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Mosquitoes in the genus Culex are the primary enzootic maintenance and bridge vectors of the North American encephalitides, now including West Nile virus. This review briefly summarizes the biology of three key vector species in western North America, Culex tarsalis, Cx. pipiens complex and Cx. sligmatosoma, focusing on the long history of research done in California. Topics reviewed include population genetic structure, larval ecology, autogeny, mating behavior, host-seeking behavior, host- selection patterns, and overwintering strategies. These attributes collectively have allowed the successful exploitation of anthropogenically altered ecosystems and enabled the role of these species as maintenance and bridge vectors of arboviruses. [ABSTRACT FROM AUTHOR]

The vector competence of Culex tarsalis Coquillett for the BFS1703 strain of western equine encephalomyelitis virus (WEEV) changed significantly as a function of time after infection, mosquito genotype, and infectious virus dose. After ingesting a high virus dose (5 log10 plaque-forming units [PFU]/0.1 ml), females of the susceptible high virus producer (HVP) strain rapidly amplified virus, developed a disseminated infection, and efficiently transmitted WEEV by 4 days postinfection (dpi). The quantity of virus expectorated peaked at 4 dpi (mean 3.4 log10 PFU), and the percentage of females transmitting per os peaked at 7 dpi (80%); both measures of transmission subsequently decreased to low levels throughout the remainder of infected life. HVP females imbibing a low virus dose (3 log10 PFU/0.1 ml) were infected less frequently and took longer to amplify virus to levels recorded for the high virus dose group and did not transmit virus efficiently, thereby indicating midgut infection and escape barriers were dose and time dependent. These data emphasized the importance of elevated avian viremias in Cx. tarsalis vector competence. Females from the WEEV-resistant (WR) strain and two wildtype strains from Kern and Riverside counties were significantly less susceptible to infection at both high and low doses than was the HVP strain. Overall, females with a high virus titer more frequently had a disseminated infection, but there did not seem to be a distinct threshold demarcating this relationship. In marked contrast, all infected females transmitting virus had body titers >4.3 log10 PFU, and most had titers >4.8 log10 PFU. These data indicated that not all females with a disseminated infection transmitted virus because of the presence of one or more salivary gland barriers. [ABSTRACT FROM AUTHOR]

Each transmission season (April-October), mosquito control districts deploy more than 2,000 white leghorn hens as sentinels in California's arbovirus surveillance program. In 2002, Rhode Island red hybrids were used instead, because of insufficient availability of leghorns. Although arboviral seroconversions occurred in both breeds, their attractiveness to host-seeking mosquitoes was not compared. During August and September 2004, Florida Medical Entomology Laboratory traps, each containing 1 hen, were deployed twice weekly at each of 4 fixed locations at a southern California wetland area. The breed at each location was alternated each trap-night to account for intersite variation in the mosquito population. Six chickens of each breed were rotated through the traps to account for interchicken variation in mosquito attractiveness. Although the mosquito counts varied significantly among trap locations, no significant difference was found between the 2 breeds. Sera collected biweekly detected seroconversions to Flavivirus in 4 of 6 red and 1 of 6 white chickens, not a statistically significant difference. Larger sample sizes or experimental infection of different chicken breeds may identify differences in their response to arbovirus infection. [ABSTRACT FROM AUTHOR]

Sentinel chickens are used to detect western equine encephalomyelitis, St. Louis encephalitis, and West Nile virus activity. Flocks that receive high mosquito exposure will be most effective for surveillance purposes. However, mosquito population indices at the flock sites may only provide an indirect measure of potential exposure. Therefore, we developed an indirect enzyme immunoassay to detect chicken antibodies to salivary gland antigens (SGAs) from Culex tarsalis, the primary arbovirus vector in California. Chickens fed upon by Cx. tarsalis developed an antibody response that was proportional to the amount of exposure. Cross-reactivity between sera from Cx. tarsalis-exposed chickens and SGAs from Culex pipiens quinquefasciatus, Culex pipiens pipiens, Ochlerotatus melanimon, and Ochlerotatus sierrensis was likely due to shared SGAs among these species. This serologic assay for mosquito exposure could be used to evaluate the sensitivity of sentinel flocks for detecting arboviral activity. [ABSTRACT FROM AUTHOR]

