Female Remating Behavior in a Lekking Moth

In polyandrous species, male reproductive success will at least partly be determined by males' success in sperm competition. To understand the potential for post‐mating sexual selection, it is therefore important to assess the extent of female remating. In the lekking moth Achroia grisella, male mating success is strongly determined by female choice based on the attractiveness of male ultrasonic songs. Although observations have indicated that some females will remate, only little is known about the level of sperm competition. In many species, females are more likely to remate if their first mating involved an already mated male than if the first male was virgin. Potentially, this is because mated males are less well able to provide an adequate sperm supply, nutrients, or substances inhibiting female remating. This phenomenon will effectively reduce the strength of pre‐copulatory sexual selection because attractive males with high mating success will be more susceptible to sperm competition. We therefore performed an experiment designed both to provide a more precise estimate of female remating probability and simultaneously to test the hypothesis that female remating is influenced by male mating history. Overall, approximately one of five females remated with a second male. Yet, although females mated to non‐virgin males were somewhat more prone to remate, the effect of male mating history was not significant. The results revealed, however, that heavier females were more likely to remate. Furthermore, we found that females' second copulations were longer, suggesting that, in accordance with theory, males may invest more sperm in situations with an elevated risk of sperm competition.     

Female Mating History Influences Copulation Behavior but Not Sperm Release in the Orb-Weaving Spider <Emphasis Type="Italic">Tetragnatha versicolor</Emphasis> (Araneae,Tetragnathidae)

We examined the influence of female mating history on copulation behavior and sperm release in the haplogyne spider Tetragnatha versicolor. Despite significant behavioral differences during mating, males released equivalent amounts of sperm to virgin and non-virgin females. When mating with non-virgin females, males showed twice as many pedipalp insertions and half the copulation duration as compared to virgin females; however, males were as likely to mate with non-virgin as virgin females. Even with these overt behavioral differences, males released half of the sperm contained within their pedipalps during mating, regardless of female mating history. With respect to male mating order, first or second, we suggest the numbers of sperm released would lead to an expectation of unbiased paternity. In this species, sperm release is not directly proportional to total copulationduration.     

Sperm competitive ability and genetic relatedness in Drosophila melanogaster: similarity breeds contempt

Abstract.— Offspring of close relatives often suffer severe fitness consequences. Previous studies have demonstrated that females, when given a choice, will choose to avoid mating with closely related males. But where opportunities for mate choice are limited or kin recognition is absent, precopulatory mechanisms may not work. In this case, either sex could reduce the risks of inbreeding through mechanisms that occur during or after copulation. During mating, males or females could commit fewer gametes when mating with a close relative. After mating, females could offset the effects of mating with a closely related male through cryptic choice. Few prior studies of sperm competition have examined the effect of genetic similarity, however, and what studies do exist have yielded equivocal results. In an effort to resolve this issue, we measured the outcome of sperm competition when female Drosophila melanogaster were mated to males of four different degrees of genetic relatedness and then to a standardized competitor. We provide the strongest evidence to date that sperm competitive ability is negatively correlated with relatedness, even after controlling for inbreeding depression.     

Spermatophore displacement and male fertilization success in the ground beetle Carabus insulicola

Sperm displacement is an effective male mating tactic of spermcompetition to ensure paternity. However, the effectivenessof sperm displacement may depend on defensive mating tacticsemployed by rival males and also may be affected by females.To test this hypothesis, I conducted double-mating experimentswith microsatellite paternity analyses using the ground beetleCarabus insulicola. First, I examined how often spermatophoredisplacement by the second male occurs. Spermatophore depositionsuccess of the second male was high (85%) as was that of thefirst male (100%), in which 80% of the second males displacedthe first male's spermatophore, suggesting that spermatophoredisplacement is a highly effective offensive tactic of spermcompetition in C. insulicola. Second, I examined the effectsof postcopulatory guarding by the first male and remating intervaland sperm utilization pattern of the female on fertilizationsuccess of the second male. P₂ values were moderate on average(0.30–0.57) and negatively correlated with postcopulatoryguarding duration of the first male. Sperm use by the femalewas temporally nonrandom, such that sperm of the second maletended to be used first, although there was no significant effectof female-remating interval. These results suggest that theeffectiveness of spermatophore displacement depends on postcopulatoryguarding by the first male and the sperm utilization patternof the female. These findings provide empirical evidence forthe conflicting nature of selection for male offensive and defensivetactics in sperm competition and significant contribution offemales therein.     

