare`. Note, you can use `sourmash sig` as shorthand for all of these commands. All commands load signatures in all supported sourmash formats, including indexed databases, and will save signatures in formats based on the extension (e.g. `-o output.zip` will use the zip format). You can load signatures from stdin using `-` on the command line. ### `sourmash signature cat` - combine signatures into one file Concatenate signature files. For example, ``` sourmash signature cat file1.sig file2.sig -o all.sig.zip ``` will combine all signatures in `file1.sig` and `file2.sig` and put them in the file `all.sig.zip`. #### Using picklists with `sourmash sig cat` As of sourmash 4.2.0, `cat` also supports picklists, a feature by which you can select signatures based on values in a CSV file. See [Using picklists to subset large collections of signatures](#using-picklists-to-subset-large-collections-of-signatures), below. ### `sourmash signature describe` - display detailed information about signatures Display signature details. For example, ``` sourmash sig describe tests/test-data/track_abund/47.fa.sig ``` will display: ``` signature filename: tests/test-data/track_abund/47.fa.sig signature: NC_009665.1 Shewanella baltica OS185, complete genome source file: podar-ref/47.fa md5: 09a08691ce52952152f0e866a59f6261 k=31 molecule=DNA num=0 scaled=1000 seed=42 track_abundance=1 size: 5177 sum hashes: 5292 signature license: CC0 ``` Here, the `size` is the number of distinct hashes in the sketch, and `sum_hashes` is the total number of hashes in the sketch, with abundances. When `track_abundance` is 0, `size` is always the same as `sum_hashes`. ### `sourmash signature fileinfo` - display a summary of the contents of a sourmash collection Display signature file, database, or collection. For example, ``` sourmash sig fileinfo tests/test-data/prot/all.zip ``` will display: ``` path filetype: ZipFileLinearIndex location: /Users/t/dev/sourmash/tests/test-data/prot/all.zip is database? yes has manifest? yes is nonempty? yes num signatures: 8 ** examining manifest... 31758 total hashes summary of sketches: 2 sketches with dayhoff, k=19, scaled=100 7945 total hashes 2 sketches with hp, k=19, scaled=100 5184 total hashes 2 sketches with protein, k=19, scaled=100 8214 total hashes 2 sketches with DNA, k=31, scaled=1000 10415 total hashes ``` `sig fileinfo` will recognize [all accepted sourmash input files](#loading-signatures-and-databases), including individual .sig and .sig.gz files, Zip file collections, SBT databases, LCA databases, and directory hierarchies. `sourmash sig fileinfo` provides optional JSON and YAML output, and those formats are under semantic versioning. Note: `sourmash signature summarize` is an alias for `fileinfo`; they are the same command. ### `sourmash signature grep` - extract matching signatures using pattern matching Extract matching signatures with substring and regular expression matching on the name, filename, and md5 fields. For example, ``` sourmash signature grep -i shewanella tests/test-data/prot/all.zip -o shew.sig.zip ``` will extract the two signatures in `all.zip` with 'Shewanella baltica' in their name and save them to `shew.sig.zip`. `grep` will search for substring matches or regular expressions; e.g. `sourmash sig grep 'os185|os223' ...` will find matches to either of those expressions. String matching modifiers include `-i` for case-insensitive matching, and `-v` for exclusion rather than inclusion. By default, `sig grep` outputs matching sketches. Alternatively, `-l/--print-matched-names` prints a list of distinct matching sketch names and `-c/--count` prints a count of total matching sketches; both disable sketch output by setting `--no-sigs`. A CSV file of the matching sketch information can be saved using `--csv `; this file is in the sourmash manifest format and can be used as a picklist with `--pickfile ::manifest`. If `--no-sigs` is specified, `sourmash sig grep` will not output matching signatures. ### `sourmash signature split` - split signatures into individual files Split each signature in the input file(s) into individual files, with standardized names. For example, ``` sourmash signature split tests/test-data/2.fa.sig ``` will create 3 files, `f372e478.k=21.scaled=1000.DNA.dup=0.2.fa.sig`, `f3a90d4e.k=31.scaled=1000.DNA.dup=0.2.fa.sig`, and `43f3b48e.k=51.scaled=1000.DNA.dup=0.2.fa.sig`, representing the three different DNA signatures at different ksizes created from the input file `2.fa`. The format of the names of the output files is standardized and stable for major versions of sourmash: currently, they are period-separated with fields: * `md5sum` - a unique hash value based on the contents of the signature. * `k=` - k-mer size. * `scaled=` or `num=` - scaled or num value for MinHash. * `` - the molecule type (DNA, protein, dayhoff, hp, skipm1n3, or skipm2n3) * `dup=` - a non-negative integer