Single Copulation Research Articles

Copulation Duration and Sperm Precedence with Reference to Larval Diapause Induction in Monochamus alternatus Hope (Coleoptera: Cerambycidae).

Adults of the pine sawyer Monochamus alternatus are the primary vector of Bursaphelenchus xylophilus, the causative agent of pine wilt disease. A sawyer subspecies in Taiwan (abbreviated 'T') has two generations a year (bivoltinism) due to facultative diapause, whereas another subspecies in Japan (abbreviated 'J') has a one- or two-year life cycle due to obligate diapause. T, with two infection periods a year, will cause more severe disease epidemics than J if it is introduced into Japan. Inter-subspecies hybridization may inhibit the expression of bivoltinism because many F1 hybrids induce diapause. To predict the effects of introducing T into Japan, the present study investigated copulation duration and late-male sperm precedence to fertilize eggs. The results indicated that a single copulation for more than 65 s supplied sufficient sperm to fertilize a lifetime production of eggs. The incidence of larval diapause was 0.15 for the offspring of T females that mated with a T male and increased to 0.292-0.333 after remating with a J male, while the incidence of larval diapause was 0.900-1.000 for hybrids from T females mated with a J male. Consequently, the estimated proportion of second-male sperm used by T females was 0.185-0.217. The effects of introducing T populations into Japan on the severity of disease epidemics were also discussed.

Mating roles, copulation duration and reproductive output in the hermaphroditic freshwater snail Planorbella trivolvis

ABSTRACT Hermaphrodites exhibit a wide variety of reproductive strategies, yet empirical studies lag behind theories about their evolution. Planorbella trivolvis, a simultaneous hermaphrodite capable of reciprocal copulation and rare self-fertilization, provides a unique model system for studying reproduction. This study assessed sex roles during mating, demonstrating that mating is primarily reciprocal, and investigated the relationship between physical coupling and reproductive output for single copulation events. Snails were isolated before sexual maturity and placed in 10-l aquaria in size-matched groups. Upon copulation, pairs were removed to small cups until they separated; they were then kept isolated and eggs counted until egg production ceased. The frequency of reciprocal copulation (both members of a pair produced egg masses following copulation) was 95% (35/37 productive matings), higher than previously reported. The duration of egg production ranged from 19 to 197 days (mean ± SE = 110.13 ± 6.3 days) with a sharp drop in production of egg masses and eggs per mass occurring between 15 and 20 weeks postmating, consistent with previous estimates of sperm storage. There was no relationship between mating duration and total egg production (a biomarker of sperm availability) or the duration of egg production (a marker of sperm longevity). Further investigation is needed to directly estimate sperm transfer and tease apart the duration of reciprocal physical contact from gamete and accessory protein transfer.

Review of Harpactea ground-dwelling spiders (Araneae: Dysderidae) of Portugal

We revise the Portuguese spiders of the genus Harpactea Bristowe, 1939 (Araneae: Dysderidae). The following seven new species are described: H. adicensis sp. nov., H. crespoi sp. nov., H. dolanskyi sp. nov., H. henriquesi sp. nov., H. korenkoi sp. nov., H. krejcii sp. nov. and H. pekari sp. nov. In three species, H. algarvensis Ferrández, 1990, H. minoccii Ferrández, 1982 and H. tavirensis Wunderlich, 2020 the females are described for the first time. The Portuguese Harpactea spiders belong to two species groups, namely the hombergi group and the corticalis group (sensu Deeleman-Reinhold 1993). The majority of the Portuguese representatives of the corticalis group probably constitute a monophyletic group endemic for the Iberian peninsula, the minoccii subgroup Ferrández, 1990. The synapomorphy of this clade is the unique position of partners during copulation: the male grasps one of the female's chelicerae between his chelicerae. The males' chelicerae are morphologically adapted for this behaviour. Due to this unusual position during copulation the female genitalia are more distant from the male than in other Harpactea species. Therefore, the pedipalps are more elongated . Furthermore, the males of this subgroup possess larger basal part of male copulatory organ (the tegulum), where the ejaculate is stored before copulation. We observed that the males of this subgroup invest significantly more time into single copulation than the other representatives of the corticalis group with smaller tegulum. Therefore, enlargement of the tegulum might reflect different sperm competition strategy, in which males invest more ejaculate in each copulation. In Portugal, Harpactea spiders are frequently found under woody plants that produce slowly decomposing leaf litter, usually Quercus spp., or introduced Eucalyptus sp. These spiders require slightly humid substratum.

