World Health Organisation test applied on Culicoides: a multicentric study

The aim of this work is to organize a multicentric assay to compare susceptibilities of Culicoides populations to insecticides (i.e. C. imicola and C. obsoletus sensu stricto). The base line of the World Health Organisation test kit (WHO/VBC81.805) is to expose mosquitoes to papers impregnated with insecticides to detect the resistance of Anopheles (paludism vector) in the field. In a novel approach, the WHO test can be used on Culicoides to assess midges' susceptibility to insecticides. Information on the intrinsic susceptibilities to insecticides is required to assess correctly the existing (or new) formulations of insecticide. This multicentric assay will improve our capacity to experiment by networking and assess the population variability by comparing susceptibilities between different populations of Culicoides. Field-collected midges are trapped alive using an ultraviolet light trap (OVI model, South Africa) and replacing the collection jar with a fine mesh cage. To prevent desiccation, cages are covered with wet papers, wrapped in aluminium foil and retrieved at dawn. It is important to choose the site and period with abundant target species to improve robustness with more individuals. Experiments should be carried out in an insecticide-free environment with constant temperature and humidity, and started a few hours after collection. Insecticide-impregnated papers are rolled and fastened into the WHO kit test tube. About 30 Culicoides (approximately 20 nulliparous females) are collected and transferred to the tube. Tubes are maintained horizontally for an hour (exposure to insecticide). Mortalities are recorded 1 h and 24 h after exposure. One control is performed without insecticide to validate the test. Mortalities are analysed by probit regression to obtain LC50 and LC90 for each population. The test has to be run at least with four concentrations of insecticide, one control and four replicates with samples from the same population. Only nulliparous females are recorded. Insecticide concentrations are given in percentages (WHO standard). In this WHO test, the target species are C. imicola and C. obsoletus s.s., and the active ingredient used is deltamethrin at doses 0.005, 0.001, 0.0005 and 0.0001%. Data will be centralized and analysed by EID-Méditerranée (France), and results will be given back to the partners and published. Future studies could focus on comparing the four deltamethrin concentrations on other species and testing Culicoides susceptibility to other components. (Texte intégral

Spatio-temporal genetic variation of the biting midge vector species Culicoides imicola (Ceratopogonidae) Kieffer in France

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. (Résumé d'auteur