7‐methylheptacosane is a major component of the contact sex pheromone of the cerambycid beetle Neoclytus acuminatus acuminatus

Abstract Male Neoclytus acuminatus acuminatus (F.) (Coleoptera: Cerambycidae) attempt to mate with females only after touching them with their antennae, suggesting that mate recognition is mediated by contact pheromones in the cuticular wax layer of females. Consistent with that hypothesis, males exhibit similar responses to dead females in laboratory bioassays, but not to solvent‐washed dead females with their cuticular hydrocarbons removed. The mating response of males is restored when solvent extracts are reapplied to carcasses of solvent‐washed females, indicating that the contact pheromone is present in solvent extracts. Solvent extracts of the female cuticle contain six methylalkanes that are not present in extracts of males, three of which (7Me‐C₂₅, 7Me‐C₂₇ and 9Me‐C₂₇) constitute almost 40% of the total hydrocarbons. The bioactivity of these three compounds is tested by applying synthetic standards to solvent‐washed carcasses of females and presenting them to males. Standards are tested singly, pairwise and as the complete blend; freeze‐killed females serve as controls. Males attempt to couple with solvent‐washed female carcasses treated with 7Me‐C₂₇ alone and in combination with 9Me‐C₂₇ but only the complete blend elicits the same number of mounting and coupling attempts as does the control. These findings suggest that 7Me‐C₂₇ (7‐methylheptacosane) is the major component of the contact sex pheromone of N. a. acuminatus and that 7Me‐C₂₅ and 9Me‐C₂₇ act as synergists.     

Mating behavior and contact pheromones of Batocera horsfieldi (Hope) (Coleoptera: Cerambycidae)

The mating behavior of Batocera horsfieldi (Hope) was observed in the laboratory. The results showed that copulation of this longhorned beetle consisted of three phases: (i) encountering and pair‐bonding; (ii) mating attempt and ejaculation; and (iii) post‐copulatory guarding. Frozen females, with cuticular hydrocarbons stripped by hexane extraction, showed no attraction for males. Reapplying the solvent extract of frozen females to both washed dead males and females caused mating attempts by males, confirming that cuticular hydrocarbons (contact sex pheromones) played an important role in recognition of females by male B. horsfieldi.     

A female‐produced short‐range sex pheromone in the egg parasitoid Trissolcus brochymenae

The potential for short‐range sex pheromone communication by the egg parasitoid wasp Trissolcus brochymenae (Hymenoptera: Platygastridae) was investigated in closed arena bioassays. Males of this parasitoid showed more antennal drumming and more frequent mounting behaviour on 1‐ to 2‐d‐old virgin females compared with 8‐d‐old virgin females. Male copulation attempts were fewer with previously mated females than with virgin females. Males courted and made copulation attempts with 1‐ to 2‐d‐old female cadavers, but not with male cadavers or with female cadavers rinsed in organic solvents of different polarities. Male attraction to female cadavers was re‐established by treating cadavers with acetone extracts of females, but not with ether or hexane extracts. In experiments using female cadavers dissected into head, mesosoma, and gaster, and then reassembled using one unwashed body section and two body sections washed in acetone, males were attracted only to the reassembled cadavers with an unwashed mesosoma. These findings suggest that (1) courtship behaviour in males of T. brochymenae is triggered by a short‐range sex pheromone produced by females; (2) the age and the physiological condition of females (virgin/mated) influence pheromone release or production; (3) the female's mesosoma is the source of the sex pheromone; and (4) polar components of the sex pheromone play a major role in influencing male behaviour. Our results suggest that quasi‐gregarious egg parasitoids are selected for short‐range rather than long‐range sex pheromones.     

Male chemical cues as reliable indicators of infection in the wolf spider Schizocosa ocreata

Sexual signals can convey important information about mate quality, such as critical information about a signaler's health status, helping an individual to avoid infected or immunocompromised conspecifics. Chemical signals are especially important in this context, because they represent an honest and dynamic signaling modality that receivers can use to make updated mate choice decisions to avoid compromising their own health. In this study, we investigated the viability of male chemical cues in the wolf spider Schizocosa ocreata as a reliable indicator of health status. Using video playback with images of an average male that is simultaneously paired with male cuticular compounds on filter paper, we show that females are more receptive to videos paired with cues from control males rather than infected males. We also show that these cuticular compounds can be isolated and retain similar female behavioral responses when extracted with a nonpolar solvent, suggesting that these cuticular compounds may not be just complex hydrocarbons, but a combination of cuticular compounds. This is the first evidence for female discrimination and recognition of male chemical cues in this species, which opens up important new avenues of research in a well‐studied species with complex multimodal signaling.     

