Reproductive Investment In Response Research Articles

Do non-direct heterospecific cues of avian predator activity alter reproductive modes of a passerine bird?

Predators affect prey by killing them or inducing changes in their physiology and behaviour through a fear effect associated with predation risk. In birds, perceived predation risk influences reproductive decisions, such as the reduction of parental investment in offspring during both egg production and nestling rearing. Visual and vocal cues of predator presence have been widely used to test the direct effects of predation risk. However, few studies have examined the indirect cues of predator activity such as dead avian prey or their remains. In this study, for the first time, we experimentally studied whether piles of feathers, simulating the remains of avian prey, induce changes in the reproductive decisions of adult birds. Before and during egg laying, great tit, Parus major, pairs were exposed to piles of bright down and cover feathers from domestic goose (treatment), woodchips (procedural control), or were not exposed (control). Our experiment affected maternal investment in individual eggs, but did not influence other reproductive parameters. Females from the treatment group laid larger and more asymmetrical (pointed) eggs than control females. Moreover, females from the procedural control group laid larger eggs than those from the control group, but without differences in egg shape. However, the eggs from the treatment and procedural control groups did not differ. This indicates that great tit females can perceive feathers and woodchips as informative cues, such as potential predation risk or habitat suitability, or as novel items in the environment. Importantly, females respond to such cues by changing their maternal investment in eggs, which may result from an adaptive mechanism aimed at increasing offspring fitness in the face of specific environmental conditions experienced by a female. Our study contributes to the understanding of how female songbirds adjust their maternal reproductive investment in response to publicly available social and environmental cues.

Differential allocation in a gift-giving spider: males adjust their reproductive investment in response to female condition

BackgroundWhen males are selective, they can either reject low-quality females or adjust their reproductive investment in response to traits that indicate female quality (e.g., body size or condition). According to the differential allocation hypothesis, males increase their reproductive investment when paired with high-quality females (positive differential allocation) or increase their reproductive investment when paired with low-quality females (negative differential allocation). This hypothesis has been proposed for monogamous species with biparental care, and most empirical studies focus on birds. Here we used the polygamous spider Paratrechalea ornata, in which males offer prey wrapped in silk as nuptial gifts, to test whether males adjust their reproductive investment in gift size, pre-copulatory and copulatory courtship, and sperm transfer in response to female body condition.ResultsMales exposed to females in good body condition added more flies to the gift, stimulated these females longer with abdominal touches during pre-copulatory courtship, and had longer pedipalp insertions than males exposed to females in poor body condition. Female condition affected neither silk investment in nuptial gift wrapping nor the quantity of sperm transferred by males. Finally, females in good body condition oviposited faster after copulation and laid more eggs than females in poor body condition.ConclusionsWe provide experimental evidence that males of a gift-giving spider exhibit positive differential allocation in three key aspects of their reproductive investment: the size of the nutritious gift, duration of pre-copulatory courtship, and duration of pedipalp insertions, which is regarded as a form of copulatory courtship in spiders. This positive differential allocation is likely associated with the benefits of copulating with females in good body condition. These females are more fecund and oviposit faster after copulation than females in poor body condition, which under natural field conditions probably reduces the risk of multiple matings and thus the level of sperm competition faced by the males. As a final remark, our findings indicate that the hypothesis of differential allocation also applies to species with a scramble competition mating system, in which males heavily invest in nuptial gift construction, but not in parental care.

Shifts in Reproductive Investment in Response to Competitors Lower Male Reproductive Success.

AbstractIn many species, males exhibit phenotypic plasticity in sexually selected traits when exposed to social cues about the intensity of sexual competition. To date, however, few studies have tested how this plasticity affects male reproductive success. We initially tested whether male mosquitofish, Gambusia holbrooki (Poeciliidae), change their investment in traits under pre- and postcopulatory sexual selection depending on the social environment. For a full spermatogenesis cycle, focal males were exposed to visual and chemical cues of rivals that were either present (competitive treatment) or absent (control). Males from the competitive treatment had significantly slower-swimming sperm but did not differ in sperm count from control males. When two males competed for a female, competitive treatment males also made significantly fewer copulation attempts and courtship displays than control males. Further, paternity analysis of 708 offspring from 148 potential sires, testing whether these changes in reproductive traits affected male reproductive success, showed that males previously exposed to cues about the presence of rivals sired significantly fewer offspring when competing with a control male. We discuss several possible explanations for these unusual findings.

