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 LC₅₀ value of 216.3 mg a.i. L⁻¹) than CYP (96-h LC₅₀ 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.

农药暴露仍然是传粉昆虫数量下降的主要因素之一。迫切需要评估混合物对传粉媒介风险评估的影响，因为它们经常接触多种农用化学品。在本研究中，我们探讨了噻虫啉 (THI) 和环丙唑醇 (CYP) 对蜜蜂 ( Apis mellifera L.) 的混合毒性作用。我们的研究结果表明，THI 对意大利蜜蜂的急性毒性（96 小时 LC ₅₀值为 216.3 mg ai L ⁻¹）高于 CYP（96 小时 LC ₅₀值为 601.4 mg ai L ^{−1 ）}). 值得注意的是，THI 和 CYP 的混合物对蜜蜂产生了强烈的协同作用。同时检测解毒酶细胞色素P450s（CYP450s）和神经靶酶乙酰胆碱酯酶（AChE）的活性，以及​​7个基因（CRBXase、CYP306A1、CYP6AS14、apidaecin、defensing - 2、vtg、gp- 93) 与解毒代谢、免疫反应、发育和内质网应激相关，与 THI 或 CYP 的相应个体暴露相比，在联合治疗中显着改变。这些数据表明 THI 和 CYP 的混合物可能会扰乱蜜蜂的生理稳态。本研究为深入研究农药混配对蜜蜂健康的影响提供了理论依据。我们的研究也为农业生产中农药混剂的合理施用有效保护传粉媒介提供了重要指导。