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In this work we report for the first time the occurrence of female adults of the biting midge Forcipomyia (Microhelea) eriophora feeding on butterfly Heraclides anchisiades capys caterpillars. We collected the caterpillars and Ceratopogonidae specimens on Citrus limon (L.) (Rutaceae) tree leaves from the campus of the Universidade Estadual de Feira de Santana (UEFS), Feira de Santana, Bahia, Brazil. In Brazil, host records of Forcipomyia (Microhelea) eriophora on caterpillars remain scarce due to significant gaps in our understanding of ectoparasitic habits within Ceratopogonidae.

Aims The main aim of this study was to fill a gap in barcoding data of the European Chironomidae of the Balkan region, developing and testing the efficiency of a reference DNA barcode library for ancient Lake Skadar basin species (Montenegro/Albania), a region of Europe never before subjected to barcoding studies on Chironomidae. Another aim was to test the efficiency of DNA barcoding for the identification of European Chironomidae, including the estimation of optimal identification thresholds, using >12,000 barcodes. Location Lake Skadar basin and adjacent area (Montenegro/Albania). Methods Through this study, 770 individuals of Chironomidae from the Lake Skadar region were barcoded, both at adult and pre‐imaginal stages. Adults were morphologically identified, while larvae were assigned to species by molecular identification, using different methods, of which the efficiency was tested, for a total of 97 different barcoded species. Results The identification efficiency of the reference dataset developed for the Lake Skadar region was 98.6%, a value in line with that obtained when the identification efficiency for European Chironomidae was evaluated (95.8%), which confirms the accuracy of DNA barcoding for the identification of these insects. Moreover, we found that the optimal threshold for the molecular identification of the family is 1.6% nucleotide distance, though more specific thresholds are suggested for the identification of species belonging to Chironomidae subfamilies, since they are related to lower identification errors than to the use of a general threshold. The analysis of inconsistency between molecular and morphological identification shed light on taxonomic issues within European Chironomidae. Previously postulated species synonyms were confirmed, and also further cases requiring deeper investigation were detected. Main conclusions Our de novo DNA barcode library was shown to have a high identification efficiency. Taxon‐specific thresholds increase the efficacy of molecular identification. Hypothesized species synonyms could be validated through molecular techniques.

In October 2023, bluetongue virus serotype 3 (BTV-3) emerged in Germany, where Schmallenberg virus is enzootic. We detected BTV-3 in 1 pool of Culicoides biting midges collected at the time ruminant infections were reported. Schmallenberg virus was found in many vector pools. Vector trapping and analysis could elucidate viral spread.

Culicoides (Diptera: Ceratopogonidae) Latreille biting midges are the vectors of several viruses of veterinary significance. The larvae of some biting midge species develop in semiaquatic habitats that are affected by the activity of vertebrates. However, the importance of vertebrate animal activity in biting midge density has not been satisfactorily quantified. The goal of this study was to investigate the impact that three different hooved animals representing agricultural (cattle), wild (cervids), and intermediate (bison) animal populations have on the density and emergence of Culicoides from larval habitats. Trail cameras recorded vertebrate activity at eight sites at the Konza Prairie Biological Station with cattle, cervids, and/or bison, which was paired with larval substrate sampling to investigate midge density. The total time spent by all 3 target animals over the previous month and the average time spent by bison were significantly positively correlated with overall biting midge emergence. Species-specific analyses revealed significant positive associations of C. crepuscularis Malloch with several cattle variables including number of cattle days and events and average number of cattle, while more bison variables were found to be significant for C. haematopotus Malloch (bison days and bison events, negative associations) and C. variipennis Coquillett (negative associations for bison days and average number of bison; positive relationship with average bison time). Significant results for cervids included negative associations of C. crepuscularis with average cervid time and C. variipennis with cervid days. These results show that different ungulates impact midge abundance and emergence in different ways, improving our understanding of midge population drivers.