Limited impacts of chronic eye fluke infection on the reproductive success of a fish host

AbstractParasitic infections may affect the reproductive success of the host either directly, through behavioural modification, or indirectly, by altering their reproductive investment in response to infection. We determined the effects of infection with the eye fluke Diplostomum pseudospathaceum (Trematoda) on the reproductive traits of European bitterling (Rhodeus amarus, Cyprinidae), an intermediate fish host with a resource-based mating system. Male bitterling infected by Diplostomum exhibited a larger but less pronounced red eye spot (sexually selected signal) than control males, suggesting that infected males were less preferred by females. The frequency of female ovulation and number of offspring were comparable between the infected and the control group, although there was a 1–2 week delay in the peak of ovulation and offspring production in infected fish, which is known to coincide with higher juvenile mortality. Chronic eye fluke infection had minimal metabolic costs (measured as oxygen consumption) and, consistent with these results, reproductive activity did not differ between infected and control fish in an experimental test of intersexual selection. Overall, the impact of eye fluke infection on the reproduction of European bitterling was limited. We consider the potential effect of favourable conditions during experiments (abundant food, access to spawning substrate and lack of predators and co-infections) on experimental outcomes and recognize that the effects of chronic eye fluke infection in natural conditions might be more pronounced.

Egg investment in response to helper presence in cooperatively breeding Tibetan ground tits

Life‐history theory predicts a trade‐off between current and future reproduction to maximize lifetime fitness. In cooperatively breeding species, where offspring care is shared between breeders and helpers, helper presence may influence the female breeders’ egg investment, and consequently, survival and future reproductive success. For example, female breeders may reduce egg investment in response to helper presence if this reduction is compensated by helpers during provisioning. Alternatively, female breeders may increase egg investment in response to helper presence if helpers allow the breeders to raise more or higher quality offspring successfully. In the facultatively cooperative‐breeding Tibetan ground tit Pseudopodoces humilis, previous studies found that helpers improve total nestling provisioning rates and fledgling recruitment, but have no apparent effects on the number and body mass of fledglings produced, while breeders with helpers show reduced provisioning rates and higher survival. Here, we investigated whether some of these effects may be explained by female breeders reducing their investment in eggs in response to helper presence. In addition, we investigated whether egg investment is associated with the female breeder's future fitness. Our results showed that helper presence had no effect on the female breeders’ egg investment, and that egg investment was not associated with breeder survival and reproductive success. Our findings suggest that the responses of breeders to helping should be investigated throughout the breeding cycle, because the conclusions regarding the breeders’ adjustment of reproductive investment in response to being helped may depend on which stage of the breeding cycle is considered.

Reproductive investment in Atlantic cod populations off Newfoundland: Contrasting trends between males and females

Life history theory predicts selection for higher reproductive investment in response to increased mortality among mature individuals. We tested this prediction over the period from 1978 to 2013 for three populations of Atlantic cod ( Gadus morhua) off Newfoundland. These populations were heavily fished for a long period. We considered changes in standardized gonad weight as a proxy for changes in gonadal investment. We accounted for the allometry between gonad and body weight, individual body condition, water temperature, and potential spatial and density-dependent effects. Males display significant temporal trends in gonadal investment in all populations; in agreement with theoretical predictions, these trends show increased gonadal investments during the earlier part of the time series when mortality was high, with the trends leveling off or reversing after the later imposition of fishing moratoria. In contrast, females display patterns that are less consistent and expected; significant trends are detected only when accounting for density-dependent effects, with females in two populations unexpectedly showing a long-term decline in gonadal investment. Our results support the hypothesis that fisheries-induced evolution has occurred in gonadal investment in males, but not in females, and suggest that gonadal investment is more important for male reproductive success than expected in this lekking species.

Female reproductive investment in response to male phenotype in wall lizards and its implications for introgression

The likelihood that females will breed or how much they invest in reproduction can depend on the characters of their male partners. Such differential allocation may enhance or limit gene flow between hybridizing lineages, in particular when the lineages have diverged in sexually selected characters. Populations of the common wall lizard (Podarcis muralis) in some regions of Italy exhibit striking exaggeration of coloration, morphology and behaviour compared to populations in France. The two forms hybridize in north-western Italy. Gene flow is directional and in concordance with the prediction that the exaggerated sexual characters of lizards of Italian origin give them a mating advantage. To evaluate if differential allocation contributes to asymmetric introgression, we tested experimentally if female reproductive investment is affected by male origin and male secondary sexual characters. Despite the large genetic and phenotypic divergence between males of Italian and French origin, females did not invest more when paired with males of the same origin, nor when paired with males with highly expressed sexual characters. Combined, these results suggest that female responses to male phenotypes in wall lizards are unimportant for explaining the directional pattern of gene flow in regions of secondary contact.

