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BACKGROUND: As a primary vector of bluetongue virus (BTV) in the US, seasonal abundance and diel flight activity of Culicoides sonorensis has been documented, but few studies have examined how time of host-seeking activity is impacted by environmental factors. This knowledge is essential for interpreting surveillance data and modeling pathogen transmission risk. METHODS: The diel host-seeking activity of C. sonorensis was studied on a California dairy over 3 years using a time-segregated trap baited with CO2. The relationship between environmental variables and diel host-seeking activity (start, peak, and duration of activity) of C. sonorensis was evaluated using multiple linear regression. Fishers exact test and paired-sample z-test were used to evaluate the seasonal difference and parity difference on diel host-seeking activity. RESULTS: Host-seeking by C. sonorensis began and reached an activity peak before sunset at a higher frequency during colder months relative to warmer months. The time that host-seeking activity occurred was associated low and high daily temperature as well as wind speed at sunset. Colder temperatures and a greater diurnal temperature range were associated with an earlier peak in host-seeking. Higher wind speeds at sunset were associated with a delayed peak in host-seeking and a shortened duration of host-seeking. Parous midges reached peak host-seeking activity slightly later than nulliparous midges, possibly because of the need for oviposition by gravid females before returning to host-seeking. CONCLUSIONS: This study demonstrates that during colder months C. sonorensis initiates host-seeking and reaches peak host-seeking activity earlier relative to sunset, often even before sunset, compared to warmer months. Therefore, the commonly used UV light-baited traps are ineffective for midge surveillance before sunset. Based on this study, surveillance methods that do not rely on light trapping would provide a more accurate estimate of host-biting risk across seasons. The association of environmental factors to host-seeking shown in this study can be used to improve modeling or prediction of host-seeking activity. This study identified diurnal temperature range as associated with host-seeking activity, suggesting that Culicoides may respond to a rapidly decreasing temperature by shifting to an earlier host-seeking time, though this association needs further study.

Culicoides are vectors that can transmit many different pathogens to mammals — including humans, and domestic and wild animals — and birds. In order to take preventive measures against any vector-borne disease, it is important to gather information on both the host and vector species. Culicoides species are mainly mammalophilic, ornithophilic or ornithophilic/mammalophilic, but females have also been found to occasionally feed on engorged insects. A recent systematic review based on three groups of key words investigated Culicoides on farms, and asserted that 92 species (including four not present species) have been reported among cattle in mainland France and Corsica. We have re-evaluated the presence of Culicoides species in cattle in France using the same data of the review. Our data show that only 18 species are reported among cattle. Furthermore, our research used molecular and indirect investigations to analyse Culicoides species that had been feeding on cattle. Our results demonstrate that 45 species feed on cattle out of 92 species present in France. The paper discusses the relevance of data in the literature when investigating hosts of Culicoides species. [ABSTRACT FROM AUTHOR]

Culicoides, among the tiniest and most abundant hematophagous insects globally, serve as vectors for a variety of pathogens such as viruses, bacteria, parasites, protozoa, and nematodes. This study aimed to identify Culicoides species and assess their spatial distribution and seasonal occurrence in selected districts of the Central, South, and West Gondar zones, Northwest Ethiopia. A cross-sectional study was conducted between January to July 2023. A total of 44 UV light- onderstepoort traps were deployed in the study districts near specific areas. The traps were operational from dusk (6:00 PM) until dawn (6:00 AM) and were suspended at a height of 1.5 to 2 m above the ground. Poisson regression was used to assess associations, the Shannon diversity index to measure diversity, and QGIS 3.22.6 to create maps. In this study, 8,857 Culicoides were captured across the 44 trapping sites. Of the total flies captured flies, 8,838 were identified as belonging to 12 distinct species, while the classification of the remaining 19 flies remained unclear. Notably, C. kingi (54.01%) was the most prevalent species, followed by C. imicola (44.55%). The abundance of Culicoides observed from January to late April (3505) was significantly lower compared to the wet season (5355), with a marked increase in the capture of C. kingi (2499) from May to late July. A statistically significant association (p < 0.05) was observed between the occurrence of Culicoides and factors such as district, sampling point, and season. Spatial analysis revealed that C. kingi had a broader range of suitability than other Culicoides species, with high suitability observed in East Dembia. The diversity index analysis indicated that Culicoides species diversity was higher in samples from animal pens (H = 0.73) and during the wet season (H = 0.75). Additionally, this study documented the presence of eight Culicoides species namely C. corsicus, C. kibunensis, C. reioxi, C. kiouxi, C. saharienines, C. desertorum, C. reithi, and C. festivipennis, which have not been previously documented in Ethiopia. In conclusion, the study highlighted that the occurrence of Culicoides species was higher in East Dembia, with moderate presence in Wegera and West Armacho. Further research is needed to assess the impact of various Culicoides species on animal and human health, as well as their economic implications, and to develop corresponding control strategies based on these findings. [ABSTRACT FROM AUTHOR]

Background: Clinical cases of leishmaniasis caused by Leishmania (Mundinia) parasites have been increasingly reported in Southeast Asia, particularly Thailand. Recent evidence has shown that Leishmania (Mundinia) parasites successfully developed into infective metacyclic promastigotes in Culicoides biting midges, strongly supporting their putative role in disease transmission. However, Culicoides diversity, host preference, and Leishmania prevalence in endemic areas remain largely unknown. Methods: We investigated the seasonal dynamics, infection prevalence, and blood meal identification of Culicoides collected from the emerging focus of visceral leishmaniasis in Lampang Province, Northern Thailand, during 2021–2023. Midge samples were molecularly screened for Leishmania using SSU rRNA-qPCR and ITS1-PCR, followed by Sanger plasmid sequencing, and parasite haplotype diversity was analyzed. Host blood meal origins were comparatively identified using host-specific Cytb-PCRs and a nanopore-based metabarcoding approach. Results: A total of 501 parous and gravid females and 46 blood-engorged ones belonging to at least 17 species of five subgenera (Remmia, Trithecoides, Avaritia, Hoffmania, and Meijerehelea) and two species groups (Shortti and Calvipalpis) were collected with temporal differences in abundance. Leishmania was detected by SSU rRNA-qPCR in 31 samples of at least 11 midge species, consisting of Culicoides oxystoma, C. guttifer, C. orientalis, C. mahasarakhamense, C (Trithecoides) spp., C. innoxius, C. shortti, C. arakawae, C. sumatrae, C. actoni, and C. fulvus, with the overall infection prevalence of 5.7%. The latter six species represent the new records as putative leishmaniasis vectors in Northern Thailand. The ITS1-PCR and plasmid sequencing revealed that Leishmania martiniquensis was predominantly identified in all qPCR-positive species, whereas L. orientalis was identified only in three C. oxystoma samples. The most dominant haplotype of L. martiniquensis in Thailand was genetically intermixed with those from other geographical regions, confirming its globalization. Neutrality test statistics were also significantly negative on regional and country-wide scales, suggesting rapid population expansion or selective sweeps. Nanopore-based blood meal analysis revealed that most Culicoides species are mammalophilic, with peridomestic and wild mammals (cow, pig, deer, and goat-like species) and humans as hosts, while C. guttifer and C. mahasarakhamense fed preferentially on chickens. Conclusions: This study revealed seasonal dynamics and sympatric circulation of L. martiniquensis and L. orientalis in different species of Culicoides. Evidence of human blood feeding was also demonstrated, implicating Culicoides as putative vectors of human leishmaniasis in endemic areas. Further research is therefore urgently needed to develop vector control strategies and assess the infection status of their reservoir hosts to effectively minimize disease transmission. [ABSTRACT FROM AUTHOR]

Simple Summary: The emergence of midge-borne diseases such as lumpy skin disease in Thailand highlights the necessity of understanding the seasonal abundance and spatial distribution of Culicoides biting midges. Population dynamics, species composition, and host preferences of Culicoides species were studied in Kanchanaburi, Thailand. Nine UV light traps were deployed to collect midges in nine animal sheds over the course of a year, revealing abundant species such as C. oxystoma Kieffer and C. peregrinus Kieffer, C. arakawae Arakawa, and C. imicola Kieffer. These species demonstrated preferences for different farm animals and peaked in trap captures from the late rainy season to early winter. Trap catches were positively influenced by relative humidity and temperature, while they exhibited a negative impact from wind speed. This study provides important data for disease management and prevention, emphasizing the impact of these vectors on both animal and human health. Overall, this research underscores the importance of ongoing studies into Culicoides behavior, ecology, and disease transmission potential to inform effective control strategies. Culicoides biting midges were collected using UV light traps from nine livestock farms in Kanchanaburi Province, Thailand. Collections were made one night per month from June 2020 to May 2021 to determine the seasonal changes and spatial distribution of the Culicoides assemblage. The influence of four environmental factors (temperature, rainfall, humidity, and wind speed), types of animals in each shed (cattle, pigs, and avians), and neighboring animals (those in the other sheds and their proximity) were assessed. A subsample of 130,670 out of a total of 224,153 specimens collected was identified and counted. The collections were predominantly female (76.9%), though males were also commonly collected (23.1%). The dominant species included C. peregrinus (97,098 individuals; 43.0%), C. oxystoma (55,579; 24.6%), C. arakawae (45,996; 20.4%), and C. imicola (15,703; 7.0%), while other species accounted for the remaining 9777 individuals (4.4%). Trap captures were strongly influenced by seasons and animal sheds. Cattle exhibited the greatest effect on the community, likely due to their large biomass. Humidity and temperature had a positive effect on trap captures, whereas wind speed exhibited a negative effect. Cattle positively influenced all major species, except for C. arakawae, which showed a positive association with avians. A "neighboring effect" was also observed. Additionally, the lowest dispersal ability of C. arakawae was suggested. [ABSTRACT FROM AUTHOR]

