Carotenoids in the egg yolks of collared flycatchers (<Emphasis Type="Italic">Ficedula albicollis</Emphasis>) in relation to parental quality,environmental factors and laying order

Birds may influence the fitness of their offspring by transmission of different amounts of carotenoids to their eggs. Carotenoids play crucial roles in antioxidant protection and immune defence mechanisms, but they may be available to females in limiting amounts. Therefore, their allocation to the eggs may be influenced by the female’s condition, age and environmental circumstances. Furthermore, the quality of the male parent, which affects the reproductive value of the offspring, may also influence this investment. In this correlational study, we investigated proximate and ultimate factors that may lead to variation in yolk lutein, zeaxanthin and β-carotene concentrations among and within clutches of a wild passerine, the collared flycatcher (Ficedula albicollis). We found that carotenoid concentration was positively associated with caterpillar supply at the time of egg formation, which suggests a proximate constraint of carotenoid availability on yolk composition. Neither female condition, body size, age, nor male plumage ornamentation, age and body size correlated with carotenoid deposition. Yolk β-carotene concentration was found to be positively linked to yolk testosterone concentration. We suggest that females allocated more β-carotene to their eggs to mitigate the potentially detrimental effects of elevated steroid concentration. We found that concentration of β-carotene increased with laying order. The possible function of this pattern may be to enhance the resistance to oxidative stress and pathogens of the disadvantaged last-hatching nestling, suggesting that collared flycatchers pursue a compensatory, “brood survival” strategy.     

Intra-specific interactions influence egg composition in the lesser black-backed gull (Larus fuscus)

Egg composition, which is under maternal control, can have a profound effect on offspring fitness. The presence of maternal testosterone and carotenoids in avian egg yolk, for example, is thought to enhance the development and competitive ability of the offspring and protect the hatching and growing chick against oxidative stress. Egg quality often differs between females and such variation can be due to differences in maternal social environment, e.g. breeding density. However, this is confounded by the possibility that the quality of individuals breeding in high- or low-density areas may vary. We tested if maternal social environment influences egg composition in a colonial seabird, the lesser black-backed gull (Larus fuscus). To control for confounding effects of female quality, we experimentally manipulated maternal social environment during egg formation. We increased the frequency of intra-specific interactions (i.e. aggressive encounters with conspecifics other than nest mates) in which the females were involved, by placing an elevated platform in their territory. Females that took part in more intra-specific interactions produced a heavier last egg, but the yolk testosterone concentration in eggs laid by control and experimental females did not differ. Differences in yolk testosterone concentration in relation to embryo sex were found neither in the control nor in the experimental group. In contrast, within the control group, eggs with a male embryo contained more carotenoids than eggs with a female embryo. Moreover, experimental females that had been involved in more intra-specific interactions produced female eggs with higher carotenoid levels compared to female eggs of control birds. An experimental increase in carotenoid levels was not observed in eggs containing a male embryo. Our results suggest that intra-specific interactions experienced by female birds during egg formation can influence conditions for embryonic development.Communicated by J. Graves     

Yolk hormones and sexual conflict over parental investment in the pied flycatcher

Female birds might be able to manipulate the parental effort of their male partner through elevated transfer of hormones to the eggs, since these hormones affect many chick traits that males might use as cues for adjusting the level of their investment. We experimentally studied whether female pied flycatchers Ficedula hypoleuca could manipulate male investment via yolk androgens. There is much more variation in yolk androgen levels between females than within clutches, and in order to change the androgen levels of the eggs, we swapped whole clutches between nests. To estimate the androgen levels of the clutch, we measured the androgen content of a single egg per clutch. Females did not succeed in manipulating male effort using yolk androgens, since there was no relationship between the division of parental care within a pair and either original or foster egg androgen levels. One of these relationships should have occurred if females were manipulating males. The proportion of feeding visits by the male was higher when the male was old (55%) than when he was young (45%) and females laid eggs with higher androgen levels when mated with a young male. Young males did not exhibit any responses to yolk androgen levels either, which indicates that females cannot exploit their effort more than that of old males. We suggest that females may allocate yolk androgens to adjust the growth trajectories of the chicks to poor growing conditions when mated with young males that are poor providers or occupying a poor territory.     