Male butterflies use an anti‐aphrodisiac pheromone to tailor ejaculates

Summary When females mate with multiple partners, the risk of sperm competition depends on female mating history. To maximize fitness, males should adjust their mating investment according to this risk. In polyandrous butterflies, males transfer a large, nutritious ejaculate at mating. Larger ejaculates delay female remating and confer an advantage in sperm competition. We test whether male ejaculate size in the butterfly Pieris napi (Lepidoptera) varies with female mating history and thus sperm competition, and whether males assess sperm competition using the male‐transferred anti‐aphrodisiac methyl salicylate (MeS) as a cue. Both sexes responded physiologically to MeS in a dose‐dependent manner. Males, however, were more sensitive to MeS than females. Ejaculates transferred by males mating with previously mated females were on average 26% larger than ejaculates transferred by males mating with virgin females, which conforms to sperm competition theory and indicates that males tailored their reproductive investment in response to sperm competition. Furthermore, ejaculates transferred by males mating with virgin females with artificially added MeS were also 26% larger than ejaculates transferred to control virgin females. Male‐transferred anti‐aphrodisiac pheromone not only functions as a male deterrent, but also carries information on female mating history and thus allows males to assess sperm competition.

Exploring simultaneous allocation to mating effort, sperm production, and body growth in male guppies

In several species, males increase their mating and sperm investment in the presence of unfamiliar females, the so-called Coolidge effect. Such an elevated reproductive effort is expected to be associated with a decreased investment in other costly traits, such as somatic growth and maintenance. How precopulatory, postcopulatory, and somatic investments interact one with each other, however, has been rarely evaluated simultaneously. We used the guppy (Poecilia reticulata), a polyandrous livebearing fish with alternative male mating tactics, to compare these 3 investments between males whose encounter rate with unfamiliar females was experimentally maintained at either high (high mate encounter rate [HER]) or low rate (low mate encounter rate [LER]) for 4 months. At the end of this period, HER males showed an increased sperm production as compared with their LER counterparts. This increment was accompanied with a reduction in the time spent following the female and by a shift from costly courtship displays to gonopodial thrusting, a less expensive coercive mating tactic. This effect may indicate a trade-off between these 2 components of male reproductive investment or may reflect a change in male optimal reproductive strategy associated with different mate encounter rate. In contrast, body growth was positively correlated with the increase in sperm production, and did not differ between experimental groups. Collectively, these results suggest that males did not vary their overall reproductive investment in response to their encounter rate with unfamiliar females, but differed significantly in the amount of resources available individually.

Phenotypic plasticity in hormonal and behavioural responses to changes in resource conditions in a migratory songbird

An emerging question in animal behaviour is whether and how behavioural plasticity will enable organisms to adjust to human-induced, rapid environmental changes that affect breeding conditions. Adaptive behavioural plasticity in response to changing resource conditions will depend on the sensitivity of the neuroendocrine system to food stimuli and on constraints in the mechanisms mediating the expression of reproductive behaviours. We tested the hypotheses that food availability mediates plasticity in androgen and corticosterone (CORT) responses, and that circulating concentrations of these hormones in turn mediate investment into competing mating and parental behaviours in a migratory songbird. We provided supplemental food to black-throated blue warblers, Setophaga caerulescens , breeding in habitats with natural low and high food abundance. The effects of supplemental feeding were most pronounced in food-poor habitat. During their social mates' fertile stage, fed males sang less near their nest sites, had lower plasma androgen and CORT levels, and had higher residual mass than control males. Fed males benefited indirectly from their mates' access to supplemental food because fed females provided more parental effort than control females. Fed males did not increase their own parental effort. Our results suggest that fed males increased mate-guarding effort over extrapair mate attraction while their social mates were fertile and invested additional resources into territorial defence or self-maintenance during the parental stage. These findings reveal plastic patterns of covariation among endocrine parameters and mating behaviours, but not parental behaviours, in response to food supplementation. These responses enabled individuals to adjust their reproductive effort rapidly to changes in resource conditions. Our findings suggest that adaptive reproductive investment in response to environmental change is unlikely to be constrained by the hormonal mechanisms underlying the mediation of competing mating and parental behaviours. • Behavioural plasticity can buffer breeding individuals from changes in food resources. • Behavioural plasticity may be constrained by underlying neuroendocrine mechanisms. • Food supplementation increased male mating effort and female parental effort in black-throated blue warblers. • Male behavioural responses to food supplementation covaried with endocrine responses. • Hormone mediation of reproductive investment did not constrain behavioural plasticity.

