Identification of Chemicals Associated Gambusia affinis (Cyprinodontiformes: Poeciliidae), and Their Effect on Oviposition Behavior of Culex tarsalis (Diptera: Culicidae) in the Laboratory.

The western mosquitofish, Gambusia affinis (Baird & Girard), has been used worldwide for the control of larval mosquitoes for more than 100 yr. We found that the western encephalitis mosquito, Culex tarsalis Coquillett (Diptera: Culicidae), can detect the presence of G. affinis in oviposition sites based on associated chemicals, leading to a decrease in the number of egg rafts laid. Three volatile chemical compounds were identified in the headspace above the water where G. affinis had been held for 24 h. Oviposition bioassays conducted using standards of the volatile compounds identified (dimethyl disulfide [DMDS], dimethyl trisulfide [DMTS], and S-methyl methanethiosulphonate) found that females reduced oviposition only when low concentrations of DMTS were present, but this response was not consistent across all trials and concentrations tested. DMDS, DMTS, and S-methyl methanethiosulphonate are known bacterial metabolic waste products and may be the source of the compounds. Two nonvolatile compounds of interest were found to be present in the Gambusia-exudate water. After tasting Cx. tarsalis were deterred from ovipositing onto Gambusia-treated water from which the bacteria had been removed by filtration, indicating that the kairomone may consist of nonvolatile compound(s). One of the nonvolatile compounds isolated from the Gambusia-treated water has a benzene ring structure similar to that of cholesterol but the structure of the two nonvolatile deterrents remains to be fully characterized. Our research shows that three volatile compounds and two nonvolatile compounds are present in water associated with G. affinis (Poeciliidae: Gambusia) and affect the oviposition behavior of Cx. tarsalis in laboratory bioassays.

Evaluation of Two Management Strategies for Harvested Emergent Vegetation on Immature Mosquito Abundance and Water Quality.

Emergent macrophytes play critical roles in water treatment processes of free-water surface constructed treatment wetlands. Management strategies for plant biomass affect wetland function and mosquito populations. Sinking of harvested macrophyte biomass is thought to provide organic carbon that enhances denitrifying bacteria important for nutrient removal while concomitantly reducing harborage for mosquitoes. The effects of sinking versus floating dried plant biomass (California bulrush [Schoenoplectus californicus]) on immature mosquito abundance and water quality (nutrient levels, oxygen demand, and physicochemical variables) were examined in mesocosms (28-m2 ponds or 1.4-m2 wading pools) under different flow regimes in 4 studies. The numbers of mosquito larvae in earthen ponds with floating vegetation were greater than in ponds with sunken vegetation on most dates but did not differ significantly between the 2 vegetation treatments in experiments using wading pools. Differences of the abundance of Anopheles larvae between the 2 vegetation management treatments were larger than for Culex larvae when naturally occurring larval mosquito predators were present. At high turnover rates (>2 pond volumes/day), water quality did not differ significantly between the vegetation management treatments and the water supply. At low turnover rates (approximately 2-6% of water volume/day), water quality differed significantly between the 2 vegetation management treatments and the water supply. Sinking vegetation can enhance the effectiveness of mosquito control but, depending on water management practices, may raise the concentrations of water quality constituents in discharges that are regulated under the Clean Water Act.

Oviposition Behavior of Culex tarsalis (Diptera: Culicidae) Responding to Semiochemicals Associated with the Western Mosquitofish, Gambusia affinis (Cyprinodontiformes: Poecilliidae).