that prevents duplicate signatures from colliding. * `basename` - basename of first input file used to create signature; if none provided, or stdin, this is `none`. If `--outdir` is specified, all of the signatures are placed in outdir. Note: `split` only saves files in the JSON `.sig` format. ### `sourmash signature merge` - merge two or more signatures into one Merge two (or more) signatures. For example, ``` sourmash signature merge file1.sig file2.sig -o merged.sig ``` will output the union of all the hashes in `file1.sig` and `file2.sig` to `merged.sig`. All of the signatures passed to merge must either have been created with `-p abund`, or not. If they have `track_abundance` on, then the merged signature will have the sum of all abundances across the individual signatures. The `--flatten` flag will override this behavior and allow merging of mixtures by removing all abundances. `sig merge` can only merge compatible sketches - if there are multiple k-mer sizes or molecule types present in any of the signature files, you will need to choose one k-mer size with `-k/--ksize`, and/or one moltype with `--dna/--protein/--hp/--dayhoff/--skipm1n3/--skipm2n3`. Use `--set-name ` to set the name of the output sketch. Note: `merge` only creates one output file, with one signature in it. ### `sourmash signature rename` - rename a signature Rename the display name for one or more signatures - this is the name output for matches in `compare`, `search`, `gather`, etc. For example, ``` sourmash signature rename file1.sig "new name" -o renamed.sig ``` will place a renamed copy of the hashes in `file1.sig` in the file `renamed.sig`. If you provide multiple signatures, all will be renamed to the same name. ### `sourmash signature subtract` - subtract other signatures from a signature Subtract all of the hash values from one signature that are in one or more of the others. For example, ``` sourmash signature subtract file1.sig file2.sig file3.sig -o subtracted.sig ``` will subtract all of the hashes in `file2.sig` and `file3.sig` from `file1.sig`, and save the new signature to `subtracted.sig`. To use `subtract` on signatures calculated with `-p abund`, you must specify `--flatten`. `sig subtract` can only work with compatible sketches - if there are multiple k-mer sizes or molecule types present in any of the signature files, you will need to choose one k-mer size with `-k/--ksize`, and/or one moltype with `--dna/--protein/--hp/--dayhoff/--skipm1n3/--skipm2n3`. Use `--set-name ` to set the name of the output sketch. Note: `subtract` only creates one output file, with one signature in it. ### `sourmash signature intersect` - intersect two (or more) signatures Output the intersection of the hash values in multiple signature files. For example, ``` sourmash signature intersect file1.sig file2.sig file3.sig -o intersect.sig ``` will output the intersection of all the hashes in those three files to `intersect.sig`. The `intersect` command flattens all signatures, i.e. the abundances in any signatures will be ignored and the output signature will have `track_abundance` turned off. The `-A/--abundance-from` argument will borrow abundances from the specified signature (which will also be added to the intersection). `sig intersect` can only work with compatible sketches - if there are multiple k-mer sizes or molecule types present in any of the signature files, you will need to choose one k-mer size with `-k/--ksize`, and/or one moltype with `--dna/--protein/--hp/--dayhoff/--skipm1n3/--skipm2n3`. Use `--set-name ` to set the name of the output sketch(es). ### `sourmash signature inflate` - transfer abundances from one signature to others Use abundances from one signature to provide abundances on other signatures. For example, ``` sourmash signature inflate file1.sig file2.sig file3.sig -o inflated.sig ``` will take the abundances from hashes `file1.sig` and use them to set the abundances on matching hashes in `file2.sig` and `file3.sig`. Any hashes that are not present in `file1.sig` will be removed from `file2.sig` and `file3.sig` as they will now have zero abundance. `sig inflate` can only work with compatible sketches - if there are multiple k-mer sizes or molecule types present in any of the signature files, you will need to choose one k-mer size with `-k/--ksize`, and/or one moltype with `--dna/--protein/--hp/--dayhoff/--skipm1n3/--skipm2n3`. ### `sourmash signature downsample` - decrease the size of a signature Downsample one or more signatures. With `downsample`, you can -- * increase the `scaled` value for a signature created with `-p scaled=SCALED`, shrinking it in size; * decrease the `num` value for a traditional num MinHash, shrinking it in size; * try to convert a `scaled` signature to a `num` signature; * try to convert a `num` signature to a `scaled` signature. For example, ``` sourmash signature downsample file1.sig file2.sig --scaled 100000 -o downsampled.sig ``` will output each signature, downsampled to a scaled value of 100000, to `downsampled.sig`; and ``` sourmash signature downsample --num 500 scaled_file.sig -o downsampled.sig ``` will try to convert a scaled MinHash to a num MinHash. ### `sourmash signature extract` - extract signatures from a collection Extract the specified signature(s) from a collection of signatures. For example, ``` sourmash signature extract *.sig -k 21 --dna -o extracted.sig ``` will extract all nucleotide signatures calculated at k=21 from all .sig files in the current directory. There are currently two other useful selectors for `extract`: you can specify (part of) an md5sum, as output in the CSVs produced by `search` and `gather`; and you can specify (part of) a name. For example, ``` sourmash signature extract tests/test-data/*.fa.sig --md5 09a0869 ``` will extract the signature from `47.fa.sig` which has an md5sum of `09a08691ce52952152f0e866a59f6261`; and ``` sourmash signature extract tests/test-data/*.fa.sig --name NC_009665 ``` will extract the same signature, which has an accession number of `NC_009665.1`. #### Using picklists with `sourmash sig extract` As of sourmash 4.2.0, `extract` also supports picklists, a feature by which you can select signatures based on values in a CSV file. See [Using picklists to subset large collections of signatures](#using-picklists-to-subset-large-collections-of-signatures), below. ### `sourmash signature flatten` - remove abundance information from signatures Flatten the specified signature(s), removing abundances and setting track_abundance to False. For example, ``` sourmash signature flatten *.sig -o flattened.sig ``` will remove all abundances from all of the .sig files in the current directory. The `flatten` command accepts the same selectors as `extract`. ### `sourmash signature filter` - remove hashes based on abundance Filter the hashes in the specified signature(s) by abundance, by either `-m/--min-abundance` or `-M/--max-abundance` or both. Abundance selection is inclusive, so `-m 2 -M 5` will select hashes with abundance greater than or equal to 2, and less than or equal to 5. For example, ``` sourmash signature filter -m 2 *.sig ``` will output new signatures containing only hashes that occur two or more times in each signature. The `filter` command accepts the same selectors as `extract`. ### `sourmash signature import` - import signatures from mash. Import signatures into sourmash format. Currently only supports mash, and can import mash sketches output by `mash info -d `. For example, ``` sourmash signature import filename.msh.json -o imported.sig ``` will import the contents of `filename.msh.json` into `imported.sig`. Note: `import` only creates one output file, with one signature in it. Note: `ingest` is an alias for `import`. ### `sourmash signature export` - export signatures to mash. Export signatures from sourmash format. Currently only supports mash dump format. For example, ``` sourmash signature export filename.sig -o filename.sig.msh.json ``` ### `sourmash signature overlap` - detailed comparison of two signatures' overlap Display a detailed comparison of two signatures. This calculates the Jaccard similarity (as in `sourmash compare` or `sourmash search`) and the Jaccard containment in both directions (as with `--containment`). It also displays the number of hash values in the union and intersection of the two signatures, as well as the number of disjoint hash values in each signature. This command has two uses - first, it is helpful for understanding how similarity and containment are calculated, and second, it is useful for analyzing signatures with very small overlaps, where the similarity and/or containment might be very close to zero. For example, ``` sourmash signature overlap tests/test-data/63.fa.sig \ tests/test-data/47.fa.sig ``` will display the detailed comparison of the two files like so: ```text loaded one signature each from tests/test-data/63.fa.sig and tests/test-data/47.fa.sig first signature: signature filename: tests/test-data/63.fa.sig signature: NC_011663.1 Shewanella baltica OS223, complete genome md5: 38729c6374925585db28916b82a6f513 k=31 molecule=DNA num=0 scaled=1000 second signature: signature filename: tests/test-data/47.fa.sig signature: NC_009665.1 Shewanella baltica OS185, complete genome md5: 09a08691ce52952152f0e866a59f6261 k=31 molecule=DNA num=0 scaled=1000 similarity: 0.32069 first contained in second: 0.48282 second contained in first: 0.48851 number of hashes in first: 5238 number of hashes in second: 5177 number of hashes in common: 2529 only in first: 2709 only in second: 2648 total (union): 7886 ``` `sig overlap` can only work with compatible sketches - if there are multiple k-mer sizes or molecule types present in any of the signature files, you will need to choose one k-mer size with `-k/--ksize`, and/or