Predicting Resistance: Quantifying the Relationship between Urban Development, Agricultural Pesticide Use, and Pesticide Resistance in a Nontarget Amphipod.

Resistance alleles within the voltage-gated sodium channel (vgsc) have been correlated with pyrethroid resistance in wild populations of the nontarget amphipod, Hyalella azteca from California (CA), U.S.A. In the present study, we expand upon the relationship between land use and the evolution of pesticide resistance in H. azteca to develop a quantitative methodology to target and screen novel populations for resistance allele genotypes in a previously uninvestigated region of the U.S. (New England: NE). By incorporating urban land development and toxicity-normalized agricultural pesticide use indices into our site selection, we successfully identified three amino acid substitutions associated with pyrethroid resistance. One of the resistance mutations has been described in H. azteca from CA (L925I). We present the remaining two (vgsc I936F and I936V) as novel pyrethroid-resistance alleles in H. azteca based on previous work in insects and elevated cyfluthrin resistance in one NE population. Our results suggest that urban pesticide use is a strong driver in the evolution of resistance alleles in H. azteca. Furthermore, our method for resistance allele screening provides an applied framework for detecting ecosystem impairment on a nationwide scale that can be incorporated into ecological risk assessment decisions.

Bioaccumulation potential of chlorpyrifos in resistant Hyalella azteca: Implications for evolutionary toxicology.

Given extensive use of pesticides in agriculture, there is concern for unintended consequences to non-target species. The non-target freshwater amphipod, Hyalella azteca has been found to show resistance to the organophosphate (OP) pesticide, chlorpyrifos, resulting from an amino acid substitution in acetylcholinesterase (AChE), suggesting a selective pressure of unintended pesticide exposure. Since resistant organisms can survive in contaminated habitats, there is potential for them to accumulate higher concentrations of insecticides, increasing the risk for trophic transfer. In the present study, we estimated the uptake and elimination of chlorpyrifos in non-resistant US Lab, and resistant Ulatis Creek (ULC Resistant), H. azteca populations by conducting 24-h uptake and 48-h elimination toxicokinetic experiments with 14C-chlorpyrifos. Our results indicated that non-resistant H. azteca had a larger uptake clearance coefficient (1467 mL g-1 h-1) than resistant animals (557 mL g-1 h-1). The half-life derived from the toxicokinetic models also estimated that steady state conditions were reached at 13.5 and 32.5 h for US Lab and ULC, respectively. Bioaccumulation was compared between non-resistant and resistant H. azteca by exposing animals to six different environmentally relevant concentrations for 28 h. Detection of chlorpyrifos in animal tissues indicated that resistant animals exposed to high concentrations of chlorpyrifos were capable of accumulating the insecticide up to 10-fold higher compared to non-resistant animals. Metabolite analysis from the 28-h concentration experiments showed that between 20 and 50 % parent compound was detected in H. azteca. These results imply that bioaccumulation potential can be more significant in chlorpyrifos resistant H. azteca and may be an essential factor in assessing the full impacts of toxicants on critical food webs, especially in the face of increasing pesticide and chemical runoff.

Pyrethroid bioaccumulation in field-collected insecticide-resistant Hyalella azteca.

Wild-type Hyalella azteca are highly sensitive to pyrethroid insecticides and typically do not survive exposure; however, pyrethroid bioaccumulation by insecticide-resistant H. azteca is an important potential risk factor for the transfer of pyrethroids to higher trophic species in aquatic systems. In the current study, four populations of pyrethroid-resistant H. azteca with corresponding sediment samples were sampled throughout the year, and nine-current use pyrethroids (tefluthrin, fenpropathrin, bifenthrin, cyhalothrin, permethrin, cyfluthrin, cypermethrin, esfenvalerate and deltamethrin) were measured. Bifenthrin was detected in every pyrethroid-resistant H. azteca tissue sample, up to 813 ng/g lipid, while cyhalothrin and permethrin were detected in fewer (18 and 28%, respectively) samples. Concurrent sampling of the sediment showed total pyrethroid concentrations exceeding toxic unit thresholds for non-resistant H. azteca survival, and confirmed the ubiquitous presence of bifenthrin at each site and sampling event. Bifenthrin concentrations in H. azteca tended to be higher in samples collected in winter months, and seasonal factors, such as temperature and rainfall, may have contributed to the noted differences in bioaccumulation. Finally, the bifenthrin and permethrin biota-sediment accumulation factors (BSAF) for pyrethroid-resistant H. azteca were similar to the BSAF values for less sensitive invertebrates, and therefore the development of resistance may enable an additional pathway for trophic transfer of pyrethroids in species that would otherwise be too sensitive to survive the exposure.