Culicoides biting midges are small dipterous insects known as biological vectors of arboviruses, protozoa, and filaria parasites worldwide. Many studies on Culicoides focus on trapping them at ground level, without considering the best trap heights for different vector species. This implies that we might overlook insects positioned higher in the canopy. From June to August, we used UV traps to catch Culicoides biting midges at three different heights in three temperate mature forest areas in east Lithuania, Baltic region of Europe. We conducted this study to test the differences in midge numbers, male and female proportions, and female parity at each height. We caught the majority of biting midges (80.6%) at the mid-canopy and high-canopy. A higher number of female Culicoides midges than males was caught, with the proportion of males varying based on height and reaching its lowest point at ground level. No significant difference between the proportion of nulliparous and parous females caught at different height was detected. Culicoides pictipennis and C. festivipennis were the most common species of biting midge we found. They were found in the mid-canopy (86.8%) and the high-canopy (50.0%), respectively. Culicoides kibunensis was next, found at ground level (66.2%), and C. punctatus was found at the high canopy strata (63.0%). Each species' abundance was seasonal dependent. Information on the vertical distribution of vector species in the temperate forest ecosystem is an important step in understanding patterns of vector borne disease transmission in wildlife. [ABSTRACT FROM AUTHOR]

Bluetongue (BT) is a Culicoides midge-borne hemorrhagic disease affecting cervids and ruminant livestock species, resulting in significant economic losses from animal production and trade restrictions. Experimental animal infections using the α/β interferon receptor knockout IFNAR mouse model and susceptible target species are critical for understanding viral pathogenesis, virulence, and evaluating vaccines. However, conducting experimental vector-borne transmission studies with the vector itself are logistically difficult and experimentally problematic. Therefore, experimental infections are induced by hypodermic injection with virus typically derived from baby hamster kidney (BHK) cells. Unfortunately, for many U.S. BTV serotypes, it is difficult to replicate the severity of the disease seen in natural, midge-transmitted infections by injecting BHK-derived virus into target host animals. Using the IFNAR BTV murine model, we compared the virulence of traditional BHK cell-derived BTV-17 with C. sonorensis midge (W8) cell-derived BTV-17 to determine whether using cells of the transmission vector would provide an in vitro virulence aspect of vector-transmitted virus. At both low and high doses, mice inoculated with W8-BTV-17 had an earlier onset of viremia, earlier onset and peak of clinical signs, and significantly higher mortality compared to mice inoculated with BHK-BTV-17. Our results suggest using a Culicoides W8 cell-derived inoculum may provide an in vitro vector-enhanced infection to more closely represent disease levels seen in natural midge-transmitted infections while avoiding the logistical and experimental complexity of working with live midges. [ABSTRACT FROM AUTHOR]

Vesicular stomatitis (VS) is a viral disease that affects horses, cattle, and swine that is transmitted by direct contact and hematophagous insects. In 2023, a multi-state outbreak of vesicular stomatitis New Jersey virus (VSNJV) occurred in California, Nevada, and Texas, infecting horses, cattle, and rhinoceros. To identify possible insect vectors, we conducted insect surveillance at various locations in San Diego County, CA, including at a wildlife park. CO2 baited traps set from mid-May to mid-August 2023 collected 2357 Culicoides biting midges and 1215 Simulium black flies, which are insect genera implicated in VSNJV transmission. Insects were pooled by species, location, and date, then tested for viral RNA. Nine RNA-positive pools of Culicoides spp. and sixteen RNA-positive pools of Simulium spp were detected. Infectious virus was detected by cytopathic effect in 96% of the RNA-positive pools. This is the first report of VSNJV in wild-caught C. bergi, C. freeborni, C. occidentalis, S. argus, S. hippovorum, and S. tescorum. The vector competency of these species for VSNJV has yet to be determined but warrants examination. Active vector surveillance and testing during disease outbreaks increases our understanding of the ecology and epidemiology of VS and informs vector control efforts. [ABSTRACT FROM AUTHOR]

Introduction: Culicoides Latreille biting midges are vectors of high concern as they can transmit serious veterinary diseases such as bluetongue virus or epizootic haemorrhagic disease virus, among others. Little is known about these vectors in Galicia, so a comprehensive literature review and an intensive monitoring were carried out in the region. Material and methods: The Autonomous Community of Galicia was sampled through three different vector surveillance projects between 2004 and 2023. A total of 239 sampling points were deployed alongside the Galician territory. In addition, a literature review of Culicoides in Galicia related content was made by consulting several digital repositories. Results: A total of 33 species of Culicoides belonging to 8 subgenera were identified. Among them, 15 are considered or suspected to be potential vectors of several pathogens of medical and/or veterinary interest. In addition, 20 of them are reported for the first time in the region. Updated distribution maps of the Culicoides biting midges of Galicia were provided, including several notes regarding their ecology and relevance for both public health and animal welfare. Conclusions: The present work is one of the most complete works made at regional level in Spain to date. As Galicia's economy relies heavily on livestock farming, this work will provide a solid baseline in order to develop new research lines in the future regarding prevention to vector‐borne diseases. [ABSTRACT FROM AUTHOR]

Culicoides are responsible for a number of diseases transmitted to the animals causing severe health conditions and great economic losses. The outbreak of bluetongue virus in Albania in 2014 and lumpy skin disease in 2016 strongly indicated the establishment of a sentinel herd surveillance in the country ruminant farms. Considering all evidence for Albania, investigation of species composition and abundance of Culicoides on livestock farms was performed in 10 sentinel sites. Ultraviolet-light traps (OVI-traps) were placed in selected sites for collection of Culicoides starting from April 15 to July 30, 2021. A total of 37,111 Culicoides midges from 18 different species were collected during the investigation. The most abundant species identified were C. newsteadi and C. obsoletus. Moreover, three species were described for the first time in Albania (C. collessi, C. chiopterus and C. immaculatus). Presence and abundance of the collected Culicoides spp. are associated with potential outbreak of diseases (i.e. bluetongue, ephemeral fever) linked with their transmission from Culicoides. [ABSTRACT FROM AUTHOR]

Abstract Culicoides, among the tiniest and most abundant hematophagous insects globally, serve as vectors for a variety of pathogens such as viruses, bacteria, parasites, protozoa, and nematodes. This study aimed to identify Culicoides species and assess their spatial distribution and seasonal occurrence in selected districts of the Central, South, and West Gondar zones, Northwest Ethiopia. A cross-sectional study was conducted between January to July 2023. A total of 44 UV light- onderstepoort traps were deployed in the study districts near specific areas. The traps were operational from dusk (6:00 PM) until dawn (6:00 AM) and were suspended at a height of 1.5 to 2 m above the ground. Poisson regression was used to assess associations, the Shannon diversity index to measure diversity, and QGIS 3.22.6 to create maps. In this study, 8,857 Culicoides were captured across the 44 trapping sites. Of the total flies captured flies, 8,838 were identified as belonging to 12 distinct species, while the classification of the remaining 19 flies remained unclear. Notably, C. kingi (54.01%) was the most prevalent species, followed by C. imicola (44.55%). The abundance of Culicoides observed from January to late April (3505) was significantly lower compared to the wet season (5355), with a marked increase in the capture of C. kingi (2499) from May to late July. A statistically significant association (p

Culicoides are responsible for a number of diseases transmitted to the animals causing severe health conditions and great economic losses. The outbreak of bluetongue virus in Albania in 2014 and lumpy skin disease in 2016 strongly indicated the establishment of a sentinel herd surveillance in the country ruminant farms. Considering all evidence for Albania, investigation of species composition and abundance of Culicoides on livestock farms was performed in 10 sentinel sites. Ultraviolet-light traps (OVI-traps) were placed in selected sites for collection of Culicoides starting from April 15 to July 30, 2021. A total of 37,111 Culicoides midges from 18 different species were collected during the investigation. The most abundant species identified were C. newsteadi and C. obsoletus. More-over, three species were described for the first time in Albania (C. collessi, C. chiopterus and C. im-maculatus). Presence and abundance of the collected Culicoides spp. are associated with potential outbreak of diseases (i.e. bluetongue, ephemeral fever) linked with their transmission from Culi-coides.

