ABSTRACT This study presents DNA barcoding data for Alpiscorpius dinaricus (Di Caporiacco) and Euscorpius hadzii Caporiacco. Barcode sequences were compared with publicly available reference data to support species identification, together with the evaluation of diagnostic morphological characters. Alpiscorpius dinaricus is recorded from Kosovo for the first time, representing a new national record and contributing to the knowledge of euscorpiid diversity in the region. Diagnostic illustrations of A. dinaricus are provided to facilitate reliable identification and to support future faunistic, taxonomic, and biogeographic studies. Euscorpius hadzii, previously known only from Prizren district, is now reported also from Bjeshkët e Nemuna Mountains (Western Kosovo).

DNA barcoding markers: A comprehensive review and taxonomic classification across species.

Assessing temporal and spatial variation in the diet of the Common Kestrel (Falco tinnunculus) using web-sourced photography

Species delimitation has been facilitated by the utilization of molecular markers, which, within the framework of integrative taxonomy, has contributed to a more profound comprehension of biodiversity. The analysis of DNA sequences of sand flies has yielded significant progress in species identification, sex association, and the description of species complexes, but has been overlooked concerning Brumptomyia França & Parrot, 1921 genus. Here, we assessed the molecular taxonomy of some Brumptomyia species, highlighting the challenges encountered in delimiting these taxa by different methods. We have sequenced and analyzed the DNA barcoding fragment of the cytochrome c oxidase subunit I (COI) gene, in addition to a nuclear DNA marker of the paralytic (PARA) gene. The final dataset encompasses 11 Brumptomyia species, 149 COI sequences, and 30 PARA fragments. A set of different discovery and validation species-delimitation approaches were performed, including single-locus heuristic ones, and an approach based on the multispecies coalescent model (MSC). Interspecific pairwise genetic distances were found to be lower than intraspecific distances. In a similar manner, the species pairs B. avellari/B. brumpti, B. cavicola/B. troglodytes, and B. nitzulescui/B. mangabeirai were merged into a single genetic cluster for each pair. Consequently, only five Brumptomyia taxa could be unambiguously identified using the standard DNA barcoding approach. However, the validation analysis under the MSC indicates that nearly all nominal species represent independent taxa, except for B. avellari and B. brumpti. These two latter, although closely-related, are well recognized by their morphology, and possibly more molecular markers, individuals, and populations are needed to understand their taxonomic status and evolutionary relationship. The species delimitation of Brumptomyia remains challenging, and further genomic assessments are necessary to advance our understanding of its diversity.

Spatiotemporal patterns of bird strikes at civil airports across China inferred from DNA barcoding (2012–2024)

Simple, rapid and cost-effective DNA extraction techniques for detection of economically important fruit flies in India.

Lagerstroemia minuticarpa Debb. ex P. C. Kanjilal is a deciduous tree listed as a nationally protected wild plant (level II) in China. The complete chloroplast genome of L. minuticarpa was sequenced using the DNBSEQ-T7 platform. The genome is 152,183 bp in length and exhibits a typical quadripartite structure, consisting of a large single-copy (LSC) region of 84,009 bp, a small single-copy (SSC) region of 16,924 bp, and a pair of inverted repeats (IRs) of 25,625 bp each. It contains 133 genes in total, including 85 protein-coding genes, 37 transfer RNA genes, and 8 ribosomal RNA genes. The overall GC content of the genome is 37.31%. Phylogenetic analysis based on 39 chloroplast genomes revealed that L. minuticarpa forms an independent lineage nested within the genus Lagerstroemia. Additionally, pairwise alignment identified ten highly variable chloroplast loci (including both coding and intergenic regions) with elevated parsimony-informative sites, which hold promise as specific DNA barcodes for species identification in Lagerstroemia. These data enrich the genetic knowledge of Lagerstroemia species in the Lythraceae family and provide a basis for further research in molecular identification, genetic breeding, and other related fields.