Differential sperm expenditure reveals a possible role for post‐copulatory sexual selection in a lekking moth

Male reproductive success in the lesser wax moth Achroia grisella is strongly determined by pre‐copulatory mate choice, during which females choose among males aggregated in small leks based on the attractiveness of ultrasonic songs. Nothing is known about the potential of post‐copulatory mechanisms to affect male reproductive success. However, there is evidence that females at least occasionally remate with a second male and that males are unable to produce ejaculates quickly after a previous copulation. Here we investigated the effects of mating history on ejaculate size and demonstrate that the number of transferred sperm significantly decreased from first (i.e., virgin) to second (i.e., nonvirgin) copulation within individual males. For males of identical age, the number of sperm transferred was higher in virgin than in nonvirgin copulations, too, demonstrating that mating history, is responsible for the decrease in sperm numbers transferred and not the concomitant age difference. Furthermore, the number of transferred sperm was significantly repeatable within males. The demonstrated variation in ejaculate size both between subsequent copulations as well as among individuals suggests that there is allocation of a possibly limited amount of sperm. Because female fecundity is not limited by sperm availability in this system, post‐copulatory mechanisms, in particular sperm competition, may play a previously underappreciated role in the lesser wax moth mating system.     

Complex interactions with females and rival males limit the evolution of sperm offence and defence

Postcopulatory sexual selection favours males which are strong offensive and defensive sperm competitors. As a means of identifying component traits comprising each strategy, we used an experimental evolution approach. Separate populations of Drosophila melanogaster were selected for enhanced sperm offence and defence. Despite using a large outbred population and evidence of substantive genetic variation for each strategy, neither trait responded to selection in the two replicates of this experiment. Recent work with fixed chromosome lines of D. melanogaster suggests that complex genotypic interactions between females and competing males contribute to the maintenance of this variation. To determine whether such interactions could explain our lack of response to selection on sperm offence and defence, we quantified sperm precedence across multiple sperm competition bouts using an outbred D. melanogaster population exhibiting continuous genetic variation. Both offensive and defensive sperm competitive abilities were found to be significantly repeatable only across matings involving ejaculates of the same pair of males competing within the same female. These repeatabilities decreased when the rival male stayed the same but the female changed, and they disappeared when both the rival male and the female changed. Our results are discussed with a focus on the complex nature of sperm precedence and the maintenance of genetic variation in ejaculate characteristics.     

Determinants of male ejaculate investment in the cotton bollworm Helicoverpa armigera: mating history,female body size and male age

Many studies demonstrate that ejaculate size may be influenced by male condition, female quality and the risk or intensity of sperm competition. In the present study, the effect of male and female conditions, male mating history and female mating status on ejaculate sperm numbers in the polyandrous moth Helicoverpa armigera is examined. A large variation in ejaculate size is found and, although female body size and male age influence ejaculate size, female age and copula duration do not. Both male and female mating histories have significant effects on ejaculate sperm numbers. Males reduce ejaculate expenditure in successive matings but deliver significantly more apyrene and eupyrene sperm to nonvirgin than to virgin females.     