Epizootic hemorrhagic disease virus (EHDV: Reoviridae: Orbivirus ) is a Culicoides- borne pathogen that affects a variety of ruminants, causing significant economic losses and/or ecological impacts in animal agriculture/wildlife populations worldwide. In this study, we examined the effect of EHDV serotype-2 oral infection on the survival and reproduction of Culicoides sonorensis Wirth and Jones (a confirmed vector of EHDV in North America), and the potential vertical transmission of EHDV-2 (from infected female to its offspring) in this midge species. Culicoides sonorensis females were fed on defibrinated bovine blood mixed with EHDV-2 (5.5 log 10 PFU/ml) or without EHDV-2 (control). Adult survival/longevity, oviposition rates, number of eggs deposited, egg hatch rates (fertility), larval survival, larval stage duration, eclosion rates, and sex-ratios of the progeny were recorded and compared between the two groups. In addition, the progeny (eggs and F 1 generation adults) of EHDV-2 fed females were processed for viral detection through RT-qPCR and plaque assays. Survival/longevity of the blood-fed adults, oviposition rates, number of eggs deposited, larval stage duration, eclosion rates, and sex-ratios were not significantly different between the two groups. However, egg hatch rates were significantly lower in the EHDV-2 fed group (35.8 ± 5.2%) than the control group (74.5 ± 6.8%), but larval survival rates were higher in the EHDV-2 fed group (59.8 ± 4.9%) compared to the control group (34.1 ± 6.5%). EHDV-2 (Ct < 35) was detected in the eggs (3.4%, 1/29 females tested, Ct = 22.1 [4.9 log 10 PFUe/ml]) and F 1 adult progeny (1.7%, 1/58 adults tested, Ct = 23.5 [4.5 log 10 PFUe/ml]) of the orally exposed females through RT-qPCR as well as through plaque assays. Our findings suggest that EHDV-2 infection has no major impact on C. sonorensis survival/longevity or oviposition but has a significant negative effect on midge fecundity/fertility. Our study also provides evidence for the vertical transmission of EHDV-2 from an infected adult female to its offspring in C. sonorensis . However, salivary transmission of EHDV-2 from the vertically infected progeny and its significance in the epidemiology of hemorrhagic disease are currently unknown and remain to be examined in further studies. Overall, these findings collectively indicate that Orbivirus infection can negatively affect vector reproduction, and that vertical transmission is a probable mechanism of overwintering of EHDV in North America.

Barely visible flower bud galls of Deutzia floribunda Nakai (Hydrangeaceae) were found in Kumamoto Prefecture, Kyushu, Japan. Morphological and molecular analyses indicated that the gall inducer is Pseudasphondylia tominagai Elsayed & Tokuda, 2019 (Diptera, Cecidomyiidae), previously known only to form galls on Eleutherococcus spinosus (L.f.) S.Y.Hu (Araliaceae) in Honshu, Japan. This study marks the first report of a species of Pseudasphondylia Monzen, 1955 on Hydrangeaceae, having host plants from different families and establishing D. floribunda as a new host record for P. tominagai.

Oropouche virus (OROV) is a vector-borne zoonotic virus that causes febrile illness in humans. Biting midges of the Culicoides genus are the primary vectors during human outbreaks. The 2022–2024 OROV outbreak has seen an increase in incidence, geographic expansion, and the emergence of previously undocumented symptoms. To better understand the basis of increased disease incidence, infection of the outbreak virus (OROV 240023 ) was compared to a historical virus strain (rOROV BeAn19991 ) in Culicoides sonorensis , a midge species that has demonstrated historical competence. Higher levels of infection, dissemination, and transmission potential were observed in C. sonorensis infected with the outbreak strain compared to the historical strain, although infectious titers did not differ between the two viruses. OROV 240023 was also detected in saliva at earlier time points than rOROV BeAn19991 , indicating a shorter extrinsic incubation period of < 5 days compared to 7–14 days for rOROV BeAn19991 . Taken together, our results demonstrate increased transmission potential of the outbreak strain in C. sonorensis midges, raising concern about the risk of spread within the United States following potential introduction. However, further studies are needed to evaluate the current strain in Culicoides species occurring within its outbreak range, including Culicoides paraensis , the confirmed South American vector of OROV.

