When details matter: Integrative revision of Holarctic Coelophthinia Edwards (Diptera, Mycetophilidae), including mapping of its mitogenome, leads to the description of four new pseudocryptic species.

Background: The small genus Coelophthinia Edwards, 1941 of the subfamily Gnoristinae (Diptera, Mycetophilidae) is so far known to harbour four species from the Palaearctic, Nearctic and Neotropical Regions. Extensive DNA barcoding of fungus gnats of the family Mycetophilidae through the International Barcode of Life project (iBOL) have initiated integrative studies resulting in taxonomic upgrades and a better understanding of many species and their delimitation. The opportunity was also taken to describe the mitogenome of a member of Coelophthinia for the first time. New information: The integrative studies give evidence for splitting the European species C.thoracica Edwards, 1941 into three different species. Four new species are described from the USA, Japan and the Nordic Region in Europe, Coelophthiniacirra Kerr sp. n., Coelophthiniaitoae Kurina sp. n., Coelophthinialata Kjaerandsen sp. n. and Coelophthinialoraasi Kjaerandsen sp. n., raising the number of Holarctic species from two to six. The mitogenome of Coelophthinialoraasi sp. n. is described and analysed.

Current State of DNA Barcoding of Sciaroidea (Diptera)-Highlighting the Need to Build the Reference Library.

DNA barcoding has tremendous potential for advancing species knowledge for many diverse groups of insects, potentially paving way for machine identification and semi-automated monitoring of whole insect faunas. Here, I review the current state of DNA barcoding of the superfamily Sciaroidea (Diptera), a diverse group consisting of eight understudied fly families where the described species in the world makes up some 10% (≈16,000 species) of all Diptera. World data of Sciaroidea were extracted from the Barcode of Life online database BoldSystems (BOLD) and contrasted with results and experiences from a Nordic project to build the reference library. Well over 1.2 million (1,224,877) Sciaroidea specimens have been submitted for barcoding, giving barcode-compliant sequences resulting in 56,648 so-called barcode index numbers (BINs, machine-generated proxies for species). Although the BINs on BOLD already represent 3.5 times the number of described species, merely some 2850 named species (described or interim names, 5% of the BINs) currently have been assigned a BIN. The other 95% remain as dark taxa figuring in many frontier publications as statistics representing proxies for species diversity within a family. In the Nordic region, however, substantial progress has been made towards building a complete reference library, currently making up 55% of all named Sciaroidea BINs on BOLD. Another major source (31%) of named Sciaroidea BINs on BOLD comes from COI sequences mined from GenBank, generated through phylogenetic and integrative studies outside of BOLD. Building a quality reference library for understudied insects such as Sciaroidea requires heavy investment, both pre sequence and post sequence, by trained taxonomists to build and curate voucher collections, to continually improve the quality of the data and describe new species. Only when the BINs are properly calibrated by a rigorously quality-checked reference library can the great potential of both classical taxonomic barcoding, metabarcoding, and eDNA ecology be realized.

A molecular-based identification resource for the arthropods of Finland.

To associate specimens identified by molecular characters to other biological knowledge, we need reference sequences annotated by Linnaean taxonomy. In this study, we (1) report the creation of a comprehensive reference library of DNA barcodes for the arthropods of an entire country (Finland), (2) publish this library, and (3) deliver a new identification tool for insects and spiders, as based on this resource. The reference library contains mtDNA COI barcodes for 11,275 (43%) of 26,437 arthropod species known from Finland, including 10,811 (45%) of 23,956 insect species. To quantify the improvement in identification accuracy enabled by the current reference library, we ran 1000 Finnish insect and spider species through the Barcode of Life Data system (BOLD) identification engine. Of these, 91% were correctly assigned to a unique species when compared to the new reference library alone, 85% were correctly identified when compared to BOLD with the new material included, and 75% with the new material excluded. To capitalize on this resource, we used the new reference material to train a probabilistic taxonomic assignment tool, FinPROTAX, scoring high success. For the full-length barcode region, the accuracy of taxonomic assignments at the level of classes, orders, families, subfamilies, tribes, genera, and species reached 99.9%, 99.9%, 99.8%, 99.7%, 99.4%, 96.8%, and 88.5%, respectively. The FinBOL arthropod reference library and FinPROTAX are available through the Finnish Biodiversity Information Facility (www.laji.fi) at https://laji.fi/en/theme/protax. Overall, the FinBOL investment represents a massive capacity-transfer from the taxonomic community of Finland to all sectors of society.

Revision of the Exechiaparva group (Diptera: Mycetophilidae).