one moltype with `--dna/--protein/--hp/--dayhoff/--skipm1n3/--skipm2n3`. ### `sourmash signature kmers` - extract k-mers and/or sequences that match to signatures Given one or more compatible sketches and some sequence files, extract the k-mers and/or sequences corresponding to the hash values in the sketch. Because the sourmash hash function is one-way, this requires FASTA or FASTQ sequence files in addition to the sketch. For example, ``` sourmash sig kmers --signatures sig1.sig --sequences seqfile.fasta \ --save-sequences matches.fasta --save-kmers kmer-matches.csv ``` will search `seqfile.fasta` for matching sequences and k-mers, and produce two files. The file `matches.fasta` will contain FASTA sequences that match the hashes in the input signature, while the file `kmer-matches.csv` provides the matching k-mers and hash values, together with their originating filename and sequence name. If the sketch is a protein sketch (protein, dayhoff, or hp), then the input sequences are assumed to be protein. To search DNA sequences for translated protein hashes, provide the `--translate` flag to `sig kmers`. `--save-sequences` and `--save-kmers` are both optional. If neither are given, basic statistics on k-mer matching are given. Please note that `--save-kmers` can be very slow on large files! The input sketches are the source of the input hashes. So, for example, If `--scaled=1` sketches are provided, `sig kmers` can be used to yield all the k-mers and their matching hashes. Likewise, if the sketch is built from the intersection of two other sketches, only the k-mers and hash values present in both sketches will be used. Likewise, the input sequences are used for matching; they do not need to be the same sequences that were used to create the sketches. Input sequences can be in FASTA or FASTQ format, and either flat text or compressed with gzip or bzip2; formats are auto-detected. By default, `sig kmers` ignores bad k-mers (e.g. non-ACGT characters in DNA). If `--check-sequence` is provided, `sig kmers` will error exit on the first bad k-mer. If `--check-sequence --force` is provided, `sig kmers` will provide error messages (and skip bad sequences), but will continue processing input sequences. ### `sourmash signature manifest` - output a manifest for a file Output a manifest for a file, database, or collection. Note that these manifests are not usually suitable for use as standalone manifests; the `sourmash sig collect` and `sourmash sig check` commands produce standalone manifests. For example, ``` sourmash sig manifest tests/test-data/prot/all.zip -o manifest.csv ``` will create a CSV file, `manifest.csv`, in the internal sourmash manifest format. The manifest will contain an entry for every signature in the file, database, or collection. This format is largely meant for internal use, but it can serve as a [picklist pickfile](#using-picklists-to-subset-large-collections-of-signatures) for subsetting large collections. By default, `sourmash sig manifest` will rebuild the manifest by iterating over the signatures in the input file. This can be slow for large collections. Use `--no-rebuild-manifest` to load an existing manifest if it is available. As of sourmash 4.4.0, `sig manifest` can produce a manifest in a fast on-disk format (a SQLite database). SQLite manifests can be _much_ faster when working with very large collections of signatures. To produce a SQLite manifest, use `sourmash sig manifest ... -F sql`. All sourmash commands that work with manifests will accept both CSV and SQLite manifest files. :::{warning} **sourmash v5 breaking change:** In sourmash v5.0 and later, `--no-rebuild-manifest` will become default. You can specify `--v4` to keep the old behavior, or `--v5` to support the new default behavior, or simply specify `--no-rebuild-manifest` to override the default. See [Migrating from sourmash v4.x to sourmash v5.x](support.md#migrating-from-sourmash-v4x-to-sourmash-v5x) for more details. ::: ### `sourmash signature check` - compare picklists and manifests Compare picklists and manifests across databases, and optionally output matches and missing items. In particular, `sig check` can be used to create standalone manifests for a subset of a large collection, using picklists. For example, ``` sourmash sig check tests/test-data/gather/GCF*.sig \ --picklist tests/test-data/gather/salmonella-picklist.csv::manifest ``` will load all of the `GCF` signatures and compare them to the given picklist. With `-o/--output-missing`, `sig check` will save unmatched elements of the picklist CSV. With `--save-manifest-matching`, `sig check` will save all of the _matched_ elements to a manifest file, which can then be used as a sourmash database. `sourmash sig check` is particularly useful when working with large collections of signatures and identifiers. With `-m/--save-manifest-matching`, `sig check` creates a standalone