This study aimed to explore the influence of high voltage electrostatic field (HVEF) treatment on the quorum sensing regulation of Shewanella mediated by Acinetobacter in channel catfish. The research involved the identification of acyl homoserine lactones (AHLs) types within Acinetobacter. The experimental design centered on mixed culture systems comprising radiation-resistant Acinetobacter (AR) by HVEF treatment， alongside Shewanella hafniensis (SH) and Shewanella putrefaciens (SP). A comprehensive analysis was conducted to assess the impact of HVEF treatment on various physiological attributes of Shewanella， including membrane potential， protease activity， biofilm development， extracellular polymeric substance production， cellular aggregation， and motility. In the mixed co-culture system， although strain SP and strain SH could not produce AHLs signaling molecules， they could be promoted by strain AR to improve the rot generating ability and biofilm forming ability. Upon co-cultivation of strain AR with strains SH and SP， a marked enhancement in various indicators was observed: protease activity escalated by 3.29-fold and 4.43-fold， biofilm formation was augmented by 5.00-fold and 4.60-fold， cellular aggregation was bolstered by 1.53-fold and 2.22-fold， swimming motility surged by 5.47-fold and 5.76-fold， and swarming motility was amplified by 4.89-fold and 2.21-fold， respectively. The co-culture system of strain AR treated with HVEF and strain SH and strain SP showed significant effect compared with the group treated without HVEF. The results of this study can provide a theoretical basis for the mechanism of bacterial interaction.