The G119S ace-1 mutation confers adaptive organophosphate resistance in a nontarget amphipod.

Organophosphate (OP) and carbamate (CM) insecticides are widely used in the United States and share the same mode of toxic action. Both classes are frequently documented in aquatic ecosystems, sometimes at levels that exceed aquatic life benchmarks. We previously identified a population of the nontarget amphipod, Hyalella azteca, thriving in an agricultural creek with high sediment levels of the OP chlorpyrifos, suggesting the population may have acquired genetic resistance to the pesticide. In the present study, we surveyed 17 populations of H. azteca in California to screen for phenotypic resistance to chlorpyrifos as well as genetic signatures of resistance in the acetylcholinesterase (ace-1) gene. We found no phenotypic chlorpyrifos resistance in populations from areas with little or no pesticide use. However, there was ~3- to 1,000-fold resistance in H. azteca populations from agricultural and/or urban areas, with resistance levels in agriculture being far higher than urban areas due to greater ongoing use of OP and CM pesticides. In every case of resistance in H. azteca, we identified a glycine-to-serine amino acid substitution (G119S) that has been shown to confer OP and CM resistance in mosquitoes and has been associated with resistance in other insects. We found that the G119S mutation was always present in a heterozygous state. Further, we provide tentative evidence of an ace-1 gene duplication in H. azteca that may play a role in chlorpyrifos resistance in some populations. The detection of a genetically based, adaptive OP and CM resistance in some of the same populations of H. azteca previously shown to harbor a genetically based adaptive pyrethroid resistance indicates that these nontarget amphipod populations have become resistant to many of the insecticides now in common use. The terrestrial application of pesticides has provided strong selective pressures to drive evolution in a nontarget, aquatic species.

Unintentional exposure to terrestrial pesticides drives widespread and predictable evolution of resistance in freshwater crustaceans.

Pesticide runoff from terrestrial environments into waterways is often lethal to freshwater organisms, but exposure may also drive evolution of pesticide resistance. We analyzed the degree of resistance and molecular genetic changes underlying resistance in Hyalella azteca, a species complex of freshwater crustaceans inadvertently exposed to pesticide pollution via runoff. We surveyed 16 waterways encompassing most major watersheds throughout California and found that land use patterns are predictive of both pyrethroid presence in aquatic sediments and pyrethroid resistance in H. azteca. Nonsynonymous amino acid substitutions in the voltage-gated sodium channel including the M918L, L925I, or L925V confer resistance in H. azteca. The most frequently identified mutation, L925I, appears to be preferred within the species complex. The L925V substitution has been associated with pyrethroid resistance in another insect, but is novel in H. azteca. We documented a variety of pyrethroid resistance mutations across several species groups within this complex, indicating that pyrethroid resistance has independently arisen in H. azteca at least six separate times. Further, the high frequency of resistance alleles indicates that pesticide-mediated selection on H. azteca populations in waterways equals or exceeds that of targeted terrestrial pests. Widespread resistance throughout California suggests current practices to mitigate off-site movement of pyrethroids are inadequate to protect aquatic life from negative ecological impacts and implies the likelihood of similar findings globally.

Are there fitness costs of adaptive pyrethroid resistance in the amphipod, Hyalella azteca?