Considerable previous research has focused on predator-associated semiochemicals and how they affect mosquito oviposition behavior. However, most of this work has been done without taking into consideration either the natural density of the predators or how other semiochemicals in aquatic environments might affect the responses of gravid mosquitoes. The influence of mosquitofish density, source water (tap vs pond), presence of freshly laid egg rafts, and removal of a putative source of semiochemicals (bacteria) on oviposition by Culex tarsalis Coquillett (Diptera: Culicidae) was studied in laboratory bioassays. Culex tarsalis females were deterred from laying egg rafts on water that contained semiochemicals associated with Gambusia affinis (Baird & Girard), but this deterrence was not strongly associated with the density of fish used to condition aged tap water. The number of egg rafts laid onto Gambusia-exudate water made with either tap water (density < 1 fish per liter) or pond water was typically ≥ 50% of that onto water that did not house mosquitofish. Gravid mosquitoes tested individually did not reduce oviposition onto Gambusia-exudate water as compared to controls. Likewise, oviposition by females with ablated wings did not differ significantly between Gambusia-exudate water and controls. Oviposition onto filter-sterilized Gambusia-exudate water was reduced relative to unfiltered water, suggesting that semiochemicals deterring egg-laying were still present after bacteria were removed. Taken together, these findings suggest that the responses of gravid Cx. tarsalis to chemicals from habitats containing mosquitofish are complex and the origin of the semiochemicals present in the Gambusia-exudate water needs to be elucidated.

Evaluation of Two Entomopathogenic Fungi for Control of Culex quinquefasciatus (Diptera: Culicidae) in Underground Storm Drains in the Coachella Valley, California, United States.

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.

Effects of a larval mosquito biopesticide and Culex larvae on a freshwater nanophytoplankton (Selenastrum capricornatum) under axenic conditions.

The effects of microbial biopesticides used for mosquito control on autotrophic microorganisms such as nanophytoplankton are equivocal. We examined impacts of mosquito biopesticides and mosquito larvae on primary producers in two independent experiments. In the first experiment, we examined the effects of a commonly used microbial biopesticide formulation (VectoMax® CG) on a unicellular microalga, Selenastrum capricornatum Printz, under axenic laboratory conditions. The biopesticide treatments included two concentrations (0.008 and 0.016 g liter-1 ) of VectoMax® CG and two controls (one untreated and another with autoclaved 0.016 g VectoMax® CG liter-1 ) in replicated axenic experimental microcosms. Spectrophotometric analysis of chlorophyll a (proxy for algal biomass) and direct enumeration of algal cells following the treatments revealed no significant effects of the microbial biopesticide on algal population growth during the four-week study. In the second experiment, we tested the effects of different densities of Culex larvae on the population of S. capricornatum. Effects of mosquito larvae feeding on S. capricornatum were significant with a curvilinear relationship between larval density and algal abundance in the water column. Together, these studies demonstrated a lack of direct cytological/toxicological effects of Bacillus-based microbial pesticides on freshwater primary production and support the hypothesis that the reduction in algal primary production previously reported when Bti products were applied to aquatic environments was likely independent of the Bacillus-based larvicidal toxins. Instead, it was likely mediated by microbial interactions in the water column and the trophic cascade effects that resulted from the removal of larval mosquitoes. These studies suggest that mosquito larvae independent of pesticide application can influence primary production. Our method of evaluating biopesticides against small photoautotrophs can be very useful for studying the unintended effects on autotrophic microorganisms of other pesticides, including herbicides and pesticides applied to aquatic environments.

Large-Volume Gravid Traps Enhance Collection of Culex Vectors.

