Culicoides Midges Research Articles

Genomic Detection of Schmallenberg Virus, Israel.

We discuss genomic detection of Schmallenberg virus in both Culicoides midges and affected ruminants during June 2018–December 2019, demonstrating its circulation in Israel. This region is a geographic bridge between 3 continents and may serve as an epidemiologic bridge for potential Schmallenberg virus spread into Asia.

Predicting temperature-dependent transmission suitability of bluetongue virus in livestock

The transmission of vector-borne diseases is governed by complex factors including pathogen characteristics, vector–host interactions, and environmental conditions. Temperature is a major driver for many vector-borne diseases including Bluetongue viral (BTV) disease, a midge-borne febrile disease of ruminants, notably livestock, whose etiology ranges from mild or asymptomatic to rapidly fatal, thus threatening animal agriculture and the economy of affected countries. Using modeling tools, we seek to predict where the transmission can occur based on suitable temperatures for BTV. We fit thermal performance curves to temperature-sensitive midge life-history traits, using a Bayesian approach. We incorporate these curves into S(T), a transmission suitability metric derived from the disease’s basic reproductive number, R_0. This suitability metric encompasses all components that are known to be temperature-dependent. We use trait responses for two species of key midge vectors, Culicoides sonorensis and Culicoides variipennis present in North America. Our results show that outbreaks of BTV are more likely between 15^{circ } C and 34^{circ }hbox { C}, with predicted peak transmission risk at 26 ^circ C. The greatest uncertainty in S(T) is associated with the following: the uncertainty in mortality and fecundity of midges near optimal temperature for transmission; midges’ probability of becoming infectious post-infection at the lower edge of the thermal range; and the biting rate together with vector competence at the higher edge of the thermal range. We compare three model formulations and show that incorporating thermal curves into all three leads to similar BTV risk predictions. To demonstrate the utility of this modeling approach, we created global suitability maps indicating the areas at high and long-term risk of BTV transmission, to assess risk and to anticipate potential locations of disease establishment.

Detection of arboviruses in Culicoides (Diptera: Ceratopogonidae) collected from animal farms in the border areas of Yunnan Province, China

Biting midges of the genus Culicoides (order Diptera, family Ceratopogonidae) are potential biological vectors for the transmission of certain arboviruses among humans, livestock, and wild animals. This study collected a total of 405 Culicoides individuals from seven animal farms located in five counties in the border areas of Yunnan Province, China, and examined the Culicoides species composition and the major arboviruses carried by the Culicoides species. The collected Culicoides were classified into seven species with variable abundances: Culicoides arakawae (5.43%, 22/405), Culicoides homotomus (1.23%, 5/405), Culicoides obsoletus (19.75%, 80/405), Culicoides orientalis (17.28%, 70/405), Culicoides oxystoma (29.38%, 119/405), Culicoides peregrinus (5.68%, 23/405), and Culicoides nipponensis (21.23%, 86/405). Among the seven species, C. oxystoma and C. nipponensis were distributed in all the five counties with abundances of 13.33–44.87% and 10.00–46.83%, respectively, suggesting that these were the dominant species of Culicoides widespread on animal farms in the border areas. PCR was used to detect major arboviruses in the collected Culicoides specimens, including bluetongue virus (BTV), Japanese encephalitis virus, Dengue virus, Zika virus, African swine fever virus, and African horse sickness virus. Among the tested viruses, only BTV serotype 1 was tested positive in C. oxystoma specimens collected from a buffalo farm. Culicoides oxystoma was the dominant species on animal farms in the sampled areas, but it has not previously been documented as positive for BTV in China. The current results thus suggest that C. oxystoma could be an important vector for BTV transmission in these border areas, which, however, needs to be confirmed by further comprehensive experiments. Overall, the present study provides the first profile of Culicoides species on animal farms in the China, Vietnam, and Myanmar border areas, establishes the prevalence of arboviruses carried by these Culicoides species, and suggests the vector potential of C. oxystoma species for the transmission of BTV.