Male scorpionflies assess the amount of rival sperm transferred by females' previous mates

Theory predicts that when sperm compete numerically, selection will favor males who vary the number of sperm they transfer with the immediate level of sperm competition. In this study, I measured male mating investment in response to both female mating status (virgin vs. mated) and the number of foreign sperm stored by females in a previous mating in the scorpionfly Panorpa cognata. Female sperm storage was manipulated by interrupting copulations at different time points. Female mating status did not significantly influence male mating investment, but resource-limited males invested strategically in relation to the amount of sperm stored by females in a previous mating. I found continuously decreasing male investment in response to increasing amounts of competing sperm. These results demonstrate an unprecedented male ability to assess the number of sperm stored by females. As a result, males are capable of an extraordinarily fine-tuned reaction to the intensity of sperm competition.     

Paternity costs from polyandry compensated by increased fecundity in the hide beetle

Polyandry-induced sperm competition is assumed to impose costson males through reduced per capita paternity success. In contrast,studies focusing on the consequences of polyandry for femalesreport increased oviposition rates and fertility. For thesespecies, there is potential for the increased female fecundityassociated with polyandry to offset the costs to males of sharedpaternity. We tested this hypothesis by comparing the proportionand number of offspring sired by males mated with monandrousand polyandrous females in the hide beetle, Dermestes maculates,both for males mating with different females and for males rematingwith the same female. In 4 mating treatments, monandrous femalesmated either once or twice with the same male and polyandrousfemales mated either twice with 2 different males or thricewith 2 males (where 1 male mated twice). Polyandrous and twice-matingmonandrous females displayed greater fecundity and fertilitythan singly mating monandrous females. Moreover, males rematedto the same female had greater paternity regardless of whetherthat female mated with another male. In both polyandrous treatments,male mating order did not affect paternity success. Finally,although the proportion of eggs sired decreased if a male matedwith a polyandrous female, multiply mating females or femalesthat remated with a previous mate laid significantly more eggsand thus the actual number of eggs sired was comparable. Thus,males do not necessarily accrue a net fitness loss when matingwith polyandrous females. This may explain the absence of anyobvious defensive paternity-protection traits in hide beetlesand other species.     

Theoretical influence of female mating status and remating propensity on male sperm allocation patterns

Theoretical models predict that males should allocate more sperm in matings where the immediate risk of sperm competition is high. It has therefore often been argued that males should invest less sperm in matings with virgin females compared with matings with already mated females. However, with relatively polyandrous females, high sperm competition risk will covary with high sperm competition intensity leading to more unpredictable conditions, as high competition intensity should favour smaller ejaculates. With the use of a genetic algorithm, we found that males should allocate more sperm in matings with virgin females when female mating frequency is relatively high, whereas low remating rates will select for higher effort in matings with nonvirgin females. At higher remating rates, first male sperm precedence favours larger ejaculates in matings with virgin females and second male precedence favours the reverse. These results shed some light on several findings that have been difficult to explain adaptively by the hitherto developed theory on sperm allocation.     

Female mating status does not affect male mating behavior in the West Indian sweetpotato weevil, Euscepes postfasciatus

Because multiple mating by females encourages sperm competition, the assessment of female mating status before insemination is important for males in order to avoid the risk of sperm competition or to intensify sperm competition. When interacting with females before sperm transfer, males can alter their mating tactics according to the risk or intensity of sperm competition. Information on how mating systems are associated with sperm competition is essential for sterile insect technique eradication programs, which depend on successful mating of released sterile males with wild females. We tested whether males of the West Indian sweetpotato weevil, Euscepes postfasciatus (Fairmaire) (Coleoptera: Curculionidae), adjusted their mating behavior in response to female mating experience and/or age. As virgin/young female weevils accepted males more easily than non-virgin/old females, assessing females before insemination can be adaptive for male weevils. We found that E. postfasciatus males were unable to adjust their mating tactics in response to female mating status. Although this mating strategy would be costly for individual male weevils, the ability in males to discriminate female mating status can prevent the chance of mating with already-mated females containing the sperm of wild male(s). Therefore, the mating tactics of male E. postfasciatus are advantageous for sterile insect technique eradication programs.     

