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We reconstruct the eruptive chronology of the Yucamane–Calientes compound volcano in southern Peru based on extensive fieldwork and a large dataset of geochronological (K-Ar, 40Ar/39Ar, U-Pb, and 14C) and geochemical (major and trace element) analyses. This compound volcano is composed of two edifices that have experienced discontinuous volcanic activity from the middle Pleistocene to the Holocene. The Calientes volcano has been constructed in four successive stages: Calientes I is composed of andesitic lava flows dated at ~500 ka. Subsequently, the Callazas ignimbrite (Calientes II stage) was emplaced ~160–190 ka, followed by the main cone-building stage (Calientes III) at ~130–100 ka. Finally, the Holocene Calientes domes were emplaced and represent the last eruptive products of this edifice. The Yucamane volcano has been constructed in three stages: Yucamane I consists of a succession of andesitic lava flows exposed at the base of the volcano that are older than 40 ka. Yucamane II (~36–30 ka) comprises a thick sequence of block-and-ash-flow deposits that represents an episode of dome growth predating the younger Yucamane cone (Yucamane III) built after 20–25 ka. During the Holocene, Yucamane vulcanian to sub-Plinian activity has emplaced tephra-fall and pyroclastic-density-current deposits. The most recent explosive eruptions occurred ca. 3000 BP and emplaced a tephra-fall and pumice-flow deposits. Most samples from Calientes volcano are andesites and dacites (60.1–67.7 wt% SiO2), while rocks from Yucamane volcano are basaltic-andesites to dacites (53.4–66.9 wt% SiO2). The rocks have a mineral assemblage of plagioclase, amphibole, biotite, orthopyroxene, clinopyroxene, olivine, and Fe-Ti oxides. The analyzed samples are categorized within a high-K, calc-alkaline series. Calientes volcano erupted mostly andesitic magmas, but its history is punctuated by rare eruptions of silica-rich magmas. In contrast, Yucamane volcano follows a different trend characterized by a gradual decrease in silica content through post-glacial time, from the large (VEI 3) sub-Plinian andesitic eruption of ~3 ka to moderate (VEI ≤ 2) vulcanian eruptions of basaltic-andesitic. On the basis of such recurrent and recent (Holocene), low-to-moderate explosive activity, Yucamane must be considered an active and potentially threatening volcano, which may affect the province of Candarave with ~8000 inhabitants.

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International audience; 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.

This study establishes the first faunistic inventory of livestock associated Culicoides (Diptera: Ceratopogonidae) species of Reunion Island (Indian Ocean), where bluetongue and epizootic hemorrhagic disease are regularly recorded. Single night-catches were performed at 41 sites using light suction traps at altitudes ranging from 0 to 1525 m, from March to April 2005. Five species were recorded: Culicoides imicola , Culicoides bolitinos , Culicoides enderleini , Culicoides grahamii , and Culicoides kibatiensis , among which at least the first three species are known to be involved in virus transmission to ruminants and equids. This is the first record of C. bolitinos , C. kibatiensis , and C. enderleini on the island. C. imicola was the most abundant species along the sea coast. C. bolitinos was more abundant inland and on two sites on the east coast. C. kibatiensis and C. grahamii were less abundant than the other three species and limited to two foci. Spatial distribution analysis of the different species showed that C. bolitinos , C. enderleini and C. imicola were collected at low altitudes, while the other two species were found at higher altitude. A morphological identification key for adult females and males is given, as well as cytochrome oxydase subunit I sequences. Phylogenetic reconstructions showed a clear divergence between C. bolitinos from Reunion Island and mainland Africa. This monograph will help to identify the Culicoides species in the poorly known entomological fauna of the south-western Indian Ocean region.

Since bluetongue outbreaks occurred in november 2007 in northern Spain, an intensive trapping program was carried out to study the diversity and abundance of the Culicoides Latreille (Diptera: Ceratopogonidae) species. Eleven sheep farms and seven natural habitats distributed throughout the Basque country region were sampled using UV light CDC traps between 2008-2011. A total of 348,685 Culicoides specimens belonging to 52 species were collected during 1,480 trappings in 24h periods. An updated checklist of these 52 species (including four new records for the Iberian peninsula) is provided from the Basque country. The most abundant species in sheep farms were the two sibling species of the obsoletus complex (81.8% of the total catches): C. obsoletus (Meigen) and C. scoticus Downes and Kettle. Culicoides lupicaris Downes and Kattle was the next most abundant taxon collected in farms (9.7% of the total). Few specimens of Culicoides imicola Kieffer, the bluetongue vector in Mediterranean Basin and some specimens of Culicoides nubeculosus (Meigen), an important candidate of BTV transmission were collected in northeast and northwest farms, respectively. In natural ecosystems, three species, Culicoides festivipennis Kieffer, Culicoides alazanicus Dzhafarov and Culicoides brunnicans Edwards, comprised 48.5% of the total captures. Culicoides obsoletus/C. scoticus was present throughout the year and even during the winter days in temperate Atlantic areas (Gipuzkoa, Biscay) whereas no catches occurred in winter at the southern farms of Alava, where the climate is much colder in that season. The majority of the species were active from March to November with maximum peaks of catches in summer. Most Culicoides obsoletus/C. scoticus specimens were collected outdoors (65.9%). The study shows greater occurrence and abundance of obsoletus and pulicaris complex in sheep farms and in lesser extent in natural ecosystems. Data about diversity, distribution and seasonal dynamics are also reported improving the knowledge of these Culicoides species in terms of surveillance and prevention for future bluetongue outbreaks.

