Insect Pest Management and Environmental Risk.

Insect pests and insect pest management tactics impose risks to the environment. Environmental risk assessment is a formalized paradigm for the objective evaluation of risk in which assumptions and uncertainties are clearly presented. Therefore, a better understanding of the environmental risks and especially the comparative risks posed by insect pests and management tactics will improve integrated pest management. Risk assessments for insect pest management tactics are much more common for pesticides and genetically engineered crops than for biological control, cultural control, and semiochemicals. The reasons for this discrepancy include evidence of deleterious effects and data availability for pesticides and genetically engineered crops, public perceptions of tactics, and politics. Regardless of the regulatory oversight and frequency of risk assessments, all tactics should be subject to the risk assessment paradigm to assist in societal decisions.

Larval mosquito management and risk to aquatic ecosystems: A comparative approach including current tactics and gene-drive Anopheles techniques.

Genetic engineering of mosquitoes represents a promising tactic for reducing human suffering from malaria. Gene-drive techniques being developed that suppress or modify populations of Anopheles gambiae have the potential to be used with, or even possibly obviate, microbial and synthetic insecticides. However, these techniques are new and therefore there is attendant concern and uncertainty from regulators, policymakers, and the public about their environmental risks. Therefore, there is a need to assist decision-makers and public health stewards by assessing the risks associated with these newer mosquito management tactics so the risks can be compared as a basis for informed decision making. Previously, the effect of gene-drive mosquitoes on water quality in Africa was identified as a concern by stakeholders. Here, we use a comparative risk assessment approach for the effect of gene-drive mosquitoes on water quality in Africa. We compare the use of existing larvicides and the proposed genetic techniques in aquatic environments. Based on our analysis, we conclude that the tactic of gene-drive Anopheles for malaria management is unlikely to result in risks to aquatic environments that exceed current tactics for larval mosquitoes. As such, these new techniques would likely comply with currently recommended safety standards.

Risk Assessment for the Establishment of Vespa mandarinia (Hymenoptera: Vespidae) in the Pacific Northwest, United States.

The recent introduction of the Asian giant hornet, Vespa mandarinia Smith, in the United States in late 2019 has raised concerns about its establishment in the Pacific Northwest and its potential deleterious effects on honey bees, Apis spp., and their pollination services in the region. Therefore, we conducted a risk assessment of the establishment of V. mandarinia in Washington, Oregon, Montana, and Idaho on a county-by-county basis. Our highly conservative tier-1 qualitative and semiquantitative risk assessment relied on the biological requirements and ecological relationships of V. mandarinia in the environments of the Pacific Northwest. Our risk characterization was based on climate and habitat suitability estimates for V. mandarinia queens to overwinter and colonize nests, density and distribution of apiaries, and locations of major human-mediated introduction pathways that may increase establishment of the hornet in the counties. Our results suggest that 32 counties in the region could be at low risk, 120 at medium risk, and 23 at high risk of establishment. Many of the western counties in the region were estimated to be at the highest risk of establishment mainly because of their suitable climate for queens to overwinter, dense forest biomass for nest colonization, and proximity to major port and freight hubs in the region. By design, our tier-1 risk assessment most likely overestimates the risk of establishment, but considering its negative effects, these counties should be prioritized in ongoing monitoring and eradication efforts of V. mandarinia.

Establishing Diorhabda carinulata: Impact of Release Disturbances on Pheromone Emission and Influence of Pheromone Lures on Establishment.

Before weed biocontrol insects are transported and released in a new area, they are commonly collected into small paper containers, chilled, and kept under dark conditions. This process can be termed a pre-release protocol. The influence of a pre-release protocol on establishment success of a gregarious biological control agent was assessed using the northern tamarisk beetle, Diorhabda carinulata (Desbrochers), and its exotic, invasive host plant saltcedar (Tamarix spp.). Pre-release protocol impacts on aggregation pheromone production by D. carinulata were characterized under controlled conditions. Additional experiments were undertaken to determine if deployment of aggregation pheromone lures might enhance the agent's persistence at release sites. Adults that experienced the pre-release protocol produced less aggregation pheromone compared to undisturbed adults. Olfactometer bioassays indicated that a cohort of adults subjected to the pre-release protocol were less attractive to other adults than a control cohort. Efficacy of aggregation pheromone-based lures to retain adults at release sites was evaluated by comparing capture numbers of adult beetles at paired treatment and control release sites, 10-14 days after the release of 300, 500, or 1000 individuals. A greater number of adult D. carinulata were captured where the pheromone lures had been deployed compared to control release sites. Application of aggregation pheromone when a new release of D. carinulata is planned should allow biological control practitioners to increase retention of beetles at a release site.