Eliminating infected female mosquitoes by aerial applications of ultra-low volume adulticides is the intervention strategy currently recommended to interrupt the epidemic transmission of encephalitis viruses, including West Nile. The current research optimized pyrethrin formulations and evaluated their efficacy in the desert environment of the Coachella Valley, Riverside County, California. After seven trials during 2004, a 1:2 by volume mixture of Pyrenone 25-5 in BVA oil optimized particle size, droplet descent to ground level, and kill of sentinel mosquitoes. Three subsequent experiments used 3 aerial applications of the 1:2 Pyrenone 25-5:BVA oil mixture on alternate nights to suppress Culex tarsalis Coquillett host-seeking abundance over a 1-square-mile target area. Mortality patterns among caged sentinel mosquitoes varied among sites and replicate sprays, indicating variable particle dispersion at ground level within the target area. In addition, mortality was observed for sentinels up to 1 mile downwind from the target area, indicating considerable particle drift. Geometric mean abundance of host-seeking Cx. tarsalis females collected at dry ice-baited traps within each of 3 sprayed and 2 unsprayed negative control strata varied similarly over time, indicating that our sprays minimally impacted the target population or that drift combined with other factors led to widespread area control. Experiments during March and June when recruitment rates were minimal showed general area-wide suppression of abundance following spray, whereas an experiment during September when recruitment rates were high from newly flooded marshes failed to prevent an area-wide increase in abundance. Clearly additional research is needed to standardize the efficacy of aerial applications of pyrethrins in hot dry desert environments.

In August 2006, a pyrethrin insecticide synergized with piperonyl butoxide (EverGreen Crop Protection EC 60-6, McLaughlin Gormley King Company, Golden Valley, MN) was sprayed in ultralow volumes over the city of Davis, CA, by the Sacramento-Yolo Mosquito and Vector Control District to control mosquitoes transmitting West Nile virus. Concurrently, we evaluated the impact of the insecticide on nontarget arthropods by 1) comparing mortality of treatment and control groups of sentinel arthropods, and 2) measuring the diversity and abundance of dead arthropods found on treatment and control tarps placed on the ground. We found no effect of spraying on nontarget sentinel species including dragonflies (Sympetrum corruptum), spiders (Argiope aurantia), butterflies (Colias eurytheme), and honeybees (Apis mellifera). In contrast, significantly higher diversity and numbers of nontarget arthropods were found on ground tarps placed in sprayed versus unsprayed areas. All of the dead nontarget species were small-bodied arthropods as opposed to the large-bodied sentinels that were not affected. The mortality of sentinel mosquitoes placed at the same sites as the nontarget sentinels and ground tarps ranged from 0% to 100%. Dead mosquitoes were not found on the ground tarps. We conclude that aerial spraying with pyrethrins had no impact on the large-bodied arthropods placed in the spray zone, but did have a measurable impact on a wide range of small-bodied organisms.

Six experimental ground ultra-low volume (ULV) applications of Pyrenone 25-5 (0.0025 lb/acre) and Aqua-Reslin (0.007 lb/acre) were made by truck-mounted Pro-Mist or London Fog equipment over 1-mi2 study areas in rural and residential environments of the Coachella Valley, Riverside County, California. Efficacy of replicate applications was evaluated by measuring mortality among caged sentinel mosquitoes, by evaluating changes in host-seeking abundance at replicated dry ice-baited traps positioned along intersecting east-west and north-south transects, and by differential recapture patterns of marked females released near traps in the sprayed central core and unsprayed control areas. Sentinel mortality agreed well with estimates of droplet density measured by "slide spinners" and was affected by 1) distance of cages from the truck route; 2) landscape features, such as tree lines that created wind shadows; 3) irregular landscape that disrupted the particle cloud; 4) low wind speed that failed to carry the droplet cloud through the environment; and 5) failure of the droplets to penetrate dead airspace within stands of vegetation. Despite variable sentinel mortality, Culex tarsalis relative abundance in rural landscapes within and around our study areas always declined after ULV applications. Concurrent decreases in abundance at traps within sprayed and adjoining unsprayed areas confounded our estimates of percentage of control using Mulla's formula, which compares abundance in sprayed and unsprayed areas pre- and postspray. ULV applications significantly affected recapture patterns, in that recapture rates within the spray zone usually were significantly less than in the unsprayed zone. Collectively, our data indicated that ground ULV applications measurably reduced Cx. tarsalis abundance in rural areas of the Coachella Valley but that further evaluations may be necessary to validate efficacy on Culex quinquefasciatus populations in residential communities.

Female Culex tarsalis fed heparinized chicken blood-western equine encephalomyelitis virus (WEEV) mixtures through a biomembrane feeder were compared with females fed sweetened blood-virus mixtures presented in pledgets or as hanging drops or to restrained chickens with natural or artificial viremias. Results indicated that sodium heparin did not adversely affect the infection of Culex tarsalis with WEEV. Overall advantages of the biomembrane system included 1) increased blood feeding frequency, 2) control of the infectious virus dose, and 3) greater or similar infection rates and body titers to females taking blood meals from viremic chickens. Anesthetizing females with triethylamine for in vitro transmission assessment using the capillary tube method produced results similar to immobilization using cold or CO2 + cold. Our research provided insight into tools useful to investigate the infection and transmission of WEEV by Cx. tarsalis.