Sperm competition is expected to favour the evolution of traits that influence the performance of sperm when they compete to fertilize a female's eggs. While there is considerable evidence that selection favours increases in sperm numbers, much less is known about how sperm quality contributes towards competitive fertilization success. Here, we determine whether variation in sperm quality influences competitive fertilization success in the green swordtail Xiphophorus helleri, a highly promiscuous livebearing fish. We use artificial insemination as a method of controlled sperm delivery and show that sperm swimming velocity is the primary determinant of fertilization success when ejaculates from two males compete to fertilize a female's eggs. By contrast, we found no evidence that sperm length had any effect on siring success. We also found no evidence that pre- and postcopulatory sexual traits were phenotypically integrated in this species, suggesting that the previous observation that reproductive skew favours males with high mating rates is unlikely to be due to any direct association between sperm quality and male sexual ornamentation.

Multiple mating is thought to provide an opportunity for females to avoid the costs of genetic incompatibility by postcopulatory selection of compatible sperm haplotypes. Few studies have tested the genetic incompatibility hypothesis directly. Here we experimentally manipulated the compatibility of females with their mates using the gryllid cricket Teleogryllus oceanicus. We recorded the hatching success of eggs laid by females mated with two nonsibling males, two siblings, or one nonsibling male and one sibling. In contrast with two previous studies on crickets that have adopted this approach, the hatching success of eggs did not differ between females mated with two full siblings and females mated with two unrelated males, indicating that embryo viability was not a cost of inbreeding in this species. We assigned paternity to offspring produced by females mated to both a sibling and a nonsibling male using microsatellite markers. As in previous studies of this species, we were unable to detect any difference in the proportion of offspring sired by the 1st and the 2nd male to mate with a female when females were unrelated to their mates. However, in our experimental matings the proportion of offspring sired by the nonsibling male depended on his sequence position. Paternity was biased toward the nonsibling male when he mated first. Our data show that molecular analyses of paternity are essential to detect subtle mechanisms of postcopulatory sexual selection.

Empirical tests of sexual selection theory generally utilize model systems under laboratory settings, and extend conclusions to evolutionary processes occurring in nature. The biological significance of laboratory findings will depend largely on the mating rates of females and patterns of paternity in natural populations, information on which is generally lacking. Here we use microsatellite markers to provide rare estimates of female mating rates and patterns of parentage in a species of tettigoniid, Requena verticalis, which has been used extensively to test theory on the evolution of male parental investment and its influence on the direction of sexual selection. We found that although the number of males having a genetic representation in the female's sperm stores was higher for females collected late in the breeding season than those collected early in the season, overall the female mating rate was lower than that expected from laboratory observations. Analysis of parentage of offspring produced by females at the end of the breeding season revealed that all males represented in the sperm stores fathered offspring, although paternity was biased away from that expected from random sperm utilization. The data show that the complete first male sperm precedence documented in laboratory studies of this species does not persist in natural populations. Our data provide a solid underpinning for conclusions drawn from laboratory studies of this species.

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Recent evidence shows that females exert a post-copulatory fertilization bias in favour of unrelated males to avoid the genetic incompatibilities derived from inbreeding. One of the mechanisms suggested for fertilization biases in insects is female control over transport of sperm to the sperm-storage organs. We investigated post-copulatory inbreeding-avoidance mechanisms in females of the cricket Teleogryllus oceanicus. We assessed the relative contribution of related and unrelated males to the sperm stores of double-mated females. To demonstrate unequivocally that biased sperm storage results from female control rather than cryptic male choice, we manipulated the relatedness of mated males and of males performing post-copulatory mate guarding. Our results show that when guarded by a related male, females store less sperm from their actual mate, irrespective of the relatedness of the mating male. Our data support the notion that inhibition of sperm storage by female crickets can act as a form of cryptic female choice to avoid the severe negative effects of inbreeding.

Recent studies have suggested that males may vary the quality of their ejaculates in response to sperm competition, although the mechanisms by which they do so remain unclear. The viability of sperm is an important aspect of ejaculate quality that determines competitive fertilization success in the field cricket Teleogryllus oceanicus. Using in vitro mixtures of sperm and seminal fluid from pairs of male crickets, we show that seminal fluid can affect the viability of sperm in this species. We found that males who invest greatly in the viability of their own sperm can enhance the viability of rival sperm, providing the opportunity for males to exploit the investments in sperm competition made by their rivals. Transitive effects of seminal fluids across the ejaculates of different males are expected to have important implications for the dynamics of male investments in sperm competition.

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