Mosquito species associated with horses in Madagascar: a review of their vector status with regard to the epidemiology of West Nile fever.

In Madagascar, the high West Nile virus (WNV) antibody prevalence reported in horse populations suggests a high level of vector-horse contact. This study aims to characterize the mosquito species usually involved in WNV transmission in horse stables in Madagascar. Five horse stables were investigated in October and November 2016 in five distinct inland areas. Mosquitoes were collected using double net traps baited with human, poultry or horse as well as light traps. Blood meal identification from engorged females was performed using host-specific PCRs. A total of 2898 adult mosquitoes were collected with Culex (Culex) antennatus (Becker) (40.7%), and Cx. (Cux.) quinquefasciatus Say (14.9%), being the most abundant species. The mosquito abundance varied between horse stables (P < 10-7 ) and depending on the bait used in the double net traps (P < 0.003). Among the 190 tested blood meals, 119 consisted of single blood meals with 85 from horse, 17 from human, 16 from chicken, one from cattle and 71 consisted of mixed blood meals. The mosquito species collected during this study exhibited a generalist feeding behaviour allowing them to act as bridge vectors between different vertebrate hosts involved in WNV transmission cycle. Their vector status with regard to West Nile fever epidemiology is reviewed.

Morphometric discrimination of two sympatric sibling species in the Palaearctic region, Culicoides obsoletus Meigen and C. scoticus Downes & Kettle (Diptera: Ceratopogonidae), vectors of bluetongue and Schmallenberg viruses.

BACKGROUND: Some Palaearctic biting midge species (subgenus Avaritia) have been implicated as vectors of bluetongue virus in northern Europe. Separation of two species (C. obsoletus and C. scoticus) is considered difficult morphologically and, often, these female specimens are grouped in entomological studies. However, species-specific identification is desirable to understand their life history characteristics, assess their roles in disease transmission or measure their abundance during arboviral outbreaks. This study aims to investigate whether morphometric identification techniques can be applied to female C. obsoletus and C. scoticus individuals trapped at different geographical regions and time periods during the vector season. METHODS: C. obsoletus and C. scoticus were collected using light-suction traps from the UK, France and Spain, with two geographical locations sampled per country. A total of 759 C. obsoletus/C. scoticus individuals were identified using a molecular assay based on the cytochrome c oxidase subunit I gene. Fifteen morphometric measurements were taken from the head, wings and abdomen of slide-mounted specimens, and ratios calculated between these measurements. Multivariate analyses explored whether a combination of morphometric variables could lead to accurate species identification. Finally, Culicoides spp. collected in France at the start, middle and end of the adult vector season were compared, to determine whether seasonal variation exists in any of the morphometric measurements. RESULTS: The principal component analyses revealed that abdominal characteristics: length and width of the smaller and larger spermathecae, and the length of the chitinous plates and width between them, are the most reliable morphometric characteristics to differentiate between the species. Seasonal variation in the size of each species was observed for head and wing measurements, but not abdominal measurements. Geographical variation in the size of Culicoides spp. was also observed and is likely to be related to temperature at the trapping sites, with smaller individuals trapped at more southern latitudes. CONCLUSIONS: Our results suggest that female C. obsoletus and C. scoticus individuals can be separated under a stereomicroscope using abdominal measurements. Although we show the length and width of the spermathecae can be used to differentiate between the species, this can be time-consuming, so we recommend undertaking this using standardized subsampling of catches.

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.

First overview of the Culicoides Latreille (Diptera: Ceratopogonidae) livestock associated species of Reunion Island, Indian Ocean.

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.

Culicoides vectors of bluetongue virus in Chester Zoo.

On four nights in June 2008, light traps were operated for Culicoides biting midges, the vector species for bluetongue virus (BTV), at five sites in Chester Zoo in north-west England. Over 35,000 Culicoides midges, of 25 species, were captured, including high densities inside animal enclosures. Over 94 per cent of all the Culicoides trapped were females of the Obsoletus group, which is implicated as the vector of BTV serotype 8 in northern Europe. The mean catch of this group per trap per night was over 1500, suggesting a potential risk of BTV transmission if the virus is introduced to Chester Zoo in the animals or midges in the summer.

Modelling local dispersal of bluetongue virus serotype 8 using random walk.

The knowledge of the place where a disease is first introduced and from where it later spreads is a key element for the understanding of an epizootic. For a contagious disease, the main method is back tracing. For a vector-borne disease such as the Bluetongue virus serotype 8 epizootic that occurred in 2006 in North-Western Europe, the efficiency of tracing is limited because many infected animals are not showing clinical signs. In the present study, we propose to use a statistical approach, random walk, to model local spread in order to derive the Area of First Infection (AFI) and spread rate. Local spread is basically described by the random movements of infected insect vectors. Our model localised the AFI centre, origin of the infection, in the Netherlands, South of Maastricht. This location is consistent with the location of the farms where the disease was first notified in the three countries (Netherlands, Belgium, and Germany) and the farm where retrospectively the earliest clinical signs were found. The derived rate of spread of 10-15 km/week is consistent with the rates observed in other Bluetongue epizootics. In another article Mintiens (2008), the AFI definition has then been used to investigate possible ways of introduction (upstream tracing) and to study the effect of animal movements from this area (downstream tracing).

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.

Clusters of autochthonous hepatitis A cases in a low endemicity area.

At the University Hospital of Besançon (département of Doubs, France), an unusually high number of patients were hospitalized for hepatitis A during the 1999-2000 period, some of whom had not travelled abroad. This prompted us to conduct an investigation on a population basis and search for clusters of cases possibly related to local sources of contamination. Accordingly, case definition was restricted to autochthonous cases. During the 1999-2002 period, 45 autochthonous cases were classified as possibly originating from local environmental sources. A space-time scan statistic detected one most likely cluster (standardized incidence ratio 20.63, 95% confidence interval 10.6-37.1), consisting of 11 persons (of whom five children had attended the same swimming pool). It remained significant in a sensitivity analysis, strongly supporting the hypothesis of an environmental source of contamination. This study reveals the necessity of regular surveillance for hepatitis A and raises the issue of virological surveys of pool waters.