Acetamiprid is the only neonicotinoid registered in the European Union because the risks of neonicotinoids to honey bees and other pollinators are strictly regulated. Herein, we orally exposed honey bee colonies to sublethal concentrations of acetamiprid (20 μg/L) under isolated conditions. After one month of continuous exposure, the emerging bees and queens were collected and analyzed via high-throughput label-free quantitative proteomics using a data-independent acquisition strategy. Six and 34 significantly differentially expressed proteins (DEPs) were identified in the emerging bees and queens, respectively. Mrjp3 was the only DEP found in both sample types/castes, and its opposite regulation illustrated a differential response. The DEPs in the emerging bees (H/ACA RNP, Rap1GAP, Mrjp3, and JHE) suggested that sublethal exposure to acetamiprid affected cell cycle-related signaling, which may affect the life history of workers in the colony. The DEPs with increased levels in queens, such as Mrjps 1–4 and 6–7, hymenoptaecin, and apidaecin 22, indicated an activated immune response. Additionally, the level of farnesyl pyrophosphate synthase (FPPS), which is essential for the mevalonate pathway and juvenile hormone biosynthesis, was significantly decreased in queens. The impaired utilization of juvenile hormone in queens supported the identification of additional DEPs. Furthermore, the proteome changes suggested the existence of increased neonicotinoid detoxification by UDP-glucuronosyltransferase and increased amino acid metabolism. The results suggest that the continuous exposure of bee colonies to acetamiprid at low doses (nanograms per gram in feed) may pose a threat to the colonies. The different exposure routes and durations for the emerging bees and queens in our experiment must be considered, i.e., the emerging bees were exposed as larvae via feeding royal jelly and beebread provided by workers (nurse bees), whereas the queens were fed royal jelly throughout the experiment. The biological consequences of the proteomic changes resulting from sublethal/chronic exposure require future determination.

中文翻译：

---

亚致死的啶虫脒暴露会诱导免疫力，抑制蜂王中与保幼激素合成相关的途径，并影响新出现的蜜蜂的周期相关信号传导

啶虫脒是唯一在欧盟注册的新烟碱类杀虫剂，因为新烟碱类杀虫剂对蜜蜂和其他传粉媒介的风险受到严格监管。在此，我们在隔离条件下将蜜蜂群体口服暴露于亚致死浓度的啶虫脒（20 μg/L）。经过一个月的连续暴露后，使用数据独立的采集策略，通过高通量无标记定量蛋白质组学收集和分析新出现的蜜蜂和蜂王。在新出现的蜜蜂和蜂王中分别鉴定出 6 个和 34 个显着差异表达蛋白 (DEP)。 Mrjp3 是在两种样本类型/种姓中发现的唯一 DEP，其相反的调节说明了不同的反应。新兴蜜蜂中的 DEP（H/ACA RNP、Rap1GAP、Mrjp3 和 JHE）表明亚致死暴露于啶虫脒会影响细胞周期相关信号传导，这可能会影响蜂群中工蜂的生活史。蜂王中 DEP 水平升高，例如 Mrjps 1-4 和 6-7、hymenoptaecin 和 apidaecin 22，表明免疫反应被激活。此外，蜂王中对甲羟戊酸途径和保幼激素生物合成至关重要的法呢基焦磷酸合酶（FPPS）水平显着降低。蜂王对保幼激素的利用受损支持了额外 DEP 的鉴定。此外，蛋白质组的变化表明UDP-葡萄糖醛酸基转移酶对新烟碱类解毒作用的增强以及氨基酸代谢的增强。结果表明，蜂群持续接触低剂量（饲料中每克纳克）啶虫脒可能会对蜂群造成威胁。在我们的实验中，必须考虑新出现的蜜蜂和蜂王的不同暴露途径和持续时间，即新出现的蜜蜂在幼虫时期通过喂养工蜂（护士蜂）提供的蜂王浆和蜂面包而暴露，而蜂王则始终喂食蜂王浆本实验。亚致死/慢性暴露引起的蛋白质组变化的生物学后果需要未来确定。