Comparison of recombinant Culicoides allergens produced in different expression systems for IgE serology of insect bite hypersensitivity in horses of different origins

Insect bite hypersensitivity (IBH) is an allergic dermatitis of horses, an IgE-mediated reaction to Culicoides midges. Causative Culicoides spp. are not indigenous in Iceland resulting in high prevalence of IBH in horses born in Iceland and exported as compared to Icelandic horses born in a Culicoides rich environment. The aims were (i) to compare IgE levels in sera of IBH-affected horses born in Iceland (n = 47) with horses of the Icelandic breed (n = 23) and of other breeds (n = 27) born in Culicoides infested area; (ii) to investigate if barley could be a useful production system of allergens for IBH immunoassays.IgE binding in sera was tested by ELISA on two recombinant Culicoides allergens, rCul n 3 and rCul n 4, each produced in E. coli, insect cells and barley. Significantly more IgE was detected against all allergens in sera from IBH-affected compared to healthy horses. Icelandic-born Icelandic horses stand out with higher IgE levels against the allergens and higher area under the curve (AUC) on rCul n 4 as compared to the European-born horses. The barley and E.coli produced allergens had very similar performance in distinguishing between IBH-affected and healthy horses.

Experimental transmission of Leishmania (Mundinia) parasites by biting midges (Diptera: Ceratopogonidae).

Leishmania parasites, causative agents of leishmaniasis, are currently divided into four subgenera: Leishmania, Viannia, Sauroleishmania and Mundinia. The recently established subgenus Mundinia has a wide geographical distribution and contains five species, three of which have the potential to infect and cause disease in humans. While the other Leishmania subgenera are transmitted exclusively by phlebotomine sand flies (Diptera: Psychodidae), natural vectors of Mundinia remain uncertain. This study investigates the potential of sand flies and biting midges of the genus Culicoides (Diptera: Ceratopogonidae) to transmit Leishmania parasites of the subgenus Mundinia. Sand flies (Phlebotomus argentipes, P. duboscqi and Lutzomyia migonei) and Culicoides biting midges (Culicoides sonorensis) were exposed to five Mundinia species through a chicken skin membrane and dissected at specific time intervals post bloodmeal. Potentially infected insects were also allowed to feed on ear pinnae of anaesthetized BALB/c mice and the presence of Leishmania DNA was subsequently confirmed in the mice using polymerase chain reaction analyses. In C. sonorensis, all Mundinia species tested were able to establish infection at a high rate, successfully colonize the stomodeal valve and produce a higher proportion of metacyclic forms than in sand flies. Subsequently, three parasite species, L. martiniquensis, L. orientalis and L. sp. from Ghana, were transmitted to the host mouse ear by C. sonorensis bite. In contrast, transmission experiments entirely failed with P. argentipes, although colonisation of the stomodeal valve was observed for L. orientalis and L. martiniquensis and metacyclic forms of L. orientalis were recorded. This laboratory-based transmission of Mundinia species highlights that Culicoides are potential vectors of members of this ancestral subgenus of Leishmania and we suggest further studies in endemic areas to confirm their role in the lifecycles of neglected pathogens.

Seasonal Abundance of Culicoides at Yongsan US Army Garrison (USAG) and Camp Humphreys USAG, Republic of Korea, 2010-2013 and 2014-2017.

