Significance Crops genetically engineered to produce insecticidal toxins derived from the bacterium Bacillus thuringiensis (Bt) kill pest insects and reduce the use of conventional insecticides. However, the evolution of Bt resistance can diminishes these benefits. The western corn rootworm is a serious pest of maize and is managed with Bt maize. Beginning in 2009, western corn rootworm with resistance to maize producing the Bt toxin Cry3Bb1 imposed severe injury to Cry3Bb1 maize in Iowa. We show that cross-resistance exists between Cry3Bb1 maize and mCry3A maize and is associated with severe injury to Bt maize in farmers’ fields. These results illustrate that Bt crops producing less than a high dose of toxin against target pests may select for resistance rapidly; consequently, current approaches for managing Bt resistance should be reexamined.

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Western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), and northern corn rootworm, Diabrotica barberi Smith & Lawrence, are major pests of corn (Zea mays L.). Corn producing Bacillus thuringiensis (Bt) toxins are widely used to manage Diabrotica spp.; however, Bt resistance by D. v. virgifera has led to high levels of feeding injury in the field. We tested whether field history affected root injury and abundance of adult Diabrotica spp. In 2013 and 2014, four types of cornfields were sampled: 1) recently rotated fields, 2) continuous cornfields, 3) fields with a history of injury to Bt corn (past problem fields), and 4) fields with greater than one node of injury to Bt corn at the time of sampling (current problem fields). Data were collected on field history, root injury, and the abundance of adult Diabrotica spp. from each field. Root injury and the abundance of D. v. virgifera were significantly greater in current problem fields compared to the other field types, while D. barberi were significantly more abundant in recently rotated fields. Root injury and the abundance of D. v. virgifera did not differ among recently rotated fields, continuous cornfields, and past problem fields. Analysis of field history showed that recently rotated fields were characterized by significantly less Bt corn, soil-applied insecticides, and years planted to corn continuously. These results suggest that greater cropping practice diversity can reduce management inputs for Diabrotica spp.; however, its effects on resistance evolution remain undetermined.

Challenges in maintaining high effectiveness of classic vector control in urban areas has renewed the interest in indoor residual spraying (IRS) as a promising approach for Aedes -borne disease prevention. While IRS has many benefits, application time and intrusive indoor applications make its scalability in urban areas difficult. Modifying IRS to account for Ae . aegypti resting behavior, named targeted IRS (TIRS, spraying walls below 1.5 m and under furniture) can reduce application time; however, an untested assumption is that modifications to IRS will not negatively impact entomological efficacy. We conducted a comparative experimental study evaluating the residual efficacy of classically-applied IRS (as developed for malaria control) compared to two TIRS application methods using a carbamate insecticide against a pyrethroid-resistant, field-derived Ae . aegypti strain. We performed our study within a novel experimental house setting (n = 9 houses) located in Merida (Mexico), with similar layouts and standardized contents. Classic IRS application (insecticide applied to full walls and under furniture) was compared to: a) TIRS: insecticide applied to walls below 1.5 m and under furniture, and b) Resting Site TIRS (RS-TIRS): insecticide applied only under furniture. Mosquito mortality was measured eight times post-application (out to six months post-application) by releasing 100 Ae . aegypti females /house and collecting live and dead individuals after 24 hrs exposure. Compared to Classic IRS, TIRS and RS-TIRS took less time to apply (31% and 82% reduction, respectively) and used less insecticide (38% and 85% reduction, respectively). Mortality of pyrethroid-resistant Ae . aegypti did not significantly differ among the three IRS application methods up to two months post application, and did not significantly differ between Classic IRS and TIRS up to four months post application. These data illustrate that optimizing IRS to more efficiently target Ae . aegypti can both reduce application time and insecticide volume with no apparent reduction in entomological efficacy.