Gravid mosquito collections were compared among several large-volume (infusion volume ≥35 liters) gravid trap designs and the small-volume (infusion volume  =  6 liters) Centers for Disease Control and Prevention (CDC) gravid trap used routinely by vector control districts for vector and pathogen surveillance. The numbers of gravid Culex quinquefasciatus, Cx. tarsalis, and Cx. stigmatosoma collected by large gravid traps were greater than by the CDC gravid trap during nearly all overnight trials. Large-volume gravid traps collected on average 6.6-fold more adult female Culex mosquitoes compared to small-volume CDC gravid traps across 3 seasons during the 3 years of the studies. The differences in gravid mosquito collections between large-versus small-volume gravid traps were greatest during spring, when 8- to 56-fold more Culex individuals were collected using large-volume gravid traps. The proportion of gravid females in collections did not differ appreciably among the more effective trap designs tested. Important determinants of gravid trap performance were infusion container size and type as well as infusion volume, which determined the distance between the suction trap and the infusion surface. Of lesser importance for gravid trap performance were the number of suction traps, method of suction trap mounting, and infusion concentration. Fermentation of infusions between 1 and 4 wk weakly affected total mosquito collections, with Cx. stigmatosoma collections moderately enhanced by comparatively young and organically enriched infusions. A suction trap mounted above 100 liters of organic infusion housed in a 121-liter black plastic container collected the most gravid mosquitoes over the greatest range of experimental conditions, and a 35-liter infusion with side-mounted suction traps was a promising lesser-volume alternative design.

Oviposition of Culex tarsalis (Diptera: Culicidae) Differs on Water Conditioned by Potential Fish and Insect Predators.

The response of egg-laying Culex tarsalis Coquillett (Diptera: Culicidae) to water conditioned by three fish species used for mosquito control and three predatory aquatic insect species was examined in laboratory binary choice experiments. Oviposition by Cx. tarsalis was 72% less on water conditioned with the arroyo chub, Gila orcutti (Eigenmann & Eigenmann) (Cypriniformes: Cyprinidae) relative to control cups containing aged tap water, but no significant difference was found in the numbers of egg rafts laid on water conditioned with the fathead minnow (Pimephales promelas (Rafinesque), Cypriniformes: Cyprinidae) and the control treatment (water aged 24 h). Mosquito oviposition on water conditioned with the predominantly herbivorous/algivorous California Mozambique tilapia hybrid (Oreochromis mossambicus (Peters) × Oreochromis urolepis hornorum L. (Perciformes: Cichlidae)) or predatory insects (nymphs: Sympetrum corruptum (Hagen) (Odonata: Libellulidae); adults: Thermonectus basillaris (Harris) or Cybister fimbriolatus (Say) (Coleoptera: Dytiscidae)) did not differ significantly relative to that onto water aged for 24 h. As compared with water aged 24 h and water conditioned with diving beetles, oviposition by Cx. tarsalis was significantly lower (≥53%) when live predatory diving beetles were present in oviposition cups. Gravid Cx. tarsalis females do not respond equally to putative semiochemicals in water conditioned with the piscine or aquatic insect predators of immature mosquitoes tested here.

Suitability of monotypic and mixed diets for Anopheles hermsi larval development.

The developmental time and survival to eclosion of Anopheles hermsi Barr & Guptavanij fed monotypic and mixed diets of ten food types were examined in laboratory studies. Larvae fed monotypic diets containing animal detritus (freeze-dried rotifers, freeze-dried Daphnia pulicaria, and TetraMin® fish food flakes) and the mixotrophic protistan Cryptomonas ovata developed faster and survived better than larvae that were fed other monotypic diets. Survival to adulthood of larvae fed several concentrations of the diatom Planothidium (=Achnanthes) lanceolatum was poor (<13%) and larval development time was approximately twice that of larvae fed TetraMin® fish food flakes, the standard laboratory diet. Larvae fed monotypic diets containing prokaryotes (bacteria [Bacillus cereus] and cyanobacteria [Oscillatoria prolifera]) and brewer's yeast (Saccharomyces cerevisiae) failed to survive beyond the 1(st) and 2(nd) instar, respectively. Larvae fed only chlorophytes, single-celled Chlamydomonas reinhardtii and filamentous Spirogyra communis, failed to complete larval development, regardless of the concentration tested. Cohorts fed a combination of food types (mixed diets) usually developed better than cohorts fed monotypic diets. Food types that failed to support complete development when fed alone often facilitated development to adulthood when fed in combination with food types containing >1% C20 polyunsaturated fatty acids as total fat, but regardless of essential fatty acid content, algae that produced mucilage and filaments that sank out of the feeding zone were poor quality diets.