The Leucocytozoon parasites are one of the important protozoa species affecting domestic poultry and wild birds. Though there are frequent reports about the incidence of Leucocytozoon in wild birds, the information regarding the occurrence of commercial poultry is underestimated. From October 2015 to August 2022, ailing and dead birds from 39 commercial layer flocks were submitted to the Poultry Disease Diagnosis and Surveillance Laboratory, Namakkal, with a history of brief illness, followed by mortality with oozing of blood from oro-nasal orifice. The presence of Leucocytozoon caulleryi was confirmed in all the flocks by laboratory examination and PCR. All the Leucocytozoon-positive cases were reported after heavy rainfall and the affected farms were inhabited with Culicoides flies. The infected birds were dull and depressed, exhibited labored breathing, and blood-tinted ropy mucous from the oro-nasal cavity. Necropsy examination revealed pale-anaemic comb and wattles, massive haemorrhagic clot over the surface of the kidney, peritoneum, and trachea, and scattering of greyish-white/reddish military megaloschizonts throughout the serosa of viscera and skeletal muscles. Histopathological examination revealed many spherical cysts or megaloschizonts in the intestine, pancreas, gizzard, spleen, liver, kidney, heart, and breast muscles. The cysts were deeply seated within the above organ's parenchyma or embedded in the serosa. There was extensive connective tissue proliferation with mononuclear cell infiltration around the cyst. Leucocytozoon-specific DNA was detected in all the flocks by PCR. Phylogenetic analysis of the cytochrome b gene of L. caulleryi reported from various parts of the world revealed that L. caulleryi is grouped into two major clusters (Cluster I & II). Further, Indian L. caulleryi sequences fall under sub-cluster I and II along with L. caulleryi reported from Taiwan, Egypt, China, Myanmar, Japan, South Korea, and Thailand. [ABSTRACT FROM AUTHOR]

Abstract Background Culicoides biting midges exhibit a global spatial distribution and are the main vectors of several viruses of veterinary importance, including bluetongue (BT) and African horse sickness (AHS). Many environmental and anthropological factors contribute to their ability to live in a variety of habitats, which have the potential to change over the years as the climate changes. Therefore, as new habitats emerge, the risk for new introductions of these diseases of interest to occur increases. The aim of this study was to model distributions for two primary vectors for BT and AHS (Culicoides imicola and Culicoides bolitinos) using random forest (RF) machine learning and explore the relative importance of environmental and anthropological factors in a region of South Africa with frequent AHS and BT outbreaks. Methods Culicoides capture data were collected between 1996 and 2022 across 171 different capture locations in the Western Cape. Predictor variables included climate-related variables (temperature, precipitation, humidity), environment-related variables (normalised difference vegetation index—NDVI, soil moisture) and farm-related variables (livestock densities). Random forest (RF) models were developed to explore the spatial distributions of C. imicola, C. bolitinos and a merged species map, where both competent vectors were combined. The maps were then compared to interpolation maps using the same capture data as well as historical locations of BT and AHS outbreaks. Results Overall, the RF models performed well with 75.02%, 61.6% and 74.01% variance explained for C. imicola, C. bolitinos and merged species models respectively. Cattle density was the most important predictor for C. imicola and water vapour pressure the most important for C. bolitinos. Compared to interpolation maps, the RF models had higher predictive power throughout most of the year when species were modelled individually; however, when merged, the interpolation maps performed better in all seasons except winter. Finally, midge densities did not show any conclusive correlation with BT or AHS outbreaks. Conclusion This study yielded novel insight into the spatial abundance and drivers of abundance of competent vectors of BT and AHS. It also provided valuable data to inform mathematical models exploring disease outbreaks so that Culicoides-transmitted diseases in South Africa can be further analysed. Graphical Abstract

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.

Background: Culicoides biting midges exhibit a global spatial distribution and are the main vectors of several viruses of veterinary importance, including bluetongue (BT) and African horse sickness (AHS). Many environmental and anthropological factors contribute to their ability to live in a variety of habitats, which have the potential to change over the years as the climate changes. Therefore, as new habitats emerge, the risk for new introductions of these diseases of interest to occur increases. The aim of this study was to model distributions for two primary vectors for BT and AHS (Culicoides imicola and Culicoides bolitinos) using random forest (RF) machine learning and explore the relative importance of environmental and anthropological factors in a region of South Africa with frequent AHS and BT outbreaks. Methods: Culicoides capture data were collected between 1996 and 2022 across 171 different capture locations in the Western Cape. Predictor variables included climate-related variables (temperature, precipitation, humidity), environment-related variables (normalised difference vegetation index—NDVI, soil moisture) and farm-related variables (livestock densities). Random forest (RF) models were developed to explore the spatial distributions of C. imicola, C. bolitinos and a merged species map, where both competent vectors were combined. The maps were then compared to interpolation maps using the same capture data as well as historical locations of BT and AHS outbreaks. Results: Overall, the RF models performed well with 75.02%, 61.6% and 74.01% variance explained for C. imicola, C. bolitinos and merged species models respectively. Cattle density was the most important predictor for C. imicola and water vapour pressure the most important for C. bolitinos. Compared to interpolation maps, the RF models had higher predictive power throughout most of the year when species were modelled individually; however, when merged, the interpolation maps performed better in all seasons except winter. Finally, midge densities did not show any conclusive correlation with BT or AHS outbreaks. Conclusion: This study yielded novel insight into the spatial abundance and drivers of abundance of competent vectors of BT and AHS. It also provided valuable data to inform mathematical models exploring disease outbreaks so that Culicoides-transmitted diseases in South Africa can be further analysed. [ABSTRACT FROM AUTHOR]

Background: Culicoides midges have been well-studied in Spain, particularly over the last 20 years, mainly because of their role as vectors of arboviral diseases that affect livestock. Most studies on Culicoides are conducted using suction light traps in farmed environments, but studies employing alternative trapping techniques or focusing on natural habitats are scarce. Methods: In the present study, we analyze Culicoides captured in 2023 at 476 sites in western Andalusia (southern Spain) using carbon dioxide-baited Biogents (BG)-sentinel traps across different ecosystems. Results: We collected 3,084 Culicoides midges (3060 females and 24 males) belonging to 23 species, including the new species Culicoides grandifovea sp. nov. and the first record of Culicoides pseudolangeroni for Europe. Both species were described with morphological and molecular methods and detailed data on spatial distribution was also recorded. The new species showed close phylogenetic relations with sequences from an unidentified Culicoides from Morocco (92.6% similarity) and with Culicoides kurensis. Culicoides imicola was the most abundant species (17.4%), followed by Culicoides grandifovea sp. nov. (14.6%) and Culicoides kurensis (11.9%). Interestingly, Culicoides montanus was the only species of the obsoletus and pulicaris species complexes captured, representing the first record of this species in southern Spain. A total of 53 valid Culicoides species have been reported in the area, with 48 already reported in literature records and 5 more added in the present study. Information on the flight period for the most common Culicoides species is also provided. Conclusions: To the best of our knowledge, our study represents the most comprehensive effort ever done on nonfarmland habitats using carbon-dioxide baited suction traps for collecting Culicoides. Our data suggests that using carbon dioxide traps offers a completely different perspective on Culicoides communities compared with routinely used light traps, including the discovery of previously unrecorded species. [ABSTRACT FROM AUTHOR]

Most mathematical models that assess the vectorial capacity of disease-transmitting insects typically focus on the influence of climatic factors to predict variations across different times and locations, or examine the impact of vector control interventions to forecast their potential effectiveness. We combine features of existing models to develop a novel model for vectorial capacity that considers both climate and vector control. This model considers how vector control tools affect vectors at each stage of their feeding cycle, and incorporates host availability and preference. Applying this model to arboviruses of veterinary importance in Europe, we show that African horse sickness virus (AHSV) has a higher peak predicted vectorial capacity than bluetongue virus (BTV), Schmallenberg virus (SBV), and epizootic haemorrhagic disease virus (EHDV). However, AHSV has a shorter average infectious period due to high mortality; therefore, the overall basic reproduction number of AHSV is similar to BTV. A comparable relationship exists between SBV and EHDV, with both viruses showing similar basic reproduction numbers. Focusing on AHSV transmission in the UK, insecticide-treated stable netting is shown to significantly reduce vectorial capacity of Culicoides, even at low coverage levels. However, untreated stable netting is likely to have limited impact. Overall, this model can be used to consider both climate and vector control interventions either currently utilised or for potential use in an outbreak, and could help guide policy makers seeking to mitigate the impact of climate change on disease control. [ABSTRACT FROM AUTHOR]

In 2021, a pregnant Rottweiler dog living on a sheep farm was diagnosed with clinical bluetongue (BT) infection. This study reports on the investigation of this farm where bluetongue virus (BTV) infection was diagnosed in this atypical host species. Samples were collected during farm visits 14, 28, 60, and 89 days after the onset of clinical signs in the pregnant Rottweiler. Blood was collected from all farm dogs (n = 6) and tested for BTV genome using a reverse‐transcriptase quantitative PCR (RT‐qPCR) assay and BTV antibodies with the competitive ELISA (cELISA) and dogs positive by RT‐qPCR were further tested using virus neutralization (VN) serological testing. Blood was also collected from 16 sick sheep and tested using RT‐qPCR. Midges were trapped on the study farm using an Onderstepoort UV light trap placed above a sheep pen for 36 hr at the first farm (14 days) visit. Parous/gravid midges were tested by BTV RT‐qPCR in batches of up to 200 midges per species. Blood‐fed midges (n = 308) were tested using a PCR species probe (KAPA Multiplex Master Mix) to identify the host species on which the midge had fed. Three dogs (n = 3/6) had detectable BTV RNA with RT‐qPCR and high VN antibody titers to BTV. All RT‐qPCR‐positive dogs and one additional dog tested cELISA seropositive (n = 4/6). Bluetongue virus RNA was detected in 5/16 sheep tested. The most abundant midge species was Culicoides imicola (99.3%) and BTV was only detected in this species (n = 3/4 batches of 200 parous midges). Dog blood was not detected in any blood‐fed midges tested. The occurrence of natural BT viraemia in exposed dogs creates a potential risk of BTV entry into BT‐free countries through dog importation. It remains unclear whether BT viremia in dogs is capable of onward transmission. [ABSTRACT FROM AUTHOR]

