Pest population dynamics are related to a continental overwintering gradient.

Overwintering success is an important determinant of arthropod populations that must be considered as climate change continues to influence the spatiotemporal population dynamics of agricultural pests. Using a long-term monitoring database and biologically relevant overwintering zones, we modeled the annual and seasonal population dynamics of a common pest, Helicoverpa zea (Boddie), based on three overwintering suitability zones throughout North America using four decades of soil temperatures: the southern range (able to persist through winter), transitional zone (uncertain overwintering survivorship), and northern limits (unable to survive winter). Our model indicates H. zea population dynamics are hierarchically structured with continental-level effects that are partitioned into three geographic zones. Seasonal populations were initially detected in the southern range, where they experienced multiple large population peaks. All three zones experienced a final peak between late July (southern range) and mid-August to mid-September (transitional zone and northern limits). The southern range expanded by 3% since 1981 and is projected to increase by twofold by 2099 but the areas of other zones are expected to decrease in the future. These changes suggest larger populations may persist at higher latitudes in the future due to reduced low-temperature lethal events during winter. Because H. zea is a highly migratory pest, predicting when populations accumulate in one region can inform synchronous or lagged population development in other regions. We show the value of combining long-term datasets, remotely sensed data, and laboratory findings to inform forecasting of insect pests.

Regional pest suppression associated with widespread Bt maize adoption benefits vegetable growers.

Transgenic crops containing the bacterium Bacillus thuringiensis (Bt) genes reduce pests and insecticide usage, promote biocontrol services, and economically benefit growers. Area-wide Bt adoption suppresses pests regionally, with declines expanding beyond the planted Bt crops into other non-Bt crop fields. However, the offsite benefits to growers of other crops from such regional suppression remain uncertain. With data spanning 1976-2016, we demonstrate that vegetable growers benefit via decreased crop damage and insecticide applications in relation to pest suppression in the Mid-Atlantic United States. We provide evidence for the regional suppression of Ostrinia nubilalis (Hübner), European corn borer, and Helicoverpa zea (Boddie), corn earworm, populations in association with widespread Bt maize adoption (1996-2016) and decreased economic levels for injury in vegetable crops [peppers (Capsicum annuum L.), green beans (Phaseolus vulgaris L.), and sweet corn (Zea mays L., convar. saccharata)] compared with the pre-Bt period (1976-1995). Moth populations of both species significantly declined in association with widespread Bt maize (field corn) adoption, even as increased temperatures buffered the population reduction. We show marked decreases in the number of recommended insecticidal applications, insecticides applied, and O. nubilalis damage in vegetable crops in association with widespread Bt maize adoption. These offsite benefits to vegetable growers in the agricultural landscape have not been previously documented, and the positive impacts identified here expand on the reported ecological effects of Bt adoption. Our results also underscore the need to account for offsite economic benefits of pest suppression, in addition to the direct economic benefits of Bt crops.

Spatiotemporal Dynamics of the Invasive Halyomorpha halys (Hemiptera: Pentatomidae) in and Between Adjacent Corn and Soybean Fields.

Knowledge on movement and spatial patterns of insect pest populations among preferred hosts aids in the development of effective pest management strategies. In this study, we quantified the spatiotemporal dynamics of the invasive brown marmorated stink bug, Halyomorpha halys (Stål 1855), in relation to field corn, Zea mays L., and soybean, Glycine max (L.), crop phenology. We also examined the potential role of corn as a source of stink bugs in adjacent soybean. The highest density of stink bugs in each crop coincided with blister to milk-dough stages in corn (R2-R3/R4), and beginning seed to full seed (R5-R6) stages in soybean. In entire fields of adjacent corn and soybean, H. halys was found in very low density (<0.5/m(2)) or absent beyond 25 m from the field edge. Inverse distance weighted interpolations of H. halys densities suggest potential dispersal of H. halys, particularly adults and large nymphs, from corn into soybean, coinciding with the end of dough stage in corn and beginning of soybean seed development stage. These findings have important implications for managing H. halys through location and timing of scouting efforts, consideration of crop arrangement, and decisions on management interventions. Repeated scouting of field corn to assess H. halys densities, particularly from blister stage onwards, could inform decisions on management interventions for preventing or mitigating H. halys colonization into soybean. Where H. halys is an economic problem, reducing the extent of boundary shared between corn and soybean could reduce dispersal into soybean.

Effects of mowing on the arthropod community in grass buffers and adjacent crop fields.

