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Despite the effectiveness of DNA metabarcoding for gaining insights into biodiversity and environmental species composition, a centralised management and storage option including easy accessibility of already-published data is lacking. Since most data are published as supplementary material or in private repositories, DNA metabarcoding has a huge untapped potential to be used for analyses across multiple taxa, sample locations or multiple research projects. We developed a platform to register, manage and identify amplicon sequence variants (ASVs) or zero-radius OTUs (ZOTUs), respectively, against several barcode reference datasets. Moreover, ASV tables can be uploaded, managed, versioned and published with DOIs thus contributing to the full research Data Life Cycle.

The genus Anacharis Dalman, 1823 comprises parasitoid wasps that target early instars of brown lacewing larvae (Neuroptera: Hemerobiidae). So far, five species were recognised from the Western Palaearctic region, of which four are reported from Northwestern Europe. In this study, we address the Northwestern European species diversity of the genus with an extended integrative taxonomy toolkit. A total of 700 specimens were examined for their external morphology, including the relevant type specimens. For 354 specimens, we obtained CO1 barcode sequences and applied three molecular species delimitation methods. All DNA barcode data are made publicly available via the German Barcode of Life (GBOL) and Barcode of Life Data system (BOLD) database. In addition, we examined images of Wing Interference Patterns (WIPs), examined the male genitalia and performed multivariate morphometric analyses. The analyses revealed two clusters which we describe as the immunis and eucharioides species groups based on differences in DNA barcode, external morphology, WIPs and size of the male genitalia. Furthermore, we complement the diagnosis of the genus Anacharis and describe three new species, Anacharis martinae Vogel, Forshage & Peters, sp. nov., Anacharis maxima Vogel, Forshage & Peters, sp. nov. and Anacharis minima Vogel, Forshage & Peters, sp. nov. Finally, we synonymise A. fergussoni Mata-Casanova & Pujade-Villar, 2018, syn. nov. with A. eucharioides (Dalman, 1818), and we reinstate A. ensifer Walker, 1835, stat. rev., A. typica Walker, 1835, stat. rev. and A. petiolata Zetterstedt, 1838, stat. rev. as valid species. In total, we recognise nine Northwestern European species to which we provide an identification key. The species of Anacharis are morphologically very variable. Morphometric analyses alone did not provide information sufficient to delimit species, neither did analyses of WIPs and male genitalia, with few notable exceptions. Analyses of molecular sequence data proved crucially helpful to reliably delimit species and to find morphological diagnostic characters in a reverse taxonomy approach. For delimiting species groups, all included analyses proved helpful, and we show that exploring an extended integrative taxonomy toolkit can be beneficial for a comprehensive characterisation of species. We acknowledge that a complete overview of species distributions, and characterisation of ecological niches & host records is still required to deeply understand the genus as a whole, yet our results already allow broad access to and inclusion of Anacharis species in downstream biodiversity research.

Our study aimed to develop an optimised laboratory protocol ensuring the preservation of morphological structures and extraction of high-quality DNA sequences from Psychodidae (Insecta, Diptera) specimens. With 310 analysed specimens, we investigated the impact of distinct laboratory treatments by employing two shaking categories (constant and interrupted) with five different incubation periods (16, 12, 8, 4 and 2 hours) during the DNA extraction process. Notably, 80.65% of the specimens exhibited morphological changes during DNA extraction. Our results indicated no statistical difference between constant and interrupted shaking for the total of morphological structures lost. However, within each shaking category, the loss of structures was influenced significantly by the incubation period. Prolonged incubation correlated with increased structural losses, whereas shorter incubation periods caused minor alterations in structures lost. In addition, our results showed a significant difference between constant and interrupted shaking treatments for DNA concentration. Likewise, the incubation period showed differences within each shaking category. Successful COI sequencing was achieved in 89.6% of specimens, with negligible differences in DNA fragment lengths across treatments. Our findings underscore the importance of an optimised protocol and its potential in systematic research involving nematoceran dipteran specimens by balancing morphological integrity and DNA extraction efficiency.

DNA barcodes are a great tool for accelerated species identification and for complementing species delimitation. Furthermore, DNA barcode reference libraries are the decisive backbone feature for any metabarcoding study in biodiversity monitoring, conservation or ecology. However, in some taxa, DNA barcodes cannot be generated with published primers at a satisfying success rate and these groups will consequently be largely missing from any barcoding-based species list. Here, we provide a custom DNA barcoding forward primer for the Eurytomidae (Hymenoptera, Chalcidoidea), elevating the success rate of high-quality DNA barcodes from 33% to 88%. Eurytomidae is a severely understudied, taxonomically challenging, species-rich group of primarily parasitoid wasps. High species numbers, diverse ecological roles and widespread and common presence identify Eurytomidae as one of many crucial families in terrestrial ecosystems. It is now possible to include Eurytomidae when studying and monitoring the terrestrial fauna, highlighting that barcoding-based approaches will need to routinely use different primers to avoid biases in their data and inferences. The new DNA barcoding protocol is also a prerequisite for our integrative taxonomy study of the group, aiming at delimiting and characterising Central European species and filling the GBOL (German Barcode Of Life) DNA barcode reference library with species-named and voucher-linked sequences.