manifest. In these manifests, sourmash v4 will by default write paths to the matched elements that are relative to the current working directory. In some cases - when the output manifest is in a different directory - this will create manifests that do not work properly with sourmash. The `--relpath` argument will rewrite the paths to be relative to the manifest, while the `--abspath` argument will rewrite paths to be absolute. The `--relpath` behavior will be the default in sourmash v5. Standalone manifests created with `-m/--save-manifest-matching` will use the paths given to `sig check` on the command line; we recommend using zip files and sig files, and avoiding directory hierarchies or path lists. You can use `--from-file` to pass in long lists of filenames via a text file. :::{warning} **sourmash v5 breaking change:** In sourmash v5.0 and later, `--relpath` will become default. You can specify `--v4` to keep the old behavior, or `--v5` to support the new default behavior, or simply specify `--relpath` or `--abspath` to specify the desired behavior. See [Migrating from sourmash v4.x to sourmash v5.x](support.md#migrating-from-sourmash-v4x-to-sourmash-v5x) for more details. ::: ### `sourmash signature collect` - collect manifests across databases Collect manifests from across (many) files and merge into a single standalone manifest. Standalone manifests can be used directly as a sourmash database; they support efficient searching and selection of sketches, as well as lazy loading of individual sketches from large collections. See [advanced usage information on sourmash databases](databases-advanced.md) for more information. For example, ``` sourmash sig collect tests/test-data/gather/GCF*.sig -o mf.sqlmf ``` will load all of the `GCF` signatures and build a manifest file `mf.sqlmf` that contains references to all of the signatures, but not the signatures themselves. This manifest file can be loaded directly from the command line by sourmash. `sourmash sig collect` defaults to outputting SQLite manifests. It is particularly useful when working with large collections of signatures and identifiers, and has command line options for merging and updating manifests. The standalone manifests created by `sig collect` will reference the paths given on the command line; we recommend using zip files and sig files, and avoiding directory hierarchies or path lists. You can also use `--from-file` to pass in long lists of filenames. Standalone manifests produced by `sig collect` work most efficiently when constructed from many small zip file collections. As with `sig check`, the standalone manifests created by `sig collect` in sourmash v4 will by default write paths to the matched elements relative to the current working directory. When the output manifest is in a different directory, this will create manifests that do not work properly with sourmash. The `--relpath` argument will rewrite the paths to be relative to the manifest, while the `--abspath` argument will rewrite paths to be absolute. The `--relpath` behavior will be the default in sourmash v5. ## Advanced command-line usage ### Loading signatures and databases sourmash uses several different command-line styles. Most sourmash commands can load sketches from any standard collection type; we primarily recommend using zipfiles (but read on!) Briefly, * `search` and `gather` both take a single query signature and search multiple signatures or databases. In this case, there has to be a single identifiable query for sourmash to use, and if you're using a database or list of signatures as the source of a query, you'll need to provide a selector (ksize with `-k`, moltype with `--dna/--protein/--hp/--dayhoff/--skipm1n3/--skipm2n3` etc, or md5sum with `--query-md5`) that picks out a single signature. * `compare` takes multiple signatures and can load them from any sourmash collection type. * the `lca classify` and `lca summarize` commands take multiple signatures with `--query`, and multiple LCA databases, with `--db`. `sourmash multigather` also uses this style. This allows these commands to specify multiple queries **and** multiple databases without (too much) confusion. The database must be LCA databases. * `index` and `lca index` take a few fixed parameters (database name, and for `lca index`, a taxonomy file) and then an arbitrary number of other files that contain signatures. None of these commands currently support searching, comparing, or indexing signatures with multiple ksizes or moltypes at the same time; you need to pick the ksize and moltype to use for your query. Where possible, scaled values will be made compatible. ### Selecting signatures (sourmash v4.3.0 and later) sourmash is built to work with very large collections of signatures, and you may want to select (or exclude) specific signatures from search or other operations, based on their name. This can be done