Opportunities for female choice in the bed bug Cimex lectularius

Bed bugs are cited as exemplars of sexual conflict because mating can only occur via traumatic insemination. However, past antagonistic coevolution between the sexes does not necessarily preclude current female choice. Here, we investigate opportunities for precopulatory female choice in bed bugs. We examined whether females seek out mating opportunities when they gain the most benefit: when females are virgin and/or have recently fed. But, we found that female mating and feeding status had little effect on female attraction to males and male odor. To determine whether females approach male harborages (home crevices) to seek matings in nature, we investigated where matings occurred among unfamiliar pairs of bed bugs. We found that, despite female attraction to male odor, matings were most likely to take place in the female's harborage rather than the male's harborage. We also examined the effect of feeding on male and female ability to mate. Whereas previous research reported that engorgement impaired female ability to refuse matings, we found that male feeding status had a larger effect on the success of mating encounters than female feeding status. Fed males had poor mating success, suggesting that males may be faced with a trade‐off between mating and feeding.     

Reducing a cost of traumatic insemination: female bedbugs evolve a unique organ

The frequent wounding of female bedbugs (Cimex lectularius: Cimicidae) during copulation has been shown to decrease their fitness, but how females have responded to this cost in evolutionary terms is unclear. The evolution of a unique anatomical structure found in female bedbugs, the spermalege, into which the male's intromittent organ passes during traumatic insemination, is a possible counteradaptation to harmful male traits. Several functions have been proposed for this organ, and we test two hypotheses related to its role in sexual conflict. We examine the hypotheses that the spermalege functions to (i) defend against pathogens introduced during traumatic insemination; and (ii) reduce the costs of wound healing during traumatic insemination. Our results support the 'defence against pathogens' hypothesis, suggesting that the evolution of this unique cimicid organ resulted, at least partly, from selection to reduce the costs of mating-associated infection. We found no evidence that the spermalege reduces the costs of wound healing.     

Multiple mating in the traumatically inseminating Warehouse pirate bug, Xylocoris flavipes: effects on fecundity and longevity

Optimal mating frequencies differ between sexes as a consequence of the sexual differentiation of reproductive costs per mating, where mating is normally more costly to females than males. In mating systems where sexual reproduction is costly to females, sexual conflict may cause both direct (i.e. by reducing female fecundity or causing mortality) and indirect (i.e. increased risk of mortality, reduced offspring viability) reductions in lifetime reproductive success of females, which have individual and population consequences. We investigated the direct and indirect costs of multiple mating in a traumatically inseminating (TI) predatory Warehouse pirate bug, Xylocoris flavipes (Reuter) (Hemiptera: Anthocoridae), where the male penetrates the female's abdomen during copulation. This study aimed to quantify the effects of TI on female fecundity, egg viability, the lifetime fecundity schedule, longevity and prey consumption in this cosmopolitan biocontrol agent. We found no difference in the total reproductive output between mating treatments in terms of total eggs laid or offspring viability, but there were significant differences found in daily fecundity schedules and adult longevity. In terms of lifetime reproduction, female Warehouse pirate bugs appear to be adapted to compensate for the costs of TI mating to their longevity.     

Trauma, disease and collateral damage: conflict in cimicids

The bed bugs and bat bugs (Hemiptera: Cimicidae) are unusual in being a gonochorist (separate male and female genders) taxon with obligate traumatic insemination. Males of all the species in this family have a lanceolate paramere (intromittent organ) which they use to pierce the female's body wall and inseminate directly into her haemocoel, despite the presence of a functional female genital tract. Mating is tightly linked to the feeding cycle in Cimex lectularius, the common bed bug. In this paper, I examine key aspects of the reproductive anatomy and behaviour of C. lectularius that underpin the nature of the conflict over mating rate in this species. I then examine the consequences of traumatic insemination for female fitness and examine potential mechanisms that might underpin those costs. Finally, the collateral consequences of the male reproductive tactic on other males of C. lectularius and the African bat bug, Afrocimex constrictus are examined.     

