Culicoides midge bites modulate the host response and impact on bluetongue virus infection in sheep.

Many haematophagous insects produce factors that help their blood meal and coincidently favor pathogen transmission. However nothing is known about the ability of Culicoides midges to interfere with the infectivity of the viruses they transmit. Among these, Bluetongue Virus (BTV) induces a hemorrhagic fever- type disease and its recent emergence in Europe had a major economical impact. We observed that needle inoculation of BTV8 in the site of uninfected C. nubeculosus feeding reduced viraemia and clinical disease intensity compared to plain needle inoculation. The sheep that developed the highest local inflammatory reaction had the lowest viral load, suggesting that the inflammatory response to midge bites may participate in the individual sensitivity to BTV viraemia development. Conversely compared to needle inoculation, inoculation of BTV8 by infected C. nubeculosus bites promoted viraemia and clinical symptom expression, in association with delayed IFN- induced gene expression and retarded neutralizing antibody responses. The effects of uninfected and infected midge bites on BTV viraemia and on the host response indicate that BTV transmission by infected midges is the most reliable experimental method to study the physio-pathological events relevant to a natural infection and to pertinent vaccine evaluation in the target species. It also leads the way to identify the promoting viral infectivity factors of infected Culicoides in order to possibly develop new control strategies against BTV and other Culicoides transmitted viruses.

Culicoides species associated with sheep in the Netherlands and the effect of a permethrin insecticide.

Culicoides spp. act as vectors for a number of viral diseases of animals including bluetongue in sheep. The aims of this study were to determine: (1) which Culicoides spp. are associated with sheep in The Netherlands; (2) the time of the day when they are most active; and (3) the effect of treatment of animals with a permethrin insecticide. Two pairs of sheep were each housed within mosquito tents of either one or two layers of netting and all trapped Culicoides spp. were identified microscopically. For the permethrin insecticide study, one of two pairs of sheep was treated with 3.6% permethrin and all animals were housed in tents of similar design. Of the 6210 midges captured, 54.1% were identified as C. chiopterus and 42.7% as C. obsoletus. C. imicola was not identified. The average insect feeding rate was 35-40% and midge activity was greatest around sunset. Permethrin treatment reduced the number of midges captured by 50% and also resulted in a decrease in the percentage of midges that had fed. The findings provide useful information on the behaviour and distribution of Culicoides spp. that will facilitate the development of appropriate control strategies to minimise the risk of insect-vector borne virus diseases such as bluetongue.

Susceptibility of Culicoides biting midge larvae to the insect-pathogenic fungus, Metarhizium anisopliae: prospects for bluetongue vector control.

Culicoides biting midges (Diptera: Ceratopogonidae) are responsible for the spread of several arboviruses of livestock and humans that are of international importance. This study assesses the virulence of 18 insect-pathogenic fungal strains from the genera Metarhizium, Beauveria, Isaria and Lecanicillium to larval stages of Culicoides nubeculous Meigen as a means of examining their potential as biocontrol agents. In initial screening, six strains of M. anisopliae (ERL700, CA1, V275, LRC181A, ARSEF 3291 and ARSEF 4556) outperformed the other tested genera and were found to cause between 90% and 100% larval mortality in all larval instars of this species at 72 h post inoculation. The virulence of the most effective strain, M. anisopliae V275, was then further tested by exposing larvae to doses which ranged from 10(4)-10(8) conidia/ml and recording mortality at 24, 48 and 72 h in a 24-multi-well plate with each well containing 600 microl of water and at 24 and 48 h in 250 ml plastic cups containing 50 ml of water. Sensitivity of larvae was extremely high in the multi-well plates, with LC(50) values of 4.3-4.5 x 10(3)conidia/ml and no significant differences between larval instars. In the 250 ml cups, M. anisopliae V275 caused mortalities of between 70% and 100% to larvae and later instars exhibited higher mortality rates. The results are discussed in relation to incorporation of M. anisopliae into biocontrol programmes to control arboviruses vectored by Culicoides.