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Physical fitness testing is commonplace in schools and in the physical education (PE) curriculum. Despite this, much controversy surrounds the issue of student fitness testing. The Relative Effect of Age (REA) is the potential cognitive and biological variability among individuals of an age- grouped cohort, and its presence within cohorts has previously been identified as affecting an individual’s ability to demonstrate relative proficiency. The aim of this study was to evaluate whether birth-date distributions and gender affect physical education (PE) fitness testing results. 252 (124 male and 128 female) pupils’ PE fitness testing marks were recorded at a large secondary school. 5 groups were formed from the students for that academic year, dividing them into 4 birth quartiles: a) first quartile (January-March); b) second quartile (April-June); c) third quartile (July-September); and d) fourth quartile (October-December). Significant differences were found when the fitness testing marks were compared with the birth-date distribution (F(3, 24) = 5.39; p < .01; ŋ2 = .06; ß = .93) and gender (F(1, 24) = 13.20; p < .01; ŋ2 = .05; ß = .95). The results of the study suggest that there is a difference in fitness testing performance, depending on the birth quartile and gender. Based on the evidence, the paper concludes that much of the fitness testing carried out in PE classes may well represent a disadvantage for students born in the final months of the year. At the same time, gender differences were also found in this type of testing.