Management of Insect Pests with Bt Crops in the United States.

Genetically engineered corn and cotton that produce insecticidal toxins derived from the bacterium Bacillus thuringiensis (Bt) have been used to manage insect pests in the United States and elsewhere. In some cases, this has led to regional suppression of pest populations and pest eradication within the United States, and these outcomes were associated with reductions in conventional insecticides and increased profits for farmers. In other instances, pests evolved resistance to multiple Bt traits, compromising the capacity of Bt crops to manage pests and leading to increased feeding injury to crops in the field. Several aspects of pest biology and pest-crop interactions were associated with cases where pests remained susceptible versus instances where pests evolved resistance. The viability of future transgenic traits can be improved by learning from these past outcomes. In particular, efforts should be made to delay resistance by increasing the prevalence of refuges and using integrated pest management.

Susceptibility of fall armyworm, Spodoptera frugiperda (J.E.Smmith), to eight insecticides in China, with special reference to lambda-cyhalothrin.

Fall armyworm (FAW), Spodoptera frugiperda (J.E. Smith), is the main destructive insect pest of grain crops that occurs in all maize growing regions of the Americas. It has rapidly invaded the Southern China since January 2019. However, the current status of insecticide resistance in S. frugiperda has not been reported in China. In this study, we determined the susceptibility of eight populations of FAW to eight insecticides by an artificial diet incorporation method. The results showed that among eight insecticides, emamectin benzoate, spinetoram, chlorantraniliprole, chlorfenapyr, and lufenuron showed higher toxicity to this pest, while lambda-cyhalothrin and azadirachtin exhibited lower toxicity. Susceptibility of S. frugiperda to indoxacarb was significantly different (10.0-fold for LC50) across the various geographic populations. To investigate the biochemical mechanism of FAW to lambda-cyhalothrin, we performed the synergism tests and the results showed that piperonyl butoxide (PBO) and triphenyl phosphate (TPP) produced a high synergism of lambda-cyhalothrin effects in the two field populations. Sequencing of the gene encoding the acetylcholinesterase (AChE) gene in the two field populations identified two amino acid mutations, all of which have been shown previously to confer resistance to organophosphates (OPs) in several arthropod species. The results of this study provided valuable information for choosing alternative insecticides and for insecticide resistance management of S. frugiperda.

Analysis of cross-resistance to Vip3 proteins in eight insect colonies, from four insect species, selected for resistance to Bacillus thuringiensis insecticidal proteins.

Bacillus thuringiensis Vip3 proteins are synthesized and secreted during the vegetative growth phase. They are activated by gut proteases, recognize and bind to midgut receptors, form pores and lyse cells. We tested the susceptibility to Vip3Aa and Vip3Ca of Cry1A-, Cry2A-, Dipel- and Vip3-resistant insect colonies from different species to determine whether resistance to other insecticidal proteins confers cross-resistance to Vip3 proteins. As expected, the colonies resistant to Cry1A proteins, Dipel (Helicoverpa armigera, Trichoplusia ni, Ostrinia furnacalis and Plodia interpunctella) or Cry2Ab (H. armigera and T. ni) were not cross-resistant to Vip3 proteins. In contrast, H. armigera colonies resistant to Vip3Aa or Vip3Aa/Cry2Ab showed cross-resistance to the Vip3Ca protein. Moreover, the Vip3Ca protein was highly toxic to O. furnacalis (LC50 not significantly different from that of Cry1Ab), whereas the Vip3Aa protein only showed moderate growth inhibition at the highest concentration tested (100 µg/g of diet). These results extend the cross-resistance studies between Vip3 and Cry proteins, show for the first time cross-resistance between proteins within the Vip3 subfamily, and points to O. furnacalis as a target for the Vip3Ca protein.

Biological characterization and mating compatibility of Helicoverpa gelotopoeon (D.) (Lepidoptera: Noctuidae) populations from different regions in Argentina.

Helicoverpa gelotopoeon, the South American bollworm, is a polyphagous pest of the Heliothinae complex that causes damage to soybean, cotton, and chickpea crops. Some species within this complex have developed resistance to genetically modified crops and insecticides, which has led to increased interest in their genetic diversity and population structure. The objective of this study was to characterize biological and reproductive parameters of two populations of H. gelotopoeon collected in two different provinces of Argentina. Intra- and inter-population crosses revealed that H. gelotopoeon populations from both regions of Argentina did not present evidence of pre-zygotic and post-zygotic incompatibility, suggesting that Tucumán and Córdoba populations of H. gelotopoeon belong to a single wide-ranging species. Our data support the occurrence of substantial gene flow between H. gelotopoeon populations, probably due to the widely documented, long-range migratory capacity of Heliothinae species.

