Our objective was to find the relationship between benthic index of biotic integrity (B-IBI) and land use in the Xitiaoxi stream catchment, Zhejiang Province, China. Benthic macroinvertebrae assemblages were collected from 64 sites in May, 2004 and 2005. The B-IBI was composed of seven metrics: total taxa, EPT taxa, Coleoptera%, three dominant taxa, (Hydropsychidae/Trichoptera) %, filterers% and biotic index. Land use upstream of each site at three spatial scales was characterized by analysis of a 2003 satellite image. Results showed that the percentage of the broad-leaved forest had significantly positive correlation with B-IBI at sub-basin (r= 0.333,p<0.001),riparian(r=0.589,p<0.001)and local(r=0.618,p<0.001)scale. While the percentage of the cropland had the strongest negative correlation with B-IBI at sub-basin (r=-0.527, p<0.001), riparian (r=-0.665, p<0.001) and local (r=-0.696, p<0.001) scale. B-IBI declined linearly as the increasing of the percentage of the cropland but stopped at thresholds of 27% at sub-basin scale, 57% at riparian scale, and 59% at local scale. Our results indicated that the selection of a correct spatial scale is important to explain the relationship between biological condition of Xitiaoxi stream and cropland use.