Development, application and evaluation of three novel TaqMan qPCR assays for phosphine resistance monitoring in major stored product pests Tribolium castaneum and Rhyzopertha dominica.

BACKROUND: Stored product protection from insect pests relies heavily on the use of phosphine. The most serious drawback of phosphine is the development of resistance in major stored product insects worldwide, including the red flour beetle, Tribolium castaneum (Herbst) and the lesser grain borer, Rhyzopertha dominica (F.). Two genetic loci are responsible for phosphine resistance: the rph1 (S349G mutation in the cyt-b5-r homolog) in T. castaneum and the rph2 (P45/49S mutation in the dihydrolipoamide dehydrogenase (dld) gene) in T. castaneum and R. dominica. RESULTS: In this study, we have developed and applied high-throughput, practical and specific molecular diagnostics (TaqMan qPCR) for monitoring mutations S349G, P45S and P49S. In our pilot monitoring application, we have included phosphine-resistant and susceptible populations from different parts of the world (USA, Australia, Brazil) and European strains from Greece and Serbia. Our results for the resistant T. castaneum showed a P45S mutant allele frequency (MAF) of 100% and 75.0% in the populations from Serbia and Brazil, respectively. Regarding the susceptible T. castaneum, P45S was detected in Greece (MAF = 62.5%) and was absent in Australia (MAF = 0.0%). Additionally, the S349G mutation was found to be fixed in all resistant populations, while it was also detected in susceptible ones (frequencies: 65.0% and 100.0%). The only case where both mutations were fixed (100%) was a resistant population from Serbia. In R. dominica, the P49S mutation was found only in the two resistant R. dominica populations from Serbia and Greece (50.0% and 100%) and was absent from the susceptible one from Greece; thus, P49S seems to be a satisfactory indicator for monitoring phosphine resistance. CONCLUSIONS: Our P49S detection assay in R. dominica seems to be a viable option in this direction, yet its utilization needs additional large-scale confirmatory work. The identification of additional resistance markers also should be prioritized. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Developing a Hazomalania voyronii Essential Oil Nanoemulsion for the Eco-Friendly Management of Tribolium confusum, Tribolium castaneum and Tenebrio molitor Larvae and Adults on Stored Wheat.

Most insecticides commonly used in storage facilities are synthetic, an issue that generates concerns about food safety and public health. Therefore, the development of eco-friendly pest management tools is urgently needed. In the present study, a 6% (w/w) Hazomalania voyronii essential oil-based nanoemulsion (HvNE) was developed and evaluated for managing Tribolium confusum, T. castaneum, and Tenebrio molitor, as an eco-friendly wheat protectant. Larval and adult mortality was evaluated after 4, 8, and 16 h, and 1, 2, 3, 4, 5, 6, and 7 days, testing two HvNE concentrations (500 ppm and 1000 ppm). T. confusum and T. castaneum adults and T. molitor larvae were tolerant to both concentrations of the HvNE, reaching 13.0%, 18.7%, and 10.3% mortality, respectively, at 1000 ppm after 7 days of exposure. However, testing HvNE at 1000 ppm, the mortality of T. confusum and T. castaneum larvae and T. molitor adults 7 days post-exposure reached 92.1%, 97.4%, and 100.0%, respectively. Overall, the HvNE can be considered as an effective adulticide or larvicide, depending on the target species. Our results highlight the potential of H. voyronii essential oil for developing green nanoinsecticides to be used in real-world conditions against key stored-product pests.

Behavioral Asymmetries Affecting Male Mating Success in Tenebrio molitor (Coleoptera: Tenebrionidae), an Important Edible Species.