The study aimed to investigate molecularly the presence of flea-borne viruses in infested small ruminants with fleas. It was carried out in Egypt's Northern West Coast (NWC) and South Sinai Governorate (SSG). Three specific primers were used targeting genes, ORF103 (for Capripoxvirus and Lumpy skin disease virus), NS3 (for Bluetongue virus), and Rdrp (for Coronavirus), followed by gene sequencing and phylogenetic analyses. The results revealed that 78.94% of sheep and 65.63% of goats were infested in the NWC area, whereas 49.76% of sheep and 77.8% of goats were infested in the SSG region. Sheep were preferable hosts for flea infestations (58.9%) to goats (41.1%) in the two studied areas. Sex and age of the animals had no effects on the infestation rate (p > 0.05). The season and site of infestation on animals were significantly different between the two areas (p < 0.05). Ctenocephalides felis predominated in NWC and Ctenocephalides canis in SSG, and males of both flea species were more prevalent than females. Molecular analysis of flea DNA revealed the presence of Capripoxvirus in all tested samples, while other viral infections were absent. Gene sequencing identified three isolates as sheeppox viruses, and one as goatpox virus. The findings suggest that Capripoxvirus is adapted to fleas and may be transmitted to animals through infestation. This underscores the need for ongoing surveillance of other pathogens in different regions of Egypt. [ABSTRACT FROM AUTHOR]

Biting midges (Culicoides) are vectors of many pathogens of medical and veterinary importance, but their viromes are poorly characterized compared to certain other hematophagous arthropods, e.g., mosquitoes and ticks. The goal of this study was to use metagenomics to identify viruses in Culicoides from Mexico. A total of 457 adult midges were collected in Chihuahua, northern Mexico, in 2020 and 2021, and all were identified as female Culicoides reevesi. The midges were sorted into five pools and homogenized. An aliquot of each homogenate was subjected to polyethylene glycol precipitation to enrich for virions, then total RNA was extracted and analyzed by unbiased high-throughput sequencing. We identified six novel viruses that are characteristic of viruses from five families (Nodaviridae, Partitiviridae, Solemoviridae, Tombusviridae, and Totiviridae) and one novel virus that is too divergent from all classified viruses to be assigned to an established family. The newly discovered viruses are phylogenetically distinct from their closest known relatives, and their minimal infection rates in female C. reevesi range from 0.22 to 1.09. No previously known viruses were detected, presumably because viral metagenomics had never before been used to study Culicoides from the Western Hemisphere. To conclude, we discovered multiple novel viruses in C. reevesi from Mexico, expanding our knowledge of arthropod viral diversity and evolution. [ABSTRACT FROM AUTHOR]

The article focuses on the spread and impact of bluetongue virus (BTV), specifically BTV serotype 3 (BTV-3), in European ruminants, highlighting its emergence in the Netherlands and subsequent spread to Belgium, the United Kingdom, and Germany by 2023, necessitating regulatory responses under EU guidelines to manage outbreaks.

Bluetongue virus (BTV, Sedoreoviridae: Orbivirus) causes an economically important disease, namely, bluetongue (BT), in domestic and wild ruminants worldwide. BTV is endemic to South India and has occurred with varying severity every year since the virus was first reported in 1963. BT can cause high morbidity and mortality to sheep flocks in this region, resulting in serious economic losses to subsistence farmers, with impacts on food security. The epidemiology of BTV in South India is complex, characterized by an unusually wide diversity of susceptible ruminant hosts, multiple vector species biting midges (Culicoides spp., Diptera: Ceratopogonidae), which have been implicated in the transmission of BTV and numerous co-circulating virus serotypes and strains. BT presence data (1997–2011) for South India were obtained from multiple sources to develop a presence/absence model for the disease. A non-linear discriminant analysis (NLDA) was carried out using temporal Fourier transformed variables that were remotely sensed as potential predictors of BT distribution. Predictive performance was then characterized using a range of different accuracy statistics (sensitivity, specificity, and Kappa). The top ten variables selected to explain BT distribution were primarily thermal metrics (land surface temperature, i.e., LST, and middle infrared, i.e., MIR) and a measure of plant photosynthetic activity (the Normalized Difference Vegetation Index, i.e., NDVI). A model that used pseudo-absence points, with three presence and absence clusters each, outperformed the model that used only the recorded absence points and showed high correspondence with past BTV outbreaks. The resulting risk maps may be suitable for informing disease managers concerned with vaccination, prevention, and control of BT in high-risk areas and for planning future state-wide vector and virus surveillance activities. [ABSTRACT FROM AUTHOR]

We performed whole genome sequencing (WGS) of 15 Palyam serogroup virus (PALV) strains isolated from cattle or Culicoides biting midges in Japan from 1984 to 2018. We found that the PALV strains consisted of Chuzan (Kasba) virus (CHUV), D'Aguilar virus (DAGV), Bunyip Creek virus, and another PALV, Marrakai virus (MARV). The Japanese MARV strains isolated in 1997 were closely related to Australian PALV strains isolated in 1968–1976 in genome segments 2 and 10, but they were most closely related to other Japanese PALV strains in the other genome segments. Our data suggest that the Japanese MARV strains were reassortant viruses between Asian and Australian PALVs. In addition to the WGS, we developed a real-time reverse-transcription polymerase chain reaction assay that can broadly detect PALV and specifically detect CHUV and DAGV, utilizing the data obtained by the WGS in this study. We detected the DAGV gene in bovine stillborn fetuses and congenitally abnormal calves in 2019 using the newly developed assay. To our knowledge, this is the first report of isolation of MARV outside of Australia and the first report of detection of PALV in bovine fetuses or calves with congenital abnormality outside of Africa. [ABSTRACT FROM AUTHOR]

The epizootic hemorrhagic disease virus (EHDV) is a novel emerging threat for the European livestock sector. First detected in Sardinia and southern Spain at the end of 2022, this transboundary disease emerged in France in September 2023 despite restrictions on animal movement and enhanced surveillance protocols. Although virus spread is believed to be mediated by the dispersal of Culicoides vectors by the wind, prediction is difficult due to the large number of meteorological parameters that must be considered. Using simulations of atmospheric trajectories, we developed a model to investigate the long‐distance dispersal risk zone of Culicoides in Europe, starting from different source zones. Our model predicted with good sensitivity the newly EHDV‐infected areas in France over a period of 5 weeks after its first introduction in the country. Prospectively, we predicted that the midge dispersal zone of early 2024 could expand toward most of the western half of France and could sporadically reach new countries under favorable spring conditions. The wind dispersal risk maps provided are intended to support better preparedness and response to Culicoides‐borne diseases. [ABSTRACT FROM AUTHOR]

Epizootic haemorrhagic disease virus (EHDV) is an arbovirus transmitted by Culicoides biting midges that has recently emerged in Europe. Here, the basic reproduction ratio (R0) was used to quantify the transmission of EHDV and its dependence on temperature for cattle and deer. Using data from the published literature the parameters needed to calculate R0 were estimated with Bayesian methods to incorporate uncertainty in the calculations. The Sobol method of sensitivity analysis was used to determine the parameters having the greatest influence on R0 and, hence, to identify important data gaps. Depending on the strain, the maximum R0 for EHDV varied from 0.7 to 2.5 in cattle and 1.3 to 4.3 in deer. The maximum R0 occurred at temperatures between 22 and 25°C, while the lowest temperature at which R0 exceeded one was between 16 and 20°C. The sensitivity analysis identified the threshold temperature for virus replication, the probability of transmission from host to vector and the vector- to- host ratio as the most important parameters influencing R0. Furthermore, there are only limited data on EHDV in European deer species and on transmission in wildlife and at the livestock/wildlife interface. These data gaps should be the focus of future research.

Culicoides are vectors that can transmit many different pathogens to mammals — including humans, and domestic and wild animals — and birds. In order to take preventive measures against any vector-borne disease, it is important to gather information on both the host and vector species. Culicoides species are mainly mammalophilic, ornithophilic or ornithophilic/mammalophilic, but females have also been found to occasionally feed on engorged insects. A recent systematic review based on three groups of key words investigated Culicoides on farms, and asserted that 92 species (including four not present species) have been reported among cattle in mainland France and Corsica. We have re-evaluated the presence of Culicoides species in cattle in France using the same data of the review. Our data show that only 18 species are reported among cattle. Furthermore, our research used molecular and indirect investigations to analyse Culicoides species that had been feeding on cattle. Our results demonstrate that 45 species feed on cattle out of 92 species present in France. The paper discusses the relevance of data in the literature when investigating hosts of Culicoides species.