An observation of white whale (Delphinapterus leucas) mating behaviour in the wild

Herein we describe mating behaviour observed in a group of 20 white whales in Svalbard, Norway. A single female was the centre of attention during the 45-minute encounter, which was observed, photographed and videotaped at an ice edge in Storfjorden in June 2022. Several males surrounded the female and vigorously herded her from all sides. The female spy-hopped regularly, and sometimes her whole body was pushed out of the water by the other animals. High tail-lifts and tail-slapping were frequently performed by males, and erect penises were observed many times. Trumpet calls were also heard. At one point, three males with erect penises pressed themselves against the female simultaneously. The female was bleeding from wounds on her head and neck that were seemingly inflicted by bites from the other whales. She was also bleeding from her genital slit. A single copulation was seen at the surface, but it is likely that multiple males mated successfully with the female during the encounter. The event was more boisterous and violent than what has been described in captive white whales. We cannot assess how representative this observation is of typical reproductive behaviour for the species. But the encounter was remarkably similar to what has been described for several species of bottlenose dolphins that have male alliances that consort with individual females, restricting the female’s escape and keeping her accessible to mating only by members of the group.

Hidden impacts of conservation management on fertility of the critically endangered kākāpō.

Animal conservation often requires intensive management actions to improve reproductive output, yet any adverse effects of these may not be immediately apparent, particularly in threatened species with small populations and long lifespans. Hand-rearing is an example of a conservation management strategy which, while boosting populations, can cause long-term demographic and behavioural problems. It is used in the recovery of the critically endangered kākāpō (Strigops habroptilus), a flightless parrot endemic to New Zealand, to improve the slow population growth that is due to infrequent breeding, low fertility and low hatching success. We applied Bayesian mixed models to examine whether hand-rearing and other factors were associated with clutch fertility in kākāpō. We used projection predictive variable selection to compare the relative contributions to fertility from the parents' rearing environment, their age and previous copulation experience, the parental kinship, and the number of mates and copulations for each clutch. We also explored how the incidence of repeated copulations and multiple mates varied with kākāpō density. The rearing status of the clutch father and the number of mates and copulations of the clutch mother were the dominant factors in predicting fertility. Clutches were less likely to be fertile if the father was hand-reared compared to wild-reared, but there was no similar effect for mothers. Clutches produced by females copulating with different males were more likely to be fertile than those from repeated copulations with one male, which in turn had a higher probability of fertility than those from a single copulation. The likelihood of multiple copulations and mates increased with female:male adult sex ratio, perhaps as a result of mate guarding by females. Parental kinship, copulation experience and age all had negligible associations with clutch fertility. These results provide a rare assessment of factors affecting fertility in a wild threatened bird species, with implications for conservation management. The increased fertility due to multiple mates and copulations, combined with the evidence for mate guarding and previous results of kākāpō sperm morphology, suggests that an evolutionary mechanism exists to optimise fertility through sperm competition in kākāpō. The high frequency of clutches produced from single copulations in the contemporary population may therefore represent an unnatural state, perhaps due to too few females. This suggests that opportunity for sperm competition should be maximised by increasing population densities, optimising sex ratios, and using artificial insemination. The lower fertility of hand-reared males may result from behavioural defects due to lack of exposure to conspecifics at critical development stages, as seen in other taxa. This potential negative impact of hand-rearing must be balanced against the short-term benefits it provides.

