Alternative Life-history Strategies Research Articles

Life history correlations and trade-offs resulting from selection for dispersal in Tribolium castaneum.

Dispersal is an important facet of the life history of many organisms and is therefore subject to selective pressure, but does not evolve in isolation. Across nature there are examples of dispersal syndromes, life history strategies in which suites of traits coevolve and covary with dispersal in combinations that serve to maximise fitness in a given ecological context. The red rust flour beetle, Tribolium castaneum, is a model organism and globally significant post-harvest pest which relies on dispersal to reach new patches of ephemeral habitat. Dispersal behaviour in Tribolium has a strong genetic basis. However, a robust understanding of the relationship between dispersal and other life history components, which could elucidate evolutionary processes and allow pest managers to control their spread and reduce the impact of infestation, is currently lacking. Here we use highly replicated lines of T. castaneum previously artificially selected for divergent small-scale dispersal propensity, to robustly test several important life history components: reproductive strategy, development time and longevity. As predicted, we find that a suite of important change as result of our selection on dispersal; high dispersal propensity is associated with a lower number of longer mating attempts by males, lower investment in early-life reproduction by females, slower development of later-laid offspring and longer female lifespan. These findings indicate that correlated intraspecific variation in dispersal and related traits may represent alternative life history strategies in T. castaneum. We therefore suggest that pest management efforts to mitigate the species' agro-economic impact should consider the eco-evolutionary dynamics within multiple life-histories. The benefits of doing so could be felt both through improved targetting of efforts to reduce spread, and also in forecasting of how the selection pressures applied through pest management are likely to affect pest evolution.

Alternative life-history strategy contributions to effective population size in a naturally spawning salmon population.

Alternative life-history tactics are predicted to affect within-population genetic processes but have received little attention. For example, the impact of precocious males on effective population size (N e) has not been quantified directly in Pacific salmon Oncorhynchus spp., even though they can make up a large percentage of the total male spawners. We investigated the contribution of precocial males ("jacks") to N e in a naturally spawning population of Coho Salmon O. kisutch from the Auke Creek watershed in Juneau, Alaska. Mature adults that returned from 2009 to 2019 (~8000 individuals) were genotyped at 259 single-nucleotide polymorphism (SNP) loci for parentage analysis. We used demographic and genetic methods to estimate the effective number of breeders per year (N b). Jack contribution to N b was assessed by comparing values of N b calculated with and without jacks and their offspring. Over a range of N b values (108-406), the average jack contribution to N b from 2009 to 2015 was 12.9% (SE = 3.8%). Jacks consistently made up over 20% of the total male spawners. The presence of jacks did not seem to influence N b/N. The linkage disequilibrium N e estimate was lower than the demographic estimate, possibly due to immigration effects on population genetic processes: based on external marks and parentage data, we estimated that immigrant spawners produced 4.5% of all returning offspring. Our results demonstrate that jacks can influence N b and N e and can make a substantial contribution to population dynamics and conservation of threatened stocks.

Evidence for a single, ancient origin of a genus-wide alternative life history strategy

Understanding the evolutionary origins and factors maintaining alternative life history strategies (ALHS) within species is a major goal of evolutionary research. While alternative alleles causing discrete ALHS are expected to purge or fix over time, one-third of the ~90 species of Colias butterflies are polymorphic for a female-limited ALHS called Alba. Whether Alba arose once, evolved in parallel, or has been exchanged among taxa is currently unknown. Using comparative genome-wide association study (GWAS) and population genomic analyses, we placed the genetic basis of Alba in time-calibrated phylogenomic framework, revealing that Alba evolved once near the base of the genus and has been subsequently maintained via introgression and balancing selection. CRISPR-Cas9 mutagenesis was then used to verify a putative cis-regulatory region of Alba, which we identified using phylogenetic foot printing. We hypothesize that this cis-regulatory region acts as a modular enhancer for the induction of the Alba ALHS, which has likely facilitated its long evolutionary persistence.

Local adaptation of seed and seedling traits along a natural aridity gradient may both predict and constrain adaptive responses to climate change.