Sexually antagonistic coevolution in insects is associated with only limited morphological diversity

Morphological traits involved in male-female sexual interactions, such as male genitalia, often show rapid divergent evolution. This widespread evolutionary pattern could result from sustained sexually antagonistic coevolution, or from other types of selection such as female choice or selection for species isolation. I reviewed the extensive but under-utilized taxonomic literature on a selected subset of insects, in which male-female conflict has apparently resulted in antagonistic coevolution in males and females. I checked the sexual morphology of groups comprising 500-1000 species in six orders for three evolutionary trends predicted by the sexually antagonistic coevolution hypothesis: males with species-specific differences and elaborate morphology in structures that grasp or perforate females in sexual contexts; corresponding female structures with apparently coevolved species-specific morphology; and potentially defensive designs of female morphology. The expectation was that the predictions were especially likely to be fulfilled in these groups. A largely qualitative overview revealed several surprising patterns: sexually antagonistic coevolution is associated with frequent, relatively weak species-specific differences in males, but male designs are usually relatively simple and conservative (in contrast to the diverse and elaborate designs common in male structures specialized to contact and hold females in other species, and also in weapons such as horns and pincers used in intra-specific battles); coevolutionary divergence of females is not common; and defensive female divergence is very uncommon. No cases were found of female defensive devices that can be facultatively deployed. Coevolutionary morphological races may have occurred between males and females of some bugs with traumatic insemination, but apparently as a result of female attempts to control fertilization, rather than to reduce the physical damage and infections resulting from insertion of the male's hypodermic genitalia. In sum, the sexually antagonistic coevolution that probably occurs in these groups has generally not resulted in rapid, sustained evolutionary divergence in male and female external sexual morphology. Several limitations of this study, and directions for further analyses are discussed.     

Potential sexual transmission of environmental microbes in a traumatically inseminating insect

Abstract.  1. Very little attention has been paid to the importance of environmental microbes and how they gain access to potential hosts during host wounding or sexual interactions. Male members of the family Cimicidae (including the haematophagous human pest, the bed bug, Cimex lectularius ) show traumatic insemination: in order to transfer sperm the male punctures the female's cuticle using a needle-like intromittent organ. The microbes from the local environment that gain entry into females during mating have previously been shown to be an important source of mortality. This study aimed to identify these microbes and their natural environmental substrates.
2. Nine different microbe taxa from the surface of the cuticle were identified, five of them (two Penicillium spp., Stenotrophomonas , Enterobacter , Bacillus ) were found in the local environment as well as on the intromittent organ of male bed bugs: these are strong candidates for the causal agents of mating-induced female mortality. Local differences in the microbial communities and their hosts' response to them are discussed as a source of reproductive isolation.
3. Four microbe genera ( Scopulariopsis , Staphylococcus , Arthrobacter , Micrococcus ) were found exclusively on blood agar. One of them, and four grown on regular agar, are classified as human pathogens. However, because no microbes were isolated from the piercing and sucking mouthparts the epidemiological significance of bed bugs carrying externally attached microbes is probably low.     

The Effect of Abdominal Spines on Female Mating Frequency and Fecundity in a Water Strider

Conflict between the sexes over mating decision may result in antagonistic coevolution in structures that increase control over copulation. In Aquarius paludum both females and males have long abdominal spines. We tested the hypothesis that abdominal spines increase female ability to resist male mating attempts and reduce the costs of mating in A. paludum. We manipulated female spine length and observed female mating and egg-production rate in two different studies. We found that females with intact spines succeeded to reject male mating attempt more often than females with removed spines. Intact females also mated less often than females with removed or shortened spines. Male presence and mating rate increased female egg number. Our results thus support the hypothesis that abdominal spines help female to reject male mating attempts but contrary to predictions, we found that A. paludum females somehow benefit from multiple matings in spite of the sexual conflict.     

