Birds naturally have high circulating blood glucose concentrations compared to other vertebrates. Several mechanisms have been proposed to explain their high levels including the lack of an insulin responsive glucose transport protein, higher circulating glucagon concentrations, as well as a reliance on lipid oxidation to fuel the high metabolic demands for flight. We suspected the latter may result in the production of the gluconeogenic precursor, glycerol. Therefore, we examined the hypothesis that gluconeogenesis, via glycerol, contributes to the naturally high glucose concentrations in birds (Madiraju et al., 2014). We captured seven mourning doves, Zenaida macroura, in Tempe, AZ, USA and acclimated the birds to captivity for two weeks. In this crossover design study, doves received either an oral inhibitor of gluconeogenesis (150 or 300 mg/kg metformin) or water (50 ul) each week. We measured blood glucose concentrations using a glucometer at baseline, 30, 60 and 120 minutes following the oral dose. In contrast to mammals and chickens, 300 mg/kg metformin did not alter blood glucose (p>0.05) and 150 mg/kg metformin significantly increased blood glucose concentrations (p=0.043) compared to the oral bolus of water. To examine whether the low dose of metformin stimulated glycogenolysis, thus causing the hyperglycemic effect, we administered the low dose of metformin along with an inhibitor of glycogenolysis, 2.5 mg/kg 1,4-dideoxy-1,4-imino-D-arabinitol (DAB), which prevented the hyperglycemic response (p>0.05 vs. water). These data suggest that low doses of metformin activate glycogenolysis. It is possible that glycogenolysis is also activated at the higher dose, but glycogen may be depleted early on resulting in no measurable changes, given the present study design. In conclusion, and in contrast to the hypothesis, mourning doves may not rely on gluconeogenesis to maintain their naturally high blood glucose concentrations under fed conditions, although further studies with more specific gluconeogenic antagonists and under fasted conditions may be needed to confirm these findings.