Sex-specific effects of yolk-androgens on growth of nestling American kestrels

Maternally derived androgen hormones concentrate in avian egg yolks as the yolks grow on the female’s ovary, possibly forming a basis for important maternal effects in birds. In the American kestrel (Falco sparverius), experimental elevation of yolk androgens in the first-laid egg of a clutch (a-egg) to the concentrations found naturally in a clutch’s later-laid eggs reduces the growth rate of a-egg nestlings compared to controls. These findings, together with discoveries from other species that the effects of yolk androgens on growth of female nestlings may differ from their effects on growth of male nestlings, raise the hypothesis that natural changes in yolk-androgen concentrations with laying order are ultimately due to a difference between the sexes in their yolk-androgen sensitivity and between early- and late-laid eggs in their sex ratio. By re-analyzing previously published data and adding to the analysis data from previously unanalyzed blood samples used for sex determination, we investigated possible sex-specific effects of yolk-androgens in the context of a potential sex-biased laying order in free-living American kestrels. We used a multi-level, mixed model with a Gompertz function to analyze growth of nestlings hatching from a-eggs that were control-treated or in which we experimentally elevated yolk-androgen concentrations shortly after laying to the higher concentrations naturally found in later-laid eggs. We discovered that male nestlings were more susceptible than female nestlings to growth inhibition by yolk-androgen elevation but did not find a bias in sex ratio with respect to laying order. Together, these findings do not support the above hypothesis. However, they are consistent with the hypothesis that sex differences in yolk-androgen sensitivity enable mothers to economically tune reproductive effort to an individual offspring’s reproductive value, which can vary more for one sex than the other.     

Egg pigmentation reflects female and egg quality in the spotless starling <Emphasis Type="Italic">Sturnus unicolor</Emphasis>

It has been proposed that blue colouration in eggs has evolved as a signal of female quality that males can use to modulate their parental investment. This hypothesis is based in the antioxidant properties of biliverdin whose costly deposition in the eggshell is expected to signal female antioxidant capacity and egg quality. Since maternally derived androgens are costly to produce and may adaptively affect offspring phenotype, high-quality females may benefit by signalling their androgen investment through egg colouration. Our aim was to investigate whether egg colour variation in the spotless starling reflected the amount of pigments on the eggshell and whether egg pigmentation was related to female and egg quality. Chromatography analyses revealed that spotless starling eggshells contained two different pigments: biliverdin and protoporphyrin IX with no correlation between them. Biliverdin contents correlated positively with egg colouration indicating that darker eggs with a higher peak in the blue–green segment of the spectrum contained higher amounts of biliverdin. Eggs containing more biliverdin were laid by high-quality females and contained higher yolk testosterone levels. However, despite the strong correlation between biliverdin and colorimetric variables, egg colouration did not reflect accurately female and egg quality. Our results provide evidence that eggshell pigmentation in the spotless starling is related to female and egg quality as shown by the yolk testosterone levels. However, the lack of relation between egg colour and female condition and egg quality do not provide evidence to support the signalling function of egg colouration.     

Are yolk androgens and carotenoids in barn swallow eggs related to parental quality?

Recently, evidence is mounting that females can adaptively engineer the quality of their offspring via the deposition of yolk compounds, including carotenoids and androgens. In this study, we simultaneously consider how both carotenoids and androgens in egg yolk relate to parental quality in barn swallows (Hirundo rustica erythrogaster). First, we found no relationship between concentrations or amounts of yolk androgens and carotenoids. Yolk carotenoids decreased with laying order, whereas we found no relationship between yolk androgens and laying order. Second, we tested the Investment Hypothesis, which predicts that high-quality females or females paired to high quality mates, allocate differentially more of these yolk compounds to their offspring. For carotenoids, we mostly found evidence to counter predictions of the Investment hypothesis: (1) Carotenoid concentrations varied among females, (2) heavier eggs contained lower carotenoid concentrations, although heavier yolks contained greater amounts of carotenoids, (3) eggs of earlier-laying females had lower concentrations in their eggs, and (4) yolk carotenoids were not correlated with clutch size or male plumage ornamentation. For androgens, we found weak support for the Investment Hypothesis: (1) Yolk androgens varied among females, (2) heavier eggs and yolks contained greater amounts, although not concentrations of androgens, (3) females paired to more colorful males laid eggs with greater concentrations of androgens, and (4) no effects of laying date or morphological correlates of female quality on androgen concentrations in egg yolks. Overall, these findings suggest that each yolk compound may have different functions and therefore may be regulated by different mechanisms.     

