It is widely recognized that the efficiency of substrate C use in acid and/or disturbed soils by soil microorganisms is relatively low based on the observation that metabolic quotients (qCO2) are usually high in these soils. In the present study, threshold concentrations of glucose, at which the ratio of respiration to assimilation by soil microorganisms began to increase, were comparatively analyzed using two soils differing in pH and disturbance, a Japanese arable soil disturbed by cultivation and a strongly acidic Japanese forest soil. Varying concentrations of glucose C, generally less than those in the microbial biomass C, were added to the two soils. The ratio of respired to utilized glucose C remained at approximately 20% when lower concentrations of glucose were added and respiration rates did not increase (zero-order types), whereas the ratio increased when the concentrations of added glucose exceeded a certain level and respiration rates increased (growth-associated types). The substrate-induced respiration rate a few hours after the addition of glucose increased only for the growth-associated types, although chloroform-labile C increased in both types as the concentrations of added glucose increased. The results clearly confirmed the presence of a threshold concentration of glucose above which the ratio of respiration to assimilation increased. The threshold concentrations in Japanese arable and forest soils were lower than the concentration previously reported in a moderately acidic Kazakh forest soil. The lower threshold concentrations observed in the Japanese arable and forest soils are considered to result from different microbial growth characteristics after the addition of glucose linked with a shorter lag period before the exponential increase of the respiration rate and a lower ratio of substrate induced respiration rate to biomass C, respectively. The results suggest that the efficiency of substrate C use in acid and disturbed soils is relatively low in situations where higher concentrations of substrates are occasionally supplied with temporal C ‘flushes’, such as may occur in the rhizosphere or in the vicinity of plant residues.