Sexual coercion does not exclude luring behavior in the climbing camel-spider <Emphasis Type="Italic">Oltacola chacoensis</Emphasis> (Arachnida,Solifugae, Ammotrechidae)

Sexual coercion in the form of forced copulation has been used as a typical example to illustrate the conflict of interests between females and males. Among arthropods, forced copulation has been reported for some groups of insects and crustaceans, but not for arachnids. In the present work, we analyse and describe the behavioral patterns of mating behavior of the climbing camel-spider, Oltacola chacoensis, relating it to relevant morphological features, In this species, the male forcefully clasps the female’s genital region with his chelicerae and locks her fourth pair of legs with his pedipalps. In some cases, the cuticle of the female’s abdomen was damaged by this cheliceral clasping. In contrast to other camel-spiders, the female O. chacoensis never remained motionless during mating, but continuously shook her body, opening her chelicerae notably towards the male. Despite this coercive context, males performed copulatory courtship (tapping with pedipalps) and females showed an apparent cooperative behavior (they remained still during a short period of the sperm transfer phase). These results strengthen the idea that sexual coercion (in the form of forced copulation) and luring behavior (in the form of copulatory courtship) are not two mutually-exclusive male’s strategies during a single copulation.     

Female behaviour and the interaction of male and female genital traits mediate sperm transfer during mating

Natural selection and post‐copulatory sexual selection, including sexual conflict, contribute to genital diversification. Fundamental first steps in understanding how these processes shape the evolution of specific genital traits are to determine their function experimentally and to understand the interactions between female and male genitalia during copulation. Our experimental manipulations of male and female genitalia in red‐sided garter snakes (Thamnophis sirtalis parietalis) reveal that copulation duration and copulatory plug deposition, as well as total and oviductal/vaginal sperm counts, are influenced by the interaction between male and female genital traits and female behaviour during copulation. By mating females with anesthetized cloacae to males with spine‐ablated hemipenes using a fully factorial design, we identified significant female–male copulatory trait interactions and found that females prevent sperm from entering their oviducts by contracting their vaginal pouch. Furthermore, these muscular contractions limit copulatory plug size, whereas the basal spine of the male hemipene aids in sperm and plug transfer. Our results are consistent with a role of sexual conflict in mating interactions and highlight the evolutionary importance of female resistance to reproductive outcomes.     

Sperm dumping in a haplogyne spider

The present study shows that females of Silhouettella loricatula (Arachnida: Araneae: Oonopidae) manage to process sperm in an unusual and previously unknown way. The male ejaculate consisting of spermatozoa and globular secretion is enclosed in a secretory sac. This may avoid the mixing of sperm from different males and at least severely limit sperm competition. The process of sperm enclosure occurs within the female's sperm storage site (receptaculum) as the ejaculate is not surrounded by a sac inside the male's sperm-transferring organs (palpal bulbs). The secretion forming the sac is produced by glands adjoining the receptaculum. The possibility that globular secretions in the male palpal bulbs partly contribute to the sac cannot be ruled out completely. It is suggested that in S. loricatula , the main function of sperm enclosure in a sac is enabling females to dump the ejaculate of a male. The present study represents the first report on sperm dumping in the family Oonopidae. During five first and three second copulations in the laboratory, the dumping of a sac was observed. One dumped sac was sectioned and contained spermatozoa. Two couples were flash-fixed with liquid nitrogen early during copulation, which revealed the mechanism of the sac dumping. By muscle contractions, the receptaculum is bent backwards and the sac moved into the genital opening. The actual sac dumping occurs most probably in cooperation with the male, which moves his pedipalps rhythmically during the entire copulation. Extensions and furrows on the emboli suggest that they may additionally be used as copulatory courtship devices. The enclosure of sperm from the current male in secretion takes place during or immediately following copulation as all mated females sacrificed after copulation had a new sac containing spermatozoa in the receptaculum. Dumping sperm of a previous male during the next copulation may allow females to bias sperm precedence.     

