Biochar amendment is becoming a promising technology for mining soil restoration. The addition of biochar can improve soil microbiological parameters related to soil quality, such as enzyme activities. The aim of the present research was to evaluate the effect of rabbit manure (RM) and two rabbit manure biochars prepared at two pyrolysis temperatures (300 and 600 °C) on the biochemical properties of two mining soils in the Portman area (Spain) in the presence or absence of vegetation. Soils were amended with the RM, the two biochars and a mixture of the rabbit manure and biochars (50/50 w/w) at a rate of 10% in a mesocosms experiment to study the changes in dehydrogenase, phosphomonoesterase, β-glucosidase activities, geometric mean of enzyme activities (GMea) and soil microbial biomass (SMB). Changes in individual enzyme activities were not always consistent. However, when using the GMea as a measure of soil quality, our results showed an increase in the GMea (217–360 times) after the addition of rabbit manure to mining soils, while this increase was from 81–270 times following the addition of rabbit manure with biochar prepared at 300 °C. Therefore, the use of biochar prepared at low temperatures could be a promising direction for the improvement of soil quality and soil carbon sequestration.