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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.

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.

The species Forcipomyia (Trichohelea) opilionivora (Lane, 1947) (Diptera, Ceratopogonidae) was rediscovered in the state of Rio de Janeiro, Brazil, 75 years after its original description. This study aimed to investigate the ectoparasitic relationship between F. (T.) opilionivora and its host and document its occurrence, which was recorded serendipitously during the collection of Opiliones (harvestmen) specimens in Parque Nacional do Itatiaia in the state of Rio de Janeiro, Brazil. After the collection, both specimens were subjected to photographic documentation and preserved for further examination. This study presents the first photographic record of the ectoparasitic relationship between F. (T.) opilionivora and a harvestman, shedding light on a poorly studied interaction. The host specimen was identified as a female of Holcobunus nigripalpis Roewer, 1910 (Opiliones, Sclerosomatidae), a gagrelline commonlyfound in the Atlantic Forest of southeastern Brazil. Previous records of ectoparasitism between Ceratopogonidae and harvestmen were limited to the family level (Sclerosomatidae species, specifically Gagrellinae or Leiobuninae). However, no specific host species had been identified until now. The rediscovery of F. (T.) opilionivora, along with the identification of its host, addresses a significant gap in our knowledge regarding the biology and distribution of this species and provides valuable insights into the intricate relationships between biting midges and arachnids. This study emphasizes the need for further investigations into the biology of these ectoparasitic species and underscores the importance of documenting and studying lesser-known interactions within ecosystems.

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.

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.

Culicoides insignis Lutz is incriminated as a vector of bluetongue virus (BTV) to ruminants in America. In South America, almost all countries have serological evidence of BTV infections, but only four outbreaks of the disease have been reported. Although clinical diseases have never been cited in Argentina, viral activity has been detected in cattle. In this study, we developed a potential distribution map of Culicoides insignis populations in northwestern Argentina using Maximum Entropy Modeling (Maxent). For the analyses, information regarding both data of specimen collections between 2003 and 2013, and climatic and environmental variables was used. Variables selection was based on the ecological relevance in relation to Culicoides spp. biology and distribution in the area. The best Maxent model according to the Jackknife test included 53 C. insignis presence records and precipitation of the warmest quarter, altitude, and precipitation of the wettest month. Accuracy was evaluated by the area under the curve (AUC = 0.97). These results provide an important analytical resource of high potential for both the development of suitable control strategies and the assessment of disease transmission risk in the region.

Insecticide formulations with safer environmental profiles and limited off-target effects are desirable to manage medical and veterinary pests. Silver nanoparticles are insecticidal against mosquitos, nonbiting midges, and other insects. The biting midge, Culicoides sonorensis Wirth and Jones, is a vector of agriculturally important pathogens in much of the United States, and this study aimed to examine the insecticidal properties of silver nanoparticles in larvae of this species. Mortality of third-instar larvae was assessed daily for 7 days after exposure to concentrations of silver nanoparticles, sorghum polymer particles, and hybrid silver–sorghum polymer particles. Both silver nanoparticles and silver–sorghum polymer particles were insecticidal, but sorghum polymer particles alone did not significantly contribute to larval mortality. Concentrations of 100 mg/liter of silver nanoparticles achieved >50% mortality at day 7, and 200 mg/liter treatments achieved >75% larval mortality within 24 h. The antimicrobial properties of silver nanoparticles were also examined, and culturable bacteria were recovered from larval-rearing media at 200 mg/liter but not at 400 mg/liter of silver nanoparticles. These data suggest that C. sonorensis larval mortality is primarily caused by silver nanoparticle toxicity and not by the reduction of bacteria (i.e., a larval food source). This work describes the first use of silver nanoparticles in C. sonorensis and shows the potential insecticide applications of these nanoparticles against this agricultural pest. The grain-polymer particles also successfully carried insecticidal silver nanoparticles, and their utility in loading diverse compounds could be a novel toxin delivery system for biting midges and similar pests. Graphical Abstract

