<jats:p>Squid (Lolliguncula brevis) were exercised in a tunnel respirometer during a stepwise increase in water velocity in order to evaluate the anaerobic threshold, i.e. the critical swimming speed above which anaerobic metabolism contributes to energy production. The average anaerobic threshold was found at speeds of 1.5-2 mantle lengths s-1. Above this velocity, alpha-glycerophosphate, succinate and octopine started to accumulate in the mantle tissue. ATP levels fell and phospho-L-arginine was progressively depleted, while the levels of glucose 6-phosphate and inorganic phosphate rose. The finding of a simultaneous onset of anaerobic metabolism in the cytosol and the mitochondria indicates that a limited oxygen supply to the mitochondria elicits anaerobic energy production. This finding is opposite to the situation found in many other vertebrate and invertebrate species, in which energy requirements in excess of aerobic energy production are covered by anaerobic metabolism, with mitochondria remaining aerobic. In L. brevis, swimming at higher speeds is associated with a small factorial increase in metabolic rate based on a high resting rate of oxygen consumption. Pressure recordings in the mantle cavity support this finding, indicating a high basal level of spontaneous activity at rest and a small rise in mean pressure at higher swimming velocity. Bursts of higher pressures from the jet support elevated swimming speeds and may explain the early transition to anaerobic energy production which occurs when pressure amplitudes exceed 1.2-1.5 kPa or when mean pressure rises above 0.22-0.25 kPa. The finding of mitochondrial hypoxia at a low critical speed in these squid is interpreted to be related to their life in shallow coastal and bay waters, which limits the necessity to maintain high swimming velocities. At increased swimming velocities, the animals oscillate between periods of high and low muscular activity. This behaviour is interpreted to reduce transport cost and to permit a longer-term net use of anaerobic resources when speed exceeds the critical value or when the squid dive into hypoxic waters. The simultaneous onset of anaerobic metabolism in the cytosol and the mitochondria emphasizes that squid generally make maximal use of available oxygen under resting conditions, when their energy requirements are the highest among marine invertebrates.</jats:p>