Does female control and male mating system predict courtship investment and mating outcomes? A comparative study in five widow spider species (genus Latrodectus) tested under similar laboratory conditions.

BACKGROUND: Male courtship investment may evolve in response to the male's expectation of future mating opportunities or the degree of female control during mating interactions. We used a comparative approach to test this hypotheses by assessing the courtship and mating behaviors of five widow spider species (genus Latrodectus) under common laboratory conditions. We predicted male investment in courtship would be higher in species where males mate only once because of high cannibalism rates (monogyny, L. geometricus, L. hasselti, L. mirabilis), compared to species with rare cannibalism (L. mactans, L. hesperus) in which males should reserve energy for future mating opportunities. Increased male investment, measured as courtship duration, might also evolve with increased female control over mating outcomes if females prefer longer courtships. We tested this by assessing the frequency of copulations, timing of sexual cannibalism, and the degree of female-biased size dimorphism, which is expected to be negatively correlated with the energetic cost of rebuffing male mating attempts. RESULTS: Copulation frequency was consistently lower in species with extreme female-skewed size dimorphism, and where sexual cannibalism was more prevalent, suggesting the importance of female control for mating outcomes. We confirmed significant interspecific variation in average courtship duration, but contrary to predictions, it was not predicted by male mating system, and there was no consistent link between courtship duration and sexual size dimorphism. CONCLUSION: We show that the degree of sexual dimorphism is not only correlated with sexual cannibalism, but also with mating success since restriction of male copulation frequency by female Latrodectus affects paternity. However, predictions about male mating system or female control affecting courtship duration were not supported. We propose that the form of female control over mating and cannibalism, and male responses, might be more informative for understanding the evolution of courtship duration. For example, male tactics to avoid female aggression may drive lower courtship duration in species like L. mirabilis. Nonetheless, our results differ from inferences based on published studies of each species in isolation, illuminating the need for standardized data collection for behavioural comparative studies.

The protamines of the noble false widow spider Steatoda nobilis provide an example of liquid-liquid phase separation chromatin transitions during spermiogenesis.

While there is extensive information about sperm nuclear basic proteins (SNBP) in vertebrates, there is very little information about Arthropoda by comparison. This paper aims to contribute to filling this gap by analyzing these proteins in the sperm of the noble false widow spider Steatoda nobilis (Order Araneae, Family Theridiidae). To this end, we have developed a protein extraction method that allows the extraction of cysteine-containing protamines suitable for the preparation and analysis of SNBPs from samples where the amount of starting tissue material is limited. We carried out top-down mass spectrometry sequencing and molecular phylogenetic analyses to characterize the protamines of S. nobilis and other spiders. We also used electron microscopy to analyze the chromatin organization of the sperm, and we found it to exhibit liquid-liquid phase spinodal decomposition during the late stages of spermiogenesis. These studies further our knowledge of the distribution of SNBPs within the animal kingdom and provide additional support for a proposed evolutionary origin of many protamines from a histone H1 (H5) replication-independent precursor.

Pre-copulatory choices drive post-copulatory decisions: mechanisms of female control shift across different life stages.

BACKGROUND: The 'wallflower' hypothesis proposes females mate indiscriminately to avoid reproductive delays. Post-copulatory mechanisms may then allow 'trading up', favouring paternity of future mates. We tested links between pre- and post-copulatory choice in Latrodectus geometricus female spiders paired sequentially with two males. These females copulate as adults or as subadults and store sperm in paired spermathecae. Choosy adults have a higher risk of delays to reproduction than subadults. RESULTS: We predicted low pre-copulatory, but high post-copulatory choice at first matings for adults and the opposite for subadults. At second matings, we expected all females would prefer males superior to their first. We found all females mated indiscriminately at their first pairing, but in contrast to subadults, adults usually allowed only a single insertion (leaving one of their paired spermatheca empty); a mechanism of post-copulatory choosiness. Adult-mated females were more likely to remate than subadult-mated females when they became adults, showing a preference for larger males, while subadult-mated females tended to prefer males of greater size-corrected mass. CONCLUSIONS: Our results show that the 'wallflower' effect and 'trading up' tactics can be utilized at different life stages, allowing females to employ choice even if rejecting males is costly.

