Beyond competence and retention: Developing a comprehensive evaluation framework for a rural primary care nurse practitioner residency program.

ABSTRACT: With the rise in nurse practitioner (NP) residency programs, evaluations have largely focused on retention and competency completion for residents. There is a need for expanded evaluation to ensure the sustainability of NP residency programs, to ensure timely adaptations to address resident satisfaction, and to solidify a long-term pathway of NPs well prepared for rural practice. We created a family nurse practitioner (FNP) residency program with a comprehensive evaluation framework to prepare residents for practice in rural settings. The evaluation framework was developed through collaborative engagement of an external evaluation team, program leadership, and clinical site representatives. The evaluation framework of the FNP residency program combined resident assessment and holistic program evaluation, using a rapid continuous quality improvement (QI) approach. The evaluation considered three distinct perspectives: the resident, the peer coach, and the clinical site. The rapid continuous QI approach allowed program leadership to respond swiftly to programmatic challenges, improve the residency program in response to residents' reported experiences, and emphasize sustainability for continued program impact, while assessing residents' learning and performance. The program's data-driven evaluation approach has demonstrated its success in meeting the goals of the Health Resources and Services Administration funding by increasing the number of primary care providers in rural settings. The program's expansion and continued success have further validated the efficacy of this evaluation framework in assessing, improving, and ensuring the sustainability of APRN residency programs. This article calls for the adoption of similar evaluation strategies in future residency programs to promote their long-term success and impact in rural health care settings.

Relative fitness of susceptible and Cry1A.105/Cry2Ab2-single-/dual-protein-resistant Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae) on non-Bt diet and a diet containing a low concentration of two proteins.

Helicoverpa zea (Boddie) is a destructive agricultural pest species that is targeted by both Bacillus thuringiensis (Bt) maize and cotton in the United States. Cry1A.105 and Cry2Ab2 are two Bt proteins expressed in a widely planted maize event MON 89034. In this study, two tests (Test-I and Test-II) were conducted to evaluate the relative fitness of Bt-susceptible and -resistant H. zea on non-Bt diet (Test-I and Test-II) and a diet containing a mix of Cry1A.105 and Cry2Ab2 at a low concentration (Test-II only). Insect populations evaluated in Test-I were two Bt-susceptible strains and three Bt-resistant strains (a single-protein Cry1A.105-, a single-protein Cry2Ab2-, and a dual-protein Cry1A.105/Cry2Ab2-resistant strains). Test-II analyzed the same two susceptible strains, three backcrossed-and-reselected Cry1A.105/Cry2Ab2-single-/dual-protein-resistant strains, and three F1 heterozygous strains. Measurements of life table parameters showed that neither the single- nor dual-protein Cry1A.105/Cry2Ab2 resistance in H. zea was associated with fitness costs under the test conditions. The single Cry protein resistances at a concentration of a mix of Cry1A.105 and Cry2Ab2 that resulted in a zero net reproductive rate for the two susceptible strains were functionally incomplete recessive or codominant, and the dual-protein resistance was completely dominant. The lack of fitness costs could be a factor contributing to the rapid revolution of resistance to the Cry proteins in this species. Data generated from this study should aid our understanding of Cry protein resistance evolution and help in refining IRM programs for H. zea.

Characterizing the sublethal effects of SmartStax PRO dietary exposure on life history traits of the western corn rootworm, Diabrotica virgifera virgifera LeConte.

