DNA barcodes are not only used by researchers, but also by decision-makers (e.g. to control food fraud or illegal species commercialization). The big-scale demand of both online services and information to identify species contrasts with the limited ways to automatize either species identification or assessment of barcode quality per species, directly from the web interface.
BOLD system is the main database of DNA barcode worldwide. This database has been stepply growing through time since its release (Ratnasingham and Hebert 2007) and its accessibility is pivotal for projects focused on DNA barcodes. Nowdays APIs, to some extant, are offering access to well-know databases such as FishBase, WoRMS or BOLD. Despite BOLD's API mostly involves only public data, this leverages its data retrieving for wider purposes. The API's applicability, however, seems to be wholly held up by its own needs of having either standalone softwares or functions which could wrap up blocks of information. The main objective of these functions (i.e. BOLD-mineR's functions) is justly circumscribe the BOLD's API performance with R-based scripts to get insights about DNA barcodes by using public information.
You can install the development version from GitHub with:
# library(devtools)
devtools::install_github("Ulises-Rosas/boldminer")library(boldminer)This function lets us to mine associated metadata from any specimen according to following arguments:
taxon(e.g.Aves|Elasmobranchii).ids(e.g.ANGBF12704-15).bin(e.g.BOLD:AAA4689).container(e.g.FIPP).institution(e.g.Smithsonian Institution).researchers(including identifiers and collectors).geo(e.g.Peru).
If we want to get information, for instance, from all specimens of elasmobranchs distributed in Peru and stored in BOLD, we can use the following line:
specimendata <- boldminer::SpecimenData(taxon = "Elasmobranchii", geo = "Peru")Then,we use tibble package for just assessing specimendata dimension:
tibble::as_tibble(specimendata)
#> # A tibble: 99 x 68
#> processid sampleid recordID catalognum fieldnum institution_sto…
#> <fct> <fct> <int> <fct> <fct> <fct>
#> 1 ANGBF109… KJ146022 5651960 "" KJ146022 Mined from GenB…
#> 2 ANGBF109… KJ146024 5651962 "" KJ146024 Mined from GenB…
#> 3 ANGBF109… KJ146025 5651963 "" KJ146025 Mined from GenB…
#> 4 ANGBF110… KJ146027 5652062 "" KJ146027 Mined from GenB…
#> 5 ANGBF110… KJ146021 5652090 "" KJ146021 Mined from GenB…
#> 6 ANGBF110… KJ146043 5652112 "" KJ146043 Mined from GenB…
#> 7 ANGBF110… KJ146042 5652113 "" KJ146042 Mined from GenB…
#> 8 ANGBF116… KJ146045 5652680 "" KJ146045 Mined from GenB…
#> 9 ANGBF116… KJ146041 5652681 "" KJ146041 Mined from GenB…
#> 10 ANGBF116… KJ146040 5652682 "" KJ146040 Mined from GenB…
#> # … with 89 more rows, and 62 more variables: collection_code <lgl>,
#> # bin_uri <fct>, phylum_taxID <int>, phylum_name <fct>,
#> # class_taxID <int>, class_name <fct>, order_taxID <int>,
#> # order_name <fct>, family_taxID <int>, family_name <fct>,
#> # subfamily_taxID <lgl>, subfamily_name <lgl>, genus_taxID <int>,
#> # genus_name <fct>, species_taxID <int>, species_name <fct>,
#> # subspecies_taxID <lgl>, subspecies_name <lgl>,
#> # identification_provided_by <fct>, identification_method <fct>,
#> # identification_reference <fct>, tax_note <lgl>, voucher_status <fct>,
#> # tissue_type <fct>, collection_event_id <lgl>, collectors <fct>,
#> # collectiondate_start <lgl>, collectiondate_end <lgl>,
#> # collectiontime <fct>, collection_note <lgl>, site_code <lgl>,
#> # sampling_protocol <fct>, lifestage <fct>, sex <lgl>,
#> # reproduction <fct>, habitat <fct>, associated_specimens <lgl>,
#> # associated_taxa <lgl>, extrainfo <fct>, notes <fct>, lat <dbl>,
#> # lon <dbl>, coord_source <lgl>, coord_accuracy <lgl>, elev <lgl>,
#> # depth <int>, elev_accuracy <lgl>, depth_accuracy <lgl>, country <fct>,
#> # province_state <fct>, region <fct>, sector <fct>, exactsite <fct>,
#> # image_ids <lgl>, image_urls <lgl>, media_descriptors <lgl>,
