Abstract To better understand the role of fermentative microorganisms in lipid changes during the processing of fermented fish, sterile fish paste with or without the antioxidant tert-butylhydroquinone (TBHQ) was used to investigate the roles of three strains isolated from traditional fermented fish in free fatty acid (FFA) liberation and lipid oxidation. The results showed that the addition of 0.03% TBHQ had no significant influence (P > 0.05) on the strain growth but could effectively inhibit fatty acid oxidation. Staphylococcus xylosus 135 demonstrated the highest liberation of FFAs, followed by Saccharomyces cerevisiae 31, with its released FFAs being almost all polyunsaturated fatty acids (PUFA). Furthermore, the highest amount of PUFA oxidation in FFAs was observed in the samples inoculated with Staphylococcus xylosus 135 (802 mg/kg DM), followed by Lactobacillus plantarum 120 (476 mg/kg DM) and Saccharomyces cerevisiae 31(388 mg/kg DM). The results suggested that Staphylococcus xylosus may play a major role in the lipolysis and lipid oxidation during the processing of fermented fish. Lactobacillus plantarum also contributed to the fatty acid oxidation although it had no obvious lipolysis ability. Highlights • Role of microbes in free fatty acids release and oxidation was determined. • 0.03% TBHQ had no effect on strain growth but inhibit fatty acids oxidation. • Oxidation caused the major loss of fatty acids during fermentation of fish. • Staphylococcus xylosus 135 had the highest ability of lipolysis and lipid oxidation. • Lactobacillus plantarum 120 contributed to lipid oxidation but had no lipolytic activity. [ABSTRACT FROM AUTHOR]