BACKGROUND: Exechia is a diverse genus of small fungus gnats, widespread in the Holarctic Region, while the fauna is largely unknown elsewhere, such as in the Afrotropical and Oriental Region. Members of Exechia can be arranged into several species groups, based on homologies in the male and female terminalia. The Exechiaparva group is delimited, based on male terminalia possessing a pair of gonocoxal lobes on the apicoventral gonocoxal margin. Eight previously-described species can be placed in this group, of which six are from the Holarctic Region, while one is recorded each from the Oriental and the Afrotropical Regions. NEW INFORMATION: The Exechiaparva group was reviewed and found to include 33 species, of which 24 were described as new to science and six were re-described. Identification keys to 32 species for males and nine species for females are provided together with illustrations and photos of male and female terminalia. Species delimitations were based on morphological examination of 94 male and female specimens, as well as DNA barcodes obtained from 124 specimens. Molecular and morphological species delimitations were mostly congruent, except in two cases where two species were delimited within a single Barcode Index Number (BIN). We found that each species is only known from a single zoogeographical region and that several species complexes are largely congruent with zoogeographical divisions, indicating that intercontinental barriers may have a strong impact on the species diversity of the group.

Coelosynapha, a new genus of the subfamily Gnoristinae (Diptera: Mycetophilidae) with a circumpolar, Holarctic distribution.

BACKGROUND: The subfamily Gnoristinae is one of the most diverse and taxonomically difficult subfamilies of Mycetophilidae, with new species and genera being described almost every year from various parts of the world. Through inventories of fungus gnats in the Nordic Region and Russia, a genus and species new to science was discovered, yet with links back to an illustration made by the late French entomologist Loïc Matile in the 1980s. DNA barcoding aligned it with yet another species new to science, distributed across Canada and documented through The Barcode of Life Data System (BOLD) by Paul D. N. Hebert and colleagues at the BOLD team. NEW INFORMATION: The new Holarctic genus, Coelosynapha gen. n. is described, consisting of two new species, the Palaearctic Coelosynapha loici sp. n. and the Nearctic Coelosynapha heberti sp. n. DNA-barcodes assign the two new species to distinctly separated (8.27% p-distance) Barcode Index Numbers (BINs) which are most closely aligned to unidentified species of Mycetophilidae from South Australia and Costa Rica on BOLD. The new genus shows morphological characteristics in between the two Holarctic genera Coelosia Winnertz, 1864 and Synapha Meigen, 1818 and further shows affinity to the southern continents genus Austrosynapha Tonnoir, 1929. The Palaearctic Coelosynapha loici sp. n., for which habitat requirements are best documented, is largely restricted to pristine, old-growth conifer (mostly spruce, Picea abies ssp. obovata) forests within the boreal vegetation zone, although it is also recorded from hummock tundra along the Anadyr River in Far East Russia.

Hidden in Plain Sight: Comprehensive Molecular Phylogeny of Keroplatidae and Lygistorrhinidae (Diptera) Reveals Parallel Evolution and Leads to a Revised Family Classification.

We provide the first molecular phylogeny of Keroplatidae and Lygistorrhinidae, families of fungus gnats (Diptera: Bibionomorpha: Sciaroidea). Phylogenies reconstructed by Maximum Likelihood and Bayesian methods, based on four nuclear and four mitochondrial gene markers (5106 base pairs) sequenced for 75 genera and 105 species, show Keroplatidae as monophyletic only with the family Lygistorrhinidae included, herewith treated as the subfamily Lygistorrhininae stat. nov. The subfamily Arachnocampinae is retained in the family, although lowering its overall support. An early branching clade, comprising species of Platyura Meigen, 1803 and Paleoplatyura melanderi Fisher, 1941, forms subfamily Platyurinae Loew, 1850 stat. nov. The subfamilies Sciarokeroplatinae and Macrocerinae grouped together with three genera considered here as Keroplatidae incertae sedis. Subfamily Lygistorrhininae forms a sister clade to subfamily Keroplatinae, both retained monophyletic with high support. The traditional division of the subfamily Keroplatinae into the tribes Orfeliini and Keroplatini appears as outdated, resting largely on adaptive characters prone to parallel evolution. We find support for an alternative tribe corresponding to the Cloeophoromyia-Asindulum genus group, but a tribal reclassification of the Keroplatinae is left for future studies. The genus Heteropterna Skuse, 1888 is considered as identical with Ctenoceridion Matile, 1972 syn. nov.

Completing Linnaeus's inventory of the Swedish insect fauna: Only 5,000 species left?

