In learning and memory studies on honeybees ( Apis mellifera
), cold-induced narcosis has been widely used to temporarily immobilize honeybees. In this study, we investigated the effects of cold narcosis on the associative memories in honeybees by using the proboscis extension response (PER) paradigm. Severe impairments in memory acquisition was found when cold narcosis was performed 30 min, instead of 1 h before training. Locomotor activities were reduced when honeybees were tested 15 min, instead of 30 min after cold narcosis. These results indicate that cold narcosis impairs locomotor activities, as well as memory acquisition in a time-dependent manner, but by comparison no such effects on memory retrieval have yet been observed.