Variation in seed and seedling traits underlies how plants interact with their environment during establishment, a crucial life history stage. We quantified genetic-based variation in seed and seedling traits in populations of the annual plant Plantago patagonica across a natural aridity gradient, leveraging natural intraspecific variation to predict how populations might evolve in response to increasing aridity associated with climate change in the Southwestern U.S. We quantified seed size, seed size variation, germination timing, and specific leaf area in a greenhouse common garden, and related these traits to the climates of source populations. We then conducted a terminal drought experiment to determine which traits were most predictive of survival under early-season drought. All traits showed evidence of clinal variation-seed size decreased, germination timing accelerated, and specific leaf area increased with increasing aridity. Populations with more variable historical precipitation regimes showed greater variation in seed size, suggestive of past selection shaping a diversified bet-hedging strategy mediated by seed size. Seedling height, achieved via larger seeds or earlier germination, was a significant predictor of survival under drought. We documented substantial interspecific trait variation as well as clinal variation in several important seed and seedling traits, yet these slopes were often opposite to predictions for how individual traits might confer drought tolerance. This work shows that plant populations may adapt to increasing aridity via correlated trait responses associated with alternative life history strategies, but that trade-offs might constrain adaptive responses in individual traits.

Metabolite Changes in Orange Dead Leaf Butterfly Kallima inachus during Ontogeny and Diapause.

Holometabolism is a form of insect development which includes four life stages: egg, larva, pupa, and imago (or adult). The developmental change of whole body in metabolite levels of holometabolous insects are usually ignored and lack study. Diapause is an alternative life-history strategy that can occur during the egg, larval, pupal, and adult stages in holometabolous insects. Kallima inachus (Lepidoptera: Nymphalidae) is a holometabolous and adult diapausing butterfly. This study was intended to analyze metabolic changes in K. inachus during ontogeny and diapause through a non-targeted UPLC-MS/MS (ultra-performance liquid chromatograph coupled with tandem mass spectrometry) based metabolomics analysis. A variety of glycerophospholipids (11), amino acid and its derivatives (16), and fatty acyls (nine) are crucial to the stage development of K. inachus. 2-Keto-6-acetamidocaproate, N-phenylacetylglycine, Cinnabarinic acid, 2-(Formylamino) benzoic acid, L-histidine, L-glutamate, and L-glutamine play a potentially important role in transition of successive stages (larva to pupa and pupa to adult). We observed adjustments associated with active metabolism, including an accumulation of glycerophospholipids and carbohydrates and a degradation of lipids, as well as amino acid and its derivatives shifts, suggesting significantly changed in energy utilization and management when entering into adult diapause. Alpha-linolenic acid metabolism and ferroptosis were first found to be associated with diapause in adults through pathway analyses. Our study lays the foundation for a systematic study of the developmental mechanism of holometabolous insects and metabolic basis of adult diapause in butterflies.

Coexistence of two newt species in a transition zone of range overlap

Abstract Theory suggests that spatial segregation of similar, co-occurring species may be driven by alternative innate life history and dispersal strategies, and that it operates through catastrophic events. An inventory of the evolutionary closely related small-bodied newts Lissotriton helveticus and L. vulgaris in the northwest of France demonstrated the species’ spatial partitioning, with L. vulgaris dominating in two pond-rich and historically disturbed coastal areas and L. helveticus prevailing inland where ponds are sparser. Population numbers were followed over several decades (1975–2021) in a pond within the narrow (ca. 2000 m wide) species transition zone. Early in the temporal survey (1986) a massive die-off was observed of two-third of the L. helveticus breeding population from a late frost event. Yet, the contribution of L. helveticus to the newt assemblage was more or less stable around 60%, even though the total population size fluctuated by an order of magnitude. Lissotriton vulgaris and a third species, Ichthyosaura alpestris, made up ca. 30% and 10% of the total till 1993, after which date their relative contributions reversed. These data suggest that a state shift may have occurred among the latter two species and that the assumed two-species dynamics of Lissotriton underlying the study has been an oversimplification. The local decline of L. vulgaris is paralleled by the loss of well-vegetated ponds from the wider agricultural terrain that affects this species more than L. helveticus and I. alpestris.

Streblospio benedicti: A genetic model for understanding the evolution of development and life-history.