Culex quinquefasciatus larval microbiomes vary with instar and exposure to common wastewater contaminants.

Like many insects, mosquitoes, rely on endosymbionts to grow and develop. These can be acquired from the environment. We used next generation 454 pyrosequencing to discern the whole-body microbiome of the mosquito species Culex quinquefasciatus in various larval stadia and following exposure to common pharmaceutical and personal care products (PPCPs) found in wastewater. PPCP treatments included environmentally-relevant concentrations; 1) a combination of common antibiotics, 2) a combination of mammalian hormones, 3) a mixture of the antibiotic and hormone treatments plus acetaminophen and caffeine and, 4) an untreated control. Within control groups, the predominant families of bacterial symbionts change with each larval instar despite consistent diets and rearing conditions. This trend was also seen in hormone treatments but not in the antibiotic or the mixture treatments. Richness and evenness were reduced in both antibiotic and mixture treatments, suggesting that antibiotics remove certain bacteria or inhibit them from increasing to proportions seen in the control treatment. Interestingly, the mixture treatments had greater richness and evenness compared to antibiotic alone treatments, possibly due to the other contaminants facilitating growth of different bacteria. These findings illuminate the complexity of the microbiome of C. quinquefasciatus and may have implications for more effective control strategies.

Evolution of Resistance in Culex quinquefasciatus (Say) Selected With a Recombinant Bacillus thuringiensis Strain-Producing Cyt1Aa and Cry11Ba, and the Binary Toxin, Bin, From Lysinibacillus sphaericus.

Fourth instars of Culex quinquefasciatus (Say) (Diptera: Culicidae) were selected with a recombinant bacterial strain synthesizing the mosquitocidal proteins from Lysinibacillus sphaericus (Bin) and Cry11Ba and Cyt1Aa from Bacillus thuringiensis. Selection was initiated in Generation 1 with a concentration of 0.04 µg/ml, which rose to a maximum selection concentration of 8.0 µg/ml in Generation 14, followed by an unexpected, rapid increase in mortality in Generation 15. Subsequently, a selection concentration of 0.8 µg/ml was determined to be survivable. During this same period, resistance rose to nearly 1,000-fold (by Generation 12) and declined to 18.8-fold in Generation 19. Resistance remained low and fluctuated between 5.3 and 7.3 up to Generation 66. The cross-resistance patterns and interactions among the component proteins were analyzed to identify possible causes of this unusual pattern of evolution. Poor activity in the mid-range concentrations and lower-than-expected synergistic interactions were identified as potential sources of the early resistance. These findings should be considered in the development of genetically engineered strains intended to control nuisance and vector mosquitoes.

Developmental succession of the microbiome of Culex mosquitoes.

BACKGROUND: The native microflora associated with mosquitoes have important roles in mosquito development and vector competence. Sequencing of bacterial V3 region from 16S rRNA genes across the developmental stages of Culex mosquitoes (early and late larval instars, pupae and adults) was used to test the hypothesis that bacteria found in the larval stage of Culex are transstadially transmitted to the adult stage, and to compare the microbiomes of field-collected versus laboratory-reared mosquitoes. RESULTS: Beta diversity analysis revealed that bacterial community structure differed among three life stages (larvae, pupae and adults) of Culex tarsalis. Although only ~2% of the total number of bacterial OTUs were found in all stages, sequences from these OTUs accounted for nearly 82% of the total bacterial sequences recovered from all stages. Thorsellia (Gammaproteobacteria) was the most abundant bacterial taxon found across all developmental stages of field-collected Culex mosquitoes, but was rare in mosquitoes from laboratory-reared colonies. The proportion of Thorsellia sequences in the microbiomes of mosquito life stages varied ontogenetically with the greatest proportions recovered from the pupae of C. tarsalis and the lowest from newly emerged adults. The microbiome of field-collected late instar larvae was not influenced significantly by differences in the microbiota of the habitat due to habitat age or biopesticide treatments. The microbiome diversity was the greatest in the early instar larvae and the lowest in laboratory-reared mosquitoes. CONCLUSIONS: Bacterial communities in early instar C. tarsalis larvae were significantly more diverse when compared to late instar larvae, pupae and newly emerged adults. Some of the bacterial OTUs found in the early instar larvae were also found across developmental stages. Thorsellia dominated the bacterial communities in field-collected immature stages but occurred at much lower relative abundance in adults. Differences in microbiota observed in larval habitats did not influence bacterial community profiles of late instar larvae or adults. However, bacterial communities in laboratory-reared C. tarsalis larvae differed significantly from the field. Determining the role of Thorsellia in mosquitoes and its distribution across different species of mosquitoes warrants further investigation.