The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, is an economically important pest of field corn (Zea mays L.) across the United States (U.S.) Corn Belt. Repeated use of transgenic hybrids expressing Bacillus thuringiensis (Bt) proteins has selected for field-evolved resistance to all current rootworm-active Bt proteins. The newest product available for WCR management is SmartStax® PRO, a rootworm-active pyramid containing Cry3Bb1, Cry34/35Ab1 [now reclassified as Gpp34Ab1/Tpp35Ab1] and a new mode of action, DvSnf7 dsRNA. Understanding the fitness of adult WCR after dietary exposure to SmartStax® PRO will identify potential impacts on WCR population dynamics and inform efforts to optimize resistance management strategies. Therefore, the objective of the present study was to characterize the effect of SmartStax® PRO dietary exposure on WCR life history traits. Adult WCR were collected during 2018 and 2019 from emergence tents placed over replicated field plots of SmartStax® PRO or non-rootworm Bt corn at a site with a history of rootworm-Bt trait use and suspected resistance to Cry3Bb1 and Cry34/35Ab1. Adult survival was reduced by 97.1-99.7% in SmartStax® PRO plots relative to the non-rootworm Bt corn plots during the study. Individual male/female pairs were fed different diets of ear tissue to simulate lifetime or adult exposure. Life history parameters measured included adult longevity, adult head capsule width, lifetime female egg production, and egg viability. Results indicate that lifetime or adult exposure to SmartStax® PRO significantly reduced adult longevity and lifetime egg production. Larval exposure to SmartStax® PRO significantly reduced WCR adult size. Results from this study collectively suggest that SmartStax® PRO may negatively impact WCR life history traits, which may lead to reduced population growth when deployed in an area with WCR resistance to Bt traits.

Extended investigation of field-evolved resistance of the corn earworm Helicoverpa zea(Lepidoptera: Noctuidae) to Bacillus thuringiensis Cry1A.105 and Cry2Ab2 proteins in thesoutheastern United States.

Previous studies have reported that the corn earworm/bollworm, Helicoverpa zea (Boddie), has developed field resistance to pyramided Bacillus thuringiensis (Bt) Cry1A/Cry2A maize and cotton in certain areas of the southeastern United States. The objective of the current study was to determine the current status and distribution of the resistance to Cry1A.105 and Cry2Ab2 in H. zea. In the study, 31 H. zea populations were collected from major maize planting areas across seven southeastern states of the United States during 2018 and 2019 and assayed against the two Bt proteins. Diet over-lay bioassays showed that most of the populations collected during the two years were significantly resistant to the Cry1A.105 protein. Most of the populations collected during 2019 were also resistant to Cry2Ab2, while significant variances were observed in the susceptibility of the populations collected during 2018 to Cry2Ab2. The results showed that Cry1A.105 and Cry2Ab2 resistance in H. zea is widely distributed in the regions sampled. The resistance to Cry1A.105 appeared to have plateaued, while selection for Cry2Ab2 resistance is likely still occurring. Thus, effective measures for mitigating the Cry1A/Cry2A resistance need to be developed and implemented to ensure the sustainable use of Bt crop biotechnology.

Cry75Aa (Mpp75Aa) Insecticidal Proteins for Controlling the Western Corn Rootworm, Diabrotica virgifera virgifera, (Coleoptera: Chrysomelidae), Isolated from the Insect Pathogenic Bacteria Brevibacillus laterosporus.

This study describes three closely related proteins, cloned from Brevibacillus laterosporus strains, that are lethal upon feeding to Diabrotica virgifera virgifera LeConte, the western corn rootworm (WCR). Mpp75Aa1, Mpp75Aa2 and Mpp75Aa3 were toxic to WCR larvae when fed purified protein. Transgenic plants expressing each mMpp75Aa protein were protected from feeding damage and showed significant reduction in adult emergence from infested plants by both susceptible and Cry3Bb1 and Cry34Ab1/Cry35Ab1-resistant WCR. These results demonstrate that proteins from B. laterosporus are as efficacious as the well-known Bacillus thuringiensis (Bt) insecticidal proteins in controlling major insect pests such as WCR. The deployment of transgenic maize expressing mMpp75Aa along with other active molecules lacking cross-resistance have the potential to be a useful tool for control of WCR populations resistant to current Bt traits.IMPORTANCE Insects feeding on roots of crops can damage the plant roots resulting in yield loss due to poor water and nutrient uptake and plant lodging. In maize the western corn rootworm (WCR) can cause severe damage to the roots resulting in significant economic loss for farmers. Genetically modified (GM) expressing Bacillus thuringiensis (Bt) insect control proteins, has provided a solution for control of these pests. In recent years populations of WCR resistant to the Bt proteins in commercial GM maize have emerged. There is a need to develop new insecticidal traits for the control of WCR populations resistant to current commercial traits. New proteins with commercial level efficacy on WCR from sources other than Bt are becoming more critical. The Mpp75Aa proteins, from B. laterosporus, when expressed in maize, are efficacious against the resistant populations of WCR and have the potential to provide solutions for control of resistant WCR.