Grass buffers are commonly planted along crop borders to filter nutrient and pesticide runoff. These buffers also provide food and shelter for beneficial and herbivorous arthropods and can serve as corridors for their movement into neighboring crops. Mowing is a common maintenance practice to control woody plants in these buffers. Field experiments were conducted to determine whether mowing influences the movement of arthropods into neighboring soybean plantings (Glycine max L) and impacts their abundance in corn (Zea mays var. indentata) the following spring. Results showed that mowing had varying effects on the abundance of herbivores, saprovores, parasitoids, and predators, particularly in the grass buffers. Aerially active arthropods in the plant canopy were more adversely affected by mowing than surface-dwelling arthropods. Mowing significantly reduced grasshopper (Orthoptera: Acrididae) density in the buffers, but did not trigger their movement into adjoining soybean fields. Parasitoids, predators, and thrips (Thysanoptera: Thripidae) were dissimilarly affected by mowing, and their responses were influenced by grass type. Altogether, these findings indicate that mowing did not cause notable movement of arthropods from grass buffers into adjoining crops and had minimal effects on the community of beneficial arthropods the following spring. Also, there were no differences in the level of insect feeding injury in crop fields next to mowed and unmowed buffers.

Field abundance and spatial patterns of ear-feeding insect pests in relation to levels of defective kernels in processing sweet corn.

In this study, we investigated the within-field distribution of sweet corn insect pests in relation to adjacent habitats and determined the level and specific causes of defective kernels affecting the quality of the final product at the processing cannery. Sap beetles [primarily Carpophilus lugubris (Murray, 1864) (Coleoptera: Nitidulidae)] and stink bugs [primarily Euschistus servus (Say) ((Heteroptera: Pentatomidae)] infested 27.6% and 73.6% of the fields, respectively. Densities of stink bugs were highest along field edges adjacent to wheat, soybean, vegetable crops, and woodlots. Levels of kernel injury were consistently higher in truckloads of ears harvested first from the outer rows. Earworm damage was confined to the ear tip and had no measurable impact on the quality of the final product. Sap beetles and blemished kernels were the major causes of defective kernels in the cannery, even though stink bugs were more abundant in the fields. Defective kernels were more positively related to physiological blemishes than to other causal factors. For all fields, defective kernel levels averaged less than 1%, resulting in excellent quality of the processed product throughout the entire season. Results provided a better understanding of the quality control issues, resulting in practical implications for improvements in field monitoring and decision-making in the cannery to minimize grading problems.

Extended Sentinel Monitoring of Helicoverpa zea Resistance to Cry and Vip3Aa Toxins in Bt Sweet Corn: Assessing Changes in Phenotypic and Allele Frequencies of Resistance.

Transgenic corn and cotton that produce Cry and Vip3Aa toxins derived from Bacillus thuringiensis (Bt) are widely planted in the United States to control lepidopteran pests. The sustainability of these Bt crops is threatened because the corn earworm/bollworm, Helicoverpa zea (Boddie), is evolving a resistance to these toxins. Using Bt sweet corn as a sentinel plant to monitor the evolution of resistance, collaborators established 146 trials in twenty-five states and five Canadian provinces during 2020-2022. The study evaluated overall changes in the phenotypic frequency of resistance (the ratio of larval densities in Bt ears relative to densities in non-Bt ears) in H. zea populations and the range of resistance allele frequencies for Cry1Ab and Vip3Aa. The results revealed a widespread resistance to Cry1Ab, Cry2Ab2, and Cry1A.105 Cry toxins, with higher numbers of larvae surviving in Bt ears than in non-Bt ears at many trial locations. Depending on assumptions about the inheritance of resistance, allele frequencies for Cry1Ab ranged from 0.465 (dominant resistance) to 0.995 (recessive resistance). Although Vip3Aa provided high control efficacy against H. zea, the results show a notable increase in ear damage and a number of surviving older larvae, particularly at southern locations. Assuming recessive resistance, the estimated resistance allele frequencies for Vip3Aa ranged from 0.115 in the Gulf states to 0.032 at more northern locations. These findings indicate that better resistance management practices are urgently needed to sustain efficacy the of corn and cotton that produce Vip3Aa.

Baseline toxicity and field efficacy of metaflumizone on Colorado potato beetle (Coleoptera: Chrysomelidae).