Pyrethroid-resistant Hyalella azteca with voltage-gated sodium channel mutations have been identified at multiple locations throughout California. In December 2013, H. azteca were collected from Mosher Slough in Stockton, CA, USA, a site with reported pyrethroid (primarily bifenthrin and cyfluthrin) sediment concentrations approximately twice the 10-d LC50 for laboratory-cultured H. azteca. These H. azteca were shipped to Southern Illinois University Carbondale and have been maintained in pyrethroid-free culture since collection. Even after 22 months in culture, resistant animals had approximately 53 times higher tolerance to permethrin than non-resistant laboratory-cultured H. azteca. Resistant animals held in culture also lacked the wild-type allele at the L925 locus, and had non-synonymous substitutions that resulted in either a leucine-isoleucine or leucine-valine substitution. Additionally, animals collected from the same site nearly three years later were again resistant to the pyrethroid permethrin. When resistant animals were compared to non-resistant animals, they showed lower reproductive capacity, lower upper thermal tolerance, and the data suggested greater sensitivity to, 4, 4'-dichlorodiphenyltrichloroethane (DDT), copper (II) sulfate, and sodium chloride. Further testing of the greater heat and sodium chloride sensitivity of the resistant animals showed these effects to be unrelated to clade association. Fitness costs associated with resistance to pyrethroids are well documented in pest species (including mosquitoes, peach-potato aphids, and codling moths) and we believe that H. azteca collected from Mosher Slough also have fitness costs associated with the developed resistance.

Using Mutations for Pesticide Resistance to Identify the Cause of Toxicity in Environmental Samples.

Traditional Toxicity Identification Evaluations (TIE) are applied to identify causal agents in complex environmental samples showing toxicity and rely upon physical or chemical manipulation of samples. However, mutations conferring toxicant resistance provide the opportunity for a novel biologically based TIE. Populations within the Hyalella azteca complex from pesticide-affected waterways were 2 and 3 orders of magnitude more resistant to the pyrethroid cyfluthrin and the organophosphate chlorpyrifos, respectively, than laboratory-cultured H. azteca widely used for toxicity testing. Three resistant populations, as well as laboratory-cultured, nonresistant H. azteca, were exposed to urban and agricultural runoff. Every sample causing death or paralysis in the nonresistant individuals had no effect on pyrethroid-resistant individuals, providing strong evidence that a pyrethroid was the responsible toxicant. The lack of toxicity to chlorpyrifos-sensitive, but pyrethroid-resistant, individuals suggested chlorpyrifos was not a likely toxicant, a hypothesis supported by chemical analysis. Since these mutations that confer resistance to pesticides are highly specific, toxicity to wild-type, but not resistant animals, provides powerful evidence of causality. It may be possible to identify strains resistant to even a wider variety of toxicants, further extending the potential use of this biologically based TIE technique beyond the pyrethroid and organophosphate-resistant strains currently available.

Do pyrethroid-resistant Hyalella azteca have greater bioaccumulation potential compared to non-resistant populations? Implications for bioaccumulation in fish.

The recent discovery of pyrethroid-resistant Hyalella azteca populations in California, USA suggests there has been significant exposure of aquatic organisms to these terrestrially-applied insecticides. Since resistant organisms are able to survive in relatively contaminated habitats they may experience greater pyrethroid bioaccumulation, subsequently increasing the risk of those compounds transferring to predators. These issues were evaluated in the current study following toxicity tests in water with permethrin which showed the 96-h LC50 of resistant H. azteca (1670 ng L-1) was 53 times higher than that of non-resistant H. azteca (31.2 ng L-1). Bioaccumulation was compared between resistant and non-resistant H. azteca by exposing both populations to permethrin in water and then measuring the tissue concentrations attained. Our results indicate that resistant and non-resistant H. azteca have similar potential to bioaccumulate pyrethroids at the same exposure concentration. However, significantly greater bioaccumulation occurs in resistant H. azteca at exposure concentrations non-resistant organisms cannot survive. To assess the risk of pyrethroid trophic transfer, permethrin-dosed resistant H. azteca were fed to fathead minnows (Pimephales promelas) for four days, after which bioaccumulation of permethrin and its biotransformation products in fish tissues were measured. There were detectable concentrations of permethrin in fish tissues after they consumed dosed resistant H. azteca. These results show that bioaccumulation potential is greater in organisms with pyrethroid resistance and this increases the risk of trophic transfer when consumed by a predator. The implications of this study extend to individual fitness, populations and food webs.