without modifying the collections themselves via the `--include-db-pattern` and `--exclude-db-pattern` arguments to many sourmash commands, including `search`, `gather`, `compare`, `prefetch`, and `sig extract`. In brief, `sourmash search ... --include ` will search only those database signatures that match `` in their `name`, `filename`, or `md5` strings. Here, `` can be either a substring or a regular expression. Likewise, `sourmash search ... --exclude ` will search only those database signatures that _don't_ match pattern in their `name`, `filename`, or `md5` strings. ### Using picklists to subset large collections of signatures (sourmash v4.2.0 and later) Many commands support *picklists*, a feature by which you can select or "pick out" signatures based on values in a CSV file. This is typically used to index, extract, or search a subset of a large collection where modifying the collection itself isn't desired. For example, ``` sourmash sig extract --picklist list.csv:md5:md5sum ``` will extract only the signatures that have md5sums matching the column `md5sum` in the CSV file `list.csv`. The command ``` sourmash sig extract --picklist list.csv::prefetch ``` will extract only the signatures found in the output of `sourmash prefetch ... -o list.csv`. The `--picklist` argument string must be of the format `pickfile:colname:coltype[:pickstyle]`, where `pickfile` is the path to a CSV file, `colname` is the name of the column to select from the CSV file (based on the headers in the first line of the CSV file), and `coltype` is the type of match. An optional pickstyle argument, `:include` or `:exclude`, can be added as a fourth parameter; if omitted, the default is `:include`. The following `coltype`s are currently supported for picklists: * `name` - exact match to signature's name * `md5` - exact match to signature's md5sum * `md5prefix8` - match to 8-character prefix of signature's md5sum * `md5short` - same as `md5prefix8` * `ident` - exact match to signature's identifier * `identprefix` - match to signature's identifier, before '.' * `gather` - use the CSV output of `sourmash gather` as a picklist * `prefetch` - use the CSV output of `sourmash prefetch` as a picklist * `search` - use the CSV output of `sourmash prefetch` as a picklist * `manifest` - use CSV manifests produced by `sig manifest`, `sig collect`, or `sig check` as a picklist Identifiers are constructed by using the first space delimited word in the signature name. One way to build a picklist is to use `sourmash sig grep --csv out.csv` to construct a CSV file containing a list of all sketches that match the pattern (which can be a string or regexp). The `out.csv` file can be used as a picklist via the picklist manifest CSV format with `--picklist out.csv::manifest`. You can also use `sourmash sig describe --csv out.csv ` or `sourmash sig manifest -o out.csv ` to construct an initial CSV file that you can then edit further and use as a picklist as above. The picklist functionality also supports excluding (rather than including) signatures matching the picklist arguments. To specify a picklist for exclusion, add `:exclude` to the `--picklist` argument string, e.g. `pickfile:colname:coltype:exclude`. For example, ``` sourmash sig extract --picklist list.csv:md5:md5sum:exclude ``` will extract only the signatures that have md5sums that **do not** match entries in the column `md5sum` in the CSV file `list.csv`. In addition to `sig extract`, the following commands support `--picklist` selection: `index`, `search`, `gather`, `prefetch`, `compare`, `index`, and `lca index`. ### Storing (and searching) signatures Backing up a little, there are many ways to store and search signatures. `sourmash` supports storing and loading signatures from JSON files, directories, lists of files, Zip files, custom indexed databases, and SQLite databases. These can all be used interchangeably for most sourmash operations. The simplest is one signature in a single JSON file. You can also put many signatures in a single JSON file, either by building them that way with `sourmash sketch` or by using `sourmash sig cat` or other commands. Searching or comparing these files involves loading them sequentially and iterating across all of the signatures - which can be slow, especially for many (100s or 1000s) of signatures. ### Zip files All of the `sourmash` commands support loading collections of signatures from zip files. You can create a compressed collection of signatures using `sourmash sig cat *.sig -o collections.sig.zip` and then specifying `collections.sig.zip` on the command line in place of `*.sig`; you can also sketch FASTA/FASTQ files directly into a zip file with `-o collections.sig.zip`. ### Choosing signature output formats (sourmash v4.1 and later) All signature saving arguments (`--save-matches` for `search` and `gather`, `-o` for `sourmash sketch`, and `-o` for the `sourmash