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Commercially available formulations of two entomopathogenic fungi, Beauveria bassiana (Bals.-Criv.) Vuill. (Hypocreales: Clavicipitaceae) and Metarhizium anisopliae (Metchnikoff) Sorokin (Hypocreales: Clavicipitaceae), were assessed for control of Culex quinquefasciatus Say (Diptera: Culicidae) in underground storm drain systems (USDS) in the Coachella Valley of southern California. Each of three treatments, the two fungi or a water control, was applied to 1 m2 of vertical wall at eight USDS sites in spring and autumn of 2015. Fungal infectivity and lethality were assessed at 1 d and 1, 2, and 4 wk post-application. Overnight bioassays using adult lab-reared female mosquitoes were carried out on the treated USDS wall areas and then mosquitoes were held in the laboratory for up to 21 d to allow fungal infections to be expressed. Postmortem fungal sporulation was assessed up to 2 wk at 100% humidity. Mosquito-fungal interactions also were assessed in bioassays of the three treatments on filter paper exposed to USDS conditions during autumn. Metarhizium anisopliae killed mosquitoes faster than B. bassiana; nevertheless, both freshly applied formulations caused greater than 80% mortality. Fungal persistence declined significantly after 1 wk under USDS conditions, but some infectivity persisted for more than 4 wk. Beauveria bassiana was more effective against Cx. qinquefasciatus in the spring, while M. anisopliae was more effective in the cooler conditions during autumn. USDS environmental conditions (e.g., temperature, relative humidity, standing water) influenced fungal-related mortality and infection of Cx. quinquefasciatus. The utility of these fungal formulations for mosquito abatement in the Coachella Valley and implications for fungal control agents in USDS environments are discussed.

Culicoides sonorensis Wirth and Jones (Diptera: Ceratopogonidae) is a key bluetongue virus vector in the United States. Immatures occur in mud near the edges of wastewater ponds and are understudied targets for control efforts. Eggs of C. sonorensis were collected in the morning from a dairy wastewater pond bank by taking 5-ml surface mud samples along four transects on each of six dates. Surface mud samples parallel to waterline (10-cm long, 1-cm wide, and 0.5-cm deep) were removed at 5-cm increments ranging from 15 cm below waterline up to 25 cm above waterline. Eggs were removed using MgSO4 flotation, held on moist filter paper, and scored for hatching over 3 d. Eggs hatching on days 2 and 3 were assumed to have been laid on the test night. Water levels were stable within a night according to time-lapse camera photos. Most samples from below the waterline had no eggs and were not analyzed statistically. Mean (±SE) sample moisture (25.8 ± 2.1 at 5 cm above waterline and 19.8 ± 2.6% at 25 cm above waterline) did not vary significantly by position above waterline. The highest density of viable eggs (21 eggs/5 ml), proportion of mud samples positive for viable eggs (75%), and proportion of eggs hatching (80%) were found 5 cm above waterline. Oviposition in the few hours after sunset is adaptive, allowing eggs to age, develop the serosal cuticle, and resist later desiccation. As a potential control method, reducing water levels after midnight would encourage young egg desiccation.

Laboratory and field research was conducted to determine if Culex tarsalis Coquillett expectorated West Nile virus (WNV) during sugar feeding and if a lure or bait station could be developed to exploit this behavior for WNV surveillance. Experimentally infected Cx. tarsalis repeatedly expectorated WNV onto filter paper strips and into vials with wicks containing sucrose that was readily detectable by a quantitative reverse transcriptase-polymerase chain reaction assay. Few females (33%, n = 27) became infected by imbibing sugar solutions spiked with high concentrations (10(7) plaque forming units/ml) of WNV, indicating sugar feeding stations probably would not be a source of WNV infection. In nature, sugar bait stations scented with the floral attractant phenyl acetaldehyde tracked WNV transmission activity in desert but not urban or agricultural landscapes in California. When deployed in areas of the Coachella Valley with WNV activity during the summer of 2011, 27 of 400 weekly sugar samples (6.8%) tested positive for WNV RNA by reverse transcriptase-polymerase chain reaction. Prevalence of positives varied spatially, but positive sugar stations were detected before concurrent surveillance measures of infection (mosquito pools) or transmission (sentinel chicken seroconversions). In contrast, sugar bait stations deployed in urban settings in Los Angeles or agricultural habits near Bakersfield in Kern County supporting WNV activity produced 1 of 90 and 0 of 60 positive weekly sugar samples, respectively. These results with sugar bait stations will require additional research to enhance bait attractancy and to understand the relationship between positive sugar stations and standard metrics of arbovirus surveillance.