Measuring and Modeling Organochlorine Pesticide Response to Activated Carbon Amendment in Tidal Sediment Mesocosms.

Activated carbon (AC) sediment amendment for hydrophobic organic contaminants (HOCs) is attracting increasing regulatory and industrial interest. However, mechanistic and well-vetted models are needed. Here, we conduct an 18 month field mesocosm trial at a site containing dichlorodiphenyltrichloroethane (DDT) and chlordane. Different AC applications were applied and, for the first time, a recently published mass transfer model was field tested under varying experimental conditions. AC treatment was effective in reducing DDT and chlordane concentration in polyethylene (PE) samplers, and contaminant extractability by Arenicola brasiliensis digestive fluids. A substantial AC particle size effect was observed. For example, chlordane concentration in PE was reduced by 93% 6 months post-treatment in the powdered AC (PAC) mesocosm, compared with 71% in the granular AC (GAC) mesocosm. Extractability of sediment-associated DDT and chlordane by A. brasiliensis digestive fluids was reduced by at least a factor of 10 in all AC treatments. The model reproduced the relative effects of varying experimental conditions (particle size, dose, mixing time) on concentrations in polyethylene passive samplers well, in most cases within 25% of experimental observations. Although uncertainties such as the effect of long-term AC fouling by organic matter remain, the study findings support the use of the model to assess long-term implications of AC amendment.

Stormwater-related transport of the insecticides bifenthrin, fipronil, imidacloprid, and chlorpyrifos into a tidal wetland, San Francisco Bay, California.

Suisun Marsh, in northern San Francisco Bay, is the largest brackish marsh in California, and provides critical habitat for many fish species. Storm runoff enters the marsh through many creeks that drain agricultural uplands and the urban areas of Fairfield and Suisun City. Five creeks were sampled throughout a major storm event in February 2014, and analyzed for representatives of several major insecticide classes. Concentrations were greatest in creeks with urban influence, though sampling was done outside of the primary season for agricultural pesticide use. Urban creek waters reached maximum concentrations of 9.9 ng/l bifenthrin, 27.4 ng/l fipronil, 11.9 ng/l fipronil sulfone, 1462 ng/l imidacloprid, and 4.0 ng/l chlorpyrifos. Water samples were tested for toxicity to Hyalella azteca and Chironomus dilutus, and while few samples caused mortality, 70% of the urban creek samples caused paralysis of either or both species. Toxic unit analysis indicated that bifenthrin was likely responsible for effects to H. azteca, and fipronil and its sulfone degradate were responsible for effects to C. dilutus. These results demonstrate the potential for co-occurrence of multiple insecticides in urban runoff, each with the potential for toxicity to particular species, and the value of toxicity monitoring using multiple species. In the channels of Suisun Marsh farther downstream, insecticide concentrations and toxicity diminished as creek waters mixed with brackish waters entering from San Francisco Bay. Only fipronil and its degradates remained measurable at 1-10 ng/l. These concentrations are not known to present a risk based on existing data, but toxicity data for estuarine and marine invertebrates, particularly for fipronil's degradates, are extremely limited.

Effects of pyrethroid insecticides in urban runoff on Chinook salmon, steelhead trout, and their invertebrate prey.