EFFECTIVE TRAP FOR BLACK BEETLE (COLEOPTERA, TENEBRIONIDAE) AND OTHER INSECTS THAT CAUSE DAMAGE TO POULTRY

Darkling beetles Alphitobius diaperinus (Panzer, 1796) and A. laevigatus (Fabricius, 1781) (Coleoptera, Tenebrionidae) are the main pests of young chicks in poultry industry. A. diaperinus had already been recorded causing damage to the poultry industry in Crimea. Its larvae and adults attack chicks in the mass. Significant cannibalism is recorded for A. laevigatus in laboratory conditions. We suppose that complex of harmful insects in poultries will include species Ulomoides dermestoides (Chevrolat, 1878) which was introduced recently to Ukraine. Properties of these introduced harmful beetles, providing their survival and invulnerability to full liquidation in poultry houses, are firstly shown, namely: mass rearing of A. diaperinus due to their predaceous feeding on living birds, ability of females of U. dermestoides to oviposit eggs during 1,5 months after the single copulation, and prolongation of duration of the larval stages up to 96–110 days (usually 30 days long) due to a cannibalism only. The features of development and behavior of U. dermestoides are shown resembling these of A. diaperinus. New data on behavior, duration and peculiarities of life cycle were carried out during 2012–2019 under the laboratory conditions and received for the first time. The practical role of A. diaperinus, A. laevigatus and U. dermestoides is discussed. The forecast regarding the negative impact of U. dermestoides on the aviculture and poultry industry development in the Southern Ukraine is presented. Present contribution is beneficial not only for specialists in fundamental research, but also for practitioners, in particular, for the personnel of State Veterinary and Plant Health as well as the State Sanitary and Epidemiological Service. The structure and model of effective trap for collecting of larvae and adults of harmful darkling beetles at the presence of living birds in poultry houses is presented and recommended here for production and application in poultry industry.

Male nutritional status does not impact the reproductive potential of female Cnaphalocrocis medinalis moths under conditions of nutrient shortage.

In addition to sperm, some accessory substances transferred to females during copulation act as nuptial gifts by passing on valuable nutrients in many insect species. The nutritional status of the males can thus have a great effect on the mating behavior, fecundity and even the longevity of females. However, little is known about the effect of male nutritional status on the female reproductive traits in migratory insect species, particularly when females experience nutrient shortage and have to choose between reproduction and migration. Here, Cnaphalocrocis medinalis, a migratory rice pest in Asia, was studied to explore this issue. Our results showed that in male moths fed with honey solution, their gonads had higher energy content than gonads of starved males, resulting in increased energy content of the bursa copulatrix of females after mating with fed males. Such females showed increased mating frequency, fecundity and longevity compared to females mating with starved males, indicating that male moths deliver nuptial gifts to females and improve their reproductive performance. However, when females were starved, only about 45% mated, with just a single copulation, regardless of male nutritional status. Starved females showed lower fecundity, and a longer pre-oviposition period (indicating a greater propensity to migrate), compared to fed females. However, copulation still significantly extended their longevity. These results suggest that starved females invest in migration to escape deteriorating habitats, rather than investing the nuptial gift to increased fecundity. Our results further our understanding of the reproductive adaptability of migratory insects under conditions of food stress.

From sperm plug formation to ovulation: morphological and ultrastructural modifications in the seminal receptacle of the blue crab Callinectes danae

In species of Callinectes a sperm plug is formed in the seminal receptacle (SR) during the sperm transfer, which hinders the insemination by other males. In this study, we evaluated the morphology, histochemistry, and ultrastructure of the seminal receptacle of juvenile and adult females of Callinectes danae along the different stages of ovarian development. We described the histochemical changes that take place inside the SR, from the sperm plug formation until fertilization. We also quantified the amount of seminal fluid received by the females during copulation. Callinectes danae has a dorsal-type SR, which is common in Portunidae. The sperm plug is formed by the orderly deposition of seminal fluid of the anterior and median vas deferens, which in turn influences the secretion of the posterior region and produces chemically distinct strata. The dissolution of the sperm plug inside the SR occurs in synchrony with the spermatophore dehiscence and ovarian development. A holocrine secretion from dense layer cells dissolves the sperm plug. These cells have electron-dense and electron-lucid vesicles. The plug is completely dissolved when the females are at the intermediary ovarian development. At this stage, the SR dense layer folds and forms the lateral chamber. The dorsal oviduct, continuous to the dense layer, moves to a ventral-lateral position opposed to the vagina. During ovulation, the oocytes of mature females pass through the oviduct and the fertilization occurs almost above the vagina. The male ejaculates all its seminal fluid in a single copulation, leading to a total sperm depletion. Therefore, the whole sperm plug is produced by the males, through the combined secretions from three vas deferens regions. We show, for the first time, that the secretions from the anterior vas deferens also participate in the sperm plug formation. The female dissolves the plug, which is a crucial morphological change of the SR during the processes of fertilization and egg release. In species of Callinectes there was a clear co-evolution between the male and female reproductive systems: the male invests all its reproductive potential to build the sperm plug, and the female, to dissolve it.