Baseline toxicity levels to a novel semicarbazone insecticide, metaflumizone were established for 25 field populations of Colorado potato beetle, Leptinotarsa decemlineata Say (Coleoptera: Chrysomelidae),from North America. Excluding the susceptible laboratory strain, 50% lethal concentrations of metaflumizone ranged from 0.57 to 1.31 ppm, while response slopes ranged from 1.92 to 4.24 (average = 2.93), and were unrelated to the 50% lethal concentration (r = 0.06; P = 0.76). Beetle populations with known resistance to the neonicotinoid imidacloprid also exhibited the highest LC50 levels to metaflumizone suggesting at least the possibility of cross-resistance. Additional experiments using a potato leaf-dip bioassay as well as field efficacy evaluations confirmed the high level of toxicity of metaflumizone to L. decemlineata and demonstrated a potential benefit of tank mixing a low rate of the pyrethroid esfenvalerate with metaflumizone at one-tenth the recommended field rate. These research findings confirm that metaflumizone is highly active against L. decemlineata larvae and adults and could provide an effective alternative insecticide for potato pest management.

An Evaluation of Cultural and Chemical Control Practices to Reduce Slug Damage in No-till Corn.

Slugs, primarily the gray garden slug, Deroceras reticulatum (Müller), are the most damaging non-arthropod pest of corn grown in conservation tillage systems in the US. These mollusks favor decaying plant residue on the soil surface, which provides food, shelter and optimum microenvironmental conditions for their development and survival. Here, field plot experiments evaluated several cultural and chemical control practices to suppress slug activity and feeding injury during early seedling growth. The use of row cleaners to remove surface residue over the seed row and starter fertilizer applied different ways during planting significantly reduced the percentage and severity of plants damaged by slugs by negatively affecting their activity around emerging seedlings and providing more favorable conditions for plants to outgrow and tolerate feeding injury. As rescue treatments, reduced rates of a 4% molluscicide bait applied as a directed band over the seed row, and broadcasted solutions of urea-based nitrogen applied under calm winds at night provided effective slug control. Practical considerations of these treatments are discussed, as well as changes in weather patterns and current planting practices that have had contrasting effects on slug populations and their potential damage.

Population Dynamics of Eriophyid Mites and Evaluation of Different Management Practices on Timothy Grass.

Several species of eriophyid mites are important economic pests of timothy grass in the Mid-Atlantic United States. Feeding causes stunting, curling, and brown discoloration of leaves, and yield losses ranging up to 50%. Carbaryl is the only approved chemical control for these mites. We investigated the population dynamics of field infestations, host plant resistance, and several cultural control measures to develop a more sustainable management strategy. Seasonal phenology and overall abundance differed among timothy fields and between years, with mean peak densities ranging up to 731 eggs and 1,163 mites per 2.5 cm of leaf blade. Population differences were related to the age of the field, the prevailing temperatures, and snow cover during the fall and winter months. All varieties of timothy tested were susceptible, whereas several other forage grasses were significantly resistant to eriophyid mites as possible alternatives for replacing timothy. Fall harvesting reduced the buildup of mites during the winter but populations eventually rebounded and still reached economic densities by April. Burn-down herbicide, prescribed burning, and urea-based fertilizer treatments prior to green-up in the early spring had variable effects and may help to prevent economic losses; however, several concerns about the benefit/costs and practicality of these practices are discussed.

Comparative Efficacy of Common Active Ingredients in Organic Insecticides Against Difficult to Control Insect Pests.

There exists a lack of control efficacy information to enable decision-making about which organic insecticide product works best for a given insect pest. Here, we summarize results of 153 field trials on the control efficacy of common active ingredients in organic insecticides against 12 groups of the most difficult to control insect pests. These trials evaluated primarily the organic products Entrust (spinosad), Azera (pyrethrin and azadirachtin), PyGanic (pyrethrin) and Neemix (azadirachtin), which reduced pest infestations by an overall 73.9%, 61.7%, 48.6% and 46.1% respectively, averaged across all trials. Entrust was the most effective control option for many insect pests, particularly providing >75% control of flea beetles, Colorado potato beetle, cabbageworms and alfalfa weevil, but was relatively ineffective against true bugs and aphids. Azera provided >75% control of green peach aphid, flea beetles, Japanese beetle, Mexican bean beetle, potato leafhopper and cabbageworms. PyGanic was less effective than Entrust and Azera but still provided >75% control of green peach aphid, flea beetles and potato leafhopper. The growth inhibition effects of azadirachtin in Neemix were particularly effective against larvae of Mexican bean beetle and Colorado potato beetle but was generally less effective in trials with insect infestations consisting mainly of adult stages. Those insect pests that were particularly difficult to control included thrips, stinkbugs, cucumber beetles and fruitworms. Several caveats pertaining to the application of the results are discussed.