International audience; Feeding success depends on host availability, host defensive reactions and host preferences. Host choice is a critical determinant of the intensity at which pathogens are transmitted. The aim of the current study was to describe host preferences of Palaearctic Culicoides species (Diptera: Ceratopogonidae) Latreille using traps baited with the five different host species of poultry, horse, cattle, sheep and goat. Collections were carried out nightly in July and August 2009 in western France with three replicates of a 5 x 5 randomized Latin square (five sites, five hosts). Moreover, an ultraviolet (UV) light/suction trap was operated during host-baited collections to correlate Culicoides biting rates and UV light/suction trap catches. A total of 660 Culicoides belonging to 12 species, but comprised mainly of Culicoides scoticus Downes and Kettle, Culicoides dewulfi Goetghebuer and Culicoides obsoletus Meigen, were collected on animal baits. Abundance was highest for the horse, which accounted for 95% of all Culicoides caught, representing 10 species. The horse, the largest bait, was the most attractive host, even when abundance data were corrected by weight, body surface or Kleiber's scaling factor. Culicoides obsoletus was the only dominant species attracted by birds. Both C. scoticus and C. dewulfi were collected mainly from the upper body of the horse. Finally, the quantification of host preferences allows for discussion of implications for the transmission of Culicoides-borne pathogens such as bluetongue virus.

Culicoides species (Diptera: Ceratopogonidae) belonging to the Obsoletus and Pulicaris groups are considered to be the main vectors of bluetongue virus (BTV) in non Mediterranean Europe. Selected terrestrial microhabitats (n = 17) on a farm in northern Spain were sampled repeatedly over a year-long period and characterized for use by Culicoides species for immature development. Concurrent use of CDC light traps showed the presence of 37 species and 66,575 specimens of adult Culicoides. A total of 28 species and 11,396 individuals emerged from laboratory-maintained soil samples. Culicoides obsoletus and Culicoides scoticus (pooled as Obsoletus complex) were particularly abundant (comprising 58.6% and 74.5% of the total collections in light traps and emergence traps respectively). Potential key vectors of animal viruses (such as BTV) were found in two main terrestrial types of microhabitats. In the case of C. obsoletus, different types of manure (old and composted manure, manure mixed with organic matter, and fresh manure) produced most of the specimens. In contrast, larvae of C. scoticus and Culicoides lupicaris were associated with soil substantially comprised of rotting leaf litter that included the parasitic plant Lathraea clandestina. Several species, Culicoides festivipennis, Culicoides punctatus and Culicoides brunnicans, were very common in mud at pond margins. Indeed, pond microhabitats and runoff below barn rooflines supported the greatest species richness. In the pond habitat, 49.4% of Culicoides specimens emerged from mud at the water edge, as opposed to 50 cm above (32.4%) and 1 meter above waterline (18%). Similar species richness, but statistically significant differences in abundance, were observed among the four pond microhabitats. Overall, the majority of the specimens were found in the upper layer (0–3 cm), except in manure, where they preferred deeper layers (>6 cm). Maximum peaks of abundance occurred in both light traps and soil samples in summer months, whereas increased captures in autumn were noticed only in light traps. Both trapping systems failed to collect adult Culicoides midges in the coldest months of December, January and February. The literature on immature habitats of species suspected in BTV transmission in Europe, the Pulicaris group and particularly the Obsoletus group, is briefly reviewed.

Background: Certain biting midges species of the genus Culicoides (Diptera: Ceratopogonidae) are vectors of virus to livestock worldwide. Culicoides larval ecology has remained overlooked because of difficulties to identify breeding sites, methodological constraints to collect samples and lack of morphological tools to identify field-collected individuals to the species level. After the 2007 unforeseen outbreaks of African horse sickness virus (AHSV) in Senegal (West Africa), there is a need to identify suitable and productive larval habitats in horse farms for the main Culicoides species to evaluate the implementation of vector control measures or preventive actions.Methods: We investigate twelve putative larval habitats (habitat types) of Culicoides inside and outside of three horse farms in the Niayes area of Senegal using a combination of flotation and emergence methods during four collection sessions.Results: Among the three studied horse farms, three habitat types were found positive for Culicoides larvae: pond edge, lake edge and puddle edge. A total of 1420 Culicoides individuals (519♂/901♀) belonging to ten species emerged from the substrate samples. Culicoides oxystoma (40 %), C. similis (25 %) and C. nivosus (24 %) were the most abundant species and emerged from the three habitat types while C. kingi (5 %) was only retrieved from lake edges and one male emerged from puddle edge. Culicoides imicola (1.7 %) was found in low numbers and retrieved only from pond and puddle edges.Conclusions: Larval habitats identified were not species-specific. All positive larval habitats were found outside the horse farms. This study provides original baseline information on larval habitats of Culicoides species in Senegal in an area endemic for AHSV, in particular for species of interest in animal health. These data will serve as a point of reference for future investigations on larval ecology and larval control measures.

Background Introduction of vector species into new areas represents a main driver for the emergence and worldwide spread of vector-borne diseases. This poses a substantial threat to livestock economies and public health. Culicoides imicola Kieffer, a major vector species of economically important animal viruses, is described with an apparent range expansion in Europe where it has been recorded in south-eastern continental France, its known northern distribution edge. This questioned on further C. imicola population extension and establishment into new territories. Studying the spatio-temporal genetic variation of expanding populations can provide valuable information for the design of reliable models of future spread. Methods Entomological surveys and population genetic approaches were used to assess the spatio-temporal population dynamics of C. imicola in France. Entomological surveys (2–3 consecutive years) were used to evaluate population abundances and local spread in continental France (28 sites in the Var department) and in Corsica (4 sites). We also genotyped at nine microsatellite loci insects from 3 locations in the Var department over 3 years (2008, 2010 and 2012) and from 6 locations in Corsica over 4 years (2002, 2008, 2010 and 2012). Results Entomological surveys confirmed the establishment of C. imicola populations in Var department, but indicated low abundances and no apparent expansion there within the studied period. Higher population abundances were recorded in Corsica. Our genetic data suggested the absence of spatio-temporal genetic changes within each region but a significant increase of the genetic differentiation between Corsican and Var populations through time. The lack of intra-region population structure may result from strong gene flow among populations. We discussed the observed temporal variation between Corsica and Var as being the result of genetic drift following introduction, and/or the genetic characteristics of populations at their range edge. Conclusions Our results suggest that local range expansion of C. imicola in continental France may be slowed by the low population abundances and unsuitable climatic and environmental conditions. Electronic supplementary material The online version of this article (doi:10.1186/s13071-016-1426-4) contains supplementary material, which is available to authorized users.

