In this work the thermal diffusivity (D) of Astyanax lacustris fish scale is investigated aiming to use it for environmental integrity certification. The D values were obtained by a relatively simple procedure by a photoacoustic method. The chosen fish species is from wide occurrence in Brasil's basins. It has short migration, and it has also been used as environmental bioindicator. The results obtained in 195 scales sampled from three different streams in the Midwest region in Brazil gives an average value of D ~ 4 × 10−3 cm2/s. ANCOVA analysis demonstrated that D values are able to differentiate among the three basins and indicates that it is dependent on the scales thickness and water conductivity. This last one is strongly affected by biotic and abiotic actions, so that D values measured by photoacoustic method can be used for interpreting the environmental integrity from where the fishes were sampled. Display omitted •A relatively simple procedure by PAS to determine the thermal diffusivity of fish scales in an optically transparent range•ANCOVA suggests the thermal diffusivity of fish scales as a new parameter for environmental integrity monitoring•The thermal diffusivity of the fish scales are related to their thickness