The emergence of antimicrobial-resistant bacteria in aquaculture has raised the need for alternative strategies to control fish infections. Antimicrobial-producing bacteria have been explored as probiotics or biocontrol agents, but their mechanisms of action and impact on host-associated microbiota remain poorly understood. Here, we identified Pseudomonas mosselii KH-ZF1, a bacterium isolated from fish epidermal mucus, as a producer of antimicrobial substances. When applied to zebrafish, KH-ZF1 transiently adhered to the epidermal mucus and altered the composition of the skin microbiota. Under an appropriate administration condition, KH-ZF1 treatment significantly improved survival in zebrafish infected with Yersinia ruckeri, and suppressed pathogen growth on the skin surface. However, in the absence of KH-ZF1 or inappropriate conditions, Y. ruckeri dominated the epidermal bacterial community. The antimicrobial compound produced by KH-ZF1 was identified as Fluviol C (FluC), a pigmented metabolite previously reported from Pseudomonas fluorescens. FluC inhibited the growth of multiple fish pathogens at low concentrations, but exhibited toxicity to zebrafish even below its minimum inhibitory concentration. Intriguingly, FluC at sub-inhibitory levels induced bacterial substitution in the epidermal microbiota, mimicking the effects of KH-ZF1. These findings demonstrate that KH-ZF1 alters host resistance to infection by promoting bacterial substitution on the fish skin by producing FluC. Our study highlights a microbiota-mediated mechanism by which antimicrobial-producing bacteria can control infection through fish epidermis, offering a potential alternative to traditional antibiotics in aquaculture.