Realized niche shift of an invasive widow spider: drivers and impacts of human activities.

BACKGROUND: Predicting invasiveness requires an understanding of the propensity of a given species to thrive in areas with novel ecological challenges. Evaluation of realized niche shift of an invasive species in its invasive range, detecting the main drivers of the realized niche shift, and predicting the potential distribution of the species can provide important information for the management of populations of invasive species and the conservation of biodiversity. The Australian redback spider, Latrodectus hasselti, is a widow spider that is native to Australia and established in Japan, New Zealand, and Southeast Asia. We used ecological niche models and ordinal comparisons in an integrative method to compare the realized niches of native and invasive populations of this spider species. We also assessed the impact of several climatic predictor variables and human activity on this niche shift. We hypothesized that human impact is important for successful establishment of this anthropophilic species, and that climatic predictor variables may determine suitable habitat and thus predict invasive ranges. RESULTS: Our models showed that L. hasselti distributions are positively influenced by human impact in both of the native and invasive ranges. Maximum temperature was the most important climatic variable in predictions of the distribution of native populations, while precipitation seasonality was the most important in predictions of invasive populations. The realized niche of L. hasselti in its invasive range differed from that in its native range, indicating possible realized niche shift. CONCLUSIONS: We infer that a preference for human-disturbed environments may underlie invasion and establishment in this spider species, as anthropogenic habitat modifications could provide shelters from unsuitable climatic conditions and extreme climatic stresses to the spiders. Because Australia and the countries in which the species is invasive have differing climates, differences in the availability of certain climatic conditions could have played a role in the realized niche shift of L. hasselti.

Risky business: males choose more receptive adults over safer subadults in a cannibalistic spider.

Understanding factors affecting male mate choice can be important for tracking the dynamics of sexual selection in nature. Male brown widow spiders (Latrodectus geometricus) mate with adult as well as immature (subadult) females. Mating with adults involves costly courtship with a repertoire of signaling behaviors, and typically ends with cannibalism ("self-sacrifice" initiated by male somersault). Mating with subadults involves brief courtship with behavioral repertoire reduced to one component (vibration) and no cannibalism. We examined male mate choice as a function of risks associated with different types of mates and the cues available to courting males. Previous studies showed male preference for adults based on air-borne pheromones, but it was unclear whether that preference is maintained after males reach female's webs. We show that males prefer adults also based on silk-borne contact cues. To determine which types of cues trigger different courtship components, we swapped adults and subadults between webs. We showed that contact with adult females' webs triggers two courtship behaviors from the repertoire, with adult female's bodies triggering additional behaviors. However, vibrational signals occur regardless of the web origin or female developmental stage. We conclude that males recognize subadult females as potential mates, but are more likely to invest in costly courtship behaviors and mating attempts with adults. In our experiments, subadults were less likely to mate than adults. We conclude that mating with adults could be the preferred option for males because of the higher likelihood of copulation, even at the cost of a higher risk of cannibalism.

Invasive Widow Spiders Perform Differently at Low Temperatures from Conspecifics from the Native Range.

