Comparing Culicoides biting rates in horses and cattle in The Netherlands: potential relevance to African horse sickness epidemiology

To determine relative host preference rates and to establish whether a pair of animals in close proximity (between–host interaction) modified Culicoides attack and abundance profiles, compared to those tethered in isolation (host independence), Culicoides midges were pooted hourly from two sets of experimental animals: (1) a heifer cow and Fjord horse tethered close together (5 m apart), and (2) a heifer and Fjord tethered in isolation (45 m apart). Over 12 days, 570 3‐min observations yielded 23 090 midges, representing 24 species. Approximately 95% belonged to the Culicoides obsoletus (Meigen) complex (two species), Culicoides dewulfi Goetghebuer, Culicoides chiopterus (Meigen), Culicoides punctatus (Meigen), Culicoides pulicaris (L.), and Culicoides achrayi Kettle & Lawson (Diptera: Ceratopogonidae) combined. There was no evidence for between‐host interaction. Mean Culicoides species‐specific attack rates did not differ between animal species, except that C. chiopterus was 7× more abundant on the legs of the heifer compared to the horse, and C. dewulfi twice as abundant on the upper half of the horse compared to the heifer. By contrast, mean species–specific biting rates of the C. obsoletus complex, C. chiopterus, C. dewulfi, C. punctatus, and C. pulicaris midges were 5×, 100×, 1.7×, 2×, and 2.5× lower in the horses compared to the heifers, respectively. It is not clear why high Culicoides attack rates observed in the horses do not convert into high biting rates as seen in cattle; this should be a subject for future research. In light of its apparent predilection for equids, the ability of C. dewulfi to replicate African horse sickness virus (AHSV) extrinsically should be investigated in the laboratory.     

Revealing potential bridge vectors for BTV and SBV: a study on Culicoides blood feeding preferences in natural ecosystems in Spain

Several species of Culicoides (Diptera: Ceratopogonidae) are vectors of pathogens, such as the bluetongue (BTV) and Schmallenberg (SBV) viruses, which cause important diseases in domestic and wild ruminants. As wild ruminants can contribute to overwintering and epizootics of both diseases, knowledge of the host‐feeding behaviour of Culicoides in natural ecosystems is important to better understand their epidemiology. Blood‐engorged Culicoides females trapped in natural areas inhabited by different wild ruminant species were genetically analysed to identify host species. The origin of bloodmeals was identified in 114 females of 14 species of Culicoides. A total of 104 (91.1%) Culicoides fed on mammals and 10 (8.9%) on birds. The most abundant host identified was red deer (66.7%), followed by humans (13%) and fallow deer (6.1%). Eleven of the 14 species of Culicoides fed exclusively on mammalian hosts. Among them, five are mammalophilic species considered to be important BTV and/or SBV vectors. The results of the present study confirm that Culicoides imicola, Culicoides obsoletus, Culicoides scoticus, Culicoides pulicaris and Culicoides punctatus fed on wild ruminants, and therefore support the hypothesis that these species can act as bridge vectors by facilitating the circulation of pathogens between wild and domestic ruminant communities.     

Culicoides biting midges and mosquito fauna at three dog and cat shelters in rural and periurban areas in Northern Spain

The diversity and abundance of Culicoides (Diptera: Ceratopogonidae) and mosquitoes (Diptera: Culicidae) were studied in three animal protection centres (APCs) in Northern Spain between 1 July and 31 October 2018. Four miniature suction CDC light traps (two UV and two standard incandescent bulb traps, both types baited and non‐baited with CO₂) were placed in each APC to compare their efficiency in the collection of these Diptera groups. A total of 1176 biting midges (14 species), 224 mosquitoes (8 species) and 1 black fly were collected and identified by both morphological and molecular approaches. The Culicoides obsoletus complex (C. obsoletus/C. scoticus) accounted for 58.2% of the total collection within the Ceratopogonidae family, whereas Culex pipiens/Cx. torrentium comprised 76.8% of the Culicidae. The input of CO₂ in light traps proved largely ineffective in improving the collections of both Diptera groups. UV‐light traps were 7.8 and 2.2 times more effective than incandescent light traps in trapping Culicoides and mosquitoes, respectively. Seasonal dynamics differed between both Diptera taxa but captures of both taxa were significantly larger at the beginning of the summer. The epidemiological relevance of the most prevalent species is also discussed.     