Androgen-dependent maternal effects on offspring fitness in zebra finches

There is accumulating evidence that maternal hormones may play a role in offspring sex adjustment, but little is known about the costs of such hormone-mediated mechanisms. Recent studies have reported sex-specific effects of hormones on offspring viability. Specifically, we previously found that elevating the plasma androgen level in mothers results in a male-biased offspring primary sex ratio, but it affects the viability of sons negatively and daughters positively in zebra finches (Taeniopygia guttata; Rutkowska and Cichoń, Anim Behav, 71:1283–1288, 2006). In this study, we studied further fitness consequences of exposure to elevated yolk androgen levels in zebra finches. We measured growth rate and cellular immune response of nestlings that hatched from eggs laid by females injected with testosterone during egg laying and nestlings of unaffected control females. We found that sons of testosterone-treated females grew slower in comparison to sons of control females. The significant interaction between experimental group and offspring sex indicates that sons of testosterone-treated mothers suffered impaired immune responsiveness while daughters seemed to benefit from elevated androgen level in terms of enhanced immune responsiveness. We found no effects of androgens on offspring performance at adulthood—neither fecundity of females nor attractiveness of males was affected. We conclude that the benefits of biasing sex ratio towards males by increasing androgen level in the yolk may be limited due to negative effects on male offspring performance early in life.     

Multiple insemination increases reproductive success of female Montandon’s newt (<Emphasis Type="Italic">Triturus montandoni</Emphasis>, Caudata,Salamandridae)

The adaptive significance of multiple matings for females is a matter of much controversy. In insects, supplying the female’s sperm reserves with portions of fresh spermatozoa may be the main function of multiple matings. This simple explanation may also be applied to other animals which produce large numbers of eggs over prolonged periods of time. We tested the fertility insurance hypothesis in Montandon’s newt (Triturus montandoni, Amphibia, Salamandridae). T. montandoni females are inseminated internally by spermatophores they have picked up, and subsequently lay eggs fertilized by spermatozoa released from the spermatheca. We compared the reproductive success of singly and multiply inseminated females of Montandon’s newt in the laboratory. Multiply inseminated females laid more eggs and had a lower percentage of non-developing eggs than females who mated only once. Our data suggest that remating increases the reproductive success of multiply inseminated females by replenishing sperm reserves in the spermatheca or by supplying females with fresh portions of spermatozoa with high fertilizing capacity. Received: 7 January 2000 / Revised: 13 September 2000 / Accepted: 7 October 2000     

Egg coloration in ring-billed gulls (<Emphasis Type="Italic">Larus delawarensis</Emphasis>): a test of the sexual signaling hypothesis

Although many avian eggs appear to be cryptically colored, many species also lay vibrant blue green eggs. This seemingly conspicuous coloration has puzzled biologists since Wallace, as natural selection should favor reduced egg visibility to minimize predation pressure. The sexual signaling hypothesis posits that blue green egg coloration serves as a signal of female quality and that males exert post-mating sexual selection on this trait by investing more in the nests of females laying more intensely blue green eggs. This hypothesis has received mixed support to date, and most previous studies have been conducted in cavity-nesting species where male evaluation of his partner’s egg coloration, relative to that of other females, may be somewhat limited. In this study, we test the sexual signaling hypothesis in colonially nesting ring-billed gulls (Larus delawarensis) where males have ample opportunity to assess their mate’s egg coloration relative to that of other females. We used correlational data and an experimental manipulation to test four assumptions and predictions of the sexual signaling hypothesis: (1) blue green pigmentation should be limiting to females; (2) extent of blue green egg coloration should relate to female quality; (3) extent of blue green egg coloration should relate to offspring quality; and (4) males should provide more care to clutches with higher blue green chroma. Our data provide little support for these predictions of the sexual signaling hypothesis in ring-billed gulls. In light of this and other empirical data, we encourage future studies to consider additional hypotheses for the evolution of blue green egg coloration.     