Western corn rootworm, Diabrotica virgifera virgifera LeConte, has evolved resistance to transgenic maize, Zea maize L., that produces the insecticidal protein Cry3Bb1, which is derived from the bacterium Bacillus thuringiensis. We hypothesized that the level of Cry3Bb1 resistance in populations of western corn rootworm could be influenced by farming practices. To test this hypothesis, we evaluated the effect of field history on resistance to Cry3Bb1 maize by western corn rootworm. In 2013 and 2014, rootworm adults were collected from the four types of maize fields: 1) current problem fields, 2) past problem fields, 3) rotated maize fields, and 4) continuous maize fields. Those field populations along with seven Bt-susceptible control populations were tested for Cry3Bb1 resistance with both plant-based and diet-based bioassays. All field populations were resistant to Cry3Bb1 regardless of field history, however, some variation in the degree of resistance was found. For all categories of field populations, larval survivorship on Cry3Bb1 maize was significantly higher than control populations, and did not differ from survival on non-Bt maize. Evidence of resistance to Cry3Bb1 maize in plant-based bioassays was further supported by diet-based bioassays and we found a positive relationship between LC50 values from diet-based bioassays and the larval survivorship in plant-based bioassays. This study provides evidence of Cry3Bb1 resistance throughout the agricultural landscape studied, irrespective of the field history, and highlights the need for improved resistance management approaches, such as better use of integrated pest management to better delay pest resistance.

BACKGROUND Diabrotica virgifera virgifera LeConte is a major pest of corn and causes over a billion dollars of economic loss annually through yield reductions and management costs. Corn producing toxins derived from Bacillus thuringiensis (Bt) has been developed to help manage D. v. virgifera. However, previous studies have demonstrated the ability of this species to evolve resistance to Bt toxins in both laboratory and field settings. RESULTS We used an experimental evolution approach to test the refuge strategies for delaying resistance of D. v. virgifera to corn producing Bt toxin Cry34/35Ab1. In the absence of refuges, D. v. virgifera developed resistance to Bt corn after three generations of selection. In some cases, non‐Bt refuges reduced the level of resistance compared with the strain selected in the absence of refuges, but refuge strains did show reduced susceptibility to Bt corn compared with the unselected strain. CONCLUSIONS In this study, non‐Bt refuges delayed resistance to Bt corn by D. v. virgifera in some cases but not others. Combining the refuge strategy with pyramids of multiple Bt toxins and applying other pest management strategies will likely be necessary to delay resistance of D. v. virgifera to Bt corn. © 2015 Society of Chemical Industry

Background Quantification of adult Aedes aegypti abundance indoors has relied on estimates of relative density (e.g. number of adults per unit of sampling or time), most commonly using traps or timed collections using aspirators. The lack of estimates of the sensitivity of collections and lack of a numerical association between relative and the absolute density of adult Ae. aegypti represent a significant gap in vector surveillance. Here, we describe the use of sequential removal sampling to estimate absolute numbers of indoor resting Ae. aegypti and to calculate calibration coefficients for timed Prokopack aspirator collections in the city of Merida, Yucatan State, Mexico. The study was performed in 200 houses that were selected based on recent occurrence of Aedes -borne viral illness in residents. Removal sampling occurred in 10-minute sampling rounds performed sequentially until no Ae. aegypti adult was collected for 3 hours or over 2 consecutive 10-minute periods. Results A total of 3439 Ae. aegypti were collected. The sensitivity of detection of positive houses in the first sampling round was 82.5% for any adult Ae. aegypti , 78.5% for females, 75.5% for males and 73.3% for blood-fed females. The total number of Ae. aegypti per house was on average ~5 times higher than numbers collected for the first sampling round. There was a positive linear relationship between the relative density of Ae. aegypti collected during the first 10-min round and the absolute density for all adult metrics. Coefficients from the linear regression were used to calibrate numbers from 10-min collections into estimates of absolute indoor Ae. aegypti density for all adults, females and males. Conclusions Exhaustive removal sampling represents a promising method for quantification of absolute indoor Ae. aegypti density, leading to improved entomological estimates of mosquito distribution, a key measure in the assessments of the risk pathogen transmission, disease modeling and the evaluation of vector control interventions.

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