International audience; Understanding the demographic history and genetic make-up of colonizing species is critical for inferring population sources and colonization routes. This is of main interest for designing accurate control measures in areas newly colonized by vector species of economically important pathogens. The biting midge Culicoides imicola is a major vector of orbiviruses to livestock. Historically, the distribution of this species was limited to the Afrotropical region. Entomological surveys first revealed the presence of C. imicola in the south of the Mediterranean basin by the 1970s. Following recurrent reports of massive bluetongue outbreaks since the 1990s, the presence of the species was confirmed in northern areas. In this study, we addressed the chronology and processes of C. imicola colonization in the Mediterranean basin. We characterized the genetic structure of its populations across Mediterranean and African regions using both mitochondrial and nuclear markers, and combined phylogeographical analyses with population genetics and approximate Bayesian computation. We found a west/east genetic differentiation between populations, occurring both within Africa and within the Mediterranean basin. We demonstrated that three of these groups had experienced demographic expansions in the Pleistocene, probably because of climate changes during this period. Finally, we showed that C. imicola could have colonized the Mediterranean basin in the Late Pleistocene or Early Holocene through a single event of introduction; however, we cannot exclude the hypothesis involving two routes of colonization. Thus, the recent bluetongue outbreaks are not linked to C. imicola colonization event, but rather to biological changes in the vector or the virus.

The identification of blood meal source of arthropod vector species contributes to the understanding of host-vector-pathogen interactions. The aim of the current work was to identify blood meal source in Culicoides biting midge species, biological vectors of internationally important arboviruses of livestock and equids, using a new ecological approach. We examined the correlation between blood meal source identified in engorged Culicoides females collected in a suction light trap and the available vertebrate hosts along four rings (200, 500, 1000 and 2000 m) centered at the trap site and described the foraging range of the three main vector species of veterinary interest present in the study area, Culicoides imicola, Culicoides kingi and Culicoides oxystoma. The study was performed in four sites localized in the Niayes region of Senegal (West Africa) where recent outbreaks of African horse sickness occurred. Blood meal source identification was carried out by species-specific multiplex PCRs with genomic DNA extracted from the abdomen of engorged females collected during nine night collections for twenty-six collections. The four most abundant hosts present in the studied area (horse, cattle, goat and sheep) were surveyed in each ring zone. The blood meal source varied according to Culicoides species and host availability in each site. C. oxystoma and C. imicola females mainly fed on horses readily available at 200 m maximum from the trap location whereas females of C. kingi fed mainly on cattle, at variable distances from the traps (200 to 2000 m). C. oxystoma may also feed on other vertebrates. We discuss the results in relation with the transmission of Culicoides-borne arboviruses and the species dispersion capacities.

International audience; African horse sickness- and bluetongue virus are orbiviruses transmitted by Culicoides biting midges (Diptera: Ceratopogonidae) to horses and to ruminants, respectively. Since the last epizootic outbreak of African horse sickness in 2007 in Senegal, extensive investigations have been undertaken to improve our knowledge on Culicoides species involved locally in the transmission of the virus. The purpose of this study was to compare and quantify the host preferences of potential vectors of these orbiviruses on horse and sheep and to study their circadian rhythm. We found that Culicoides oxystoma and species of the subgenus Avaritia (Culicoides imicola, Culicoides bolitinos and Culicoides pseudopallidipennis) had a preference for horse when compared to sheep (the predicted ratio between horse and sheep was 80 for C oxystoma and 26 for C. imicola), and were mostly crepuscular: C. oxystoma had continuous activity throughout the diel with peaks in numbers collected after sunrise and sunset, while C imicola was mostly nocturnal with peak after sunset. Unexpectedly, species of the subgenus Lasiohelea was also collected during this study. This diurnal biting species was a nuisance pest for both animal species used as bait.

Culicoides biting midges (Diptera: Ceratopogonidae) are important vectors of arboviruses in Africa. Culicoides oxystoma has been recently recorded in the Niayes region of Senegal (West Africa) and its high abundance on horses suggests a potential implication in the transmission of the African horse sickness virus in this region. This species is also suspected to transmit bluetongue virus to imported breeds of sheep. Little information is available on the biology and ecology of Culicoides in Africa. Therefore, understanding the circadian host-seeking activity of this putative vector is of primary importance to assess the risk of the transmission of Culicoides-borne pathogens. To achieve this objective, midges were collected using a sheep-baited trap over two consecutive 24-h periods during four seasons in 2012. A total of 441 Culicoides, belonging to nine species including 418 (94.8 %) specimens of C. oxystoma, were collected. C. oxystoma presented a bimodal circadian host-seeking activity at sunrise and sunset in July and was active 3 h after sunrise in April. Daily activity appeared mainly related to time periods. Morning activity increased with the increasing temperature up to about 27 °C and then decreased with the decreasing humidity, suggesting thermal limits for C. oxystoma activity. Evening activity increased with the increasing humidity and the decreasing temperature, comprised between 20 and 27 °C according to seasons. Interestingly, males were more abundant in our sampling sessions, with similar activity periods than females, suggesting potential animal host implication in the facilitation of reproduction. Finally, the low number of C. oxystoma collected render practical vector-control recommendations difficult to provide and highlight the lack of knowledge on the bio-ecology of this species of veterinary interest. Electronic supplementary material The online version of this article (doi:10.1007/s00436-015-4534-8) contains supplementary material, which is available to authorized users.

