Attractiveness of a four-component pheromone blend to male navel orangeworm moths.

The attractiveness to male navel orangeworm moth, Amyelois transitella, of various combinations of a four-component pheromone blend was measured in wind-tunnel bioassays. Upwind flight along the pheromone plume and landing on the odor source required the simultaneous presence of two components, (11Z,13Z)-hexadecadienal and (3Z,6Z,9Z,12Z,15Z)-tricosapentaene, and the addition of either (11Z,13Z)-hexadecadien-1-ol or (11Z,13E)-hexadecadien-1-ol. A mixture of all four components produced the highest levels of rapid source location and source contact. In wind-tunnel assays, males did not seem to distinguish among a wide range of ratios of any of the three components added to (11Z,13Z)-hexadecadienal. Dosages of 10 and 100 ng of the 4-component blend produced higher levels of source location than dosages of 1 and 1,000 ng.

Temporal changes in distribution, prevalence and intensity of northern fowl mite (Ornithonyssus sylviarum) parasitism in commercial caged laying hens, with a comprehensive economic analysis of parasite impact.

Establishment and spread of Ornithonyssus sylviarum were documented through time on sentinel hens (50 per house of 28,000-30,000 hens) in the first egg production cycle of three large commercial flocks (12 houses) of white leghorn hens. Mites were controlled using acaricide, and the impacts of treatment on mite populations and economic performance were documented. Mite prevalence and intensity increased rapidly and in tandem for 4-8 weeks after infestation. Intensity declined due to immune system involvement, but prevalence remained high, and this would affect mite sampling plan use and development. Early treatment was more effective at controlling mites; 85% of light infestations were eliminated by a pesticide spray (Ravap), versus 24% of heavy infestations. Hens infested later developed lower peak mite intensities, and those mite populations declined more quickly than on hens infested earlier in life. Raw spatial association by distance indices (SADIE), incorporating both the intensity and distribution of mites within a house, were high from week-to-week within a hen house. Once adjusted spatially to reflect variable hen cohorts becoming infested asynchronously, this analysis showed the association index tended to rebound at intervals of 5-6 weeks after the hen immune system first suppressed them. Large, consistent mite differences in one flock (high vs. low infestation levels) showed the economic damage of mite parasitism (assessed by flock indexing) was very high in the initial stages of mite expansion. Unmitigated infestations overall reduced egg production (2.1-4.0%), individual egg weights (0.5-2.2%), and feed conversion efficiency (5.7%), causing a profit reduction of $0.07-0.10 per hen for a 10-week period. Asynchronous infestation patterns among pesticide-treated hens may have contributed to a lack of apparent flock-level economic effects later in the production cycle. Individual egg weights differed with mite loads periodically, but could be either higher or lower, depending on circumstances and interactions with hen weight. Individual hen weight gains were depressed by high/moderate mite loads, but the heavier hens in a flock harbored more mites. This led to compensatory weight gains after mites declined. Tradeoffs between resource allocation to body growth or production versus immune system function appeared to be operating during the early and most damaging mite infestation period, when high egg production was beginning and the hens were gaining weight. The results were related to other studies of mite impact on domestic hens and to wild bird-ectoparasite studies. Much of the mite economic damage probably is due to engaging and maintaining the immune response.

Growth characteristics of ChimeriVax-Den vaccine viruses in Aedes aegypti and Aedes albopictus from Thailand.

Four chimeric yellow fever (YF) 17D-dengue (DEN) candidate vaccine viruses (ChimeriVax-DEN; Acambis, Cambridge, MA) were characterized in Aedes aegypti and Ae. albopictus mosquitoes collected from Thailand. The four vaccine viruses contained the relevant prM and E genes of wild-type dengue viruses (DENV; serotypes 1-4) substituted for the equivalent genes in the YF vaccine virus (17D) backbone. Each chimera conferred protection against the homologous DENV serotype; a tetravalent mix of all four chimeras stimulates an immune response against all serotypes. Field-collected mosquitoes from Thailand were fed on blood containing each of the viruses under study and held 21 days after infection. Infection and dissemination rates were based on antigen detection in the body or head tissues, respectively. All four wild-type DENV serotypes infected and disseminated, but the candidate vaccine viruses were highly attenuated in mosquitoes with respect to infection and especially with respect to dissemination. Considering the low level viremias anticipated in humans vaccinated with these viruses, it is predicted that the risks of infection and transmission by mosquitoes in nature is minimal.