Demand‐driven resource investment in annual seed production by a perennial angiosperm precludes resource limitation

The limits on annual seed production have long been characterized as restriction by either pollination success or resource provision to seed development. This expected dichotomy between pollen and resource limitation is based on the assumption that reproductive resources are fixed, which is reasonable for semelparous species. In contrast, iteroparity can ease the constraints on reproductive output per breeding season, if resources can be either mobilized from past storage or borrowed against future performance. For perennial plants, these options allow enhanced reproductive investment in response to unusually good pollination, so that annual seed production may not be pollen or resource limited. We assessed demand-governed reproductive investment by manipulating both resource supply capacity (partial defoliation) and resource demand (pollination quality: fully self-pollination, fully cross-pollination, or combinations of partial self- and cross-pollination within the inflorescence) for a forest herb, Stenanthium occidentale, which is subject to strong pre-dispersal inbreeding depression. Insensitivity to partial defoliation indicated that reproductive output was not source regulated. Instead, demand by developing seeds governs resource distribution, as demonstrated by elevated photosynthate translocation to fruits on fully cross-pollinated plants and the ability of completely defoliated plants to produce seeds. Such contingent resource allocation eliminates a simple dichotomy between pollen receipt and resource availability as limits on annual seed production. Instead, such flexible reproductive investment allows iteroparous perennials to participate maximally in current reproduction (as determined by ovule production) following superior pollination, or to conserve resources for future reproduction following poor pollination.

Paternal social enrichment effects on maternal behavior and offspring growth

Paternal environmental experiences are significant predictors of developmental outcomes in offspring and can occur even in the absence of paternal care. Although there has been a recent focus on the role of environmentally induced changes in the male germline in producing these effects, the potential mediating role of mothers has not been investigated. A role for mothers in the transmission of paternal effects has been well acknowledged in behavioral ecology, which predicts that females will dynamically adjust their reproductive investment in response to the qualities of their mate. In the present study, we show that a lifetime of socially enriched compared with impoverished housing conditions shifts anxiety-like behavior and gene expression of male mice. Females that mate with enriched-reared males exhibit increased levels of pup nursing and licking toward their offspring, which are associated with changes in gene expression within the maternal hypothalamus. Significantly, these changes in maternal behavior are correlated with the general levels of anxiety exhibited by their male mates. Further, we show that paternal environmental enrichment results in increased growth of their offspring. These results suggest that maternal-paternal interactions at mating may guide offspring development, with significant implications for the transgenerational transmission of paternal environmental experiences.

Strategic female reproductive investment in response to male attractiveness in birds

Life-history theory predicts that individuals should adjust their reproductive effort according to the expected fitness returns on investment. Because sexually selected male traits should provide honest information about male genetic or phenotypic quality, females may invest more when paired with attractive males. However, there is substantial disagreement in the literature whether such differential allocation is a general pattern. Using a comparative meta-regression approach, we show that female birds generally invest more into reproduction when paired with attractive males, both in terms of egg size and number as well as food provisioning. However, whereas females of species with bi-parental care tend to primarily increase the number of eggs when paired with attractive males, females of species with female-only care produce larger, but not more, eggs. These patterns may reflect adaptive differences in female allocation strategies arising from variation in the signal content of sexually selected male traits between systems of parental care. In contrast to reproductive effort, female allocation of immune-stimulants, anti-oxidants and androgens to the egg yolk was not consistently increased when mated to attractive males, which probably reflects the context-dependent costs and benefits of those yolk compounds to females and offspring.

