Sub-Antarctic islands have been subjected to petroleum hydrocarbon spills, yet no information is available regarding the toxicity of petroleum hydrocarbons to these subpolar soils. The purpose of the present study was to identify soil biogeochemical toxicity end points for petroleum hydrocarbon contamination in sub-Antarctic soil. Soil from Macquarie Island, a sub-Antarctic island south of Australia, was collected and exposed to 10 concentrations of Special Antarctic Blend (SAB) diesel fuel, ranging from 0 to 50,000 mg fuel/kg soil, for a 21-d period. The sensitivity of nitrification, denitrification, carbohydrate utilization, and total soil respiration to SAB fuel was assessed. Potential nitrification activity was the most sensitive indicator of SAB contamination assessed for nitrogen cycling, with an IC20 (concentration that results in a 20% change from the control response) of 190 mg fuel/kg soil. Potential denitrification activity was not as sensitive to SAB contamination, with an IC20 of 950 mg fuel/kg soil for nitrous oxide production. Nitrous oxide consumption was unaffected by SAB contamination. Carbohydrate utilization (respiration caused by sucrose) was a more sensitive indicator (IC20, 16 mg fuel/kg soil) of SAB contamination than total respiration (IC20, 220 mg fuel/kg soil). However, total soil respiration was a more responsive measurement end point, increasing soil respiration over a 72-h period by 17 mg of CO2, compared to a change of only 2.1 mg of CO2 for carbohydrate utilization. Our results indicate that IC20s varied between 16 to 950 mg fuel/kg soil for Macquarie Island soil spiked with SAB diesel fuel. These results indicate that current cleanup levels derived from temperate zones may be too liberal for soil contamination in sub-Antarctic islands.