Mating-regulated atrial proteases control reinsemination rates in Anopheles gambiae females

Anopheles gambiae mosquitoes are the most important vectors of human malaria. The reproductive success of these mosquitoes relies on a single copulation event after which the majority of females become permanently refractory to further mating. This refractory behavior is at least partially mediated by the male-synthetized steroid hormone 20-hydroxyecdysone (20E), which is packaged together with other seminal secretions into a gelatinous mating plug and transferred to the female atrium during mating. In this study, we show that two 20E-regulated chymotrypsin-like serine proteases specifically expressed in the reproductive tract of An. gambiae females play an important role in modulating the female susceptibility to mating. Silencing these proteases by RNA interference impairs correct plug processing and slows down the release of the steroid hormone 20E from the mating plug. In turn, depleting one of these proteases, the Mating Regulated Atrial Protease 1 (MatRAP1), reduces female refractoriness to further copulation, so that a significant proportion of females mate again. Microscopy analysis reveals that MatRAP1 is localized on a previously undetected peritrophic matrix-like structure surrounding the mating plug. These data provide novel insight into the molecular mechanisms shaping the post-mating biology of these important malaria vectors.

Dysfunction of dimorphic sperm impairs male fertility in the silkworm

Sperm, which have a vital role in sexual reproduction in the animal kingdom, can display heteromorphism in some species. The regulation of sperm dichotomy remains a longstanding puzzle even though the phenomenon has been widely documented for over a century. Here we use Bombyx mori as a model to study a form of sperm dimorphism (eupyrene and apyrene sperm), which is nearly universal among Lepidoptera. We demonstrate that B. mori Sex-lethal (BmSxl) is crucial for apyrene sperm development, and that B. mori poly(A)-specific ribonuclease-like domain-containing 1 (BmPnldc1) is required for eupyrene sperm development. BmSXL is distributed in the nuclei and cytoplasm of somatic cyst cells in a mesh-like pattern and in the cytoplasm of germ cells enclosed in spermatocysts and sperm bundles. Cytological analyses of dimorphic sperm in BmSxl mutants (∆BmSxl) showed deficient apyrene sperm with abnormal nuclei, as well as loss of motility associated with malformed mitochondrial derivatives. We define the crucial function of apyrene sperm in the process of fertilization as assisting the migration of eupyrene spermatozoa from bursa copulatrix to spermatheca. By contrast, BmPnldc1 deficiency (∆BmPnldc1) caused eupyrene sperm abnormalities and impaired the release of eupyrene sperm bundles during spermiation. Although apyrene or eupyrene sperm defects impaired fertility of the mutated males, double copulation of a wild-type female with ∆BmSxl and ∆BmPnldc1 males could rescue the sterility phenotypes induced by single copulation with either gene-deficient male. Our findings demonstrate the crucial functions of BmSxl and BmPnldc1 in the development of sperm dimorphism and the indispensable roles of nonfertile apyrene sperm in fertilization.

Does sexual cannibalism secure genetic benefits of polyandry in a size-dimorphic spider?

