Vibrio campbellii is a significant pathogen in shrimp aquaculture, causing luminous vibriosis and leading to considerable declines in productivity and quality. The rapid emergence of multi-drug and detergent-resistant strains presents a major challenge in controlling this pathogen. This study investigates the inhibitory effects of rambutan peel extract (RPE) on quorum sensing (QS) systems, biofilm formation, and virulence-related traits in V. campbellii. The minimum inhibitory concentration (MIC) of RPE was found to be 2048 μg/ml for the pathogenic strain V. campbellii HY01 and 1024 μg/ml for the non-pathogenic QS reporter strains. Sub-inhibitory concentrations significantly reduced bioluminescence in V. campbellii, indicating interference with QS systems, particularly harveyi autoinducer-1 (HAI-1) and autoinducer-2 (AI-2). RPE disrupted autoinducer detection, down-regulated the expression of QS sensor genes, inhibited phosphorylation, and affected the transcription of QS regulator AphA. Additionally, RPE reduced biofilm formation, swimming motility, caseinase production, and virulence gene expression in the shrimp pathogenic strain HY01. These findings demonstrate the strong anti-QS activity of RPE against V. campbellii by targeting QS systems, phosphorylation pathways, and the master QS regulator. The study highlights the potential of RPE as a sustainable approach to control luminous vibriosis, offering a promising strategy for managing disease outbreaks and improving shrimp health in aquaculture.