Determinants of sperm transfer in the scorpionfly Panorpa cognate: male variation, female condition and copulation duration

Recent studies suggest that sperm production and transfer may have significant costs to males. Male sperm investment into a current copulation may therefore influence resources available for future matings, which selects for male strategic mating investment. In addition, females may also benefit from actively or passively altering the number of sperm transferred by males. In the scorpionfly Panorpa cognata, the number of sperm transferred during copulation depended on copulation duration and males in good condition (residual weight) copulated longer and also transferred more sperm. Moreover, sperm transferred and stored per unit time was higher in copulations with females in good condition than in copulations with females in poor condition. Males varied greatly and consistently in their sperm transfer rate, indicative of costs associated with this trait. The duration of the pairing prelude also varied between males and correlated negatively with the male's sperm transfer rate, but no other male character correlated significantly with male sperm transfer rate. The results are consistent with strategic mating effort but sperm transfer could also be facilitated by the physical size of females and/or females in good condition may be more cooperative during sperm transfer.     

Mating duration and sperm precedence in the spider <Emphasis Type="Italic">Linyphia triangularis</Emphasis>

In many animal species, mating behaviour is highly ritualised, which may allow us to relate some of its consequences, e.g. male paternity and female receptivity, to the progression of phases in the mating sequence; at the same time, ritualisation raises the question of to what extent the partners, especially the males, are able to influence the outcome of mating for their own benefit. We studied the linyphiid spider Linyphia triangularis in which mating follows a strict sequence during which the male inducts two droplets of sperm and transfers them to the female. We performed sperm competition experiments (sterile-male technique) including four treatments, in which the copulation of the first male was interrupted at prescribed phases of the mating sequence, while the second male was allowed a complete mating. Second males spent a shorter time than first males on the behaviours prior to sperm transfer, but the amount of sperm (2 droplets) and the time spent in sperm transfer were independent of the females’ mating status. The proportion of females accepting the second male depended on the mating duration of the first male, i.e. whether the first male had transferred one or two sperm droplets. After a complete first mating, most females accepted no further males. A last-male sperm precedence was apparent if only half of the first sperm droplet had been transferred by the first male, but this switched to a first male precedence if one full sperm droplet had been transferred. Thus, even in the face of sperm competition, it is sufficient for the first male to transfer one sperm droplet. The second sperm droplet and the extended copulatory courtship associated with its transfer may serve to induce a lack of receptivity in the female, but the males seem unable to enhance their reproductive success through variable copulatory tactics.     

Securing paternity: mating plugs in the dwarf spider Oedothorax retusus (Araneae: Erigoninae)

One of the various male strategies to prevent or impede female remating is the production of a mating plug that covers the female genital opening or remains inside of the female genital tract after mating. Such structures have been described for many species in many animal taxa; however, in most cases, we know little or nothing about their specific adaptive value. Our investigations demonstrate that females of the dwarf spider species Oedothorax retusus (Westring, 1851) (Linyphiidae, Erigoninae) exhibit a substance on one or both of her paired genital openings only after copulation. We performed double-mating trials and forced the second male to mate into the previously used or unused spermathecal duct of the female by amputating one of his paired male gonopods (pedipalps). Furthermore, to investigate whether the duration of the first mating has an effect on the size and efficiency of the mating plug, we interrupted first matings after either 1 or 3 min, categorized plug size and recorded mating behaviour of subsequent males. The amount of secretion transferred was larger in long compared to short copulations. A long first copulation successfully prevented subsequent males from mating into the used ducts, whereas mating success after short first matings was similar to matings into unused copulatory ducts of the females. The present study demonstrates that a male O. retusus can prevent a rival from transferring sperm into the same spermatheca by applying a mating plug, but only if he mates for long enough.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 96 , 574–583.     

Patterns of Sperm Transfer in the Golden Orb‐Weaver Nephila edulis

Sperm competition studies typically identify copulation duration as an important predictor of paternity as it may determine the quantity of sperm transferred and thus paternity success. This study explores the relationship between copulation duration, male body size, male age and sperm transfer in the golden orb‐weaving spider, Nephila edulis. Paternity in this species is strongly associated with the relative frequency and duration of copulation, which is also influenced by male size. We determined the number of sperm transferred during copulation, by performing sperm counts in both the male copulatory organs (palps) and female sperm receptacle (spermatheca) of recently mated pairs. The total number of sperm recorded (the sum in the male palps and female spermathecae) was greater for younger males than older males, but did not vary with male body size. In general, younger males transferred more sperm and a greater proportion of their sperm supplies than older males and, among these younger males, larger individuals transferred more sperm. However, there were no significant size effects for older males. More sperm was transferred with longer copulations, but in contrast with previous studies, we found that larger males copulated for longer. The rate of sperm transferred was negatively correlated with the duration of copulation, suggesting that the variation in copulation duration in N. edulis may represent strategic investment by males to alter patterns of paternity, in addition to transferring additional sperm.     