Emergence of Culicoides obsoletus group species from farm‐associated habitats in Germany

Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) may transmit several arboviruses to ruminant livestock. The species of the Obsoletus group are considered to be among the most important vectors of bluetongue virus (BTV) in northern Europe. As agricultural environments offer suitable habitats for the development of their immature stages, the emergence of adult Culicoides from potential breeding sites was investigated at 20 cattle farms throughout Germany in 2012 and 2013. In analyses of species‐specific habitat preferences and relationships between Culicoides abundance in breeding substrates and their physicochemical characteristics, dungheaps emerged as the most important substrate for the development of Culicoides obsoletus sensu stricto (s.s.) (Meigen), whereas Culicoides chiopterus (Meigen) and Culicoides dewulfi Goetghebuer were generally restricted to cowpats. A decreasing pH value was associated with a higher abundance or a higher probability of observing these three species. Furthermore, the abundance of C. obsoletus s.s. was positively related to increasing moisture. Dungheaps were very productive breeding sites for this species and are therefore suggested as a target for potential control measures.     

Limited attractant range of the black‐light suction trap for the capture of Culicoides biting midges (Diptera: Ceratopogonidae)

The suction light trap (LT) is a standard tool used to capture Culicoides biting midges, when estimating abundances, and mapping species ranges. The exact range of attraction of the LT is in dispute, however, with several studies indicating the range to vary widely, between 4 and 50 m. In this study, the attractant range of a LT, set at various distances (0, 1, 2, 3, 5 and 8 m) from a cow tethered at pasture, is assessed in two trials conducted on a dairy farm in the Netherlands in 2013 and 2014. In the first trial, sweep‐netting and light‐trapping was performed concurrently. Although a Spearman Rank correlation of 0.89 (P = 0.03) showed the two sets of abundance results to be highly correlated, species prevalence differed significantly according to the method used: the Culicoides obsoletus complex dominated the LT observations, whereas Culicoides chiopterus and Culicoides dewulfi were more abundant in those obtained by sweep‐net, due to the former taxon being largely nocturnal, the latter diurnal. There is a distinct negative logarithmic relationship between Culicoides abundance and distance from the vertebrate host, with midge abundance, in the LT, increasing significantly only when it is placed very close to the bait animal, at a distance of 0–1 m. The sharp decay function in the number of Culicoides caught with increasing distance from the LT, limits the comparison of LT surveillance data obtained under various settings, even from within a climatically homogeneous geographical region such as north‐western Europe.     

Culicoides (Diptera: Ceratopogonidae) and livestock in the Netherlands: comparing host preference and attack rates on a Shetland pony,a dairy cow,and a sheep

Culicoides (Diptera: Ceratopogonidae) host preferences and attack rates were quantified in early summer at a dairy farm in the Netherlands using livestock tethered at pasture. Midges were aspirated hourly over seven consecutive hours (17:00–23:00) from a dairy cow, a Shetland pony, and a sheep and correspondingly yielded seventeen, thirteen, and nine species. Of the 14,181 midges obtained, approximately 95% belonged to the C. obsoletus complex, C. dewulfi, C. chiopterus, and C. punctatus that together include all proven or potential vectors for arboviral diseases in livestock in northwestern Europe. On average, 7.6 and 3.5 times more Culicoides were collected, respectively, from the cow and the Shetland pony than from the sheep. In descending order of abundance, the C. obsoletus complex, C. dewulfi, and C. chiopterus dominated attacks on all three hosts, whereas C. punctatus and C. pulicaris favored only the two larger hosts. Irrespective of the host species involved, the three body regions attracted the same component species, C. chiopterus favoring the legs, C. punctatus and C. achrayi the belly, and the C. obsoletus complex, C. dewulfi, and C. pulicaris the head, back, and flanks. That known and potential vectors for animal diseases feed indiscriminately on a broad range of mammal hosts means that all major livestock species, including equines, are rendered susceptible to one or more Culicoides‐borne pathogens.     

