Despite increasing human activity and risk of fuel spills in Antarctica, little is known about the impact of fuel on Antarctic marine fauna. The authors performed both single- and multi-species (whole community) acute toxicity tests to assess the sensitivity of an Antarctic coastal zooplankton community to the water-accommodated fraction of Special Antarctic Blend diesel. Single-species tests using abundant copepods Oncaea curvata, Oithona similis, and Stephos longipes allowed comparisons of sensitivity of key taxa and of sensitivity estimates obtained from traditional single-species and more novel multi-species tests. Special Antarctic Blend diesel caused significant mortality and species compositional change in the zooplankton community within 4 d to 7 d. The sensitivity of the community also increased across the summer sampling period, with decreasing 7-d median lethal concentration (LC50) values for total petroleum hydrocarbons (TPH): 1091 µg TPH/L in early January 2011, 353 µg TPH/L in mid January 2011, and 186 µg TPH/L in early February 2011. Copepods showed similar sensitivities to Special Antarctic Blend diesel in single-species tests (7-d LC50s: O. curvata, 158 µg TPH/L; O. similis, 176 µg TPH/L; S. longipes, 188 µg TPH/L). The combined use of single- and multi-species toxicity tests is a holistic approach to assessing the sensitivity of key species and the interactions and interdependence between species, enabling a broader understanding of the effects of fuel exposure on the whole zooplankton community. Environ Toxicol Chem 2014;9999:1-9. © 2014 SETAC.