Here it is evaluated the relationship between substrate-induced respiration (SIR) and microbial biomass C (C mic , estimated by chloroform fumigation–extraction) in order to establish SIR as a quick technique to determine microbial biomass in vermicomposting processes. For this, there were designed continuous feeding reactors in which new layers of manure were added sequentially to form an age gradient inside the reactors. Six reactors were set up with and without earthworms (three reactors per treatment). In reactors, with and without earthworms, values of C mic and SIR ranged from 1690 to 42,900μgg −1 dw and from 43 to 2300μg CO 2 h −1 g −1 dw, respectively. SIR was significantly related to C mic (r=0.63, P<0.0001). It is proposed an equation to convert SIR values into C mic , SIR (μg CO 2 h −1 g −1 dw)=25.97+0.04 C mic (μgg −1 dw) which will be useful for comparison between studies which used different techniques for determining microbial biomass.