Background Culicoides biting midges are biological vectors of internationally important arboviruses of livestock and equines. Insecticides are often employed against Culicoides as a part of vector control measures, but systematic assessments of their efficacy have rarely been attempted. The objective of the present study is to determine baseline susceptibility of multiple Culicoides vector species and populations in Europe and Africa to the most commonly used insecticide active ingredients. Six active ingredients are tested: three that are based on synthetic pyrethroids (alpha-cypermethrin, deltamethrin and permethrin) and three on organophosphates (phoxim, diazinon and chlorpyrifos-methyl). Methods Susceptibility tests were conducted on 29,064 field-collected individuals of Culicoides obsoletus Meigen, Culicoides imicola Kieffer and a laboratory-reared Culicoides nubeculosus Meigen strain using a modified World Health Organization assay. Populations of Culicoides were tested from seven locations in four different countries (France, Spain, Senegal and South Africa) and at least four concentrations of laboratory grade active ingredients were assessed for each population. Results The study revealed that insecticide susceptibility varied at both a species and population level, but that broad conclusions could be drawn regarding the efficacy of active ingredients. Synthetic pyrethroid insecticides were found to inflict greater mortality than organophosphate active ingredients and the colony strain of C. nubeculosus was significantly more susceptible than field populations. Among the synthetic pyrethroids, deltamethrin was found to be the most toxic active ingredient for all species and populations. Conclusions The data presented represent the first parallel and systematic assessment of Culicoides insecticide susceptibility across several countries. As such, they are an important baseline reference to monitor the susceptibility status of Culicoides to current insecticides and also to assess the toxicity of new active ingredients with practical implications for vector control strategies. Electronic supplementary material The online version of this article (doi:10.1186/s13071-015-1042-8) contains supplementary material, which is available to authorized users.

SUMMARYBluetongue is a disease of major economic concern in Europe. Its causative agent, bluetongue virus (BTV), is transmitted by several Culicoides species (mainly Culicoides imicola and Culicoides obsoletus in Europe). The application of insecticides on animals may reduce transmission of BTV, however, no formulation is currently licensed specifically against Culicoides midges. The present study assesses the susceptibility of C. obsoletus to deltamethrin using an adapted World Health Organization (WHO) susceptibility test. Midges were exposed to different dosages of deltamethrin for 1 h, and mortality after 1 h and 24 h was recorded. Results indicated that deltamethrin is highly toxic to C. obsoletus since a dose of 1·33×10−4% was enough to kill 50% of the population (LD50) in 24 h. The deltamethrin concentration needed to kill 90% of the population (LD90) was 5·55×10−4%. The results obtained in the present work could help to create a system that can be used to assess insecticide resistance and susceptibility of Culicoides biting midges.

International audience; Feeding success of free-living hematophagous insects depends on their ability to be active when hosts are available and to reach places where hosts are accessible. When the hematophagous insect is a vector of pathogens, determining the components of host-seeking behavior is of primary interest for the assessment of transmission risk. Our aim was to describe endo/exophagy and circadian host-seeking activity of Palaearctic Culicoides species, which are major biting pests and arbovirus vectors, using drop traps and suction traps baited with four sheep, as bluetongue virus hosts. Collections were carried out in the field, a largely-open stable and an enclosed stable during six collection periods of 24 hours in April/May, in late June and in September/October 2010 in western France. A total of 986 Culicoides belonging to 13 species, mainly C. brunnicans and C. obsoletus, was collected on animal baits. Culicoides brunnicans was clearly exophagic, whereas C. obsoletus was able to enter stables. Culicoides brunnicans exhibited a bimodal pattern of host-seeking activity with peaks just after sunrise and sunset. Culicoides obsoletus was active before sunset in spring and autumn and after sunset in summer, thus illustrating influence of other parameters than light, especially temperature. Description of host-seeking behaviors allowed us to discuss control strategies for transmission of Culicoides-borne pathogens, such as bluetongue virus. However, practical vector-control recommendations are difficult to provide because of the variation in the degree of endophagy and time of host-seeking activity.

Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) are small hematophagous dipterans responsible for transmitting major viruses to livestock such as African horse sickness virus in equids, and pizootic haemorrhagic disease and bluetongue viruses (BTV) in ruminants. Between 1998 and 2005, BTV outbreaks occurred into many countries around the Mediterranean basin associated with the northward extension of C. imicola, which colonized new Mediterranean territories in the past decades due to the global increase of temperatures. In August 2006, BTV serotype 8 (BTV-8) was unexpectedly introduced in the Netherlands and was intensively transmitted by autochthonous Palaearctic Culicoides species, leading in 2007 and 2008 to a major sanitary crisis in whole continental Europe with huge economic losses due to animal movement restrictions. This chapter presents a synthesis of the data gathered through the different entomological surveillance networks implemented in France since 2000 when C. imicola was recorded for the first time in France. These networks aimed at first to monitor the spread and establishment of the invasive C. imicola and then from 2006 onwards, to study the diversity and dynamics of autochthonous Culicoides species. The data collected enable to describe for the first time the spread of C. imicola into French mainland, propose an updated list of Culicoides species for France, describe the Culicoides species distribution and seasonal dynamics, and report assays to identify BTV in field-collected Culicoides during outbreaks. (Texte intégral)

