The effect of pesticide napropamide (N,N-diethyl-2-(1-naphthalenyloxy) propanamide) on soil microorganisms for long-term (56 d) was assessed by monitoring changes in soil microbial biological responses. Soils were treated with napropamide at 0, 2, 10, 20, 40 and 80 mg/kg soil and sampled at intervals of 1, 3, 7, 14, 28, 42 and 56 d. The average microbial biomass C declined in napropamide-treated soils as compared to the control. The same trend was observed on microbial biomass N after napropamide application. We also determined the basal soil respiration (BSR) and observed a high level in soils treated with napropamide during the first 7 d of experiment. But with the passage of incubation time, BSR with napropamide decreased relative to the control. Application of napropamide at 2-80 mg/kg soil had inhibitory effects on the activity of urease and invertase. Activity of catalase was enhanced during the initial 7 d of napropamide application, but soon recovered to the basal level. The depressed enzyme activities might be due to the toxicity of napropamide to the soil microbial populations. To further understand the effect of napropamide on microbial communities, a PCR-DGGE-based experiment and cluster analysis of 16S rDNA community profiles were performed. Our analysis revealed an apparent difference in bacterial-community composition between the napropamide treatments and control. Addition of napropamide apparently increased the number of bands during the 7-14 d of incubation. These results imply that napropamide-induced toxicity was responsible for the disturbance of the microbial populations in soil.