This study examined the ability of nearinfrared spectroscopy (NIRS) to noninvasively determine changes to muscle oxygenation in the resting horse. Five horses had (NIRS) performed over extremity muscle while under general anaesthesia, first with 8 min limb ischaemia, then systemic hypoxaemia for 5 min. A second group of 6 awake horses had NIRS performed over extremity muscle while being administered hypoxic gas (F1O2 0.10) for 5 min, and after return to steady state, limb ischaemia was induced for an additional 5 min. In the anaesthetised horses' ischaemia induced marked and significant muscle deoxygenation of haemoglobin/myoglobin (P<0.01), with corresponding arterial saturation decreasing from 98.9 to 81.9%. Hypoxaemia induced small yet significant muscle deoxygenation (P<0.01) that was 3.2% of the ischaemia deoxygenation signal, with a corresponding decrease in arterial saturation from 98.6 to 90.4%. In the awake horses muscle deoxygenation was not detectable during hypoxia despite reduction of arterial saturation from 97.8 to 86.8%, whereas ischaemia induced rapid and significant deoxygenation of muscle (P<0.05), with corresponding reduction of venous saturation from 78.4 to 75.4%. In neither group of horses was there evidence of cytochrome aa3 reduction, despite complete ischaemia forup to 8 min. NIRS changes in the resting horse muscle clearly differed between ischaemia and hypoxaemia, and can readily show muscle deoxygenation in clinically relevant hypoxaemia in the horse under anaesthesia. Further, as the deoxygenation signal induced by ischaemia was clearly detectable above a background movement artefact, NIRS application to study of muscle oxygenation in the working horse should be explored.