Insecticidal Cry proteins from Bacillus thuringiensis (Bt) can be transferred from genetically engineered crops to herbivores to natural enemies. For the lady beetle Harmonia axyridis, we investigated potential uptake of Cry proteins from the gut to the body and intergenerational transfer. Third and fourth instar H. axyridis fed with pollen or spider mites from SmartStax maize contained substantial amounts of Cry1A.105, Cry1F, Cry2Ab2, Cry3Bb1, and Cry34Ab1. Cry protein concentrations in lady beetle larvae were typically one order of magnitude lower than in the food. When H. axyridis larvae were fed Bt maize pollen, median amounts of Cry protein in the non-feeding pupae were below the limit of detection except for small amounts of Cry34Ab1. No Cry protein was detected in pupae when spider mites were used as food. Cry protein concentrations decreased quickly after H. axyridis larvae were transferred from pollen or spider mites to Bt-free food. Aphids contained very low or no detectable Cry protein, and no Cry protein was found in H. axyridis larvae fed with aphids, and in pupae. When H. axyridis adults were fed with Bt maize pollen (mixed with Ephestia kuehniella eggs), the median concentrations of Cry proteins in lady beetle eggs were below the limit of detection except for Cry34Ab1 in eggs laid later in adult life. No Bt protein was detected in eggs laid by H. axyridis females fed with aphids from Bt maize. Our results confirm previous observations that Cry proteins are degraded and excreted quickly in the arthropod food web without evidence for bioaccumulation. Despite the fact that small amounts of Cry proteins were detected in some samples of the non-feeding pupal stage of H. axyridis as well as in eggs, we conclude that this route of exposure is unlikely to be significant for predators or parasitoids in a Bt maize field.

Etofenprox is a broad spectrum pyrethroid insecticide with low toxicity to mammals, fishes, and honeybees. In the present study it was evaluated as grain protectant against Ephestia kuehniella (Lepidoptera: Pyralidae) larvae, Prostephanus truncatus (Coleoptera: Bostrychidae) adults, Rhyzopertha dominica (Coleoptera: Bostrychidae) adults, Sitophilus oryzae (Coleoptera: Curculionidae) adults, and Tribolium confusum (Coleoptera: Tenebrionidae) larvae and adults. Etofenprox was applied at 0.1, 1, 5, and 10 ppm on wheat, or maize in the case of P. truncatus, and tested at different combinations of temperatures (20, 25, and 30 °C) and relative humidity (RH) levels (55 and 75%). Progeny production of the tested coleopteran adult species was also assessed. For E. kuehniella, after 21 days of exposure 75.6% of the exposed larvae were killed at 20 and 30 °C/55% RH. Mortality of P. truncatus adults reached 99.4 and 97.8% at 10 ppm of 55 and 75% RH, respectively, at 30 °C. For S. oryzae, after 21 days of exposure, mortality was moderate at both RH levels, even at the elevate doses, reaching 66.7% at 10 ppm at 20°C/75% RH. All R. dominica adults died 21 days post-exposure at 30°C/55% RH and 25 or 30°C/75% RH at 10 ppm. For T. confusum adults, mortality was 81.1% 10 ppm 21 days post-exposure at 20°C/75% RH. Etofenprox killed 99.4% of the exposed T. confusum larvae at 10 ppm respectively 14 days post-exposure at 25°C/55% RH. Concerning progeny production, complete suppression was recorded for P. truncatus, R. dominica, and T. confusum in various combinations of temperature/RH. Our findings indicate that etofenprox is a well-promising insecticide for the protection of stored grains. However, its performance differs among insect species and abiotic conditions.

Several pyrrole derivatives exhibit insecticidal activity and can be effective as grain protectants. In the present study, we evaluate the insecticidal efficacy of six novel pyrrole derivatives, namely methyl 3-(methylthio)-4,6-dioxo-5-phenyl-1,3a,4,5,6,6a-hexahydropyrrolo[3,4-c]pyrrole-1 carboxylate (compound syn) (2a-syn), methyl 3-(methylthio)-4,6-dioxo-5-phenyl-1,3a,4,5,6,6a-hexahydropyrrolo[3,4-c]pyrrole-1-carboxylate (compound anti) (2a-anti), methyl 3-(benzylthio)-4,6-dioxo-5-phenyl-1,3a,4,5,6,6a-hexahydropyrrolo[3,4-c]pyrrole-1-carboxylate (compound syn) (2f-syn), methyl 3-(benzylthio)-4,6-dioxo-5-phenyl-1,3a,4,5,6,6a-hexahydropyrrolo[3,4-c]pyrrole-1-carboxylate (compound anti) (2f-anti), methyl 3-(butylthio)-4,6-dioxo-5-phenyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrrole-1-carboxylate (3e), and methyl 2-benzyl-3-(methylthio)-4,6-dioxo-5-phenyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrrole-1-carboxylate (0665), against four important species infesting stored products, the rice weevil, Sitophilus oryzae (Coleoptera: Curculionidae), the lesser grain borer, Rhyzopertha dominica (Coleoptera: Bostrychidae), the confused flour beetle, Tribolium confusum (Coleoptera: Tenebrionidae), and the Mediterranean flour moth, Ephestia kuehniella (Lepidoptera: Pyralidae). The six pyrrole derivatives were evaluated on wheat at different doses (0.1, 1, and 10 ppm) and exposure intervals (7, 14, and 21 days). For S. oryzae adults, the highest mortality was recorded at 10 ppm of 2a-syn (36.7%) followed by 2f-syn (32.2%) and 2f-anti (27.8%) after 21 days of exposure. Regarding progeny production, the application of the six pyrrole derivatives significantly reduced offspring emergence if compared with the controls. After 21 days, mortality of R. dominica reached 50% testing 10 ppm of 2f-syn, followed by 2a-syn (46.7%), 2f-anti (41.1%), and 2a-anti (33.3%), while for 3e and 0665, mortality remained low, not exceeding 17.8%. Mortality of T. confusum adults was very low, ranging from 0 to 16.7% after 21 days of exposure. Progeny production was low (< 1.7 individuals per vial) for all doses of the tested pyrrole derivatives, including control vials. For 2a-syn, 2a-anti, 2f-anti, and 0665, no progeny production was recorded testing 1 and 10 ppm, while for 2f-syn and 3e, no offspring emergence was noted testing 10 ppm. For T. confusum larvae, after 21 days of exposure, mortality reached 62.2% testing 10 ppm of 3e followed by 0665 (55.6%) and 2a-anti (42.2%). For E. kuehniella larvae, mortality reached 57.8% at 10 ppm of 2a-syn, followed by the pyrrole derivative 2f-anti (43.3%) after 21 days of exposure. Overall, these results show that the efficacy of pyrrole derivatives strongly varied according to the exposure interval, tested dose, treated insect species and developmental instar. The tested pyrrole derivatives, with special reference to 2a-syn, 2a-anti, 2f-syn, 2f-anti and 0665, are slow-acting compounds exerting relevant toxicity on key stored-product pests over time. They can be considered further for assays with selected blends aiming to develop novel control tools against stored-product pests in real-world conditions.