三突伊氏蛛雌蛛不同交配史对雄蛛交配行为的影响

近年来越来越多研究表明,雄性产生精子(精液)也需付出代价。在多次交配的动物中,雄性为获得最大生殖潜力,必须依据配偶的质量策略性地调整每次交配的生殖投入。雄性策略性的生殖投入主要体现在两个方面,一是精子竞争(sperm competition),二是柯立芝效应(Coolidge effect)。目前精子竞争研究主要集中于昆虫类群,而柯立芝效应研究集中于高等脊椎动物,同时验证结果也时常与假说不一致。以多次交配的三突伊氏蛛为材料,以雄蛛交配行为为指标,在蜘蛛类群中探讨和验证雄性精子竞争强度假说和柯立芝效应。设定3个交配组合:2只雄蛛依次与1只雌蛛各交配1次(A组)、2只雄蛛依次与2只雌蛛各交配1次(B组)和1只雄蛛与1只雌蛛交配2次(C组),分析比较3个交配组合的三突伊氏蛛第1次交配和第2次交配在交配潜伏期、交配持续时间和交配回合数方面的差异,比较三突伊氏蛛雌蛛不同交配史对雄蛛交配行为的影响,以此验证雄性精子竞争强度假说和柯立芝效应。研究结果表明A和B组的三突伊氏蛛第2次交配的交配潜伏期和交配持续时间显著长于第1次交配。同时,C组的三突伊氏蛛第1次交配的交配潜伏期和交配持续时间与第2次交配都没有显著差异。同时,A、B和C组的三突伊氏蛛第1次交配的交配回合数与第2次交配都没有显著差异。研究结果支持精子竞争强度假说,而不支持柯立芝效应。     

Male Mating History Influences Female Mate Choice in the Trinidadian Guppy (Poecilia reticulata)

Based on the phenotype‐linked fertility hypothesis, sexual selection should favour females that can accurately assess the recent mating history of available sexual partners and preferentially avoid mating with recently‐mated males [who may be sperm depleted (SD)] so as to minimize the risk of their eggs not being fertilized. This hypothesis has received to date only limited attention and empirical support. Therefore, in the current study, we investigated experimentally whether females of a vertebrate species, the Trinidadian guppy (Poecilia reticulata), are able to assess the recent mating history of males, and thus potentially their functional fertility, and choose to avoid mating with males that appear to have recently mated and who may be sperm limited. Individual virgin females were first given a dichotomous choice between a male that had not been recently observed to interact sexually with another female (i.e. not sperm‐depleted) and another male that had been observed to interact sexually with a female (i.e. potentially sperm‐depleted) as sexual partners. Paired males were matched for body length and coloration. Immediately following this test, the focal females were subjected to a free‐swimming mate‐choice test using the same paired stimulus males. As predicted, on average, female guppies avoided the apparently recently‐mated (and potentially sperm‐depleted) male and exhibited a significant preference for the other male not recently observed mating (and thus not likely sperm limited) during both tests. We do not yet fully understand the underlying mechanisms of this preference. Therefore, further research on the particular cues that females use to assess the recent mating history and fertility status of males is required.     

Old‐male paternity advantage is a function of accumulating sperm and last‐male precedence in a butterfly

Old‐male mating advantage has been convincingly demonstrated in Bicyclus anynana butterflies. This intriguing pattern may be explained by two alternative hypotheses: (i) an increased aggressiveness and persistence of older males during courtship, being caused by the older males' low residual reproductive value; and (ii) an active preference of females towards older males what reflects a good genes hypothesis. Against this background, we here investigate postcopulatory sexual selection by double‐mating Bicyclus anynana females to older and younger males, thus allowing for sperm competition and cryptic mate choice, and by genotyping the resulting offspring. Virgin females were mated with a younger virgin (2–3 days old) and afterwards an older virgin male (12–13 days old) or vice versa. Older males had a higher paternity success than younger ones, but only when being the second (=last) mating partner, while paternity success was equal among older and younger males when older males were the first mating partner. Older males produced larger spermatophores with much higher numbers of fertile sperm than younger males. Thus, we found no evidence for cryptic female mate choice. Rather, the findings reported here seem to result from a combination of last‐male precedence and the number of sperm transferred upon mating, both increasing paternity success.     