IntroductionInsect bite hypersensitivity (IBH) is an IgE-mediated allergic dermatitis of horses caused by bites of Culicoides spp., sharing some common features with human atopic dermatitis. Allergen immunotherapy (AIT) using Culicoides whole-body extracts has limited efficacy. This study aimed to evaluate AIT with a pool of major Culicoides recombinant allergens in a prospective, double-blinded, placebo-controlled study.MethodsThe IBH lesion score was assessed during a pre-treatment year and first treatment year (May–October) in 17 horses and in May and July of a second treatment year. Nine horses were immunized subcutaneously 3× with a combination of nine r-allergens (20 μg each/injection) in alum and monophosphoryl lipid A (MPLA). Eight horses received a placebo. The immunization was repeated twice the following year. The specific antibody response to one of the AIT Culicoides r-allergens was assessed.ResultsIn the first treatment year, the decrease in average IBH lesion score was significantly larger in the AIT compared to the placebo group, with 67% of the AIT group and 25% of the placebo horses reaching >50% improvement of the average IBH lesion score. The response to the AIT was enhanced in the 2nd treatment year when 89% of the AIT vs. 14% of the placebo horses showed an improvement (p ≤ 0.01). IgG antibodies of all subclasses were induced, with IgG4/7 showing the most significant differences between groups. The post-AIT sera showed IgE blocking activity.DiscussionAIT using only a few injections of small amounts of r-allergens in alum and MPLA as immunomodulators seems a promising approach for the treatment of insect bite allergy.

Abstract Introduction Culicoides Latreille biting midges are vectors of high concern as they can transmit serious veterinary diseases such as bluetongue virus or epizootic haemorrhagic disease virus, among others. Little is known about these vectors in Galicia, so a comprehensive literature review and an intensive monitoring were carried out in the region. Material and methods The Autonomous Community of Galicia was sampled through three different vector surveillance projects between 2004 and 2023. A total of 239 sampling points were deployed alongside the Galician territory. In addition, a literature review of Culicoides in Galicia related content was made by consulting several digital repositories. Results A total of 33 species of Culicoides belonging to 8 subgenera were identified. Among them, 15 are considered or suspected to be potential vectors of several pathogens of medical and/or veterinary interest. In addition, 20 of them are reported for the first time in the region. Updated distribution maps of the Culicoides biting midges of Galicia were provided, including several notes regarding their ecology and relevance for both public health and animal welfare. Conclusions The present work is one of the most complete works made at regional level in Spain to date. As Galicia's economy relies heavily on livestock farming, this work will provide a solid baseline in order to develop new research lines in the future regarding prevention to vector‐borne diseases.

Abstract Background As a primary vector of bluetongue virus (BTV) in the US, seasonal abundance and diel flight activity of Culicoides sonorensis has been documented, but few studies have examined how time of host-seeking activity is impacted by environmental factors. This knowledge is essential for interpreting surveillance data and modeling pathogen transmission risk. Methods The diel host-seeking activity of C. sonorensis was studied on a California dairy over 3 years using a time-segregated trap baited with CO2. The relationship between environmental variables and diel host-seeking activity (start, peak, and duration of activity) of C. sonorensis was evaluated using multiple linear regression. Fisher’s exact test and paired-sample z-test were used to evaluate the seasonal difference and parity difference on diel host-seeking activity. Results Host-seeking by C. sonorensis began and reached an activity peak before sunset at a higher frequency during colder months relative to warmer months. The time that host-seeking activity occurred was associated low and high daily temperature as well as wind speed at sunset. Colder temperatures and a greater diurnal temperature range were associated with an earlier peak in host-seeking. Higher wind speeds at sunset were associated with a delayed peak in host-seeking and a shortened duration of host-seeking. Parous midges reached peak host-seeking activity slightly later than nulliparous midges, possibly because of the need for oviposition by gravid females before returning to host-seeking. Conclusions This study demonstrates that during colder months C. sonorensis initiates host-seeking and reaches peak host-seeking activity earlier relative to sunset, often even before sunset, compared to warmer months. Therefore, the commonly used UV light-baited traps are ineffective for midge surveillance before sunset. Based on this study, surveillance methods that do not rely on light trapping would provide a more accurate estimate of host-biting risk across seasons. The association of environmental factors to host-seeking shown in this study can be used to improve modeling or prediction of host-seeking activity. This study identified diurnal temperature range as associated with host-seeking activity, suggesting that Culicoides may respond to a rapidly decreasing temperature by shifting to an earlier host-seeking time, though this association needs further study. Graphical Abstract

(1) Background: Epizootic hemorrhagic disease virus (EHDV) and bluetongue virus (BTV) are orbiviruses that cause hemorrhagic disease (HD) with significant economic and population health impacts on domestic livestock and wildlife. In the United States, white-tailed deer (Odocoileus virginianus) are particularly susceptible to these viruses and are a frequent blood meal host for various species of Culicoides biting midges (Diptera: Ceratopogonidae) that transmit orbiviruses. The species of Culicoides that transmit EHDV and BTV vary between regions, and larval habitats can differ widely between vector species. Understanding how midges are distributed across landscapes can inform HD virus transmission risk on a local scale, allowing for improved animal management plans to avoid suspected high-risk areas or target these areas for insecticide control. (2) Methods: We used occupancy modeling to estimate the abundance of gravid (egg-laden) and parous (most likely to transmit the virus) females of two putative vector species, C. stellifer and C. venustus, and one species, C. haematopotus, that was not considered a putative vector. We developed a universal model to determine habitat preferences, then mapped a predicted weekly midge abundance during the HD transmission seasons in 2015 (July–October) and 2016 (May–October) in Florida. (3) Results: We found differences in habitat preferences and spatial distribution between the parous and gravid states for C. haematopotus and C. stellifer. Gravid midges preferred areas close to water on the border of well and poorly drained soil. They also preferred mixed bottomland hardwood habitats, whereas parous midges appeared less selective of habitat. (4) Conclusions: If C. stellifer is confirmed as an EHDV vector in this region, the distinct spatial and abundance patterns between species and physiological states suggest that the HD risk is non-random across the study area. [ABSTRACT FROM AUTHOR]

Yunnan province in China shares its borders with three neighboring countries: Myanmar, Vietnam, and Laos. The region is characterized by a diverse climate and is known to be a suitable habitat for various arthropods, including midges which are notorious for transmitting diseases which pose significant health burdens affecting both human and animal health. A total of 431,100 midges were collected from 15 different locations in the border region of Yunnan province from 2015 to 2020. These midges were divided into 37 groups according to the collection year and sampling site. These 37 groups of midges were then homogenized to extract nucleic acid. Metatranscriptomics were used to analyze their viromes. Based on the obtained cytochrome C oxidase I gene (COI) sequences, three genera were identified, including one species of Forcipomyia, one species of Dasyhelea, and twenty-five species of Culicoides. We identified a total of 3199 viruses in five orders and 12 families, including 1305 single-stranded positive-stranded RNA viruses (+ssRNA) in two orders and seven families, 175 single-stranded negative-stranded RNA viruses (−ssRNA) in two orders and one family, and 1719 double-stranded RNA viruses in five families. Six arboviruses of economic importance were identified, namely Banna virus (BAV), Japanese encephalitis virus (JEV), Akabane virus (AKV), Bluetongue virus (BTV), Tibetan circovirus (TIBOV), and Epizootic hemorrhagic disease virus (EHDV), all of which are capable, to varying extents, of causing disease in humans and/or animals. The survey sites in this study basically covered the current distribution area of midges in Yunnan province, which helps to predict the geographic expansion of midge species. The complexity and diversity of the viral spectrum carried by midges identified in the study calls for more in-depth research, which can be utilized to monitor arthropod vectors and to predict the emergence and spread of zoonoses and animal epidemics, which is of great significance for the control of vector-borne diseases. [ABSTRACT FROM AUTHOR]

Simple Summary: The number of leishmaniasis cases caused by autochthonous Leishmania (Mundinia) martiniquensis and Leishmania (Mundinia) orientalis has been continuously growing in Southeast Asia over the decades, particularly in Thailand. Recent evidence has suggested that Culicoides biting midges are the most likely natural vectors of autochthonous leishmaniasis in Thailand. Nevertheless, the epidemiology of vector infection and the genetic variation of Leishmania parasites in endemic areas have remained largely unknown. In this study, we report the high infection prevalence, sympatric circulation, and regional genetic diversity of two Leishmania (Mundinia) parasites in Culicoides spp. caught from the affected community and Tham Phra cave in Chiang Rai Province, Northern Thailand. These results provide us with a more complete understanding of the complexity of leishmaniasis transmission, which will be useful for establishing effective management and control measures for this neglected disease, especially in endemic areas of Northern Thailand. In this research, we elucidated the species composition of Culicoides biting midges, infection prevalence, and genetic diversity of Leishmania parasites circulating in the affected community in Chiang Rai Province, being the most endemic area in Northern Thailand. A total of 146 parous and gravid females, belonging to at least twelve Culicoides species in five subgenera and one species group, were trapped from three collection sites with an overall Leishmania prevalence of 26.7% (39/146). Leishmania was detected, using ITS1-PCR, in C. mahasarakamense (15), C. guttifer (11), C. (Trithecoides) spp. (8), C. jacobsoni (2), C. oxystoma (2), and C. orientalis (1). The evidence of Leishmania infection in these last five species represents new records in Northern Thailand. Given a high infection rate in cavernicolous specimens, this indicates an increased risk of parasite exposure when visiting the cave. Using the nanopore amplicon sequencing, L. martiniquensis was ubiquitously identified in all positives, and more than half of these were also co-infected with L. orientalis. The genetic diversity analysis revealed 13 and 17 unique haplotypes for L. martiniquensis and L. orientalis, respectively. Higher haplotype diversity and relatively low nucleotide diversity were observed in both parasite populations, suggesting recent population divergence. Neutrality tests (Tajima's D and Fu and Li's D) showed to be significantly negative, indicating rapid population growth or a selective sweep. Moreover, dominant haplotypes of both Leishmania species were 100% identical to those in all leishmaniasis patients previously reported from Northern Thailand, strongly supporting the imperative role of Culicoides spp. in disease transmission. Essentially, this research provides the first entomological surveillance data representing the sympatric existence, transmission dynamics, and genetic complexity of two autochthonous Leishmania (Mundinia) parasites in several Culicoides species in the endemic area of Northern Thailand. This would contribute to a more complete understanding of the epidemiology of vector infection and facilitate the development of vector control programs to effectively reduce the transmission of this neglected tropical disease in endemic areas of Northern Thailand. [ABSTRACT FROM AUTHOR]

