Buoyancy control and air breathing in royal knifefish (Chitala blanci) and a new hypothesis for the early evolution of vertebrate air-breathing behaviors.

We present the first description of inspiration-first air breaths in royal knifefish, Chitala blanci, a ray-finned fish known to use four-stroke air breaths. Four-stroke breaths are used by nearly all ray-finned fish species that use their gas bladder to breathe air and are the ancestral breath type of ray-finned fishes. Interestingly, one such species, Amia calva, is known to perform two distinct breath types. Amia use four-stroke breaths when they need more oxygen and performs inspiration-first breaths to restore buoyancy. We observed that C. blanci also performs inspiration-first breaths and tested whether the two breath types are performed for the same functions in C. blanci as they are in Amia. We recorded the frequency of each breath type when exposed to aquatic hypoxia and two conditions of oxygen availability. We found that C. blanci performed more four-stroke breaths (81% ± 15% of total breaths) than inspiration-first breaths when exposed to aerial normoxia but performed more inspiration-first breaths (72% ± 40%) than four-stroke breaths when exposed to aerial hyperoxia. These patterns match those described for Amia and indicate that C. blanci performs four-stroke breaths in response to oxygen depletion and performs inspiration-first breaths to maintain buoyancy. Few studies have examined the role of air-breathing in buoyancy regulation. Decreasing buoyancy, rather than oxygen availability, to stimulate air breaths may reveal that inspiration-first breaths are more common among fishes than we are aware. We consider this possibility and present a new hypothesis for the origin and early evolution of air breathing in vertebrates.