AbstractFish community composition is highly dynamic, but generally exhibits loose equilibrium within bounds of community structure in upland stream systems with minimal anthropogenic disturbance. Chronic anthropogenic disturbance can exceed ecological thresholds and shift fish community composition to an alternative state. Therefore, long‐term datasets are critical to understand spatiotemporal dynamics of fish community structure. We quantified temporal changes in fish communities in two Ouachita River systems during watershed alteration (i.e., development and reservoir construction) using a suite of multivariate and univariate analyses to assess changes in fish community structure and taxonomic and functional β‐diversity. We found fish community composition shifted in the Saline River basin possibly in response to human disturbance (e.g., development) and hydrologic variability. Furthermore, functional homogenisation occurred through time. In the Ouachita Headwaters basin, taxonomic homogenisation was evident and two fluvial specialists were not detected during the current period. Taxonomic and functional β‐diversity patterns were not consistent across basins demonstrating the complexity of biotic homogenisation. Our retrospective analyses showed human activities played a role in restructuring fish communities despite minimal invasion of exotic species. Ultimately, our study characterised fish community structure across snapshots of time and demonstrates the importance of increasing the frequency of sampling intervals and continuing long‐term collections with similar methods to enhance the understanding of community dynamics.