Discrimination of conspecific individuals via cuticular pheromones by males of the cricket Gryllus bimaculatus

Cuticular substances on the body surface of crickets serve as pheromones that elicit a variety of different behaviors in male crickets. Antennal contact between males and females resulted in courtship behavior, and that between two males resulted in aggressive displays. As a first step in elucidating how crickets recognize and discriminate individuals, behavioral responses of male individuals to cuticular substances of conspecific males or females were investigated. The behavioral responses of males to antennal or palpal stimulation with an isolated antenna from a male or a female were recorded. To both antennal and palpal stimulation with female antennae, the majority of males responded with courtship behavior; to stimulation with male antennae, males responded with aggressive displays. To gain insight into the chemical nature of the behaviorally relevant components, isolated antennae were washed in either n-hexane, acetone or ethanol before behavior assays. Washed antennae no longer elicited courtship or aggressive responses in males. Next, polypropylene fibers were smeared with substances from the body surface of females and used for antennal stimulation. This experiment showed that the quality and quantity of cuticular substances appear to be highly age-dependent. Significantly more males responded with courtship behavior to cuticular substances from younger females. Isolated males generally showed higher levels of aggression than males reared in groups. Grouped males also were more likely to display courtship behavior towards antennae from younger females, and aggressive behavior towards antennae from older females. These results suggest that male discrimination of mating partners depends on the nature of female cuticular substances.     

Contact chemoreceptive mate recognition in Cerambyx welensii Küster (Coleoptera: Cerambycidae)

1. Cerambyx welensii Küster (Coleoptera: Cerambycidae) is a wood-borer responsible for the decline of Mediterranean oaks in open woodlands.
2. To establish that contact pheromones are involved in mate recognition of C. welensii, we extracted the cuticular hydrocarbons by solid phase microextraction at pre-reproductive, beginning and ending of the reproductive period, and by solvent extraction of prothorax and elytra. The extracts were analysed by GC–MS under electron impact and chemical ionization conditions. Cuticular hydrocarbon profiles varied according to the reproductive period, although differences between sexes were not significant. Two compounds, 11/13-methylheptacosane and 11-methylnonacosane were more abundant in females at the beginning and ending of the reproductive periods. Compound 11/13-methylheptacosane was also more representative in female prothorax than in males, and 2-methyloctacosane was richer in male elytra than in females.
3. We also studied the role of cuticular hydrocarbons in mate recognition in arena bioassays. Treatment of solvent-washed dead females and glass dummies with one female equivalent (FE) of cuticular extract elicited mating responses in males, especially at the beginning of the reproductive period, with copulation attempts reaching 61.9% on solvent-washed dead females and 23.8% on dummies. The successive treatment with synthetic compounds approaching a male cuticular profile inhibited male response.
4. Our results confirm that contact pheromones mediate mate recognition in C. welensii. Knowledge of the precise role played by the major compounds 11/13-methylheptacosane and 11-methylnonacosane and other minor compounds representative in female prothorax may contribute to the development of novel management strategies against C. welensii.

     

The Coolidge effect, individual recognition and selection for distinctive cuticular signatures in a burying beetle

The ability to recognize individuals is an important aspect of social interactions, but it can also be useful to avoid repeated matings with the same individual. The Coolidge effect is the progressive decline in a male's propensity to mate with the same female combined with a heightened sexual interest in new females. Although males that recognize previous partners and show a preference for novel females should have a selective advantage as they can distribute sperm evenly among the females they encounter, there are few invertebrate examples of the Coolidge effect. Here we present evidence for this effect in the burying beetle Nicrophorus vespilloides and examine the mechanism underlying the discrimination between familiar and novel mates. Burying beetles feed and reproduce on vertebrate carcasses, where they regularly encounter conspecifics. Males showed greater sexual interest in novel females (virgin or mated) than in females they had inseminated before. The application of identical cuticular extracts allowed us to experimentally create females with similar odours, and male responses to such females demonstrated that they use female cuticular patterns for discrimination. The chemical analysis of the cuticular profile revealed greater inter-individual variation in female than in male cuticular patterns, which might be due to greater selection on females to signal their individual identity.     

