The Gascogne region (SW of France) is cultivated for more than 75% of the area. 83 samples of stream bed sediments were collected in three main Gascogne river basins (Gers, Save and Touch, left tributaries of the Garonne river) to evaluate the impact of agricultural practices on trace elements behavior. Eight potential harmful elements (PHE) (Cr, Co, Ni, Cu, Zn, As, Cd and Pb), four reference elements for normalization (Sc, Cs, Al and Fe) and four major elements (Mn, Ca, Mg and P) were considered. The average trace element concentrations in the fine fractions (<63 microm) are in the decreasing order: Zn>Cr>Ni>Pb>Cu>Co>As>Sc>Cs>Cd. Geochemical investigations and an original approach combining regression analysis and chemical sequential extraction allowed to select the most adequate reference material (regional molasse) and reference element (Cs) for normalization procedure. The enrichment factor (EF) is generally lower than 2.5, particularly for Cr, Ni, Cu, As, Zn; however, 23% of the sampling stations are more contaminated (2.5<EF<4.5), particularly for Cd, Pb and Co. The PHE in the Gascogne river sediments are mainly originated from natural weathering processes; nevertheless, anthropogenic contribution could represent up to 34% of the total sediment content. For lead, geochemical and isotopic methods gave very similar anthropogenic contributions (24% and 22%, respectively). The enrichment of Cu, Pb, Zn, Co, As, Ni, Cr was mainly related to global and local atmospheric deposition of industrial emissions and gasoline combustion, and was associated to forested catchments. All PHE's are controlled by clay and oxi-hydroxides minerals. Cd was the only PHE enriched downstream cultivated catchments and this enrichment was linked to Ca and P. This indicates a major origin of Cd from fertilizer inputs and a main control by carbonate minerals.