High frequency of CYP337B3 gene associated with control failures of Helicoverpa armigera with pyrethroid insecticides in Brazil.

Control failures with the use of pyrethroid insecticides have been reported frequently for populations of Helicoverpa armigera (Hübner) in Brazil, since its detection in 2013. Here, we confirmed and investigated the metabolic mechanisms of pyrethroid resistance in H. armigera populations from Brazil. Mortality of H. armigera populations was lower than 50% at the highest dose (10µg a.i./3rd instar larva) of the pyrethroids deltamethrin and fenvalerate in dose-response bioassays. Very low mortality (10 to 40%) was obtained at a diagnostic dose of 10µg a.i./larva for each pyrethroid in H. armigera populations collected from different agricultural regions in Brazil, from 2013 to 2016. In synergist bioassays, when larvae were treated with PBO synergist, the mortality of all populations tested was 100%. The frequency of the cytochrome P450 CYP337B3 gene was above 0.95 in all populations of H. armigera. We found only fourteen heterozygous H. armigera out of 497 individuals tested for this gene subfamily. Our results indicated that H. armigera populations from Brazil have different degrees of susceptibility to deltamethrin and fenvalerate, but all populations can be considered tolerant to pyrethroid insecticides. The chimeric P450 CYP337B3 enzyme is one of the main mechanisms of pyrethroid resistance in Brazilian H. armigera populations.

Reproductive isolation between two populations of Diatraea saccharalis (F.) (Lepidoptera: Crambidae) from different host plant species and regions in Argentina.

The sugarcane borer, Diatraea saccharalis (F.), has a widespread distribution throughout the Western Hemisphere and is a pest of many crop plants including sugarcane, corn, sorghum and rice. The use of Bacillus thuringiensis (Bt) corn has been the primary tool for managing this species in corn fields. Sugarcane borer control has been recently threatened by observations of susceptibility and/or resistance to certain varieties of Bt corn and the protein used in many newer varieties. This has led to increased interest in understanding sugarcane borer genetic diversity and gene flow within and among its populations and the consequent exchange of alleles between geographically distant populations. The objective of this study was to examine reproductive compatibility between host-associated geographic populations of D. saccharalis in Argentina and to determine whether this pest represents a complex of host-associated cryptic species rather than a wide ranging generalist species. Intra and inter-population crosses revealed that D. saccharalis populations from the northwestern and Pampas regions presented evidence of prezygotic and postzygotic incompatibility. Such a result is likely to be the product of an interruption of gene flow produced by either geographic or host plant associated isolation, suggesting that Tucumán (northwestern) and Buenos Aires (Pampas) populations of D. saccharalis are a distinct genotype and possibly an incipient species.

Early Detection and Mitigation of Resistance to Bt Maize by Western Corn Rootworm (Coleoptera: Chrysomelidae).

Transgenic Bt maize that produces less than a high-dose has been widely adopted and presents considerable insect resistance management (IRM) challenges. Western corn rootworm, Diabrotica virgifera virgifera LeConte, has rapidly evolved resistance to Bt maize in the field, leading to local loss of efficacy for some corn rootworm Bt maize events. Documenting and responding to this resistance has been complicated by a lack of rapid diagnostic bioassays and by regulatory triggers that hinder timely and effective management responses. These failures are of great concern to the scientific and agricultural community. Specific challenges posed by western corn rootworm resistance to Bt maize, and more general concerns around Bt crops that produce less than a high-dose of Bt toxin, have caused uncertainty around current IRM protocols. More than 15 years of experience with IRM has shown that high-dose and refuge-based IRM is not applicable to Bt crops that produce less than a high-dose. Adaptive IRM approaches and pro-active, integrated IRM-pest management strategies are needed and should be in place before release of new technologies that produce less than a high-dose. We suggest changes in IRM strategies to preserve the utility of corn rootworm Bt maize by 1) targeting local resistance management earlier in the sequence of responses to resistance and 2) developing area-wide criteria to address widespread economic losses. We also favor consideration of policies and programs to counteract economic forces that are contributing to rapid resistance evolution.

Binding analysis of Bacillus thuringiensis Cry1 proteins in the sugarcane borer, Diatraea saccharalis (Lepidoptera: Crambidae).