Bluetongue virus is transmitted by Culicoides biting midges (Diptera: Ceratopogonidae). Culicoides associated with livestock were captured using CDC blacklight traps at three BTV-infected farms in Basque Country between November 2007 and December 2008. Twenty-seven and nineteen Culicoides species were collected in outdoor and indoor habitats respectively. Indoor insect community represented 86.1% of the whole captured individual biting midges. Culicoides obsoletus / Culicoides scoticus (two sibling species of the Obsoletus complex) were dominant throughout all months and sexes with maximum phenological peaks in November 2007 and June–July 2008. Culicoides lupicaris was the second most dominant species followed by Culicoides pulicaris (both species of the Pulicaris complex). Few specimens of Culicoides imicola , the principal Afro-Mediterranean vector of BTV, as well as four new species recorded for the Iberian Peninsula, were also collected. BTV was detected by RT-PCR from pools of C. obsoletus / C. scoticus , C. lupicaris and C. pulicaris parous females. dl -Lactic acid significantly attracted more C. obsoletus / C. scoticus females and males, C. lupicaris females, C. pulicaris females and Culicoides punctatus females and males; whereas acetone increased only the captures of Culicoides achrayi .

Bluetongue virus (BTV) is an economically important arbovirus of ruminants transmitted by Culicoides biting midges. Vector control using residual spraying or application to livestock is recommended by many authorities to reduce BTV transmission; however, the impact of these measures in terms of both inflicting mortality on Culicoides and subsequently upon BTV transmission is unclear. This study consisted of a standardized World Health Organization laboratory assay to determine the susceptibility of European Culicoides species to deltamethrin and a field trial based upon allowing individuals of a laboratory strain of Culicoides nubeculosus Meigen to feed upon sheep treated with Butox 7.5 pour-on (a deltamethrin-based topical formulation). Susceptibility in the laboratory trial was higher in colony C. nubeculosus (24-h LC [90]=0.00106%), than in field populations of Culicoides obsoletus Meigen (24-h LC[90]=0.00203%) or Culicoides imicola Kieffer (24-h LC[90]=0.00773%). In the field, the pour-on formulation was tested with a total of 816 C. nubeculosus specimens fed upon on the thigh of treated sheep. The study revealed a maximum mortality rate of 49% at 4 d post application, and duration of lethal effect was predicted to be as short as 10 d, despite testing being carried out with a highly susceptible strain. The reasons for this low efficacy are discussed with reference both to the potential for lack of spread of the active ingredient on the host and feeding patterns of the major potential vector species on the sheep host. Practical implications for vector control strategies during BTV incursions are also detailed.

Background The emergence and massive spread of bluetongue in Western Europe during 2006-2008 had disastrous consequences for sheep and cattle production and confirmed the ability of Palaearctic Culicoides (Diptera: Ceratopogonidae) to transmit the virus. Some aspects of Culicoides ecology, especially host-seeking and feeding behaviors, remain insufficiently described due to the difficulty of collecting them directly on a bait animal, the most reliable method to evaluate biting rates. Our aim was to compare typical animal-baited traps (drop trap and direct aspiration) to both a new sticky cover trap and a UV-light/suction trap (the most commonly used method to collect Culicoides). Methods/results Collections were made from 1.45 hours before sunset to 1.45 hours after sunset in June/July 2009 at an experimental sheep farm (INRA, Nouzilly, Western France), with 3 replicates of a 4 sites × 4 traps randomized Latin square using one sheep per site. Collected Culicoides individuals were sorted morphologically to species, sex and physiological stages for females. Sibling species were identified using a molecular assay. A total of 534 Culicoides belonging to 17 species was collected. Abundance was maximal in the drop trap (232 females and 4 males from 10 species) whereas the diversity was the highest in the UV-light/suction trap (136 females and 5 males from 15 species). Significant between-trap differences abundance and parity rates were observed. Conclusions Only the direct aspiration collected exclusively host-seeking females, despite a concern that human manipulation may influence estimation of the biting rate. The sticky cover trap assessed accurately the biting rate of abundant species even if it might act as an interception trap. The drop trap collected the highest abundance of Culicoides and may have caught individuals not attracted by sheep but by its structure. Finally, abundances obtained using the UV-light/suction trap did not estimate accurately Culicoides biting rate.

After bluetongue (BT) appeared in northern Europe in August 2006 entomological studies were implemented in all five affected Member States (MSs) to establish which species of Culicoides had acted as vectors. The findings can be summarised as follows: (i) C. imicola the principal southern European/African vector of BTV has not penetrated into northern Europe, (ii) three pools of C. obsoletus/C. scoticus and one of C. dewulfi assayed RT-PCR-positive to BTV-8, (iii) in support of these results it was found that both potential vectors had also high parity rates (approximately 40%) indicating increased longevity favouring BTV virogenesis and transmission, (iv) furthermore, C. obsoletus/C. scoticus and C. dewulfi occurred also widely and abundantly on sheep and cattle holdings across the entire affected region, (v) and during the latter part of the season showed strong endophily readily entering livestock buildings in significant numbers to bite the animals inside (endophagy), (vi) which demonstrates that housing at best offers only limited protection to livestock from Culicoides attacks, (vii) in contrast the potential vector C. pulicaris sensu stricto was restricted geographically, was captured rarely, had a low parity rate (10%) and was exophilic indicating it played no role in the outbreak of BT, (viii) the incrimination of C. dewulfi as a novel vector is significant because it breeds in cattle and horse dung this close association raising its vectorial potential, but (ix) problems with its taxonomy (and that of the Obsoletus and Pulicaris species complexes) illustrates the need for morphological and molecular techniques to become more fully integrated to ensure progress in the accurate identification of vector Culicoides, (x) midge densities (as adjudged by light traps) were generally low indicating northern European Culicoides to have a high vector potential and/or that significant numbers of midges are going undetected because they are biting (and transmitting BTV) during the day when light traps are not effective, and (xi) the sporadic capture of Culicoides in the winter of 2007 invites re-examination of the current definition of a vector-free period. The re-emergence of BT over a wide front in 2007 raises anew questions as to precisely how the virus overwinters and asks also that we scrutinise our monitoring systems in terms of their sensitivity and early warning capability.