Simple Summary: This study aimed to describe the epidemiological, clinicopathological, and therapeutic aspects of allergic dermatitis caused by Culicoides spp. bites in donkeys. Clinical signs included restlessness and severe itching. Skin lesions were observed across various parts of the body and included areas of alopecia with crusts and serosanguinous exudates. Overall, 378 Culicoides insects were collected and Culicoides ocumarensis Ortiz was identified as the most common species. These findings suggest an association between allergic dermatitis and Culicoides. Additionally, a combination of copaiba oil and a multivitamin emulsion showed therapeutic potential. To date, this is the first study on allergic dermatitis in donkeys in Brazil. An allergy to bites from Culicoides (Diptera: Ceratopogonidae) occurs because of a hypersensitivity reaction caused by the inoculation of insect salivary antigens during the bite, resulting in immune-mediated dermatitis. To the best of our knowledge, no previous studies have focused on allergic dermatitis in donkeys in Brazil. Therefore, this study aimed to describe the epidemiological, clinicopathological, and therapeutic aspects of allergic dermatitis in donkeys and to identify the insects involved in its epidemiology. This study reported the occurrence of dermatitis in 17 animals. The clinical signs were restlessness and severe itching. Skin lesions were found on the head, depigmented areas of the muzzle and cheeks, flanks, pelvic and thoracic limbs, and the scrotal sac. The lesions were characterized by areas of alopecia with crusts accompanied by serosanguineous exudates. Histologically, the lesions were characterized as moderate superficial dermatitis with irregular epidermal acanthosis and pronounced diffuse orthokeratotic hyperkeratosis. In total, 378 Culicoides specimens were collected, with Culicoides ocumarensis Ortiz being the most abundant species. The combined application of copaiba oil and a multivitamin emulsion exhibited potential for topical treatment of allergic dermatitis caused by insect bites in donkeys. Our study revealed an association between allergic dermatitis in donkeys and Culicoides. [ABSTRACT FROM AUTHOR]

Background: Culicoides is a genus of ubiquitous biting midges (Ceratopogonidae). Female midges have blood‐sucking habit. They not only bite and harass humans and animals but also may be an important vector of disease transmission. Therefore, building an animal allergy model caused by Culicoides biting is very beneficial for studying its pathogenesis and exploring the therapeutic methods. Material and method: Kunming mice were used in this study to build the model and sensitised by two‐step injection of midge extracts. Scratching behaviour and histological examination were used to check the immediate and delayed responses. Immunoglobulin E (IgE) and Immunoglobulin G (IgG) were detected using indirect enzyme‐linked immunosorbent assay (ELISA) assay. Splenic cell proliferation and cytokine production were determined using 3‐(4, 5‐dimethylthiazol‐2‐yl)‐2, 5‐diphenyl tetrazolium bromide (MTT) and ELISA assays. The response of cytokine gene expression to midge stimulation was analysed through quantitative real‐time polymerase chain reaction (qPCR). Results: Behavioural results revealed a significant increase in scratching frequency among the midge‐sensitised animals (p < 0.05). Histological examination showed more inflammatory cytokine infiltration at the injection site of midge‐sensitised mice comparing to the ones in the control group. The serum levels of IgE and IgG1 antibodies in the midge‐sensitised group were significantly elevated (p < 0.05). After splenocytes were stimulated in vitro with midge extracts, the midge‐sensitised group's splenocyte count significantly increased in comparison to the control group. The midge‐sensitised group's qPCR data revealed a down‐regulation of tumor necrosis factor alpha (TNF‐α) expression and an increase in the expression of interleukin (IL)‐4, IL‐5, IL‐10 and IL‐13 but not in the control group (p < 0.05). Conclusions: In this study, an animal model of Culicoides–mouse sensitisation was successfully constructed using a two‐step method. The mode of administration of the model was in good agreement with the natural immune pathway, and the immune response induced by the sensitisation of the model was similar to that produced by the bite of a midge. [ABSTRACT FROM AUTHOR]

Background: Culicoides, also known as biting midges, carry pathogens which include Mansonella perstans. Mansonella perstans is a nematode parasite implicated in a number of disease outcomes. Even though a high prevalence of about 75% M. perstans infection has been recorded in some communities in the middle belt of Ghana, and a wide diversity of Culicoides species has been identified, the exact Culicoides species transmitting M. perstans in Ghana has not yet been deciphered. This study therefore aimed at assessing the species diversity of Culicoides and their role in the transmission of M. perstans in the middle belt of Ghana. Methods: Culicoides species were sampled from 11 communities in the Asante-Akim North and Sene West districts in the middle belt of Ghana. Centre for Disease Control (CDC) UV light traps, as well as human bait (i.e. human landing catch and engorged catch) methods were used to assess the species abundance and diversity of Culicoides in the study communities in the wet and dry season. A colorimetric Loop-Mediated Isothermal Amplification (LAMP) assay was performed to assess the vector competence of the various Culicoides species. Results: A total of 4810 Culicoides from 6 species were sampled. These included Culicoides inornatipennis, C. milnei, C. schultzei, C. grahamii, C. neavei, and C. imicola. Culicoides imicola was the most abundant species (56%) followed by C. grahamii (16%). Light traps sampled the most diverse species (6 species). Human landing catch and engorged catch methods identified three anthropophilic species, C. grahamii, C. milnei, and C. inornatipennis, with C. grahamii being the most anthropophilic with a peak biting time between the hours of 5 p.m. to 6 p.m. Generally, there was relatively higher species abundance in the wet than dry season. LAMP assay identified C. grahamii as the potential vector for M. perstans transmission in the middle belt of Ghana. Conclusions: For the first time, we have demonstrated that C. grahamii is the potential competent vector for M. perstans transmission in the middle belt of Ghana. It is more abundant in the rainy season and has a peak biting time between the hours of 5 and 6 p.m. [ABSTRACT FROM AUTHOR]

Forcipomyia Meigen, 1818, is the second-largest genus of the family Ceratopogonidae, having medico veterinary and agricultural importance. A few mitogenomes of Ceratopogonidae have previously been available. Here, we aimed to characterize the whole mitogenome of Forcipomyia pulchrithorax Edwards, 1924, collected from Central Anatolia Region of the Türkiye using NGS to contribute to the genetic diversity within this family. The mitogenome of F. pulchrithorax (OR666457) consisted of a circular DNA molecule spanning 15,930 bp and containing 72.7% AT content. It includes 13 proteincoding genes (PCGs), 22 transfer RNA (tRNA) genes, 2 ribosomal RNA (rRNA) genes, and 1 control region. All PCGs exhibited typical ATN start codons and followed the conventional TAN stop codons, except COX1, COX2, COX3, ND1, and ND5, which ended with incomplete codons. The 22 tRNA genes demonstrated the ability to form a cloverleaf structure, except tRNA-Ser1, which lacked DHU arm, similar to most insect mitogenomes. The ML phylogenetic analyses clearly delineated the species of Culicomorpha into well supported monophyletic clades. Forcipomyia pulchrithorax was clustered with the Ceratopogonidae species constituting an outer taxon among the Culicomorpha. The comprehensive mitogenome of F. pulchrithorax will be valuable for future studies focused on the phylogenetic characterization and diversity of Ceratopogonidae within the Culicomorpha. [ABSTRACT FROM AUTHOR]

Epizootic hemorrhagic disease (EHD) is a non-contagious arthropod-transmitted viral disease and a World Organization for Animal Health (WOAH)-listed disease of domestic and wild ruminants since 2008. EHDV is transmitted among susceptible animals by a few species of midges of genus Culicoides. During the fall of 2021, a large outbreak caused by the epizootic hemorrhagic disease virus (EHDV), identified as serotype 8, was reported in Tunisian dairy and beef farms with Bluetongue virus (BTV)-like clinical signs. The disease was detected later in the south of Italy, in Spain, in Portugal and, more recently, in France, where it caused severe infections in cattle. This was the first evidence of EHDV-8 circulation outside Australia since 1982. In this study, we analyzed the epidemiological situation of the 2021–2022 EHDV outbreaks reported in Tunisia, providing a detailed description of the spatiotemporal evolution of the disease. We attempted to identify the eco-climatic factors associated with infected areas using generalized linear models (GLMs). Our results demonstrated that environmental factors mostly associated with the presence of C. imicola, such as digital elevation model (DEM), slope, normalized difference vegetation index (NDVI), and night-time land surface temperature (NLST)) were by far the most explanatory variables for EHD repartition cases in Tunisia that may have consequences in neighboring countries, both in Africa and Europe through the spread of infected vectors. The risk maps elaborated could be useful for disease control and prevention strategies. [ABSTRACT FROM AUTHOR]

