Urbanization alters the hydrological and ecological dynamics of watercourses, compromising habitat integrity, water quality, and associated ecosystem services. In this context, this study aimed to assess the environmental integrity of urban streams in Tangará da Serra, Mato Grosso (MT), by relating morphometric characteristics, land use and cover, habitat ecological quality, and physicochemical water parameters to inform environmental management and planning strategies. Six urban sub-basins within the Sepotuba River basin were evaluated, covering headwater, mid-reach, and mouth sections. Ecological quality was determined using the Rapid Habitat Diversity Assessment (ARDH) protocol, complemented by morphometric characterization, spatial analysis of land use and cover, and assessment of physicochemical water parameters. Relationships between environmental variables were investigated using Spearman’s correlation and Principal Component Analysis (PCA). The results revealed a gradient of human alteration associated with urbanization, with ARDH scores ranging from 0 to 87. Forest cover showed a positive correlation with ecological integrity (ρ = 0.580; p < 0.05), whereas urban area showed a negative correlation (ρ = -0.569; p < 0.05). PCA summarized 60.2% of the environmental variability within the first two components, highlighting the contrast between urbanization and ecological integrity. São Pedro Stream exhibited the most critical condition, with an ARDH score of zero and the absence of permanent flow, while the Estaca and Figueira streams presented conditions warranting priority conservation. Based on this diagnosis, specific technical guidelines were established regarding environmental restoration, sustainable land-use guidance, land-use planning, enforcement, monitoring, and environmental education. It is concluded that the integration of scientific diagnosis and technical application constitutes an appropriate strategy for identifying priority areas and supporting management actions for the urban streams of Tangará da Serra, MT.