The highlands of the eastern Democratic Republic of the Congo (DRC) are home to critically endangered eastern gorillas (Gorilla beringei). Concerns have been raised that the increased temperatures and extreme weather conditions associated with climate change will lead to an increase in the abundance and distribution of Culicoides-borne diseases. Here, we utilized an integrated morphological and molecular approach to identify Culicoides species in a small but significant collection of Culicoides captured from highland eastern gorilla habitat and surrounding areas and updated the Culicoides spp. reported from the highlands of the eastern DRC. A review of the literature related to Culicoides collections in the DRC was conducted in French and English. Recent worldwide checklists were consulted to rectify synonyms and other discrepancies found in the literature for the region. Fresh Culicoides specimens were collected, wings slide-mounted and remaining carcasses subjected to DNA extraction. A total of 82 Culicoides specimens were collected. From these, 75 high-quality DNA barcodes (658-bp of the mtDNA COI gene) were obtained, belonging to 14 distinct taxa, 11 of which were new records for the DRC, including C. bolitinos Meiswinkel, 1989, C. hortenis Khamala & Kettle, 1971, C. citroneus Carter, Ingram & Macfie, 1920, and C. radiomaculatus Khamala & Kettle, 1971, and seven species new to science (C. sp. nr. citroneus, C. sp. nr. glabripennis 1, C. sp. nr. glabripennis 2, C. sp. nr. kibatiensis 1, C. sp. nr. kibatiensis 2, C. sp. nr. neavei 1 and C. sp. nr. neavei 2), increasing the known Culicoides fauna of the DRC from 20 to 31. The presence of C. imicola Kieffer, 1913, C. enderleini Cornet & Brunhes, 1994 and C. neavei Austin, 1912, was confirmed. The potential health impact of the association of known Culicoides pathogen vectors with endangered gorillas is discussed.

DNA barcoding for the identification of swim bladders: An approach to international trade monitoring

DNA barcoding of ants from India: Insights into cryptic diversity, species boundaries, and taxonomic ambiguities

Single-locus species-delimitation methods using DNA barcoding data have made an important contribution to large scale biodiversity inventories and integrative taxonomy, allowing rapid and repeatable estimates of alpha diversity from huge numbers of individuals. A range of distance-based and tree-based methods now exist to perform these analyses, including ABGD, ASAP, GMYC, and mPTP. However, the outputs of such software are idiosyncratic and difficult to integrate and compare. The R package delimtools offers a platform capable of: (1) pre-processing, cleaning and formatting data; (2) collapsing and summarising haplotype information; (3) executing, parsing and merging program outputs and species partitions; and (4) tabulating results, and visualizing on a phylogenetic tree. The open-source delimtools package opens up new avenues to scale up and standardize species delimitation analyses, and is available on the CRAN (https://cran.r-project.org/package=delimtools) and GitHub (https://github.com/legalLab/delimtools/) repositories.

Almonds (Prunus dulcis) (Mill.) are an economically important crop in California, with an estimated value of $8.6 billion dollars annually. The aim of this study was to identify the hemipteran pest and beneficial species in conventionally managed almond orchards when the developing almond crop was vulnerable to feeding damage. Insects were collected in 9 orchards by multiple methods and then Hemiptera were sorted to morphospecies. The DNA barcode of the mitochondrial DNA COI gene was sequenced for each specimen. For each species, the haplotype diversity was determined. A high diversity of Hemiptera was collected, including 26 species from 12 families. Pest species included Halyomorpha halys Stål (Hemiptera: Pentatomidae), Chinavia hilaris (Say) (Hemiptera: Pentatomidae), Lygus hesperus Knight, and Leptoglossus zonatus (Dallas) (Hemiptera: Coreidae); beneficials included Brochymena quadripustulata (Fabricius) (Hemiptera: Pentatomidae), Orius spp. Wolff (Hemiptera: Anthocoridae), Nabis spp. Latreille (Hemiptera: Nabidae) and Geocoris atricolor Montandon (Hemiptera: Geocoridae). Several orchards had high species diversity (13 species), others had only one species recovered. Haplotype diversity was low in most pest species except for L. hesperus (0.6) and was high in 2 beneficial groups, Nabis spp. and Orius tristicolor (White) (0.6 to 0.76). Hemipteran pests were detected through the time when developing almonds were susceptible to feeding damage, while beneficial Hemiptera were more abundant later. Correct identification of insect species is an integral component to Integrated Pest Management (IPM) to trace the origin of invasive species, uncover cryptic species, and to develop a relationship between species and crop damage.