Simple Summary: Haemoproteus parasites are one of the most studied avian blood parasites; however, their natural vectors, Culicoides biting midges, have been identified for only a small portion of them. The main reason for that might be due to the existence of a few research groups working with an integrative approach that allows not only the identification of parasite DNA in the insects, but also confirms the presence of the parasite infective stage (sporozoites) using microscopy. In this study, we aimed to identify the natural vectors of Haemoproteus parasites and to determine their prevalence in Culicoides biting midges in four different localities in Lithuania. Almost 2000 parous Culicoides females belonging to 11 species previously registered in the country were processed. The most abundant one was C. pictipennis. Parasite DNA was found in 7.9% of the analyzed biting midges, and sporozoites were present in almost 30% of these insects. The Botanical Garden was the locality with the highest abundance of biting midges and parasite species, as well as the place with the highest number of biting midges containing sporozoites. One new Haemoproteus vector (C. reconditus) was confirmed, and 12 new interactions between Haemoproteus parasite lineages and Culicoides biting midges of different species were identified. This study helps to understand the relationship between Haemoproteus parasites and Culicoides biting midges in the wild. Haemoproteus parasites are the most diverse among Haemosporida. However, their natural vectors (Culicoides) are still poorly investigated and were identified for only a few parasite species and lineages. The application of an integrative approach (insect dissection, microscopic analysis, and molecular-based methods) is necessary in these studies, which have been carried out by a few research groups, mainly in Europe. The aim of this study was (i) to determine the Culicoides species that are naturally infected by Haemoproteus parasites, and which can support its complete sporogonic development, and (ii) to investigate the prevalence of Culicoides species and Haemoproteus parasite lineages in different study sites. In total, 1953 parous Culicoides females, from 11 species, were collected in four different localities in Lithuania and were dissected and analyzed using an integrative approach. The most abundant was C. pictipennis (30.3%). Parasite DNA was found in 7.9% of all investigated Culicoides, of which ~30% had sporozoites in their salivary glands, confirming their vector competence for these parasites. The Botanical Garden presented the highest number of Culicoides parous females, Culicoides species, and parasite lineages, as well as the highest positivity for sporozoites. Culicoides reconditus was confirmed as a natural vector of Haemoproteus parasites, sporozoites of six Haemoproteus lineages were reported for the first time, and 12 new interactions between Haemoproteus parasite lineages and Culicoides species were identified. Haemoproteus parasites seem to be transmitted by a high number of Culicoides species, with C. kibunensis, C. pictipennis, and C. segnis being the most important vectors. [ABSTRACT FROM AUTHOR]

Bluetongue virus (BTV) is a segmented, double-stranded RNA virus transmitted by Culicoides midges that infects ruminants. As global temperatures increase and geographical ranges of midges expand, there is increased potential for BTV outbreaks from incursions of novel serotypes into endemic regions. However, an understanding of the effect of temperature on reassortment is lacking. The objectives of this study were to compare how temperature affected Culicoides survival, virogenesis, and reassortment in Culicoides sonorensis coinfected with two BTV serotypes. Midges were fed blood meals containing BTV-10, BTV-17, or BTV serotype 10 and 17 and maintained at 20 °C, 25 °C, or 30 °C. Midge survival was assessed, and pools of midges were collected every other day to evaluate virogenesis of BTV via qRT-PCR. Additional pools of coinfected midges were collected for BTV plaque isolation. The genotypes of plaques were determined using next-generation sequencing. Warmer temperatures impacted traits related to vector competence in offsetting ways: BTV replicated faster in midges at warmer temperatures, but midges did not survive as long. Overall, plaques with BTV-17 genotype dominated, but BTV-10 was detected in some plaques, suggesting parental strain fitness may play a role in reassortment outcomes. Temperature adds an important dimension to host–pathogen interactions with implications for transmission and evolution. [ABSTRACT FROM AUTHOR]

The emergence of culicoid-transmitted bluetongue and Schmallenberg viruses in several European countries demonstrated the ability of indigenous biting midge species to transmit pathogens. Entomologic research programs identified members of the Obsoletus Group (Culicoides subgenus Avaritia) as keyplayers in disease epidemiology in Europe. However, morphological identification of potential vectors is challenging due to the recent discovery of new genetic variants (haplotypes) of C. obsoletus sensu stricto (s.s.), forming distinct clades. In this study, 4422 GenBank entries of the mitochondrial cytochrome c oxidase subunit I (COI) gene of subgenus Avaritia members of the genus Culicoides were analyzed to develop a conventional multiplex PCR, capable of detecting all vector species and clades of the Western Palearctic in this subgenus. Numerous GenBank entries incorrectly assigned to a species were identified, analyzed and reassigned. The results suggest that the three C. obsoletus clades represent independent species, whereas C. montanus should rather be regarded as a genetic variant of C. obsoletus s.s. Based on these findings, specific primers were designed and validated with DNA material from field-caught biting midges which achieved very high diagnostic sensitivity (100%) when compared to an established reference PCR (82.6%). [ABSTRACT FROM AUTHOR]

Abstract Background Bluetongue is a non-contagious viral disease that affects both domestic and wild ruminants. It is transmitted primarily by small hematophagous Diptera belonging to the genus Culicoides (Diptera: Ceratopogonidae). The current study represents the first molecular investigation into the potential role of Culicoides imicola, Culicoides paolae, Culicoides newsteadi, Culicoides spp., and Culicoides circumscriptus as bluetongue virus (BTV) vectors in Morocco. Additionally, the study aimed to evaluate the vectorial activity of midges during the survey seasons. Methods Parous females of these species were captured from several regions of Morocco (6 out of 12) from 2018 to 2021 using Onderstepoort Veterinary Institute (OVI) traps. A total of 2003 parous female specimens were grouped into 55 batches. The midge body of each batch was dissected into three regions (head, thorax, and abdomen), and these regions were analyzed separately using reverse transcription quantitative polymerase chain reaction (RT-qPCR). Results BTV RNA was detected in 45 out of the 55 batches tested, indicating a positivity rate of 81.8%. The RT-qPCR-positive pools of the studied Culicoides species exhibited high levels of BTV positivity in each body part (head, thorax, and abdomen), confirming the successful replication of the virus within midge bodies. The BTV circulation was substantial across all three survey seasons (spring, summer, and autumn). High infection rates, calculated using the minimum infection rate (MIR) and maximum likelihood estimation (MLE), were observed during the collection seasons, particularly in autumn and spring, and for all investigated Culicoides species, most notably for C. imicola and C. newsteadi. These increased infection rates underscore the significant risk of Culicoides transmitting the BTV in Morocco. Conclusions The detection of BTV positivity in Culicoides spp. (lacking wing spots that allow their differentiation according to morphological identification keys) suggested that other Culicoides species are competent for BTV transmission in Morocco. The study results indicated, for the first time at the molecular level, that C. imicola and C. newsteadi are the primary potential vectors of BTV in Morocco and that C. paolae and C. circumscriptus are strongly implicated in the propagation of bluetongue at the national level. Graphical Abstract

Background: Culicoides are Diptera of the Family Ceratopogonidae commonly called biting midges or nosee-ums with the capacity as biological vectors of pathogens causing notifiable diseases. Objectives: We surveyed to ascertain the presence and species composition of Culicoides species from various locations and catch points in Maiduguri, Nigeria. Methodology: Culicoides insects were trapped from the selected locations and study points using laboratory-designed sticky traps made of wooden cardboard, painted bright yellow, and coated thinly with petroleum jelly (Vaseline®) which serves as an adhesive. Results: The incidence of Culicoides species between August and November in this study revealed a total catch of 34 using the sticky board trapping with Culicoides bolitinos having 32 (94.1%) and C. engubandei 2 (5.9%). Distribution of catches based on study location/foci and/or catch points/loci showed the highest incidence on the University of Maiduguri livestock farm with 11 (32.4%) and drinking troughs with 15 (44.1%). In conclusion, this study has revealed a location cum catch point dependent incidence of Culicoides in Maiduguri, Nigeria. Conclusion: Despite the low incidence of Culicoides species, we established their presence across the different study locations. We recommend that further studies be conducted to ascertain the seasonal presence as well as their preferred feeding host in the study locations.