Species distributions are dependent on interactions with abiotic and biotic factors in the environment. Abiotic factors like temperature, moisture, and soil nutrients, along with biotic interactions within and between species, can all have strong influences on spatial distributions of plants and animals. Terrestrial Antarctic habitats are relatively simple and thus good systems to study ecological factors that drive species distributions and abundance. However, these environments are also sensitive to perturbation, and thus understanding the ecological drivers of species distribution is critical for predicting responses to environmental change. The Antarctic midge, Belgica antarctica, is the only endemic insect on the continent and has a patchy distribution along the Antarctic Peninsula. While its life history and physiology are well studied, factors that underlie variation in population density within its range are unknown. Previous work on Antarctic microfauna indicates that distribution over broad scales is primarily regulated by soil moisture, nitrogen content, and the presence of suitable plant life, but whether these patterns are true over smaller spatial scales has not been investigated. Here we sampled midges across five islands on the Antarctic Peninsula and tested a series of hypotheses to determine the relative influences of abiotic and biotic factors on midge abundance. While historical literature suggests that Antarctic organisms are limited by the abiotic environment, our best-supported hypothesis indicated that abundance is predicted by a combination of abiotic and biotic conditions. Our results are consistent with a growing body of literature that biotic interactions are more important in Antarctic ecosystems than historically appreciated.

This study investigated frog‐biting dipteran species using newly designed frog‐calling traps in Taiwan. The trap effectively collected specimens from both families, Culicidae and Corethrellidae, demonstrating its utility. Host preference analysis revealed that Odorrana swinhoana (Boulenger, 1903) and Kurixalus eiffingeri (Boettger, 1895) were most frequently associated with collected specimens of Corethrellidae. Additionally, the corethrellids were predominantly attracted to a sound frequency around 2200 to 2700 Hz. Then, DNA barcoding was also conducted on the four collected species of Culicidae: Armigeres subalbatus (Coquillett, 1898), Uranotaenia nivipleura Leicester, 1908, Ur . macferlanei Edwards, 1914, and Mimomyia luzonensis (Ludlow, 1905), and the mitochondrial genome of Corethrella nippon Miyagi 1980 was first sequenced and annotated. Mitogenome‐based phylogenetic analysis confirmed that C. nippon formed a clade with Corethrella condita Borkent, 2008. In our analysis, family Corethrellidae clustered with Culicidae; however, the inter‐family phylogenetic relationships within Culicoidea appeared paraphyletic, particularly concerning family Chaoboridae. Future studies should explore a greater variety of frog species across more diverse regions and use other genomic datasets beyond the mitogenome to infer a more robust deep topology at the superfamily level and further broaden our understanding of host preference.

Background Biting midges in the genus Culicoides (Diptera: Ceratopogonidae) transmit bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV) to ruminants, thus exerting a significant economic impact on animal agriculture worldwide. However, very little is known about the larval habitat characteristics of Culicoides species associated with BTV/EHDV transmission, particularly in southeastern USA, limiting the establishment of effective midge control strategies. In this study, we examined the habitat associations of Culicoides species abundant on a commercial cervid farm in Florida, USA and quantified several environmental variables of their habitat to identify the key variables associated with midge abundance. Methods Mud/substrate samples from three potential larval habitats on the farm (edges of streams, puddles and seepages) were brought to the laboratory and incubated for adult emergence, and the percentage organic matter, macronutrients, micronutrients, pH, electrical conductivity, moisture and microbial concentrations of the substrate were quantified. Results Strong habitat associations were observed for Culicoides haematopotus (Malloch) (stream edge), Culicoides stellifer (Coquillett) (puddles) and Culicoides loisae (Jamnback) (stream edge), the most commonly emerging midge species from the samples. Suspected vector species of BTV/EHDV on the property, C. stellifer and Culicoides venustus (Hoffman), emerged mainly from habitats with moderate-high levels of pollution (edges of puddles and seepages) as indicated by the relatively higher concentrations/levels of organic matter, nutrients and other environmental variables in these samples. The emergence of C. insignis was too low to form any meaningful conclusions. For each Culicoides species, only weak positive or negative associations were detected between midge abundance and the various environmental variables quantified. Conclusions Habitat associations of Culicoides species abundant on a local cervid/animal farm vary, most likely as a function of certain biotic/abiotic characteristics of the habitat. Further studies across a larger spatial and temporal scale will be needed to experimentally evaluate/identify the key factors more strongly associated with the abundance of target Culicoides species. This information, in the long term, can be potentially exploited to render local habitats unsuitable for midge oviposition/larval development.