Can males detect the strength of sperm competition and presence of genital plugs during mate choice?

Female reproductive status can have strong selective effects on male mating strategies due to the threat of sperm competition, which may explain why males prefer virgin over mated females. However, in mating systems with female multiple mating rates and mating plugs, males should not only respond to the risk but also to the level of sperm competition and should be sensitive to the interference from mating plugs. In the orb-web spider Argiope keyserlingi, females possess paired sperm-storage organs facilitating separate sperm storage from different males. Males are limited to 2 copulations due to mutilation of their paired genitals (pedipalps). By conducting binary choice experiments, we tested whether males of different mating status can distinguish between females that mated with 1 or 2 males and whether single-mated males discriminate between single-mated females with matched or unmatched virgin genital openings. Furthermore, we investigated whether males adapt their mating strategies to the intensity of sperm competition by providing males with varying qualities of females in their immediate vicinity. Our results demonstrated that males are sensitive to the level of sperm competition and preferred single-mated females over double-mated females. However, they failed to identify single-mated females whose virgin genital opening matched their unused pedipalp, which is required due to their fixed ipsilateral insemination. Surprisingly, males never mated twice with the same female independently of the quality of surrounding females. This suggests that the benefits from searching and mating with a different female are greater than the benefits of monopolizing the female by mating twice with her.