The aims of this study were (1) to identify which physical performance tests could best explain the development of fatigue during a simulated ambulance work task, (2) to investigate the effect of height and weight and (3) to investigate in what respects these findings differ between female and male ambulance personnel. Forty-eight male and 17 female ambulance personnel completed a test battery assessing cardio-respiratory capacity, muscular strength and endurance, and co-ordination. The subjects also completed a simulated ambulance work task -- carrying a loaded stretcher. The work task was evaluated by development of fatigue. Univariate and multiple regression analyses were used to investigate to what extent the tests in the test battery were able to explain the variance of developed fatigue. The explained variance was higher for female than for male ambulance personnel (time > 70% of HRpeak: R2 = 0.75 vs 0.10, accumulated lactate: R2 = 0.62 vs 0.42, perceived exertion: R2 = 0.75 vs 0.10). Significant predictors in the models were VO2max, isometric back endurance, one-leg rising, isokinetic knee flexion and shoulder extension strength. Height, but not weight, could further explain the variance. The high physical strain during carrying the loaded stretcher implies the importance of investigating whether improved performance, matching the occupational demands, could decrease the development of fatigue during strenuous tasks.