Impact of male mating history on the postmating resumption of sexual receptivity and lifetime reproductive success in Choristoneura rosaceana females

Abstract.  The first objective of the present study is to test the hypothesis that the decrease in the number of eupyrene spermatozoa in the spermatheca is directly associated with the resumption of sexual receptivity in female moths, an aspect that has not been examined in previous studies. The obliquebanded leafroller, Choristoneura rosaceana , is used and females mated with previously mated males have a shorter refractory period than those mated with virgins. This difference is associated with a faster rate of movement of sperm from the spermatheca. Overall, the length of the female refractory period coincides with the mean time required for the number of eupyrene sperm in the spermatheca to drop to approximately 3000, regardless of male mating history. Although such a decline in sperm numbers may be a factor responsible for the resumption of sexual receptivity, this is clearly not the only one because more than 40% of females remate even though sperm numbers in the spermatheca are well above this threshold. Virgin males do not vary the mass or the content of their ejaculate as a function of the female's reproductive status and this may increase the risk of sperm competition if the female is previously mated. The second objective of this study is to examine the effect of previous male mating history on female reproductive potential. Females mated with previously mated males have a significantly lower fecundity than those mated with virgin males. However, in all treatments, remating increases both female longevity and lifetime fecundity. There is also a significant effect of female mass on the length of the refractory period and on lifetime fecundity, with large females resuming sexual receptivity sooner and laying more eggs than small ones, regardless of male mating history.     

Determinants of Sperm Transfer by Males of the Noctuid Moth Heliothis virescens

Ejaculate composition can be an important determinant of male reproductive success in the face of sperm competition, which varies with the mating history of the female. Here we examine the effect of various male and female mating histories and morphological traits on ejaculate sperm numbers in the polyandrous moth Heliothis virescens. We show that when mating with nonvirgin females, males passed larger sperm packages (spermatophores) but did not alter either the sperm count or the ratio of nucleated-to-nonnucleated sperm. Males also passed fewer sperm in their second ejaculates. Finally, older males passed more sperm than did younger males. Earlier research found that females store more sperm from older males and that older males are more likely to gain sperm precedence over younger rivals. These earlier results, taken together with the present results, indicate that the advantage enjoyed by older males is due to an increased sperm count.     

Male crickets adjust the viability of their sperm in response to female mating status

Sperm competition theory predicts that males should allocate sperm according to the number of competing ejaculates. Prudent allocation of sperm in response to different levels of sperm competition has been found across a number of taxa; however, some studies suggest that males may not always allocate sperm as expected. Here we examine sperm allocation in the Australian field cricket Teleogryllus oceanicus, using female mating status (virgin, singly mated, or multiply mated) to manipulate male perception of sperm competition risk and intensity. Consistent with theory, we found that male crickets adjust their ejaculates in response to female mating status. However, rather than altering the absolute numbers of sperm transferred to a female, males altered the quality of their sperm. Males ejaculated sperm of low viability (proportion of live vs. dead sperm) when mating with virgins, increased sperm viability when mating with singly mated females, but reduced sperm viability when mating with multiply mated females. Our results show that variation in ejaculate quality can be an important aspect of strategic ejaculation by males and suggest caution in the interpretation of studies in which males do not appear to allocate sperm according to theory.     

