Fenvalerate is a broadly used type II pyrethroid with a potential toxic effect in fish. However, information on the immunotoxicity of fenvalerate in fish is scarce. Here, to discover the immunotoxicity of fenvalerate and its underlying mechanism in fish, adult Chinese rare minnow was exposed to fenvalerate at 0, 0.3, 1, and 3 μg/L for 28 days and then subjected to Pseudomonas fluorescens (P. fluorescens) challenge. Fenvalerate induced significant pathological changes, with disintegration of cell boundaries in the intestine, epithelial hyperplasia in gills, and vacuolation of hepatocytes at 3 μg/L treatment. Additionally, the pathological characteristics were more serious during P. fluorescens infection after fenvalerate exposure. A significant increase in neutrophil counts was observed after 3 μg/L fenvalerate exposure for 28 days (p < 0.05), whereas significantly increased monocyte and neutrophil counts and greatly decreased lymphocyte counts were detected at 24 h post-injection (hpi) with P. fluorescens (p < 0.05). Furthermore, obvious decreases in LYS, IgM, ALP, and C3 levels were detected in plasma after 3 μg/L fenvalerate exposure for 28 days, which was consistent with the results at 24 and 48 hpi. Notably, fish exposed to fenvalerate suppressed the transcription of TLR-NF-κB signaling pathway-relevant genes in response to P. fluorescens, accompanied by high mortalities and bacterial loads. Therefore, our results demonstrate that fenvalerate at environmentally relevant concentrations caused immunotoxicity in fish. This study highlights the importance of considering the combined effects of chemicals and pathogens to refine our ability to predict the effects of environmental contaminants on aquatic organisms.