Morphometric variation in bluegill Lepomis macrochirus and green sunfish Lepomis cyanellus in lentic and lotic systems

Bluegill Lepomis macrochirus and green sunfish Lepomis cyanellus were examined using geometric morphometrics to evaluate the variation in morphology between fishes that reside in lentic (e.g. lakes) and lotic (e.g. streams) ecosystems. Live fishes were collected from reservoirs and rivers in central Indiana, while additional fishes were sampled from museum collections at Ball State University and the Illinois Natural History Survey. Male and female L. macrochirus and female L. cyanellus from lentic systems display a deeper body than those from lotic systems, while no differences were found in male L. cyanellus morphometry. A deeper body promotes greater manoeuverability, typically desirable in lentic systems. In contrast, the more streamlined body of the fishes found in lotic systems reduces drag as it contends with flowing water, ultimately maximizing energy efficiency. The absence of morphological differences, such as those found in male L. cyanellus, may be caused by fish occupying both lentic and lotic systems, from the population not having been present in the body of water long enough to display any adaptations, or from a lack of statistical power caused by the small sample size.     

Observations on the mating behavior of the alfalfa blotch leafminer,Agromyza frontella (Rondani) (Diptera: Agromyzidae), and evidence of a female sex pheromone

The mating behavior of Agromyza frontella was studied under laboratory conditions. Adults were able to mate on the day of emergence, with no evident periodicity throughout the photophase. The host plant was essential for mating to occur, its presence affecting female receptivity rather than male copulatory behavior. Males generally entered a stationary phase once in the proximity of a female, before undertaking a final approach. This stationary behavior frequently resulted in male aggregations around a female, and under such conditions males exhibited a characteristic wing vibrating behavior. As male wing vibration was not an essential behavior for successful mating, and rarely occurred during male encounters in the absence of females or when only one male was near a female, it was considered as being primarily a male-male signal. The majority of females that mated exhibited an ovipositor pumping behavior that stimulated the male approach. However, such behavior was not essential to attract mates, as dead females elicited the entire sequence of male mating behavior. This suggested the presence of a cuticular sex pheromone, as reported for other species of higher Diptera. Whole virgin female (<24- h or 3- day-old) hexane extracts applied to male cadavers increased the time males spent on the plant, the number of contacts with the treated cadavers, the incidence of attempted copulations, and the wing vibrating behavior between males compared with controls using untreated cadavers. The results obtained indicate that females control copulation in A. frontella and that both semiochemical and visual cues are important in eliciting male mating behavior.     

Contest dynamics for food and reproductive resources are defined by health condition in a dung beetle

Male dung beetles compete to obtain food and females, and early resource recognition and appropriation increase the probability of mating. The outcome of such encounters is expected to be defined by self‐ and the opponent’s health condition. In this study, we analyzed the effect of body condition and immune defense on the contest dynamics between males that rolled a food ball with a partner female (owner males) and intruder males of dung beetle Canthon cyanellus. Body condition was measured as body size, body dry mass, lipid mass, and muscle mass; immune defense was estimated via phenoloxidase activity. Owner males with higher lipid mass contacted the food ball significantly earlier than owner males with lower lipid mass. Individuals with lower phenoloxidase activity started to roll food balls earlier than individuals with higher phenoloxidase activity. Owner males that had higher body dry mass, compared to female partners, began to roll the food ball significantly earlier than male–female pairs with lower differences in dry mass. Heavier males won significantly more contests than lighter males. Our results suggest that the health condition is a key factor related to the dynamics and outcome of male–male contests for resources and females in C. cyanellus. Consequently, differences in individual condition are main determinants of contest outcomes in dung beetles.     

Implications of stunting on morphology of freshwater fishes

The purpose of this study was to assess morphological differences between stunted and non‐stunted white perch Morone americana and green sunfish Lepomis cyanellus. Few female M. americana were captured; thus, morphological differences between adult males and juveniles were assessed for M. americana. Similarly, few immature (juvenile) L. cyanellus were captured for the stunted morphotype; thus, male and female morphological differences were assessed for L. cyanellus. Features of the head tended to be relatively larger in stunted fish of both species, whereas the mid‐body tended to be relatively larger in non‐stunted M. americana, but not in non‐stunted L. cyanellus. Adult and juvenile morphology overlapped considerably in non‐stunted M. americana, but there was a clear distinction between adult and juvenile morphology of stunted M. americana. There was little sexual dimorphism in shape in stunted L. cyanellus, whereas sexual dimorphism was evident in non‐stunted L. cyanellus. It appears that selective forces imposed by predation and food limitation may contribute to morphological diversification between stunted and non‐stunted fishes.     

