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Copulation does not occur in the absence of this chemical display.
In further contrast to D. melanogaster, where seminal fluid increases egg laying behaviour but shortens female lifespan [ 22, 22], in S. cynipsea copulation does not appear to enhance egg laying [ 38] or be otherwise toxic for females, although in another study longer copulations did reduce the number of offspring a female produced [ 52].
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The number of sperm present 1 and 3 days after copulation did not differ in the SEV (t16 = 1.28, p = 0.22), but did differ in the SPV (t16 = 2.92, p = 0.010).
The number of sperm present 1 and 3 days after copulation did not differ significantly in the SEV (t14 = 0.88, p = 0.39) or the SPV (t14 = 1.06, p = 0.36).
As a negative control, we attempted to identify unlabeled proteins from a female that had undergone N labeling for three weeks and was paired with a male for approximately 20 hours, but where copulation did not take place, as confirmed by the absence of a copulatory plug.
Genital licking was observed in 14 of 20 copulations, whereas six copulations did not involve licking behavior (Fig. 1).
Interrupted copulations do not result in sperm transfer, although they may lead to the transfer of seminal fluid [40].
The mean durations of copulations did not significantly differ between the four trials (ANOVA on trials 1 to 4, male ID as random effect: F3,59 = 2.47, p = 0.074).
Instead, we suggest that the observed patterns are most consistent with a fertility insurance role for extra-pair copulations, which does not exclude the greater opportunity for sexual selection through differential ability of males to gain paternity.
Many times a successful copulation and ejaculate transfer does not result in a female laying any eggs (i.e., a successful copulation with the female having laid few eggs despite many in her abdomen).
If the male-male competition hypothesis does not explain copulation calling behaviour, then why do females call?
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