signature` commands) support flexible saving of collections of signatures into JSON text, Zip files, and/or directories. This behavior is triggered by the requested output filename -- * to save to JSON signature files, use `.sig`; using the filename `-` will send JSON to stdout. * to save to gzipped JSON signature files, use `.sig.gz`; * to save to a Zip file collection, use `.zip`; * to save signature files to a directory, use a name ending in `/`; the directory will be created if it doesn't exist; * to save to a SQLite database, use `.sqldb` (as of sourmash v4.4.0). If none of these file extensions is detected, output will be written in the JSON `.sig` format, either to the provided output filename or to stdout. All of these save formats can be loaded by sourmash commands. **We strongly suggest using .zip files to store signatures: they are fast, small, and fully supported by all the sourmash commands and API.** Note that when outputting large collections of signatures, some save formats require holding all the sketches in memory until they can be written out, and others can save progressively. This can affect memory usage! Currently `.sig` and `.sig.gz` formats are held in memory, while `.zip`, directory outputs, and `.sqldb` formats write progressively to disk. For more detailed information on database formats and performance tradeoffs, please see [the advanced usage information for databases!](databases-advanced.md) ### Loading many signatures #### Indexed databases Indexed databases can make searching signatures much, much faster. RocksDB databases produced by `sourmash index` are low memory and disk-intensive databases that allow for fast searches using an inverted index. (We continue to support other indexed database types, including SBTs, LCA databases, and SQLite databases, but recommend using RocksDB over these other formats.) Commands that take multiple signatures or collections of signatures will also work with one or more indexed databases. One limitation of indexed databases is that they are all restricted in to certain kinds of signatures. RocksDB, SBT, and LCA databases can only contain one "type" of signature (one ksize/one moltype at one scaled value). SQLite databases can contain multiple ksizes and moltypes, but only at one scaled value. If the database signature type is incompatible with the other signatures, sourmash will complain appropriately. In contrast, signature files and zip collections can contain many different types of signatures, and compatible ones will be selected automatically. Use the `sourmash index -F rocksdb` command to create a RocksDB index. #### Loading signatures within a directory hierarchy All of the `sourmash` commands support loading signatures (`.sig` or `.sig.gz` files) from within directory hierarchies; you can provide the paths to the top-level directory on the command line. However, this is no longer recommended because it can be very inefficient; we instead suggest passing all of the sketch files in the directory into `sig collect` to build a standalone manifest, or using `sig cat` on the directory to generate a zip file. #### Passing in lists of files sourmash commands support `--from-file` or `--query-from-file`, which will take the location of a text file containing a list of file paths. This can be useful for situations where you want to specify thousands of queries, or a subset of signatures produced by some other command. This is no longer recommended when using large collections; we instead suggest using standalone manifests built with `sig collect` and `sig check`, which will include extra metadata that supports fast loading. ### Combining search databases on the command line All of the commands in sourmash operate in "online" mode, so **you can combine multiple databases and signatures on the command line and get the same answer as if you built a single large database from all of them**. The only caveat to this rule is that if you have multiple identical matches present across the databases, the order in which they are used may depend on the order that the files are passed in on the command line. ### Using stdin Most commands will take signature JSON data via stdin using the usual UNIX convention, `-`. Moreover, `sourmash sketch` and the `sourmash sig` commands will output to stdout. So, for example, ``` sourmash sketch ... -o - | sourmash sig describe - ``` will describe the signatures that were just created. ### Using standalone manifests to explicitly refer to collections of files (sourmash v4.4 and later) Manifests are metadata catalogs of signatures that are used for signature selection and loading. They are used extensively by sourmash internals to speed up signature selection through picklists and pattern matching. Manifests can _also_ be used externally (via the command-line), and these "standalone manifests" may be useful for organizing large collections