Culicoides biting midges, small hematophagous dipterans, are the demonstrated or putative vectors of multiple arboviruses of veterinary and public health importance. Despite its relevance in disease spread, the ceratopogonid genus Culicoides is still a largely neglected group of species, predominantly because the major human-affecting arboviruses are considered to be transmitted by mosquitoes. However, when a pathogen is detected in a certain vector species, a thorough search for further vectors often remains undone and, therefore, the relevant vector species may remain unknown. Furthermore, for many hematophagous arthropods, true vector competence is often merely suspected and not experimentally proven. Therefore, we aim to illuminate the general impact of Culicoides biting midges and to summarize the knowledge about biting midge-borne disease agents using the order Bunyavirales, the largest and most diverse group of RNA viruses, as an example. When considering only viruses evidentially transmitted by Culicoides midges, the Simbu serogroup (genus Orthobunyavirus) is presumably the most important group within the virus order. Its members are of great veterinary importance, as a variety of simbuviruses, e.g., the species Akabane orthobunyavirus or Schmallenberg orthobunyavirus, induces severe congenital infections in pregnant animals. The major zoonotic representative of this serogroup occurs in South and Central America and causes the so-called Oropouche fever, an acute febrile illness in humans.

Biting midges of the genus Leptoconops (Diptera: Ceratopogonidae) are neglected yet aggressive diurnal biting pests. This study investigates the species diversity, abundance, biting behaviour, trap efficiency, and public health relevance of Leptoconops across ten tourist and potential tourist beaches in the Andaman and Nicobar archipelago, India, during two peak tourist seasons. Three species were identified: Leptoconops (Styloconops) spinosifrons , first report from the archipelago; Leptoconops (Leptoconops) leptorhynchus , newly recorded from India and redescribed; Leptoconops (Leptoconops) paruii sp. nov., a proposed new species. Species identification was supported by morphological characters and a COI-based molecular phylogeny along with species delimitation analysis using ABGD, ASAP and mPTP. Biting activity studies revealed unimodal and bimodal biting patterns for L. paruii and L. spinosifrons , respectively. Inter-island distribution and abundance patterns revealed L. spinosifrons (most abundant) was restricted to the Andaman group, L. paruii occurred in both the Andaman group and the Great Nicobar, with L. leptorhynchus being the rarest. Among diurnal capture methods, HLC proved most effective for capturing host-seeking females, while sweep netting captured both sexes. Nocturnally, LED light sheets outperformed UV-based CDC and UT. Furthermore, the case report of Leptoconops bite-induced dermatitis highlights the potential clinical relevance of these bites for travellers and local population.

Bacteria are part of the insect gut system and influence many physiological traits of their host. Gut bacteria may even reduce or block the transmission of arboviruses in several species of arthropod vectors. Culicoides biting midges are important arboviral vectors of several livestock and wildlife diseases, yet limited information is available on their gut bacterial communities. Addressing this gap will help inform how these communities can be manipulated and ultimately used as novel tools to control pathogens. To assess how bacterial communities change during the life stages of lab-reared C. nubeculosus and C. sonorensis, endosymbiotic bacteria were identified using Illumina sequencing of 16S rRNA and taxonomically characterised. Analyses were conducted to determine how gut bacterial communities in adults are influenced by species identity and geographic distance among biting midge populations. Communities of the two lab-reared Culicoides species significantly changed after pupation and with maturation into 6-day-old adults. Pseudomonas, Burkholderiaceae and Leucobacter bacteria were part of a core community that was trans-stadially transmitted and found throughout their life cycle. Among field-collected biting midges, the bacterial communities were unique for almost each species. Cardinium, Rickettsia and Wolbachia were some of the most abundant bacteria in midges collected from wetlands. Only Pseudomonas was present in high relative abundance in all field-collected species. In this study, species identity, as well as geographic distance, influenced the gut bacterial communities and may partly explain known inter- and intra-species variability in vector competence. Additionally, stably associated bacterial species could be candidates for paratransgenic strategies to control vector-borne pathogens.