New discoveries of Geometridae (Lepidoptera) from the extreme southwest of the Russian Far East – result of climate impact?

The increasing number of Barcode of Life Database (BOLD) records per species and genus leads to contradictory species assignments within Barcode Index Numbers (BINs), serving as identifiers for the BOLD ID engine. To examine these issues, we analyzed a dataset comprising original and curated BOLD records for the genus Tachina (Insecta: Tachinidae), based on a previous publication. This dataset included both published and private records. We were able to assess the performance of the BOLD engine’s species determination algorithm, Refined Single Linkage (RESL), and compare it to Assemble Species by Automatic Partitioning (ASAP). Additionally, we investigated the usage of BINs by the BOLD v4 ID engine. Our analysis confirmed that BOLD queries primarily rely on BINs for species identification, although some cases deviated from this pattern, resulting in species matches inconsistent with the assigned BIN species. ASAP was found to be superior to RESL due to RESL’s adherence to the concept of the DNA barcoding gap. Moreover, we found that taxonomic misassignments, inconsistencies in BIN formation, and missing metadata also contribute significantly to unreliable identifications. These problems appear to stem from both algorithmic limitations and deficiencies in submission and post-submission processes. Moreover, we noted that the default mode of the BOLD v4 ID engine integrates both private and published data, leading to public records based solely on COI-based identifications. However, this issue may now be mitigated, as the BOLD v5 ID engine default mode exclusively employs published data. To enhance BOLD’s reliability, we propose improvements to submission and post-submission processes. Without such amendments, the accumulation of contradictory species assignments within BINs will continue to rise and the reliability of specimen identification by BOLD will decrease.

South Africa is the third most biodiverse country in the world and insects represent a large part of its faunal diversity, as is seen globally. With more than 65,000 described animal species in South Africa, insects represent 44,088 species. While there are still a lot of species yet to be identified, progress may be hindered by the few insect taxonomists available in South Africa and subsequently, the time-consuming nature and costs of the methods used during species identification. DNA barcoding, on the other hand, has become a valuable tool for documenting biodiversity with the use of a small DNA fragment such as cytochrome oxidase subunit 1 (COI). This paper aims to assess South Africa's contribution to the Barcode of Life Database (BOLD) as well as highlight the regions that are under-represented on BOLD. From the 4,984,215 Insecta records on BOLD, South Africa contributed 56,392 insect records, with only 16.85% of that total identified to species level. The Gauteng Province had the most represented insect samples submitted to BOLD with 63.57% followed by Kwazulu-Natal (15.74%), and Mpumalanga (5.73%). However, the Free State, Limpopo, and the Northern Cape provinces are all under-represented on BOLD. This is evident asboth the Northern Cape and Limpopo provinces contain one or more biodiversity hotspots which in turn displays the high levels of biodiversity that could still be recorded on BOLD. Improving our understanding with regards to DNA barcoding data linked to geographical regions, as well as specific insect groups, can highlight the areas in need of more research.

Species identification techniques commonly utilized in Australian Forensic Science laboratories are gel immunodifussion antigen antibody reactions and hair comparison analysis. Both of these techniques have significant limitations and should be considered indicative opinion based tests. The Barcode of Life Initiative aims to sequence a section of DNA (~648 base pairs) for the Cytochrome Oxidase I mitochondrial gene (COI) in all living species on Earth, with the data generated being uploaded to the Barcode of Life Database (BOLD) which can then be used for species identification. The COI gene therefore offers forensics scientists an opportunity to use the marker to analyze unknown samples and compare sequences generated in BOLD. Once sequences from enough species are on the database, it is anticipated that routine identification of an unknown species may be possible. However, most forensic laboratories are not yet suited to this type of analysis and do not have the expertise to fully interpret the implications of matches and non matches involving a poorly sampled taxa (for example where there are cryptic species) and in providing the required opinion evidence. Currently, the use of BOLD is limited by the number of relevant species held in the database and the quality assurance and regulation of sequences that are there. In this paper, the COI methodology and BOLD are tested on a selection of introduced and Australian mammals in a forensic environment as the first step necessary in the implementation of this approach in the Australian context. Our data indicates that the COI methodology performs well on distinct species but needs further exploration when identifying more closely related species. It is evident from our study that changes will be required to implement DNA based wildlife forensics using the BOLD approach for forensic applications and recommendations are made for the future adoption of this technology into forensic laboratories.

DNA barcoding as a regulatory tool for seafood authentication in Canada

Metabarcoding and metagenomic approaches are becoming routine techniques in biodiversity assessment and ecological studies. The assignment of taxonomic information to sequences is challenging, as many reference libraries are lacking information on certain taxonomic groups and can contain erroneous sequences. Combining different reference databases is therefore a promising approach for maximizing taxonomic coverage and reliability of results. This tutorial shows how to use the “BOLD_NCBI_Merger” script to combine sequence data obtained from the National Center for Biotechnology Information (NCBI) GenBank and the Barcode of Life Database (BOLD) and prepare it for taxonomic assignment with the software MEGAN.