The yellow mealworm, Tenebrio molitor L., is one of the most significant insect species of economic importance for producing protein-rich food and feed. The larvae are a promising fishmeal substitute for fish feed, and a good alternative source for human nutrition. In this study, the effect of behavioral asymmetries on male mating success of T. molitor was evaluated. Males performing antennal waving (63%) when detecting a female approached the apex of the female abdomen in a comparable manner from both sides (32% from left side vs 31% from right side). Fewer males showed antennal waving and also raised the anterior part of their body (37%) during mate recognition; 14% of them approached on the apex of female abdomen from the left side, and 23% of them approached from the right side of female body. The duration of mate recognition, antennal tapping, rubbing behavior, and the whole mating sequence of males that moved on the apex of the abdomen from the left side of females was significantly lower over that of males approaching from the right side (10.7, 12.6, 16.4, and 126.1 s for left-biased males vs 12.2, 14.8, 18.6, and 139.6 s for right-biased males, respectively). Concerning mounting side, left-biased males showed a shorter duration of rubbing behavior and the whole mating sequence (15.5 and 123.2 s) over right-biased (18.9 and 138.3 s) and backside approaching males (19.4 and 144.1 s). The duration of mate recognition, antennal tapping, and copula was not affected by the presence of a male laterality bias. Overall, this study sheds light on how laterality affects mating traits and the male success of this important edible insect species.

Insecticidal effect of diatomaceous earth and pirimiphos-methyl against phosphine-susceptible and phosphine-resistant populations of two stored product beetle species.

In the present study, we evaluated the insecticidal efficacy of diatomaceous earth (DE) and pirimiphos-methyl for the control of phosphine-susceptible and phosphine-resistant populations of Tribolium castaneum (Herbst) and Sitophilus oryzae (L.). Insecticides were applied on wheat or rice at two doses: DE was applied at 1000 and 2000 ppm and pirimiphos-methyl at 1 and 5 ppm. Adult mortality was measured after 7, 14, and 21 days of exposure, and progeny production capacity on the treated substrates was evaluated 65 days later. For T. castaneum, we found that DE, at 2000 ppm, was able to provide 100% control of two of the three populations tested, while for the third population mortality reached only 84%. Similarly, there were differences in mortality levels after exposure to DE-treated grains between the two S. oryzae populations tested. At 1 ppm, pirimiphos-methyl was not effective for any of the T. castaneum populations tested, but complete mortality was recorded for all populations at 5 ppm. In general, populations of S. oryzae were more susceptible than those of T. castaneum, for both commodities. Our data indicate that both insecticides can be used with success in phosphine resistance management programs, but there are populations of a given species that may be less susceptible, which constitutes a preliminary screening essential.

One new species and one newly recorded species in the subgenus Myothorax Murray, 1864 from China (Coleoptera: Nitidulidae: Carpophilinae: Carpophilus).

One new species, Carpophilus (Myothorax) brunneus sp. nov., and one newly recorded species, Carpophilus (Myothorax) nepos Murray, 1864, are described or redescribed from China, photographed, and illustrated in detail.

Cool Down-Warm Up: Differential Responses of Stored Product Insects after Gradual Temperature Changes.

Insect survival after exposure to 0 °C for 7 days was examined in laboratory bioassays for control of adults of six major stored-product beetle species, Oryzaephilus surinamensis (L.), the sawtoothed grain beetle, Cryptolestes ferrugineus, (Stephens), the rusty grain beetle, Dermestes maculatus DeGeer, the hide beetle, Sitophilus oryzae (L.), the rice weevil, Tribolium castaneum (Herbst), the red flour beetle, and T. confusum Jacquelin DuVal, the confused flour beetle In this test there were four different acclimation treatments, insects that had been subjected to a pre-acclimation period to 0 °C, a post-acclimation period, both a pre and post-acclimation period, and adults that were not acclimated. Insect survival for all species except S. oryzae was not affected by the exposure to 0 °C, regardless of the acclimation scenario. In contrast, exposure to 0 °C drastically reduced survival of S. oryzae. Moreover, adults that were exposed to the post-acclimation only and un-acclimated adults had lower survival rates than those that had either exposure to pre-acclimation, or to both pre- and post-acclimation. Results of this experiment show that acclimation played a limited role in adult survival of five of the six tested species, and that exposure of adults to 0 °C for 7 d had no effect in survival of these species as well.

Pimpinella anisum essential oil nanoemulsions against Tribolium castaneum-insecticidal activity and mode of action.

