Differing conclusions have been reached as to how or whether varying heat production has a thermoregulatory function in flying honeybees Apis mellifera. We investigated the effects of air temperature on flight metabolic rate, water loss, wingbeat frequency, body segment temperatures and behavior of honeybees flying in transparent containment outdoors. For periods of voluntary, uninterrupted, self-sustaining flight, metabolic rate was independent of air temperature between 19 and 37 degrees C. Thorax temperatures (T(th)) were very stable, with a slope of thorax temperature on air temperature of 0.18. Evaporative heat loss increased from 51 mW g(-1) at 25 degrees C to 158 mW g(-1) at 37 degrees C and appeared to account for head and abdomen temperature excess falling sharply over the same air temperature range. As air temperature increased from 19 to 37 degrees C, wingbeat frequency showed a slight but significant increase, and metabolic expenditure per wingbeat showed a corresponding slight but significant decrease. Bees spent an average of 52% of the measurement period in flight, with 19 of 78 bees sustaining uninterrupted voluntary flight for periods of >1 min. The fraction of time spent flying declined as air temperature increased. As the fraction of time spent flying decreased, the slope of metabolic rate on air temperature became more steeply negative, and was significant for bees flying less than 80% of the time. In a separate experiment, there was a significant inverse relationship of metabolic rate and air temperature for bees requiring frequent or constant agitation to remain airborne, but no dependence for bees that flew with little or no agitation; bees were less likely to require agitation during outdoor than indoor measurements. A recent hypothesis explaining differences between studies in the slope of flight metabolic rate on air temperature in terms of differences in metabolic capacity and thorax temperature is supported for honeybees in voluntary flight, but not under agitation.