Culicoides imicola (Diptera: Ceratopogonidae) is the main vector of the bluetongue virus (btv), the causal agent of bluetongue, in southern Europe. This insect presents mainly an Afro-asian distribution, but it was first reported on mainland Europe in southern Spain ([Mellor and others 1983][1]) and

In August 2006, bluetongue virus (BTV) was detected in the Netherlands, Belgium, western Germany, Luxembourg and northern France for the first time. Consequently, a longitudinal entomological study was conducted in the affected region of northern France (Ardennes) throughout the autumn of 2006. Data on the spatio-temporal distribution of Culicoides (Diptera: Ceratopogonidae) associated with livestock were collected and an attempt was made to identify the vector(s) involved in BTV transmission by means of virus detection in wild-caught biting midges. Weekly sampling using standardized Onderstepoort-type blacklight traps were performed simultaneously both outdoors and indoors in one BTV-free and three BTV-affected farms between September and December 2006. Culicoides were sorted according to farm, location (outdoors vs. indoors), time point (in weeks), species and physiological stage. BTV detection was conducted by RT-PCR on monospecific pools of non-bloodfed parous female Culicoides. The principal results showed: (i) the absence of the Mediterranean vector, C. imicola, (ii) the relatively low abundance of C. dewulfi and C. pulicaris, (iii) the widespread occurrence and abundance of C. obsoletus/C. scoticus with longevity and behaviour compatible with BTV transmission, and (iv) all Culicoides pools tested for BTV were negative. In France, the very low levels of BTV-8 circulation were probably due to the limited introduction of the virus from affected neighbouring countries, and not due to the absence of local vector populations. A key finding has been the substantiation, for the first time, that Culicoides, and particularly the potential vectors C. obsoletus/C. scoticus and C. dewulfi, can be active at night inside livestock buildings and not only outside, as originally believed. The endophagic tendencies of members of the Obsoletus group are discussed in light of the prolonged period of BTV transmission during the autumn of 2006 and the risk of BTV overwintering and resurgence in the spring of 2007. Overall, there is an urgent need to improve our knowledge on the ecology of local Culicoides species before any clear, effective and reliable recommendations can be provided to the veterinary authorities in terms of prevention and control.

Culicoides were captured at a BTV-infected dairy near Gulpen in the province of Limburg (south-east Netherlands) between 14 September and 4 October 2006. Onderstepoort-type blacklight traps were used to sample Culicoides both inside and outside a partially open shed housing 11 cattle. A total of 28 light trap collections were made at the shed and yielded: • 9371 Culicoides representing 11 species; >90% comprised five potential vectors of BTV and in order of abundance were Culicoides obsoletus and Culicoides scoticus (of the Obsoletus Complex), Culicoides dewulfi, Culicoides pulicaris and Culicoides chiopterus; Culicoides imicola, the principal Mediterranean (and African) vector of BTV, was absent. • 2339 Culicoides representing seven species were captured inside (endophily) the cattle shed; >95% comprised the Obsoletus Complex and C. dewulfi. Conversely, the Pulicaris Complex, represented by five species and including C. pulicaris, showed strong exophily with >97% captured outside the shed. • 7032 Culicoides were captured outside the shed, approximately threefold more than inside. This trend was reversed on an overcast day, when eightfold more Culicoides were captured inside; this indicates that when the light intensity outdoors is low Culicoides will attack (i) earlier in the day while cattle are still at pasture, and (ii) might follow cattle into the sheds in the late afternoon leading to elevated numbers of biting midges being trapped inside the shed during the subsequent hours of darkness. • Culicoides were captured inside the shed on all 14 sampling nights. On occasion up to 33% were freshly blood fed indicating they had avidly attacked the cattle inside (endophagy); because half the cattle had seroconverted to BTV, and because no cattle were left outdoors at night, the data indicate that (i) the housing of animals in partially open buildings does not interrupt the transmission of BTV, and/or (ii) BTV is being transmitted while cattle are grazing outdoors during the day. • The capture of partially engorged midges inside the shed shows they are being disturbed while feeding; this may lead to cattle being attacked repeatedly, and if these attacks include older parous BTV-infected Culicoides, may enhance virus dissemination (particularly in sheds where cattle stand close together). • Endo- and exophagy by potential vector Culicoides – coupled to increased adult longevity and multiple feeding events in single (potentially) infected midges – would ensure an R0 of >1, resulting in the continued maintenance and spread of BTV within local vertebrate populations. • Four light trap collections made additionally in a mature deciduous forest 70 m from the shed yielded a high proportion (48%) of gravid females amongst which 10% had incompletely digested blackened blood meals in their abdomens; the absence of this age category in Culicoides captured at the sheds indicates that all Culicoides, after engorgement, exit the buildings to undergo oogenesis elsewhere. In Europe, the blacklight trap is used widely for the nocturnal monitoring of Culicoides; a drawback to this approach is that this trap cannot be used to sample midges that are active during the day. Because diurnal biting in vector Culicoides may constitute a significant and underestimated component of BTV transmission a novel capture methodology will be required in future and is discussed briefly.