Regional differences in gene regulation may underlie patterns of sensitivity to novel insecticides in Leptinotarsa decemlineata.

BACKGROUND: Agricultural insect pests frequently exhibit geographic variation in levels of insecticide resistance, which are often presumed to be due to the intensity of insecticide use for pest management. However, regional differences in the evolution of resistance to novel insecticides suggests that other factors are influencing rates of adaptation. We examined median lethal concentration (LC50 ) bioassay data spanning 15 years and six insecticides (abamectin, imidacloprid, spinosad, cyantraniliprole, chlorantraniliprole, and metaflumizone) for evidence of regional differences in Leptinotarsa decemlineata baseline sensitivity to insecticides as they became commercially available. RESULTS: We consistently found that larvae from Colorado potato beetle populations from the northwestern USA had the highest baseline sensitivity to novel insecticides, while populations from the eastern USA had the lowest. Comparisons of gene expression between populations from these regions revealed constitutively elevated expression of an array of detoxification genes in the East, but no evidence of additional induction when exposed to imidacloprid. CONCLUSIONS: Our results suggest a mechanism for geographic variation in rates of adaptation to insecticides, whereby baseline levels of gene expression determine a population's response to novel insecticides. These findings have implications for the regional development of insecticide resistance management strategies and for the fundamental question of what determines the rate of adaptation to insecticides. © 2020 Society of Chemical Industry.

The costs of anti-herbivore defense traits in agricultural crop plants: a case study involving leafhoppers and trichomes.

The expression of plant defenses is thought to entail costs (e.g., the allocation of resources away from growth or reproduction) that constrain the evolution of plant genotypes maximally defended against herbivores. Although central to the ecological theory underlying plant-insect interactions at large, the concept of defense costs is particularly evident in agricultural crops where plants may be under simultaneous selection for enhanced growth and/or reproduction (i.e., yield) and anti-herbivore resistance traits that deter pests. In this study we investigate the role of trichomes as a resistance mechanism against a sap-feeding insect (the leafhopper, Empoasca fabae) on potato. Natural variation in trichome density among 17 potato cultivars was used to test for the role of trichomes as a putative defense against leafhoppers, and evidence of costs in trichome expression. Two different types of costs were explored: (1) allocation costs (i.e., the relationship between trichomes and yield), and (2) costs involving trade-offs with alternative defense strategies (e.g., tolerance). Although leafhopper abundance did not decrease as trichome density increased, leafhopper injury to potato plants (foliar necrosis) was negatively correlated with trichome density. As a result, the per capita effect of leafhopper adults and nymphs on foliar damage was lower on plants with high trichome densities. We found no evidence, however, for costs of expressing this resistance trait; trichomes were not correlated with either potato yield or tolerance to herbivory. Thus, selection for multiple plant defenses to alleviate the impact of pests in agronomic crops may indeed be possible without inherent losses in plant yield.

Integrating chemical and biological control of European corn borer in bell pepper.

Using multiple locations and a series of field trials over 2 yr, we evaluated an integrated pest management program for Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) management in peppers involving biorational chemistries, inundative releases of Trichogramma ostriniae (Pang & Chen), and conservation of generalist predators. In small plot trials, three biorational insecticides (spinosad, indoxacarb, and methoxyfenozide) provided comparable control of O. nubilalis as two broad-spectrum conventional insecticides (acephate and lambda-cyhalothrin). However, lambdacyhalothrin at most locations, and indoxacarb at one location, resulted in outbreaks of green peach aphids. We also observed significant effects on the generalist predator community: beneficial communities in methoxyfenozide-treated plots were most similar to untreated controls, and acephate-treated plots were the least similar. Management systems comparing untreated controls, inundative release of T. ostriniae with methoxyfenozide applied when lepidopterans exceeded thresholds, or weekly applications of acephate or lambda-cyhalothrin, showed no effects on marketable fruit or percentage of fruit damaged, but the conventional insecticide approach caused aphid flares. Inundative releases of T. ostriniae and biorational chemistries provide a more environmentally sound approach to managing O. nubilalis in peppers, due, in part, to conservation of generalist predators.

Effects of clothianidin-treated seed on the arthropod community in a mid-Atlantic no-till corn agroecosystem.