Four new species of the genus Macrobiotus are described from Asia and Africa: Macrobiotus witalinskii sp. nov., M. dalaticus sp. nov., M. surmaczi sp. nov., and M. hoianicus sp. nov. Species delimitation is based on detailed morphological and morphometric analyses using phase-contrast light microscopy and scanning electron microscopy, complemented by molecular data from four commonly used genetic markers. Phylogenetic analyses consistently place all four species within Macrobiotus clade C, each forming a distinct, well-supported monophyletic lineage. The newly described taxa expand the known morphological and ecological diversity of clade C and show that egg and body coloration characters previously considered diagnostic for this lineage are not evolutionarily conserved. These results further emphasize the value of integrative taxonomy for documenting hidden diversity and for supporting future biodiversity assessments, including large-scale DNA barcoding and metabarcoding surveys of tardigrades.

This study investigates the seasonal ecology, morphology, and invasion history of the aphids Cinara atlantica and C. watsoni (Hemiptera: Aphididae: Lachninae) in South Korea and assesses their potential ecological impacts. Historically, C. atlantica was recorded only on non-native pines, but we provide the first evidence of host expansion to the native pine Pinus densiflora. C. watsoni is recorded for the first time in South Korea on imported pines, indicating a potential risk of spread to native hosts. Field surveys conducted between 2020 and 2025, including monthly monitoring, showed that both species reproduce anholocyclically, with colony densities peaking in June and October-November. Honeydew accumulation promoted sooty mold development, and heavy infestations caused needle yellowing. Morphological redescriptions, DNA barcoding, and haplotype analyses confirmed species identities and revealed minimal genetic divergence between South Korean and Nearctic populations. Shared haplotypes with North American material suggest a recent introduction possibly mediated by international plant trade, although the precise source remains uncertain. These findings highlight the high invasion potential of Cinara species and emphasize the need for strengthened phytosanitary measures and continuous monitoring of conifer-associated aphids in East Asia.

Bacteriophage (phage)-based vectors offer considerable promise as tools for tuning the microbiome with molecular and genetic precision. However, standardized methods to rigorously characterize phage vectors remain lacking. Here, we present an optimized digital droplet PCR (ddPCR)-based assay for quantifying both the purity and potency of phage vector preparations. We utilized central composite design to develop a ddPCR assay capable of quantifying the number of phage vector capsids packed with the phage vector genome or packed with the transgenic DNA of interest. This assay targets 2 unique DNA barcodes, designed to be biologically inert and maximally orthogonal to existing DNA sequences. Through stringent optimization, we were able to achieve assay conditions that enable a dynamic range of nearly 3 orders of magnitude and correct for systemic error in the assay. We then show that biological activity assays consistently underestimate transgene-packed vector titers, leading to overestimation of true transduction efficiency, particularly when contamination by genome-packed vectors is high. We further demonstrate how this approach facilitates optimization of vector production conditions and substantially improves the precision and reproducibility of phage vector transduction. Compared to assays of biological activity, this optimized ddPCR assay has improved accuracy and, through design of experiments optimization, high precision (CVs = 5.5 ± 1.3% and 4.5 ± 1.0% for the genome and transgene barcodes, respectively). This assay can be broadly adopted to characterize and quality control vector preparations for various applications.

