The diversity, structure, and function of bacterial communities in freshwater rivers can give insight into the health status of those systems. This project aimed to conduct next-generation sequencing of water and sediments from the upper Hawkesbury River (NSW, Australia); assign taxonomies and infer functional profiles from the metagenomic information, and consider the utility of this work in assessing whether a freshwater system is healthy. To achieve this, we conducted routine sampling over twelve months. 16S rRNA extracted from the samples were sequenced on an Illumina MiSeq platform. Sequences were quality filtered and paired with the DADA2 workflow in R. Taxonomies were assigned with the SILVA v138 database, and functional profiles were predicted using the Piphillin server to infer KEGG pathways. This research generated a comprehensive bacterial community profile for the upper Hawkesbury River. It uncovered the presence of bacterial taxa that are important ecologically, agriculturally, and socially. The functional profiling predicted a broad range of ecological functions, including a suggestion of poor human disease outcomes in the local area. The findings suggest that metagenomic methods, as used in this project, are useful in generating a “snapshot” of the microbial functioning of a freshwater system.