Laboratory Selection and Assessment of Resistance Risk in Drosophila suzukii (Diptera: Drosophilidae) to Spinosad and Malathion

Drosophila suzukii (Matsumura) is one of the most economically important pests of soft-skinned fruits worldwide. Repeated insecticide applications commonly used to prevent fruit infestations increase the risk of resistance development in D. suzukii. Assessment of resistance risk in D. suzukii using artificial selection can be valuable in developing proactive resistance management strategies to retain susceptibility in the field populations. Here, we artificially selected a colony of field-collected D. suzukii for resistance against spinosad and malathion. A quantitative genetic approach was then used to estimate realized heritability (h2) of resistance and predict the rates of resistance development. After 10 and 11 generations of selection, resistance to spinosad and malathion in D. suzukii females significantly increased by 7.55- and 2.23-fold, respectively. Based on the predicted rates of resistance development, assuming h2 = 0.14 (mean h2 of spinosad resistance in this study) and 90% of population was killed at each generation, 10-fold increase in LC50 of D. suzukii females would be expected in nine generations for spinosad. However, 10-fold increase in LC50 of D. suzukii females for malathion would be expected in 37 generations, assuming h2 = 0.08 (mean h2 of malathion resistance) and 90% of population was killed at each generation. These results indicate that the risk of resistance in D. suzukii populations exists against both spinosad and malathion. However, resistance would develop faster against spinosad as compared to malathion. Thus, resistance management strategies should be implemented proactively to maintain the effectiveness of these insecticides to control D. suzukii.

Oil emulsion characteristics as significance in efficiency forecast of oil-displacing formulations based on surfactants

Enhanced oil recovery in mature fields can be implemented using chemical flooding with the addition of surfactants using surfactant-polymer (SP) or alkaline-surfactant-polymer (ASP) flooding. Chemical flooding design is implemented taking into account reservoir conditions and composition of reservoir fluids. The surfactant in the oil-displacing formulation allows changing the rock wettability, reducing the interfacial tension, increasing the capillary number, and forming an oil emulsion, which provides a significant increase in the efficiency of oil displacement. The article is devoted with a comprehensive study of the formed emulsion phase as a stage of laboratory selection of surfactant for SP or ASP composition. In this work, the influence of aqueous phase salinity level and the surfactant concentration in the displacing solution on the characteristics of the resulting emulsion was studied. It was shown that, according to the characteristics of the emulsion, it is possible to determine the area of optimal salinity and the range of surfactant concentrations that provide increased oil displacement. The data received show the possibility of predicting the area of effectiveness of ASP and SP formulations based on the characteristics of the resulting emulsion.

The Rapid Evolution of Resistance to Vip3Aa Insecticidal Protein in Mythimna separata (Walker) Is Not Related to Altered Binding to Midgut Receptors

Laboratory selection for resistance of field populations is a well-known and useful tool to understand the potential of insect populations to evolve resistance to insecticides. It provides us with estimates of the frequency of resistance alleles and allows us to study the mechanisms by which insects developed resistance to shed light on the mode of action and optimize resistance management strategies. Here, a field population of Mythimna separata was subjected to laboratory selection with either Vip3Aa, Cry1Ab, or Cry1F insecticidal proteins from Bacillus thuringiensis. The population rapidly evolved resistance to Vip3Aa reaching, after eight generations, a level of >3061-fold resistance, compared with the unselected insects. In contrast, the same population did not respond to selection with Cry1Ab or Cry1F. The Vip3Aa resistant population did not show cross resistance to either Cry1Ab or Cry1F. Radiolabeled Vip3Aa was tested for binding to brush border membrane vesicles from larvae from the susceptible and resistant insects. The results did not show any qualitative or quantitative difference between both insect samples. Our data, along with previous results obtained with other Vip3Aa-resistant populations from other insect species, suggest that altered binding to midgut membrane receptors is not the main mechanism of resistance to Vip3Aa.

Baseline Susceptibility and Laboratory Selection of Resistance to Bt Cry1Ab Protein of Chinese Populations of Yellow Peach Moth, Conogethes punctiferalis (Guenée)

Yellow Peach Moth (YPM), Conogethes punctiferalis (Guenée), is one of the most destructive maize pests in the Huang-Huai-Hai summer maize region of China. Transgenic Bacillus thuringiensis (Bt) maize provides an effective means to control this insect pest in field trials. However, the establishment of Bt resistance to target pests is endangering the continued success of Bt crops. To use Bt maize against YPM, the baseline susceptibility of the local populations in the targeted areas needs to be verified. Diet-overlay bioassay results showed that all the fourteen YPM populations in China are highly susceptible to Cry1Ab. The LC50 values ranged from 0.35 to 2.38 ng/cm2 over the two years of the collection, and the difference between the most susceptible and most tolerant populations was sevenfold. The upper limit of the LC99 estimates of six pooled populations produced >99% larval mortality for representative eight populations collected in 2020 and was designated as diagnostic concentrations for monitoring susceptibility in YPM populations in China. Hence, we evaluated the laboratory selection of resistance in YPM to Cry1Ab using the diet-overlay bioassay method. Although the resistant ratio was generally low, YPM potentially could evolve resistance to Cry1Ab. The potential developmentof resistance by target pests points out the necessity to implement resistance management strategies for delaying the establishment of pest resistance to Bt crops.

