Express barcoding with NextGenPCR and MinION for species-level sorting of ecological samples.

The use of DNA barcoding is well established for specimen identification and large-scale biodiversity discovery, but remains underutilized for time-sensitive applications such as rapid species discovery in field stations, identifying pests, citizen science projects, and authenticating food. The main reason is that existing express barcoding workflows are either too expensive or can only be used in very well-equipped laboratories by highly-trained staff. We here show an alternative workflow combining rapid DNA extraction with HotSHOT, amplicon production with NextGenPCR thermocyclers, and sequencing with low-cost MinION sequencers. We demonstrate the power of the approach by generating 250 barcodes for 285 specimens within 6 h including specimen identification through BLAST. The workflow required only the following major equipment that easily fits onto a lab bench: Thermocycler, NextGenPCR, microplate sealer, Qubit, and MinION. Based on our results, we argue that simplified barcoding workflows for species-level sorting are now faster, more accurate, and sufficiently cost-effective to replace traditional morpho-species sorting in many projects.

Taxonomic reappraisal of the European fauna of the bark beetle genus Cryphalus (Coleoptera, Curculionidae, Scolytinae).

Species in the genus Cryphalus are small and notoriously difficult to identify. Even among the relatively well studied European species, erroneous identifications are evident from literature and in museum collections. These misidentifications relate to the small size and similar appearance of Cryphalus species but they are also a product of insufficient diagnostic characters. This is especially unfortunate since some European species are considered pests. Based on the study of more than 1000 specimens and a thorough literature review, robust morphological and molecular evidence supporting all five hitherto recognised native species of Cryphalus in Europe is provided. A key for the reliable identification of these repetition species including new diagnostic characters recognised for the first time, including those from male genitalia, has been constructed. Each native species is provided with a detailed morphological description and their economic significance, distribution, and ecology discussed. Significant genetic variability is observed between certain clusters that should be further explored in a broader geographic context. Lastly, the need for a taxonomic revision of the genus Cryphalus for the entire Palearctic region due to the presence of many similar looking species which are often confused, thus distorting the knowledge of each species is highlighted.

Formation of the Holarctic Fauna: Dated molecular phylogenetic and biogeographic insights from the Quedius-lineage of Ground-Dwelling Rove Beetles (Coleoptera, Staphylinidae).

Although the Holarctic fauna has been explored for centuries, many questions on its formation are still unanswered. For example, i) what was the impact of the uplift of the Himalaya and Tibetan Plateau?, ii) what were the timings and climate of the faunal bridges connecting the Nearctic and Palearctic regions?, and iii) how did insect lineages respond to the late Paleogene global cooling and regional aridification? To answer these, we developed a phylogenetic dataset of 1229 nuclear loci for a total of 222 species of rove beetles (Staphylinidae) with emphasis in the tribe Quediini, especially Quedius-lineage and its subclade Quedius sensu stricto. Using eight fossils for calibrating molecular clock, we estimated divergence times and then analysed in BioGeoBEARS paleodistributions of the most recent common ancestor for each target lineage. For each species we generated climatic envelopes of the temperature and precipitation and mapped them across the phylogeny to explore evolutionary shifts. Our results suggest that the warm and humid Himalaya and Tibetan Plateau acted as an evolutionary cradle for the Quedius-lineage originating during the Oligocene from where, in the Early Miocene, the ancestor of the Quedius s. str. dispersed into the West Palearctic. With the climate cooling from the Mid Miocene onwards, new lineages within Quedius s. str. emerged and gradually expanded distributions across the Palearctic. In Late Miocene, a member of the group dispersed to the Nearctic region via Beringia before the closure of this land bridge 5.3 Ma. Paleogene global cooling and regional aridification largely shaped the current biogeographic pattern for Quedius s. str. species, many of them originating during the Pliocene and shifting or contracting their ranges during Pleistocene.

A New Termitophilous Genus of Paederinae Rove Beetles (Coleoptera, Staphylinidae) from the Neotropics and Its Phylogenetic Position.

We describe a new genus and species of Paederinae rove beetles, Ruptor cordatus gen. et sp. nov., which lives in the arboreal nests of the termite Labiotermes labralis (Holmgren, 1906) in the Amazon lowlands of Peru. The morphology of Ruptor gen. nov. is highly derived, apparently due to its close association with the termite host, and thus, morphologically, the genus cannot be classified further than Lathrobiini incertae sedis. In order to address the sister-group relationships of Ruptor gen. nov., we conducted a molecular phylogenetic analysis based on seven gene fragments. The analysis indeed resolved the genus as a member of the tribe Lathrobiini and placed it nested within the informal clade of 'Medonina and allied taxa'. We provide a morphological comparison of the new genus with all known myrmeco- and termitophilous representatives of the subfamily, and to the extent possible, we illustrate other relevant and poorly known Neotropical Paederinae inquilines.