Vesicular stomatitis (VS) is a viral disease that affects horses, cattle, and swine that is transmitted by direct contact and hematophagous insects. In 2023, a multi-state outbreak of vesicular stomatitis New Jersey virus (VSNJV) occurred in California, Nevada, and Texas, infecting horses, cattle, and rhinoceros. To identify possible insect vectors, we conducted insect surveillance at various locations in San Diego County, CA, including at a wildlife park. CO2 baited traps set from mid-May to mid-August 2023 collected 2357 Culicoides biting midges and 1215 Simulium black flies, which are insect genera implicated in VSNJV transmission. Insects were pooled by species, location, and date, then tested for viral RNA. Nine RNA-positive pools of Culicoides spp. and sixteen RNA-positive pools of Simulium spp were detected. Infectious virus was detected by cytopathic effect in 96% of the RNA-positive pools. This is the first report of VSNJV in wild-caught C. bergi, C. freeborni, C. occidentalis, S. argus, S. hippovorum, and S. tescorum. The vector competency of these species for VSNJV has yet to be determined but warrants examination. Active vector surveillance and testing during disease outbreaks increases our understanding of the ecology and epidemiology of VS and informs vector control efforts.

Bluetongue (BT) is a Culicoides midge-borne hemorrhagic disease affecting cervids and ruminant livestock species, resulting in significant economic losses from animal production and trade restrictions. Experimental animal infections using the α/β interferon receptor knockout IFNAR mouse model and susceptible target species are critical for understanding viral pathogenesis, virulence, and evaluating vaccines. However, conducting experimental vector-borne transmission studies with the vector itself are logistically difficult and experimentally problematic. Therefore, experimental infections are induced by hypodermic injection with virus typically derived from baby hamster kidney (BHK) cells. Unfortunately, for many U.S. BTV serotypes, it is difficult to replicate the severity of the disease seen in natural, midge-transmitted infections by injecting BHK-derived virus into target host animals. Using the IFNAR BTV murine model, we compared the virulence of traditional BHK cell-derived BTV-17 with C. sonorensis midge (W8) cell-derived BTV-17 to determine whether using cells of the transmission vector would provide an in vitro virulence aspect of vector-transmitted virus. At both low and high doses, mice inoculated with W8-BTV-17 had an earlier onset of viremia, earlier onset and peak of clinical signs, and significantly higher mortality compared to mice inoculated with BHK-BTV-17. Our results suggest using a Culicoides W8 cell-derived inoculum may provide an in vitro vector-enhanced infection to more closely represent disease levels seen in natural midge-transmitted infections while avoiding the logistical and experimental complexity of working with live midges.

Background Bluetongue is a non-contagious viral disease that afects both domestic and wild ruminants. It is transmitted primarily by small hematophagous Diptera belonging to the genus Culicoides (Diptera: Ceratopogonidae). The current study represents the frst molecular investigation into the potential role of Culicoides imicola, Culicoides paolae, Culicoides newsteadi, Culicoides spp., and Culicoides circumscriptus as bluetongue virus (BTV) vectors in Morocco. Additionally, the study aimed to evaluate the vectorial activity of midges during the survey seasons. Methods Parous females of these species were captured from several regions of Morocco (6 out of 12) from 2018 to 2021 using Onderstepoort Veterinary Institute (OVI) traps. A total of 2003 parous female specimens were grouped into 55 batches. The midge body of each batch was dissected into three regions (head, thorax, and abdomen), and these regions were analyzed separately using reverse transcription quantitative polymerase chain reaction (RT-qPCR). Results BTV RNA was detected in 45 out of the 55 batches tested, indicating a positivity rate of 81.8%. The RT-qPCRpositive pools of the studied Culicoides species exhibited high levels of BTV positivity in each body part (head, thorax, and abdomen), confrming the successful replication of the virus within midge bodies. The BTV circulation was substantial across all three survey seasons (spring, summer, and autumn). High infection rates, calculated using the minimum infection rate (MIR) and maximum likelihood estimation (MLE), were observed during the collection seasons, particularly in autumn and spring, and for all investigated Culicoides species, most notably for C. imicola and C. newsteadi. These increased infection rates underscore the signifcant risk of Culicoides transmitting the BTV in Morocco. Conclusions The detection of BTV positivity in Culicoides spp. (lacking wing spots that allow their diferentiation according to morphological identifcation keys) suggested that other Culicoides species are competent for BTV transmission in Morocco. The study results indicated, for the frst time at the molecular level, that C. imicola and C. newsteadi are the primary potential vectors of BTV in Morocco and that C. paolae and C. circumscriptus are strongly implicated in the propagation of bluetongue at the national level. [ABSTRACT FROM AUTHOR]

Introduction: Bluetongue virus (BTV) is an arthropod-borne Orbivirus that is almost solely transmitted by Culicoides biting midges and causes a globally important haemorrhagic disease, bluetongue (BT), in susceptible ruminants. Infection with BTV is characterised by immunosuppression and substantial lymphopenia at peak viraemia in the host. Methods: In this study, the role of cell-mediated immunity and specific T-cell subsets in BTV pathogenesis, clinical outcome, viral dynamics, immune protection, and onwards transmission to a susceptible Culicoides vector is defined in unprecedented detail for the first time, using an in vivo arboviral infection model system that closely mirrors natural infection and transmission of BTV. Individual circulating CD4+, CD8+, or WC1+ gd T-cell subsets in sheep were depleted through the administration of specific monoclonal antibodies. Results: The absence of cytotoxic CD8+ T cells was consistently associated with less severe clinical signs of BT, whilst the absence of CD4+ andWC1+ gd T cells both resulted in an increased clinical severity. The absence of CD4+ T cells also impaired both a timely protective neutralising antibody response and the production of IgG antibodies targeting BTV non-structural protein, NS2, highlighting that the CD4+ Tcell subset is important for a timely protective immune response. T cells did not influence viral replication characteristics, including onset/dynamics of viraemia, shedding, or onwards transmission of BTV to Culicoides. We also highlight differences in T-cell dependency for the generation of immunoglobulin subclasses targeting BTV NS2 and the structural protein, VP7. Discussion: This study identifies a diverse repertoire of T-cell functions during BTV infection in sheep, particularly in inducing specific anti-viral immune responses and disease manifestation, and will support more effective vaccination strategies. [ABSTRACT FROM AUTHOR]

Bluetongue Virus (BTV) and Epizootic Hemorrhagic Disease Virus (EHDV) are Orbiviruses primarily transmitted by their biological vector, Culicoides spp. Latreille, 1809 (Diptera: Ceratopogonidae). These viruses can infect a diverse range of vertebrate hosts, leading to disease outbreaks in domestic and wild ruminants worldwide. This study, conducted at the Belo Horizonte Municipal Parks and Zoobotany Foundation (FPMZB-BH), Minas Gerais, Brazil, focused on Orbivirus and its vectors. Collections of Culicoides spp. were carried out at the FPMZB-BH from 9 December 2021 to 18 November 2022. A higher prevalence of these insects was observed during the summer months, especially in February. Factors such as elevated temperatures, high humidity, fecal accumulation, and proximity to large animals, like camels and elephants, were associated with increased Culicoides capture. Among the identified Culicoides spp. species, Culicoides insignis Lutz, 1913, constituted 75%, and Culicoides pusillus Lutz, 1913, 6% of the collected midges, both described as competent vectors for Orbivirus transmission. Additionally, a previously unreported species in Minas Gerais, Culicoides debilipalpis Lutz, 1913, was identified, also suspected of being a transmitter of these Orbiviruses. The feeding preferences of some Culicoides species were analyzed, revealing that C. insignis feeds on deer, Red deer (Cervus elaphus) and European fallow deer (Dama dama). Different Culicoides spp. were also identified feeding on humans, raising concerns about the potential transmission of arboviruses at the site. In parallel, 72 serum samples from 14 susceptible species, including various Cervids, collected between 2012 and 2022 from the FPMZB-BH serum bank, underwent Agar Gel Immunodiffusion (AGID) testing for BTV and EHDV. The results showed 75% seropositivity for BTV and 19% for EHDV. Post-testing analysis revealed variations in antibody presence against BTV in a tapir and a fallow deer and against EHDV in a gemsbok across different years. These studies confirm the presence of BTV and EHDV vectors, along with potential virus circulation in the zoo. Consequently, implementing control measures is essential to prevent susceptible species from becoming infected and developing clinical diseases. [ABSTRACT FROM AUTHOR]

Bluetongue virus (BTV) is a segmented, double-stranded RNA orbivirus listed by the World Organization for Animal Health and transmitted by Culicoides biting midges. Segmented viruses can reassort, which facilitates rapid and important genotypic changes. Our study evaluated reassortment in Culicoides sonorensis midges coinfected with different ratios of BTV-10 and BTV-17. Midges were fed blood containing BTV-10, BTV-17, or a combination of both serotypes at 90:10, 75:25, 50:50, 25:75, or 10:90 ratios. Midges were collected every other day and tested for infection using pan BTV and cox1 (housekeeping gene) qRT-PCR. A curve was fit to the ∆Ct values (pan BTV Ct—cox1 Ct) for each experimental group. On day 10, the midges were processed for BTV plaque isolation. Genotypes of the plaques were determined by next-generation sequencing. Pairwise comparison of ∆Ct curves demonstrated no differences in viral RNA levels between coinfected treatment groups. Plaque genotyping indicated that most plaques fully aligned with one of the parental strains; however, reassortants were detected, and in the 75:25 pool, most plaques were reassortant. Reassortant prevalence may be maximized upon the occurrence of reassortant genotypes that can outcompete the parental genotypes. BTV reassortment and resulting biological consequences are important elements to understanding orbivirus emergence and evolution. [ABSTRACT FROM AUTHOR]