Biting midges (Culicoides: Ceratopogonidae) were collected using New Jersey light traps at Yongsan US Army Garrison (USAG;urban), Seoul Metropolitan city and Camp Humphreys USAG (rural), Pyeongtaek, Gyeonggi-do (province), Republic of Korea, from May-October 2010–2013 and 2015–2017, to determine species composition and seasonal distribution patterns in urban and rural habitats. A total of 9,958 female (53.85%) and 8,533 male (46.15%) Culicoides comprising 16 species were collected. Overall, the most commonly collected species was Culicoides arakawae (74.3%), followed by C. circumscriptus (16.2%), C. kibunensis (2.5%), C. nasuensis (2.2%), C. clavipalpis (1.4%), and C. pallidulus (1.3%), while the remaining 10 species accounted for <2.1% of all Culicoides spp. collected. The 2 predominant species collected were C. circumscriptus (47.4%) and C. arakawae (33.4%) at Yongsan, and C. arakawae (90.4%) and C. circumscriptus (3.9%) at Camp Humphreys. The seasonal abundance of these 2 species varied between years and between sites but on average peaked in August–September for C. arakawae and June–July for C. circumscriptus. Annual variations in abundance were observed for most species collected during this study. Unusually high proportions of male specimens were observed for most species at both sites which may be due to the use of the New Jersey trap.

Exposure of Culicoides sonorensis to Enzootic Strains of Bluetongue Virus Demonstrates Temperature- and Virus-Specific Effects on Virogenesis.

Bluetongue virus (BTV) is a segmented RNA virus transmitted by Culicoides midges. Climatic factors, animal movement, vector species, and viral mutation and reassortment may all play a role in the occurrence of BTV outbreaks among susceptible ruminants. We used two enzootic strains of BTV (BTV-2 and BTV-10) to explore the potential for Culicoides sonorensis, a key North American vector, to be infected with these viruses, and identify the impact of temperature variations on virogenesis during infection. While BTV-10 replicated readily in C. sonorensis following an infectious blood meal, BTV-2 was less likely to result in productive infection at biologically relevant exposure levels. Moreover, when C. sonorensis were co-exposed to both viruses, we did not detect reassortment between the two viruses, despite previous in vitro findings indicating that BTV-2 and BTV-10 are able to reassort successfully. These results highlight that numerous factors, including vector species and exposure dose, may impact the in vivo replication of varying BTV strains, and underscore the complexities of BTV ecology in North America.

Spatio-temporal modelling of Culicoides Latreille (Diptera: Ceratopogonidae) populations on Reunion Island (Indian Ocean)

BackgroundReunion Island regularly faces outbreaks of bluetongue and epizootic hemorrhagic diseases, two insect-borne orbiviral diseases of ruminants. Hematophagous midges of the genus Culicoides (Diptera: Ceratopogonidae) are the vectors of bluetongue (BTV) and epizootic hemorrhagic disease (EHDV) viruses. In a previous study, statistical models based on environmental and meteorological data were developed for the five Culicoides species present in the island to provide a better understanding of their ecology and predict their presence and abundance. The purpose of this study was to couple these statistical models with a Geographic Information System (GIS) to produce dynamic maps of the distribution of Culicoides throughout the island.MethodsBased on meteorological data from ground weather stations and satellite-derived environmental data, the abundance of each of the five Culicoides species was estimated for the 2214 husbandry locations on the island for the period ranging from February 2016 to June 2018. A large-scale Culicoides sampling campaign including 100 farms was carried out in March 2018 to validate the model.ResultsAccording to the model predictions, no husbandry location was free of Culicoides throughout the study period. The five Culicoides species were present on average in 57.0% of the husbandry locations for C. bolitinos Meiswinkel, 40.7% for C. enderleini Cornet & Brunhes, 26.5% for C. grahamii Austen, 87.1% for C. imicola Kieffer and 91.8% for C. kibatiensis Goetghebuer. The models also showed high seasonal variations in their distribution. During the validation process, predictions were acceptable for C. bolitinos, C. enderleini and C. kibatiensis, with normalized root mean square errors (NRMSE) of 15.4%, 13.6% and 16.5%, respectively. The NRMSE was 27.4% for C. grahamii. For C. imicola, the NRMSE was acceptable (11.9%) considering all husbandry locations except in two specific areas, the Cirque de Salazie—an inner mountainous part of the island—and the sea edge, where the model overestimated its abundance.ConclusionsOur model provides, for the first time to our knowledge, an operational tool to better understand and predict the distribution of Culicoides in Reunion Island. As it predicts a wide spatial distribution of the five Culicoides species throughout the year and taking into consideration their vector competence, our results suggest that BTV and EHDV can circulate continuously on the island. As further actions, our model could be coupled with an epidemiological model of BTV and EHDV transmission to improve risk assessment of Culicoides-borne diseases on the island.Graphical

