The distribution and traits of macroinvertebrates provide important information about stream water and habitat quality that can support conservation of threatened and endangered species of fish and mussels. Our study uses macroinvertebrate data collected along a longitudinal gradient and over a five-year period in Holly Creek, a major tributary of the biodiverse Conasauga basin (GA, USA), in which several rare fish and mussel species are the restoration focus. These rare taxa of interest are inherently difficult to sample and thus require proxy-indicators. Macroinvertebrates will serve as that proxy-indicator because of how much information we can extract from them using our data and outside databases. We have collected macroinvertebrates and abiotic data (pebble count, canopy cover, and water chemistry) for five years at six sites in Holly Creek. We are using macroinvertebrate functional traits extracted from the CONUS database in addition to biodiversity indices to determine the potential longitudinal patterns and temporal changes in Holly Creek’s water quality and riffle habitat. With this information, we can begin to characterize the aquatic conditions for species of conservation concern. The distribution and abundance of macroinvertebrate traits can be used to infer which basal resources and physical/chemical drivers (e.g., sedimentation) are present at a site and over time. We will use ordination analyses to determine upstream to downstream patterns and test whether downstream locations are more impacted by land use change through time. We expect that these data will help establish the baseline habitat conditions for the species of interest and will guide conservation of rare species in other systems. Ultimately, our project will use a suite of ecological responses to evaluate if restoration actions are having a measurable impact on Holly Creek and aim to provide insights into specific restoration strategies.