Sugarcane borer (Diatraea saccharalis, F.) is an important corn pest in South America and United States. The aim of the present study was to analyze the susceptibility and binding interactions of three Cry1A proteins and Cry1Fa in a Brazilian D. saccharalis population. The results showed that Cry1Ab was the most active, followed by Cry1Ac, Cry1Fa and Cry1Aa. All Cry1-biotinylated proteins tested bound specifically to the D. saccharalis brush border membrane vesicles (BBMV). Heterologous competition assays showed shared binding sites for all Cry1A proteins and another one shared by Cry1Fa and Cry1Ab. Thus, pyramiding Cry1Aa/Cry1Ac and Cry1F proteins would be a recommended strategy for managing this pest.

Effects of Pyramided Bt Corn and Blended Refuges on Western Corn Rootworm and Northern Corn Rootworm (Coleoptera: Chrysomelidae).

The western corn rootworm, Diabrotica virgifera virgifera LeConte, and the northern corn rootworm, Diabrotica barberi Smith & Lawrence (Coleoptera: Chrysomelidae), are major pests of corn (Zea mays L). Several transgenic corn events producing insecticidal toxins derived from the bacterium Bacillus thuringiensis (Bt) kill corn rootworm larvae and reduce injury to corn roots. However, planting of Bt corn imposes selection on rootworm populations to evolve Bt resistance. The refuge strategy and pyramiding of multiple Bt toxins can delay resistance to Bt crops. In this study, we assessed the impact of four treatments--1) non-Bt corn, 2) Cry3Bb1 corn, 3) corn pyramided with Cry3Bb1 and Cry34/35Ab1, and 4) pyramided corn with a blended refuge--on survival, time of adult emergence, and size of western and northern corn rootworm. All treatments with Bt corn led to significant reductions in the number of adults that emerged per plot. However, at one location, we identified Cry3Bb1-resistant western corn rootworm. In some cases Bt treatments reduced size of adults and delayed time of adult emergence, with effects most pronounced for pyramided corn. For both species, the number of adults that emerged from pyramided corn with a blended refuge was significantly lower than expected, based solely on emergence from pure stands of pyramided corn and non-Bt corn. The results of this study indicate that pyramided corn with a blended refuge substantially reduces survival of both western and northern corn rootworm, and as such, should be a useful tool within the context of a broader integrated pest management strategy.

Bt Maize Seed Mixtures for Helicoverpa zea (Lepidoptera: Noctuidae): Larval Movement, Development, and Survival on Non-transgenic Maize.

In 2012 and 2013, field trials were conducted near Rosemount, MN, to assess the movement and development of Helicoverpa zea (Boddie) larvae on non-Bt refuge corn plants within a seed mixture of non-Bt and Bt corn. The Bt corn hybrid expressed three Bt toxins-Cry1Ab, Cry1F, and Vip3A. As the use of seed mixtures for insect resistance management (IRM) continues to be implemented, it is necessary to further characterize how this IRM approach impacts resistance development in ear-feeding Lepidopteran pests. The potential for Bt pollen movement and cross pollination of the non-Bt ears in a seed mixture may lead to Bt toxin exposure to larvae developing on those refuge ears. Larval movement and development by H. zea, feeding on non-Bt refuge plants adjacent to either transgenic Bt or non-Bt plants, were measured to investigate the potential for unintended Bt exposure. Non-Bt plants were infested with H. zea eggs and subplots were destructively sampled twice per week within each treatment to assess larval development, location, and kernel injury. Results indicate that H. zea larval movement between plants is relatively low, ranging from 2-16% of larvae, and occurs mainly after reaching the second instar. Refuge plants in seed mixtures did not produce equivalent numbers of H. zea larvae, kernel injury, and larval development differed as compared with a pure stand of non-Bt plants. This suggests that there may be costs to larvae developing on refuge plants within seed mixtures and additional studies are warranted to define potential impacts.

Effect of Seed Blends and Soil-Insecticide on Western and Northern Corn Rootworm Emergence from mCry3A+eCry3.1Ab Bt Maize.