Female house martins (Delichon urbica) reduce egg androgen deposition in response to a challenge of their immune system

Female birds deposit in the yolks of eggs substantial amounts of androgens, such as testosterone and androstenedione. These androgens have been shown to speed up nestling development, induce a fast development of ornaments and increase dominance in adults. Experiments in several species have reported that females invest greater amounts of androgens in the eggs fathered by attractive males, suggesting that yolk androgen is a costly investment for either the offspring or the mother. There is some evidence that nestling immunocompetence may be partially suppressed by high levels of yolk androgens, but it is not known whether this is also the case for females. We tested this hypothesis in the house martin by inducing an immune challenge through an injection of sheep red blood cells, a standard challenge of the humoral immune system. Experimental birds laid eggs with lower amounts of yolk androstenedione than controls, and there was a similar non-significant trend for testosterone. Furthermore, the probability of laying a replacement clutch was higher for birds that had laid a first clutch with relatively high levels of yolk testosterone. These results suggest that yolk androgen deposition is limited by immune costs in the female, and that only females in good condition may afford to invest high levels of androgen in eggs in this species.     

Maternal nutrition affects offspring performance via maternal care in a subsocial spider

Provisioning of nutrients to the young during parental care is one of the mechanisms by which parents can affect growth and survival of their young and thus their reproductive success. We examined the hypothesis that food quality, i.e., ratio of macronutrients, provided to the young via maternal care affects their performance and the female’s reproductive success. The subsocial spider Stegodyphus lineatus exhibits intensive maternal care behaviors, including feeding the young and matriphagy (consumption of the mother). Our results showed that a protein-enriched diet resulted in larger females at maturation and higher survival of young, relative to intermediate or lipid-enriched diets. However, fecundity was not affected by female diet. We suggest that most of the nutrients are provided to the young during maternal care rather than deposited in the eggs, allowing females to economize on limited nutritional reserves. Females before maternal care showed a low protein and high lipid content relative to females before maturation and oviposition, suggesting a change in the nutritional requirement of females before maternal care. This change in macronutrient composition may be adaptive for the success of the young in the wild and shows a novel approach to animals’ ability to increase their reproductive success. Field-collected females showed a similar macronutrient composition as that of protein-enriched females in the lab, suggesting a high reproductive success of females in the wild. To our knowledge, this is the first time the importance of different macronutrients to females’ reproductive success is examined during reproduction.     

Variability in larval development of the tropical reef fish Pomacentrus amboinensis (Pomacentridae): the parental legacy

The effects of food availability, female size, and social interactions on the quality of Pomacentrus amboinensis larvae at hatching were examined using two field-based experiments. In Experiment 1, food availability and female size significantly influenced size, eye diameter and levels of yolk reserves of larvae at hatching. Small females (47 to 52 mm standard length, SL) whose diets were not supplemented, produced the longest larvae (3.0 ± 0.01 mm total length, TL) with the least yolk reserves (50.1 ± 1.04 μm²). Irrespective of female size, those that received additional food produced larvae with the largest yolk-sacs (large females: 87.60 ± 1.53 μm²; small females: 80.14 ± 1.24 μm²). In Experiment 2, interactions with conspecifics had a greater affect on the somatic development of larvae at hatching than food availability. Increased social interactions resulted in larvae that were ⋍3% longer, with 2% greater head depth, than larvae from females that spawned in isolation on the experimental reefs. Fed females produced larvae with ⋍20% more yolk than larvae from females whose diets were not supplemented. All three factors (food availability, female size, and intensity of social interactions) tested within these experiments vary spatially and temporally among reefs. There is the potential, therefore, for larvae at the onset of the planktonic stage to vary in quality, level of development, and probability of survival. Received: 12 August 1996 / Accepted: 26 August 1996     

Exogenous testosterone increases female aggression in the European starling (<Emphasis Type="Italic">Sturnus vulgaris</Emphasis>)

In the European starling, Sturnus vulgaris, optimal mating systems differ between males and females. Males gain from polygyny, whereas monogamy increases female fitness. The cost of polygyny to females lead to intense female–female competition, and it has previously been shown that the intensity of female aggression during the pre-breeding period can predict the realised mating system. The physiological regulation of such female aggression in starlings is not yet known. This study examines the role of testosterone in mediating aggressive behaviours involved in intra-specific reproductive competition in female starlings. Testosterone levels were experimentally elevated with testosterone implants in females during the pre-laying period. To simulate a situation in which an additional female tried to mate with the focal female’s mate, a caged female was presented close to a nest-site to which the male could attract a secondary female. Testosterone was significantly related to several behaviours involved in female–female interactions. Females with testosterone implants spent significantly more time close to the caged female and produced more song bouts than control females. In contrast, male behaviour was unrelated to the experimental status of the mate. Females mated to males that attracted a secondary female were less aggressive towards the caged female than those that remained monogamously mated. The effect of exogenous testosterone in this study indicates that androgens may mediate social behaviours in female starlings during the breeding season.     