Performance of Bt-susceptible and -heterozygous dual-gene resistant genotypes of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) in seed blends of non-Bt and pyramided Bt maize.

A seed blend refuge has been implemented in the U.S. Corn Belt for Bt maize resistance management. The fall armyworm, Spodoptera frugiperda (J.E. Smith), is a target pest of Bt maize in the Americas. The larvae of this pest are mobile, which may affect the efficacy of seed blend refuges. In this study, field and greenhouse trials were conducted to determine the performance of Bt-susceptible (aabb) and -heterozygous dual-gene-resistant (AaBb) genotypes of S. frugiperda in seed blends of non-Bt and pyramided Bt maize. Three field trials evaluated larval survival, larval growth, and plant injury with aabb in seed blends of Bt maize expressing Cry1A.105/Cry2Ab2/Vip3A with 0-30% non-Bt seeds. Greenhouse tests investigated the performance of aabb and AaBb in seed blends of Cry1A.105/Cry2Ab2 with 0-30% non-Bt seeds. In pure non-Bt maize plots, after 9-13 d of neonates being released on the plants, 0.39 and 0.65 larvae/plant survived with leaf injury ratings of 4.7 and 5.9 (Davis's 1-9 scale) in the field and greenhouse, respectively. In contrast, live larvae and plant injury were virtually not observed on Bt plants across all planting patterns. Larval occurrence and plant injury by aabb on non-Bt plants were similar between seed blends and pure non-Bt plantings, suggesting that the blended refuges could provide an equivalent susceptible population as structured refuge under the test conditions. In the greenhouse, the two insect genotypes in seed blends performed similarly, indicating that the seed blends did not provide more favorable conditions for AaBb over aabb. The information generated from this study should be useful in managing S. frugiperda and evaluating if send blends could be suitable refuge options for Bt resistance management in the regions where the insect is a primary target pest.

A new Bacillus thuringiensis protein for Western corn rootworm control.

The Western corn rootworm (WCR) Diabrotica virgifera virgifera LeConte is one of the most economically important insect pests in North America. Since 2003, transgenic maize expressing WCR-active proteins from Bacillus thuringiensis (Bt) have been widely adopted as the main approach to controlling WCR in the U.S. However, the emergence of field resistance to the Bt proteins in current commercial products has been documented in recent years, highlighting the need to develop additional tools for controlling this devasting pest. Here we report the discovery of Vpb4Da2 (initially assigned as Vip4Da2), a new insecticidal protein highly selective against WCR, through high-throughput genome sequencing of a Bt strain sourced from grain dust samples collected in the eastern and central regions of the US. Vpb4Da2 contains a sequence and domain signature distinct from families of other WCR-active proteins. Under field conditions, transgenic maize expressing Vpb4Da2 demonstrates commercial-level (at or below NIS 0.25) root protection against WCR, and reduces WCR beetle emergence by ≥ 97%. Our studies also conclude that Vpb4Da2 controls WCR populations that are resistant to WCR-active transgenic maize expressing Cry3Bb1, Cry34Ab1/Cry35Ab1 (reassigned as Gpp34Ab1/Tpp35Ab1), or DvSnf7 RNA. Based on these findings, Vpb4Da2 represents a valuable new tool for protecting maize against WCR.

Physical activity impacts insulin sensitivity post metabolic bariatric surgery in adolescents with severe obesity.