Temperature challenges are one of the leading abiotic causes of success or failure of non-native species in a novel environment, and this is particularly true for low temperatures. Establishing and reproducing in a novel thermal environment can alter survival, behavior, and traits related to fitness. It has been proposed that plasticity or adaptation of thermal tolerance may allow an introduced species to thrive, or that successful invaders may be those with a thermal breadth in their native habitat that encompasses their new environment. Here, we tested these hypotheses using native and invasive populations of Australian redback spiders (Latrodectus hasselti). We measured how exposure to temperatures common to invasive and native range habitats (exposure to 15 and 25°C, respectively) affected behavioral and life-history traits and trade-offs that may underlie fitness in an invasive population detected in 1995 in Japan and a native population from Australia. We found that the critical thermal minimum (CTmin) was higher in the invasive population from Japan than in the native population, but critical thermal maximum (CTmax) did not differ between populations. Compared to the invasive population, eggs from the native population had a longer development time and lower hatching success at 15°C. Both populations performed equally well at 25°C, as measured by egg development time and hatching success. Invasive juveniles tested at 15°C were faster to explore a novel environment and bolder compared to those tested at 25°C. In comparison, the native population showed faster average exploration, with no differences in boldness or exploration at the two development or testing temperatures. Overall, L. hasselti from Japan maintained hatching success and development across a wider temperature range than the native population, indicating greater thermal breadth and higher behavioral plasticity. These results support the importance of plasticity in thermal tolerance and behavior for a successful invasion under novel environmental temperatures.

Juvenile Experience with Male Cues Triggers Cryptic Choice Mechanisms in Adult Female Redback Spiders.

Female choice may be linked to population density if the expected encounter rates with potential mates affects choosiness (the energy and risk engaged to express mate preferences). Choosiness should covary with male availability, which could be assessed using the social cues available during development. We tested whether the exposure of juvenile females to cues of male density affected the mechanisms of choosiness of adult Latrodectus hasselti spiders in two experiments simulating natural contexts. The juvenile females were exposed to (1) volatile chemicals from two densities of adult males (airborne cues), and (2) tactile, vibrational and chemical cues from adult males or other females (cohabitation cues). As adults, the females mated readily, regardless of the treatment, but there was strong evidence for post-copulatory mechanisms of choosiness in females exposed to cues of high male availability. These included abbreviated matings (in both experiments), cannibalism of the males before the mating was complete (cohabitation), and, remarkably, a reduction in the successful placement of internal sperm plugs (cohabitation). These shifts decrease the likelihood that the first mate would monopolize paternity if the female chose to mate again. We conclude that female choosiness may impose a strong selection on males despite the high mating rates, and these effects can hinge on the cues of male availability detected by juveniles.

Female control of a novel form of cannibalism during copulation in a South American widow spider.

Sexual cannibalism is an extreme form of sexual conflict that may have broad evolutionary effects on mating behaviour. Latrodectus spiders (∼30 "widow" species) could enable comparative tests of the evolution of sexual cannibalism, but most species are poorly-studied. Here we describe the mating and remating behaviour of a species endemic to South America, L. mirabilis, with a focus on the occurrence and consequences of sexual cannibalism. Mating and cannibalistic behaviours were novel relative to other Latrodectus species. Cannibalism started during copulation when females grasped the male's legs with their chelicerae, and continued as females pulled the male's abdomen onto their chelicerae, without interrupting copulation. Cannibalism was initiated by females, and not facilitated by males as was observed in other Latrodectus species. Females frequently remated however, so cannibalized males might lose paternity to future rivals. We report high rates of cannibalism, with 70 % of males killed by females during their first mating, and 85 % killed by previously-mated females. We discuss our novel findings in the context of previous observations in other Latrodectus species, proposing that foundational studies in a wider range of species are necessary to support comparative tests about the evolution of sexual cannibalism within this model taxon.

Black widows as plastic wallflowers: female choosiness increases with indicators of high mate availability in a natural population.