Despite more than 250 years of taxonomic research, we still have only a vague idea about the true size and composition of the faunas and floras of the planet. Many biodiversity inventories provide limited insight because they focus on a small taxonomic subsample or a tiny geographic area. Here, we report on the size and composition of the Swedish insect fauna, thought to represent roughly half of the diversity of multicellular life in one of the largest European countries. Our results are based on more than a decade of data from the Swedish Taxonomy Initiative and its massive inventory of the country's insect fauna, the Swedish Malaise Trap Project The fauna is considered one of the best known in the world, but the initiative has nevertheless revealed a surprising amount of hidden diversity: more than 3,000 new species (301 new to science) have been documented so far. Here, we use three independent methods to analyze the true size and composition of the fauna at the family or subfamily level: (1) assessments by experts who have been working on the most poorly known groups in the fauna; (2) estimates based on the proportion of new species discovered in the Malaise trap inventory; and (3) extrapolations based on species abundance and incidence data from the inventory. For the last method, we develop a new estimator, the combined non-parametric estimator, which we show is less sensitive to poor coverage of the species pool than other popular estimators. The three methods converge on similar estimates of the size and composition of the fauna, suggesting that it comprises around 33,000 species. Of those, 8,600 (26%) were unknown at the start of the inventory and 5,000 (15%) still await discovery. We analyze the taxonomic and ecological composition of the estimated fauna, and show that most of the new species belong to Hymenoptera and Diptera groups that are decomposers or parasitoids. Thus, current knowledge of the Swedish insect fauna is strongly biased taxonomically and ecologically, and we show that similar but even stronger biases have distorted our understanding of the fauna in the past. We analyze latitudinal gradients in the size and composition of known European insect faunas and show that several of the patterns contradict the Swedish data, presumably due to similar knowledge biases. Addressing these biases is critical in understanding insect biomes and the ecosystem services they provide. Our results emphasize the need to broaden the taxonomic scope of current insect monitoring efforts, a task that is all the more urgent as recent studies indicate a possible worldwide decline in insect faunas.

Allodia Winnertz from the Himalayas, with nine species new to science (Diptera, Mycetophilidae).

An extensive collection of fungus gnats from Nepal and Bhutan, deposited at Kyushu University Museum in Japan, has been examined and revealed nine species of Allodia Winnertz, 1864 new to science: Allodiacaligata Magnussen, sp. n., A.dibolia Magnussen, sp. n., A.shimai Magnussen, sp. n., A.spathulata Magnussen, sp. n., A.horologia Magnussen, sp. n., A.himalayensis Magnussen, sp. n., A.nepalensis Magnussen, sp. n., A.thudamensis Magnussen, sp. n., and A.scalprata Magnussen, sp. n. All specimens were collected at high altitudes in the central and eastern Himalayas. The species all belong to the subgenusAllodia s. str. and constitute the first records of the genus Allodia in Nepal and Bhutan. Brevicornunigrofasciatum (Brunetti, 1912) comb. n., originally described from northern India, is transferred from Allodia based on the original description. A key for the identification of the new species is provided.

Six new species of Afrotropical Allodia (Diptera: Mycetophilidae): DNA barcodes indicate recent diversification with a single origin.

Only one species of the genus Allodia has been previously recorded from the Afrotropical region, Allodia (Brachycampta) flavorufa Matile, 1978. Six new species are described here, all representing the nominotypical subgenus, Allodia s.s. The new species are described from material collected in different mountainous areas in south and east Africa; A. jaschhofi sp. nov., A. karkloofensis sp. nov., A. drakensbergensis sp. nov., A. nyeriensis sp. nov., A. mazumbaiensis sp. nov. and A. keurbosensis sp. nov. The species are morphologically very similar, and can only be separated based on minor differences in wing venation and characters of the male terminalia. The genetic differences between the species in the DNA barcode region (CO1), however, support delimitation. The origin and distribution of these Afrotropical taxa, in relation to each other and to their Holarctic relatives, is discussed.

Sexual selection on wing interference patterns in Drosophila melanogaster.

Animals with color vision use color information in intra- and interspecific communication, which in turn may drive the evolution of conspicuous colored body traits via natural and sexual selection. A recent study found that the transparent wings of small flies and wasps in lower-reflectance light environments display vivid and stable structural color patterns, called "wing interference patterns" (WIPs). Such WIPs were hypothesized to function in sexual selection among small insects with wing displays, but this has not been experimentally verified. Here, to our knowledge we present the first experimental evidence that WIPs in males of Drosophila melanogaster are targets of mate choice from females, and that two different color traits--saturation and hue--experience directional and stabilizing sexual selection, respectively. Using isogenic lines from the D. melanogaster Genetic Reference Panel, we compare attractiveness of different male WIPs against black and white visual backgrounds. We show that males with more vivid wings are more attractive to females than are males with dull wings. Wings with a large magenta area (i.e., intermediate trait values) were also preferred over those with a large blue or yellow area. These experimental results add a visual element to the Drosophila mating array, integrating sexual selection with elements of genetics and evo-devo, potentially applicable to a wide array of small insects with hyaline wings. Our results further underscore that the mode of sexual selection on such visual signals can differ profoundly between different color components, in this case hue and saturation.