Antixenosis, Antibiosis, and Potential Yield Compensatory Response in Barley Cultivars Exposed to Wheat Stem Sawfly (Hymenoptera: Cephidae) Under Field Conditions.

Wheat stem sawfly, Cephus cinctus Norton, is an economically serious pest of cereals grown in North America. Barley cultivars were previously planted as resistant crops in rotations to manage C. cinctus, but due to increasing levels of injury to this crop, this is no longer a valid management tactic in Montana. Therefore, we aimed to understand antixenosis (behavioral preference), antibiosis (mortality), and potential yield compensation (increased productivity in response to stem injuries) in barley exposed to C. cinctus. We examined these traits in eight barley cultivars. Antixenosis was assessed by counting number of eggs per stem and antibiosis was assessed by counting infested stems, dead larvae, and stems cut by mature larvae. Potential yield compensation was evaluated by comparing grain yield from three categories of stem infestation: 1) uninfested, 2) infested with dead larva, and 3) infested cut by mature larva at crop maturity. We found the greatest number of eggs per infested stem (1.80 ± 0.04), the highest proportion of infested stems (0.63 ± 0.01), and the highest proportion of cut stems (0.33 ± 0.01) in 'Hockett'. Seven out of eight cultivars had greater grain weight for infested stems than for uninfested stems. These cultivars may have compensatory responses to larval feeding injury. Overall, these barley cultivars contain varying levels of antixenosis, antibiosis, and differing levels of yield compensation. Our results provide foundational knowledge on barley traits that will provide a framework to further develop C. cinctus resistant or tolerant barley cultivars.

Host plant quantitative trait loci affect specific behavioral sequences in oviposition by a stem-mining insect.

KEY MESSAGE: Genetic diversity in quantitative loci associated with plant traits used by insects as cues for host selection can influence oviposition behavior and maternal choice. Host plant selection for oviposition is an important determinant of progeny performance and survival for phytophagous insects. Specific cues from the plant influence insect oviposition behavior; but, to date, no set of host plant quantitative trait loci (QTLs) have been shown to have an effect on behavioral sequences leading to oviposition. Three QTLs in wheat (Triticum aestivum L.) have been identified as influencing resistance to the wheat stem sawfly (WSS) (Cephus cinctus Norton). Wheat near-isogenic lines (NILs) for each of the three QTLs were used to test whether foraging WSS were able to discriminate variation in plant cues resulting from allelic changes. A QTL on chromosome 3B (Qss-msub-3BL) previously associated with stem solidness and larval antibiosis was shown to affect WSS oviposition behavior, host preference, and field infestation. Decreased preference for oviposition was also related to a QTL allele on chromosome 2D (Qwss.msub-2D). A QTL on chromosome 4A (Qwss.msub-4A.1) affected host plant attractiveness to foraging females, but did not change oviposition preference after females landed on the stem. These findings show that oviposition decisions regarding potential plant hosts require WSS females to discriminate signals from the plant associated with allelic variation at host plant quantitative loci. Allele types in a host plant QTL associated with differential survival of immature progeny can affect maternal choices for oviposition. The multidisciplinary approach used here may lead to the identification of plant genes with important community consequences, and may complement the use of antibiosis due to solid stems to control the wheat stem sawfly in agroecosystems.

The Influence of Ambient Temperature on the Susceptibility of Aedes aegypti (Diptera: Culicidae) to the Pyrethroid Insecticide Permethrin.

Insecticides are the most common strategy used for the management of mosquitoes. Changes in ambient temperature can alter the toxicity of insecticides to ectothermic organisms. Studies show organophosphate insecticides exhibit a positive correlation between ambient temperature and mortality for many insect species, and carbamate insecticides exhibit a slightly negative correlation between ambient temperature and mortality. Pyrethroid insecticides exhibit a distinctly negative correlation between increasing ambient temperature and mortality for insects. However, this relationship has not been systematically studied for adult mosquitoes. Therefore, we examined the influence of temperature on the susceptibility of adult Aedes aegypti L. (Diptera: Culicidae) when exposed to permethrin. The median lethal concentration, LC50, was estimated for adult Ae. aegypti when exposed to eight concentrations of permethrin (ranging from 0.06­0.58 ng/cm2) at each of the following temperatures­16, 23, 26, 30, 32, and 34C­for 24 h in bottle assays. The estimated LC50 for each temperature was 0.26, 0.36, 0.36, 0.45, 0.27, and 0.31 ng/cm2, respectively. Results indicated a negative correlation between temperature and mortality from 16 to 30C, a positive correlation between temperature and mortality from 30 to 32C, and a negative correlation between temperature and mortality from 32 to 34C. If mosquito populations are expanding in space and time because of increased ambient temperatures and cannot be managed as effectively with pyrethroids, the spread of mosquito-borne diseases may pose considerable additional risk to public health.