Female wolf spiders exert cryptic control drastically reducing ejaculate size

Sexes' roles in post‐copulatory processes have important effects on individual fitness and are promising to study in species showing complex mating behaviours. In the spider Schizocosa malitiosa, males perform two different copulatory patterns, pattern 1 includes 80% of total pedipalp insertions and pattern 2 includes 20%. Both patterns produce similar number of offspring, but pattern 1 induces higher female reluctance to remating than pattern 2. We hypothesised that the complex copulatory patterns are linked to post‐copulatory sexual selection, affecting males' sperm transfer and the resulting sperm storage by females. First, we examined amounts of sperm in males and live females from uninterrupted (pattern 1 + 2) and interrupted matings (pattern 1, pattern 2). Second, in order to disentangle male and female actions, we induced males to mate with dead females and examined amounts of sperm. Males transfer in total 71% of the sperm available in their pedipalps, being higher but not significant in pattern 1 than in pattern 2. Females drastically reduced the amount of sperm stored in their spermathecae and such control is stronger in pattern 1 compared to pattern 2 matings. We propose that cryptic female control is a main factor driving males to strengthen sperm transfer. Active female reduction in ejaculate most probably diminished her reluctance to remate.     

Sperm storage and copulation duration in a sexually cannibalistic spider

Female St Andrew’s Cross spiders control copulation duration by timing sexual cannibalism and may thereby control paternity if cannibalism affects sperm transfer. We have investigated the effect of copulation duration on sperm transfer and documented sperm storage patterns when we experimentally reduced the ability of females to attack and cannibalise the male. Virgin males and females were paired and randomly allocated either to a control treatment, where females were allowed to attack and cannibalise the male during copulation, or to an experimental treatment, where females were unable to cannibalise the male. The latter was achieved by placing a paintbrush against her chelicerae during copulation. Our experimental manipulation did not affect copulation duration or sperm storage. However, the number of sperm stored by the female increased with copulation duration only if the male was cannibalised, suggesting that cannibalism increases relative paternity not only through prolonged copulation duration following a fair raffle model but also through the cannibalism act itself. Future studies should explore whether cannibalised males ejaculate more sperm or whether females selectively store the sperm of cannibalised males.     

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

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

Copulatory mechanism in a sexually cannibalistic spider with genital mutilation (Araneae: Araneidae: Argiope bruennichi)

Genitalia are among the fastest evolving morphological traits as evidenced by their common function as diagnostic traits in species identification. Even though the main function of genitalia is the successful transfer of spermatozoa, the presence of diverse structures that are obviously not necessary for this suggests that genitalia are a target of sexual selection. The male genitalia of many spider species are extremely complex and equipped with numerous sclerites, plates and spines whose functions are largely unknown. Selection on male genitalia may be particularly strong in sexually cannibalistic spiders, where mating success of males is restricted to a single female. We investigated the copulatory mechanism of the sexually cannibalistic orb weaving spider Argiope bruennichi by shock freezing mating pairs and revealed a complicated interaction between the appendices and sclerites that make up the male gonopods (paired pedipalps). The plate that covers the female genital opening (scape) is secured between two appendices of the male genital bulb, while three sclerites that bear the sperm duct are unfolded and extended into the female copulatory opening. During copulation, females attack and cannibalise the male and males mutilate their genitalia in about 80% of cases. Our study demonstrates that (i) genital coupling is largely accomplished on the external part of the female genitalia, (ii) that the mechanism requires an interaction between several non-sperm-transferring structures and (iii) that there are two predetermined breaking points in the male genitalia. Further comparative work on the genus Argiope will test if the copulatory mechanism with genital mutilation indeed is an adaptation to sexual cannibalism or if cannibalism is a female counter adaptation to male monopolisation through genital plugging.     

