a major role in host-seeking behaviour in haematophagous insects (Lazzari, 2009;Guerenstein and Lazzari, 2009 Accepted 28 June 2011 SUMMARY It has been largely assumed that the individual experience of insects that are disease vectors might not only contribute to animal fitness, but also have an important influence on parasite transmission. Nevertheless, despite the invested efforts in testing the capacity to learn and remember information in blood-sucking insects, only little conclusive information has been obtained to date. Adapting a classical conditioning approach to our haematophagous model, we trained larvae of Rhodnius prolixus to associate Llactic-acid, an odour perceived by these bugs but behaviourally neutral when presented alone, with food (i.e. positive reinforcement). Naive bugs -those exposed either to a conditioned stimulus (CS, L-lactic acid), unconditioned stimulus (US, heat) and reward (blood) alone or CS, US and reward in the absence of contingency -remained indifferent to the presence of an air stream loaded with L-lactic acid when tested in an olfactometer (random orientation), whereas the groups previously exposed to the contingency CS-US-reward (blood) were significantly attracted by L-lactic-acid. In a companion paper, the opposite, i.e. repellence, was induced in bugs exposed to the contingency of the same odour with a negative reinforcement. This constitutes the first evidence of olfactory conditioning in triatomine bugs, vectors of Chagas disease, and one of the few substantiations available to date of olfactory conditioning in haematophagous insects.

SUMMARYAfter having demonstrated that blood-sucking bugs are able to associate a behaviourally neutral odour (L-lactic acid) with positive reinforcement (i.e. appetitive conditioning) in the first part of this study, we tested whether these insects were also able to associate the same odour with a negative reinforcement (i.e. aversive conditioning). Learned aversion to host odours has been repeatedly suggested as a determinant for the distribution of disease vectors among host populations. Nevertheless, no experimental evidence has been obtained so far. Adapting a classical conditioning approach to our haematophagous model, we trained larvae of Rhodnius prolixus to associate L-lactic acid, an odour perceived by bugs but behaviourally neutral when presented alone, with a mechanical perturbation (i.e. negative reinforcement). Naive bugs and bugs exposed to CS, punishment, or CS and punishment without contingency remained indifferent to the presence of an air stream loaded with L-lactic acid (random orientation on a locomotion compensator), whereas the groups previously exposed to the contingency CS-punishment were significantly repelled by L-lactic acid. In a companion paper, the opposite, i.e. attraction, was induced in bugs exposed to the contingency of the same odour with a positive reinforcement. These constitute the first pieces of evidence of olfactory conditioning in triatomine bugs and the first demonstration that the same host odour can be used by insects that are disease vectors to learn to recognize either a host to feed on or a potentially defensive one. The orientation mechanism during repulsion is also discussed in light of our results.