Oxygen binding properties of backswimmer (Notonectidae, Anisops) haemoglobin, determined in vivo

[Display omitted] ► The oxygen equilibrium curve (OEC) of Anisops haemoglobin is determined in vivo using a biotonometric technique. ► The in vivo OEC displays an unusual degree of triphasy and a high level of cooperativity (Hill’s n = 15.2). ► Comparison with an in vitro OEC shows both oxygen equilibrium curves are qualitatively similar. Aquatic backswimmers ( Anisops spp.) collect oxygen from the atmosphere in order to breathe underwater, carrying it within a bubble of air on the ventral surface of their body and bound within haemoglobin-filled cells inside their abdomen. These oxygen stores are interconnected via the abdominal spiracles and the tracheal system. Fibre optic oxygen probes were used to measure PO 2 changes within the air bubbles of submerged backswimmers ( Anisops deanei) and these measurements were transformed into in vivo haemoglobin–oxygen equilibrium curves (OECs) using a biotonometric approach. The haemoglobin displayed a triphasic, highly sigmoid OEC with a P 50 of 3.90 kPa. Comparisons made with a previous in vitro analysis of Anisops haemoglobin demonstrate that while the apparent cooperativity and oxygen affinity are considerably higher in vivo, both measurements share unusual Hb–O 2 binding characteristics. The affinity and cooperativity of the backswimmers’ haemoglobin appears adaptive as it lengthens dives and promotes neutral buoyancy. While there are limitations associated with biotonometry, the in vivo OEC accurately represents the loading and unloading of biologically available oxygen within the backswimmers’ haemoglobin cells. Potential errors associated with determining the OEC are small, as evaluated with sensitivity analyses in numerical models.