Genetic structure and population dynamics of the biting midges Culicoides obsoletus and Culicoides scoticus: implications for the transmission and maintenance of bluetongue

Culicoides species belonging to the Obsoletus complex (Diptera: Ceratopogonidae) have been indicated as primary bluetongue (BT) vectors in many European countries and their possible involvement in the maintenance and overwintering of BT viruses has been suggested, even in regions where Culicoides imicola Keiffer is the main vector. The Obsoletus complex includes two predominant taxa, Culicoides obsoletus (Meigen) and Culicoides scoticus Downes & Kettle. However, the role played by each species in the epidemiology of BT is still unknown. Taxonomic identification is mainly based on the morphology of male genitalia and the lack of other reliable diagnostic features makes the screening of trap‐collected vector populations, mainly females, particularly difficult. Although molecular markers have facilitated species identification, little information is yet available on the biology, abundance and population dynamics of the two taxa. The aim of this work was to investigate the genetic profile and temporal distribution of C. obsoletus and C. scoticus by using isozyme electrophoresis applied to adult midges, collected weekly at two selected farms in southern Sardinia. A total of nine enzyme loci were analysed and five of them provided diagnostic allozyme markers (Hk, Mdh, Pgi, Idh‐1 and Idh‐2). Nei's genetic distance between the two taxa was in the range of other well‐separated taxa (D = 1.792), supporting their status as true species. Culicoides scoticus represented almost 61% of the 562 specimens analysed; its genetic structure was characterized by a very low level of intra‐population variation (mean heterozygosity H_e = 0.019) and higher genetic divergence between populations (F_ST = 0.0016) than in C. obsoletus. The latter species had significantly more heterozygotes (H_e = 0.123), a higher percentage of polymorphic loci, and no inter‐population differentiation (F_ST?0). We suggest that different biological and ecological constraints, such as breeding habitat requirements, may contribute to shaping the genetic profiles of C. scoticus and C. obsoletus. However, enough gene flow was maintained between populations of each species as no spatial and temporal structuring was sustained by Fisher's exact probability test (P > 0.5). The seasonal distributions of C. scoticus and C. obsoletus only partially overlapped: both species were mainly found early in the year, when the main vector, C. imicola, was present in low numbers, and peaked in abundance in April and May. Culicoides scoticus was predominant until May, decreased rapidly in the following months and increased again in winter, whereas C. obsoletus decreased more slowly and was still present in early summer. Consequently, C. scoticus may be a good candidate for playing a role in the transmission and maintenance of BT virus in Sardinia, as well as in other Mediterranean countries, during the months of late winter and early spring when the seroconversion of sentinel animals is still occurring in the absence of the main vector.     

The Mondrian matrix: Culicoides biting midge abundance and seasonal incidence during the 2006–2008 epidemic of bluetongue in the Netherlands

During the northern Europe epidemic of bluetongue (BT), Onderstepoort‐type blacklight traps were used to capture Culicoides Latreille (Diptera: Ceratopogonidae) biting midges weekly between November 2006 and December 2008 on 21 livestock farms in the Netherlands. Proven and potential vectors for the bluetongue virus (BTV) comprised almost 80% of the midges collected: the Obsoletus complex, constituting C. obsoletus (Meigen) and C. scoticus Downes & Kettle (44.2%), C. dewulfi Goetghebuer (16.4%), C. chiopterus (Meigen) (16.3%) and C. pulicaris (Linnaeus) (0.1%). Half of the 24 commonest species of Culicoides captured completed only one (univoltine) or two (bivoltine) generations annually, whereas multivoltine species (including all BTV vectors) cycled through five to six generations (exceeding the one to four generations calculated in earlier decades). Whether this increment signals a change in the phenology of northern Europe Culicoides or simply is an adaptive response that manifests during warmer episodes, thus heightening periodically the incursive potential of midge‐borne arboviruses, remains to be clarified. Culicoides duddingstoni Kettle & Lawson, C. grisescens Edwards, C. maritimus Kieffer, C. pallidicornis Kieffer and C. riethi Kieffer are new records for the biting midge fauna of the Netherlands. It is suggested that C. punctatus (Meigen) be added to the European list of vector Culicoides.     