Female choice is an important driver of sexual selection, but can be costly, particularly when choosy females risk remaining unmated or experience delays to reproduction. Thus, females should reduce choosiness when mate encounter rates are low. We asked whether choosiness is affected by social context, which may provide reliable information about the local availability of mates. This has been demonstrated in the lab, but rarely under natural conditions. We studied western black widow spiders (Latrodectus hesperus) in the field, placing experimental final-instar immature females so they were either 'isolated' or 'clustered' near naturally occurring conspecifics (≥10 m or ≤1 m, respectively, from a microhabitat occupied by at least one other female). Upon maturity, females in both treatments were visited by similar numbers of males, but clustered females were visited by males earlier and in more rapid succession than isolated females, confirming that proximity to conspecifics reduces the risk of remaining unmated. As predicted, isolated females were less choosy in staged mating trials, neither rejecting males nor engaging in pre-copulatory cannibalism, in contrast to clustered females. These results demonstrate that exposure of females to natural variation in demography in the field can alter choosiness of adults. Thus, female behaviour in response to cues of local population density can affect the intensity of sexual selection on males in the wild.

Immature mating as a tactic of polygynous male western widow spiders.

In polygynous species, males typically mate with more than one female, and male investment in any one mating should decrease if securing that mating reduces future mating opportunities. In contrast, monogynous males mate only once, so they should invest maximally in that single mating. In two monogynous, sexually cannibalistic Latrodectus spider species (L. geometricus, L. hasselti), males can mate and fertilize immature females. This recently described tactic is time-consuming because males must wait days for females to be capable of mating. It is also risky since immature females approached too early may kill males outright. However, if males typically find only one female in their lifetime, increasing the opportunity to mate may be worth these costs. We investigated whether this tactic is also practiced by a polygynous congener, L. hesperus, in which males typically mate more than once and may avoid time-consuming, risky encounters. In laboratory trials, we showed that males copulate with immature females. Moreover, males mounted immatures more rapidly, copulated for longer, and fathered more offspring than males that mated adults females. We concluded that monogyny is not a necessary condition for immature mating to be favored as an alternative reproductive tactic and suggest that it may be common in other spider taxa.

Male black widows parasitize mate-searching effort of rivals to find females faster.

Mate-searching success is a critical precursor to mating, but there is a dearth of research on traits and tactics that confer a competitive advantage in finding potential mates. Theory and available empirical evidence suggest that males locate mates using mate-attraction signals produced by receptive females (personal information) and avoid inadvertently produced cues from rival males (social information) that indicate a female has probably already mated. Here, we show that western black widow males use both kinds of information to find females efficiently, parasitizing the searching effort of rivals in a way that guarantees competition over mating after reaching a female's web. This tactic may be adaptive because female receptivity is transient, and we show that (i) mate searching is risky (88% mortality) and (ii) a strongly male-biased operational sex ratio (from 1.2 : 1 to more than 10 : 1) makes competition inevitable. Males with access to rivals' silk trails moved at higher speeds than those with only personal information, and located females even when personal information was unreliable or absent. We show that following rivals can increase the potential for sexual selection on females as well as males and argue it may be more widespread in nature than is currently realized.

Natural Compounds as Spider Repellents: Fact or Myth?

Although some spiders are globally invasive, found at high densities, and may be considered pests (particularly those that are toxic to humans), there are few pest management methods based on experimental data. 'Common wisdom' and advertisements on internet websites assert that a number of natural substances repel spiders. We tested whether the three substances cited most frequently (lemon oil, peppermint oil, and chestnut-fruits) effectively repelled female spiders or whether these were myths. We presented each of the putative repellents versus a control in a two-choice assay and tested responses of females of three invasive spider species in two different families: theridiids, Latrodectus geometricus C. L. Koch (Araneae: Theridiidae) and Steatoda grossa C. L. Koch (Araneae: Theridiidae) and the araneid, Araneus diadematus Clerck . Chestnuts (Araneae: Araneidae) and mint oil strongly repelled L. geometricus and A. diadematus. S. grossa was less sensitive to these chemicals but had a slight tendency to avoid chestnuts. However, lemon oil, the substance most likely to be cited as a repellent (over 1,000,000 hits on Google), had no effect on any of these spiders. We conclude that volatiles released by mint oil and chestnuts may be effective in deterring spider settlement in two different families of spiders, but lemon oil as a repellent is a myth.

