Thierry Baldet

The 2006 outbreak of bluetongue in northern Europe. The entomological perspective

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.

Assessment of vector/host contact: comparison of animal-baited traps and UV-light/suction trap for collecting Culicoides biting midges (Diptera: Ceratopogonidae), vectors of Orbiviruses

BackgroundThe 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/resultsCollections 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.ConclusionsOnly 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.

Phylogenetic analysis of Culicoides species from France based on nuclear ITS1‐rDNA sequences

Abstract.u2002 Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) play important roles in the transmission of viral diseases affecting wild and domestic ruminants and horses, including Bluetongue (BT) and African horse sickness (AHS) respectively. In southern Europe, BT has been largely transmitted by the classical Afro‐Asian vector Culicoides imicola Kieffer. However, other species such as C.u2003obsoletus Meigen, C.u2003scoticus Downs & Kettle and C.u2003pulicaris Linné may also be involved in BTV transmission. As a consequence of the discovery of C.u2003imicola followed by BTV‐2 outbreaks on the island of Corsica in October 2000, further studies on these biting midges have been carried out. To better characterize the evolution and phylogenetic relations of Culicoides, molecular analysis in parallel with a morphology‐based taxonomic approach were performed. Phylogenetic analyses of French Culicoides species were undertaken using the ribosomal DNA (rDNA) internal transcribed spacer 1 (ITS1) as a molecular target. This region was shown to be useful in understanding evolutionary and genetic relationships between species. Construction of several trees showed that molecular phylogeny within the genus Culicoides correlates not only with morphological‐based taxonomy but also with ecological patterns.

Endophily in Culicoides associated with BTV-infected cattle in the province of Limburg, south-eastern Netherlands, 2006.

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.

Biting midges overwintering in Belgium

SIR, — In Belgium, the first case of blue-tongue was notified on August 18, 2006. Up to March 7, 2007, bluetongue has been diagnosed on 399 sheep and 296 cattle farms ( ) and the last clinical case was recorded on January 15, 2007.nnFollowing the emergence of blue-tongue in Belgium

Using remote sensing to map larval and adult populations of Anopheles hyrcanus (Diptera: Culicidae) a potential malaria vector in Southern France

BackgroundAlthough malaria disappeared from southern France more than 60 years ago, suspicions of recent autochthonous transmission in the French Mediterranean coast support the idea that the area could still be subject to malaria transmission. The main potential vector of malaria in the Camargue area, the largest river delta in southern France, is the mosquito Anopheles hyrcanus (Diptera: Culicidae). In the context of recent climatic and landscape changes, the evaluation of the risk of emergence or re-emergence of such a major disease is of great importance in Europe. When assessing the risk of emergence of vector-borne diseases, it is crucial to be able to characterize the arthropod vectors spatial distribution. Given that remote sensing techniques can describe some of the environmental parameters which drive this distribution, satellite imagery or aerial photographs could be used for vector mapping.ResultsIn this study, we propose a method to map larval and adult populations of An. hyrcanus based on environmental indices derived from high spatial resolution imagery. The analysis of the link between entomological field data on An. hyrcanus larvae and environmental indices (biotopes, distance to the nearest main productive breeding sites of this species i.e., rice fields) led to the definition of a larval index, defined as the probability of observing An. hyrcanus larvae in a given site at least once over a year. Independent accuracy assessments showed a good agreement between observed and predicted values (sensitivity and specificity of the logistic regression model being 0.76 and 0.78, respectively). An adult index was derived from the larval index by averaging the larval index within a buffer around the trap location. This index was highly correlated with observed adult abundance values (Pearson r = 0.97, p < 0.05). This allowed us to generate predictive maps of An. hyrcanus larval and adult populations from the landscape indices.ConclusionThis work shows that it is possible to use high resolution satellite imagery to map malaria vector spatial distribution. It also confirms the potential of remote sensing to help target risk areas, and constitutes a first essential step in assessing the risk of re-emergence of malaria in southern France.

A metapopulation model to simulate West Nile virus circulation in Western Africa, Southern Europe and the Mediterranean basin

In Europe, virological and epidemiological data collected in wild birds and horses suggest that a recurrent circulation of West Nile virus (WNV) could exist in some areas. Whether this circulation is permanent (due to overwintering mechanisms) or not remains unknown. The current conception of WNV epidemiology suggests that it is not: this conception combines an enzootic WNV circulation in tropical Africa with seasonal introductions of the virus in Europe by migratory birds. The objectives of this work were to (i) model this conception of WNV global circulation; and (ii) evaluate whether the model could reproduce data and patterns observed in Europe and Africa in vectors, horses, and birds. The model was calibrated using published seroprevalence data obtained from African (Senegal) and European (Spain) wild birds, and validated using independent, published data: seroprevalence rates in migratory and resident wild birds, minimal infection rates in vectors, as well as seroprevalence and incidence rates in horses. According to this model, overwintering mechanisms are not needed to reproduce the observed data. However, the existence of such mechanisms cannot be ruled out.

Colonization of the Mediterranean Basin by the vector biting midge species #Culicoides imicola#: an old story

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.

Description of the outbreak of bluetongue in Corsica in 2003, and lessons for surveillance

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.

#Culicoides# (Diptera: Ceratopogonidae) midges, the vectors of African horse sickness virus - a host/vector contact study in the Niayes area of Senegal

BackgroundAfrican 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.MethodsThe 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.ResultsOf 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%).ConclusionsCulicoides 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.