The physicochemical stability of six bread formulations with different tilapia flour (BTF) levels (0%, 2.5%, 5%, 10%, 15%, and 20%) in substitution to wheat flour was investigated regarding moisture content, water activity (a w ), pH, instrumental color parameters, texture profile, lipid and protein oxidation on days 0, 2, 4, 6 and 8 at 25°C. BTF10%, BTF15%, and BTF20% displayed lower (P < 0.05) moisture, aw, lightness index, cohesiveness, springiness and resilience, and higher (P < 0.05) pH, redness index, yellowness index, hardness, chewiness, lipid and protein oxidation compared to BTF0%, BTF2.5%, and BTF5%. Nevertheless, pH drops, lipid and protein oxidation were less pronounced (P < 0.05) for BTF10%, BTF15%, and BTF20% during storage. Although quality loss has been observed in breads containing ≥ 10% TF, bread at 5% TF did not display altered traditional wheat bread physicochemical characteristics and may be an attractive alternative for the health food market.