Thermoultrasound-assisted PFB decontamination was optimized for bacteria reduction.
Optimized conditions yielded higher reductions compared with individual treatments.
Predictive models had adequate fitness with temperature variable more significant.
Lipid oxidation and TVB-N after treatment were within limits of fish freshness.
Decontamination was mild on fish microstructure and myofibril degradation.
A novel method of thermoultrasound-assisted plasma functionalized buffer (PFB) for decontaminating grass carp was evaluated using the Box–Behnken design (BBD) with processing variables including PFB generating voltage (P V), ultrasound treatment time (U T) and temperature (T P). The predicted models were found to be significant ( p < 0.05) and displayed sufficient fitness with experimental data as indicated by non-significant ( p > 0.05) lack of fit and high coefficient of determination (R 2 0.97) values. The optimum decontamination conditions for the responses of S. putrefaciens and S. Typhimurium were P V of 66 V, U T of 14.90 min and T P of 60 ℃, achieving reductions of 4.40 and 3.97 log CFU/g, respectively, with a desirability of 0.998. Among the variables, temperature presented higher significance for inactivating bacteria and the production of volatile basic nitrogen and lipid peroxidation under the optimized conditions were within the limits of freshness for grass carp. Additionally, the effects of PFB and the optimized thermoultrasound-assisted PFB decontamination were mild on the microstructure of grass carp with slight ruptures and loose myofibril structures, indicating the potential of thermoultrasound-assisted PFB for seafood products decontamination with reduced processing time.