Risk Assessment of Flonicamid Resistance in Musca domestica (Diptera: Muscidae): Resistance Monitoring, Inheritance, and Cross-Resistance Potential

Flonicamid is a chordotonal modulator and novel systemic insecticide that has been used frequently for controlling a broad range of insect pests. The risk of flonicamid resistance was assessed through laboratory selection and determining inheritance pattern and cross-resistance potential to five insecticides in house fly, Musca domestica L. Very low to high flonicamid resistance in M. domestica populations was found compared with the susceptible strain (SS). A flonicamid-selected (Flonica-RS) M. domestica strain developed 57.73-fold resistance to flonicamid screened for 20 generations compared with the SS. Overlapping 95% fiducial limits of LC50 of the F1 and F1ǂ, and dominance values (0.87 for F1 and 0.92 for F1ǂ) revealed an autosomal and incomplete dominant flonicamid resistance. The monogenic model of resistance inheritance suggested a polygenic flonicamid resistance. The Flonica-RS strain displayed negative cross-resistance between flonicamid and sulfoxaflor (0.10-fold) or clothianidin (0.50-fold), and very low cross-resistance between flonicamid and flubendiamide (4.71-fold), spinetoram (4.68-fold), or thiamethoxam (2.02-fold) in comparison with the field population. The estimated realized heritability (h2) value of flonicamid resistance was 0.02. With selection mortality 40–90%, the generations required for a 10-fold increase in LC50 of flonicamid were 94–258 at h2 (0.02) and slope (3.29). Flonicamid resistance was inherited as autosomal, incomplete dominant, and polygenic in the Flonica-RS. Negative or very low cross-resistance between flonicamid and sulfoxaflor, clothianidin, flubendiamide, spinetoram, and thiamethoxam means that these insecticides can be used as alternatives for controlling M. domestica. These data can be useful in devising the management for M. domestica.

Evidence for Co-Evolution of Masking With Circadian Phase in Drosophila Melanogaster

Heritable variation in the timing of rhythmic events with respect to daily time cues gives rise to chronotypes. Despite its importance, the mechanisms (clock or non-clock) regulating chronotypes remain elusive. Using artificial laboratory selection for divergent phasing of emergence of adults from pupae, our group has derived populations of Drosophila melanogaster which are early and late chronotypes for eclosion rhythm. Several circadian rhythm characteristics of these populations have since been described. We hypothesized that our selection protocol has inadvertently resulted in selection for masking, a non-clock phenomenon, in the early chronotype due to the placement of our selection window (which includes the lights-ON transition). We designed experiments to discriminate between enhanced masking to light versus circadian clock mediated changes in determining enhanced emergence in the morning window in our early chronotypes. Using a series of phase-shift protocols, LD-DD transition, and T-cycle experiments, we find that our early chronotypes have evolved positive masking, and their apparent entrained phases are largely contributed by masking. Through skeleton T-cycle experiments, we find that in addition to the evolution of greater masking, our early chronotypes have also evolved advanced phase of entrainment. Furthermore, our study systematically outlines experimental approaches to examine relative contributions of clock versus non-clock control of an entrained behavior. Although it has previously been suggested that masking may confer an adaptive advantage to organisms, here we provide experimental evidence for the evolution of masking as a means of phasing that can complement clock control of an entrained behavior.

Evidence for co-evolution of masking and circadian phase in Drosophila melanogaster

Heritable variation in the timing or circadian phases of rhythmic events with respect to daily time cues gives rise to chronotypes. Despite its importance, the mechanisms (clock or non-clock) regulating chronotypes remain elusive. Using artificial laboratory selection for divergent phasing of emergence of adults from pupae, our group has derived populations of Drosophila melanogaster which are early and late chronotypes for eclosion rhythm. Several circadian rhythm characteristics of these populations have since been described. We hypothesized that our selection protocol has inadvertently resulted in selection for masking, a non-clock phenomenon, in the early chronotype due to the placement of our selection window (which includes the lights-ON transition). Based on theoretical predictions and previous studies on our populations, we designed experiments to discriminate between enhanced masking to light versus circadian clock mediated changes in determining enhanced emergence in the morning window in our early chronotypes. Using a series of phase-shift protocols, LD-DD transition, and T-cycle experiments, we find that our early chronotypes have evolved positive masking, and their apparent entrained phases are largely contributed by masking. Through skeleton T-cycle experiments, we find that in addition to the evolution of greater masking, our early chronotypes have also evolved advanced phase of entrainment. Furthermore, our study systematically outlines experimental approaches to examine relative contributions of clock versus non-clock control of an entrained behavior. Although it has previously been suggested that masking may confer an adaptive advantage to organisms, here we provide experimental evidence for the evolution of masking as a mean of phasing of an entrained rhythm that can complement clock control of an entrained behavior.