Sperm storage mediated by cryptic female choice for nuptial gifts

Polyandrous females are expected to discriminate among males through postcopulatory cryptic mate choice. Yet, there is surprisingly little unequivocal evidence for female-mediated cryptic sperm choice. In species in which nuptial gifts facilitate mating, females may gain indirect benefits through preferential storage of sperm from gift-giving males if the gift signals male quality. We tested this hypothesis in the spider Pisaura mirabilis by quantifying the number of sperm stored in response to copulation with males with or without a nuptial gift, while experimentally controlling copulation duration. We further assessed the effect of gift presence and copulation duration on egg-hatching success in matings with uninterrupted copulations with gift-giving males. We show that females mated to gift-giving males stored more sperm and experienced 17% higher egg-hatching success, compared with those mated to no-gift males, despite matched copulation durations. Uninterrupted copulations resulted in both increased sperm storage and egg-hatching success. Our study confirms the prediction that the nuptial gift as a male signal is under positive sexual selection by females through cryptic sperm storage. In addition, the gift facilitates longer copulations and increased sperm transfer providing two different types of advantage to gift-giving in males.     

Male copulatory sensory stimulation induces female ejection of rival sperm in a damselfly

Male damselflies possess very specialized genitalia. Females mate multiply and store sperm in two sperm storage organs, the bursa copulatrix and the spermatheca. During copulation, males physically remove the sperm stored in these organs using their genitalia. I document a novel mechanism by which males gain access to the spermatheca in Calopteryx haemorrhoidalis asturica. The mechanism is based on male stimulation of the female sensory system that controls egg fertilization and laying. During copulation, the aedeagus (a male genitalic structure indirectly involved in sperm transfer) distorts the cuticular plates in the female genital tract that bear mechanoreceptive sensilla. This stimulation results in sperm ejection from the spermatheca. Aedeagus width is positively correlated with the amount of sperm ejected. I propose that males have exploited a pre-existing female sensory bias to gain access to otherwise physically unreachable sperm. These results shed light on the issue of the origin of female preferences in current models of sexual selection and on the evolution of genitalia via sexual selection. It is postulated that females might use this process as a form of post-copulatory sexual selection on the basis of males'' genitalia.     

Sperm transfer and paternity in the scorpionfly <Emphasis Type="Italic">Panorpa cognata</Emphasis>: large variance in traits favoured by post-copulatory episodes of sexual selection

Post-copulatory episodes of sexual selection can be a powerful selective force influencing the reproductive success of males. In order to understand variation in male fertilisation success, we first need to consider the pattern of sperm utilisation by females following matings with more than one male. Second, we need to study those traits responsible for male success in sperm competition. Here we study both male sperm transfer characteristics as well as offspring paternity of females mated to two males in the scorpionfly Panorpa cognata. By repeatedly mating males to virgin females and interrupting copulation at defined time points, we found for all males that sperm transfer set off after approximately 40 min. During the remaining copulation, sperm transfer of individual males was continuous and with constant rate. Yet the rate of sperm transfer differed between individual males from about one sperm per minute to more than eight sperm per minute for the most successful males. In addition, we measured the fertilisation success in sperm competition of males with known sperm transfer capability. The relative number of sperm transferred by males during copulation, estimated from copulation duration and the males’ individual sperm transfer rate, explained a large proportion of variation in offspring paternity. The mode of sperm competition in this species, thus, conforms largely to a fair raffle following complete mixing of sperm prior to fertilisation. Hence, male differences in both the ability to copulate for long and of rapid sperm transfer will translate directly into differences in reproductive success.     

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

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

Adjustment of copula duration and ejaculate size according to the risk of sperm competition in the golden egg bug (Phyllomorpha laciniata)

Several hypotheses have been proposed to explain the adaptivesignificance of prolonged copulations in insects, which includemate guarding and sperm loading functions. We have exploredthe adaptive significance of the prolonged copulations in thegolden egg bug (copulations up to 50 h) and the effect of anincreased risk of sperm competition on ejaculate investment.Our data support predictions derived from sperm competitiontheory, which posits that males are expected to increase ejaculateexpenditure in response to an increased risk of sperm competition.Results show a combined response by males that has not beenpreviously described: males in the presence of rivals increasecopulation duration and the rate of sperm transfer. No relationshipwas found between male or female size and copulation durationor ejaculate size. Golden egg bug males transfer sperm slowlyand gradually throughout copulation; thus an increase in theamount of sperm transferred and the corresponding increase inthe male's numerical representation in the female's storageorgans could be particularly important in a system in whichso few sperm are transferred and in which so few sperm are storedby females. In addition, copulation duration may not only serveto increase the total amount of sperm transferred, but it mayalso increase the chances that the female will lay an egg soonafter copulation has ended. This could explain why males tendto accept eggs after copulation, since they could be maximizingthe chances that such eggs are fathered by them, and in thisway they would substantially increase the survival rates oftheir offspring because eggs laid on plants suffer high mortalityrates.     

