Odorant-binding Protein 10 From Bradysia odoriphaga (Diptera: Sciaridae) Binds Volatile Host Plant Compounds.

Bradysia odoriphaga (Diptera: Sciaridae) is a major insect pest of seven plant families including 30 commercial crops in Asia. The long-term use of chemical pesticides leads to problems such as insect resistance, environmental issues, and food contamination. Against this background, a novel pest control method should be developed. In insects, odorant-binding proteins (OBPs) transport odor molecules, including pheromones and plant volatiles, to olfactory receptors. Here, we expressed and characterized the recombinant B. odoriphaga OBP BodoOBP10, observing that it could bind the sulfur-containing compounds diallyl disulfide and methyl allyl disulfide with Ki values of 8.01 µM and 7.00 µM, respectively. Homology modeling showed that the BodoOBP10 3D structure was similar to that of a typical OBP. Both diallyl disulfide and methyl allyl disulfide bound to the same site on BodoOBP10, mediated by interactions with six hydrophobic residues Met70, Ile75, Thr89, Met90, Leu93, and Leu94, and one aromatic residue, Phe143. Furthermore, silencing BodoOBP10 expression via RNAi significantly reduced the electroantennogram (EAG) response to diallyl disulfide and methyl allyl disulfide. These findings suggest that BodoOBP10 should be involved in the recognition and localization of host plants.

Population genetic structure of a recent insect invasion: a gall midge, Asynapta groverae (Diptera: Cecidomyiidae) in South Korea since the first outbreak in 2008.

Outbreaks of Asynapta groverae, an invasive mycophagous gall midge, in South Korea have been repeatedly reported since the first occurrence in 2008. This species is a nuisance to residents owing to its mass emergence from newly built and furnished apartments. Here, the levels of genetic diversity, divergence, and structure of invasive A. groverae populations were investigated to understand their ability to survive in novel locations. Population genetic analyses were performed on seven invasive populations, including the first outbreak, sporadically emerged, and two laboratory-isolated (quarantined) populations, using the mitochondrial COI sequences and the ten novel microsatellite markers developed in this study. Non-indigenous A. groverae managed to maintain their populations for 12 years despite decreased genetic polymorphisms resulting from multiple incidences of founder effects by a small number of colonists. Additionally, the advantageous sustainability of A. groverae in the particle boards from which they emerge suggests that human-mediated dispersal is plausible, which may allow for the successful spread or invasion of A. groverae to new locations. This study is one of the few examples to demonstrate that an insect species successfully invaded new regions despite exhibiting decreased genetic diversity that was maintained for a decade. These findings indicate that the high genetic diversity of the initial founding population and asexual reproduction would contribute to the successful invasion of A. groverae in novel environments.

The oldest Gondwanan fossil of Leiinae (Diptera, Mycetophilidae): Phylogenetic and evolutionary implications.

A fossil Mycetophilidae from the Aptian Crato Formation-Cretomanota gondwanica gen. nov., sp. nov.-is described, which is the first mycetophilid from the Crato Formation and corresponds to the oldest known fossil leiine and only the second Gondwanan fossil mycetophilid described so far. Cretomanota gondwanica and both species of Alavamanota Blagoderov and Arillo were added as terminals to the data matrix of a general phylogenetic analysis of the Mycetophilidae, and both fit into the Leiinae. Alavamanota is monophyletic, sister to the clade composed by Cretomanota and the extant genus Manota Williston. The biology of the extant members of this fungivorous family corroborates the reconstruction of the Crato palaeoenvironment as including woodlands with humid habitats and microhabitats. The presence of a Cretaceous member of the tribe Manotini at low latitudes in South America reinforces the hypothesis that the clade with all manotines except Leiella Edwards corresponds to a Lower Cretaceous offshoot from a group in southern Gondwana expanding its distribution to more northern areas into the Gondwana and into Laurasia.

New finding of Toxorhina (Ceratocheilus) limoniid fly in Eocene Baltic amber and the biogeographical context of the genus.

Based on new fossil materials, a new species Toxorhina (Ceratocheilus) christelius sp. nov. has been described herein with complete documentation of drawings and photographs. Features such as wide spine on the gonocoxite differentiating the new species of Toxorhina were discussed. Finding new interesting fossil materials also allowed for providing an emended diagnosis and additional description of known Eocene species-Toxorhina (Ceratocheilus) eridanus. Comparison of chosen morphological features of fossil and recent representatives of the genus were given and key for fossil species of subgenus Ceratocheilus was introduced. Distribution of recent Toxorhina and evolutionary history of the genus were discussed. The results of research on fossil materials prove that the stratigraphic range of the subgenus Ceratocheilus and the genus Toxorhina goes back to the Eocene, there is no evidence of their existence on Earth before. Moreover, these insects were probably associated with a warm climate, they were found for example in Baltic amber, the deposits of which were formed mainly in a subtropical climate. The reach diversity of the genus Toxorhina of recent fauna is strictly observed in tropical zones around the world.