Seed blends containing various ratios of transgenic Bt maize (Zea mays L.) expressing the mCry3A+eCry3.1Ab proteins and non-Bt maize (near-isoline maize) were deployed alone and in combination with a soil applied pyrethroid insecticide (Force CS) to evaluate the emergence of the western corn rootworm, Diabrotica virgifera virgifera LeConte, in a total of nine field environments across the Midwestern United States in 2010 and 2011. Northern corn rootworm, Diabrotica barberi Smith & Lawrence emergence was also evaluated in four of these environments. Both western and northern corn rootworm beetle emergence from all Bt treatments was significantly reduced when compared with beetle emergence from near-isoline treatments. Averaged across all environments, western corn rootworm beetle emergence from 95:5, 90:10, and 80:20 seed blend ratios of mCry3A+eCry3.1Ab: near-isoline were 2.6-, 4.2-, and 6.7-fold greater than that from the 100:0 ratio treatment. Northern corn rootworm emergence from the same seed blend treatments resulted in 2.8-, 3.2-, and 4.2-fold more beetles than from the 100:0 treatment. The addition of Force CS (tefluthrin) significantly reduced western corn rootworm beetle emergence for each of the three treatments to which it was applied. Force CS also significantly delayed the number of days to 50% beetle emergence in western corn rootworms. Time to 50% beetle emergence in the 100% mCry3A+eCry3.1Ab treatment with Force CS was delayed 13.7 d when compared with western corn rootworm beetle emergence on near-isoline corn. These data are discussed in terms of rootworm resistance management.

Differential effects of RNAi treatments on field populations of the western corn rootworm.

RNA interference (RNAi) mediated crop protection against insect pests is a technology that is greatly anticipated by the academic and industrial pest control communities. Prior to commercialization, factors influencing the potential for evolution of insect resistance to RNAi should be evaluated. While mutations in genes encoding the RNAi machinery or the sequences targeted for interference may serve as a prominent mechanism of resistance evolution, differential effects of RNAi on target pests may also facilitate such evolution. However, to date, little is known about how variation of field insect populations could influence the effectiveness of RNAi treatments. To approach this question, we evaluated the effects of RNAi treatments on adults of three western corn rootworm (WCR; Diabrotica virgifera virgifera LeConte) populations exhibiting different levels of gut cysteine protease activity, tolerance of soybean herbivory, and immune gene expression; two populations were collected from crop rotation-resistant (RR) problem areas and one from a location where RR was not observed (wild type; WT). Our results demonstrated that RNAi targeting DvRS5 (a highly expressed cysteine protease gene) reduced gut cysteine protease activity in all three WCR populations. However, the proportion of the cysteine protease activity that was inhibited varied across populations. When WCR adults were treated with double-stranded RNA of an immune gene att1, different changes in survival among WT and RR populations on soybean diets occurred. Notably, for both genes, the sequences targeted for RNAi were the same across all populations examined. These findings indicate that the effectiveness of RNAi treatments could vary among field populations depending on their physiological and genetic backgrounds and that the consistency of an RNAi trait's effectiveness on phenotypically different populations should be considered or tested prior to wide deployment. Also, genes that are potentially subjected to differential selection in the field should be avoided for RNAi-based pest control.

Geographical and Temporal Variability in Susceptibility to Cry1F Toxin From Bacillus thuringiensis in Spodoptera frugiperda (Lepidoptera: Noctuidae) Populations in Brazil.

The genetically modified maize TC1507 event with the cry1F gene (Cry1F maize) has been used to control Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) in Brazil since the 2009-2010 cropping season. As part of the insect resistance management program, we conducted studies to determine the baseline susceptibility to Cry1F before the widespread planting of Cry1F maize. Subsequently, we evaluated the geographical and temporal variability of susceptibility to this toxin in populations of S. frugiperda collected from major maize-growing regions in Brazil. The baseline susceptibility to Cry1F was determined using a diet-overlay bioassay for a susceptible reference population and four field populations of S. frugiperda. We then monitored the susceptibility to Cry1F in 43 populations of S. frugiperda sampled in nine States of Brazil between 2011 and 2013. In the baseline study, the MIC50 (the concentration that inhibits molting to second instars in 50% of individuals) ranged from 3.59 to 72.47 ng Cry1F toxin per centimeter square. Based on the upper limit of the MIC99 value of the joint analysis from the baseline susceptibility data, the concentrations of 200 and 2,000 ng of Cry1F toxin per centimeter square were defined as diagnostic concentrations for potentially resistant individuals, and these were used to monitor the susceptibility of S. frugiperda to Cry1F. Survival at 2,000 ng Cry1F toxin per centimeter square increased significantly throughout the cropping seasons in S. frugiperda populations from São Paulo, Santa Catarina, Rio Grande do Sul, Bahia, Mato Grosso, Goiás, Mato Grosso do Sul, and Paraná. The highest survival (>50%) was reached in populations collected from Bahia, Mato Grosso, Goiás, Mato Grosso do Sul, and Paraná during the 2012-2013 cropping season. Therefore, a significant decrease in susceptibility to Cry1F was detected in S. frugiperda throughout cropping seasons, especially in regions with intensive maize production in Brazil, according to field-evolved resistance that has been demonstrated in Brazilian populations.