Females mate multiply despite numerous costs. It is well established that polyandry can result in sexual conflict, favoring male adaptations that prevent sperm competition often to the disadvantage of the female. Such adaptations are extreme in spiders with one-shot genitalia of which parts break off and act as mating plugs, rendering them dysfunctional. In the spider Argiope bruennichi, mating plugs effectively prevent further males from inseminating and males that inseminate and plug both genital openings of a female secure exclusive paternity. However, females frequently prevent monopolization by attacking and cannibalizing males during their first copulation, leaving their second spermatheca free for another male. Here, we test whether the high frequency of sexual cannibalism evolved as a female adaptation to resist monopolization and secure indirect benefits of polyandry. To standardize conditions, we double-mated females either with the same or two different males and prevented male consumption. Using a split-brood design, we raised offspring to maturity under poor and rich food conditions and measured their survival, duration of juvenile phase, and adult body mass. Under low food, daughters of polyandrous mothers matured later but slightly heavier than daughters of monandrous females. Since the adaptive value of this combination is unclear, these findings lend no conclusive support to our hypothesis. We discuss the stereotypic nature of the female attack in the context of antagonistic co-evolution considering previous studies that found modest direct benefits of cannibalism as well as a potential for non-additive benefits.Significance statementSexual conflict is extreme in spiders where sexual cannibalism impairs male mating rates. Males of the spider Argiope bruennichi possess one-shot genitalia which they break off to plug female genital openings. They gain exclusive paternity with a female if two copulations are achieved and both genital openings plugged. Females, however, stereotypically attack every male at the onset of copulation, limiting most males to single copulation but retaining the option to secure potential benefits of polyandry. Previous studies revealed weak direct and non-additive indirect benefits of multiple mating. In this study, we tested for the presence of additive genetic benefits but again found only inconclusive evidence for adaptive differences in offspring quality between monandrous and polyandrous females. All results combined, we here speculate that the stereotypic female attack might be a ghost of a past antagonistic co-evolution.

Males of a sexually cannibalistic spider chemically assess relative female quality

BackgroundMate choice is a taxonomically wide-spread phenomenon, mostly exerted by females although male mate choice occurs as well. While costs and benefits of choosiness have been well studied, the underlying mechanisms are largely unclear. Different models exist, namely best-of-n or threshold rules, based on sequential or simultaneous sampling, which differ in the required cognitive demands. We applied an experimental approach to shed light on the underlying mechanisms of male mate choice in the sexually cannibalistic spider Argiope bruennichi. Males are limited to two copulations and preferentially monopolise large females, while they may leave smaller females after a single copulation and resume mate search. Here, we utilised significant size-differences between females from Northern and Southern populations and presented males with three different-sized females that were matched for origin: all three females originated either from the same Northern European population as the males or from Southern populations where the smallest female was about the same size as the largest Northern female. This allowed testing the hypothesis that males base their mating tactic on a fixed local size threshold. We predicted Northern males to be choosy among Northern females, but to accept all Southern females since they would all be above that threshold.ResultsMales copulated with the first female they encountered, which was independent of her body size. Regardless of the females’ origins, males chose a monogynous tactic with the largest female in the trio, while they left the smallest female after one copulation. The same pattern applied to Southern females even though the smallest females in the trio were of a similar size as monopolised Northern females. Since males have poor eyesight and did not actively sample all females, they likely have gained information about relative size differences between females based on volatile chemical cues only.ConclusionsOur findings suggest that male A. bruennichi can assess relative differences in mate quality and adjust their mating tactic to the prevailing conditions (Northern vs. Southern). We reject the presence of a locally-adapted fixed threshold and argue that our results are best explained by an adjustable threshold that was raised under Southern conditions.

Fitness advantages of the biased use of paired laterally symmetrical penises in an insect.