Katatopygia gen. n., a monophyletic branch segregated from Boletina (Diptera, Mycetophilidae).

The genus Katatopygiagen. n. is proposed for the Boletina erythropyga/punctus-group that was first introduced by Garrett (1924, 1925) and currently comprises eight described species. Molecular studies have strongly indicated that this group forms a monophyletic sister-group to a clade consisting of all other Boletina, Coelosia and Gnoriste, and its monophyly is supported by morphological data as well. The new genus includes the following species: Katatopygia antoma (Garrett, 1924), comb. n., Katatopygia antica (Garrett, 1924), comb. n., Katatopygia erythropyga (Holmgren, 1883), comb. n.,Katatopygia hissarica (Zaitzev & Polevoi, 2002), comb. n., Katatopygia magna (Garrett, 1925), comb. n., Katatopygia laticauda (Saigusa, 1968), comb. n., Katatopygia neoerythropyga (Zaitzev & Polevoi, 2002), comb. n. andKatatopygia sahlbergi (Lundström, 1906), comb. n., all transferred from Boletina. Katatopygia sahlbergi is found to be a senior synonym of Boletina punctus Garrett, 1925, syn. n. A phylogeny based on morphological data and using parsimony analysis yielded four most parsimonious trees where the new genus is retrieved as monophyletic with high support. Katatopygia neoerythropyga is found to be the sister-taxon to all other species that form two clades, one with Katatopygia sahlbergi-like species and one with Katatopygia erythropyga-like species. A key to males of Katatopygia is provided.

Review of the European Greenomyia Brunetti (Diptera, Mycetophilidae) with new descriptions of females.

The females of the four continental Greenomyia Brunetti species in Europe are associated with the males, diagnosed and keyed, providing the first association and description of the females of Greenomyia baikalica Zaitzev, 1994 and Greenomyia stackelbergi Zaitzev, 1982. Colour photographs of their habitus and line drawings of their female terminalia are provided. Greenomyia mongolica Lastovka & Matile, 1974 is found to be a senior synonym of Greenomyia theresae Matile, 2002. syn. n. The diagnostic characters used to distinguish between Greenomyia and Neoclastobasis Ostroverkhova in keys did not hold up to a closer scrutiny and leave the status of Neoclastobasis as separate genus questionable.

Stable structural color patterns displayed on transparent insect wings.

Color patterns play central roles in the behavior of insects, and are important traits for taxonomic studies. Here we report striking and stable structural color patterns--wing interference patterns (WIPs)--in the transparent wings of small Hymenoptera and Diptera, patterns that have been largely overlooked by biologists. These extremely thin wings reflect vivid color patterns caused by thin film interference. The visibility of these patterns is affected by the way the insects display their wings against various backgrounds with different light properties. The specific color sequence displayed lacks pure red and matches the color vision of most insects, strongly suggesting that the biological significance of WIPs lies in visual signaling. Taxon-specific color patterns are formed by uneven membrane thickness, pigmentation, venation, and hair placement. The optically refracted pattern is also stabilized by microstructures of the wing such as membrane corrugations and spherical cell structures that reinforce the pattern and make it essentially noniridescent over a large range of light incidences. WIPs can be applied to map the micromorphology of wings through direct observation and are useful in several fields of biology. We demonstrate their usefulness as identification patterns to solve cases of cryptic species complexes in tiny parasitic wasps, and indicate their potentials for research on the genetic control of wing development through direct links between the transregulatory wing landscape and interference patterns we observe in Drosophila model species. Some species display sexually dimorphic WIPs, suggesting sexual selection as one of the driving forces for their evolution.

Blood-feeding behavior of Anopheles gambiae and Anopheles melas in Ghana, western Africa.

Anopheles gambiae is the predominant malaria vector species in Ghana, western Africa, with a strong local presence of Anopheles melas Theobald along the southern coast. We studied the biting behavior of these two species of the Anopheles gambiae complex inland and at the coast in Ghana, with special attention to the local peoples' preference for outdoor sleeping. We collected mosquitoes at two sites in 2007, representing the moist semideciduous forest zone and the strand and mangrove zone, and the sampling was repeated in the dry and rainy seasons. Sampled mosquitoes were examined for species, parity and size (wing length), and we identified the hosts of their bloodmeals. We interviewed 288 of the village people to determine where and when they slept outdoors. Our study confirmed that An. gambiae is the only species of the An. gambiae complex in the Ashanti region and revealed that An. melas is highly dominant on the western coast of Ghana. Both species showed high human blood rates in indoor resting mosquito samples. More people sleep outside on the coast than inland. An. melas demonstrated high exophily. An. gambiae bit people more frequently indoors and did so more often during the dry season than in the rainy season. We suggest that the degree of exophily in An. melas may be affected by humidity and the availability of human as well as by the mosquitoes' innate habits.