Arboviruses in Free-Ranging Birds and Hematophagous Arthropods (Diptera, Nematocera) from Forest Remnants and Urbanized Areas of an Environmental Protection Area in the Amazon Biome.

The rapid and disorderly urbanization in the Amazon has resulted in the insertion of forest fragments into cities, causing the circulation of arboviruses, which can involve hematophagous arthropods and free-ranging birds in the transmission cycles in urban environments. This study aimed to evaluate the circulation of arboviruses in free-ranging birds and hematophagous arthropods captured in an Environmental Protection Area in the Belem metropolitan area, Brazil. Birds were captured using mist nets, and hematophagous arthropods were collected using a human protected attraction technique and light traps. The birds' sera were subjected to a hemagglutination inhibition test to detect antibodies against 29 arbovirus antigens. Arthropod macerates were inoculated into C6/36 and VERO cell cultures to attempt viral isolation and were tested using indirect immunofluorescence, subsequent genetic sequencing and submitted for phylogenetic analysis. Four bird sera were positive for arbovirus, and one batch of Psorophora ferox was positive for Flavivirus on viral isolation and indirect immunofluorescence. In addition, the Ilheus virus was detected in the sequencing and phylogenetic analysis. The presence of antibodies in sera from free-ranging birds and the isolation of Ilheus virus in Psorophora ferox indicate the circulation of arboviruses in forest remnants in the urban center of Belem.

A new species of Hyperlasion Schmitz (Diptera: Sciaridae), causing periodic outbreaks in Japan.

A sciarid species, Hyperlasion breviantenna sp. n., is described from Japan. This is the first record of Hyperlasion Schmitz, 1918, from Asia. We compared the molecular sequence data of the mitochondrial cytochrome c oxidase subunit I (COI) region and external morphological characters among congeners and related genera. Morphological features are described and illustrated, and genetic relatedness to selected species with a one-segmented maxillary palpus is shown as a maximum likelihood tree. The DNA barcoding approach revealed that the genetic sequences of Japanese specimens are identical with those of Australian specimens, which have been assigned to the genus Hyperlasion. The new species occurs in outbreaks during the rainy season, June to July, in Japan and is recognized as a nuisance pest. Newly emerged adults appear in the early morning and enter houses, facilities, and public buildings. The biology of the new species is compared with those of H. wasmanni Schmitz, 1918, and Moehnia erema Pritchard, 1960, which have been recorded as occurring in large aggregations with thousands of individuals abroad, based on published biological notes and reports on these species.

Four genera of Limoniidae (Diptera) new to Japan with descriptions of new species.

Four limoniid genera are added to Japan for the first time. Four new species, Crypteria nippon sp. nov., Dasymallomyia tachii sp. nov., Eupilaria japonica sp. nov., and Protohelius japonicus sp. nov. are described, and Dasymallomyia ditenostyla Alexander, 1964 is recorded from Japan for the first time. Images of the habitus and wings and drawings of the male and female terminalia are provided.

Two new species of Setostylus Matile, 1990 (Diptera: Keroplatidae) from China.

In this study, two new species of Setostylus (Diptera: Keroplatidae: Keroplatinae), S. tridigitus sp. n. and S. triumphus sp. n. are described, with a key to all the species of the genus. Male habitus and images of diagnostic morphological characteristics are provided. Status of these two new species is also supported by the genetic distances and neighbor-joining (NJ) tree in the DNA barcode analysis.

Redescription of Dentifibula viburni (Felt) (Diptera: Cecidomyiidae) and review of the genus.

Dentifibula viburni (Felt) (Diptera: Cecidomyiidae), type species of Dentifibula Felt, is redescribed with illustrations and compared with its congeners. The genus is differentiated from the generally similar Lestodiplosis Kieffer. As far as presently known, larvae of Dentifibula prey on Diaspididae (Hemiptera: Coccoidea). The nine other species of Dentifibula are diagnosed. Dentifibula ceylanica Felt is removed from synonymy under Dentifibula obtusiloba Felt.

Development of Transformation for Genome Editing of an Emerging Model Organism.

