Under laboratory conditions, the effects of thiamethoxam were investigated in larvae, pupae and emerging honey bees after exposure at larval stages with different concentrations in the food (0.00001 ng/µL, 0.001 ng/µL and 1.44 ng/µL). Thiamethoxam reduced the survival of larvae and pupae and consequently decreased the percentage of emerging honey bees. Thiamethoxam induced important physiological disturbances. It increased acetylcholinesterase (AChE) activity at all developmental stages and increased glutathione-S-transferase (GST) and carboxylesterase para (CaEp) activities at the pupal stages. For midgut alkaline phosphatase (ALP), no activity was detected in pupae stages, and no effect was observed in larvae and emerging bees. We assume that the effects of thiamethoxam on the survival, emergence and physiology of honey bees may affect the development of the colony. These results showed that attention should be paid to the exposure to pesticides during the developmental stages ofthe honey bee. This study represents the first investigation of the effects of thiamethoxam on the development of A. mellifera following larval exposure.

Climate warming is expected to impact the response of species to insecticides. Recent studies show that this interaction between insecticides and temperature can depend on other factors. Here, we tested for the influence of transgenerational effects on the Insecticide × Temperature interaction in the crop pest moth Spodoptera littoralis. Specifically, we analysed reaction norms among experimental clutches based on a split-plot design crossing the factors temperature, insecticide and clutch. The study was performed on 2280 larvae reared at four temperatures (23, 25, 27 and 29 °C), and their response to the insecticide deltamethrin (three concentrations and a control group) was tested. Temperature had a global influence with effects on larval survival, duration of development, pupal body mass, and significant reaction norms of the clutches for temperature variations of only 2 °C. In addition to the expected effect of deltamethrin on mortality, the insecticide slightly delayed the development of S. littoralis, and the effects on mortality and development differed among the clutches. Projection models integrating all the observed responses illustrated the additive effects of deltamethrin and temperature on the population multiplication rate. Variation in the response of the clutches showed that transgenerational effects influenced the impact of insecticide and temperature. Although no evidence indicated that the Insecticide × Temperature interaction depended on transgenerational effects, the studies on the dependence of the Insecticide × Temperature interaction on other factors continue to be crucial to confidently predict the combined effects of insecticides and climate warming.

Under laboratory conditions, the effects of thiamethoxam were investigated in larvae, pupae and emerging honey bees after exposure at larval stages with different concentrations in the food (0.00001 ng/μL, 0.001 ng/μL and 1.44 ng/μL). Thiamethoxam reduced the survival of larvae and pupae and consequently decreased the percentage of emerging honey bees. Thiamethoxam induced important physiological disturbances. It increased acetylcholinesterase (AChE) activity at all developmental stages and increased glutathione-S-transferase (GST) and carboxylesterase para (CaEp) activities at the pupal stages. For midgut alkaline phosphatase (ALP), no activity was detected in pupae stages, and no effect was observed in larvae and emerging bees. We assume that the effects of thiamethoxam on the survival, emergence and physiology of honey bees may affect the development of the colony. These results showed that attention should be paid to the exposure to pesticides during the developmental stages of the honey bee. This study represents the first investigation of the effects of thiamethoxam on the development of A. mellifera following larval exposure.

Morphological tools can assist in the evaluation of effects of insecticides on non-target insects. Pyriproxyfen, a juvenile hormone analog, is known to interfere with growth and metamorphosis of insects. However, there are studies showing indirect effects on natural enemies, including green lacewings. Few prior studies describe morphological effects of pyriproxyfen on target insect organs, especially on natural enemies. Through morphological tools, this study aimed to characterize the midgut and fat body, both important organs of digestion and great metabolic activity respectively, of the predator Ceraeochrysa claveri after chronic exposure to pyriproxyfen. Larvae of C. claveri were fed Diatraea saccharalis egg clusters treated with pyriproxyfen in solution of 50 or 100 mg a.i. L⁻¹ throughout the larval stage. The biological data revealed significant increases in development time, especially in the third instar, and in cumulative mortality from the prepupal into the pupal stage. Morphological analysis of adult midgut (≤24 h old) showed damage including formation of epithelial folds, intercellular spaces, emission of cytoplasmic protrusions. Both fat body regions presented decrease of lipid droplets, vacuolization of trophocytes and mitochondrial injury featuring a multisystemic action. In both organs, pyriproxyfen exposure induced significant oxidative stress by mitochondrial superoxide production. Cytoprotective responses were induced in midgut and fat body cells by augmenting the number of cytoplasmic granules containing calcium and expression of HSP 90. Both organs proved to be efficient in presenting histopathological alterations, showing the sensitivity and applicability of this morphological tool for evaluating other insecticides in non-target organisms.

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.

In most cases, honey bees experience pesticide pollution in a long-term period through direct or indirect exposure, such as the development process from larvae to the pre-harvest stage. At present, little is known about how honey bees respond to pesticide stresses during the continuous development period. This study aims to examine effects of long-term acetamiprid exposure on the development and survival of honey bees, and further present the expression profile in larvae, 1-day-old, and 7-day-old adult worker bees that related to immune, detoxification, acetylcholinesterase (AChE) and memory. Honey bees from 2-day-old larvae to 14-day-old adults except the pupal stage were continuously fed with different acetamiprid solutions (0, 5, and 25 mg/L). We found that acetamiprid over 5 mg/L disturbed the development involving birth weight and emergence rate of newly emerged bees, and reduced the proportion of capped cells of larvae at 25 mg/L; gene expression related to immune and detoxification of worker bees exposed to acetamiprid was roughly activated, returned and then inhibited from larval to emerged and to the late adult stage, respectively. Moreover, lifespans of bees treated with acetamiprid at 25 mg/L were significantly reduced. The present study reflects the potential risk for honey bees continuously exposed to acetamiprid in the development stage.

