ESM 2 figure DJL and PGDM.pdf from Oxygen extraction efficiency of the tidally-ventilated rectal gills of dragonfly nymphs

Dragonfly nymphs breathe water using tidal ventilation, a highly unusual strategy in water-breathing animals due to the high viscosity, density and low oxygen (O₂) concentration of water. This study examines how well these insects extract O₂ from the surrounding water during progressive hypoxia. Nymphs were attached to a custom-designed respiro-spirometer to simultaneously measure tidal volume, ventilation frequency and metabolic rate. Oxygen extraction efficiencies (OEE) were calculated across four PO₂ treatments, from normoxia to severe hypoxia. While there was no significant change in tidal volume, ventilation frequency increased significantly from 9.4 ± 1.2 BPM at 21.3 kPa to 35.6 ± 2.9 BPM at 5.3 kPa. Metabolic rate increased significantly from 1.4 ± 0.3 µl O₂ min⁻¹ at 21.3 kPa to 2.1 ± 0.4 µl O₂ min⁻¹ at 16.0 kPa, but then returned to normoxic levels as O₂ levels declined further. OEE of nymphs was 40.1 ± 6.1% at 21.3 kPa, and did not change significantly during hypoxia. Comparison to literature shows that nymphs maintain their OEE during hypoxia unlike other aquatic tidal breathers and some unidirectional breathers. This result, and numerical models simulating experimental conditions, indicate that nymphs maintain these extraction efficiencies by increasing gill conductance and /or lowering internal PO₂ to maintain a sufficient diffusion gradient across their respiratory surface.