BACKGROUND/OBJECTIVES: We hypothesized that physical activity (PA) improves insulin sensitivity in adolescents with severe obesity beyond that attributable to metabolic bariatric surgery (MBS). SUBJECTS/METHODS: StepWatchTM monitors objectively measured PA in 88 participants in the Teen Longitudinal Assessment of Bariatric Surgery (Teen-LABS) study. Primary outcomes included absolute change in fasting insulin, HOMA-IR, and fasting glucose from pre-surgery (baseline) to 6, 12, 24, and 36 months post-MBS. SAS PROC TRAJ generated activity trajectories based on probability and individual participant step count trajectories. Linear regression models were used, adjusted for baseline value, visit, surgical procedure, sex, and percent change in BMI. Additional models adjusted for percent change in iliac waist circumference (IWC) or percent body fat (BF), measured by bio-impedance. RESULTS: Two activity trajectories were identified: more active (MA, n = 13) and less active (LA, n = 75). MA baseline mean daily step count was >6000, increasing to >9000 at 2 years. LA mean daily step count remained at ~4000. Few participants recorded moderate step activity (cadence >80 steps/minute). Still, fasting insulin and HOMA-IR differed in association with activity trajectoy. MA was associated with a greater absolute decrease in fasting insulin (-7.8 µU/ml [95% CI: (-11.8, -3.7)], p ≤ 0.001) and a greater decrease in HOMA-IR (-1.9 [95% CI: (-3.0, -0.7)], p = 0.001), when adjusted for percent change in BMI. The significant independent effect of MA remained when adjusted for percent change in IWC or percent BF. Clinically, 100% of MA trajectory participants normalized fasting insulin, HOMA-IR, and fasting glucose by 6 months and normalization remained throughout the 36 months follow up. In contrast, 76.3 and 65.8% of LA trajectory participants normalized fasting insulin and HOMA-IR, respectively, by 12 months with 28.6% of both remaining normalized at 36 months. CONCLUSIONS: PA is independently associated with improved insulin sensitivity beyond that attributable to MBS in adolescents with severe obesity.

Inheritance of Bacillus thuringiensis Cry2Ab2 protein resistance in Helicoverpa zea (Lepidoptera: Noctuidae).

BACKGROUND: The corn earworm, Helicoverpa zea (Boddie), is a major target pest of pyramided Bt corn and cotton in the United States. Field-evolved practical resistance to Cry1 and Cry2 proteins in H. zea has been documented in multiple locations in the United States. Understanding the genetic basis of Bt resistance is essential in developing insect resistance management (IRM) strategies for the sustainable use of the Bt crop technology. In this study, we characterized the genetic bases of Cry2Ab2 resistance in H. zea using diet-overlay bioassays with two different forms of Cry2Ab2 protein. RESULTS: Laboratory bioassays using a Cry2Ab2-resistant (RR) strain, a susceptible (SS) strain, as well as cross and backcross strains, revealed that resistance to Cry2Ab2 was autosomally inherited and controlled by more than one locus. In diet bioassays, the dominance of Cry2Ab2 resistance in H. zea varied from incompletely recessive to incompletely dominant across all tested Cry2Ab2 concentrations of either Bt corn leaf powder or solubilized protein. On leaf tissue of TwinLink cotton (expressing Cry1Ab and Cry2Ae), Cry2Ab2 resistance in H. zea was completely dominant. CONCLUSION: These results have significant implications for understanding the widespread field-evolved resistance of H. zea against Cry1 and Cry2 proteins in Bt corn and cotton and should be useful in developing effective IRM strategies for H. zea. © 2020 Society of Chemical Industry.

Extended evaluation of Bt protein cross-pollination in seed blend plantings on survival, growth, and development of Helicoverpa zea feeding on refuge ears.

