Abstract: Using a microplate fluorimetric assay method, five fertilization treatments, i.e. nofertilizer control (CK), sole application of nitrogen (N), balanced application of nitrogen, phosphorus, and potassium fertilizer (NPK), application of pig manure (M), and combination of pig manure with balanced chemical fertilizer (MNPK) were selected to investigate the effects of different long-term fertilization regimes on the activity of five enzymes (β-1,4-glucosidase, βG; cellobiohydrolase, CBH; β-1,4-xylosidase, βX; β-1,4-Nacetylglucosaminidase, NAG; acid phosphatase, AP) in a red soil sampled from Qiyang, Hunnan Province. The results showed that compared with CK treatment, N treatment had no impact on βG, βX, CBH, and NAG activities but reduced AP activity, while NPK, M and MNPK treatments increased the activities of all the five enzymes. Correlation analysis indicated that all the five enzyme activities were positively correlated with the content of nitrate (r=0.465-0.733), the content of available phosphorus (r=0.612-0.947), soil respiration (r=0.781-0.949) and crop yield (r=0.735-0.960), while βG, CBH and AP were positively correlated with pH (r=0.707-0.809), only AP was significantly correlated with dissolvable organic carbon (r=-0.480). These results suggested that the activities of the measured enzymes could be used as indicators of red soil fertility under different fertilization regimes, but the five enzymes tested provided limited information on the degree of acidification induced by application of mineral nitrogen.