Addition of soluble organic substrates to soil has been shown to either increase or restrict the rate of microbial CO2–C evolution. This has been attributed to a priming effect resulting from accelerated or decreased turnover of the soil organic matter including the soil microflora. We investigated microbial responses to small glucose-C additions (10–50 μg C g1 soil) in arable soils either amended or not with cellulose. An immediate CO2–C release between 0 and 69 h (equivalent to 59% of glucose-C applied) was measured. However, only half of the CO2–C respired could be attributed to the utilisation of glucose-C substrate, based on the percentage of 14C–CO2 evolved after the addition of a 14C-labelled glucose tracer. Thus, although no evidence of an immediate release of ‘extra’ C above the rate applied as glucose-C was observed, the pattern of decomposition for 14C-glucose suggested utilisation of an alternate C source. Based on this, a positive priming effect (1.5 to 4.3 times the amount CO2–C evolved that was attributed to glucose-C decomposition) was observed for at least 170 h in non-cellulose-amended soil and 612 h in cellulose-amended soil. Two further phases of microbial activity in cellulose-amended soils were attributed to either activation of different microbial populations or end-product inhibition of cellulase activity after glucose addition. During these subsequent phases, a negative priming effect of between 0.1 and 1.5 times was observed. Findings indicate that the response of the microbial community to small additions of soluble organic C substrate is not consistent and support the premise that microbial response varies in a yet to be predicted manner between soil type and ecosystems. We hypothesise that this is due to differences in the microbial community structure activated by the addition of organic C and the timing of soluble organic substrate addition with respect to the current dissolved organic C status of the soil.