Determinants of vector specificity of o'nyong nyong and chikungunya viruses in Anopheles and Aedes mosquitoes.

The alphaviruses o'nyong nyong virus (ONNV) and chikungunya virus (CHIKV) provide a unique system to study the viral genes involved in vector specificity. ONNV infects both anopheline and culicine mosquitoes, whereas CHIKV infects only culicine mosquitoes. In this study, chimeric viruses were constructed that contained genes from both ONNV and CHIKV. These chimeras and previously described full-length infectious clones of ONNV and CHIKV were evaluated in Anopheles gambiae and Aedes aegypti mosquitoes. Virus derived from the infectious clones of ONNV and CHIKV retained the vector specificity of the parental viruses. All six of the chimeras were found to infect Ae. aegypti mosquitoes at high rates but only the chimera containing viral genes encoding all of the structural proteins of ONNV was able to infect An. gambiae mosquitoes. These data indicate that all of the viral structural proteins are necessary for ONNV to infect An. gambiae mosquitoes.

West Nile virus dissemination and tissue tropisms in orally infected Culex pipiens quinquefasciatus.

We investigated the spatial and temporal distribution of West Nile virus (WNV) in organs and tissues of Culex pipiens quinquefasciatus mosquitoes for up to 27 days following oral infection. WNV antigen was detected in paraffin-embedded mosquitoes by immunohistochemistry. Immunofluorescence assays were performed on dissected salivary glands and midguts and analyzed by confocal microscopy. We evaluated the route of virus dissemination following midgut escape and the relative importance of amplifying tissues in mosquito susceptibility to infection. WNV infection was persistent in all tissues analyzed including the midgut, salivary glands, nervous system, and fat body and only declined in the cytoplasm of posterior midgut epithelial cells over time. The phenomenon of cell-to-cell spread was observed in the midgut epithelium and WNV intensely infected both circular and longitudinal muscles of the same organ. It is possible that muscle tissue serves as a conduit for virus dissemination and contributes to WNV amplification, particularly late in infection. These findings provide insight into WNV infection dynamics in a highly susceptible, epidemiologically important, North American vector. Our results pave the way for future studies to analyze physical and biological barriers to WNV dissemination in less competent mosquitoes.

Short report: comparison of oral infectious dose of West Nile virus isolates representing three distinct genotypes in Culex quinquefasciatus.

Phylogenetic analysis of West Nile virus in North America has identified replacement of the originally introduced clade, Eastern United States (NY99), by the North American clade. In addition, the transient emergence of other clades and genetic variants has also been observed. In this study, we investigated the potential role of the mosquito in the selection of these clades and genetic variants. We determined the relative susceptibility of Culex quinquefasciatus to infection with isolates from the Eastern U.S. clade, the North American clade, and the Southeast coastal Texas clade. Although significant differences were observed in 50% oral infectious dose values between the Eastern U.S. and two attenuated North American genetic variants compared with the North American and Southeast coastal Texas clade viruses, these differences did not correlate with persistence of the genotype in nature. These results indicate that selection of these viral genotypes was independent of viral oral infectivity in the mosquito.

Nonviremic transmission of West Nile virus.

West Nile virus (WNV) is now the predominant circulating arthropod-borne virus in the United States with >15,000 human cases and >600 fatalities since 1999. Conventionally, mosquitoes become infected when feeding on viremic birds and subsequently transmit the virus to susceptible hosts. Here, we demonstrate nonviremic transmission of WNV between cofeeding mosquitoes. Donor, Culex pipiens quinquefasciatus mosquitoes infected with WNV were fed simultaneously with uninfected "recipient" mosquitoes on naïve mice. At all times, donor and recipient mosquitoes were housed in separate sealed containers, precluding the possibility of mixing. Recipients became infected in all five trials, with infection rates as high as 5.8% and no detectable viremia in the hosts. Remarkably, a 2.3% infection rate was observed when 87 uninfected mosquitoes fed adjacent to a single infected mosquito. This phenomenon could potentially enhance virus survival, transmission, and dispersion and obviate the requirement for viremia. All vertebrates, including immune and insusceptible animals, might therefore facilitate mosquito infection. Our findings question the status of dead-end hosts in the WNV transmission cycle and may partly explain the success with which WNV established and rapidly dispersed throughout North America.