Peripheral blood basophils are the main source for early interleukin-4 secretion upon in vitro stimulation with Culicoides allergen in allergic horses

Interleukin-4 (IL-4) is a key cytokine secreted by type 2 T helper (Th2) cells that orchestrates immune responses during allergic reactions. Human and mouse studies additionally suggest that basophils have a unique role in the regulation of allergic diseases by providing initial IL-4 to drive T cell development towards the Th2 phenotype. Equine Culicoides hypersensitivity (CH) is a seasonal immunoglobulin E (IgE)-mediated allergic dermatitis in horses in response to salivary allergens from Culicoides (Cul) midges. Here, we analyzed IL-4 production in peripheral blood mononuclear cells (PBMC) of CH affected (n = 8) and healthy horses (n = 8) living together in an environment with natural Cul exposure. During Cul exposure when allergic horses had clinical allergy, IL-4 secretion from PBMC after stimulation with Cul extract was similar between healthy and CH affected horses. In contrast, allergic horses had higher IL-4 secretion from PBMC than healthy horses during months without allergen exposure. In addition, allergic horses had increased percentages of IL-4+ cells after Cul stimulation compared to healthy horses, while both groups had similar percentages of IL-4+ cells following IgE crosslinking. The IL-4+ cells were subsequently characterized using different cell surface markers as basophils, while very few allergen-specific CD4+ cells were detected in PBMC after Cul extract stimulation. Similarly, IgE crosslinking by anti-IgE triggered basophils to produce IL-4 in all horses. PMA/ionomycin consistently induced high percentages of IL-4+ Th2 cells in both groups confirming that T cells of all horses studied were capable of IL-4 production. In conclusion, peripheral blood basophils produced high amounts of IL-4 in allergic horses after stimulation with Cul allergens, and allergic horses also maintained higher basophil percentages throughout the year than healthy horses. These new findings suggest that peripheral blood basophils may play a yet underestimated role in innate IL-4 production upon allergen activation in horses with CH. Basophil-derived IL-4 might be a crucial early signal for immune induction, modulating of immune responses towards Th2 immunity and IgE production.

Physicochemical factors affecting the diversity and abundance of Afrotropical Culicoides species in larval habitats in Senegal

Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) are the biological vectors of arboviruses of global importance in animal health. We characterized the physicochemical parameters that determine the density and composition of the main Culicoides species of veterinary interest in larval habitats of the Niayes region of Senegal. For this purpose, we combined larval and substrate sampling in the field in different habitat types with adult emergence and physicochemical analyses in the laboratory. Three major habitat types were identified, conditioning the predominant species of Culicoides and pH and the amount of organic matter were positively correlated with the abundance of larvae and emerging Culicoides, as opposed to salinity. The diversity of emerging Culicoides was positively correlated with pH while it was negatively correlated with salinity. Culicoides distinctipennis was the predominant species in the larval habitat group of freshwater lake edges. In the larval habitat group of pond and puddle edges, C. oxystoma and C. nivosus were predominant; both species were again most abundant in the larval habitat group of saltwater lake edges. These variabilities in physicochemical parameters support the distribution of different Culicoides species in different habitat groups. These results make it possible to implement effective, selective and environmental-friendly control measures but also to improve current models for estimating the abundance of adult vector populations at a local scale.