#> # captions <lgl>, copyright_holders <lgl>, copyright_years <lgl>,
#> # copyright_licenses <lgl>, copyright_institutions <lgl>,
#> # photographers <lgl>Given its dimension, we can summarize our specimendata data frame with the sumSData() utility:
boldminer::sumSData(df = specimendata, cols = c("species_name", "country"))
#> species_name country n
#> 1: Peru 58
#> 2: Isurus oxyrinchus Peru 14
#> 3: Prionace glauca Peru 9
#> 4: Alopias pelagicus Peru 5
#> 5: Lamna nasus Peru 3
#> 6: Galeorhinus galeus Peru 2
#> 7: Carcharhinus brachyurus Peru 1
#> 8: Carcharhinus obscurus Peru 1
#> 9: Sphyrna zygaena Peru 1
#> 10: Potamotrygon falkneri Peru 1
#> 11: Potamotrygon sp. a Peru 1
#> 12: Potamotrygon sp. Peru 1
#> 13: Aculeola nigra Peru 1
#> 14: Urotrygon chilensis Peru 1Where n column shows up unique counts after grouping data frame by values from cols argument. You can also plot geographical information, when available, by using leafletPlot():
boldminer::leafletPlot(specimendata)
If only sequences are desired, the argument seq = "only" should be stated. You can also combine metadata with sequences by using seq = "combined"
seqs <- boldminer::SpecimenData(taxon = "Elasmobranchii", geo = "Peru", seq = "only")
seqs
#> 99 DNA sequences in binary format stored in a list.
#>
#> Mean sequence length: 641.899
#> Shortest sequence: 226
#> Longest sequence: 712
#>
#> Labels:
#> ANGBF10913-15|Alopias pelagicus|COI-5P|KJ146022
#> ANGBF10915-15|Alopias pelagicus|COI-5P|KJ146024
#> ANGBF10916-15|Alopias pelagicus|COI-5P|KJ146025
#> ANGBF11015-15|Carcharhinus brachyurus|COI-5P|KJ146027
#> ANGBF11043-15|Carcharhinus obscurus|COI-5P|KJ146021
#> ANGBF11065-15|Prionace glauca|COI-5P|KJ146043
#> ...
#>
#> Base composition:
#> a c g t
#> 0.258 0.254 0.166 0.323
#> (Total: 63.55 kb)Above sequences can be also exported into a file by using write.dna() function from the ape package, which, in turn, the boldminer package depends:
# library(ape)
ape::write.dna(x = seqs, file = 'sequences.txt', format = 'fasta', nbcol = 1, colw = 90)This function finds best matches between a query sequence and a database of BOLD by using BLASTn-based algorithms. Arguments of this function are query and db. The first one are query sequences and the second one are one of avilable databases in BOLD:
COX1COX1_SPECIESCOX1_SPECIES_PUBLICCOX1_L640bp
This script also take account for those sequences which are not, by mistake, at the right sense and sends them to perform a BLAST search through its API
This function starts with a fasta-formated file. In order to test this script, we can take sample file within the package:
fasta_file <- system.file("sequences.fa", package = "boldminer") Then, we can run the following line get its identification:
id_out <- boldminer::ID_engine(query = fasta_file, db = "COX1_SPECIES")First five rows for each id_out item can be taken by using lookID() function:
boldminer::lookID(id_out)
#> Sample ID taxonomicidentification similarity
#> 1 seq1 PHANT458-08 Alopias pelagicus 1
#> 2 seq1 IRREK872-08 Alopias pelagicus 1
#> 3 seq1 IRREK873-08 Alopias pelagicus 1
#> 4 seq1 IRREK874-08 Alopias pelagicus 1
#> 5 seq1 IRREK876-08 Alopias pelagicus 1
#> 6 seq2 ANGBF10914-15 Alopias pelagicus 0.9986
#> 7 seq2 ESHKB029-07 Alopias pelagicus 0.9985
#> 8 seq2 ANGBF10913-15 Alopias pelagicus 0.9971
#> 9 seq2 PHANT458-08 Alopias pelagicus 0.9969
#> 10 seq2 IRREK873-08 Alopias pelagicus 0.9969
#> 11 seq3 ANGBF12626-15 Alopias pelagicus 1
#> 12 seq3 ANGBF12623-15 Alopias pelagicus 1
#> 13 seq3 ANGBF11723-15 Alopias pelagicus 1
#> 14 seq3 ANGBF10915-15 Alopias pelagicus 1
#> 15 seq3 ESHKB036-07 Alopias pelagicus 1This function adds an audition step (Oliveira et al. 2016) to each selected specimen by ID_engine() (see above), given a certain threshold. This function, in turn, uses another function called AuditionBarcodes() (see below). This prior function is coupled with auditOnID() and can validate species names by taking accepted names from Worms database.