of signatures. They can be generated with the `sig collect`, `sig manifest`, and `sig check` subcommands. The only difference between standalone manifests and internal manifests are that standalone manifests contain externally resolvable path names in the `internal_location` column. Suppose you have a large collection of signatures (`.sig` or `.sig.gz` files) in a location (e.g., under a directory, or in a zip file). You can create a manifest file for them like so: ``` sourmash sig collect -o manifest.sqlmf ``` and then use the manifest directly for sourmash operations, for example: ``` sourmash sig fileinfo manifest.sqlmf ``` This manifest contains _references_ to the signatures (but not the signatures themselves) and can then be used as a database target for most sourmash operations - search, gather, etc. Manifests support fast selection and lazy loading of sketches in many situations. The `sig check` command can also be used to create standalone manifests from collections using a picklist, with the `-m/--save-manifest-matching` option. This is useful for commands that don't support picklists natively, such as commands in plugins. Note that `sig collect` and `sig check` will generate manifests containing the pathnames given to them - so if you use relative paths, the references will be relative to the working directory in which the command was run. You can use `sig collect --abspath` to rewrite the paths into absolute paths, or `sig collect --relpath` to rewrite the paths relative to the manifest file. **Our advice:** We suggest using zip file collections for most situations; we strongly recommend using standalone manifests for situations where you have **very large** sketches or a **very large** collection of sketches (1000s or more), and don't want to make multiple copies of signatures in the collection (as you would have to, with a zipfile). This is particularly useful if you want to refer to different subsets of the collection without making multiple copies in a zip file. You can read more about the details of zip files and manifests in [the advanced usage information for databases](databases-advanced.md). ## Using sourmash plugins As of sourmash v4.7.0, sourmash has an experimental plugins interface! The plugin interface supports extending sourmash to load and save signatures in new ways, and also supports the addition of sourmash subcommands via `sourmash scripts`. In order to use a plugin with sourmash, you will need to use `pip` or `conda` to install the plugin the same environment that sourmash is installed in. In the future, we will include a list of available sourmash plugins in the documentation, and also provide a way to list available plugins. You can list all installed plugins and their versions with `sourmash info -v`. Below are some useful plugins that the sourmash team uses regularly and supports! ### The `branchwater` plugin - multithreaded and optimized sourmash operations (Installable via conda and pip as `sourmash_plugin_branchwater`.) The [`branchwater` plugin](https://github.com/sourmash-bio/sourmash_plugin_branchwater) provides faster and lower memory versions of `search`, `gather`, and `sketch`, as well as large-scale metagenome search (used for [petabyte-scale sequence search](https://www.biorxiv.org/content/10.1101/2022.11.02.514947v1)) and large-scale clustering. Read [the branchwater plugin docs](https://github.com/sourmash-bio/sourmash_plugin_branchwater/tree/main/doc/) for more information, and ask questions on [the sourmash issue tracker!](https://github.com/sourmash-bio/sourmash/issues) ### The `betterplot` plugin - improved plotting and visualization (Installable via pip as `sourmash_plugin_betterplot`.) The [`betterplot` plugin](https://github.com/sourmash-bio/sourmash_plugin_betterplot/) provides a variety of new plotting outputs for sourmash, including improved distance matrices, MDS plots, tSNE plots, upset plots, and Venn diagrams. It also supports cluster-cutting and extraction, as well as improved labeling and coloring by category. Read [the betterplot docs](https://github.com/sourmash-bio/sourmash_plugin_betterplot/) for more information, and ask questions on [the sourmash issue tracker!](https://github.com/sourmash-bio/sourmash/issues) ### The `directsketch` plugin - streaming download and sketch (Installable via conda and pip as `sourmash_plugin_directsketch`.) The [`directsketch` plugin](https://github.com/sourmash-bio/sourmash_plugin_directsketch/) provides two parallelized methods for streaming download and sketch from online sources. It provides two utilities: `gbsketch` for sketching NCBI Assembly Datasets, and `urlsketch` for sketching from generic URLs. Read [the directsketch docs](https://github.com/sourmash-bio/sourmash_plugin_directsketch/) for more information, and ask questions on [the sourmash issue tracker!](https://github.com/sourmash-bio/sourmash/issues)