We examined the consequences of deforestation and pasture establishment on H, O and Fe isotope cycling in the hydrosphere, with focus on small watersheds from the central and eastern part of Brazilian Amazonia. The δ18O- δD relationships in stream water samples show that the evaporation effect is greater in pasture than in forest, thereby highlighting the impact of deforestation on atmospheric water circulation and rainfall. A clear effect of deforestation was also found on the dissolved load of these streams. Water temperature, pH, conductivity, major cations and dissolved carbon organic content are lower in forest streams compared to those from deforested areas. Dissolved (i.e., filtrate <0.45 μm) iron concentrations are typically higher in these tropical forest streams. They also show δ57Fe signatures commonly higher than that of the continental crust (i.e., > 0.1‰). In these black waters enriched in organic matter, the heavier iron isotope composition in the forested areas is related to the oxidized Fe strongly bound to colloidal organic matter. On the other hand, dissolved iron from streams draining older deforested areas shows isotopic compositions usually lighter than the continental crust (δ57Fe < 0.1‰). The negative δ57Fe signatures of these areas are likely caused by deforestation involving erosion and rejuvenation of soils in valleys, which leads to the occurrence of a partial iron reduction in the soils. These modifications increased the proportion of isotopically light, dissolved and more reduced Fe in the streams. Those effects are less obvious on more recent pastures since they display more chemical and isotopic scatter along the sampling months. In a stream flowing through an area undergoing slash and burned deforestation within the year, dissolved iron concentrations and isotopic compositions are much more scattered, with δ57Fe values varying by >10 ‰ as a result of fire or heavy rain events. This likely depicts the disruption of the iron cycling in the water-soil-plant system with strong changes in the soil mineralogy, organic matter characteristics and biomass activity resulting from the forest fire. Our results suggest that more than a decade is needed to reach a new dynamic steady state of the Fe biogeochemical cycle in pastures. Hence, the contrasted Fe isotopic compositions between the streams draining pristine tropical forests and those from deforested areas indicate that, like H and O isotopes, Fe isotopes are a sensitive indicator of the transformations affecting the biogeochemical cycling of tropical environment in response to deforestation. However, in contrast to hydrogen and oxygen isotopes that reveal the exchanges between surface waters and atmosphere, iron isotopes rather highlight the physical-chemical reactions occurring between surface waters and the soils, but with the biomass mediation for all three isotopic tracers.