Mothers matter: crowding leads to stressed mothers and smaller offspring in marine fish

Most marine populations are sustained by the entry of juveniles that have survived the larval phase, during which time most die. The number of survivors depends strongly on the quality of the eggs produced by spawning females, but it is not known how the social conditions under which breeding occurs influence the quality of larvae produced. Here I show that the density of females interacting with breeding mothers directly influences the size of larvae produced, through a stress-related mechanism. On the Great Barrier Reef of Australia, breeding pairs of a damselfish, Pomacentrus amboinensis, were isolated on habitat patches, and additional females that could not access the spawning site were added at four densities (0, 1, 3, or 6 females). Additional females increased aggressive interactions by mothers and increased the levels of the stress hormone, cortisol, in their ovaries, leading to reduced larval size. Neither egg output nor yolk size of the larvae was influenced by female density. Pairs breeding in isolation produced the largest larvae; current theory suggests that these larvae should contribute most to subsequent population replenishment events. This social mechanism may influence which females effectively contribute to the next generation and may promote resilience in patchy or isolated populations.     

Production, hatching success and surface ornamentation of eggs of calanoid copepods during a winter at 57°N

Close to 50 species of marine Calanoid copepods have been reported to produce diapause eggs (Engel and Hirche in J Plankton Res 26:1083–1093, 2004); eggs that are viable but require a refractory phase before they hatch, sometimes after months. Diapause eggs are often described as morphologically different with respect to egg membrane ultrastructure and having a thicker egg shell with surface ornamentation as opposed to the smooth shell found in subitaneous eggs that hatch within days (Belmonte in J Mar Syst 15:35–39, 1998; Chen and Marcus in Mar Biol 127:587–597, 1997; Castro-Longoria in Crustaceana 74:225–236, 2001). Egg production rates, egg surface ornamentation, and hatching success were monitored in large aquaculture fish enclosures during winter with close to zero water temperatures (N57°). Surprisingly, all female copepods (Acartia spp.—presumably A. tonsa, and Centropages hamatus) produced eggs all through the winter with no obvious pattern with respect to light, temperature and food availability, and no diapause eggs were observed. However, individual females produced several categories of eggs with or without surface spines even within the same egg batch as evidenced by scanning electron microscopy (SEM). Four egg categories were distinguishable: ‘no spines’, smooth eggs; ‘short spines’, 5–15 μm long; ‘truncated spines’, with the spine tips cut-off <10 μm long; and ‘long spines’, up to 30 μm long. All egg categories remained unchanged with respect to surface structures from when we took them out of the incubation bottles until they hatched. In general, the frequency of ‘no spines’ was 10–40%, and most eggs were ornamented with ‘short-’ or ‘long spines’. Further, a given egg can be ornamented with all types of surface spines simultaneously, which might even be a fifth egg category. The different egg categories were all able to hatch within days when exposed to normoxic conditions suggesting that they were subitaneous.     

Pup production,sex ratios,and survivorship in African wild dogs, <Emphasis Type="Italic">Lycaon pictus</Emphasis>

The local resource enhancement (LRE) model predicts that in cooperatively breeding species, sex ratios will be biased in favor of the more helpful sex. In this study, we assess the assumptions underlying the LRE model in a population of cooperatively breeding wild dogs (Lycaon pictus) in Northern Botswana monitored over a 15-year period. In this population, litter size and pup survival to 1 year are strongly affected by pack size and the breeding female’s age, but adult males have a stronger and more linear effect on females’ reproductive performance than do adult females. This asymmetry in the benefits derived from male and female helpers is reflected in male-biased sex ratios in litters at the time pups emerge from the den. Sex ratio biases are most pronounced in the litters of the youngest mothers who live in significantly smaller packs than older females. The presence of potential rivals for the dominant female’s position depresses pup production at the time of emergence, suggesting that competition among females for breeding positions may also contribute to the selective forces affecting birth sex ratios.     