The evolution of polyandry: multiple mating and female fitness in insects

Theory suggests that male fitness generally increases steadily with mating rate, while one or a few matings are sufficient for females to maximize their reproductive success. Contrary to these predictions, however, females of the majority of insects mate multiply. We performed a meta-analysis of 122 experimental studies addressing the direct effects of multiple mating on female fitness in insects. Our results clearly show that females gain directly from multiple matings in terms of increased lifetime offspring production. Despite a negative effect of remating on female longevity in species without nuptial feeding, the positive effects (increased egg production rate and fertility) more than outweigh this negative effect for moderate mating rates. The average direct net fitness gain of multiple mating was as high as 30-70%. Therefore, the evolutionary maintenance of polyandry in insects can be understood solely in terms of direct effects. However, our results also strongly support the existence of an intermediate optimal female mating rate, beyond which a further elevated mating rate is deleterious. The existence of such optima implies that sexual conflict over the mating rate should be very common in insects, and that sexually antagonistic coevolution plays a key role in the evolution of mating systems and of many reproductive traits. We discuss the origin and maintenance of nuptial feeing in the light of our findings, and suggest that elaborate and nutritional ejaculates may be the result of sexually antagonistic coevolution. Future research should aim at gaining a quantitative understanding of the evolution of female mating rates. Copyright 2000 The Association for the Study of Animal Behaviour.     

Nymphs of the common bed bug (<Emphasis Type="Italic">Cimex lectularius</Emphasis>) produce anti-aphrodisiac defence against conspecific males

Background  

Abdominal wounding by traumatic insemination and the lack of a long distance attraction pheromone set the scene for unusual sexual signalling systems. Male bed bugs (Cimex lectularius) mount any large, newly fed individual in an attempt to mate. Last instar nymphs overlap in size with mature females, which make them a potential target for interested males. However, nymphs lack the female's specific mating adaptations and may be severely injured by the abdominal wounding. We, therefore, hypothesized that nymphs emit chemical deterrents that act as an honest status signal, which prevents nymph sexual harassment and indirectly reduces energy costs for males.     

Female sexual polymorphism and fecundity consequences of male mating harassment in the wild

Genetic and phenotypic variation in female response towards male mating attempts has been found in several laboratory studies, demonstrating sexually antagonistic co-evolution driven by mating costs on female fitness. Theoretical models suggest that the type and degree of genetic variation in female resistance could affect the evolutionary outcome of sexually antagonistic mating interactions, resulting in either rapid development of reproductive isolation and speciation or genetic clustering and female sexual polymorphisms. However, evidence for genetic variation of this kind in natural populations of non-model organisms is very limited. Likewise, we lack knowledge on female fecundity-consequences of matings and the degree of male mating harassment in natural settings. Here we present such data from natural populations of a colour polymorphic damselfly. Using a novel experimental technique of colour dusting males in the field, we show that heritable female colour morphs differ in their propensity to accept male mating attempts. These morphs also differ in their degree of resistance towards male mating attempts, the number of realized matings and in their fecundity-tolerance to matings and mating attempts. These results show that there may be genetic variation in both resistance and tolerance to male mating attempts (fitness consequences of matings) in natural populations, similar to the situation in plant-pathogen resistance systems. Male mating harassment could promote the maintenance of a sexual mating polymorphism in females, one of few empirical examples of sympatric genetic clusters maintained by sexual conflict.     

Costly sexual harassment in a beetle

Abstract The optimal number of mating partners for females rarely coincides with that for males, leading to sexual conflict over mating frequency. In the bruchid beetle Callosobruchus maculatus, the fitness consequences to females of engaging in multiple copulations are complex, with studies demonstrating both costs and benefits to multiple mating. However, females kept continuously with males have a lower lifetime egg production compared with females mated only once and then isolated from males. This reduction in fitness may be a result of damage caused by male genitalia, which bear spines that puncture the female’s reproductive tract, and/or toxic elements in the ejaculate. However, male harassment rather than costs of matings themselves could also explain the results. In the present study, the fitness costs of male harassment for female C. maculatus are estimated. The natural refractory period of females immediately after their first mating is used to separate the cost of harassment from the cost of mating. Male harassment results in females laying fewer eggs and this results in a tendency to produce fewer offspring. The results are discussed in the context of mate choice and sexual selection.     