International audience; Since 1999, several serotypes of bluetongue virus (BTV) have been isolated in the western part of the Mediterranean basin, and since 2000, Corsica has been exposed to three different serotypes: BTv serotype 2 in 2000, BTv serotype 4 (BTV-4) in 2003 and BTv serotype 16 in 2004. In 2000 there were no surveillance systems for bluetongue, but in 2003, active surveillance of the circulation of BTv and its vector Culicoides species, aided by a raised level of awareness in farmers and veterinarians, made it possible to study the introduction Of BTV-4. The monitoring and analysis of the seroconversions of sentinel herds of goats, clinical signs and meteorological variables showed that the serotype had been present in the island since May that year, but clinical signs were first observed only in October. Moreover, the weather conditions and wind patterns were suitable for the transport of Culicoides species from Sardinia in May. These observations suggest that BTv had been transported on air currents from a southern infected area, and that it could have spread without causing clinical signs of disease for a few months.

SIR, — Zimmer and colleagues ( VR , January 26, 2008, vol 162, p 131) recently described maize silage residues as possible breeding sites for the sibling species Culicoides obsoletus/scoticus and confirmed that Culicoides dewulfi and Culicoides chiopterus breed in cattle dung. To complete the

In southern Europe, orbiviral diseases such as bluetongue (BT) have been assumed to have been largely transmitted by the classical Afro-Asian vector Culicoides imicola Kieffer (Diptera: Ceratopogonidae). Recent outbreaks have occurred in regions where C. imicola is normally absent, supporting the theory that other species belonging to the Obsoletus or Pulicaris complexes may play a role in BT virus transmission. Investigations of the ecology of the species within the former group are hampered by females of member species being extremely difficult to separate by classical morphology. To allow straightforward separation of these species in France, a multiplex polymerase chain reaction-based on internal transcribed spacer (ITS)-1 rDNA was developed to distinguish between Culicoides chiopterus Meigen, Culicoides dewulfi Goetghebuer, Culicoides montanus Shakirjanova, Culicoides obsoletus Meigen, and Culicoides scoticus Downes & Kettle. This tool will be useful in defining both the vector role and larval biotopes of these species in Europe.

The current microscopy method for identifying the Culicoides imicola Kieffer, 1913 species can be time and labour intensive. There is a need for the development of a rapid and quantitative tool to quantify the biting midges C. imicola ss in light trap catches. A reproducible and sensitive real-time polymerase chain reaction method that targets the internal transcribed spacer (ITS-1) of ribosomal DNA of C. imicola ss species was developed. This real-time PCR assay was first performed on 10-fold serial dilutions of purified plasmid DNA containing specific C. imicola ss ITS-1. It was then possible to construct standard curves with a high correlation coefficient (r2 = 0.99) in the range of 10−2–10−8 ng of purified DNA. The performances of this PCR were evaluated in comparison with morphological determination on Culicoides trapped along the Mediterranean coastal mainland France. ROC statistical analysis was carried out using morphology as gold standard and the area under the ROC curve had a satisfactory value of 0.9752. The results indicated that this real-time PCR assay holds promise for monitoring C. imicola ss population in both surveillance and research programmes because of its good specificity (92%) and sensitivity (95%).

International audience; The recent and rapid spread in the Mediterranean Basin of bluetongue, a viral disease of ruminants transmitted by some species of Culicoides (biting midges), highlights the necessity of determining the conditions of its emergence. This study uses high spatial resolution satellite imagery and methods from landscape ecology science to identify environmental parameters related to bluetongue occurrence in Corsica, a French Mediterranean island where the disease occurred for the first time in 2000. A set of environmental variables recorded in the neighborhood of 80 sheep farms were related to case occurrence through a logistic regression model computed within three subsequent buffer distances of 0.5, 1 and 2 km. The results reveal the role of landscape metrics, particularly those characterizing land-use units such as prairies and woodlands, as well as farm type, latitude and sunshine to explain the presence of bluetongue. Internal and external validation both indicate that the best results are obtained with the 1 km buffer size model (area under Receiver Operating Characteristic curve = 0.9 for internal validation and 0.81 for external validation). The results show that high spatial resolution remote sensing (i.e. 10 m pixels) and landscape ecology approaches contribute to improving the understanding of bluetongue epidemiology.

New contribution to the study of the Culicoides of Corsica. Update of the list of species, description of C. riebi n. sp. and redescription of C. paradisionensis Boorman, 1988 (Diptera, Ceratopogonidae). This paper presents the complementary list of Culicoides species collected in Corsica, from October 2001 to December 2003. It gives the description of C. riebi, a new species of biting midge, and a redescription of C. paradisionensis Boorman, 1988, the female of which is described and illustrated for the first time., Cet article présente la liste complémentaire des espèces de Culicoides récoltées en Corse d'octobre 2001 à décembre 2003. Il donne la description de C. riebi n. sp. et la redescription de C. paradisionensis Boorman, 1988, dont la femelle est ici décrite pour la première fois., Delécolle Jean-Claude, Mathieu Bruno, Baldet Thierry. Nouvelle contribution à l'étude des Culicoides de Corse. II. Mise à jour de la liste des espèces, description de C. riebi n. sp. et redescription de C. paradisionensis Boorman, 1988 (Diptera, Ceratopogonidae). In: Bulletin de la Société entomologique de France, volume 110 (1), mars 2005. pp. 69-76.