Nongmangkha is one of the several medicinal plants traditionally used by the Meitei community of Manipur, India, for treating respiratory and infectious diseases. However, taxonomic confusion exists among three commonly used species- Phlogacanthus thyrsiformis(Roxb. ex Hardw.) Mabb, Phlogacanthus pubinerviusT. Anderson, and Justicia adhatodaL.-limiting their safe and effective utilization. Three chloroplast gene- rbcl, rpoC1 and trnH-psbA were used as DNA barcodes to identify the plants then, the sequences obtained were used for secondary structure prediction in RNAfold web server to discriminate among the three plants. The HPTLC revealed chemical fingerprints of the studied species which helped in taxonomic authentication of plants, while GC-MS and FTIR analysis of the extracts showed presence of several bioactive compounds and functional groups of the compounds respectively. Elemental analysis using XRF detected 19 elements in P. pubinervius indicating higher chemical diversity, while J. adhatoda and P. thyrsiformis contained 15 elements each. The plant extracts examined demonstrated significant antimicrobial activity against Staphylococcus epidermidis, Pseudomonas aeruginosa, Epidermophyton floccosum, and Fusarium oxysporum. To further explore their potential, molecular docking studies were carried out using the key bioactive compounds, which revealed promising interactions with Metallo-β-lactamase-suggesting their potential as effective antimicrobial agents. To the best of our knowledge, this is the first report to provide comparative insights into the three studied samples at anatomical, chemical, and molecular levels, paving the way toward accurate identification.

DNA barcode has become an important tool for identifying species, accelerating biodiversity assessments. However, most species still lack barcode data, especially in tropical biomes. We used a comprehensive DNA barcode library for South American tree species and identified geographic and phylogenetic gaps in DNA sequences. We compiled distribution data for 11,525 tree species across 3,283 sites in four ecoregions: Amazonia, Caatinga, Cerrado, and Mata Atlantica, and searched GenBank for the main plant DNA barcodes: ITS2, matk, rbcL and trnH-psbA. To assess phylogenetic diversity gaps, we used a dated molecular phylogeny of 1,100 angiosperm genera found in tropical South America. Only around 25% of species have sequences available in GenBank for the selected DNA markers. Most species lacking sequences occur in the Mata Atlantica and Amazonia ecoregions. Moreover, missing species on GenBank are not randomly distributed across the angiosperm phylogeny, leading to large phylogenetic gaps. Calculating phylogenetic endemism, we identified new priority sites for future fieldwork, highlighting the importance of biodiversity metrics beyond species richness. This work revealed major geographic and phylogenetic gaps in DNA barcode data for tropical tree species in South America, helping to guide targeted sampling and improve conservation planning based on biodiversity and evolutionary analyses.

West Nile virus (WNV) and eastern equine encephalitis virus (EEEV) are typically maintained via enzootic cycles between birds and mosquitoes (Diptera: Culicidae). In North America, the blood hosts of mosquitoes involved in pathogen circulation are poorly described for many regions, limiting our ability to predict disease outbreaks. Our objectives were to (i) identify local (Eastern Ontario, Canada) bloodmeal hosts fed on by mosquitoes using DNA barcoding and (ii) investigate if the bloodmeal hosts identified were representative of the entire host community using species accumulation curves and Chao richness estimates. We successfully identified 313 bloodmeal hosts from 292 mosquitoes belonging to the species Aedes cinereus, Aedes vexans, Coquillettidia perturbans, Culex pipiens/restuans, Ochlerotatus stimulans and Ochlerotatus trivittatus, with 257 and 56 bloodmeal hosts identified at the species and genus levels, respectively. Concurrent with the literature, Culex species almost exclusively fed on birds, C. perturbans fed on a mixture of birds and mammals, and the remaining mosquito species fed almost exclusively on mammals. Eastern cottontail rabbits (Sylvilagus floridanus) were the most common mammal species fed on, driven by A. vexans and O. trivittatus. With respect to WNV and EEEV enzootic cycles, Culex species, the main vectors for WNV transmission, were observed to forage on American robins (Turdus migratorius), an amplification host for WNV, highlighting a pathway by which WNV could be maintained in our study region. A. vexans, a potential bridge vector, was also observed to forage on a relatively high frequency of humans (Homo sapiens), suggesting a pathway by which humans could be exposed to WNV. C. perturbans, a potential bridge vector for EEEV transmission, was also observed to forage on both an American Robin (also an amplifying host for EEEV) as well as on humans in a relatively high frequency, illustrating a potential pathway by which EEEV exposure in humans could occur. Our species accumulation curves and Chao estimates of richness with respect to bloodmeal hosts, indicated that the bloodmeal host community was not completely characterized. Together, our results contribute to a better understanding of mosquito bloodmeal hosts in eastern North America and suggest a larger or more directed sampling effort would be needed to characterize the entire bloodmeal host community in our study region.

