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Background  Sexual selection theory predicts that females, being the limiting sex, invest less in courtship signals than males. However, when chemical signals are involved it is often the female that initiates mating by producing stimuli that inform about sex and/or receptivity. This apparent contradiction has been discussed in the literature as 'the female pheromone fallacy'. Because the release of chemical stimuli may not have evolved to elicit the male's courtship response, whether these female stimuli represent signals remains an open question. Using techniques to visualise and block release of urine, we studied the role of urine signals during fighting and mating interactions of crayfish ( Pacifastacus leniusculus ). Test individuals were blindfolded to exclude visual disturbance from dye release and artificial urine introduction.    Results  Staged female-male pairings during the reproductive season often resulted in male mating attempts. Blocking female urine release in such pairings prevented any male courtship behaviour. Artificial introduction of female urine re-established male mating attempts. Urine visualisation showed that female urine release coincides with aggressive behaviours but not with female submissive behaviour in reproductive interactions as well as in intersexual and intrasexual fights. In reproductive interactions, females predominately released urine during precopulatory aggression; males subsequently released significantly less urine during mating than in fights.    Conclusions  Urine-blocking experiments demonstrate that female urine contains sex-specific components that elicit male mating behaviour. The coincidence of chemical signalling and aggressive behaviour in both females and males suggests that urine release has evolved as an aggressive signal in both sexes of crayfish. By limiting urine release to aggressive behaviours in reproductive interactions females challenge their potential mating partners at the same time as they trigger a sexual response. These double messages should favour stronger males that are able to overcome the resistance of the female. We conclude that the difference between the sexes in disclosing urine-borne information reflects their conflicting interests in reproduction. Males discontinue aggressive urine signalling in order to increase their chances of mating. Females resume urine signalling in connection with aggressive behaviour, potentially repelling low quality or sexually inactive males while favouring reproduction with high quality males.

To signal or not to signal? Chemical communication by urine-borne signals mirrors sexual conflict in crayfish