Reproductive costs of mating with a sibling male: sperm depletion and multiple mating in Cephalonomia hyalinipennis

Polygynous parasitoid males may be limited by the amount of sperm they can transmit to females, which in turn may become sperm limited. In this study, I tested the effect of male mating history on copula duration, female fecundity, and offspring sex ratio, and the likelihood that females will have multiple mates, in the gregarious parasitoid Cephalonomia hyalinipennis Ashmead (Hymenoptera: Bethylidae: Epyrinae), a likely candidate for sperm depletion due to its local mate competition system. Males were eager to mate with the seven females presented in rapid succession. Copula duration did not differ with male mating history, but latency before a first mating was significantly longer than before consecutive matings. Male mating history had no bearing on female fecundity (number of offspring), but significantly influenced offspring sex ratio. The last female to mate with a given male produced significantly more male offspring than the first one, and eventually became sperm depleted. In contrast, the offspring sex ratio of first‐mated females was female biased, denoting a high degree of sex allocation control. Once‐mated females, whether sperm‐depleted or not, accepted a second mating after a period of oviposition. Sperm‐depleted females resumed production of fertilized eggs after a second mating. Young, recently mated females also accepted a second mating, but extended in‐copula courtship was observed. Carrying out multiple matings in this species thus seems to reduce the cost of being constrained to produce only haploid males after accepting copulation with a sperm‐depleted male. I discuss the reproductive fitness costs that females experience when mating solely with their sibling males and the reproductive fitness gain of males that persist in mating, even when almost sperm‐depleted. Behavioural observations support the hypothesis that females monitor their sperm stock. It is concluded that C. hyalinipennis is a species with a partial local mating system.     

Young Bighorn (Ovis canadensis) Males: can they Successfully Woo Females?

Mating by young males or low male‐to‐female ratios can decrease pregnancy rates and postpone birthdates in ungulates, thereby hindering population growth. Young (2.5–3.5 yr old) male bighorn (Ovis canadensis) behave differently than older males, and age, horn size, mating behavior, and social rank help determine reproductive success. We estimated birthdates in two populations of bighorn sheep in Utah, USA, to determine if mating by young males or low male‐to‐female ratios resulted in fewer young born per female, a shift in mean timing of births, or asynchronous births. When reintroduced, the Rock Canyon population consisted of four males (two each of 2.5 yr old and 1.5 yr old) and a 1 to 7.5 ratio of males (>2 yr old) to adult females (≥3.5 yr old); the Mount Nebo population consisted of four males ≤1.5 yr old and a 0 to 12 ratio of males to adult females. For both populations, the number of young born per female did not differ between the first parturition period after reintroduction (where females were impregnated by males from their source populations) and the second period of parturition (where females were impregnated by young, reintroduced males). Mean birthdates and synchrony (SD) of births did not differ for Rock Canyon (May 12, 2001 ± 4.5 d, May 14, 2002 ± 3.2 d) or Mount Nebo (May 23, 2005 ± 8.1 d, May 22, 2006 ± 10.2 d) between the first and second years following reintroduction. Mating by young males or low male‐to‐female ratios had no demonstrable effect on the number of young born per female or timing and synchrony of births in these populations.     

Pheromones exchanged during mating: a mechanism for mate assessment in Drosophila

Many insect species are sexually dimorphic for volatile compounds associated with the cuticle. In these species, females may acquire volatile compounds from males through direct contact with males during mating. In some cases, these compounds function as antiaphrodisiacs, which inhibit subsequent male precopulatory behaviour. During mating, males may also acquire female compounds, which normally stimulate males to court. Contact during mating thus results in a mutual exchange of compounds usually found primarily, or exclusively, on one or the other sex. Drosophila melanogaster is a sexually dimorphic dipteran species in which predominant male and female cuticular hydrocarbons are mutually exchanged during mating. Using synthetic compounds, the effect of this exchange on the post-mating sexual attractiveness of both males and females was tested. The sex-predominant hydrocarbons acquired during mating decreased the attractiveness of both males and females to members of the other sex. Mated females are courted less actively than virgins, and are usually unwilling to re-mate. Similarly, recently mated males are less attractive to females than virgin males are, and may be less fertile. Thus, cuticular hydrocarbons acquired by the other sex during mating could allow both males and females to assess the immediate reproductive potential of prospective mates.     