Pyrethroid insecticides can affect salmonids either indirectly through toxicity to their prey or directly by toxicity to the fish themselves. In support of a study on pyrethroid impacts to Chinook salmon and steelhead trout in the American River (Sacramento, California, USA), 96-h median effective concentration (EC50) and median lethal concentration (LC50) values for the pyrethroid bifenthrin were determined for taxa not traditionally used for toxicity testing but of interest as salmonid prey, including a chironomid, caddisflies, mayflies, and stoneflies. A laboratory was constructed on the banks of the American River to expose macroinvertebrates, Chinook salmon, and steelhead trout to flow-through river water containing urban runoff during storm events. Bifenthrin from urban runoff was found in river water following 5 rain events, reaching 14.6 ng/L. Mortality to the exposed salmonids was not observed, and sublethal effects were not seen in vitellogenin or sex steroid levels. Indirect effects via toxicity to salmonid prey are possible. Mortality to Hyalella azteca, a potential prey, was observed in every event tested, and peak bifenthrin concentrations were comparable to the 96-h EC50 of the caddisfly, Hydropsyche sp., the most important prey species on a biomass basis for American River Chinook salmon. The other invertebrates tested had EC50s exceeding bifenthrin concentrations seen in the American River, though could potentially be at risk at concentrations previously reported in smaller urban tributaries. Environ Toxicol Chem 2015;34:649-657. © 2014 SETAC.

Urban and agricultural pesticide inputs to a critical habitat for the threatened delta smelt (Hypomesus transpacificus).

The Cache Slough complex is an area of tidal sloughs in the Sacramento-San Joaquin River Delta of California (USA), and is surrounding by irrigated agricultural lands. Among the species of concern in the area is the delta smelt (Hypomesus transpacificus), a federally listed threatened species. Releases of the organophosphate insecticide chlorpyrifos and pyrethroid insecticides were examined to determine whether they represented a threat to the copepods on which delta smelt feed (Eurytemora affinis and Pseudodiaptomus forbesi) and to aquatic life in general, represented by the standard testing organism, Hyalella azteca. There was a single incident of toxicity to H. azteca as a result of discharge of agricultural irrigation water containing chlorpyrifos. Pyrethroids were not found in samples collected during the dry season. Following rain events, however, the waters of western Cache Slough repeatedly became toxic to H. azteca because of the pyrethroids bifenthrin and cyhalothrin. The 96-h median lethal concentrations (LC50s) for E. affinis and P. forbesi for the pyrethroids bifenthrin and cyhalothrin were 16.7 ng/L to 19.4 ng/L when tested at 20 °C. However, their LC50s may be 5 mg/L to 10 ng/L at in situ temperatures of the Cache Slough, comparable to the peak bifenthrin concentration observed. The dominant pyrethroid source appeared to be urban runoff entering a creek 21 km upstream of Cache Slough. Pyrethroids of urban origin were supplemented by agricultural inputs of pyrethroids and chlorpyrifos as the creek flowed toward Cache Slough.

Toxicity of the insecticide fipronil and its degradates to benthic macroinvertebrates of urban streams.

Fipronil is a phenylpyrazole insecticide with increasing urban use. Sixteen urban waterways and municipal wastewater were sampled for fipronil, its environmental degradates, and pyrethroid insecticides. Because findings could not be interpreted with existing data on fipronil degradate toxicity, EC50s and LC50s for fipronil and its sulfide and sulfone derivatives were determined for 14 macroinvertebrate species. Four species were more sensitive than any previously studied, indicating fipronil's toxicity to aquatic life has long been underestimated. The most sensitive species tested, Chironomus dilutus, had a mean 96-h EC50 of 32.5 ng/L for fipronil and 7-10 ng/L for its degradates. Hyalella azteca, a common testing species, was among the least sensitive. The typical northern California creek receiving urban stormwater runoff contains fipronil and degradate concentrations twice the EC50 of C. dilutus, and approximately one-third the EC50 for a stonefly, a caddisfly, and two mayfly species. The present study substantially increases data available on toxicity of fipronil degradates, and demonstrates that fipronil and degradates are common in urban waterways at concentrations posing a risk to a wide variety of stream invertebrates.

Predicted transport of pyrethroid insecticides from an urban landscape to surface water.