The evolution of laterality, that is the biased use of laterally paired, morphologically symmetrical organs, has attracted the interest of researchers from a variety of disciplines. It is, however, difficult to quantify the fitness benefits of laterality because many organs, such as human hands, possess multimodal functions. Males of the earwig Labidura riparia (Insecta: Dermaptera: Labiduridae) have morphologically similar laterally paired penises, only one of which is used for inseminating the female during a single copulation bout, and thus provide a rare opportunity to address how selection pressure may shape the evolution of population-level laterality. Our population studies revealed that in 10 populations, located at 2.23-43.3° north, the right penis is predominantly used for copulating (88.6%). A damaged penis was found in 23% of rare left-handers, suggesting that the left penis can function as a spare when the right one is damaged. By pairing L.riparia females with surgically manipulated males, we found that males forced to use the right penis outperformed left-handed males in copulation (the probability of establishing genital coupling during the 1-hr observation period: odds ratio [OR] of 3.50) and insemination (probability of transferring a detectable amount of sperm: OR of 2.94). This right-handed advantage may be due to the coiled morphology of the sperm storage organ with a right-facing opening. Thus, female genital morphology may play a significant role in the evolution of handedness and may have acted as a driving force to reduce penis number in related taxa.

Mate availability does not influence mating strategies in males of the sexually cannibalistic spider Argiope bruennichi.

BackgroundSexual selection theory predicts that male investment in a current female should be a function of female density and male competition. While many studies have focused on male competition, the impact of female density on male mating investment has been widely neglected. Here, we aimed to close this gap and tested effects of mate density on male mating decisions in the orb-web spider Argiope bruennichi. Males of this species mutilate their genitalia during copulation, which reduces sperm competition and limits their mating rate to a maximum of two females (bigyny). The mating rate is frequently further reduced by female aggression and cannibalization. Males can reduce the risk of cannibalism if they jump off the female in time, but will then transfer fewer sperm. An alternative solution of this trade-off is to copulate longer, commit self-sacrifice and secure higher minimal paternity. The self-sacrificial strategy may be adaptive if prospective mating chances are uncertain. In A. bruennichi, this uncertainty may arise from quick changes in population dynamics. Therefore, we expected that males would immediately respond to information about low or high mate availability and opt for self-sacrifice after a single copulation under low mate availability. If male survival depends on information about prospective mating chances, we further predicted that under high mate availability, we would find a higher rate of males that leave the first mating partner to follow a bigynous mating strategy.MethodWe used naïve males and compared their mating decisions among two treatments that differed in the number of signalling females. In the high mate availability treatment, males perceived pheromone signals from four adult, virgin females, while in the low mate availability treatment only one of four females was adult and virgin and the other three were penultimate and unreceptive.ResultsMales took more time to start mate searching if mate availability was low. However, a self-sacrificial strategy was not more likely under low mate availability. We found no effects of treatment on the duration of copulation, the probability to survive the first copulation or the probability of bigyny. Interestingly, survival chances depended on male size and were higher in small males.DiscussionOur results do not support the hypothesis that mate density variation affects male mating investment, although they clearly perceived mate density, which they presumably assessed by pheromone quantity. One reason for the absence of male adjustments to mating tactics could be that adaptations to survive female attacks veil adaptations that facilitate mating decisions.

Characterizing estrus by trans-abdominal ultrasounds, fecal estrone-3-glucuronide, and vaginal cytology in the Steller sea lion (Eumetopias jubatus)