The mosquito ultra-low volume dispersion model for estimating environmental concentrations of insecticides used for adult mosquito management.

Computer models for pesticide drift are widely used tools by regulatory agencies to estimate the deposition of pesticides beyond the intended target area. Currently, there is no model in use that has been validated or verified as an accurate means of estimating concentrations of insecticides after ground-based ultra-low volume (ULV) applications used for adult mosquito management. To address the need for a validated model we created a spreadsheet-based model called Mosquito Ultra-Low Volume Dispersion (MULV-Disp) to aid in the adoption and to provide easier use of a validated model. We explain the origin, use, and utility of MULV-Disp, which can be used by regulatory agencies and other interested parties to estimate deposition of ULV insecticides.

Thermal profiles of Cicindelidia haemorrhagica (Coleoptera: Cicindelidae) activity in hot springs in Yellowstone National Park.

The wetsalts tiger beetle, Cicindelidia haemorrhagica (LeConte) (Coleoptera: Cicindelidae), is found in several active thermal hot spring areas in Yellowstone National Park (YNP) where substrate surface temperatures can exceed 50 °C. However, relationships between surface temperatures and the time adults spend on them remain poorly understood. Therefore, we characterized thermal profiles of Dragon Spring and Rabbit Creek, 2 thermally active research sites containing C. haemorrhagica in YNP, to quantify the time adults spend at different surface temperatures. We took 58 thermal video recordings of adults over 6 total days of observation ranging from 10 to 15 min for each adult. Thermal video analysis results indicated a positive relationship between the total time adult beetles spent on surface temperatures from Dragon Spring and Rabbit Creek as temperatures increased from 20 °C. Once surface temperatures exceeded 40 °C, the total time spent at those surface temperatures declined. Adults were recorded on substrates exceeding 50 °C at one of the 2 research locations. Rabbit Creek had substantially more instances of adults present with surface temperatures exceeding 40 °C, including one individual on a surface temperature of 61.5 °C. There were 3 instances of beetles spending more than 4 min at a particular surface temperature, all within the preferred range of 30-40 °C. Our thermal profile results and previous behavioral observations suggest that adults may be resistant to the heat produced from the thermal waters that influence the substrate temperatures but may not be subject to high surface temperatures as previously reported.

Heavy metal movement through insect food chains in pristine thermal springs of Yellowstone National Park.

Yellowstone National Park thermal features regularly discharge various heavy metals and metalloids. These metals are taken up by microorganisms that often form mats in thermal springs. These microbial mats also serve as food sources for invertebrate assemblages. To examine how heavy metals move through insect food webs associated with hot springs, two sites were selected for this study. Dragon-Beowulf Hot Springs, acid-sulfate chloride springs, have a pH of 2.9, water temperatures above 70 °C, and populations of thermophilic bacterial, archaeal, and algal mats. Rabbit Creek Hot Springs, alkaline springs, have a pH of up to 9, some water temperatures in excess of 60 °C, and are populated with thermophilic and phototrophic bacterial mats. Mats in both hydrothermal systems form the trophic base and support active metal transfer to terrestrial food chains. In both types of springs, invertebrates bioaccumulated heavy metals including chromium, manganese, cobalt, nickel, copper, cadmium, mercury, tin and lead, and the metalloids arsenic, selenium, and antimony resulting from consuming the algal and bacterial mat biomass. At least two orders of magnitude increase in concentrations were observed in the ephydrid shore fly Paracoenia turbida, as compared to the mats for all metals except antimony, mercury, and lead. The highest bioaccumulation factor (BAF) of 729 was observed for chromium. At the other end of the food web, the invertebrate apex predator, Cicindelidia haemorrhagica, had at least a 10-fold BAF for all metals at some location-year combinations, except with antimony. Of other taxa, high BAFs were observed with zinc for Nebria sp. (2180) and for Salda littoralis (1080). This accumulation, occurring between primary producer and primary consumer trophic levels at both springs, is biomagnified through the trophic web. These observations suggest trace metals enter the geothermal food web through the microbial mat community and are then transferred through the food chain. Also, while bioaccumulation of arsenic is uncommon, we observed five instances of increases near or exceeding 10-fold: Odontomyia sp. larvae (13.6), P. turbida (34.8), C. haemorrhagica (9.7), Rhagovelia distincta (16.3), and Ambrysus mormon (42.8).