Trade in freshwater ornamental fish in South Africa is currently regulated by a ‘blacklist’ to prevent potentially invasive taxa from establishing in the country. Because its effective implementation requires accurate identification, the aim of the present study was to test whether DNA barcoding is a useful tool to identify freshwater fishes in the South African pet trade. A total of 351 aquarium fish specimens, representing 185 traded taxa, were sequenced for the mitochondrial COI barcoding marker in 2011 and 2012. Lake Malawi cichlids were treated as a single group due to a lack of resolution in their COI marker, resulting in a data set of 137 successfully sequenced taxa. The Barcode Of Life Database (BOLD) and GenBank were used for taxonomic assignment comparisons. The genetic identification matched the scientific name inferred from the trade name for 60 taxa (43.8%) using BOLD, and for 67 taxa (48.9%) using GenBank. A genetic ID could not be assigned in 47 (34.3%) cases using BOLD and in 37 cases (27%) using GenBank. Whereas DNA barcoding can be a useful tool to help identify imported freshwater fishes, it requires further development of publicly available databases to become a reliable means of identification.

In recent years, large‐scale DNA barcoding campaigns have generated an enormous amount of COI barcodes, which are usually stored in NCBI's GenBank and the official Barcode of Life database (BOLD). BOLD data are generally associated with more detailed and better curated meta‐data, because a great proportion is based on expert‐verified and vouchered material, accessible in public collections. In the course of the initiative German Barcode of Life data were generated for the reference library of 2,846 species of Coleoptera from 13,516 individuals. Confronted with the high effort associated with the identification, verification and data validation, a bioinformatic pipeline, “TaxCI” was developed that (1) identifies taxonomic inconsistencies in a given tree topology (optionally including a reference dataset), (2) discriminates between different cases of incongruence in order to identify contamination or misidentified specimens, (3) graphically marks those cases in the tree, which finally can be checked again and, if needed, corrected or removed from the dataset. For this, “TaxCI” may use DNA‐based species delimitations from other approaches (e.g. mPTP) or may perform implemented threshold‐based clustering. The data‐processing pipeline was tested on a newly generated set of barcodes, using the available BOLD records as a reference. A data revision based on the first run of the TaxCI tool resulted in the second TaxCI analysis in a taxonomic match ratio very similar to the one recorded from the reference set (92% vs. 94%). The revised dataset improved by nearly 20% through this procedure compared to the original, uncorrected one. Overall, the new processing pipeline for DNA barcode data allows for the rapid and easy identification of inconsistencies in large datasets, which can be dealt with before submitting them to public data repositories like BOLD or GenBank. Ultimately, this will increase the quality of submitted data and the speed of data submission, while primarily avoiding the deterioration of the accuracy of the data repositories due to ambiguously identified or contaminated specimens.

This study examines the genetic data coverage and availability in the Barcode of Life Database (BOLD), versions 2.5 and 3.0, and GenBank for the 88 invasive insects listed in the Global Invasive Species Database (http://www.issg.org). No data are recorded in either BOLD or GenBank for seven of those species. As a dedicated repository of curated barcode data BOLD is either missing data or contains inaccessible private data for 37 (42%) of the species while no data are available in GenBank for nine (8%) of the species. An evaluation of the Barcode Identification Number (BIN) scheme in BOLD ver. 3.0 was also evaluated and in 41% of cases the BIN contained more than one species. This essentially arose due to the 1% delimitation thresholds associated with the BINs and would result in misidentifications. Overall, more information is available from GenBank for the 88 invasive species listed on the Global Invasive Species Database, but quality checking is required to ensure that the data extracted from GenBank are of sufficient quality to make it useful. The implications of these results are discussed, with investment in parallel data silos suggested to be both costly and potentially an inefficient use of resources that may lead to loss of data if the means needed to maintain these databases become unavailable.

Metabarcoding and metagenomic approaches are becoming routine techniquesfor use in biodiversity assessment and in ecological studies. The assignment of taxonomic information to millions of sequences obtained via high-throughput sequencing is challenging, as many DNA reference libraries are lacking information on certain taxonomic groups and can contain erroneous sequences. Combining different reference databases is therefore a promising approach for maximising taxonomic coverage and reliability of results. The “BOLD_NCBI_Merger” bash script is introduced, which combines sequence data obtained from the National Centre for Biotechnology Information (NCBI) GenBank and the Barcode of Life Database (BOLD) and prepares it for taxonomic assignment with the software MEGAN.