Comparative transcriptomics of the bacteriome and the spermalege of the bedbug Cimex lectularius (Hemiptera: Cimicidae)

The bedbug Cimex lectularius is notorious as a blood-feeding exoparasite of human and other warm-blooded animals. In addition to its medical and hygienic importance, C. lectularius is known for its unique biological traits, including obligate nutritional mutualism with a vitamin-provisioning Wolbachia endosymbiont and a peculiar mating habit called traumatic insemination wherein male sperm is injected into the female body cavity, migrates to the ovary, and fertilizes eggs therein. For these unique traits, novel insect organs, the bacteriome for endosymbiosis and the spermalege for traumatic insemination, have evolved in the lineage of bedbugs. We constructed cDNA libraries of the bacteriome and the spermalege of C. lectularius, and performed expressed sequence tag (EST) analyses of these peculiar insect organs. In total 4480 ESTs were compiled, which were assembled into 2989 unique sequences (USs). The following ESTs and USs were identified from the different organs: 1151 ESTs and 901 USs from the female whole body; 1098 ESTs and 879 USs from the female bacteriome; 1145 ESTs and 783 USs from the male bacteriome; and 1086 ESTs and 920 USs from the female spermalege. These EST data will provide a valuable genetic resource for understanding the developmental and evolutionary aspects of these novel insect organs.     

Is egg carrying attractive? Mate choice in the golden egg bug (Coreidae,Heteroptera)

In several species of fish, females select males that are already guarding eggs in their nests. It is a matter of debate as to whether a female selects a good nest site for her offspring (natural selection) or a male for his attractiveness (sexual selection). The golden egg bug, Phyllomorpha laciniata Vill, resembles fish in the sense that mating males carry more eggs than single males, but in the bugs, female mate choice is decoupled from egg site choice. The sexual selection hypothesis predicts that if females select males using male egg load as a cue for male quality, they should not mate with a male when eggs are removed, regardless of his mating attempts. When individual females were enclosed with an egg-loaded male and an unloaded male, they mated equally often with both males, although the loaded males courted more. In addition, when only successful males were used, females mated equally often with the loaded male and the unloaded male irrespective of sex ratio. Male choice rather than female choice affected mating frequency when sex ratio was equal. Therefore, females do not select the male by the eggs he carries, but successful males may receive many eggs due to egg dumping by alien females while they mate or as a consequence of mate guarding.     

Harm to females increases with male body size in Drosophila melanogaster

Previous studies indicate that female Drosophila melanogaster are harmed by their mates through copulation. Here, we demonstrate that the harm that males inflict upon females increases with male size. Specifically, both the lifespan and egg-production rate of females decreased significantly as an increasing function of the body size of their mates. Consequently, females mating with larger males had lower lifetime fitness. The detrimental effect of male size on female longevity was not mediated by male effects on female fecundity, egg-production rate or female-remating behaviour. Similarly, the influence of male size on female lifetime fecundity was independent of the male-size effect on female longevity. There was no relationship between female size and female resistance to male harm. Thus, although increasing male body size is known to enhance male mating success, it has a detrimental effect on the direct fitness of their mates. Our results indicate that this harm is a pleiotropic effect of some other selected function and not an adaptation. To the extent that females prefer to mate with larger males, this choice is harmful, a pattern that is consistent with the theory of sexually antagonistic coevolution.     

