Keeling plots for hummingbirds:: a method to estimate carbon isotope ratios of respired CO2 in small vertebrates

The carbon isotope composition of an animal's breath reveals the composition of the nutrients that it catabolizes for energy. Here we describe the use of Keeling plots, a method widely applied in ecosystem ecology, to measure the delta(13)C of respired CO2 of small vertebrates. We measured the delta(13)C of Rufous Hummingbirds (Selasphorus rufus) in the laboratory and of Mourning (Zenaida macroura) and White-winged (Z. asiatica) Doves in the field. In the laboratory, when hummingbirds were fed a sucrose based C3 diet, the delta(13)C of respired CO2 was not significantly different from that of their diet (delta(13)C(C3 diet)). The delta(13)C of respired CO2 for C3 fasted birds was slightly, albeit significantly, depleted in delta(13)C relative to delta(13)C(C3 diet). Six hours after birds were shifted to a sucrose based C4 diet, the isotopic composition of their breath revealed that birds were catabolizing a mixture of nutrients derived from both the C3 and the C4 diet. In the field, the delta(13)C of respired CO2 from Mourning and White-winged Doves reflected that of their diets: the CAM saguaro cactus (Carnegeia gigantea) and C3 seeds, respectively. Keeling plots are an easy, effective and inexpensive method to measure delta(13)C of respired CO2 in the lab and the field.