The ability to monitor the estrus cycle in wild and captive marine species is important for identifying reproductive failures, ensuring a successful breeding program, and monitoring animal welfare. Minimally invasive sampling methods to monitor estrus in captive populations have been developed, but results suggest these tools can be species-specific in their precision and accuracy. Therefore, the minimally invasive sampling methods of trans-abdominal ultrasounds, a fecal steroid analysis (estrone-3-glucuronide, E1G), and vaginal cytology, were evaluated for their efficacy to characterize and monitor estrus in a captive breeding population of Steller sea lions (Eumetopias jubatus). Three adult females were sampled over five breeding seasons, resulting in six estrus profiles characterized by trans-abdominal ultrasounds, five by fecal E1G, and four by vaginal cytology. Animals were trained to allow trans-abdominal ultrasounds, fecal samples, and vaginal swabs to be collected approximately daily. Of the 76 trans-abdominal ultrasound sessions attempted, 8 successfully visualized both ovaries. From these scans, the chronology of ovarian changes during proestrus and estrus was estimated. The time from the detection of developing follicles to the identification of a dominate follicle occurred in 2–5 days and a corpus hemorrhagicum formed approximately 4 days later. However, because visualization of the ovaries was prevented by the gastrointestinal system in 88% of scans, this tool was overall unreliable for monitoring changes associated with estrus. To detect fine scale physiological changes associated with estrus, we analyzed changes in fecal E1G (n = 62) and vaginal cytology (n = 157) 15 days before and after each female's single copulation event (Day = 0). Changes in fecal E1G had the highest accuracy at detecting Day = 0. Fecal E1G increased leading up to estrus, peaked at Day = 0, and then declined. Although we did observe the characteristic increase in superficial cells associated with impending estrus, the type of cell which peaked closest to Day = 0 was intermediate. The uncertainty around the peak in intermediate cells, indicating estrus, was greater than the uncertainty associated with detecting estrus from fecal E1G. Collectively, these results suggest that changes in fecal E1G and vaginal cytology are viable tools to detect estrus in Steller sea lions, but require daily sampling to detect gradual changes, limiting their applicability to studies of wild populations.

Polyandry enhances offspring viability with survival costs to mothers only when mating exclusively with virgin males in Drosophila melanogaster.

A prominent hypothesis for polyandry says that male–male competitive drivers induce males to coerce already‐mated females to copulate, suggesting that females are more likely to be harassed in the presence of multiple males. This early sociobiological idea of male competitive drive seemed to explain why sperm‐storing females mate multiply. Here, we describe an experiment eliminating all opportunities for male–male behavioral competition, while varying females’ opportunities to mate or not with the same male many times, or with many other males only one time each. We limited each female subject's exposure to no more than one male per day over her entire lifespan starting at the age at which copulations usually commence. We tested a priori predictions about relative lifespan and daily components of RS of female Drosophila melanogaster in experimental social situations producing lifelong virgins, once‐mated females, lifelong monogamous, and lifelong polyandrous females, using a matched‐treatments design. Results included that (1) a single copulation enhanced female survival compared to survival of lifelong virgins, (2) multiple copulations enhanced the number of offspring for both monogamous and polyandrous females, (3) compared to females in lifelong monogamy, polyandrous females paired daily with a novel, age‐matched experienced male produced offspring of enhanced viability, and (4) female survival was unchallenged when monogamous and polyandrous females could re‐mate with age‐ and experienced‐matched males. (5) Polyandrous females daily paired with novel virgin males had significantly reduced lifespans compared to polyandrous females with novel, age‐matched, and experienced males. (6) Polyandrous mating enhanced offspring viability and thereby weakened support for the random mating hypothesis for female multiple mating. Analyzes of nonequivalence of variances revealed opportunities for within‐sex selection among females. Results support the idea that females able to avoid constraints on their behavior from simultaneous exposure to multiple males can affect both RS and survival of females and offspring.

Delayed sexual maturity in males of Vespa velutina.

Vespa velutina var nigrithorax (Lepelletier, 1835) is an invasive predator of bees accidentally introduced in France in 2004, and it is having a serious impact on apiculture and ecosystems. Studying the reproduction of an invasive species is key to assess its population dynamic. This study explores the sexual maturation of V. velutina males and the evolution of their fertility. The main studied parameters were physiologic (spermiogenesis, spermatogenesis) and anatomic (testes size and structure, head width). Two populations of males were described based on their emergence period: early males in early summer or classic males in autumn. Each testis has an average of 108 testicular follicles. Spermatogenesis is synchronous, with only 1 sperm production wave, and completed, on average, at 10.3 d after emergence with the degeneration of the testes. The sperm counts in seminal vesicles of mature males are 3 × 106 in October/November and 0.8 × 106 in June. In comparison, females store 0.1 × 106 sperm in their spermathecae. The early males emerged from colonies made by fertilized queens. The reproductive potential of these early males seemed limited, and their function in the colony is discussed. The sperm stock evolution in autumn males suggests the occurrence of a reproductive pattern of male competition for the access to females and a single copulation per male. The synchronicity of male and foundress emergences and sexual maturation is of primary importance for the mating success and the future colony development.

