Stable isotope analysis of CO₂ in breath indicates metabolic fuel shifts in torpid arctic ground squirrels

Stable carbon isotope ratios (δ¹³C) in breath show promise as an indicator of immediate metabolic fuel utilization in animals because tissue lipids have a lower δ¹³C value than carbohydrates and proteins. Metabolic fuel consumption is often estimated using the respiratory exchange ratio (RER), which has lipid and carbohydrate boundaries, but does not differentiate between protein and mixed fuel catabolism at intermediate values. Because lipids have relatively low δ¹³C values, measurements of stable carbon isotopes in breath may help distinguish between catabolism of protein and mixed fuel that includes lipid. We measured breath δ¹³C and RER concurrently in arctic ground squirrels (Urocitellus parryii) during steady-state torpor at ambient temperatures from − 2 to − 26 °C. As predicted, we found a correlation between RER and breath δ¹³C values; however, the range of RER in this study did not reach intermediate levels to allow further resolution of metabolic substrate use with the addition of breath δ¹³C measurements. These data suggest that breath δ¹³C values are 1.1‰ lower than lipid tissue during pure lipid metabolism. From RER, we determined that arctic ground squirrels rely on nonlipid fuel sources for a significant portion of energy during torpor (up to 37%). The shift toward nonlipid fuel sources may be influenced by adiposity of the animals in addition to thermal challenge.