Investigating developmental evolution usually requires comparing differences across related species to infer how phenotypic change results from embryological modifications. However, when comparing organisms from different environments, ecologies, and evolutionary histories there can be many confounding factors to finding a genetic basis for developmental differences. In the marine annelid Streblospio benedicti, there are two distinct types of offspring with independent developmental pathways that converge on the same adult phenotype. To my knowledge, S. benedicti is the only known species that has heritable (additive) genetic variation in developmental traits that results in alternative life-history strategies. Females produce either hundreds of small, swimming and feeding larvae, or dozens of large, nonfeeding larvae. The larvae differ in their morphology, ecology, and dispersal potential. This developmental dimorphism makes S. benedicti a unique and useful model for understanding how genetic changes result in developmental modifications that ultimately lead to overall life-history differences. Because the offspring phenotypes of S. benedicti are heritable, we can use forward genetics within a single evolutionary lineage to disentangle how development evolves, and which genes and regulatory mechanisms are involved.

Rhesus macaques compensate for reproductive delay following ecological adversity early in life.

Adversity early in life can shape the reproductive potential of individuals through negative effects on health and life span. However, long‐lived populations with multiple reproductive events may present alternative life history strategies to optimize reproductive schedules and compensate for shorter life spans. Here, we quantify the effects of major hurricanes and density dependence as sources of early‐life ecological adversity on Cayo Santiago rhesus macaque female reproduction and decompose their effects onto the mean age‐specific fertility, reproductive pace, and lifetime reproductive success (LRS). Females experiencing major hurricanes exhibit a delayed reproductive debut but maintain the pace of reproduction past debut and show a higher mean fertility during prime reproductive ages, relative to unaffected females. Increasing density at birth is associated to a decrease in mean fertility and reproductive pace, but such association is absent at intermediate densities. When combined, our study reveals that hurricanes early in life predict a delay‐overshoot pattern in mean age‐specific fertility that supports the maintenance of LRS. In contrast to predictive adaptive response models of accelerated reproduction, this long‐lived population presents a novel reproductive strategy where females who experience major natural disasters early in life ultimately overcome their initial reproductive penalty with no major negative fitness outcomes. Density presents a more complex relation with reproduction that suggests females experiencing a population regulated at intermediate densities early in life will escape density dependence and show optimized reproductive schedules. Our results support hypotheses about life history trade‐offs in which adversity‐affected females ensure their future reproductive potential by allocating more energy to growth or maintenance processes at younger adult ages.

A migratory sparrow has personality in winter that is independent of other traits

Small birds in winter face trade-offs between predation risk and foraging, and alternate life-history strategies may arise from these trade-offs. Animal personality shows similarities with alternative life-history strategies, and using a life-history context to understand personality can provide valuable insights. Golden-crowned sparrows, Zonotrichia atricapilla , a small migratory bird, have a complex winter social system with high site-fidelity, long-term social associations between individuals and competition mediated by badges of status. We asked whether golden-crowned sparrows show personalities during winter, whether these personalities were consistent over 3 years and whether they correlated with social and morphological traits. We found that golden-crowned sparrows had highly repeatable behaviours, as measured in captive behavioural assessments, constituting personalities consistent within one season and over time for up to 3 years, a time span that covers the average life span of the sparrows. While long-term repeatability was present, it varied considerably among different behaviours and time spans, and length of time between measures did not predict the magnitude of repeatability. Two movement-based aspects of sparrow personality were independent of the traits we measured (dominance, badges of status, size and age). However, nonsong vocalizations (a repeatable aspect of personality) correlated with wing length and gold badge size in some years. Because personality did not strongly link to the social traits we measured, sparrow personality could represent a separate axis of variation that might connect with other winter life-history parameters such as foraging, predation response or survival. • We studied whether wintering golden-crowned sparrows show personalities. • Personality was stable both within and across years. • Across-year repeatability showed considerable variability, indicating plasticity. • Personality was independent of dominance, badges of status, size, sex and age. • Personality may represent a separate axis of variation (e.g. survival, foraging).