BACKGROUND: A seed blend, also called refuge in a bag (RIB), is used as a strategy to provide refuge for planting pyramided Bt corn in the U.S. Corn Belt. A major concern for the RIB strategy is cross-pollination between Bt and refuge plants, which may reduce susceptible populations of ear-feeding insects like Helicoverpa zea and affect the evolution of resistance. Previous studies showed a 5:95% (Non-Bt: Bt) RIB would be unlikely to provide sufficient refuge populations of H. zea for resistance management. In this study, we extended our research and conducted multiple trials to determine if three RIB plantings with greater refuge sizes (10, 20, and 30% refuge) could provide sufficient H. zea susceptible populations to delay resistance development. RESULTS: Experimental results showed that cross-pollination in 10:90%, 20:80%, and 30:70% RIB plantings still significantly reduced larval survival, delayed larval development and decreased the pupal size of H. zea on refuge plants. Regression analysis showed that the percent reduction of neonate-to-adult survivorship, relative to the survivorship on pure non-Bt ears, was significantly negatively correlated with the refuge percentage. CONCLUSION: These findings suggest that, approximately a 15% non-Bt refuge plants in RIB plantings could produce a similar number of susceptible adult moths as a 5% structured refuge planting, while an approximately 30% non-Bt refuge plant in RIB plantings could provide a similar number of susceptible moths as a 20% structured refuge planting. Information generated from this study should be useful in refining resistance management strategies for Bt crop technologies. © 2019 Society of Chemical Industry.

Physical Activity Improves Lipid and Weight-Loss Outcomes After Metabolic Bariatric Surgery in Adolescents with Severe Obesity.

OBJECTIVE: This study tested the hypothesis that physical activity improves cardiovascular disease-related lipids beyond that associated with weight loss in adolescents with severe obesity after metabolic/bariatric surgery (MBS). METHODS: Objective activity monitor data from 108 participants of the Teen Longitudinal Assessment of Bariatric Surgery (Teen-LABS) study from baseline to 3 years post MBS were used. Primary outcomes included absolute change in LDL cholesterol (LDL-C) and non-HDL cholesterol (non-HDL-C) from baseline. Baseline measurement, visit, surgical procedure, and percent change in iliac waist circumference or BMI from baseline in linear regression models were adjusted for use of generalized estimating equations. PROC TRAJ in SAS generated optimal activity trajectories based on individual step count. RESULTS: Despite low step counts and slow cadence, differences by activity trajectory were found. Greater absolute decreases in LDL-C and non-HDL-C (-15 mg/dL [95% CI: -28 to -2], P = 0.026; and -15 mg/dL [95% CI: -28 to -1], P = 0.035), respectively, were associated with more activity. More activity was associated with greater resolution of triglycerides, LDL-C, and non-HDL-C dyslipidemia and with greater weight loss 3 years post MBS. CONCLUSIONS: More activity in adolescents was associated with improvements in cardiovascular disease-related lipid measures and weight loss after MBS.

Field-evolved resistance of Helicoverpa zea (Boddie) to transgenic maize expressing pyramided Cry1A.105/Cry2Ab2 proteins in northeast Louisiana, the United States.

The corn earworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), is a major target pest of pyramided Bt maize and cotton in the U.S. In 2017 and 2018, notable ear damage and larval survival of H. zea were observed on pyramided Cry1A.105/Cry2Ab2 maize in some fields in northeast Louisiana, U.S. The objective of this study was to determine if the field control problem was due to resistance development to the Bt proteins in plants. A total of 15 H. zea populations were collected from Bt and non-Bt maize plants in 2017 and 2018 in multiple locations in Louisiana, Florida, and Georgia. Diet-overlay bioassays showed that LC50s of Cry1A.105 and Cry2Ab2 for populations collected from the areas with control problems in northeast Louisiana were as much as >1623- and 88-fold greater than that of a susceptible strain, respectively. In addition, two field trials in 2018 validated that Cry1A.105/Cry2Ab2 maize failed in managing natural H. zea populations, while Bt maize containing Vip3A was effective in northeast Louisiana. Results of the study documented that the observed field control problems of Cry1A.105/Cry2Ab2 maize against H. zea in northeast Louisiana were due to resistance development of the insect to the Bt proteins in plants. This is the first documentation of field-evolved resistance to pyramided Bt maize in a target insect species in southern U.S. However, susceptibility levels to Cry1A.105 and Cry2Ab2 varied greatly among populations collected from the three states, suggesting uneven distributions of the resistance in the region.