Checklist and taxonomic changes for Central and South American Philonthina (Coleoptera: Staphylinidae).

A checklist of all described species of Philonthina, a subtribe of the staphylinid tribe Staphylinini, known to occur in Central and South America (CASA) is presented. Included for each species, and for synonyms known from CASA, is a reference to the original description, type locality and type depository, and for each species the known distribution within and outside CASA. Type material was sought in the main European and American collections where it is deposited (BMNH, MNHUB, IRSNB and FMNH) and is summarized for all indigenous CASA species, with lectotypes designated for 16 names and confirmation of holotypes and prior designation of lectotypes when necessary. Based on recent phylogenetic work in Philonthina and our revision of types of CASA species of Philonthus Stephens, 1829 and Belonuchus Nordmann, 1837, some taxonomic changes are proposed. Thirty-one species of Philonthus are transferred to Belonuchus (16), Gabrius Stephens 1829 (14), and Bisnius Stephens 1829 (one) resulting in the following new combinations: B. abnormalis (Sharp 1885), B. celatus (Sharp 1885), B. corticalis (Sharp 1885), B. extremus (Sharp 1885), B. infimus (Sharp 1885), B. iteratus (Sharp 1887), B. latecinctus (Sharp 1885), B. lucilius (Sharp 1885), B. muticus (Sharp 1876), B. optatus (Sharp 1885), B. platypterus (Sharp 1885), B. rufiventris (Sharp 1887), B. rufocaudus (Sharp 1885), B. rufopygus (Sharp 1885), B. serraticornis (Sharp 1876), B. supernus (Herman 2001), G. approximans (Sharp 1885), G. armatipes (Sharp 1885), G. atricolor (Sharp 1885), G. championi (Sharp 1885), G. dampfi (Bernhauer 1929), G. elegans (Sharp 1885), G. forsterianus (Scheerpeltz 1960), G. misellus (Sharp 1885), G. nugax (Sharp 1885), G. ovaticeps (Sharp 1885), G. peruvianus (Bernhauer 1916), G. planulatus (Sharp 1885), G. rusticus (Sharp 1885), G. serpens (Sharp 1885) and Bi. subaeneipennis (Bernhauer 1916). Endeius nitidipennis Solier 1849 is transferred to Gabrius, resulting in the following new combination, G. nitidipennis (Solier 1849). Leptopeltus carchiensis Chani-Posse Asenjo 2013 is proposed as junior synonym of Philonthus divisus Sharp 1891, which is transferred to Leptopeltus Bernhauer 1906 resulting in a new combination: Leptopeltus divisus (Sharp 1891). Belonuchus penetrans Silvestri 1946 is transferred to Pridonius Blackwelder 1952 as a new combination. Lectotypes are designated for Atopocentrum mirabile Bernhauer 1906, Philonthus armatipes Sharp 1885, Ph. atricolor Sharp 1885, Ph. championi Sharp 1885, Ph. misellus Sharp 1885, Ph. planulatus Sharp 1885, Ph. rusticus Sharp 1885, Ph. serpens Sharp 1885, Ph. abnormalis Sharp 1885, Ph. celatus Sharp 1885, Ph. infimus Sharp 1885, Ph. latecinctus Sharp 1885, Ph. muticus Sharp 1876, Ph. platypterus Sharp 1885, Ph. rufocaudus Sharp 1885 and Ph. rufopygus Sharp 1885. Of the 543 currently known species of Philonthina reported from CASA, at least 14 are believed to be adventive from elsewhere, 56 may occur naturally elsewhere, and 473 (87%) are evidently endemic to this region. Of the 31 genera represented by these described species, 20 (65%) are endemic to CASA. One genus, Gabronthus Tottenham 1955, is adventive. However, the actual philonthine fauna of CASA will undoubtedly be much larger, and the generic composition highly modified, when the fauna is fully explored and studied within a phylogenetical framework.

Review of subterranean Quedius, with description of the first hypogean species from the Russian Far East (Coleoptera: Staphylinidae: Staphylinini).

Hypogean species of Quedius, one of the largest North Hemisphere beetle genera, are reviewed in connection with the discovery of a new peculiar, brachypterous and microphtalmous species from the subgenus Microsaurus in the talus-based microhabitats of the Russian Far East. Morphology of that species here described as Quedius roma sp. n. suggests its affinity with the przewalskii-group from China and Nepal. Affinity of Q. roma to the abnormalis-group, a geographically more proximate and species-rich and presumably non-monophyletic assemblage of hypogean species from Japan and Taiwan, or to any other Holarctic Microsaurus is not supported morphologically. Two hypogean species from Europe, Q. bernhaueri Rambousek, 1915 and Q. doderoi Gridelli, 1922, are transferred from the subgenus Microsaurus to the subgenus Raphirus. The name Tenebrobius Rambousek, 1915, a subgenus where Q. bernhaueri was originally described, becomes a junior synonym of Raphirus Stephens, 1829.