A watershed analysis of nonpoint-source pollution associated with sugarcane (Saccharum officinarum L.) production was conducted. Runoff water samples following major rainfall events from two representative sugarcane fields (SC1 and SC2) were collected and analyzed. The impact of runoff on two receiving water bodies, St. James canal (SJC) and Bayou Chevreuil (BC) in a drainage basin (Baratarian Basin), was studied. Results show that runoff flow/rainfall ratios at the SC1 were significantly higher (P < 0.0001, n = 14) than at the SC2, probably mainly due to higher sand content and higher infiltration rate of surface soil at the SC2. In runoff water samples, total suspended solids (TSS) showed a significant correlation with the concentrations of N and P. Sugarcane runoff showed a direct impact on the SJC and BC locations where seasonal variations of pollutant concentrations in the waters followed the patterns of runoff loadings. Swamp forest runoff (SFR) location showed a buffering effect of forested wetlands on water quality with the lowest measured pollutant concentrations. The ratios in total N/total P and in inorganic N/organic N in runoff waters indicated that fertilization in spring greatly contributed to the temporal increase of N loadings, especially in forms of inorganic N. Isotope signature of (15)N-nitrate in the water samples verified that the nitrate was derived from fertilizers and was consumed during transportation. Both N and P concentrations in the receiving water bodies were above the eutrophic level. During the study period, herbicide concentrations in the receiving water bodies rarely exceeded the drinking water standards.