Pharmaceuticals and personal care products alter the holobiome and development of a medically important mosquito.

The increasing demand for fresh water has forced many countries to use reclaimed wastewater for agricultural purposes. This water contains pharmaceuticals and personal care products (PPCPs) that remain biologically active following passage through wastewater treatment plants. Run-off from farms and contaminated water from treatment facilities exposes aquatic ecosystems to PPCPs. This study examined the effects of PPCPs on a lower trophic organism. Culex quinquefasciatus larvae were reared in water contaminated with environmentally relevant concentrations of common PPCPs. Acetaminophen alone and a mixture of contaminants were found to increase developmental time of larvae. Susceptibility to Bti increased in larvae exposed to antibiotics, acetaminophen, or a mixture of PPCPs. Antibiotics, hormones, and the mixture altered the mosquito bacterial microbiome. Overall, the results indicate that at environmentally relevant concentrations, PPCPs in reclaimed water can have biologically important effects on an ecologically and medically important lower trophic level insect.

Effects of nutrients on mosquitoes and an emergent macrophyte, Schoenoplectus maritimus, for use in treatment wetlands.

Schoenoplectus maritimus (alkali bulrush) has desirable attributes, such as a short growth habit (height of mature stands < 1.5 m) and annual senescence, for a potential alternative to tall (height > 3 m) emergent macrophytes in shallow constructed treatment wetlands treating ammonium-dominated wastewater. The effects of different ammonium nitrogen (NH4-N) levels on alkali bulrush growth and its ability to take up nutrients from the wastewater, as well as on mosquito production, across the range of NH4-N found in constructed wetlands of southern California are unknown. We evaluated the effects of enrichment with NH4-N on mosquito production and on the nutrient uptake and growth of alkali bulrush in two studies. Overall, significantly greater numbers (> 50%) of immature mosquitoes (mainly Culex tarsalis) were found in mesocosms enriched with NH4-N than in mesocosms receiving ambient (<0.3 mg/liter) NH4-N. High NH4-N enrichment (up to 60 mg/liter) did not adversely impact the height and stem density of S. maritimus, although a significant decrease in biomass was observed at the highest enrichment level. Nitrogen uptake by alkali bulrush increased directly with NH4-N enrichment, whereas carbon was conserved in the above-ground biomass across the enrichment gradient. Alkali bulrush is recommended for use as part of integrated mosquito management programs for moderately enriched, multipurpose, constructed treatment wetlands that improve water quality as well as provide wetland habitat for waterfowl.

Tolerance to individual and joint effects of arsenic and Bacillus thuringiensis subsp. israelensis or Lysinibacillus sphaericus in Culex mosquitoes.