Is insecticide resistance a factor contributing to the increasing problems with Dalbulus maidis (Hemiptera: Cicadellidae) in Brazil?

BACKGROUND: The corn leafhopper, Dalbulus maidis, is an important pest in Brazil. While chemical control has traditionally been the cornerstone for managing this pest, field control failures have been reported for some insecticides. To understand if these failures are due to resistance, we evaluated the susceptibility of 11 field-collected populations of D. maidis to major insecticides during the 2021-2022 crop seasons in Brazil using concentration-mortality bioassays. Additionally, we employed diagnostic concentration bioassays and foliar sprays at label-recommended rates in 8-10 populations collected during the 2022-2023 crop seasons. RESULTS: High susceptibility to methomyl, carbosulfan and acephate was observed on concentration-mortality bioassays across all populations tested with resistance ratio (RR) based on LC50 <10-fold, except for one population from Bahia State that exhibited reduced susceptibility to methomyl (RR = 17.5). On the other hand, all populations exhibited reduced susceptibility to bifenthrin, acetamiprid, and imidacloprid, with RR ranging from 90 to 2000-fold. This reduced susceptibility to neonicotinoid and pyrethroid insecticides was further confirmed at diagnostic concentrations based on LC99 of the susceptible strain, with survival rates >20% and in foliar sprays with mortality rates <80%. Most populations exposed to acephate and carbosulfan exhibited low survival rates at diagnostic concentrations (<5%) and high mortality rates in foliar sprays (>80%). CONCLUSIONS: The reduced susceptibility to pyrethroid and neonicotinoid insecticides likely explain the field failures in controlling D. maidis populations in Brazil. This study represents the first large-scale susceptibility monitoring of D. maidis to insecticides, and the results will contribute to decision-making regarding the management of this pest. © 2024 Society of Chemical Industry.

Insecticide mixtures-uses, benefits and considerations.

Insecticides remain an important tool for the control of many insect pests. There has long been an interest in insecticide mixtures (in-can and tank-mix) as a means to provide the needed efficacy and/or spectrum to control many insect public health, crop pests or crop pest complexes. This aspect has become more important since insecticides developed in the last 30 years tend to be narrower in spectrum with many primarily focused on either sap-feeding or chewing insect pests. Insecticide mixtures are also seen as an important approach to insect resistance management (IRM) with certain requirements for optimal implementation. Additionally, insecticide mixtures can also address certain agronomic, commercial and intellectual property needs and opportunities. This perspective will review some of the drivers and considerations for insecticide mixtures and their potential uses. © 2024 Society of Chemical Industry.

Proactive resistance management for sustaining the efficacy of RNA interference for pest control.

Biopesticides based on RNA interference (RNAi) took a major step forward with the first registration of a sprayable RNAi product, which targets the world's most damaging potato pest. Proactive resistance management is needed to delay the evolution of resistance by pests and sustain the efficacy of RNAi biopesticides.

Larvae of Colorado potato beetle (Leptinotarsa decemlineata Say) resistant to double-stranded RNA (dsRNA) remain susceptible to small-molecule pesticides.

BACKGROUND: Implementation of resistance management tools is crucial for the continued efficacy of insect control technologies. An important aspect of insect resistance management (IRM) is the combined or sequential use of different modes-of-action to reduce selection pressure and delay evolution of resistance. This is especially important for insect pests with established ability to develop resistance to insecticides, such as the Colorado potato beetle (Leptinotarsa decemlineata, CPB). A new class of insecticides, based on double-stranded RNA (dsRNA) activating the gene silencing RNA-interference (RNAi) pathway, are currently under review for regulatory approval and commercial use in the USA against CPB. However, there is no information available on the potential for cross-resistance between RNAi insecticides and other classes of insecticides used against CPB. Herein, we aim to fill this knowledge gap by capitalizing on the availability of a CPB strain highly resistant to dsRNAs and test its susceptibility to diverse small-molecule insecticide classes compared to reference dsRNA-susceptible CPB strains. RESULTS: Differences in activity were observed among the four insecticides tested, with abamectin demonstrating highest activity against all three strains of CPB. However, no differences were observed among the dsRNA-resistant and susceptible CPB strains for any of the tested compounds. Overall, these results demonstrate lack of cross-resistance to commonly used chemical insecticides in the dsRNA-resistant strain of CPB. CONCLUSION: These data support the use of these different insecticide classes along with RNAi-based insecticides as part of an effective insect resistance management framework aimed at delaying resistance in CPB. © 2023 Society of Chemical Industry.