With the advances in genomic sequencing, many organisms with novel biological properties are ripe for use as emerging model organisms. However, to make full use of them, transformation methods need to be developed to permit genome editing. Here, we present the development of transformation for the fungus fly Bradysia (Sciara) coprophila; this may serve as a paradigm for the development of transformation for other emerging systems, especially insects. Bradysia (Sciara) has a variety of unique biological features, including locus-specific developmentally regulated DNA amplification, chromosome imprinting, a monopolar spindle in male meiosis I, non-disjunction of the X chromosome in male meiosis II, X chromosome elimination in early embryogenesis, germ-line-limited (L) chromosomes and high resistance to radiation. Mining the unique biology of Bradysia (Sciara) requires a transformation system to test mutations of DNA sequences that may play roles for these features. We describe a Bradysia (Sciara) transformation system using a modified piggyBac transformation vector and detailed protocols we have developed to accommodate Bradysia (Sciara) specific requirements. This advance will provide a platform for us and others in the growing Bradysia (Sciara) community to take advantage of this unique biological system. In addition, the versatile piggyBac vectors described here and transformation methods will be useful for other emerging model systems.

Cretaceous Antodicranomyia (Diptera: Limoniidae) and their paleohabitat.

New representatives of the Cretaceous cranefly genus Antodicranomyia (Diptera: Limoniidae) are reported from Albian-Cenomanian Charentese (French) amber. The newly reported specimens allow for an emended diagnosis of the type species A. azari, as well as the description of a new species, Antodicranomyia rubra sp. nov., which is mostly distinguished from the type species by features of its wing venation, antennae, and genitalia. As a rare, extinct genus known only from French amber, Antodicranomyia is compared with its closest relative genera Antocha, Dicranomyia and Antohelia. The evolutionary implications and paleohabitat of Antodicranomyia are discussed. The new discovery adds to the knowledge of the crane flies' diversity and evolution in the mid-Cretaceous.

Tipula (Vestiplex) levisoni sp. nov., a new cranefly (Diptera, Tipulidae) from the Western Himalayas.

A new cranefly, Tipula (Vestiplex) levisoni sp. nov., is described and illustrated based on males and females collected in India and Nepal. The illustrations of male and female genitalia for the most closely related regional species, T. (V.) tanycera Alexander, 1961 and T. (V.) mitchelli Edwards, 1927 are also provided. The female of T. (V.) mitchelli is described for the first time.

A new giant dung midge from Miocene Ethiopian amber (Diptera: Scatopsidae).

Fossil new genus and species Afroscatopse haennii gen. et sp. nov. from Miocene Ethiopian amber are described and illustrated and placed in the tribe Colobostematini. The gigantic adults of this scavenger fly, largest within the family, with dull body, long antennae and broad wings evidently were adapted to live on bark of trees. It is suggested that it was a myrmecophilous species with larvae living in the nests of tree ants.

A new species of Chetoneura Colless from Borneo (Diptera: Keroplatidae).

A new species of KeroplatidaeChetoneura lagangnsis sp. n. from Lagang cave in Sarawak, Borneo is described and illustrated based on a single male specimen. An identification key to the species of the genus Chetoneura Colless is provided. A distribution map of the five known species of Chetoneura is presentedCh. cavernae Colless, Ch. shennonggongensis Amorim Niu, Ch. oligoradiata (Papp), Ch. davidi evk, Hippa Burdikov and Ch. lagangensis Kazandzhieva Bechev, sp. n.

An unknown world in the Neotropical region: a complete life cycle of a new species of Monoclona Mik, 1886 (Diptera: Mycetophilidae: Sciophilinae).

Popularly known as fungus gnats, Mycetophilidae are found in humid environments usually associated with mushrooms and decaying wood. Their immature forms often feed on fungus fruiting bodies. Similar to most bibionomorphans, mycetophilids need due attention concerning their taxonomy and information on their natural history, especially in the Neotropical region. This work describes Monoclona carambeiensis sp. nov., a new species of Monoclona Mik, and furnishes information on the morphology of adults and immatures, besides notes and photographs on the life cycle of the species. Immature forms present on a piece of decaying wood with lichens and fungi were collected from Carambe, Paran, and reared in the laboratory. The emerged adult male was fixed in 80% ethanol. This is the first study describing an immature of a Neotropical species of Monoclona, and also the first record of the genus in the state of Paran, Brazil. This is the third Neotropical species of Mycetophilidae to have its life cycle described, for a fauna with over 1,100 known species.

A new PCR based molecular method for early and precise quantification of parasitization in the emerging olive pest Dasineura oleae.