Offshore behavioral contingent of an estuarine fish population, common snook Centropomus undecimalis

Research that identifies behavioral contingents, portions of a population that exhibit alternate life history strategies or habitat preferences, can provide a better understanding of a species’ resilience to disturbances, changes in environmental factors, and harvest. Sightings of the estuarine-dependent common snook Centropomus undecimalis at offshore reef areas throughout the year in southeast Florida prompted an investigation to determine whether a contingent of the snook population remains offshore year-round and if they can contribute to the inshore population. This study employed underwater visual observations, specimen collections, and acoustic telemetry over 7 yr to document and describe the snook that utilize offshore habitat. Fish were observed in groups of up to 225 individuals, 20.4 km from an inlet, and in waters up to 36.6 m deep. Snook were present in 79.4% of dive surveys conducted on artificial reefs but only in 18.4% of surveys on natural reefs. Acoustic telemetry showed that many fish remained offshore for multiple years. Most sampled fish (89.1%) were spawning capable, with some classified as actively spawning (15.8%). Reports of snook using offshore reefs occur elsewhere, including southwest Florida and the Florida Keys, indicating that this contingent behavior may not be unique to southeast Florida. Evaluating these occurrences and identifying potential triggers that prompt snook to leave the estuary for an offshore environment can aid in determining how the offshore contingent affects the overall population.

Cryptic alternative male mating strategies in invasive alien round goby (Neogobius melanostomus) of the upper Danube River

Male alternative mating strategies are a widespread life history choice, yet they have hardly been assessed in the context of fish invasions. As established proxies for alternative life-history strategies, body length and sexual maturity of Ponto–Caspian round goby (Neogobius melanostomus) from the recently invaded upper Danube River were contrasted and differential trophic niche occupation was assessed. Only about 4% of analyzed gobies qualified as potential sneakers which may be explained by the recent and ongoing invasion process. This study provides evidence for the occurrence of sneaking behavior of invasive round goby in the upper Danube River.

A transposable element insertion is associated with an\xa0alternative life history strategy

Tradeoffs affect resource allocation during development and result in fitness consequences that drive the evolution of life history strategies. Yet despite their importance, we know little about the mechanisms underlying life history tradeoffs. Many species of Colias butterflies exhibit an alternative life history strategy (ALHS) where females divert resources from wing pigment synthesis to reproductive and somatic development. Due to this reallocation, a wing color polymorphism is associated with the ALHS: either yellow/orange or white. Here we map the locus associated with this ALHS in Colias crocea to a transposable element insertion located downstream of the Colias homolog of BarH-1, a homeobox transcription factor. Using CRISPR/Cas9 gene editing, antibody staining, and electron microscopy we find white-specific expression of BarH-1 suppresses the formation of pigment granules in wing scales and gives rise to white wing color. Lipid and transcriptome analyses reveal physiological differences associated with the ALHS. Together, these findings characterize a mechanism for a female-limited ALHS.

Genome evolution and host-microbiome shifts correspond with intraspecific niche divergence within harmful algal bloom-forming Microcystis aeruginosa.

Intraspecific niche divergence is an important driver of species range, population abundance and impacts on ecosystem functions. Genetic changes are the primary focus when studying intraspecific divergence; however, the role of ecological interactions, particularly host-microbiome symbioses, is receiving increased attention. The relative importance of these evolutionary and ecological mechanisms has seen only limited evaluation. To address this question, we used Microcystis aeruginosa, the globally distributed cyanobacterium that dominates freshwater harmful algal blooms. These blooms have been increasing in occurrence and intensity worldwide, causing major economic and ecological damages. We evaluated 46 isolates of M.aeruginosa and their microbiomes, collected from 14 lakes in Michigan, USA, that vary over 20-fold in phosphorus levels, the primary limiting nutrient in freshwater systems. Genomes of M.aeruginosa diverged along this phosphorus gradient in genomic architecture and protein functions. Fitness in low-phosphorus lakes corresponded with additional shifts within M.aeruginosa including genome-wide reductions in nitrogen use, an expansion of phosphorus assimilation genes and an alternative life history strategy of nonclonal colony formation. In addition to host shifts, despite culturing in common-garden conditions, host-microbiomes diverged along the gradient in taxonomy, but converged in function with evidence of metabolic interdependence between the host and its microbiome. Divergence corresponded with a physiological trade-off between fitness in low-phosphorus environments and growth rate in phosphorus-rich conditions. Co-occurrence of genotypes adapted to different nutrient environments in phosphorus-rich lakes may have critical implications for understanding how M.aeruginosa blooms persist after initial nutrient depletion. Ultimately, we demonstrate that the intertwined effects of genome evolution, host life history strategy and ecological interactions between a host and its microbiome correspond with an intraspecific niche shift with important implications for whole ecosystem function.