BACKGROUND: Nearly all corn seed in the US is coated with neonicotinoid insecticides to protect against soil and foliar arthropod pests. Exposure in the soil and the systemic activity in the plant can pose non-target risks. We assessed the community-level effects of clothianidin-treated seed on the diversity and abundance of arthropod communities in a no-till corn agroecosystem over a single growing season. RESULTS: Epigeal and foliage-dwelling communities were disturbed by the clothianidin seed treatment, with significant negative and positive changes in taxa abundances. Clothianidin reduced the abundance of minute pirate bugs by 66.2%, lady beetles by 44.7%, ants by 43.4%, ground beetle adults and larvae by 31.7%, and rove beetles by 44.1% during the early corn growth stages. Herbivores, particularly thrips, were more negatively affected by clothianidin than other trophic groups. In contrast, some groups, such as collembolans and leafhoppers, exhibited significantly higher abundances in the seed treated plots. CONCLUSION: Clothianidin primarily influenced arthropod communities during the 4 weeks following planting, with disruptions to major natural enemy taxa, but communities showed trends toward recovery at the later corn stages. While the insecticide suppressed multiple herbivores, none were economically damaging to corn; thus, the pest suppression benefits of clothianidin observed in this study did not justify the non-target impacts. © 2018 Society of Chemical Industry.

Effects of insecticide-treated and Lepidopteran-active Bt transgenic sweet corn on the abundance and diversity of arthropods.

A field study was conducted over 2 yr to determine the effects of transgenic sweet corn containing a gene from the bacterium Bacillus thuringiensis (Bt) on the diversity and abundance of nontarget arthropods. The Bt hybrid (expressing Cry1Ab endotoxin for lepidopteran control) was compared with near-isogenic non-Bt and Bt hybrids treated with a foliar insecticide and with a near-isogenic non-Bt hybrid without insecticides. Plant inspections, sticky cards, and pitfall traps were used to sample a total of 573,672 arthropods, representing 128 taxonomic groups in 95 families and 18 orders. Overall biodiversity and community-level responses were not significantly affected by the transgenic hybrid. The Bt hybrid also had no significant adverse effects on population densities of specific nontarget herbivores, decomposers, and natural enemies enumerated at the family level during the crop cycle. As expected, the insecticide lambda-cyhalothrin had broad negative impacts on the abundance of many nontarget arthropods. One insecticide application in the Bt plots reduced the overall abundance of the natural enemy community by 21-48%. Five applications in the non-Bt plots reduced natural enemy communities by 33-70%. Nontarget communities affected in the insecticide-treated Bt plots exhibited some recovery, but communities exposed to five applications showed no trends toward recovery during the crop cycle. This study clearly showed that the nontarget effects of Bt transgenic sweet corn on natural enemies and other arthropods were minimal and far less than the community-level disruptions caused by lambda-cyhalothrin.

Climate change, transgenic corn adoption and field-evolved resistance in corn earworm.

Increased temperature anomaly during the twenty-first century coincides with the proliferation of transgenic crops containing the bacterium Bacillus thuringiensis (Berliner) (Bt) to express insecticidal Cry proteins. Increasing temperatures profoundly affect insect life histories and agricultural pest management. However, the implications of climate change on Bt crop-pest interactions and insect resistance to Bt crops remains unexamined. We analysed the relationship of temperature anomaly and Bt adoption with field-evolved resistance to Cry1Ab Bt sweet corn in a major pest, Helicoverpa zea (Boddie). Increased Bt adoption during 1996-2016 suppressed H. zea populations, but increased temperature anomaly buffers population reduction. Temperature anomaly and its interaction with elevated selection pressure from high Bt acreage probably accelerated the Bt-resistance development. Helicoverpa zea damage to corn ears, kernel area consumed, mean instars and proportion of late instars in Bt varieties increased with Bt adoption and temperature anomaly, through additive or interactive effects. Risk of Bt-resistant H. zea spreading is high given extensive Bt adoption, and the expected increase in overwintering and migration. Our study highlights the challenges posed by climate change for Bt biotechnology-based agricultural pest management, and the need to incorporate evolutionary processes affected by climate change into Bt-resistance management programmes.

Correction: Field-Evolved Resistance in Corn Earworm to Cry Proteins Expressed by Transgenic Sweet Corn.

[This corrects the article DOI: 10.1371/journal.pone.0169115.].

Correction: Assessment of Chronic Sublethal Effects of Imidacloprid on Honey Bee Colony Health.

[This corrects the article DOI: 10.1371/journal.pone.0118748.].

Host-plant-mediated competition via induced resistance: interactions between pest herbivores on potatoes.