Phenotypic performance of nine genotypes of Cry1A.105/Cry2Ab2 dual-gene resistant fall armyworm on non-Bt and MON 89034 maize.

BACKGROUND: The fall armyworm, Spodoptera frugiperda (J.E. Smith), is the first target pest that has developed resistance to Bt crops across several countries. Leaf tissue and whole plant assays were employed to determine the survival, development, progeny production, and net reproductive rate of all nine possible genotypes of Cry1A.105/Cry2Ab2-dual gene resistant S. frugiperda on non-Bt and MON 89034 maize expressing the Cry1A.105/Cry2Ab2 proteins. RESULTS: The homozygous resistant genotype was highly resistant to Bt plants. Genotypes possessing only Cry2Ab2 resistance alleles (RAs) or two Cry1A.105 RAs only were susceptible to MON 89034 with a functional dominance level (DML ) of 0.0-0.07 on MON 89034 plants. In contrast, genotypes containing two Cry1A.105 plus one Cry2Ab2 RA performed well on Bt plants, with a DML of 0.11-0.78. Significant numbers of survivors on Bt plants were also observed for genotypes containing a single Cry1A.105 RA, or a Cry1A.105 plus one or two Cry2Ab2 RAs, with a DML of 0.0-0.47. CONCLUSIONS: The fitness of individual resistant genes on pyramided Bt plants varied in the dual-gene resistance system. Genotypes containing more Cry1A.105 RAs performed better than those possessing more Cry2Ab2 RAs. The functional dominance level of an individual resistant gene in this system is related to the DML level in its corresponding single-gene system. Data generated from this study should fill gaps in understanding dual-/multiple-gene Bt resistance, as well as providing useful information for refining resistance modeling, improving resistance risk assessment, and developing management strategies for the sustainable use of pyramided Bt maize technology. © 2019 Society of Chemical Industry.

Fitness costs of Vip3A resistance in Spodoptera frugiperda on different hosts.

BACKGROUND: Fall armyworm, Spodoptera frugiperda, is a major target pest of the Vip3A protein. The fitness of Bacillus thuringiensis (Bt)-resistant insects on refuges is an important factor in determining the speed of resistance development. Fitness costs associated with Bt resistance can vary among host plants. Here, we provide the first experimental evaluation of the interactions between non-Bt hosts and fitness costs of Vip3A resistance in S. frugiperda. RESULTS: Laboratory bioassays showed that survivorship, pupal weight, sex ratio, developmental time, fecundity, net reproductive rate, and intrinsic rate were not different among Vip3A-resistant (RR), -susceptible (SS) and -heterozygous (R♂ S♀ and R♀ S♂ ) strains of S. frugiperda on non-Bt corn, non-Bt cotton, and non-Bt diet. R♂ S♀ and R♀ S♂ also showed no differences on sorghum relative to SS. However, compared to SS, RR on sorghum showed significant reduction in pupal weight, and took longer time to develop to pupa and adult. CONCLUSION: These findings suggest that recessive fitness costs of reduced pupal weight and growth rate were detected at the individual level in the Vip3A resistant populations of S. frugiperda on sorghum. However, at the population level in terms of net reproductive rate and intrinsic rate, fitness costs of Vip3A resistance was not evident in S. frugiperda. © 2018 Society of Chemical Industry.

Performance of Bt-susceptible and -heterozygous genotypes of Spodoptera frugiperda (J.E. Smith) possessing single- or dual-gene resistance alleles in sequential feedings of non-Bt and Cry1A.105/Cry2Ab2 maize leaf tissues.