Uptake of Cry toxins by insect natural enemies has rarely been considered and bioaccumulation has not yet been demonstrated. Uptake can be demonstrated by the continued presence of Cry toxin after exposure has stopped and gut contents eliminated. Bioaccumulation can be demonstrated by showing uptake and that the concentration of Cry toxin in the natural enemy exceeds that in its food. We exposed larvae of the aphidophagous predator, Harmonia axyridis, to Cry1Ac and Cry1F through uniform and constant tritrophic exposure via an aphid, Myzus persicae, and looked for toxin presence in the pupae. We repeated the experiment using only Cry1F and tested newly emerged adults. Both Cry toxins were detected in pupae, and Cry1F was detected in recently emerged, unfed adults. Cry1Ac was present 2.05 times and Cry1F 3.09 times higher in predator pupae than in the aphid prey. Uptake and bioaccumulation in the third trophic level might increase the persistence of Cry toxins in the food web and mediate new exposure routes to natural enemies.

The objective of this study was to evaluate the biological and nutritional characteristics of Spodoptera frugiperda (Lepidoptera: Noctuidae), an arthropod pest widely distributed in agricultural regions, after exposure to nano-CeO₂ via an artificial diet and to investigate the presence of cerium in the body of this insect through X-ray fluorescence mapping. Nano-CeO₂, micro-CeO₂, and Ce(NO₃)₃ were incorporated into the diet (0.1, 1, 10, and 100 mg of Ce L⁻¹). Cerium was detected in caterpillars fed with diets containing nano-CeO₂ (1, 10 and 100 mg of Ce L⁻¹), micro-CeO₂ and Ce(NO₃)₃, and in feces of caterpillars from the first generation fed diets with nano-CeO₂ at 100 mg of Ce L⁻¹ as well. The results indicate that nano-CeO₂ caused negative effects on S. frugiperda. After it was consumed by the caterpillars, the nano-CeO₂ reduced up to 4.8% of the pupal weight and 60% of egg viability. Unlike what occurred with micro-CeO₂ and Ce(NO₃)₃, nano-CeO₂ negatively affected nutritional parameters of this insect, as consumption rate two times higher, increase of up to 80.8% of relative metabolic rate, reduction of up to 42.3% efficiency of conversion of ingested and 47.2% of digested food, and increase of up to 1.7% of metabolic cost and 8.7% of apparent digestibility. Cerium caused 6.8–16.9% pupal weight reduction in second generation specimens, even without the caterpillars having contact with the cerium via artificial diet. The results show the importance of new ecotoxicological studies with nano-CeO₂ for S. frugiperda in semi-field and field conditions to confirm the toxicity.

The use of insecticides based on metabolites found in live organisms, such as the insecticide spinosad, has been an option for the control of agricultural pests because of the allegedly low toxicological risk for nontarget arthropods, such as stingless bees. In the current study, we evaluate the effects of chronic oral exposure to spinosad during the larval phase on survival, developmental time, body mass, midgut epithelial remodeling, and the peritrophic matrix (PM) of Partamona helleri stingless bee workers. Worker larvae that were raised in the laboratory were orally exposed to different concentrations (0, 6.53, 13.06, 32.64, and 3,264 ng. a.i. bee⁻¹) of spinosad (formulation), and the resulting survival, developmental time, and body mass were studied. The concentration of spinosad recommended for use in the field (3,264 ng. a.i. bee⁻¹) reduced the survival of workers during development. Also, sublethal concentrations of spinosad delayed the development and caused morphological changes in the midgut epithelium. Finally, the chronic exposure of larvae to 32.64 ng. a.i. bee⁻¹ spinosad also altered the remodeling of the midgut during metamorphosis and affected the organization of the PM of larvae, pupae, and adults. Our data suggest possible environmental risks for using spinosad in cultures that are naturally pollinated by stingless bees.

Low-density polyethylene (LDPE), biaxially oriented polypropylene (BOPP), and expanded polystyrene (EXPS) are the most common plastics found in every home of the world, but only ~ 10% enter the recycling chains. Consequently, the study of plastic biodegradation by microorganisms and insects, such as the wax moths, has gained special interest. Galleria mellonella (L.) has been shown to consume single-layered polyethylene and polystyrene, though biological impacts of this consumption have been rarely reported. We evaluated the consumption of different plastics by G. mellonella larvae (L7, mean size: 25–30 mm) and its effect on larval duration, survival, and development. For this, we offered the larvae five diets: single-layered LDPE, EXPS, BOPP, triple-layered polyethylene (SB, for silo-bags), and a control with beeswax. We recorded the state and weight of the materials and the state of larvae until they reached the adult stage. Larvae consumed more PE (both LDPE and SB) and EXPS than BOPP; still, they were able to emerge as adults in all treatments. Larvae that consumed plastics turned into pupal stage faster than those that consumed beeswax, regardless of the type and amount of plastic consumed. This is the first report of wild G. mellonella larvae in Argentina consuming biaxially polypropylene and silo-bags.