Magnitude of the Root effect in red blood cells and haemoglobin solutions of fishes: a tribute to August Krogh

Aim: The ability of high carbon dioxide tensions or low pH to reduce blood oxygen binding even at high oxygen tensions, first observed by August Krogh and Isabella Leitch in 1919 and now known as the Root effect, was studied in red blood cells and haemoglobin solutions of several fish species. Methods: Red blood cells in physiological saline were acidified at atmospheric oxygen tension by increasing carbon dioxide tensions and the percentage decrease in oxygen content was used to quantify the Root effect. Haemoglobin was incubated in air-equilibrated citrate buffers between pH 5 and 7 and the percentage decrease in oxygen saturation relative to pH 8 determined by spectral deconvolution. Results: The maximal magnitude of the Root effect in citrate-buffered haemoglobin solutions closely matched the value in blood or red blood cells of 11 vertebrates over a Root effect range between 3 and 80%. Contrary to previous reports, there was no evidence for a significant Root effect in red blood cells or haemoglobin solutions of the wels catfish, but a significant Root effect under both conditions in the Siberian sturgeon. Conclusions: Under the conditions employed in this study, the maximal Root effect of citrate-buffered haemoglobin solutions closely resembles the maximal Root effect in red blood cells. This strengthens previous studies on the evolution of the Root effect and its role in oxygen concentration at the retina and swimbladder of a large number of fishes that were based on Root effect measurements in haemoglobin solutions.