Diel activity and preferred landing sites in Culicoides biting midges attacking Fjord horses

In the summer of 2014, in the central part of The Netherlands, Culicoides spp. (Diptera: Ceratopogonidae) attack rates, biting rates, and preferred landing sites were determined for a pair of Fjord horses maintained permanently at pasture in an area devoid of cattle. Eleven body regions of the horses were screened for midges, each region sampled randomly for 5 min using a handheld mouth aspirator (pooter). Observations were confined to the hour immediately before and after sunset. Culicoides spp. were obtained from every body region, of which the four most abundant species – Culicoides chiopterus (Meigen), Culicoides punctatus (Meigen), the species complex Culicoides obsoletus (Meigen), and Culicoides dewulfi Goetghebuer – all were proven or potential vectors for arboviral diseases in livestock. Culicoides spp. activity was distinctly bimodal across the day, surging at sunset and 1 h after sunrise. Midges were inactive between 11:00 and 16:00 hours, these hours marking the time of day when horses can be pastured most safely but, thereafter, to avoid escalating attacks, would have to be stabled protectively. Around sunset, the mean attack rate of the four most abundant species ranged from 3.0 to 11.7 midges per min; of these, C. dewulfi and C. chiopterus were reared out of the dung of experimental horses. The Netherlands is home to the world's densest horse population (11 per km²), of which half are estimated to stay outdoors permanently with no access to protective housing. In the absence of a preventive vaccination policy, it is difficult to envisage how horses in northern Europe will be protected from infection during an outbreak of a Culicoides‐transmitted disease like African horse sickness.     

Phylogenetic relationships of the subgenus AvaritiaFox, 1955 including Culicoides obsoletus (Diptera, Ceratopogonidae) in Italy based on internal transcribed spacer 2 ribosomal DNA sequences

Abstract. Biting midges of the genus Culicoides include vector species for orbiviral diseases, such as bluetongue and African horse sickness. Although the Afro-Asiatic species C. imicola is the major vector of bluetongue in the Mediterranean basin, recent outbreaks in regions where C. imicola is absent has incriminated other Culicoides, including those belonging to the Obsoletus Complex of the subgenus Avaritia Fox, 1955. The classical taxonomy of this species complex is unclear and this stimulated the molecular analysis of twenty Culicoides populations sampled from eighteen localities across Italy. Ribosomal internal transcribed spacer 2 sequences were used to characterize the intra- and interspecific variation between Italian members of the Obsoletus Complex and related species, by means of an analysis of molecular variance and phylogenetic analyses. Although morphological differentiation is often extremely difficult, the molecular analysis clearly demonstrated a high degree of divergence between most species. The study showed that at least seven species of the subgenus Avaritia occur in Italy; these are C. obsoletus, C. scoticus, C. montanus, C. dewulfi, C. imicola and two species that could not be identified with certainty, but one of which is similar to C. chiopterus. Finally, a simple polymerase chain reaction assay was developed that rapidly discriminates between four members of the Obsoletus Complex in Italy, a prerequisite for vector identification.     

Barcoding of biting midges in the genus Culicoides: a tool for species determination

Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) are insect vectors of economically important veterinary diseases such as African horse sickness virus and bluetongue virus. However, the identification of Culicoides based on morphological features is difficult. The sequencing of mitochondrial cytochrome oxidase subunit I (COI), referred to as DNA barcoding, has been proposed as a tool for rapid identification to species. Hence, a study was undertaken to establish DNA barcodes for all morphologically determined Culicoides species in Swedish collections. In total, 237 specimens of Culicoides representing 37 morphologically distinct species were used. The barcoding generated 37 supported clusters, 31 of which were in agreement with the morphological determination. However, two pairs of closely related species could not be separated using the DNA barcode approach. Moreover, Culicoides obsoletus Meigen and Culicoides newsteadi Austen showed relatively deep intraspecific divergence (more than 10 times the average), which led to the creation of two cryptic species within each of C. obsoletus and C. newsteadi. The use of COI barcodes as a tool for the species identification of biting midges can differentiate 95% of species studied. Identification of some closely related species should employ a less conserved region, such as a ribosomal internal transcribed spacer.     

The dying of the light: crepuscular activity in Culicoides and impact on light trap efficacy at temperate latitudes

The light trap is the tool of choice for conducting large‐scale Culicoides (Diptera: Ceratopogonidae) vector surveillance programmes. Its efficacy is in doubt, however. To assess this, hourly changes in Culicoides activity over the 24‐h diel were determined comparatively by way of light trapping and aerial sweeping, and correlated against light intensity. In the Netherlands, sweeping around cattle at pasture revealed that, in early summer, Culicoides are active throughout the diel, and that their abundance peaks during the crepuscular period and falls to a low during the brightest hours of the day. By contrast, the light trap was able to accumulate Culicoides only at night (i.e. after illuminance levels had dropped to 0 lux and midge activity had begun to decline). Although Culicoides chiopterus and species of the Culicoides obsoletus complex were similarly abundant around livestock, they differed critically in their hours of peak activity, being largely diurnal and nocturnal, respectively. This polarity helps to explain why, routinely, the C. obsoletus complex dominates light trap collections and C. chiopterus does not. Inability to accumulate Culicoides at light intensity levels above 0 lux means that, at ever‐higher latitudes, particularly beyond 45° N, the progressive northward lengthening of the twilight period will have an increasingly adverse impact upon the efficacy of the light trap as a vector surveillance tool.     