Culicoides imicola (Biting Midge)

The biting midge Culicoides imicola is a small, haematophagous fly that plays a key role in the biological transmission of viral diseases including bluetongue and African horse sickness (AHS) that inflict damage on both subsistence and intensive livestock as well as companion animals and wildlife. Within the subgenus Avaritia, the monophyletic Imicola group includes ten species, although cryptic diversity may hide the existence of additional species closely related to C. imicola. C. imicola has the broadest geographic range in the genus Culicoides, including most of the African continent, the Mediterranean basin and parts of southern Europe, the Middle East, India, and Southeast Asia to Taiwan.

Vaccination With Recombinant Adenoviruses Expressing the Bluetongue Virus Subunits VP7 and VP2 Provides Protection Against Heterologous Virus Challenge.

Bluetongue virus (BTV) is the causative agent of a disease that affects domestic and wild ruminants and leads to critical economic losses. BTV is an arbovirus from the Reoviridae family that is typically transmitted by the bite of infected Culicoides midges. BTV possesses multiple serotypes (up to 28 have been described), and immunity to one serotype offers little cross-protection to other serotypes. The design of vaccines that provide protection across multiple serotypes is therefore highly desirable to control this disease. We previously reported that a recombinant replication-defective human adenovirus serotype 5 (Ad5) that expresses the VP7 inner core protein of BTV serotype 8 (Ad5VP7-8) induced T-cell responses and provided protection. In the present work, we evaluated as BTV vaccine the combination of Ad5VP7-8 with another recombinant Ad5 that expresses the outer core protein VP2 from BTV-1 (Ad5VP2-1). The combination of Ad5VP2-1 and Ad5VP7-8 protected against homologous BTV challenge (BTV-1 and BTV-8) and partially against heterologous BTV-4 in a murine model. Cross-reactive anti-BTV immunoglobulin G (IgG) were detected in immunized animals, but no significant titers of neutralizing antibodies were elicited. The Ad5VP7-8 immunization induced T-cell responses that recognized all three serotypes tested in this study and primed cytotoxic T lymphocytes specific for VP7. This study further confirms that targeting antigenic determinant shared by several BTV serotypes using cellular immunity could help develop multiserotype BTV vaccines.

High dispersal capacity of Culicoides obsoletus (Diptera: Ceratopogonidae), vector of bluetongue and Schmallenberg viruses, revealed by landscape genetic analyses

BackgroundIn the last two decades, recurrent epizootics of bluetongue virus and Schmallenberg virus have been reported in the western Palearctic region. These viruses affect domestic cattle, sheep, goats and wild ruminants and are transmitted by native hematophagous midges of the genus Culicoides (Diptera: Ceratopogonidae). Culicoides dispersal is known to be stratified, i.e. due to a combination of dispersal processes occurring actively at short distances and passively or semi-actively at long distances, allowing individuals to jump hundreds of kilometers.MethodsHere, we aim to identify the environmental factors that promote or limit gene flow of Culicoides obsoletus, an abundant and widespread vector species in Europe, using an innovative framework integrating spatial, population genetics and statistical approaches. A total of 348 individuals were sampled in 46 sites in France and were genotyped using 13 newly designed microsatellite markers.ResultsWe found low genetic differentiation and a weak population structure for C. obsoletus across the country. Using three complementary inter-individual genetic distances, we did not detect any significant isolation by distance, but did detect significant anisotropic isolation by distance on a north-south axis. We employed a multiple regression on distance matrices approach to investigate the correlation between genetic and environmental distances. Among all the environmental factors that were tested, only cattle density seems to have an impact on C. obsoletus gene flow.ConclusionsThe high dispersal capacity of C. obsoletus over land found in the present study calls for a re-evaluation of the impact of Culicoides on virus dispersal, and highlights the urgent need to better integrate molecular, spatial and statistical information to guide vector-borne disease control.Graphical

