Biosolids are waste from wastewater treatment and have a high content of organic matter and nutrients. In this study, not conventional physicochemical and biological properties of biosolids produced during different seasons of the year were evaluated. These properties are not considered in environmental regulations; however, they are of agronomic interest as indicators of quality and functionality in soils. Also, molecular analysis by Fourier-transform infrared (FTIR) was conducted, enzymatic analysis using the APIZYM^® system was performed and two indices of functional and microbial diversity were established. The results showed that the biosolids had a high content of total organic carbon, total nitrogen, P, and K. FTIR analyses showed that chemical composition of biosolids was similar during all year. The C and N of microbial biomass demonstrated presence of active microorganisms, as well as a uniformity in its richness and abundance of species that could present a positive synergy with soil microorganisms. The enzymatic activities showed that the biosolids contained an enzymatic machinery available to promote the mineralisation of the organic matter of biosolids and could even transcend into the soil. Finally, biological properties can be used as indicators of quality and functionality of biosolids before being used as an organic amendment, especially in agricultural soils.