Larval density-dependent mortality of western corn rootworm (Coleoptera: Chrysomelidae) in Bt and non-Bt maize and implications on dose calculations†.

BACKGROUND: Transgenic crops producing insecticidal toxins from the bacterium Bacillus thuringiensis (Bt) have been used to manage insect pests for nearly 30 years. Dose of a Bt crop is key to assessing the risk of resistance evolution because it affects the heritability of resistance traits. Western corn rootworm (Diabrotica virgifera virgifera, LeConte), a major pest of maize, has evolved resistance to all commercially available Bt traits targeting it, and threatens resistance to future transgenic traits. Past research shows the dose of Bt maize targeting western corn rootworm can be confounded by larval density-dependent mortality. We conducted a 2-year field study at two locations to quantify larval density-dependent mortality in Bt and non-Bt maize. We used these results to calculate dose for our method and compared it to three previously published methods. Additionally, adult emergence and root injury were analyzed for predicting initial egg density. RESULTS: Increased pest density caused greater proportions of larvae to die in Bt maize than in non-Bt maize. All methods for calculating dose produced values less than high-dose, and stochastic variation had the greatest impact on dose at high and low pest densities. Our method for calculating dose did not produce values positively correlated with pest density while the three other methods did. CONCLUSION: To achieve the most accurate calculation of dose for transgenic maize targeting western corn rootworm, density-dependent mortality should be taken into account for both transgenic and non-transgenic maize and assessed at moderate pest densities. © 2024 Society of Chemical Industry.

Critical Facets of European Corn Borer Adult Movement Ecology Relevant to Mitigating Field Resistance to Bt-Corn.

The European corn borer (Ostrinia nubilalis, Hübner) has been managed successfully in North America since 1996 with transgenic Bt-corn. However, field-evolved resistance to all four available insecticidal Bt proteins has been detected in four provinces of Canada since 2018. Evidence suggests resistance may be spreading and evolving independently in scattered hotspots. Evolution and spread of resistance are functions of gene flow, and therefore dispersal, so design of effective resistance management and mitigation plans must take insect movement into account. Recent advances in characterizing European corn borer movement ecology have revealed a number of surprises, chief among them that a large percentage of adults disperse from the natal field via true migratory behavior, most before mating. This undermines a number of common key assumptions about adult behavior, patterns of movement, and gene flow, and stresses the need to reassess how ecological data are interpreted and how movement in models should be parameterized. While many questions remain concerning adult European corn borer movement ecology, the information currently available is coherent enough to construct a generalized framework useful for estimating the spatial scale required to implement possible Bt-resistance prevention, remediation, and mitigation strategies, and to assess their realistic chances of success.

Assessment of the 2022 post-market environmental monitoring report on the cultivation of genetically modified maize MON 810 in the EU.

Following a request from the European Commission, the European Food Safety Authority (EFSA) assessed the 2022 post-market environmental monitoring (PMEM) report on the cultivation of Cry1Ab-expressing maize event MON 810. Overall, the 2022 PMEM report provides no evidence of adverse effects of maize MON 810 cultivation. It shows a high level of compliance with refuge requirements by Spanish and Portuguese farmers growing maize MON 810, but uncertainty remains on compliance in areas where the clustered surface of maize MON 810 farms exceeds 5 ha. There are no signs of practical resistance to Cry1Ab in the field in corn borer populations collected in north-eastern Spain in 2022, although a decrease in Cry1Ab susceptibility in Mediterranean corn borer populations from this area cannot be excluded. Information retrieved through farmer questionnaires in Spain and from the scientific literature reveals no unanticipated adverse effects on human and animal health or the environment arising from the cultivation of maize MON 810. Uncertainties remain on whether 'very highly' and 'extremely' sensitive non-target lepidoptera are potentially exposed to harmful amounts of MON 810 pollen. EFSA notes that several recommendations made in the frame of the assessment of previous PMEM reports remain unaddressed and identified additional shortcomings in the 2022 PMEM report that require further consideration by the consent holder in future annual PMEM reports. Particularly, EFSA emphasises the urgent need to increase the sensitivity of the insect resistance monitoring strategy and implement mitigation measures to ensure that the exposure of non-target lepidoptera to maize MON 810 pollen is reduced to levels of no concern.