We simulated larval feeding behavior in seed blends of non-Bt and Bt maize to determine if seed blends create more favorable conditions for heterozygous-resistant insects over their Bt-susceptible counterparts. Survival, growth, development, and progeny production of four genotypes of the fall armyworm, Spodoptera frugiperda, Bt-susceptible (aabb), Cry1A.105 heterozygous resistant (Aabb), Cry2Ab2 heterozygous resistant (aaBb), and Cry1A.105/Cry2Ab2 heterozygous resistant (AaBb), were evaluated in eight feeding sequences (Seq 1-8) of non-Bt and MON89034 Bt maize leaf tissue expressing the Cry1A.105 and Cry2Ab2 proteins. We report variation in the performance of the four genotypes across the feeding sequences and biological parameters measured. Three heterozygous genotypes generally outperformed the susceptible genotype in larval survival, pupation rate, pupal weight, and progeny production. The performance was greater for Aabb over aaBb, AaBb over Aabb or aaBb, in two of the feeding sequences. The findings of this study could have important implications in assessing the risk of seed blends as refuge plantings for Bt crop resistance management where resistance in the target pest is not functionally recessive.

Fitness costs and inheritance of Cry2Ab2 resistance in Spodoptera frugiperda (J.E. Smith).

Cry2Ab2 is a Bacillus thuringiensis (Bt) protein expressed in transgenic corn and cotton targeting above-ground lepidopteran pests including the fall armyworm, Spodoptera frugiperda (J.E. Smith). The objective of this study was to characterize fitness costs and inheritance of Cry2Ab2 resistance in S. frugiperda. To determine if fitness costs were associated with the resistance, life history parameters (larval survival, growth, development and egg production) of Cry2Ab2-resistant, -susceptible, and two reciprocal F1 colonies of S. frugiperda were assayed on non-toxic diet and non-Bt corn leaf tissue. The results showed that there were no significant differences among the four insect colonies for all the biological parameters measured with few exceptions, suggesting that the resistance in the colony was not associated with significant fitness costs in the test conditions. To examine the inheritance of resistance, susceptibilities of the resistant and susceptible parents, as well as eight additional colonies generated from various genetic crosses, were assayed using Cry2Ab2-treated diet and Cry2Ab2 corn leaf tissue. The Cry2Ab2 resistance in S. frugiperda in the colony was inherited as a single autosomal recessive or incompletely recessive gene. The results of the study suggest a potential risk of resistance development in S. frugiperda to the Cry2Ab2 protein and thus effective management strategies should be implemented for the sustainable use of the Bt corn technology for pest management.

Cross-resistance to purified Bt proteins, Bt corn and Bt cotton in a Cry2Ab2-corn resistant strain of Spodoptera frugiperda.

BACKGROUND: Gene-pyramiding by combining two or more dissimilar Bacillus thuringiensis (Bt) proteins into a crop has been used to delay insect resistance. The durability of gene-pyramiding can be reduced by cross-resistance. Fall armyworm, Spodoptera frugiperda, is a major target pest of the Cry2Ab2 protein used in pyramided Bt corn and cotton. Here, we provide the first experimental evaluation of cross-resistance in S. frugiperda selected with Cry2Ab2 corn to multiple Bt sources including purified Bt proteins, Bt corn and Bt cotton. RESULTS: Concentration - response bioassays showed that resistance ratios for Cry2Ab2-resistant (RR) relative to Cry2Ab2-susceptible (SS) S. frugiperda were -1.4 for Cry1F, 1.2 for Cry1A.105, >26.7 for Cry2Ab2, >10.0 for Cry2Ae and -1.1 for Vip3A. Larvae of Cry2Ab2-heterozygous (RS), SS and RR S. frugiperda were all susceptible to Bt corn and Bt cotton containing Cry1 (Cry1F or Cry1A.105) and/or Vip3A proteins. Pyramided Bt cotton containing Cry1Ac + Cry2Ab2 or Cry1Ab + Cry2Ae were also effective against SS and RS, but not RR. CONCLUSION: These findings suggest that Cry2Ab2-corn-selected S. frugiperda is not cross-resistant to Cry1F, Cry1A.105 or Vip3A protein, or corn and cotton plants containing these Bt proteins, but it can cause strong cross-resistance to Cry2Ae and Bt crops expressing similar Bt proteins. © 2017 Society of Chemical Industry.