Remote copulation: male adaptation to female cannibalism

Sexual cannibalism by females and associated male behaviours may be driven by sexual conflict. One such male behaviour is the eunuch phenomenon in spiders, caused by total genital emasculation, which is a seemingly maladaptive behaviour. Here, we provide the first empirical testing of an adaptive hypothesis to explain this behaviour, the remote copulation, in a highly sexually cannibalistic orb-web spider Nephilengys malabarensis. We demonstrate that sperm transfer continues from the severed male organ into female genitals after the male has been detached from copula. Remote copulation increases the total amount of sperm transferred, and thus probably enhances paternity. We conclude that the mechanism may have evolved in response to sexual cannibalism and female-controlled short copulation duration.     

One size fits all? Determinants of sperm transfer in a highly dimorphic orb‐web spider

The evolutionary significance of widespread hypo‐allometric scaling of genital traits in combination with rapid interspecific genital trait divergence has been of key interest to evolutionary biologists for many years and remains poorly understood. Here, we provide a detailed assessment of quantitative genital trait variation in males and females of the sexually highly dimorphic and cannibalistic orb‐weaving spider Argiope aurantia. We then test how this trait variation relates to sperm transfer success. In particular, we test specific predictions of the one‐size‐fits‐all and lock‐and‐key hypotheses for the evolution of genital characters. We use video‐taped staged matings in a controlled environment with subsequent morphological microdissections and sperm count analyses. We find little support for the prediction of the one‐size‐fits‐all hypothesis for the evolution of hypo‐allometric scaling of genital traits, namely that intermediate trait dimensions confer highest sperm transfer success. Likewise, our findings do not support the prediction of the lock‐and‐key hypothesis that a tight fit of male and female genital traits mediates highest sperm transfer success. We do, however, detect directional effects of a number of male and female genital characters on sperm transfer, suggesting that genital trait dimensions are commonly under selection in nature. Importantly, even though females are much larger than males, spermatheca size limits the number of sperm transferred, contradicting a previous hypothesis about the evolutionary consequences of genital size dimorphism in extremely size‐dimorphic taxa. We also find strong positive effects of male body size and copulation duration on the probability of sperm transfer and the number of sperm transferred, with implications for the evolution of extreme sexual size dimorphism and sexual cannibalism in orb weavers.     

A potential mechanism for cryptic female choice in a flour beetle

In polyandrous mating systems, events occurring during copulation can alter the fate of the mating male's sperm. These events may exert selective pressures resulting in the evolution of diverse reproductive behaviours, morphologies and physiologies. This study investigates the role of male and female copulatory behaviours on sperm fate in the red flour beetle, Tribolium castaneum. I describe and quantify copulatory behaviours for mating pairs, and examine sperm fate by quantifying sperm transfer, sperm storage and second male sperm precedence. Whereas female quiescence during copulation and male leg rubbing during copulation together account for significant variation (26%) in sperm precedence, female copulatory quiescence alone provides information about the timing and numbers of sperm transferred. These experiments show that a female copulatory behaviour predicts sperm fate, and emphasize the value of studying variation in both male and female copulatory behaviours for understanding factors affecting sperm use.     

Mating behavior,insemination and sperm transfer in the ground beetle Carabus insulicola

Mating behavior and the processes of insemination and sperm transfer in the ground beetle Carabus insulicola were analyzed. C. insulicola has elaborate genitalia, in which the strongly sclerotized male copulatory piece is inserted into the female vaginal appendix in copula. During mating, I observed pre-copulatory struggles of males and females, as well as delays in ejaculation, suggesting the presence of intersexual conflicts. Insemination was achieved with a spermatophore, which strongly adhered to the openings of the spermatheca, common oviduct, and vaginal appendix. The spermatophore dissolved after copulation, and sperm were transferred into the spermatheca within three hours after copulation. Sperm bundles were contained within the testes and spermatophores, but free spermatozoa were found in the spermatheca.