Risk of Nest Predation Influences Reproductive Investment in American Kestrels (Falco sparverius): an Experimental Test

Nest predation is the primary cause of nest failure in birds. Individuals should therefore adjust parental investment to minimize the costs associated with this constraint; evidence suggests that nest predation influences nest-site selection, and drives variation in both clutch size and parental behavior. Here, we test how the perception of the risk of nest predation from red squirrels (Tamiasciurus hudsonicus) influenced nest-site selection and reproductive investment of American Kestrels (Falco sparverius) breeding in the boreal forest. For this purpose, we conducted audio playbacks of squirrel vocalizations and altered nest boxes to experimentally increase cues of the presence of Red Squirrels in the vicinity of potential nests. Experimental manipulations of the risk of nest predation did not influence nest-site selection; however, experimentally increasing the perceived risk of nest predation induced kestrels to initiate breeding later, and to lay larger clutches. Parents did not appreciably alter incubation behavior in response to our manipulation, although the duration of incubation was longer where natural squirrel threat was higher. Our results showed that kestrels are capable of making facultative adjustments to current reproductive investment in response to their perception of the risk of nest predation.

Sexual Dimorphism in AndrodioeciousMercurialis annua, a Wind-Pollinated Herb

Gender-dimorphic species often display a degree of sexual dimorphism in terms of life-history traits, yet little is known about dimorphism in androdioecious plants. Here we investigate sexual dimorphism in an androdioecious population of the wind-pollinated herb Mercurialis annua by comparing the resource allocation strategies of males and hermaphrodites grown under different nutrient-availability and competitive regimes. We found that males displayed smaller aboveground vegetative sizes than did hermaphrodites, but neither soil nutrient availability nor competition had a strong independent effect on their relative sizes. Plants adjusted their relative reproductive investment in response to nutrient availability. Specifically, hermaphrodites increased their reproductive allocation when growing in poor soils, whereas males displayed the opposite response. Finally, hermaphrodites were strongly female biased in their sex allocation, and this was more pronounced in nutrient-poor soils. To conclude, sexual dimorphism in androdioecious M. annua shares many features with dioecious and gynodioecious species, particularly wind-pollinated herbs. However, the direction of sex-allocation reaction norms displayed by hermaphrodites of M. annua differs from that documented for several insect-pollinated gynodioecious species, hinting at the importance of either the pollination mode or the sexual system as a context of selection shaping the reproductive strategy of plants with both male and female functions.

Temporary loss of a sexual signal (claw loss) affects the frequency of sand structure construction in the fiddler crab, Uca lactea

In iteroparous animals, individual decisions about when and how much to invest in reproduction are related to lifetime reproductive success, and the ability to assess one's own competitive ability in one's current condition is necessary for optimising the reproductive investment. Male fiddler crabs have an enlarged claw which is used for courtship and combat, but they sometimes lose the claw. The clawless males would have disadvantages in courtship and combat until they regenerated the claw. Here it is examined whether males modify their reproductive investment in response to their current condition. Most clawless males did not construct sand structures to attract females; thus, they invested little in current reproduction. Small males were also less likely to construct structures. Clawless and small males may invest little in current reproduction and instead allocate energy to claw regeneration or body growth for future reproduction, or invest in an alternative mating tactic, such as surface mating.

Aphid reproductive investment in response to mortality risks

BackgroundAphids are striking in their prodigious reproductive capacity and reliance on microbial endosymbionts, which provision their hosts with necessary amino acids and provide protection against parasites and heat stress. Perhaps as a result of this bacterial dependence, aphids have limited immune function that may leave them vulnerable to bacterial pathogens. An alternative, non-immunological response that may be available to infected aphids is to increase reproduction, thereby ameliorating fitness loss from infection. Such a response would reduce the need to mount a potentially energetically costly immune response, and would parallel that of other hosts that alter life-history traits when there is a risk of infection. Here we examined whether pea aphids (Acyrthosiphon pisum) respond to immunological challenges by increasing reproduction. As a comparison to the response to the internal cue of risk elicited by immunological challenge, we also exposed pea aphids to an external cue of risk - the aphid alarm pheromone (E)-β-farnesene (EBF), which is released in the presence of predators. For each challenge, we also examined whether the presence of symbionts modified the host response, as maintaining host fitness in the face of challenge would benefit both the host and its dependent bacteria.ResultsWe found that aphids stabbed abdominally with a sterile needle had reduced fecundity relative to control aphids but that aphids stabbed with a needle bearing heat-killed bacteria had reproduction intermediate, and statistically indistinguishable, to the aphids stabbed with a sterile needle and the controls. Aphids with different species of facultative symbiotic bacteria had different reproductive patterns overall, but symbionts in general did not alter aphid reproduction in response to bacterial exposure. However, in response to exposure to alarm pheromone, aphids with Hamiltonella defensa or Serratia symbiotica symbiotic infections increased reproduction but those without a facultative symbiont or with Regiella insecticola did not.ConclusionsOverall, our results suggest that pea aphids are able to increase their reproduction in response to specific cues and that symbiont presence sometimes moderates this response. Such increased reproduction in response to risk of death increases the fitness of both aphids and their vertically transmitted symbionts, and since these organisms have high reproductive capacity, slight increases in reproduction could lead to a very large numerical advantage later in the season. Thus both symbiotic partners can benefit by increasing host fecundity under dangerous conditions.