Arsenic contamination of global water supplies has come to the forefront in policy decisions in recent decades. However, the effects of arsenic on lower trophic levels of insects inhabiting contaminated ecosystems are not well understood. One approach to document both acute and sublethal effects of toxicants like arsenic is to assay them in combination with microbial pathogens to evaluate shifts in survival curves of the test organisms. Larvae of Culex quinquefasciatus and Culex tarsalis were reared in water containing 0 or 1 000 µg/L of arsenate or arsenite. Fourth instars were then exposed to a range of doses of Bacillus thuringiensis subsp. israelensis (Bti) or Lysinibacillus sphaericus (Ls), with shifts in lethal concentrations determined. Arsenic accumulation in 4th instars was also quantified, and a relative growth index (RGI) calculated for the treatments and compared to controls. Larvae of both species accumulated between 4 447 ± 169 ng As/g and 6 983 ± 367 ng As/g, though RGI values indicated accumulation did not affect growth and development. In all cases, the LC50 's and LC90 's of Cx. quinquefasciatus exposed jointly with arsenic and Bti/Ls were higher than Cx. tarsalis. Cx. tarsalis reared in arsenite showed a significant reduction in their Bti LC90 values compared to the control, indicating a sublethal effect of Bti. When exposed jointly with Ls, arsenite was more toxic than arsenate in Cx. tarsalis. Overall, these results indicate tolerance of these Culex species to arsenic exposures, and why this may occur is discussed.

Mtx toxins from Lysinibacillus sphaericus enhance mosquitocidal cry-toxin activity and suppress cry-resistance in Culex quinquefasciatus.

The interaction of Mtx toxins from Lysinibacillus sphaericus (formerly Bacillus sphaericus) with Bacillus thuringiensis subsp. israelensis Cry toxins and the influence of such interactions on Cry-resistance were evaluated in susceptible and Cry-resistant Culex quinquefasciatus larvae. Mtx-1 and Mtx-2 were observed to be active against both susceptible and resistant mosquitoes; however varying levels of cross-resistance toward Mtx toxins were observed in the resistant mosquitoes. A 1:1 mixture of either Mtx-1 or Mtx-2 with different Cry toxins generally showed moderate synergism, but some combinations were highly toxic to resistant larvae and suppressed resistance. Toxin synergy has been demonstrated to be a powerful tool for enhancing activity and managing Cry-resistance in mosquitoes, thus Mtx toxins may be useful as components of engineered bacterial larvicides.

Bacterial communities associated with culex mosquito larvae and two emergent aquatic plants of bioremediation importance.

Microbes are important for mosquito nutrition, growth, reproduction and control. In this study, we examined bacterial communities associated with larval mosquitoes and their habitats. Specifically, we characterized bacterial communities associated with late larval instars of the western encephalitis mosquito (Culextarsalis), the submerged portions of two emergent macrophytes (California bulrush, Schoenoplectuscalifornicus and alkali bulrush, Schoenoplectusmaritimus), and the associated water columns to investigate potential differential use of resources by mosquitoes in different wetland habitats. Using next-generation sequence data from 16S rRNA gene hypervariable regions, the alpha diversity of mosquito gut microbial communities did not differ between pond mesocosms containing distinct monotypic plants. Proteobacteria, dominated by the genus Thorsellia (Enterobacteriaceae), was the most abundant phylum recovered from C. tarsalis larvae. Approximately 49% of bacterial OTUs found in larval mosquitoes were identical to OTUs recovered from the water column and submerged portions of the two bulrushes. Plant and water samples were similar to one another, both being dominated by Actinobacteria, Bacteroidetes, Cyanobacteria, Proteobacteria and Verrucomicrobia phyla. Overall, the bacterial communities within C. tarsalis larvae were conserved and did not change across sampling dates and between two distinct plant habitats. Although Thorsellia spp. dominated mosquito gut communities, overlap of mosquito gut, plant and water-column OTUs likely reveal the effects of larval feeding. Future research will investigate the role of the key indicator groups of bacteria across the different developmental stages of this mosquito species.

Trophic Transfer of Arsenic from an Aquatic Insect to Terrestrial Insect Predators.