Impact of cryptic female choice on insemination success: Larger sized and longer copulating male squid ejaculate more, but females influence insemination success by removing spermatangia.

In polyandrous mating systems, sperm competition and cryptic female choice (CFC) are well recognized as postcopulatory evolutionary forces. However, it remains challenging to separate CFC from sperm competition and to estimate how much CFC influences insemination success because those processes usually occur inside the female's body. The Japanese pygmy squid, Idiosepius paradoxus, is an ideal species in which to separate CFC from sperm competition because sperm transfer by the male and sperm displacement by the female can be observed directly at an external location on the female's body. Here, we counted the number of spermatangia transferred to, removed from, and remaining on the female body during single copulation episodes. We measured behavioral and morphological characteristics of the male, such as duration of copulation and body size. Although males with larger body size and longer copulation time were capable of transferring larger amounts of sperm, females preferentially eliminated sperm from males with larger body size and shorter copulation time by spermatangia removal; thus, CFC could attenuate sperm precedence by larger males, whereas it reinforces sperm precedence by males with longer copulation time. Genetic paternity analysis revealed that fertilisation success for each male was correlated with remaining sperm volume that is adjusted by females after copulation.

Diet affects ejaculate traits in a lizard with condition-dependent fertilization success

Sexually selected traits are often driven to costly extremes by persistent directional selection. Energy acquisition and allocation can therefore influence variation in traits subject to both precopulatory and postcopulatory sexual selection, though the later have received much less attention. We tested the condition dependence of sperm morphology, sperm count, and fertilization success in a promiscuous lizard (Anolis sagrei) by 1) collecting sperm samples from wild males that varied naturally in body condition, 2) experimentally altering the body condition of captive males through dietary restriction, and 3) analyzing genetic paternity data from competitive mating trials between captive males that differed in body condition. In both wild and captive males, the length of the sperm midpiece decreased with body condition. Experimental food restriction decreased sperm production, decreased length of the sperm head, increased length of the sperm midpiece, and increased variance in sperm morphology within individuals. When restricted to a single copulation, males on high-intake diets exhibited a slight but nonsignificant fertilization advantage. Reanalysis of a previous experiment in which high- and low-condition males were sequentially allowed to copulate ad libitum for 1 week revealed a significant fertilization bias in favor of high-condition males. When controlling for mean treatment effects on the proportion of offspring sired and on sperm phenotypes, multiple regression revealed negative correlations between fertilization success and sperm head length, midpiece length, and sperm count. Collectively, our results suggest that condition-dependent fertilization success in A. sagrei may be partially mediated by underlying condition dependence of sperm morphology and sperm count.

Promiscuity and elongated sperm storage organs work cooperatively as a cryptic female choice mechanism in an earwig

Female genitalia often show complex morphologies that cannot be explained by sperm reception and storage functions. However, our understanding of the forces underlying genital exaggeration in females is limited. Female earwigs, Euborellia plebeja , are promiscuous and their highly elongated sperm storage organs allow only partial removal and displacement of stored sperm by shorter male genitalia, resulting in only a 20% gain in paternity per copulation with a sperm-saturated female. This study examined the significance and optimality of restricted sperm displacement for females. A staged mating experiment with a paternity success analysis revealed that large males dominated male–male competition for burrows housing females, resulting in repeated copulations with the same female. Despite the low paternity gain per copulation, such repeated copulations resulted in a significant increase in paternity for larger males with higher resource-holding potential. A numerical simulation based on the relationship between male body size and copulation frequency showed that restricted sperm displacement (about 20% per copulation) is optimal for promiscuous females to accumulate sperm effectively from larger males. Because male body size is heritable in this species, females were estimated to benefit from a 1.4% increase in their sons' mating success. This genetic benefit disappeared when only a single copulation per male–female encounter was assumed. Since no measurable costs of mating for females have been detected in this species, the combination of promiscuity and restricted sperm displacement is best explained by the cryptic female choice hypothesis.