Visual and olfactory associative learning in the malaria vector Anopheles gambiae sensu stricto

Background: Memory and learning are critical aspects of the ecology of insect vectors of human pathogens because of their potential effects on contacts between vectors and their hosts. Despite this epidemiological importance, there have been only a limited number of studies investigating associative learning in insect vector species and none on Anopheline mosquitoes. Methods: A simple behavioural assays was developed to study visual and olfactory associative learning in Anopheles gambiae, the main vector of malaria in Africa. Two contrasted membrane qualities or levels of blood palatability were used as reinforcing stimuli for bi-directional conditioning during blood feeding. Results: Under such experimental conditions An. gambiae females learned very rapidly to associate visual (chequered and white patterns) and olfactory cues (presence and absence of cheese or Citronella smell) with the reinforcing stimuli (bloodmeal quality) and remembered the association for up to three days. Associative learning significantly increased with the strength of the conditioning stimuli used. Importantly, learning sometimes occurred faster when a positive reinforcing stimulus (palatable blood) was associated with an innately preferred cue (such as a darker visual pattern). However, the use of too attractive a cue (e. g. Shropshire cheese smell) was counterproductive and decreased learning success. Conclusions: The results address an important knowledge gap in mosquito ecology and emphasize the role of associative memory for An. gambiae's host finding and blood-feeding behaviour with important potential implications for vector control.