The racing industry is supported by a predominance of female stablehands and work riders, but few become professional jockeys. Female jockeys have recently had notable race success. No study has assessed whether the sex of the rider may subtly influence racehorse physiology to affect performance. Here, using a validated exercise tracking system (the ‘Equimetre’™) that records many physiological parameters simultaneously, this study characterised racehorse cardiovascular (heart rate, heart rate recovery) and biomechanical (stride length and frequency) parameters at various exercise intensities (slow canter to hard gallop) to address the question whether any parameter varied according to sex of the rider. A total of 530 Thoroughbreds, varying in age (2–7 years old) and sex (including geldings), from one racing yard in Australia, completed a total of 3,568 exercise sessions, monitored by a single trainer, on varying track surfaces (sand, turf, or fibre). Different work riders,103 in total (male, n = 66; female, n = 37) of which n = 43 were current or past registered professional jockeys, participated in the study. Data were analysed using analysis of variation (ANOVA) or mixed-effect models, as appropriate. Sex of the rider did not influence (P > 0.05) racehorse speed nor stride length at any training intensity. Racehorse heart rate and peak heart rate increased with training intensity (P < .001), with no difference according to sex of rider (P > 0.05). Racehorse heart rate recovery was influenced by sex of the rider, but only at the extremes of the reversed, usual training intensity on each surface (e.g. heart rate after galloping on sand was significantly lower with male riders, P = 0.03). Finally, analysis of 52,464 race results indicated a similar chance of a top-three placing for male and female jockeys. In conclusion, this study, using objectively obtained data, demonstrates for the first time no overt effect of the rider’s sex on racehorse physiology in training and performance in racing. Such data could encourage greater female participation in racing and improve access of female jockeys to better quality mounts in racing events.