A phylogenetic analysis of the evolution of sexual dimorphism and mating systems in water striders (Hemiptera: Gerridae)

Conflicts over mating decisions characterize the sexual behaviour of many insects, in particular when males encounter females that already carry enough sperm to fertilize their eggs, since a mating often will inflict greater costs than benefits upon females. Therefore, coevolutionary models predict adaptation and counter-adaptation by the sexes in a battle to control the outcome of sexual encounters. A phylogenetic analysis was performed on patterns of sexual dimorphism and mating systems within water striders (Hemiptera, Gerridae). Phylogenetic effects or 'constraints' have significantly shaped patterns of sexual dimorphism in female/ male size ratios, legs and genitalia of males, and the structure of the female abdomen. Males of ancestral gerrids were probably slightly smaller than conspecific females, had powerful fore legs adapted to grasp the female's thorax during mating, and had clasping genitalic structures suited to grasp or pinch the female posteriorly. Most gerrids have a female/male size ratio between 1.05 and 1.14, but more pronounced sexual size ratios (above 1.25) have independently evolved several times in the family, usually in association with extended post-copulatory mate guarding. The comparative, phylogenetic analysis suggests coevolution of female anticlasper and male clasping devices for the clade comprising the subfamilies Cylindrostethinae, Ptilomerinae, and Halobatinae while female anticlasper devices have evolved in the absence of male clasping genitalia in the Gerrinae. The ancestral and most common mating system in gerrids is 'scramble competition polygyny' from which has evolved 'resource defence polygyny' at least four times independently of each other. The phylogenetic effects on patterns of mating behaviour are much less obvious, as exemplified by the large amount of interspecific variation in some genera.     

Sexual conflict and antagonistic coevolution across water strider populations

Microevolutionary studies have demonstrated sexually antagonistic selection on sexual traits, and existing evidence supports a macroevolutionary pattern of sexually antagonistic coevolution. Two current questions are how antagonistic selection within-populations scales to divergence among populations, and to what extent intraspecific divergence matches species-level patterns. To address these questions, we conducted an intraspecific comparative study of sexual armaments and mating behaviors in a water strider (Gerris incognitus) in which male genitals grasp resistant females and female abdominal structures help ward off males. The degree of exaggeration of these armaments coevolves across species. We found a similar strong pattern of antagonistic coevolution among populations, suggesting that sexual conflict drives population differentiation in morphology. Furthermore, relative exaggeration in armaments was closely related to mating outcomes in a common environment. Interestingly, the effect of armaments on mating was mediated by population sexual size dimorphism. When females had a large size advantage, mating activity was low and independent of armaments, but when males had a relative size advantage, mating activity depended on which sex had relatively exaggerated armaments. Thus, a strong signal of sexually antagonistic coevolution is apparent even among populations. These results open opportunities to understand links between sexual arms races, ecological variation, and reproductive isolation.     

Sexual conflict and cooperation in butterfly reproduction: a comparative study of polyandry and female fitness

Most butterfly species can be characterized as capital breeders, meaning that reproductive output is strongly coupled to the amount of resources they have procured during the larval stage. Accordingly, female fecundity is generally correlated with female mass, both within and across species. However, the females of some species can be partly characterized as income breeders, in the sense that their reproductive output is dependent not only on larval-derived capital but also on resources acquired during the adult stage. These adult resources can be derived from female feeding or from male-transferred nuptial gifts. Recent studies on the within-species effects of multiple matings on female fitness show that females generally gain directly from multiple matings in terms of increased lifetime offspring production. Here, we test whether the positive effects of multiple mating on female fitness also hold at a comparative level, by conducting a laboratory study of female reproductive output in eight pierid species that differ in life-time female mating frequency. Female reproductive output, measured as cumulative egg mass divided by female mass, increased significantly with polyandry (r = 0.942, p < 0.001), demonstrating that the positive effect of mating rate on female reproductive fitness also holds between species. The positive effect of male nutrient contribution is substantial, and the per capita reproductive output is more than twice as high in the most polyandrous species as in the most monandrous ones. Hence, the positive net effect of the ejaculates is highly substantial, although males and females can have sexual interests that run counter to each other, setting the stage for sexually antagonistic coevolution, so that the various component parts of the male ejaculate-sperm, nutrients, anti-aphrodisiacs, and gonadotrophic hormones-may each correspond to a separate conflict-cooperation balance between the sexes. Two scenarios for the evolution of nuptial gifts in butterflies are discussed, one arguing that variation in larval food is the underlying factor and the other arguing that sexually antagonistic coevolution is the driving force. The two views are complementary rather than mutually exclusive, although the former hypothesis predicts that variation in female mating rate depends on variation in larval food availability, whereas the latter suggests that variation in female mating rate between species results from species-specific idiosyncrasies.     