Cowpea extrafloral nectar has potential to provide ecosystem services lost in agricultural intensification and support native parasitoids that suppress the wheat stem sawfly.

The native parasitoids Bracon cephi (Gahan) and B. lissogaster Muesebeck (Hymenoptera: Braconidae) reduce populations of Cephus cinctus Norton (Hymenoptera: Cephidae), a native grassland species, and major wheat pest on the Northern Great Plains of North America. Non-host feeding adults of these braconids increase longevity, egg load, and egg volume when provisioned carbohydrate-rich diets. Nutrition from nectar can enhance the success of natural enemies in pest management programs. Cowpea, Vigna unguiculata (L.) Walpers, is a potential cover-crop that could add resilient features to the landscape and has extrafloral nectaries (EFN), easy-access nectar sources for beneficial insects. If more cowpea was grown on the Northern Great Plains, would B. cephi and B. lissogaster benefit from foraging on putatively beneficial EFN? We investigated cowpea inflorescence stalk extrafloral nectars (IS-EFN) and leaf stipel extrafloral nectars (LS-EFN) as potential food sources for these parasitoids. Females were caged on EFN sources on living cowpea plants to assess longevity. Egg load and volume were measured at 2, 5, and 10 days after placement. Bracon cephi survived 10 days on water, 38 days on IS-EFN; B. lissogaster 6 days on water, 28 days on IS-EFN. Bracon lissogaster maintained a constant egg load and volume across treatments while B. cephi produced 2.1-fold more eggs that were 1.6-fold larger on IS-EFN. Y-tube olfactometry indicated adult females were attracted to airstreams containing cowpea volatiles. These results demonstrate that non-native, warm-season cowpea benefits these native parasitoids and may improve conservation biocontrol of C. cinctus.

Using Stable Isotopes to Determine Natal Origin and Feeding Habits of the Army Cutworm Moth, Euxoa auxiliaris (Lepidoptera: Noctuidae).

The army cutworm, Euxoa auxiliaris (Grote), is a migratory noctuid that is both an agricultural pest and an important late-season food source for grizzly bears, Ursus arctos horribilis (Linnaeus, Carnivora: Ursidae), within the Greater Yellowstone Ecosystem. Beyond the confirmation of the moths' seasonal, elevational migration in the mid-1900s, little else has been documented about their migratory patterns. To address this missing ecological component, we examined (1) migratory routes during their spring and fall migratory periods throughout their natal range, the Great Plains, and (2) natal origin at two of their summering ranges using stable hydrogen (δ2H) analyses of wings from samples collected within the areas of interest. Stable carbon (δ13C) and stable nitrogen (δ15N) analyses of wings were used to evaluate larval feeding habits of the migrants and agricultural intensity of natal origin sites, respectively. Results suggest that, rather than migrating exclusively east to west, army cutworm moths are also migrating north to south during their spring migration. Moths did not exhibit natal origin site fidelity when returning to the Great Plains. Migrants collected from the Absaroka Range had the highest probability of natal origin in Alberta, British Columbia, Saskatchewan, the most southern region of the Northwest Territories, and second highest probability of origin in Montana, Wyoming, and Idaho. Migrants collected in the Lewis Range had the highest probability of origin in the same provinces of Canada. Results suggest that migrants of the Absaroka Range fed exclusively on C3 plants as larvae and rarely fed in heavily fertilized agroecosystems.

Habitat Characteristics, Distribution, and Abundance of Cicindelidia haemorrhagica (LeConte) (Coleoptera: Cicindelidae) in Yellowstone National Park.

