Fluctuations in temperature and resource availability are conditions many organisms contend with in nature. Specific dietary nutrients such as fatty acids play an essential role in reproduction, cold adaptation, and metabolism in a variety of organisms. The present study characterizes how temperature and diet interact to modulate Drosophila physiology and life span. Flies were raised on media containing specific saturated, monounsaturated, or polyunsaturated fatty acids supplements at low concentrations and were placed in varied thermal environments. We found that dietary long-chain polyunsaturated fatty acids improve chill coma recovery and modulate mitochondrial function. Additionally, monounsaturated and polyunsaturated fatty acid food supplements were detrimental to life span regardless of temperature, and antioxidants were able to partially rescue this effect. This study provides insight into environmental modulation of Drosophila physiology and life span.