Plant-mediated competition among insect herbivores occurs when one species induces changes in plant chemistry, nutrition, or morphology that render plants resistant to attack by others. We explored plant-mediated interspecific interactions between the potato leafhopper (Empoasca fabae) and the Colorado potato beetle (Leptinotarsa decemlineata), two important pests on potatoes. Leafhoppers colonize fields in advance of beetles, and thus the possibility exists that previous feeding by leafhoppers induces changes in potato plants that have adverse consequences for beetles. The consequences of leafhopper-induced resistance for beetle performance were studied in the greenhouse, field cages, and in large open-field plots. Potato plants were exposed to four densities of leafhoppers (none, low, moderate, and high), and visible feeding symptoms were measured as percentage leaf curling, chlorosis, and necrosis. The oviposition preference, performance, and survivorship of Colorado potato beetles were then measured on the four categories of induced plants in field-cage and greenhouse settings. In open field plots, survival on the four categories of induced plants was determined by placing cohorts of beetle adults onto plants and measuring the densities of resulting eggs, larvae, and emerging Fl adults. Leafhopper-induced symptoms on potato plants were density dependent, with the percentage of curled, chlorotic, and necrotic leaves increasing with leafhopper density. Previous feeding by leafhoppers adversely affected oviposition and larval performance of beetles. Fewer egg masses were deposited on plants that incurred high levels of leafhopper feeding. Similarly, larval development was delayed and emerging adult beetles weighed less when fed induced foliage from the high leafhopper-density treatment. Beetles survived less well in the field on plants experiencing moderate and high levels of leafhopper feeding as evidenced by lower densities of eggs, larvae, and emerging F1 adults. Overall, leafhoppers and beetles competed through feeding-induced changes in plant quality. Notably, the asymmetric interaction took place at a large spatial scale in open field plots and had negative consequences that persisted to the next beetle generation. Ultimately, to establish an effective management strategy for crop pests such as leafhoppers, it is essential to consider the positive indirect effects of induced resistance along with the negative direct effects on crop yield.

Field-Evolved Resistance in Corn Earworm to Cry Proteins Expressed by Transgenic Sweet Corn.

BACKGROUND: Transgenic corn engineered with genes expressing insecticidal toxins from the bacterium Bacillus thuringiensis (Berliner) (Bt) are now a major tool in insect pest management. With its widespread use, insect resistance is a major threat to the sustainability of the Bt transgenic technology. For all Bt corn expressing Cry toxins, the high dose requirement for resistance management is not achieved for corn earworm, Helicoverpa zea (Boddie), which is more tolerant to the Bt toxins. METHODOLOGY/MAJOR FINDINGS: We present field monitoring data using Cry1Ab (1996-2016) and Cry1A.105+Cry2Ab2 (2010-2016) expressing sweet corn hybrids as in-field screens to measure changes in field efficacy and Cry toxin susceptibility to H. zea. Larvae successfully damaged an increasing proportion of ears, consumed more kernel area, and reached later developmental stages (4th - 6th instars) in both types of Bt hybrids (Cry1Ab-event Bt11, and Cry1A.105+Cry2Ab2-event MON89034) since their commercial introduction. Yearly patterns of H. zea population abundance were unrelated to reductions in control efficacy. There was no evidence of field efficacy or tissue toxicity differences among different Cry1Ab hybrids that could contribute to the decline in control efficacy. Supportive data from laboratory bioassays demonstrate significant differences in weight gain and fitness characteristics between the Maryland H. zea strain and a susceptible strain. In bioassays with Cry1Ab expressing green leaf tissue, Maryland H. zea strain gained more weight than the susceptible strain at all concentrations tested. Fitness of the Maryland H. zea strain was significantly lower than that of the susceptible strain as indicated by lower hatch rate, longer time to adult eclosion, lower pupal weight, and reduced survival to adulthood. CONCLUSIONS/SIGNIFICANCE: After ruling out possible contributing factors, the rapid change in field efficacy in recent years and decreased susceptibility of H. zea to Bt sweet corn provide strong evidence of field-evolved resistance in H. zea populations to multiple Cry toxins. The high adoption rate of Bt field corn and cotton, along with the moderate dose expression of Cry1Ab and related Cry toxins in these crops, and decreasing refuge compliance probably contributed to the evolution of resistance. Our results have important implications for resistance monitoring, refuge requirements and other regulatory policies, cross-resistance issues, and the sustainability of the pyramided Bt technology.