The authors developed a simple screening-level model of exposure of aquatic species to pyrethroid insecticides for the lower American River watershed (California, USA). The model incorporated both empirically derived washoff functions based on existing, small-scale precipitation simulations and empirical data on pyrethroid insecticide use and watershed properties for Sacramento County, California, USA. The authors calibrated the model to in-stream monitoring data and used it to predict daily river pyrethroid concentration from 1995 through 2010. The model predicted a marked increase in pyrethroid toxic units starting in 2000, coincident with an observed watershed-wide increase in pyrethroid use. After 2000, approximately 70% of the predicted total toxic unit exposure in the watershed was associated with the pyrethroids bifenthrin and cyfluthrin. Pyrethroid applications for aboveground structural pest control on the basis of suspension concentrate categorized product formulations accounted for greater than 97% of the predicted total toxic unit exposure. Projected application of mitigation strategies, such as curtailment of structural perimeter band and barrier treatments as recently adopted by the California Department of Pesticide Regulation, reduced predicted total toxic unit exposure by 84%. The model also predicted that similar reductions in surface-water concentrations of pyrethroids could be achieved through a switch from suspension concentrate-categorized products to emulsifiable concentrate-categorized products without restrictions on current-use practice. Even with these mitigation actions, the predicted concentration of some pyrethroids would continue to exceed chronic aquatic life criteria.

Multiple origins of pyrethroid insecticide resistance across the species complex of a nontarget aquatic crustacean, Hyalella azteca.

Use of pesticides can have substantial nonlethal impacts on nontarget species, including driving evolutionary change, often with unknown consequences for species, ecosystems, and society. Hyalella azteca, a species complex of North American freshwater amphipods, is widely used for toxicity testing of water and sediment and has frequently shown toxicity due to pyrethroid pesticides. We demonstrate that 10 populations, 3 from laboratory cultures and 7 from California water bodies, differed by at least 550-fold in sensitivity to pyrethroids. The populations sorted into four phylogenetic groups consistent with species-level divergence. By sequencing the primary pyrethroid target site, the voltage-gated sodium channel, we show that point mutations and their spread in natural populations were responsible for differences in pyrethroid sensitivity. At least one population had both mutant and WT alleles, suggesting ongoing evolution of resistance. Although nonresistant H. azteca were susceptible to the typical neurotoxic effects of pyrethroids, gene expression analysis suggests the mode of action in resistant H. azteca was not neurotoxicity but was oxidative stress sustained only at considerably higher pyrethroid concentrations. The finding that a nontarget aquatic species has acquired resistance to pesticides used only on terrestrial pests is troubling evidence of the impact of chronic pesticide transport from land-based applications into aquatic systems. Our findings have far-reaching implications for continued uncritical use of H. azteca as a principal species for monitoring and environmental policy decisions.

Pyrethroid insecticides in municipal wastewater.

Pyrethroids are widely used insecticides, but minimal information has been published on their presence in municipal wastewater in the United States. Pyrethroids in wastewater from the Sacramento, California, USA, area consisted of permethrin, bifenthrin, cypermethrin, and cyhalothrin, with a combined concentration of 200 ng/L to 500 ng/L. Sampling within the wastewater collection system leading to the treatment plant suggested pyrethroids did not originate primarily from urban runoff, but could be from any of several drain disposal practices. Wastewater from residential areas was similar in pyrethroid composition and concentration to that from the larger metropolitan area as a whole. Secondary treatment removed approximately 90% of pyrethroids, but those remaining exceeded concentrations acutely toxic to sensitive species. Toxicity to the amphipod, Hyalella azteca, was consistently evident in the final effluent. The large river into which this particular plant discharged provided sufficient dilution such that pyrethroids were undetected in the river, and there was only slight toxicity of unknown cause in 1 river sample, but effects in receiving waters elsewhere will be site-specific.

Using SPME fibers and Tenax to predict the bioavailability of pyrethroids and chlorpyrifos in field sediments.