We observed the tiger beetle species, Cicindelidia haemorrhagica (LeConte), foraging in and reproducing near the thermal pools of Yellowstone National Park (YNP). Although this species was recorded in YNP more than 130 years ago, its distribution, ecology, and association with thermal features are unknown. Therefore, we examined the distribution and habitat characteristics of C. haemorrhagica and evaluated methods for studying its abundance. Given the extreme environments in which these beetles live, typical methods to estimate abundance are challenging. We used a series of presence/absence studies and observations to assess distribution and recorded temperature and pH measurements to determine habitat characteristics. We also conducted visual counts, light trapping, and mark/recapture experiments to assess abundance. The inability to capture C. haemorrhagica with lights led to a phototaxis experiment, which showed minimal attraction to light. Cicindelidia haemorrhagica was found throughout YNP, but it was exclusively associated with thermal springs. The thermal springs ranged from pH 2.7 to 9.0 with temperatures from 29.1 to 75.0 °C and had varying metal concentrations in soil and water. However, all thermal springs with C. haemorrhagica had barren soil with a gradual slope toward the thermal water. Specifically, habitats were thermal pools with gradual margins (a less than five-degree slope) and thermal (i.e., heated) soils for larval burrows by thermal springs or pools. Population sizes of C. haemorrhagica ranged between 500 and 1500 individuals based on visual counts.

The joint toxicity of type I, II, and nonester pyrethroid insecticides.

Evidence suggests that there are separate binding domains for type I and II pyrethroid insecticides on the voltage gated sodium channel of the nerve cell axon, but there are no studies that have examined the mixture toxicity of nonester pyrethroids and type I and II pyrethroids. Therefore, we examined the effect of nonester pyrethroid (etofenprox), type I (permethrin), and type II (cypermethrin) pyrethroid insecticides alone and in all combinations to Drosophila melanogaster Meigen. The combination of permethrin + etofenprox and permethrin + cypermethrin demonstrated antagonistic toxicity, while the combination of cypermethrin + etofenprox demonstrated synergistic toxicity. The mixture ofpermethrin + cypermethrin + etofenprox demonstrated additive toxicity. The toxicity of permethrin + cypermethrin was significantly lower than the toxicity of cypermethrin alone, but the combination was not significantly different from permethrin alone. The toxicity of permethrin + cypermethrin + etofenprox was significantly greater than the toxicity of both permethrin and etofenprox alone, but it was significantly lower than cypermethrin alone. The mixture of permethrin and etofenprox was significantly less toxic than permethrin. The explanation for the decreased toxicity observed is most likely because of the competitive binding at the voltage-gated sodium channel, which is supported by physiological and biochemical studies of pyrethroids. Our results demonstrate that the assumption that the mixture toxicity of pyrethroids would be additive is not adequate for modeling the mixture toxicity of pyrethroids to insects.

Do patterns of insect mortality in temperate and tropical zones have broader implications for insect ecology and pest management?

Background: Understanding how biotic and abiotic factors affect insect mortality is crucial for both fundamental knowledge of population ecology and for successful pest management. However, because these factors are difficult to quantify and interpret, patterns and dynamics of insect mortality remain unclear, especially comparative mortality across climate zones. Life table analysis provides robust information for quantifying population mortality and population parameters. Methods: In this study, we estimated cause-of-death probabilities and irreplaceable mortality (the portion of mortality that cannot be replaced by another cause or combination of causes) using a Multiple Decrement Life Table (MDLT) analysis of 268 insect life tables from 107 peer-reviewed journal articles. In particular, we analyzed insect mortality between temperate and tropical climate zones. Results: Surprisingly, our results suggest that non-natural enemy factors (abiotic) were the major source of insect mortality in both temperate and tropical zones. In addition, we observed that irreplaceable mortality from predators in tropical zones was 3.7-fold greater than in temperate zones. In contrast, irreplaceable mortality from parasitoids and pathogens was low and not different between temperate and tropical zones. Surprisingly, we did not observe differences in natural enemy and non-natural enemy factors based on whether the insect species was native or non-native. We suggest that characterizing predation should be a high priority in tropical conditions. Furthermore, because mortality from parasitoids was low in both tropical and temperate zones, this mortality needs to be better understood, especially as it relates to biological control and integrated pest management.

Mortality Dynamics and Life Tables of Megachile rotundata (Hymenoptera: Megachilidae), a Pollinator Managed for Alfalfa Seed Production.