Tug-of-war over reproduction in a cooperatively breeding cichlid

In group-living animals, dominants may suppress subordinate reproduction directly and indirectly, thereby skewing reproduction in their favour. In this study, we show experimentally that this ability (‘power’) is influenced by resource distribution and the body size difference between unrelated dominants and subordinates in the cichlid Neolamprologus pulcher. Reproduction was strongly skewed towards the dominant female, due to these females producing more and larger clutches and those clutches surviving egg eating better than those of subordinate females, but was not so when subordinates defended a patch. If breeding shelters were provided in two patches, subordinate females were more likely to exclusively defend a patch against the dominant female and breed, compared to when the same breeding resource was provided in one patch. Relatively large subordinate females were more likely to defend a patch and reproduce. Females also directly interfered with each other’s reproduction by eating the competitors’ eggs, at which dominants were more successful. Although dominant females benefited from subordinate females due to alloparental care and an increase in egg mass, they also showed costs due to reduced growth in the presence of subordinates. The results support the view that the dominant’s power to control subordinate reproduction determines reproductive partitioning, in agreement with the predictions from tug-of-war models of reproductive skew.     

Long-lasting effects of yolk androgens on phenotype in the pied flycatcher (Ficedula hypoleuca)

The hormonal environment during early development, such as maternally derived androgens in bird eggs, shapes the development of the offspring in ways that may have important long-term consequences for phenotype and behavior and, ultimately, fitness. We studied the long-term effects of yolk androgens on several phenotypic and physiological traits in male and female pied flycatchers (Ficedula hypoleuca) by experimentally elevating yolk androgen levels and rearing birds in common-garden environment in captivity. We found that high yolk androgen levels increased the basal metabolic rates in both females and males in adulthood. High yolk androgen levels did not affect male melanin coloration or plumage ornaments, or timing or speed of moult in either sex. No effect of androgen treatment on cell-mediated or humoral immune response was found in either sex. Covariation among the measured phenotypic traits was further not altered by androgen treatment. Our results suggest that exposure to high androgen levels can have long-lasting effects on some offspring traits, but do not seem to lead to different phenotypes. Furthermore, the role of yolk androgens affecting sexually selected male traits in our study species seems to be minor. The fitness consequences of yolk androgen-induced higher metabolic rates remain to be studied.     

Sex ratio varies with egg investment in the red-necked phalarope (Phalaropus lobatus)

Fisher’s sex ratio theory predicts that on average parents should allocate resources equally to the production of males and females. However, when the cost/benefit ratio for producing males versus females differs, the theory predicts that parents may bias production, typically through underproduction of the sex with greater variation in fitness. We tested theoretical predictions in the red-necked phalarope, a polyandrous shorebird with sex-role reversal. Since females are larger and therefore potentially more expensive to produce and may have greater variation in reproductive success, we predicted from Fisher’s hypothesis a male bias in population embryonic sex ratio, and from sex allocation theory, female biases in the clutches of females allocating more resources to reproduction. We measured eggs and chicks and sexed 535 offspring from 163 clutches laid over 6 years at two sites in Alaska. The embryonic sex ratio of 51.1 M:48.9 F did not vary from parity. Clutch sex ratio (% male) was positively correlated with clutch mean egg size, opposite to our prediction. Within clutches, however, egg size did not differ by sex. Male phalarope fitness may be more variable than previously thought, and/or differential investment in eggs may affect the within-sex fitness of males more than females. Eggs producing males were less dense than those producing females, possibly indicating they contained more yolk relative to albumen. Albumen contributes to chick structural size, while yolk supports survivorship after hatch. Sex-specific chick growth strategies may affect egg size and allocation patterns by female phalaropes and other birds.     

Polyandry, multiple mating, and female fitness in a water strider Aquarius paludum

Contrary to Bateman’s principle, polyandry appears to be a common female mating strategy. Several hypotheses have been proposed to explain the evolution of polyandry. It is assumed that females gain either material or genetic benefits from multiple matings, or that they are coerced into mating by males. In water striders, mating is generally assumed to be costly to females, and they are thought to mate for reasons of convenience, adjusting their resistance to mating according to male harassment. Here, we tested the effect of number of matings (with the same male) and number of partners on female fitness in a water strider Aquarius paludum. In the first experiment, we regulated the time females spent with a male and found that females’ egg production increased with multiple matings up to a point. The result supports the existence of an optimal female mating frequency. In the second experiment, we tested how polyandry affects the number of eggs laid and egg hatching success. We conducted three different trials: females mated four times with either a single male, two different males, or with four different males. Females that mated with four different males laid the lowest number of eggs and had the lowest egg hatching success, suggesting that polyandry reduces females’ egg production and egg hatching success in A. paludum. We conclude that A. paludum females probably gain material benefits from mating but no genetic benefits were found in this study.