Cry3Bb1-Resistant Western Corn Rootworm, Diabrotica virgifera virgifera (LeConte) Does Not Exhibit Cross-Resistance to DvSnf7 dsRNA.

BACKGROUND AND METHODOLOGY: There is a continuing need to express new insect control compounds in transgenic maize against western corn rootworm, Diabrotica virgifera virgifera (LeConte) (WCR). In this study three experiments were conducted to determine cross-resistance between the new insecticidal DvSnf7 dsRNA, and Bacillus thuringiensis (Bt) Cry3Bb1; used to control WCR since 2003, with field-evolved resistance being reported. Laboratory susceptible and Cry3Bb1-resistant WCR were evaluated against DvSnf7 dsRNA in larval diet-incorporation bioassays. Additionally, the susceptibility of seven field and one field-derived WCR populations to DvSnf7 (and Cry3Bb1) was assessed in larval diet-overlay bioassays. Finally, beetle emergence of laboratory susceptible and Cry3Bb1-resistant WCR was evaluated with maize plants in the greenhouse expressing Cry3Bb1, Cry34Ab1/Cry35Ab1, or DvSnf7 dsRNA singly, or in combination. PRINCIPAL FINDINGS AND CONCLUSIONS: The Cry3Bb1-resistant colony had slight but significantly (2.7-fold; P<0.05) decreased susceptibility to DvSnf7 compared to the susceptible colony, but when repeated using a field-derived WCR population selected for reduced Cry3Bb1 susceptibility, there was no significant difference (P<0.05) in DvSnf7 susceptibility compared to that same susceptible population. Additionally, this 2.7-fold difference in susceptibility falls within the range of DvSnf7 susceptibility among the seven field populations tested. Additionally, there was no correlation between susceptibility to DvSnf7 and Cry3Bb1 for all populations evaluated. In greenhouse studies, there were no significant differences (P<0.05) between beetle emergence of susceptible and Cry3Bb1-resistant colonies on DvSnf7 and Cry34Ab1/Cry35Ab1, and between DvSnf7 and MON 87411 (DvSnf7 + Cry3Bb1) for the Cry3Bb1-resistant colony. These results demonstrate no cross-resistance between DvSnf7 and Cry3Bb1 against WCR. Therefore, pyramiding DvSnf7 with Bt proteins such as Cry3Bb1 and Cry34Ab1/Cry35Ab1 will provide a valuable IRM tool against WCR that will increase the durability of these Bt proteins. These results also illustrate the importance of using appropriate bioassay methods when characterizing field-evolved resistant WCR populations.

F2 screen for resistance to Bacillus thuringiensis Cry2Ab2-maize in field populations of Spodoptera frugiperda (Lepidoptera: Noctuidae) from the southern United States.

The fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), is a target pest of transgenic maize and cotton expressing Bacillus thuringiensis (Bt) proteins in both North and South America. In 2013 and 2014, a total of 215 F2 two-parent families of S. frugiperda were established using single-pair mating of field individuals collected from seven locations in four states of the southern U.S.: Texas, Louisiana, Georgia, and Florida. The objective of the investigation was to detect resistance alleles in field populations to Cry2Ab2, a common Bt protein produced in transgenic maize and cotton. For each F2 family, 128 F2 neonates were screened on leaf tissue of Cry2Ab2 maize plants in the laboratory. A conservative estimate of the frequency of major Cry2Ab2 resistance alleles in S. frugiperda from the four states was 0.0023 with a 95% credibility interval of 0.0003-0.0064. In addition, six families were considered to likely possess minor resistance alleles at a frequency of 0.0082 with a 95% credibility interval of 0.0033-0.0152. One F2 family from Georgia (GA-15) was confirmed to possess a major resistance allele to the Cry2Ab2 protein. Larvae from this family survived well on whole maize plants expressing Cry2Ab2 protein and demonstrated a significant level (>15-fold) of resistance when fed with the same protein incorporated in a meridic diet. The detection of the major resistance allele along with the relatively abundant minor resistance alleles revealed in this study may have important implications for resistance management.