The red flour beetle, Tribolium castaneum Herbst (Coleoptera: Tenebrionidae), is an economically important pest of stored products. As possible alternative to conventional insecticides for its management, plant essential oils have gained interest owing to their effectiveness and eco-friendly features. However, they also show some drawbacks, such as low stability, poor water solubility and diffusion, and limited persistence in the environment. A good strategy to overcome these disadvantages is represented by green nanotechnologies. Herein, we developed a nanoemulsion based on the essential oil from Pimpinella anisum L. (Apiaceae) containing 81.2% of (E)-anethole and evaluated its toxicity on T. castaneum adults and F1 progeny, as well as its morphological and histological impact. The aniseed oil nanoemulsion was characterized by the formation of a semi-solid interphase between oil and water; mean drop size was 198.9 nm, PDI was 0.303, zeta potential was - 25.4 ± 4.47 mV, and conductivity was 0.029 mS/cm. The nanoemulsion showed toxicity on T. castaneum (LC50 = 9.3% v/v), with a significant impact on its progeny. Morphological and histological damages triggered by feeding and exposure to the aniseed nanoemulsion were analyzed by scanning electron microscopy (SEM) and light microscopy. Overall, our findings showed that the development of nanoemulsions allows to improve the stability of P. anisum essential oil enhancing its efficacy against stored grain pests and contributing to reduce the use of harmful synthetic insecticides.

Combining electrostatic powder with an insecticide: effect on stored-product beetles and on the commodity.

BACKROUND: The opportunity to reduce the amount of pirimiphos-methyl applied to grain by formulating it in an electrostatic powder was investigated. The insecticidal efficacy of pirimiphos-methyl in EC formulation or formulated using electrostatic powder (EP) as an inert carrier was investigated against Sitophilus oryzae (L.), Oryzaephilus surinamensis (L.), Rhyzopertha dominica (F.) and Tribolium confusum Jacquelin du Val. Furthermore, the adhesive properties of EP to rice, corn and wheat, together with the effect on bulk density and bread- and pasta-making properties, were investigated. RESULTS: The results showed that pirimiphos-methyl formulated with EP provided better efficacy against adults when compared with EC formulation for O. surinamensis and T. confusum, but there was no difference for R. dominica. Progeny production was consistently lower in grain treated with the EP formulation than in grain treated with the EC. Tests showed that EP adhered to the kernels for longer on hard wheat than on maize or rice. In most commodities, EP did not alter the bulk density. Finally, the addition of EP did not affect flour- and bread-making properties, nor the pasta-making properties. CONCLUSIONS: The results of the present study suggest that an EP could be used to reduce the amount of pirimiphos-methyl applied to grain for effective pest control, with no detrimental effects on grain quality. © 2016 Society of Chemical Industry.

Efficacy of Deltamethrin Against Stored-Product Beetles at Short Exposure Intervals or on a Partially Treated Rice Mass.

Stored-product insects can potentially be exposed to grain protectants for variable time periods. Adults of three species, the lesser grain borer, Rhyzopertha dominica (F.) (Coleoptera: Bostrychidae), the granary weevil Sitophilus granarius (L.) (Coleoptera: Curculionidae), and the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) were exposed for 1, 4, 8, and 24 h on brown rice treated with the pyrethroid deltamethrin at the label rate of 0.5 ppm, then removed and placed on untreated rice. Adults of these same species plus the rice weevil, Sitophilus oryzae (L.) (Coleoptera: Curculionidae), the maize weevil, Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae), and the warehouse beetle, Trogoderma variabile Ballion (Coleoptera: Dermestidae) were exposed on treated brown rice mixed with varying amounts of untreated rice to assess progeny production. Immediate and delayed mortality of exposed adults did not exceed 7% for any exposure interval, but progeny production for T. castaneum was generally lower in comparison with that for the other species. Increasing the amount of treated rice decreased progeny production of R. dominica but not for any Sitophilus species. Mixed results were obtained for T. castaneum and T. variabile. Results show that long exposure times and treatment of an entire rice mass may be necessary to give complete control of stored-product beetles.