The presence of pyrethroids in both urban and agricultural sediments at levels lethal to invertebrates has been well documented. However, variations in bioavailability among sediments make accurate predictions of toxicity based on whole sediment concentrations difficult. A proposed solution to this problem is the use of bioavailability-based estimates, such as solid phase microextraction (SPME) fibers and Tenax beads. This study compared three methods to assess the bioavailability and ultimately toxicity of pyrethroid pesticides including field-deployed SPME fibers, laboratory-exposed SPME fibers, and a 24-h Tenax extraction. The objective of the current study was to compare the ability of these methods to quantify the bioavailable fraction of pyrethroids in contaminated field sediments that were toxic to benthic invertebrates. In general, Tenax proved a more sensitive method than SPME fibers and a correlation between Tenax extractable concentrations and mortality was observed.

Identifying the cause of sediment toxicity in agricultural sediments: the role of pyrethroids and nine seldom-measured hydrophobic pesticides.

Few currently used agricultural pesticides are routinely monitored for in the environment. Even if concentrations are known, sediment LC(50) values are often lacking for common sediment toxicity testing species. To help fill this data gap, sediments in California's Central Valley were tested for nine hydrophobic pesticides seldom analyzed: abamectin, diazinon, dicofol, fenpropathrin, indoxacarb, methyl parathion, oxyfluorfen, propargite, and pyraclostrobin. Most were detected, but rarely at concentrations acutely toxic to Hyalella azteca or Chironomus dilutus. Only abamectin, fenpropathrin, and methyl parathion were found at concentrations of potential concern, and only in one or two samples. One-quarter of over 100 samples from agriculture-affected waterways exhibited toxicity, and in three-fourths of the toxic samples, pyrethroids exceeded concentrations expected to cause toxicity. The pyrethroid Bi-fen-thrin in particular, as well as lambda-cyhalothrin, cypermethrin, esfenvalerate, permethrin, and the organophosphate chlorpyrifos, were primarily responsible for the observed toxicity, rather than the more novel analytes, despite the fact that much of the sampling targeted areas of greatest use of the novel pesticides.

Pyrethroid insecticide transport into Monterey Bay through riverine suspended solids.

Pyrethroid pesticides are used widely in both agricultural and urban landscapes. Toxicity has been recorded in creeks and rivers throughout California, confirming that pyrethroids move at least short distances from the areas of terrestrial application into downstream waterways. However, their further downstream transport into the marine ecosystem has received little study. The Monterey Bay was chosen as the study system in the current project due to the close proximity of both urban centers and intense agriculture. Suspended sediments were sampled from three major rivers during storm events and showed that pyrethroids were routinely discharged from these coastal rivers, with concentrations of bifenthrin and permethrin in suspended solids of 22 and 83 ng/g, respectively. These suspended solids were deposited in estuaries and downstream reaches of rivers as they approached the coast where concentrations of pyrethroids in the sediment were greater than those expected to be toxic. However, despite their transport onto the continental shelf, pyrethroid residues were not detected in bed sediments of the shelf or in the nearby deep sea canyon, presumably due to dilution and degradation.

Stormwater input of pyrethroid insecticides to an urban river.

The American River flows for nearly 50 km through highly urbanized lands surrounding Sacramento, California, USA. Twenty-three streams, drainage canals, or pumping stations discharge urban runoff to the river, with the cumulative effect of nearly doubling the river's flow during rain events. During winter storms, the water column in the most downstream 13-km reach of the river exhibited toxicity to the standard testing species, Hyalella azteca, in 52% of samples, likely because of the pyrethroid insecticide bifenthrin. The compound is heavily used by professional pest controllers, either as a liquid perimeter treatment around homes or as granules broadcast over landscaped areas. It was found in 11 of 12 runoff sources examined, at concentrations averaging five times the H. azteca 96-h EC50. Quantified inputs of bifenthrin should have been sufficient to attain peak concentrations in the river twice those actually observed, suggesting loss by sedimentation of particulates and pesticide adsorption to the substrate and/or vegetation. Nevertheless, observed bifenthrin concentrations in the river were sufficient to cause water column toxicity, demonstrated during six storms studied over three successive winters. Toxicity and bifenthrin concentrations were greatest when river flow was low (<23 m(3) /s) but persisted even at atypically high flows (585 m(3) /s).