The proximity of freshwater ecosystems to anthropogenic activities makes them one of the most threatened environments by plastic pollution in the form of microplastics (MPs). Therefore, it is crucial to identify the primary drivers of MP dynamics in rivers to enhance their management. This work analyzed the concentration of MPs in water and sediments and evaluated the influence of land use and its relationship with the main biotic indices employed to assess the water quality of rivers. This research was carried out in four different catchments, with three sampling points established in each river basin. The results revealed that MPs were ubiquitous across all locations, with concentrations ranging from 0.10 to 35.22 items m⁻³ in waters and from 26 to 643 items Kg⁻¹ in sediments. The highest concentration of MPs both in water and sediments were found in the Lagares River (35.22 items m⁻³ and 643 items Kg⁻¹), while the lowest concentrations were found in the Miñor River for water (0.10 items m⁻³) and Tea River for sediments (138 items Kg⁻¹). Urbanization degree was identified as the primary driver of MP pollution in water, whereas population density correlated with sediment pollution levels. These findings explain the elevated MPs abundance in the more urbanized and populated Gafos and Lagares rivers compared to the relatively pristine Miñor and Tea rivers. Furthermore, the presence of MPs in sediments was found to negatively impact the most sensitive benthic macroinvertebrate taxa, as evidenced by lower values of the IASPT and EPT indices at sampling points with higher sediment MPs concentrations (Gafos and Lagares).