Honeybee defensive behavior is a useful selection criterion, especially in areas with Africanized honeybees (Apis mellifera L). In all genetic improvement programs the selected characters must be measured with precision, and because of this we evaluated a metabolic method for testing honeybee defensive behavior in the laboratory for its usefulness in distinguishing between honeybee ecotypes and selecting honeybees based on their level of defensive responses. Ten honeybee colonies were used, five having been produced by feral queens from a subtropical region supposedly colonized by Africanized honeybees and five by queens from a temperate region apparently colonized by European honeybees. We evaluate honeybee defensive behavior using a metabolic test based on oxygen consumption after stimulation with an alarm pheromone, measuring the time to the first response, time to maximum oxygen consumption, duration of activity, oxygen consumption at first response, maximum oxygen consumption and total oxygen consumption, colonies being ranked according to the values obtained for each variable. Significant (p < 0.05) differences were detected between ecotypes for each variable but for all variables the highest rankings were obtained for colonies of subtropical origin, which had faster and more intense responses. All variables were highly associated (p < 0.05). Total oxygen consumption was the best indicator of metabolic activity for defensive behavior because it combined oxygen consumption and the length of the response. This laboratory method may be useful for evaluating the defensive behavior of honey bees in genetic programs designed to select less defensive bees.

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