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Pesticide exposure remains one of the main factors in the population decline of insect pollinators. It is urgently necessary to assess the effects of mixtures on pollinator risk assessments because they are often exposed to numerous agrochemicals. In the present study, we explored the mixture toxic effects of thiacloprid (THI) and cyproconazole (CYP) on honey bees (Apis mellifera L.). Our findings revealed that THI possessed higher acute toxicity to A. mellifera (96-h LC 50 value of 216.3 mg a.i. L -1 ) than CYP (96-h LC 50 value of 601.4 mg a.i. L -1 ). It's worth noting that the mixture of THI and CYP exerted an acute synergistic effect on honey bees. At the same time, the activities of detoxification enzyme cytochrome P450s (CYP450s) and neuro target enzyme Acetylcholinesterase (AChE), as well as the expressions of seven genes (CRBXase, CYP306A1, CYP6AS14, apidaecin, defensing-2, vtg, and gp-93) associated with detoxification metabolism, immune response, development, and endoplasmic reticulum stress, were significantly altered in the combined treatment compared with the corresponding individual exposures of THI or CYP. These data indicated that a mixture of THI and CYP could disturb the physiological homeostasis of honey bees. Our study provides a theoretical basis for in-depth studies on the impacts of pesticide mixtures on the health of honey bees. Our study also provides important guidance for the rational application of pesticide mixtures to protect pollinators in agricultural production effectively., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)

The extensive use of pesticides has negative effects on the health of insect pollinators. Although pollinators in the field are seldom exposed to individual pesticides, few reports have assessed the toxic impacts of pesticide combinations on them. In this work, we purposed to reveal the combined impacts of tetrachlorantraniliprole (TET) and tebuconazole (TEB) on honey bees (Apis mellifera L.). Our data exhibited that TET had greater toxicity to A. mellifera (96-h LC 50 value of 298.2 mg a.i. L -1 ) than TEB (96-h LC 50 value of 1,841 mg a.i. L -1 ). The mixture of TET and TEB displayed acute synergistic toxicity to the pollinators. Meanwhile, the activities of CarE, CYP450, trypsin, and sucrase, as well as the expressions of five genes (ppo, abaecin, cat, CYP4G11, and CYP6AS14) associated with immune response, oxidative stress, and detoxification metabolism, were conspicuously altered when exposed to the mixture relative to the individual exposures. These results provided an overall comprehension of honey bees upon the challenge of sublethal toxicity between neonicotinoid insecticides and triazole fungicides and could be used to assess the intricate toxic mechanisms in honey bees when exposed to pesticide mixtures. Additionally, these results might guide pesticide regulation strategies to enhance the honey bee populations., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Insect pollinators are routinely exposed to a complex mixture of many pesticides. However, traditional environmental risk assessment is only carried out based on ecotoxicological data of single substances. In this context, we aimed to explore the potential effects when worker honey bees (Apis mellifera L.) were simultaneously challenged by thiamethoxam (TMX) and flusilazole (FSZ). Results displayed that TMX possessed higher toxicity to A. mellifera (96-h LC 50 value of 0.11 mg a. i. L -1 ) than FSZ (96-h LC 50 value of 738 mg a. i. L -1 ). Furthermore, the mixture of TMX and FSZ exhibited an acute synergistic impact on the pollinators. Meanwhile, the activities of SOD, caspase 3, caspase 9, and PPO, as well as the expressions of six genes (abaecin, dorsal-2, defensin-2, vtg, caspase-1, and CYP6AS14) associated with oxidative stress, immune response, lifespan, cell apoptosis, and detoxification metabolism were noteworthily varied in the individual and mixture challenges than at the baseline level. These data revealed that it is imminently essential to investigate the combined toxicity of pesticides since the toxicity evaluation from individual compounds toward honey bees may underestimate the toxicity in realistic conditions. Overall, the present results could help understand the potential contribution of pesticide mixtures to the decline of bee populations., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Although pesticides commonly exist as combinations in real-life situations of the aquatic ecosystem, the impact of the toxicity of their mixtures has remained largely unclear. In this study, we investigated the combined effects of two neurotoxic pesticides, including one organophosphate insecticide phoxim (PHO) and one pyrethroid insecticide lambda-cyhalothrin (LCY), on the embryos of the small yellow croaker (Larimichthys polyactis), and their potential pathways. LCY exhibited higher toxicity relative to PHO, with a 72-h LC 50 value of 0.0074 mg a.i. L -1 , while the corresponding value for PHO was 0.12 mg a.i. L -1 . The mixture of PHO and LCY exerted a synergistic effect on the embryos of L. polyactis. The activities of antioxidant enzyme CAT and apoptotic enzyme caspase 3 were substantially changed in most single and combined exposure groups relative to the baseline value. Under both single and combined exposures, more significant changes were found in the mRNA expression of five genes, including the immunosuppression gene ngln2, the apoptosis gene P53, the endocrine system gene cyp19a1b, as well as neurodevelopment genes of ap and acp2, relative to the baseline value. Furthermore, the non-target metabolomic analysis demonstrated that hundreds of differential metabolites, including two bile acids (taurodeoxycholic acid and tauroursodeoxycholic acid), were significantly increased in the exposure groups. The bile acids were closely associated with the gut microbiota, and 16S rRNA sequencing results demonstrated dysfunction of the gut microbiota after exposure, especially in the combined exposure group. Our findings indicated that there might be a potential risk connected to the co-occurrence of these two pesticides in aquatic vertebrates. Consequently, future ecological risk assessments should incorporate synergistic mixtures because the current risk assessments do not consider them., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Ltd. All rights reserved.)