A long-term exposure outdoor microcosm study was conducted to evaluate the effects of zinc (Zn) on zooplankton, phytoplankton, and periphyton in a freshwater system. Five Zn treatment concentrations (nominal: 8, 20, 40, 80, and 160 μg/L Zn) and an untreated control with 3 replicates each were used. Various physical and chemical characteristics of the microcosms and biological assessment endpoints (e.g., total abundance, group abundance, species richness, chlorophyll a, etc.) were measured to determine the effects of Zn over time. In general, physical and chemical characteristics (e.g., total dissolved solids, total suspended solids, dissolved oxygen, pH, dissolved organic carbon) of water fluctuated over time, but they were not significantly different within treatments and controls during the study. Zinc significantly affected the population dynamics and community structure of plankton. The effects occurred 7 d after initial treatment exposures began and continued to the end of the treatment phase, especially at the high treatment concentrations. Total and group abundance, species richness, the Shannon index, and chlorophyll a concentrations for high Zn treatment concentrations were significantly lower than the controls during the treatment phase. The no-observed-effect, lowest-observed-effect, and median effect concentrations were generally lower than the literature-reported results from single-species toxicity tests for fish and invertebrates, suggesting that plankton are more sensitive to Zn than planktivores. Although primary producers play an important role in the ecosystem, they have not been consistently incorporated into numerical environmental quality criteria for freshwater organisms, at least in the United States. The results of the present study are useful for development of environmental quality guidelines for freshwater ecosystems and ecological risk assessment.