Indoor and outdoor winter activity of Culicoides biting midges,vectors of bluetongue virus,in Italy

Indoor and outdoor winter activity of Culicoides spp. (Diptera: Ceratopogonidae) in central Italy was investigated in order to evaluate whether indoor activity might account for the overwintering of bluetongue virus, as has been hypothesized by some authors. Weekly Culicoides collections were performed at three farms over three consecutive winter seasons. At each farm, two black‐light traps were operated simultaneously, indoors and outdoors. Culicoides were identified using both morphological and molecular means. The Culicoides obsoletus group accounted for 98.2% of sampled specimens. Within this group, C. obsoletus s.s. accounted for 56.8% and Culicoides scoticus for 43.2% of samples. Nulliparous, parous and engorged females were caught throughout the entire winter, both indoors and outdoors. At times, indoor catch sizes outnumbered outdoor collections. A significant inverse correlation was found between minimum temperature and the proportion of indoor Culicoides of the total midge catch, thus indicating that lower outdoor temperatures drive Culicoides midges indoors. High rates of engorged females were recorded indoors, possibly as the result of the propensity of C. obsoletus females to feed indoors. Higher proportions of parous females were found in indoor than in outdoor catches, indicating higher survival rates indoors and, consequently, higher vectorial capacities of midges sheltering indoors compared with those remaining outdoors.     

Influence of light and kairomone baiting systems on trap collections of biting midges in southern Sweden

Effective surveillance is essential for protecting livestock from Culicoides biting midges and the viruses they transmit. The objective of this study was to determine how the baiting system used in traps (UV, incandescent light, incandescent light with CO₂, and incandescent light with CO₂ and 1‐octen‐3‐ol) influences estimates of midge population abundance, parity, and diel activity. This was achieved through a standardized trapping protocol conducted in three habitats in Sweden. UV light traps caught the most Culicoides species and more C. obsoletus complex females than incandescent light traps. Traps baited with CO₂ plus 1‐octen‐3‐ol caught more female C. impunctatus than incandescent light traps. No consistent effect of bait type was found on C. obsoletus parity rate, as estimated from the proportion of midges with presence or absence of pigmentation. Midge activity, as reflected by trap catches, peaked between ‐3 h and +3 h relative to sunset, with UV traps catching significantly more female C. obsoletus complex and C. impunctatus at and after sunset than before sunset. We conclude that baiting system can influence biting midge collections, even using identical traps. Effective surveillance may require more than one bait type and kairomones to attract species that do not feed exclusively on cattle.     

Larval development and emergence sites of farm‐associated Culicoides in the United Kingdom

Culicoides biting midges (Diptera: Ceratopogonidae) are the biological vectors of internationally important arboviruses of livestock including bluetongue virus (BTV). Information on the habitats used by Culicoides for larval development is valuable for establishing targeted vector control strategies and for improving local scale models of vector abundance. This study combines emergence trap collections of adult Culicoides identified using molecular markers and physiochemical measurements of habitats to investigate larval development sites of Palaearctic Culicoides in South East England. The known range of larval habitats for several Culicoides species is extended and the potential BTV vector species C. obsoletus and C. scoticus are confirmed to co‐occur in many larval habitats. The presence of emerging C. obsoletus was favoured by increasing substrate moisture level [odds ratio (OR) 6.94 (2.30; 20.90)] and substrate pH [OR 4.80 (1.66; 13.90)] [bias‐corrected D_xy: 0.68; area under the curve (AUC): 0.86] rather than any particular larval habitat type, as expected for a species with relatively wide larval habitat preference. Of the newly emerged sub‐genus Avaritia individuals collected, 23% were observed to have a degree of abdominal pigmentation commonly inferred to indicate parity. If consistent across species and locations, this observation represents a potential source of error for age structure analyses of Culicoides populations.     