Trade‐offs in female signal apparency to males offer alternative anti‐harassment strategies for colour polymorphic females

Colour polymorphisms are known to influence receiver behaviour, but how they affect a receiver's ability to detect and recognize individuals in nature is usually unknown. I hypothesized that polymorphic female damselflies represent an evolutionary stable strategy, maintained by trade‐offs between the relative apparency of morphs to male receivers. Using field experiments on Enallagma hageni and focal studies of E. hageni and Enallagma boreale, I tested for the first time the predictions that (i) green heteromorphs and blue andromorphs gain differential protection from sexual harassment via background crypsis and sexual mimicry, respectively, and (ii) female morphs behaviourally optimize their signal apparency to mate‐searching males. First, based on male reactions elicited by females, against a high‐contrast background, the two morphs did not differ in being detected by males, and once detected, they did not differ in being recognized (eliciting sexual reactions). However, on green ferns, heteromorphs were less likely to be detected (elicited only fly‐bys) than andromorphs, but once detected, the morphs did not differ in being recognized. In contrast, when perched on a dowel with two male signal distractors, andromorphs were detected less often, and once detected, they were recognized less often than heteromorphs. Second, in fields where females foraged, andromorphs perched higher on vegetation than heteromorphs and were more often in the vicinity of males. Neither harassment rates nor evasive behaviours differed between morphs. Males aggregated in high density near shore where solitary females were rare. Equilibrium frequencies of these and other colour morphs should reflect the relative ease with which receivers detect and recognize them in the context where they are encountered.     

Female Choice by Scent Recognition in the Spotted Cucumber Beetle

In species that demonstrate female choice, geographically distinct populations can vary in their signal‐response behaviors as a result of environmental differences or genetic drift. Observing whether or not females discriminate against males from allopatric populations can establish such signal deviations. Here we compare mating success within and between populations of the spotted cucumber beetle (Diabrotica undecimpunctata howardi) collected from Delaware, Tennessee, Missouri, and New Mexico, USA. A no‐choice cross‐mating experiment was employed to measure female preference for sympatric and allopatric males. While only two of the populations (Tennessee and Missouri) demonstrated statistically significant female preference for sympatric males, this trend was observed in all populations tested. Further, we show that (i) males from Tennessee, Missouri, and New Mexico differ in their scent, (ii) females may use population‐specific scents to discriminate among males, and (iii) females whose antennae have been surgically removed are unable to recognize acceptable mates. New Mexico males, which were never accepted by either Tennessee or Missouri females, became acceptable mates when crowded with Tennessee or Missouri males prior to copulation. We infer that male odor may be an important factor in determining cucumber beetle mating success.     

Diapause affects cuticular hydrocarbon composition and mating behavior of both sexes in Drosophila montana

Envipnmental cues,mainly photoperiod and temperature,are known to control female adult reproductive diapause in several insect species.Diapause enhances female survival during adverse conditions and postpones progeny production to the favorable season.Male diapause(a reversible inability to inseminate receptive females)has been studied much less than female diapause.However,if the males maximized their chances to fertilize females while minimizing their energy expenditure,they would be expected to be in diapause at the same time as females.We investigated Drosophila montana male mating bchavior under short-day conditions that induce diapause in females and found the males to be reproductively inactive.We also found that males reared under long-day conditions(reproducing individuals)court reproducing postdiapause fermales,but not diapausing ones.The diapausing fies of both sexes had more long-chain and less short-chain hydrocarbons on their cuticle than the reproducing ones,which presumably increase their survival under stressful conditions,but at the same time decrease their attractiveness.Our study shows that the mating behavior of females and males is well coordinated during and afier overwintering and it also gives support to the dual role of insect cuticular hydrocarbons in adaptation and mate choice.     

Female mate choice and male–male competition in Tonkean macaques: Who decides?