Behaviourally specialized foragers are less efficient and live shorter lives than generalists in wasp colonies

A widely held assumption in ecology is that specialists are more efficient than generalists. However, empirical evidence for this fundamental assumption is surprisingly scarce and often contradictory. Theoretically, the evolution of alternative life history strategies is underpinned by a trade-off between activity levels and survival. We investigated the consequences of specialization in a foraging context, by comparing the performance and longevity of closely related individuals in a social insect, the common wasp (Vespula vulgaris). Using radio-frequency identification technology, we monitored the lifetime foraging activity of individual wasps from three colonies kept under natural foraging conditions. Returning foragers were video-recorded as they passed the nest entrance so that their foraging load could be assessed. There were substantial differences in foraging activity and survival within and between colonies. At the colony level, foraging specialization was weak. Yet, workers within each nest demonstrated a remarkable range of foraging specialization levels (defined as the degree of overlap between individual and colony-level task allocation) and efficiencies (defined by the number of successful trips and trip duration). We found that specialist foragers were less efficient than generalist siblings within the same colony. Behavioural specialists accomplished fewer successful trips per foraging day, and their trips were typically relatively longer. Specialized foragers also showed reduced life expectancy. The mortality risk was higher for individuals spending relatively more time in the field, yet we found no link between the level of specialization and relative field exposure. Our extensive dataset of unprecedented detail provides strong empirical evidence that behavioural specialization is not associated with a better lifetime performance, on the contrary, the opposite appears true for the common wasp. We also show that the survival of genetically similar individuals can be linked to life-long differences in behaviour according to classical life-history theory predictions.

Morph- and sex-specific effects of challenging conditions on maintenance parameters in the Gouldian finch.

Intraspecific discrete polymorphism is associated with the use of alternative life-history strategies, reflected by distinct reproductive or copying strategies in individuals of different morphs. Yet, morph-specific costs and benefits related to different life-history strategies remain unclear. Here, we examined in the polymorphic Gouldian finch (Erythrura gouldiae) whether markers of somatic maintenance (body mass, oxidative status and telomere length) differed between red- and black-headed birds under energetically demanding conditions (during heatwaves of different intensity, and during moult or breeding following heatwaves). During heatwaves, red-headed birds showed a homogeneous response, as males and females initially tended to gain mass and had reduced plasma hydroperoxide levels (a marker of oxidative damage) irrespective of heatwave intensity. In contrast, black-headed birds showed a stronger and more heterogeneous response, as black-headed males gained mass at the beginning of the thermoneutral heatwave and showed stable oxidative status, whereas black-headed females lost mass and tended to show higher hydroperoxide levels at the end of the thermocritical heatwave. Following heatwaves, we found morph-specific oxidative costs owing to moult or reproduction, with oxidative markers varying only in black-headed birds. Again, oxidative markers varied differently in black-headed males and females, as plasma antioxidant capacity decreased in moulting or breeding females, whereas males showed higher hydroperoxide levels with larger broods. For the first time, our study highlights that within polymorphic species, some individuals appear more vulnerable than others when coping with energetically demanding conditions. In the context of climate change, such differential effects may ultimately alter the currently observed balance between morphs and sexes within natural populations.

Alternate migration strategies of eastern monarch butterflies revealed by stable isotopes

Abstract Alternative life history strategies are mechanisms by which organisms are able to maximize fitness across a range of environmental conditions. Fitness is maximized by different strategies depending on context, resulting in trade-offs between life history strategies. Monarch butterflies (Danaus plexippus) employ both migratory and resident life history strategies. Since residents breed throughout the year, but migrants overwinter in reproductive diapause, there are fitness trade-offs between the two strategies. We used stable isotope analysis to evaluate the geographic origins of monarchs in a yearround population in south Florida. Based on stable isotope profiles of hydrogen and carbon (δ2H and δ13C values), we found that 48% (16/33) of monarchs collected in south Florida are migrants that originated from outside the sampling region. Migrants had a larger wing length than residents; thus, switching to a resident strategy could alter their probability of reproductive success. Further work is needed to investigate the mechanism underlying this pattern, but these findings show that alternate life history strategies and sex-specific behaviors are underexplored factors influencing monarch migration and evolution.

Implications for introgression: has selection for fast growth altered the size threshold for precocious male maturation in domesticated Atlantic salmon?