Emerging threat of bluetongue in Eastern and North-Eastern Indian states

Albeit bluetongue (BT) in India is endemic to southern and western parts, eastern and north-eastern parts of the country didn't experience any outbreak or reported active disease. However, sero-epidemiology indicates bluetongue virus (BTV) is in circulation in animals and vectors of eastern and north-eastern states of India. Moreover, in recent years BTV has been isolated from sheep, Culicoides midges and goat from this region that substantiate the notion. All the epidemiological components of BT, viz. susceptible hosts, suitable climatic conditions for propagation of vectors, competent vectors are present that pose emerging disease threat in eastern and north-eastern parts of India. Hence, further surveillance and preparedness are recommended to avoid future BT outbreak, if any.

Diversity, Abundance and Host Preference of Culicoides (Diptera: Ceratopogonidae), Potential Vectors and Nuisance Insect in Zoo Park Košice, Slovakia

Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) were captured in the course of three seasons (2017–2020) in the Zoo Košice. The total number of captured individuals was 5,667 biting midges, and 12 species of Culicoides were identified, while the most frequently captured and the most populous species were C. obsoletus/C. scoticus, C. furcillatus, and C. festivipennis. The most frequent hosts on which the biting midges were blood-feeding were humans (Homo sapiens), Burchell's zebra (Equus quagga burchellii), and domestic cow (Bos taurus). This is the first study to confirm a wide range of biting midges species in a zoo which are capable of using not only animals as their hosts, but also humans, in whom a bite may induce an allergic reaction. More than 80% of the fauna of biting midges in the Zoo Košice represented Culicoides obsoletus/C. scoticus, which are in the Central Europe regarded as the potential vectors of arboviruses, to which primarily ruminants are highly susceptible.

Diversity, Abundance and Host Preference of Culicoides (Diptera: Ceratopogonidae), Potential Vectors and Nuisance Insect in Zoo Park Košice, Slovakia

Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) were captured in the course of three seasons (2017–2020) in the Zoo Košice. The total number of captured individuals was 5,667 biting midges, and 12 species of Culicoides were identified, while the most frequently captured and the most populous species were C. obsoletus/C. scoticus, C. furcillatus, and C. festivipennis. The most frequent hosts on which the biting midges were blood-feeding were humans (Homo sapiens), Burchell's zebra (Equus quagga burchellii), and domestic cow (Bos taurus). This is the first study to confirm a wide range of biting midges species in a zoo which are capable of using not only animals as their hosts, but also humans, in whom a bite may induce an allergic reaction. More than 80% of the fauna of biting midges in the Zoo Košice represented Culicoides obsoletus/C. scoticus, which are in the Central Europe regarded as the potential vectors of arboviruses, to which primarily ruminants are highly susceptible.

Blood meal analysis: host-feeding patterns of biting midges (Diptera, Ceratopogonidae, Culicoides Latreille) in Slovakia.

Biting midges of the genus Culicoides are vectors of important pathogens affecting domestic and wild animals and have played a major role in the re-emergence of new outbreaks of bluetongue (BTV) and Schmallenberg (SBV) viruses in Europe. To determine vector-host specificity, trophic preference from blood meal analysis is of major importance in the surveillance of arthropod-borne diseases. Of 28,752 specimens collected, we identified 17 Culicoides species and investigated a total of 48 host sequences from the blood meals. Culicoides obsoletus/C. scoticus, C. dewulfi, C. pulicaris, C. lupicaris, C. punctatus, C. newsteadi, C. riethi, and C. furcillatus were found to feed on mammals (cattle, horses, and humans), birds (domestic chickens), small rodents (Apodemus flavicollis), and hares (Lepus europaeus). To our knowledge, this is the first study investigating trophic preferences of Culicoides spp. in Slovakia. This study demonstrated that Culicoides species are able to feed on domesticated host vertebrates as well as birds, rodents, and humans.