The theory of sexual selection predicts that females should be discriminatory in the choice of sexual partners. Females can express their choice in two ways. In direct mate choice, they show preferences for certain partners. In indirect mate choice, they select partners by displaying sexually attractive traits, thus eliciting contest competition between males. We focused on a primate species in which females advertise the timing of their ovulation and studied the balance between these two choice strategies. We tested predictions related to three hypotheses about direct and indirect female choice, namely the best‐male, graded‐signal and weak‐selectivity hypotheses. We investigated the sexual and agonistic interactions occurring during oestrous periods in five captive groups of Tonkean macaques (Macaca tonkeana). The results showed that dominant males used mate guarding to monopolise sexual access to parous females that were in the fertile stage of their reproductive cycle, while lower‐ranking males monitored only nulliparous females. The distribution of sexual presentations indicated that females accepted different types of partners, supporting the weak‐selectivity hypothesis regarding direct mate choice. The analysis of behavioural sequences revealed that mate‐guarding males used mild coercive behaviours to prevent females from mating with other males at conception time. The distribution of mounts showed that females mainly mated with dominant males, which leads us to argue that the best‐male hypothesis provides the most parsimonious explanation regarding indirect mate choice in Tonkean macaques. At the individual level, it may be concluded that male competitive strategies prevented females from exercising direct mate choice. At the evolutionary level, however, female sexual advertising and thus indirect choice promoted competition between males. The outcome is that indirect mate choice appears more important than direct mate choice in female Tonkean macaques.     

Premating isolation is determined by larval rearing substrates in cactophilic Drosophila mojavensis. IX. Host plant and population specific epicuticular hydrocarbon expression influences mate choice and sexual selection

Sexual signals in cactophilic Drosophila mojavensis include cuticular hydrocarbons (CHCs), contact pheromones that mediate female discrimination of males during courtship. CHCs, along with male courtship songs, cause premating isolation between diverged populations, and are influenced by genotype × environment interactions caused by different host cacti. CHC profiles of mated and unmated adult flies from a Baja California and a mainland Mexico population of D. mojavensis reared on two host cacti were assayed to test the hypothesis that male CHCs mediate within‐population female discrimination of males. In multiple choice courtship trials, mated and unmated males differed in CHC profiles, indicating that females prefer males with particular blends of CHCs. Mated and unmated females significantly differed in CHC profiles as well. Adults in the choice trials had CHC profiles that were significantly different from those in pair‐mated adults from no‐choice trials revealing an influence of sexual selection. Females preferred different male CHC blends in each population, but the influence of host cactus on CHC variation was significant only in the mainland population indicating population‐specific plasticity in CHCs. Different groups of CHCs mediated female choice‐based sexual selection in each population suggesting that geographical and ecological divergence has the potential to promote divergence in mate communication systems.     

Mated Redback Spider Females Re-Advertise Receptivity Months after Mating

In many species, selection acts on males to recognize female reproductive status at a distance using pheromones. Unmated females may actively seek to attract males; however, mated females may become cryptic to avoid attracting additional males if multiple matings are costly. Although females of many species cease pheromone production after mating, it is often unclear whether this is a strategic part of a female reproductive strategy, or whether this is because of chemical manipulation by males. If variation in pheromone production is part of the female’s strategy, then we predicted mated females should eventually re‐advertise receptivity if the benefits of multiple mating increase with time since copulation (e.g. because of sperm depletion). Here, we tested this prediction in Australian redback spiders (Latrodectus hasselti). First, we replicated earlier results by showing that virgin males discriminate female maturity and mating status based exclusively on web‐borne chemicals. Our results show this difference must arise from a change in chemical deposition in the web as we controlled for web volume differences between mated and virgin females. Male activity on extracts from webs of virgin females exceeded activity on a solvent control and on extracts of webs of just‐mated females—confirming that female redbacks cease pheromone production immediately after mating. Second, we tested a new prediction that mated females might re‐advertise receptivity near the end of their normal breeding season to replenish diminished sperm stores prior to overwintering. Consistent with the prediction of strategic advertisement, we show that male activity on extracts from females’ webs increased significantly 3 mo after the female first mated (typical length of the breeding season). Thus, these females had begun to add pheromone to their web again. At this time, 26% of these females re‐mated with a second male. If females re‐advertise receptivity to ensure adequate sperm stores, then we predicted a positive relationship between female reproductive output during the 3‐mo interval after copulation and the subsequent intensity of male response to web extracts. However, differences in male activity time were not related to the total number of spiderlings or the number of egg sacs a female had produced during the 3‐mo interval after the first copulation. This result could arise if male chemical manipulation of female receptivity decreases with time after copulation, or if the testing interval used in our study was too long to reveal variation in sperm depletion in females. Thus, although our results are consistent with the idea that females strategically alter pheromonal advertisement, we cannot distinguish this from the hypothesis that female receptivity arises from chemical manipulation by males.