Mating behaviour and mating success in woodfrogs Rana sylvatica

Mating behaviour of woodfrogs was investigated in a woodland pond in southeastern Michigan. Males became sexually mature one year before females and outnumbered females by 5.6 to 1 at the breeding site. Males employed searching behaviour to obtain mates. Yearly male mating success varied from 0 to 2 matings, and larger males had a greater probability of mating than smaller males. The mating season lasted less than 10 days. Most egg masses were deposited in 1 m² of a 256-m² pond. Experimental introduction of egg masses to the breeding site indicated that presence of eggs was sufficient to stimulate egg deposition. The influence of selection for synchronous sexual receptivity in females, communal egg deposition, and delayed female sexual maturation on male mating behaviour and variation in male mating success is discussed.     

Sisters' curse: sexually antagonistic effects constrain the spread of a mitochondrial haplogroup superior in sperm competition

Maternal inheritance of mitochondria creates a sex-specific selective sieve with implications for male longevity, disease susceptibility and infertility. Because males are an evolutionary dead end for mitochondria, mitochondrial mutations that are harmful or beneficial to males but not females cannot respond directly to selection. Although the importance of this male/female asymmetry in evolutionary response depends on the extent to which mitochondrial mutations exert antagonistic effects on male and female fitness, few studies have documented sex-specific selection acting on mitochondria. Here, we exploited the discovery of two highly divergent mitochondrial haplogroups (A and B2) in central Panamanian populations of the pseudoscorpion Cordylochernes scorpioides. Next-generation sequencing and phylogenetic analyses suggest that selection on the ND4 and ND4L mitochondrial genes may partially explain sexually antagonistic mitochondrial effects on reproduction. Males carrying the rare B2 mitochondrial haplogroup enjoy a marked advantage in sperm competition, but B2 females are significantly less sexually receptive at second mating than A females. This reduced propensity for polyandry is likely to significantly reduce female lifetime reproductive success, thereby limiting the spread of the male beneficial B2 haplogroup. Our findings suggest that maternal inheritance of mitochondria and sexually antagonistic selection can constrain male adaptation and sexual selection in nature.     

Mating and social contact change egg production and longevity in adult females of the mirid Lygus hesperus

There is frequently a tradeoff between fecundity and longevity, but the relationship is inconsistent across species and influenced by various exogenous and endogenous factors. Previous studies of Lygus hesperus Knight (Hemiptera: Miridae) established that egg production is promoted by insemination, at least temporarily, but little is known about the long‐term effects of mating and nonsexual interactions with conspecifics on egg production and female lifespan. To elucidate these relationships, survivorship and oviposition rate were tracked daily in females that were isolated or paired with a fertile male or another female throughout their adult lives. Mating rates were determined by postmortem examination. Results indicate that male‐specific stimuli accelerate female reproductive maturation, and that mating elevates oviposition rate. However, females paired with either a female or male companion had shortened lifespans, suggesting that social contacts exact a significant cost in this solitary species. Despite the negative impact of conspecific interactions and the finding that a singly mated female has sufficient sperm to fertilize a lifetime supply of eggs, many females were found to have mated more than once. Multiply mated females had higher sustained oviposition rates, lived longer, and had greater lifetime fecundities. Collectively, no strong evidence was found of a direct physiological link between fecundity and longevity, but environmental factors and mating were found to significantly influence both traits.