Male-derived cuticular hydrocarbons signal sperm competition intensity and affect ejaculate expenditure in crickets

Female sexual promiscuity can have significant effects on male mating decisions because it increases the intensity of competition between ejaculates for fertilization. Because sperm production is costly, males that can detect multiple matings by females and allocate sperm strategically will have an obvious fitness advantage. The presence of rival males is widely recognized as a cue used by males to assess sperm competition. However, for species in which males neither congregate around nor guard females, other more cryptic cues might be involved. Here, we demonstrate unprecedented levels of sperm competition assessment by males, which is mediated via the use of chemical cues. Using the cricket Teleogryllus oceanicus, we manipulated male perception of sperm competition by experimentally coating live unmated females with cuticular compounds extracted from males. We found that males adjusted their ejaculate allocation in response to these compounds: the viability of sperm contained within a male's ejaculate decreased as the number of male extracts applied to his virgin female partner was increased. We further show that males do not respond to the relative concentration of male compounds present on females, but rather to the number of distinct signature odours of individual males. Our results conform to sperm competition theory, and show for the first time, to our knowledge, that males can detect different intensities of sperm competition by using distinct chemical cues of individual males present on females.     

Cryptic female preference for genetically unrelated males is mediated by ovarian fluid in the guppy

As inbreeding is costly, it has been suggested that polyandry may evolve as a means to reduce the negative fitness consequences of mating with genetically related males. While several studies provide support for this hypothesis, evidence of pure post-copulatory mechanisms capable of biasing paternity towards genetically unrelated males is still lacking; yet these are necessary to support inbreeding avoidance models of polyandry evolution. Here we showed, by artificially inseminating a group of female guppies with an equal number of sperm from related (full-sib) and unrelated males, that sperm competition success of the former was 10 per cent lower, on average, than that of the unrelated male. The paternity bias towards unrelated males was not due to differential embryo survival, as the size of the brood produced by control females, which were artificially inseminated with the sperm of a single male, was not influenced by their relatedness with the male. Finally, we collected ovarian fluid (OF) from virgin females. Using computer-assisted sperm analysis, we found that sperm velocity, a predictor of sperm competition success in the guppy, was significantly lower when measured in a solution containing the OF from a sister as compared with that from an unrelated female. Our results suggest that sperm-OF interaction mediates sperm competition bias towards unrelated mates and highlight the role of post-copulatory mechanisms in reducing the cost of mating with relatives in polyandrous females.     

Ejaculate production schedule and the degree of protandry in fishflies (Megaloptera: Corydalidae)

1. To examine the relationship between male–female emergence patterns and ejaculate dynamics, patterns producing sperm and seminal fluids in male internal reproductive organs, the size of a spermatophore transferred at mating and the fate of spermatophore contents moved into female storage organs were compared among the fishflies (Megaloptera: Corydalidae: Chauliodinae), Parachauliodes continentalis, P. japonicus and Neochauliodes sinensis .
2. Spermatophore contents moved into female storage organs decreased rapidly in P. japonicus and N. sinensis , but hardly at all in P. continentalis . This suggests that the females of the former two species may remate sooner than the latter species as it is known in insects that material remaining in the storage organs mechanically inhibits receptivity to mating.
3. Male P. japonicus and N. sinensis increased in internal reproductive organ mass continuously after adult eclosion, and the spermatophore size produced at the first mating increased with male age. In contrast, the internal reproductive organs of P. continentalis were relatively small and did not increase in mass after emergence. P. continentalis transferred a constantly smaller spermatophore at any copulation than the former two species.
4. Males of P. japonicus and N. sinensis emerged earlier than females, while P. continentalis showed a nearly simultaneous emergence pattern between the sexes. It seems that males of P. japonicus and N. sinensis (more polyandrous than P. continentalis ) inhibit female receptivity for a longer time period by emerging earlier and transferring larger ejaculates. In these two species, the number of sperm ejaculated at the first mating also increased with male age. The increase in sperm number by emerging earlier may be adaptive for the males owing to numerical sperm competition when the female remates.