The alfalfa leafcutting bee, Megachile rotundata (F.), contributes to the pollination of more than two-thirds of alfalfa used in seed production in North America. However, losses in managed populations of more than 50% are common in the United States. Thus, understanding the causes of mortality of M. rotundata is critical to find ways to maintain and increase bee populations. Over 2 yr, we identified and quantified six mortality classes of M. rotundata, as well as estimated mortality risk using the demographic life-table model, M-DEC. Research was conducted on an alfalfa field in Toston, Montana and offspring mortality was assessed in the fall of each year. Nest shelters were manipulated for a main temperature treatment (low vs high), and nest boxes inside the shelters were manipulated for a parasitism sub-treatment (parasitism-resistant vs parasitism-enabled). Total mortality was approximately 15% for both years, but the proportion for each mortality class differed substantially. Mortality increased with increasing internal nest-tunnel temperature, and nest boxes with parasitism-enabled entry had nearly double the mortality than those with parasitism-resistant entry. Based on demographic multiple decrement life table analyses, mortality from each class was highly irreplaceable. Identifying and quantifying irreplaceable mortality classes and strategies to mitigate those causes may help producers decrease total population loss of M. rotundata before the adult stage.

Plant Volatiles and Oviposition Behavior in the Selection of Barley Cultivars by Wheat Stem Sawfly (Hymenoptera: Cephidae).

Wheat stem sawfly, [Cephus cinctus (Hymenoptera: Cephidae)], females display complex behaviors for host selection and oviposition. Susceptible hollow stem wheat (Triticum aestivum L.) cultivars release a greater amount of attractive compound, (Z)-3-hexenyl acetate and receive a greater number of eggs compared to resistant solid stem wheat cultivars. However, barley (Hordeum vulgare L.) is becoming a more common host for C. cinctus in Montana. Therefore, how do host selection and oviposition behaviors on barley cultivars compare to what happens when encountering wheat cultivars? To answer this question, we carried out greenhouse experiments using two barley cultivars: 'Hockett' and 'Craft'. Between these cultivars at Zadoks stages 34 and 49, we compared host selection decisions using a Y-tube olfactometer, compared oviposition behaviors on stems, and counted the number of eggs inside individual stems. In Y-tube bioassays, we found a greater number of C. cinctus females were attracted to the airstream passing over 'Hockett' than 'Craft' barley cultivars. Although the frequencies of oviposition behaviors were similar between these cultivars, the number of eggs was greater in 'Hockett'. Volatile profiles indicated that the amount of linalool was greater in the airstream from 'Craft' than in 'Hockett' at Zadoks 34 while the amount of (Z)-3-hexenyl acetate was greater in airstream from 'Hockett' at both Zadoks 34 and 49. These results suggest that volatiles of barley plants influenced host selection behavior of ovipositing C. cinctus females, while other discriminating behaviors do not differ between cultivars.

Economic cost analysis of West Nile virus outbreak, Sacramento County, California, USA, 2005.

In 2005, an outbreak of West Nile virus (WNV) disease occurred in Sacramento County, California; 163 human cases were reported. In response to WNV surveillance indicating increased WNV activity, the Sacramento-Yolo Mosquito and Vector Control District conducted an emergency aerial spray. We determined the economic impact of the outbreak, including the vector control event and the medical cost to treat WNV disease. WNV disease in Sacramento County cost approximately $2.28 million for medical treatment and patients' productivity loss for both West Nile fever and West Nile neuroinvasive disease. Vector control cost approximately $701,790, including spray procedures and overtime hours. The total economic impact of WNV was $2.98 million. A cost-benefit analysis indicated that only 15 cases of West Nile neuroinvasive disease would need to be prevented to make the emergency spray cost-effective.

Prospective formulation of environmental risk assessments: probabilistic screening for Cry1A(b) maize risk to aquatic insects.

A critical first step for environmental risk assessment is problem formulation to identify environmental entities of concern and determinants of exposure that place these entities at risk. A conservative screening level approach was used to evaluate the potential risk to sensitive aquatic species from maize-expressed Cry1A(b) protein occurring in a representative agroecosystem. Estimated environmental concentrations for Cry1A(b) were compared to threshold concentrations of concern for putative sensitive aquatic organisms as estimated from species sensitivity distributions. The high-end risk expressed as the combined probability of short-term exposure and acute effects to a sensitive species indicated no concern in 99% of cases with limited opportunity for chronic effects due to the rapid decline of Cry1A(b) from the environment. Addressing uncertainties in the distribution of Cry1A(b) in soil, water, and sediment clarify the need for expanded ecotoxicity testing for aquatic effects.