Neutral fitness outcomes contradict inferences of sexual 'coercion' derived from male's damaging mating tactic in a widow spider.

Sexual conflict over mating frequency has driven the evolution of morphological and behavioural traits across taxa. Interactions may be termed 'coercive' and assumed to arise from conflict when male mating behaviours cause physical injury to females and females appear to resist injurious matings.However, coercion per se occurs only if the behaviour reduces female fitness; and such outcomes are rarely measured. Here we show that a damaging mating tactic, apparently adaptive for males, is not coercive for females. Adult male Latrodectus spiders mate with immature females after tearing the exoskeleton covering the female's recently-developed reproductive tract, which can cause haemolymph bleeding. We show that, relative to pairings with adult females, males use reduced courtship displays when approaching immature females, which in some cases respond with elevated deterrent behavioural responses. Nevertheless, we found no reproductive cost for immature-mated females in terms of longevity, fertility or fecundity. Moreover, most immature-mated females did not produce sex pheromones as adults, so did not seek additional matings. Thus, despite the appearance of conflict there is no evidence that immature-mating is coercive. These results show it is critical to measure fitness outcomes, in addition to behavioural responses, to test for coercion.

Copulation with immature females increases male fitness in cannibalistic widow spiders.

Copulatory cannibalism of male 'widow' spiders (genus Latrodectus) is a model example of the extreme effects of sexual selection, particularly in L. hasselti and L. geometricus where males typically facilitate cannibalism by females and mate only once. We show that these males can increase their reproductive success by copulating with final-instar, immature females after piercing the female's exoskeleton to access her newly developed sperm storage organs. Females retain sperm through their final moult and have similar fecundity to adult-mated females. This is an adaptive male tactic because immature mating increases insemination success relative to adult mating (which predicts higher paternity) and moreover, rarely ends in cannibalism, so males can mate again. Although successful only during a brief period before the female's final moult, males may employ this tactic when they associate with final-instar females in nature. Consistent with this, one-third of L. hasselti females collected as immatures in nature were already mated. Immature mating alters sexual selection on these otherwise monogynous males, and may explain male traits allowing facultative polygyny in Latrodectus Since male cohabitation with immature females is common among invertebrates, immature mating may be a widespread, previously unrecognized mating tactic, particularly when unmated females are of high reproductive value.

Metabolic efficiency in courtship favors males with intermediate mass in the Australian redback spider, Latrodectus hasselti.

Recent studies have suggested that metabolic efficiency may be an important factor in male mating success when females require vigorous and/or prolonged courtship. In capital breeding animals in which a male's resource pool is fixed at adulthood the relationship between energy expenditure and courtship performance may be especially important, as males are expected to utilize their finite resources efficiently when soliciting mates. Males may benefit from being efficient, i.e., achieving a sufficiently high level of courtship signaling at low energetic cost, if it enables them to acquire mates before their limited energy reserves are depleted. We investigated the relationship between metabolic efficiency and courtship vibrational signaling in the Australian redback spider, Latrodectus hasselti, a semelparous capital breeder where males invest heavily in courtship to secure a mating. We assessed metabolic rate in a sample of males and measured two courtship components (duty cycle and amplitude) that reflected the energy content of web-borne vibrations. We then calculated two indices of metabolic efficiency for these courtship properties. There was a quadratic relationship between mass and duty cycle such that the highest duty cycle signals were performed by males having intermediate mass. Furthermore, intermediate-mass males were also the most metabolically efficient. Prolonged courtship is necessary in L. hasselti for successful mating, and the results of this study suggest that intermediate-mass males are superior courters because they utilize their finite resource pool most efficiently to produce high energy vibrational signals.

Sibling cannibalism in a web-building spider: effects of density and shared environment.