BACKGROUND: Dasineura oleae (Angelini 1831) (Diptera: Cecidomyiidae) was considered a minor pest in olive orchards, but in recent years severe outbreaks have been registered in several Mediterranean countries. Damage is caused by the feeding activity of larvae that induce gall formations and alters the physiological activity of the leaves. In Italy, this pest may be controlled by four Hymenoptera parasitoid species belonging to Platygaster and Mesopolobus genera such as Platygaster demades Walker 1835, Platygaster oleae Szelenyi 1940 (Hymenoptera: Platygastridae), Mesopolobus aspilus (Walker 1835) and Mesopolobus mediterraneus (Mayr 1903) (Hymenoptera: Pteromalidae), but parasitization becomes evident only after gall dissection. RESULTS: In this study, we aim to: (i) design a primer for the detection of specimens belonging to Platygaster and Mesopolobus genera; (ii) develop a multiplex quantitative polymerase chain reaction (qPCR) protocol combined to a fast samples DNA extraction method; (iii) apply the developed protocol to field-collected specimens and compare this method with traditional techniques based on visual estimation of parasitism rate on larvae. Primers were designed to anneal with cytochrome oxidase subunit I (COI) sequences of Platygaster and Mesopolobus genera while protocols were developed to be fast and capable to process several samples at the same time. Molecular analyses demonstrated to provide almost double of the parasitism rate assessed by visual inspection. Furthermore, on second instar larvae the PCR-based method was able to detect ten-fold times the parasitization rate estimated by visual inspection. CONCLUSION: The application on a greater scale of this newly developed method could be fundamental in the determination of the biological control potential in olive orchards.

Characterization of the insecticide detoxification carboxylesterase Boest1 from Bradysia odoriphaga Yang et Zhang (Diptera: Sciaridae).

BACKGROUND: In insects, carboxylesterases (CarEs) are enzymes involved in the detoxification of insecticides. However, the molecular mechanism of CarE-mediated insecticide metabolism in Bradysia odoriphaga, a serious agricultural pest, remains unclear. The aim of this study is to investigate the detoxification process of malathion, bifenthrin, and imidacloprid by B. odoriphaga carboxylesterase (Boest1). RESULTS: An alpha class CarE gene Boest1 was cloned from B. odoriphaga. The results of real-time quantitative polymerase chain reaction showed that Boest1 is up-regulated with age during the larval stage, and the level of transcription of Boest1 is higher in the midgut and Malpighian tubule than in other tissues. The expression level of Boest1 was significantly increased after exposure to malathion and bifenthrin. Recombinant BoEST1 expressed in vitro showed high catalytic activity toward α-naphthyl acetate, which was substantially inhibited by malathion and triphenyl phosphate. The in vitro metabolism assays showed that BoEST1 demonstrates hydrolytic capacity toward malathion and bifenthrin but not imidacloprid. The binding free energy analysis indicates that BoEST1 has a higher affinity for malathion and bifenthrin than imidacloprid. CONCLUSION: These results suggest that BoEST1 plays a role in the breakdown of insecticides and may be involved in the development of resistance in the Chinese chive pest B. odoriphaga; our findings also provide data for better pest management and perspectives for new pesticides development. © 2021 Society of Chemical Industry.

Taxonomic study of the genus Eloeophila Rondani, 1856 of Japan (Limoniidae, Diptera).

Japanese species of the genus Eloeophila are revised. Five new species, E. apicisetula sp. nov., E. canidorsalis sp. nov., E. enischnophallus sp. nov., E. hadrophallus sp. nov., and E. tergilobellus sp. nov. are described. One subspecies and two species are synonymized, E. subaprilina yezoensis (Alexander, 1924) with the nominotypical subspecies, E. hidana (Alexander, 1970) with E. persalsa (Alexander, 1940), and E. mishimai (Alexander, 1969) with E. kintaro (Alexander, 1957). Images of the habitus and wings, and drawings of the male terminalia of the Japanese species are shown. Distributions and a key to the Japanese species are provided.

When unexpected guests come to eat: two new species of Phyllolabis Osten Sacken (Diptera: Limoniidae) collected with carrion-baited traps in the centre of the Iberian Peninsula.

Phyllolabis eiroae sp. nov. and P. martinhalli sp. nov. are described from the centre of the Iberian Peninsula. These two remarkable species were collected using carrion-baited traps, running during winter, in several localities of Madrid province (Spain). The two new species are well differentiated from the other Phyllolabis Osten Sacken species recorded from the Iberian Peninsula, P. savtshenkoi Theowald, and those from the west Palaearctic. An identification key to differentiate the three Phyllolabis species occurring in the Iberian Peninsula is provided. The first images of P. savtshenkoi, based on the holotype and a male specimen recorded from a cave located in Ja (Spain), are also provided.