International audience; Bluetongue (BT) and African Horse Sickness (AHS) are infectious arthropod-borne viral diseases affecting ruminants and horses, respectively. Culicoides imicola Kieffer, 1913, a biting midge, is the principal vector of these livestock diseases in Africa and Europe. Recently bluetongue disease has re-emerged in the Mediterranean Basin and has had a devastating effect on the sheep industry in Italy and on the islands of Sicily, Sardinia, Corsica and the Balearics, but fortunately, has not penetrated onto mainland France and Spain. To survey for the presence of C. imicola, an extensive light-trap network for the collection of Culicoides, was implemented in 2002 in southern mainland France. The morphological identification of Culicoides can be both tedious and time-consuming because its size ranges from 1.5 to 3 mm. Therefore, an ITS1 rDNA polymerase chain reaction (PCR)-based diagnostic assay was developed to rapidly and reliably identify Culicoides spp. and C. imicola. The aim of this work was to set up a rapid test for the detection of C. imicola amongst a pool of insects collected in areas at risk for BT. The sequence similarity of the rDNA (nuclear ribosomal DNA), which is greater within species than between species, is the foundation of its utilisation in species-diagnostic assays. The alignment of the 11 ITS1 sequences of Culicoides obtained from Genbank and EMBL databases helped us to identify one region in the 5' end and one in the 3' end that appear highly conserved. PCR primers were designed within these regions to amplify genus-specific fragments. In order to set up a C. imicola-specific PCR, another forward primer was designed and used in combination with the previously designed reverse primer. These primers proved to be highly specific and sensitive and permitted a rapid diagnostic separation of C. imicola from Culicoides spp.

La fièvre catarrhale ovine (FCO) est une maladie virale qui est transmise par des moucherons piqueurs (Diptera : Ceratopogonidae) et émerge en Méditerranée depuis 1998. Cette émergence est reliée à l’expansion septentrionale de son vecteur principal Culicoides imicola Kieffer 1913. En France, C. imicola a été décelé dans le sud de la Corse en octobre 2000, quelques jours avant l’apparition de foyers de FCO sérotype 2. Un réseau de surveillance entomologique a été mis en place en 2002 : (i) en Corse, pour étudier les dynamiques de populations de C. imicola et des autres espèces de Culicoides associées aux élevages ; (ii) sur le continent, pour détecter l’introduction de C. imicola. Des captures nocturnes ont été réalisées toutes les trois semaines sur 12 sites en Corse et une fois par mois sur 19 sites sur le continent. En 2002, en Corse, un total de 180 nuits de captures ont été effectuées. C. imicola comptait en moyenne pour 18 p. 100 des effectifs totaux capturés et prédominait au sud de l’île. Les densités adultes ont atteint un pic en septembre. La présence de C. imicola sur tous les sites et durant huit mois de l’année a confirmé que le vecteur principal de la FCO passait l’hiver et était installé durablement dans l’île. En outre, en 2003, les populations de C. imicola étaient en pleine expansion dans le Nord, particulièrement en Balagne. Le risque d’invasion du continent devient ainsi plus important. Sur le continent, sur un total de 233 nuits de captures réalisées en 2002-2003, deux spécimens de C. imicola ont été détectés en 2003 sur deux sites différents. Des investigations poussées n’ont pu mettre en évidence des populations locales installées. Les dynamiques saisonnière et spatiale de C. imicola et des autres espèces d’intérêt sont discutées en relation avec leur bioécologie et les facteurs environnementaux. Cette « entomosurveillance » apparaît indispensable pour une meilleure compréhension de l’épidémiologie de la FCO mais aussi pour élaborer et valider des modèles de prédiction de zones à risque.

International audience; Background: African horse sickness (AHS) is an equine disease endemic to Senegal. The African horse sickness virus (AHSV) is transmitted to the mammalian hosts by midges of the Culicoides Latreille genus. During the last epizootic outbreak of AHS in Senegal in 2007, 1,169 horses died from this disease entailing an estimated cost of 1.4 million euros. In spite of the serious animal health and economic implications of AHS, very little is known about determinants involved in transmission such as contact between horses and the Culicoides species suspected of being its vectors. Methods: The monthly variation in host/vector contact was determined in the Niayes area, Senegal, an area which was severely affected by the 2007 outbreak of AHS. A horse-baited trap and two suction light traps (OVI type) were set up at each of five sites for three consecutive nights every month for one year. Results: Of 254,338 Culicoides midges collected 209,543 (82.4%) were female and 44,795 (17.6%) male. Nineteen of the 41 species collected were new distribution records for Senegal. This increased the number of described Culicoides species found in Senegal to 53. Only 19 species, of the 41 species found in light trap, were collected in the horse-baited trap (23,669 specimens) largely dominated by Culicoides oxystoma (22,300 specimens, i.e. 94.2%) followed by Culicoides imicola (482 specimens, i.e. 2.0%) and Culicoides kingi (446 specimens, i.e. 1.9%). Conclusions: Culicoides oxystoma should be considered as a potential vector of AHSV in the Niayes area of Senegal due to its abundance on horses and its role in the transmission of other Culicoides-borne viruses.

Background: Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) are biological vectors of internationally important arboviruses. To understand the role of Culicoides in the transmission of these viruses, it is essential to correctly identify the species involved. Within the western Palaearctic region, the main suspected vector species, C. obsoletus, C. scoticus, C. dewulfi and C. chiopterus, have similar wing patterns, which makes it difficult to separate and identify them correctly. Methods: In this study, designed as an inter-laboratory ring trial with twelve partners from Europe and North Africa, we assess four PCR-based assays which are used routinely to differentiate the four species of Culicoides listed above. The assays based on mitochondrial or ribosomal DNA or microarray hybridisation were tested using aliquots of Culicoides DNA (extracted using commercial kits), crude lysates of ground specimens and whole Culicoides (265 individuals), and non-Culicoides Ceratopogonidae (13 individuals) collected from across Europe. Results: A total of 800 molecular assays were implemented. The in-house assays functioned effectively, although specificity and sensitivity varied according to the molecular marker and DNA extraction method used. The Obsoletus group specificity was overall high (95-99%) while the sensitivity varied greatly (59.6-100%). DNA extraction methods impacted the sensitivity of the assays as well as the type of sample used as template for the DNA extraction. Conclusions: The results are discussed in terms of current use of species diagnostic assays and the future development of molecular tools for the rapid differentiation of cryptic Culicoides species. Published