French Guiana is home to one of the highest mosquito diversities in the world, with currently 242 species recorded, nearly half of them belonging to the genus Culex. These mosquitoes include vectors of human parasites and viruses, some of which are of significant public health concern. Understanding the host preferences of female Culex mosquitoes is essential but knowledge is still limited. The Zoo of French Guiana was chosen as an experimental site for its constant availability of diverse captive animals and a potential sentinel role for monitoring infectious diseases. Between February 2018 and January 2019, 7892 adult mosquitoes were collected from 42 potential resting sites using manual aspiration. Blood-fed Culex accounted for 81% of the total number of blood-fed mosquitoes. Of these, 510 Culex female specimens were identified to species level by DNA barcoding, and their blood meals and parasites were analyzed using next-generation sequencing (NGS) technology. In total, 27 Culex species were identified at the zoo, and their blood meals revealed a broad range of vertebrate hosts, with relatively few being human. Culex eastor and Culex vaxus were the two most common species, feeding mainly on Tapirus terrestris and Iguana iguana, respectively. In addition, 60% of the mosquito species were infected with blood parasites, either known or novel (haemosporidian and kinetoplastid parasites). This study provides baseline insights into the diversity, the host associations, and the parasite communities of Culex mosquitoes in French Guiana. The zoo appears to be a valuable sentinel site, with mosquitoes feeding on a wide range of vertebrates, providing a foundation for future monitoring and research on mosquito-host-pathogen interactions.

Plants of the genus Bischofia (Phyllanthaceae) possess significant ornamental and cultural value and are widely used in urban landscaping. However, their morphological similarities make accurate species identification challenging. Although chloroplast genome sequences have become a crucial source of phylogenetic information for plant classification, the chloroplast genomes of Bischofia had not been deciphered within a comparative framework. In this study, we described key leaf anatomical traits and sequenced, assembled, and annotated the complete chloroplast genomes of Bischofia javanica and B. polycarpa to clarify their taxonomic boundaries. Anatomical investigation revealed significant divergence in leaf thickness, palisade mesophyll structure, and vascular traits. Both assembled chloroplast genomes displayed a typical circular quadripartite structure, with lengths of 162,495bp and 162,228bp, respectively. The B. javanica chloroplast genome contains 83 protein-coding genes, 8 rRNA genes, and 37 tRNA genes, while B. polycarpa contains 84 protein-coding genes, 8 rRNA genes, and 38 tRNA genes. Notably, the rpoC2 gene was present in B. polycarpa but absent in B. javanica-a condition validated by PCR using multiple individuals and primer sets and unique within Phyllanthaceae. Codon usage was biased toward A/U endings. Genome comparison revealed high nucleotide diversity in rpl22, ycf1, matK, clpP, and ndhF, suggesting their potential as DNA barcodes. Phylogenetic analysis confirmed a close relationship between the two Bischofia species and placed Bischofia, Baccaurea, and Antidesma in a distinct clade within Phyllanthaceae. This study provides the first integrated anatomical and chloroplast genomic resources for Bischofia. The complete chloroplast genomes, along with identified highly variable regions and the unique absence of rpoC2 in B. javanica, offer valuable tools for species identification, phylogenetic reconstruction, and understanding genome evolution in Phyllanthaceae.