With current trends in global warming, it has been suggested that spruce budworm outbreaks may spread to northern parts of the boreal forest. However, the major constraints for a northward expansion are the availability of suitable host trees and the insect winter survival capacity. This study aimed to determine the effect of larval feeding on balsam fir, white spruce and black spruce on various spruce budworm life history traits of both the parental and the progeny generations. Results indicated that the weight of the overwintering larval progeny and their winter survival were influenced by host tree species on which larvae of the parental generation fed. White spruce was the most suitable host for the spruce budworm, producing the heaviest pupae and the heaviest overwintering larvae while black spruce was the least suitable, producing the smallest pupae and the smallest overwintering progeny. Overwintering larvae produced by parents that fed on black spruce also suffered higher winter mortality than individuals coming from parents that fed on balsam fir or white spruce. With current trends in global warming, spruce budworm is expected to expand its range to northern boreal forests where black spruce is the dominant tree species. Such northern range expansion might not result in outbreaks if low offspring winter survival on black spruce persist.

Current trends in Northern Hemisphere and Central England temperatures are estimated using a variety of statistical signal extraction and filtering techniques and their extrapolations are compared with the pre dictions from coupled atmospheric-ocean general circulation models. Ear lier warming trend epochs are also analysed and compared with the current warming trend, suggesting that the long-run patterns of temperature trends should also be considered alongside the current emphasis on global warming.

Genetic diversity of polychaete annelids from the Saint Peter and Saint Paul Archipelago, Equatorial Atlantic, with description of a new species

Species extinctions in the tropics are accelerating, outpacing documentation efforts. Meanwhile, DNA barcoding is flourishing in the Global North, backed by extensive infrastructure, allowing non-taxonomic experts to identify species from nonlethal, minimally invasive, and environmental samples. However, hyper-diverse regions like Peru make up only 0.52% (n = 93,246) of the Barcode of Life Database (BOLD). To address this, we established three decentralized laboratories with low-cost, portable nanopore sequencers. From 2018–2023, we generated 1,858 barcodes in situ using six genetic markers for 1,097 vertebrates and 76 plants from existing and new biobanks. We present the first genetic barcodes for 30 mammal and 196 bird species from Peruvian specimens, increasing the number of Peruvian mammal and bird species in BOLD by 110% and 36.5% respectively. We also report the first records of the marsupial Marmosops ocellatus and the bat Sturnira lilium for Peru. This dataset represents an effort to go from fresh or museum-preserved samples to barcodes entirely in situ, avoiding the export of samples outside the country, and facilitating local capacity in molecular biodiversity research.

DNA barcode sequences were developed from 557 mesopelagic and upper bathypelagic teleost specimens collected in waters off Atlantic Canada. Confident morphological identifications were available for 366 specimens, of 118 species and 93 genera, which yielded 328 haplotypes. Five of the species were novel to the Barcode of Life Database (BOLD). Most of the 118 species conformed to expectations of monophyly and the presence of a “barcode gap”, though some known weaknesses in existing taxonomy were confirmed and a deficiency in published keys was revealed. Of the specimens for which no firm morphological identification was available, 156 were successfully identified to species, and a further 11 to genus, using their barcode sequences and a combination of distance- and character-based methods. The remaining 24 specimens were from species for which no reference barcode is yet available or else ones confused by apparent misidentification of publicly available sequences in BOLD. Addition of the new sequences to those previously in BOLD contributed support to recent taxonomic revisions of Chiasmodon and Poromitra, while it also revealed 18 cases of potential cryptic speciation. Most of the latter appear to result from genetic divergence among populations in different ocean basins, while the general lack of strong horizontal environmental gradients within the deep sea has allowed morphology to be conserved. Other examples of divergence appear to distinguish individuals living under the sub-tropical gyre of the North Atlantic from those under that ocean’s sub-polar gyre. In contrast, the available sequences for two myctophid species, Benthosema glaciale and Notoscopelus elongatus, showed genetic structuring on finer geographic scales. The observed structure was not consistent with recent suggestions that “resident” populations of myctophids can maintain allopatry despite the mixing of ocean waters. Rather, it indicates that the very rapid speciation characteristic of the Myctophidae is both on-going and detectable using barcodes.

DNA barcoding was proposed in 2003, the Consortium for the Barcode of Life was established in 2004, and the movement has since attracted more than $80 million funding. Here we investigate how many species of multicellular animals have been barcoded. We compare the numbers in a public database (GenBank as of January 2012) with those in the Barcode of Life Database (BOLD) and find that GenBank contains COI (cytochrome c oxidase subunit 1) sequences for ca. 60 000 species while BOLD reports barcodes for ca. 150 000 species. The discrepancy is likely due to a large amount of unpublished data in BOLD. Overall, the species coverage remains sparse, growth rates are low, and the barcode accumulation curve for Metazoa is linear with only 4788 species having been added in 2011. In addition, the vast majority of species in the public database (73%) were barcoded by projects that are unlikely to be related to the DNA barcoding movement. Particularly surprising was the large number of DNA barcodes in GenBank that were not identified to species (Jan 2012: 74%), with insect barcodes often being identified only to order. Of these several hundred thousand have since been suppressed by NCBI because they did not satisfy the iBOL/GenBank early release agreement. Species coverage is considerably better for target taxa of DNA barcoding campaigns (e.g. birds, fishes, Lepidoptera), although it also falls short of published campaign targets. © The Willi Hennig Society 2012.