BackgroundProbiotics have excellent antioxidant and antibacterial effects and are also considered to be promising natural biological preservatives. This study examined the freeze–thaw stability of the antioxidant and antibacterial activity of probiotic Bacillus velezensis PJP10, and evaluated the effect of different concentrations of PJP10 (106, 107, and 108 CFU mL−1) on microbial growth, textural characteristics, physicochemical properties, and lipid oxidation of grass carp fillets during repeated freeze–thaw cycles.ResultsFreeze–thaw cycles had little effect on the antioxidant and antibacterial stability of PJP10. Treatment with the PJP10 strain slowed effectively the increase in Aeromonas spp. counts (ASC), pH, total volatile basal nitrogen (TVB‐N), and thiobarbituric acid (TBA) values of the freeze–thaw fish fillets. It maintained the textural quality of fillets, inhibiting the reduction of hardness, springiness, gumminess, cohesiveness, chewiness, and resilience values, and reduced lipid oxidation, as evidenced by decreased carbonyl values and conjugated diene values after multiple freeze–thaw cycles.ConclusionProbiotic B. velezensis PJP10 had protective effects on microbiological, textural, and physicochemical properties, and lipid oxidation of grass carp fillets during repeated freeze–thaw storage, suggesting a potential application of this strain in the freeze–thaw preservation of freshwater fish. © 2024 Society of Chemical Industry.