Sibling cannibalism occurs across diverse taxa and can affect population size and structure, as well as the fitness of parents and the cannibal, via density effects and variation in individual propensity to cannibalize. We examined these effects on sibling cannibalism in juveniles of a web-building spider (Latrodectus hasselti, Australian redbacks). Adult redbacks are solitary, but juveniles live in clusters of variable density for a week after hatching. We confined newly hatched siblings from a singly-mated female to a low or high density treatment in a split-clutch design, then left spiderlings unfed for a week. Our results showed no effect of density on overall cannibalism levels, but a strong correlation between cannibalism counts from the same maternal lines across densities. Unlike web-bound sit-and-wait predators, wandering spiders that are active hunters have been shown to experience density-dependent cannibalism. In contrast, we suggest sibling cannibalism in web-building spiders may be density independent because early cohabitation on the web selects for elevated tolerance of conspecifics. We conclude that, rather than being linked to density, cannibalism of siblings in these species may be controlled more strongly by variation in individual propensity to cannibalize.

Mating system does not predict permanent sperm depletion in black widow spiders.

Variation in sperm production is strongly influenced by mating system across taxa. Recent work in spiders suggests that males of some species show termination of spermatogenesis before their adult molt and thus an inability to produce sperm after maturation. This permanent sperm depletion (PSD) has been hypothesized to co-occur with monogyny, genital mutilation, or sexual cannibalism because the maintenance of continual sperm supplies is not necessary for species where males can expect only one mating opportunity. Here we test this hypothesis in two congeners exhibiting genital mutilation: the sexually cannibalistic, monogynous Australian redback spider Latrodectus hasselti and the polygynous Western black widow Latrodectus hesperus. We report that PSD does not occur in adult males of either species, and show that males transfer sperm into their copulatory organs multiple times as adults. These data suggest evolutionary links between mating system and investment in sperm production may be more complex than currently appreciated.

Developmental plasticity in metabolic rates reinforces morphological plasticity in response to social cues of sexual selection.

Developmental plasticity allows individuals to match their phenotype to the competitive environment they are most likely to encounter. Although there are numerous studies that demonstrate adaptive shifts in life-history and metric traits, we still have a poor understanding of whether internal physiological processes demonstrate plasticity and whether this plasticity supports life-history and metric traits. Here we use the Australian redback spider (Latrodectus hasselti), a species that demonstrates adaptive developmental shifts in response to the availability of females and the density of males, to examine the relationship between the routine metabolic rate (RMR) and the expression of size, body-condition and development rate. We reared immature males in three diet treatments, and in social environments that varied the presence/absence of females and the density of males and measured their RMR, weight, size and developmental rate at maturity. We show that although RMR decreased with decreasing resource abundance, RMR was positively correlated with the density of rivals. Moreover, RMR was not correlated with size or body-condition at maturity. Our results demonstrate that plasticity in the RMR supports plasticity in metric and life-history traits to create an integrated phenotype that matches the competitive environment.

Examination of prior contest experience and the retention of winner and loser effects.

In many animal taxa, prior contest experience affects future performance such that winning increases the chances of winning in the future (winner effect) and losing increases the chances of losing in the future (loser effect). It is, however, not clear whether this pattern typically arises from experience effects on actual or perceived fighting ability (or both). In this study, we looked at winner and loser effects in the jumping spider Phidippus clarus. We assigned winning or losing experience to spiders and tested them against opponents of similar fighting ability in subsequent contests at 1-, 2-, 5-, and 24-h intervals. We examined the strength of winner and loser effects, how long effects persist, as well as how experience affected perceived and actual fighting ability. Our results demonstrate that winner and loser effects are of approximately the same magnitude, although loser effects last longer than winner effects. Our results also demonstrate that previous experience alters actual fighting ability because both the assessment and escalation periods were affected by experience. We suggest that the retention time of experience effects depends on expected encounter rates as well as other behavioral and ecological factors. In systems with short breeding seasons and/or rapidly fluctuating populations, context-dependent retention of experience effects may allow males to track their status relative to the fluctuating fighting ability of local competitors without paying the costs necessary to recall or assess individual competitors.