The movement of energy and nutrients from aquatic to terrestrial ecosystems can be substantial, and emergent aquatic insects can serve as biovectors not only for nutrients, but also for contaminants present in the aquatic environment. The terrestrial predators Tenodera aridifolia sinensis (Mantodea: Mantidae) and Tidarren haemorrhoidale (Araneae: Theridiidae) and the aquatic predator Buenoa scimitra (Hemiptera: Notonectidae) were chosen to evaluate the efficacy of arsenic transfer between aquatic and terrestrial environments. Culex tarsalis larvae were reared in either control water or water containing 1000 µg l(-1) arsenic. Adults that emerged from the control and arsenic treatments were fed to the terrestrial predators, and fourth instar larvae were fed to the aquatic predator reared in control or arsenic contaminated water. Tenodera a. sinensis fed arsenic-treated Cx. tarsalis accumulated 658±130 ng g(-1) of arsenic. There was no significant difference between control and arsenic-fed T. haemorrhoidale (range 142-290 ng g(-1)). Buenoa scimitra accumulated 5120±406 ng g(-1) of arsenic when exposed to arsenic-fed Cx. tarsalis and reared in water containing 1000 µg l(-1) arsenic. There was no significant difference between controls or arsenic-fed B. scimitra that were not exposed to water-borne arsenic, indicating that for this species environmental exposure was more important in accumulation than strictly dietary arsenic. These results indicate that transfer to terrestrial predators may play an important role in arsenic cycling, which would be particularly true during periods of mass emergence of potential insect biovectors. Trophic transfer within the aquatic environment may still occur with secondary predation, or in predators with different feeding strategies.

Micro x-ray absorption spectroscopic analysis of arsenic localization and biotransformation in Chironomus riparius Meigen (Diptera: Chironomidae) and Culex tarsalis Coquillett (Culicidae).

The distribution and speciation of arsenic (As) were analyzed in individuals of various life stages of a midge, Chironomus riparius, and the mosquito Culex tarsalis exposed to 1000 µg/l arsenate. X-ray absorption spectroscopy (XAS) revealed that C. riparius larvae accumulate As in their midgut, with inorganic arsenate [As(V)] being the predominant form, followed by arsenite [As(III)] and an As-thiol. Reduced concentrations of As in pupal and adult stages of C. riparius indicate excretion of As between the larval and pupal stages. In adults, As was limited to the thorax, and the predominant form was an As-thiol. In Cx. tarsalis, As was not found in high enough concentrations to determine As speciation, but the element was distributed throughout the larva. In adults, As was concentrated in the thorax and eyes of adults. These results have implications for understanding the biotransformation of As and its movement from aquatic to terrestrial environments.

Distribution of Culex species in vegetation bands of a constructed wetland undergoing integrated mosquito management.

The distribution and abundance of emerging Culex spp. were assessed within narrow (width: 3 m) and wide (width: 20 m) bands of California bulrush (Schoenoplectus californicus) and in the open water adjacent to emergent vegetation in 2 marshes of an ammonia-dominated wastewater treatment wetland in southern California. Emerging mosquitoes were collected along transects perpendicular to the path of water flow at 3 distances (1.5, 5, and 10 m) from the vegetation-open water interface in the wide bands of emergent vegetation, at the center of narrow bands of emergent vegetation, and at 1.5 m from the edge of emergent vegetation in the open water. The width of vegetation bands (3 vs. 20 m) influenced the effectiveness of integrated mosquito management practices, especially the application of mosquito control agents. Mosquito production from the 2 marshes also differed up to 14-fold, suggesting that the distance between the shorelines (62 vs. 74 m) of each marsh also influenced the efficacy of mosquito control agents applied from the shore and boats. Hot spots of mosquito production (75424 female Culex/m2/day) were found within the wide bands of bulrush. During summer, the relative abundance of Culex stigmatosoma among emerging mosquitoes increased from the periphery to the center of wide bands of emergent vegetation. Culex erythrothorax emergence rates were comparatively similar among the transects in the wide bands of emergent vegetation. Culex tarsalis adults increased in number from the periphery to the center of wide bands of bulrush and, in May, were > 95% of emerged mosquitoes.