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1.4.2 Respirometry
Once at the field station, birds were maintained in separate cages (39 cm x 43 cm x 31 cm) supplied with food (sunflower seed) and water ad libitum until measurements were madę. Cages were kept in a room receiving natural light through a window and maintained quiet to avoid disturbance. At around 13:00, we began Msum trials by measuring two birds in parallel using FoxBox oxygen analyzers (Sabie Systems, Las Vegas, NV, USA). This was followed by a second trial on the remaining two birds, which began before 15:00. Briefly, birds were first weighed (± 0.1 g) and body temperaturę was measured with a thermocouple reader (Omega model HH-25KC, NIST-traceable, Omega, Montreal, Qc, Canada) using a copper-constantan thermocouple inserted into the cloacae approximately 10 mm deep. Then, birds were put in a stainless Steel metabolic chamber fitted with a perch and were exposed to helox gas (21% oxygen, 79% helium) using an average flow ratę of 1109 ml.min*1 controlled by mass flow valves (Sierra Instruments, Side-Trak® Model 840, Monterey, CA, USA). We recorded oxygen consumption of each bird using a sliding cold exposure protocol (Swanson et al., 1996) with a decrease in ambient temperaturę of 3°C every 20 minutes, starting at 0°C in winter and at 6°C in summer. We ended the trials when birds became hypothermic, which was easily identifiable in real time as a steady decline in oxygen consumption for several minutes. Body temperaturę was measured again immediately after taking birds out of their chambers. We assumed a bird had reached its Msum when body temperaturę after a trial was < 38°C (Cooper & Gessaman, 2005) (mean body temperaturę after Msum measurement = 33.6 ± 0.2°C). Data from individuals showing a body temperaturę above this threshold were discarded. Birds were weighed again after measurements and the average body mass was used for the Msum analysis. Birds were then brought back to their cage with food and water ad libitum until BMR measurement starting at 19:00.
Each day, all four birds had their BMR measured simultaneously ovemight (from 19:00 to 06:00). Individuals were maintained at 30°C throughout the trial (within the thermoneutral zonę for this species, Rising & Hudson, 1974) and received a constant flow of air. Birds were weighed before and after measurements and average mass was used in BMR analyses.
Oxygen analyzers were adjusted each day to 20.95% using C02-free diy air and mass flow valves were carefully calibrated for air and helox using a bubble-O-meter (Dublin, OH, USA)
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