Trap placement and attractant choice affect capture and create sex and parity biases in collections of the biting midge,Culicoides sonorensis

Culicoides sonorensis Wirth & Jones (Diptera: Ceratopogonidae) is the primary North American vector of bluetongue virus (BTV), which can cause high morbidity and mortality in ruminant livestock or wildlife. Worldwide, most Culicoides surveillance relies on light (usually UV) traps typically placed near animals or larval development sites. However, the trapping method can cause sex, species and parity biases in collections. We collected C. sonorensis from three dairies in California using suction traps baited with CO₂, UV light or CO₂ + UV placed near animals, wastewater ponds, or in fields. Higher numbers of parous females were collected using CO₂ + UV traps, although this difference was only significant on one dairy. UV traps were poor at collecting nulliparous females, but the addition of UV to a trap increased the abundance of males in a collection. Traps set in open fields collected significantly higher numbers of males and females than in either of the other two locations. In some cases, there was a significant interaction between the trap type and site. We discuss the limitations of traditional trapping methodologies for C. sonorensis and make suggestions for vector surveillance.     

Are Bogs Reservoirs for Emerging Disease Vectors? Evaluation of Culicoides Populations in the Hautes Fagnes Nature Reserve (Belgium)

Several species of Culicoides (Diptera: Ceratopogonidae) biting midges serve as biological vectors for the bluetongue virus (BTV) and the recently described Schmallenberg virus (SBV) in northern Europe. Since their recent emergence in this part of the continent, these diseases have caused considerable economic losses to the sheep and cattle industries. Much data is now available that describe the distribution, population dynamics, and feeding habits of these insects. However, little is known regarding the presence of Culicoides in unusual habitats such as peaty marshes, nor their potential vector capacity. This study evaluated Culicoides biting midges present in the bogs of a Belgian nature reserve compared to those residing at a nearby cattle farm. Culicoides were trapped in 2011 at four different sites (broadleaved and coniferous forested areas, open environments, and at a scientific station) located in the Hautes Fagnes Nature Reserve (Belgium). An additional light trap was operated on a nearby cattle farm. Very high numbers of biting midges were captured in the marshy area and most of them (70 to 95%) were Culicoides impunctatus, a potential vector of BTV and other pathogens. In addition, fewer numbers of C. obsoletus/C. scoticus species, C. chiopterus, and C. dewulfi were observed in the bogs compared to the farm. The wet environment and oligotrophic nature of the soil were probably responsible for these changes in the respective populations. A total of 297,808 Culicoides midges belonging to 27 species were identified during this study and 3 of these species (C. sphagnumensis, C. clintoni and C. comosioculatus) were described in Belgium for the first time.     

First detection of endosymbiotic bacteria in biting midges Culicoides pulicaris and Culicoides punctatus,important Palaearctic vectors of bluetongue virus

Heritable bacteria have been highlighted as important components of vector biology, acting as required symbionts with an anabolic role, altering competence for disease transmission, and affecting patterns of gene flow by altering cross compatibility. In this paper, we tested eight U.K. species of Culicoides (Diptera: Ceratopogonidae) midge for the presence of five genera of endosymbiotic bacteria: Cardinium (Bacteroidales: Bacteroidaceae); Wolbachia (Rickettsiales: Rickettsiaceae); Spiroplasma (Entomoplasmatales: Spiroplasmataceae); Arsenophonus (Enterobacteriales: Enterobacteriaceae), and Rickettsia (Rickettsiales: Rickettsiaceae). Cardinium spp. were detected in both sexes of Culicoides pulicaris and Culicoides punctatus, two known vectors of bluetongue virus. Cardinium spp. were not detected in any other species, including the Culicoides obsoletus group, the main vector of bluetongue and Schmallenberg viruses in northern Europe. The other endosymbionts were not detected in any Culicoides species. The Cardinium strain detected in U.K. Culicoides species is very closely related to the Candidatus Cardinium hertigii group C, previously identified in Culicoides species in Asia. Further, we infer that the symbiont is not a sex ratio distorter and shows geographic variation in prevalence within a species. Despite its detection in several species of Culicoides that vector arboviruses worldwide, the absence of Cardinium in the C. obsoletus group suggests that infections of these symbionts may not be necessary to the arboviral vector competence of biting midges.