BackgroundMature male parr (MMP) represent an important alternative life-history strategy in Atlantic salmon populations. Previous studies indicate that the maturation size threshold for male parr varies among wild populations and is influenced by individual growth, environmental conditions, and genetics. More than ten generations of breeding have resulted in domesticated salmon displaying many genetic differences to wild salmon, including greatly increased growth rates. This may have resulted in domesticated fish with the potential to outgrow the size threshold for early maturation, or evolution of the size threshold of the trait itself. To investigate this, we performed a common-garden experiment under farming conditions using 4680 salmon from 39 families representing four wild, two wild-domesticated hybrid, and two domesticated strains.ResultsDomesticated salmon outgrew wild salmon 2–5-fold, and hybrids displayed intermediate growth. Overall, the numbers of MMP varied greatly among families and strains: averaging 4–12% in domesticated, 18–25% in hybrid, and 43–74% in the wild populations. However, when the influence of growth was accounted for, by dividing fish into lower and upper size modes, no difference in the incidence of MMP was detected among domesticated and wild strains in either size mode. In the lower size mode, hybrids displayed significantly lower incidences of mature males than their wild parental strains. No consistent differences in the body size of MMP, connected to domestication, was detected.ConclusionsOur data demonstrate: 1- no evidence for the evolution of the size threshold for MMP in domesticated salmon, 2- the vastly lower incidence of MMP in domesticated strains under aquaculture conditions is primarily due to their genetically increased growth rate causing them to outgrow the size threshold for early maturation, 3- the incidence of MMP is likely to overlap among domesticated and wild salmon in the natural habitat where they typically display overlapping growth, although hybrid offspring may display lower incidences of mature male parr. These results have implications for wild salmon populations that are exposed to introgression from domesticated escapees.

How coexistence may influence life history: the reproductive strategies of sympatric congeneric terrestrial isopods (Crustacea, Oniscidea)

Patterns of allocation between reproduction, survival, and maintenance are what we call life history. By investigating the life-history strategy of sympatric species, we may understand how they are able to coexist, as different strategies are expected to evolve in species that occupy similar niche space. Terrestrial isopods are a group in which multiple species frequently inhabit the same area. Notably, they are usually infected by Wolbachia Hertig, 1936, a notorious manipulator of the hosts’ reproductive processes. Thus, the aim of this study was to analyze the investment in reproduction in three sympatric and closely related species of terrestrial isopods: Atlantoscia floridana (Van Name, 1940), Atlantoscia inflata Campos-Filho and Araujo, 2015, and Atlantoscia petronioi Campos-Filho, Contreira and Lopes-Leitzke, 2012, only the latter being infected with Wolbachia. We showed that the presence of the bacteria seems not to affect the fitness of A. petronioi, because there was no clear difference in the reproductive output of infected and noninfected individuals. On the other hand, we observed that the three species possess alternative life-history strategies; that is, they differ in how much they invest in maintenance (body size) and reproductive effort. Such differences probably facilitate the species coexistence, reducing the competition among them.

Conservation conundrum: Endangered predators eating endangered prey

Conservation conundrum: Endangered predators eating endangered prey

Alternative reproductive strategies in white-throated sparrows are associated with differences in parasite load following experimental infection.

Immune defences often trade off with other life-history components. Within species, optimal allocation to immunity may differ between the sexes or between alternative life-history strategies. White-throated sparrows (Zonotrichia albicollis) are unusual in having two discrete plumage morphs, white-striped and tan-striped. Within each sex, white-striped individuals are more aggressive and provide less parental care than tan-striped individuals. We extended immunocompetence handicap models, which predict sex differences in immunity and parasitism, to hypothesize that infection susceptibility should be greater in white-striped than tan-striped birds. We inoculated birds of both morphs with malarial parasites. Contrary to our prediction, among birds that became infected, parasite loads were higher in tan-striped than white-striped individuals and did not differ between the sexes. Circulating androgen levels did not differ between morphs but were higher in males than females. Our findings are not consistent with androgen-mediated immunosuppression. Instead, morph differences in immunity could reflect social interactions or life-history-related differences in risk of injury, and/or genetic factors. Although plumage and behavioural morphs of white-throated sparrow may differ in disease resistance, these differences do not parallel sex differences that have been reported in animals, and do not appear to be mediated by differences in androgen levels.