Prevalence and Age Grading of Two Common Culicoides species, in Farms of Warangal Districts of Telangana State, South India

The prevalence and age grading of Culicoides midges from two districts of Warangal in Telangana State, South India was studied considering the species as pests and vectors. The study revealed the species composition, the abundance, host preference and age grading of Culicoides. The morphological characters were used for specific identification of biting midges. The midges were collected from two different farms from Warangal rural and urban districts (cattle and sheep) using CDC-UV light trap connected with suction fan for the period of one year (September 2017 to August 2018). A total of 28,000 flies were collected in which 25,000 (83.33%) were female and 3,000 (16.66%) were males. The species included 10,500 (58.33%) C. oxystoma, 7,500 (41.66 %) C. imicola from cattle farms 10,000 flies from sheep farms including C. imicola 6,500 (65%) and C. oxystoma 3,500 (35%). The age grading of Culicoides revealed significant proportion of parous females which play an important role in transmission of Blue Tongue disease virus.

Viral Vector Vaccines against Bluetongue Virus.

Bluetongue virus (BTV), the prototype member of the genus Orbivirus (family Reoviridae), is the causative agent of an important livestock disease, bluetongue (BT), which is transmitted via biting midges of the genus Culicoides. To date, up to 29 serotypes of BTV have been described, which are classified as classical (BTV 1–24) or atypical (serotypes 25–27), and its distribution has been expanding since 1998, with important outbreaks in the Mediterranean Basin and devastating incursions in Northern and Western Europe. Classical vaccine approaches, such as live-attenuated and inactivated vaccines, have been used as prophylactic measures to control BT through the years. However, these vaccine approaches fail to address important matters like vaccine safety profile, effectiveness, induction of a cross-protective immune response among serotypes, and implementation of a DIVA (differentiation of infected from vaccinated animals) strategy. In this context, a wide range of recombinant vaccine prototypes against BTV, ranging from subunit vaccines to recombinant viral vector vaccines, have been investigated. This article offers a comprehensive outline of the live viral vectors used against BTV.

Investigation of bovine ephemeral fever virus transmission by putative dipteran vectors under experimental conditions

BackgroundBovine ephemeral fever virus (Rhabdoviridae: Ephemerovirus) (BEFV) causes bovine ephemeral fever (BEF), an economically important disease of cattle and water buffalo. Outbreaks of BEF in Africa, Australia, Asia and the Middle East are characterized by high rates of morbidity and highly efficient transmission between cattle hosts. Despite this, the vectors of BEFV remain poorly defined.MethodsColony lines of biting midges (Culicoides sonorensis) and mosquitoes (Aedes aegypti, Culex pipiens and Culex quinquefasciatus) were infected with a strain of BEFV originating from Israel by feeding on blood–virus suspensions and by intrathoracic inoculation. In addition, in vivo transmission of BEFV was also assessed by allowing C. sonorensis inoculated by the intrathoracic route to feed on male 6 month-old Holstein-Friesian calves.ResultsThere was no evidence of BEFV replication within mosquitoes fed on blood/virus suspensions for mosquitoes of any species tested for each of the three colony lines. In 170 C. sonorensis fed on the blood/virus suspension, BEFV RNA was detected in the bodies of 13 individuals and in the heads of two individuals, indicative of fully disseminated infections and an oral susceptibility rate of 1.2%. BEFV RNA replication was further demonstrated in all C. sonorensis that were inoculated by the intrathoracic route with virus after 5, 6 or 7 days post-infection. Despite this, transmission of BEFV could not be demonstrated when infected C. sonorensis were allowed to feed on calves.ConclusionsNo evidence for infection or dissemination of BEFV (bovine/Israel/2005-6) in mosquitoes of three different species was found. Evidence was found for infection of C. sonorensis by the oral route. However, attempts to transmit BEFV to calves from infected C. sonorensis failed. These results highlight the challenge of defining the natural vector of BEFV and of establishing an in vivo transmission model. The results are discussed with reference to the translation of laboratory-based studies to inference of vector competence in the field.Graphical abstract