As seen with ID_engine(), this function starts with a fasta-formated file. We can take the same sample file we used: fasta_file. Then, we can identify those sequences under audiOnID() functionality by using a threshold = 0.99 (i.e. 99% of similarity):
aoID_out <- boldminer::auditOnID(seqs = fasta_file, threshold = 0.99)
#> |*****************************************************************| 100%aoID_out
#> Samples Match Species Grades Observations
#> 1 seq1 Unique Alopias pelagicus A There were 49 matches. External congruence.
#> 2 seq2 Unique Alopias pelagicus A There were 39 matches. External congruence.
#> 3 seq3 Unique Alopias pelagicus A There were 94 matches. External congruence.We can also skip audition step by adding just_ID = TRUE within arguments.
Despite AuditionBarcodes() function is coupled with auditOnID() function, it can also work with just a list of names. Furthermore, there is an argument which enables to chose if sequences from GenBank are considered. It is pending, however, assess whether these sequences used to assess barcode's quality come from either a published article or direct submission:
species <- c( "Caretta caretta", "Bathygobius lineatus",
"Albula esuncula", "Vibilia armata",
"Alepisaurus ferox", "Diodon hystrix",
"Vesicomya galatheae", "Caranx ruber")
audit_out <- boldminer::AuditionBarcodes(species, exclude_ncbi = T, validate_name = T)
#> Auditing for:
#>
#> Caretta caretta
#> Bathygobius lineatus
#> Eretmochelys imbricata
#> Vibilia armata
#> Alepisaurus ferox
#> Diodon hystrix
#> Vesicomya galatheae
#> Caranx ruberaudit_out
#> Species Grades Observations BIN_structure
#> 1 Caretta caretta A Matched BIN with external congruence 'BOLD:AAB8364':{'Caretta caretta':11}
#> 2 Bathygobius lineatus B Matched BIN with internal congruence only 'BOLD:AAF0181':{'Bathygobius lineatus':4}
#> 3 Albula esuncula C Splitted BIN 'BOLD:AAF1162':{'Albula esuncula':3}, 'BOLD:AAA3538':{'Albula esuncula':1}
#> 4 Vibilia armata D Insufficient data. Institution storing: 2. Specimen records: 20 <NA>
#> 5 Alepisaurus ferox E** Mixtured BIN 'BOLD:AAC5235':{'Alepisaurus ferox':8}, 'BOLD:AAC5236':{'Alepisaurus brevirostris':3,'Alepisaurus ferox':3}
#> 6 Diodon hystrix E* Merged BIN 'BOLD:AAB0446':{'Diodon hystrix':17,'Diodon eydouxii': 1}
#> 7 Vesicomya galatheae F Barcodes mined from GenBank or unvouchered. <NA>
#> 8 Caranx ruber NA No specimen data available <NA>Please notice that grades are obtained with accepted names of species according to WoRMS database Rest service by using its taxamatch algorithm. Hence, since currently accepted names within species vector has not been figured out, unevenness between the column BIN_structure and species could pop up.