Females increase reproductive investment in response to helper‐mediated improvements in allo‐feeding, nest survival, nestling provisioning and post‐fledging survival in the Karoo scrub‐robinCercotrichas coryphaeus

In many cooperatively‐breeding species, the presence of one or more helpers improves the reproductive performance of the breeding pair receiving help. Helper contributions can take many different forms, including allo‐feeding, offspring provisioning, and offspring guarding or defence. Yet, most studies have focussed on single forms of helper contribution, particularly offspring provisioning, and few have evaluated the relative importance of a broader range of helper contributions to group reproductive performance. We examined helper contributions to multiple components of breeding performance in the Karoo scrub‐robinCercotrichas coryphaeus, a facultative cooperative breeder. We also tested a prediction of increased female investment in reproduction when helpers improve conditions for rearing young. Helpers assisted the breeding male in allo‐feeding the incubating female, increasing allo‐feeding rates. Greater allo‐feeding correlated with greater female nest attentiveness during incubation. Nest predation was substantially lower among pairs breeding with a helper, resulting in a 74% increase in the probability of nest survival. Helper contributions to offspring provisioning increased nestling feeding rates, resulting in a reduced incidence of nestling starvation and increased nestling mass. Nestling mass had a strong, positive effect on post‐fledging survival. Controlling for female age and habitat effects, annual production of fledged young was 130% greater among pairs breeding with a helper, and was influenced most strongly by helper correlates with nest survival, despite important helper effects on offspring provisioning. Females breeding with a helper increased clutch size, supporting the prediction of increased female investment in reproduction in response to helper benefits.

Do female spotless starlings Sturnus unicolor adjust maternal investment according to male attractiveness?

In birds, female egg allocation patterns have a strong influence in offspring development and differential investment in egg size and composition has been shown to respond to male attractiveness. In this study we experimentally manipulated the perceived attractiveness of male starlings Sturnus unicolor by increasing the amount of green material in some nests (a male courtship display in this species). We predicted that, if female investment before laying is related to male attractiveness, experimental females would increase their reproductive investment in response to the addition of plants in their nests when compared to control females. We found that our manipulation caused variations in female reproductive investment in a way that seems to influence offspring quantity but not offspring quality: Females laid larger clutch sizes but not larger eggs when green plant material was added. However, yolk androgens contents were not related to the experimental manipulation. Contrary to expectations, females breeding in experimental nests laid eggs with smaller amounts of eggshell pigments. Interestingly, we found that eggs laid later in the sequence had higher testosterone levels and showed more intense egg colouration than eggs laid earlier in the sequence. These differences at the within‐clutch level suggest that selection has favoured compensatory strategies for hatching asynchrony. Alternatively, since nest sabotages by other females are most common at the beginning of laying, this could be seen as female strategy to minimise losses due to nest sabotages. As far as we know, this is the first study to show that an external egg characteristic such as blue‐green colouration reflects yolk androgen concentration.

Males use sex pheromone assessment to tailor ejaculates to risk of sperm competition in a butterfly

In polyandrous butterflies males transfer a large, nutritious ejaculate at mating. Larger ejaculates delay female remating and confer an advantage in sperm competition. However, large ejaculates are costly, potentially selecting for male adjustment of ejaculate size to the risk of sperm competition. Here, we test if male ejaculate size in the butterfly Pieris napi varies with male density, and whether males assess sperm competition risk using the male sex pheromone citral as a cue. The results conformed to sperm competition theory and showed that male P. napi tailored their reproductive investment in response to the risk of sperm competition; ejaculates transferred by males in the high male density treatments were on average 23% larger than ejaculates transferred at low male densities. The results also show for the first time that the sex pheromone citral was used by males to assess male density; ejaculates transferred by males in presence of added male sex pheromone were 19% larger than ejaculates transferred in the control. In conclusion, the study shows how the sex pheromone not only facilitates female acceptance when dispensed by courting males but also allows males to assess the degree of male competition for matings.