Abstract   Ecotoxicological studies mainly consist of single-species experiments evaluating the effects of a single stressor. However, under natural conditions aquatic communities are exposed to a mixture of stressors. The present study aimed to identify how the toxicity of zinc (Zn) is affected by increased temperature and increased phosphorus (P) supply and how these interactions vary among species, functional groups, and community structure and function. Aquatic microcosms were subjected to 3 Zn concentrations (background, no Zn added, and 75 and 300 μg Zn/L), 2 temperatures (16–19 and 21–24 °C), and 2 different P additions (low, 0.02, and high, 0.4 mg P L−1 wk−1) for 5 wk using a full factorial design. During the study, consistent interactions between Zn and temperature were only rarely found at the species level (4%), but were frequently found at the functional group level (36%), for community structure (100%) and for community function (100%; such as dissolved organic carbon concentrations and total chlorophyll). The majority of the Zn × temperature interactions were observed at 300 μg Zn/L and generally indicated a smaller effect of Zn at higher temperature. Furthermore, no clear indication was found that high P addition by itself significantly affected the overall effects of Zn on the community at any level of organization. Interestingly, though, 90% of all the Zn × temperature interactions observed at the species, group, and community composition level were found under high P addition. Collectively, the results of our study with the model chemical Zn suggest that temperature and phosphorus loading to freshwater systems should be accounted for in risk assessment, because these factors may modify the effects of chemicals on the structure and functioning of aquatic communities, especially at higher levels of biological organization. Environ Toxicol Chem 2018;37:2413–2427. © 2018 SETAC