SEX ALLOCATION IN THE MONOECIOUS HERB BEGONIA SEMIOVATA

Sex-allocation models predict that the evolution of self-fertilization should result in a reduced allocation to male function and pollinator attraction in plants. The evolution of sex allocation may be constrained by both functional and genetic factors, however. We studied sex allocation and genetic variation for floral sex ratio and other reproductive traits in a Costa Rica population of the monoecious, highly selfing annual Begonia semiovata. Data on biomass of floral structures, flower sex ratios, and fruit set in the source population were used to calculate the average proportion of reproductive allocation invested in male function. Genetic variation and genetic correlations for floral sex ratio and for floral traits related to male and female function were estimated from the greenhouse-grown progeny of field-collected maternal families. The proportion of reproductive biomass invested in male function was low (0.34 at flowering, and 0.07 for total reproductive allocation). Significant among-family variation was detected in the size (mass) of individual male and female flowers, in the proportion of male flowers produced, and in the proportion of total flower mass invested in male flowers. Significant among-family variation was also found in flower number per inflorescence, petal length of male and female flowers, and petal number of female flowers. Except for female petal length, we found no difference in the mean value of these characters between selfed and outcrossed progeny, indicating that, with the possible exception of female petal length, the among-family variation detected was not the result of variation among families in the level of inbreeding. Significant positive phenotypic and broad-sense genetic correlations were detected between the mass of individual male and female flowers, between male and female petal length, and between number of male and number of female flowers per inflorescence. The ratio of stamen-to-pistil mass (0.33) was low compared to published data for autogamous species with hermaphroditic flowers, suggesting that highly efficient selfing mechanisms may evolve in monoecious species. Our results indicate that the study population harbors substantial genetic variation for reproductive characters. The positive genetic correlation between investment in male and female flowers may reflect selection for maximum pollination efficiency, because in this self-pollinating species, each female flower requires a neighboring male flower to provide pollen.     

Family affiliation,sex ratio and sporophyte frequency in unisexual mosses

Patterns of sex expression and sex ratios are key features of the life histories of organisms. Bryophytes are the only haploid‐dominant land plants. In contrast with seed plants, more than half of bryophyte species are dioecious, with rare sexual expression and sporophyte formation and a commonly female‐biased sex ratio. We asked whether variation in sex expression, sex ratio and sporophyte frequency in ten dioecious pleurocarpous wetland mosses of two different families was best explained by assuming that character states evolved: (1) in ancestors within the respective families or (2) at the species level as a response to recent habitat conditions. Lasso regression shrinkage identified relationships between family membership and sex ratio and sporophyte frequency, whereas environmental conditions were not correlated with any investigated reproductive trait. Sex ratio and sporophyte frequency were correlated with each other. Our results suggest that ancestry is more important than the current environment in explaining reproductive patterns at and above the species level in the studied wetland mosses, and that mechanisms controlling sex ratio and sporophyte frequency are phylogenetically conserved. Obviously, ancestry should be considered in the study of reproductive character state variation in plants. © 2013 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 174 , 163–172.     

Variation in male body size and reproductive allocation in the leafcutter ant <Emphasis Type="Italic">Atta colombica</Emphasis>: estimating variance components and possible trade-offs

Remarkably little is known about the traits that determine reproductive success of males in eusocial insects. Their window for mate choice decisions is very short, the actual mating process is very difficult to observe, and their small body sizes have likely prevented systematic studies in many species. In 2008 and 2009, we revisited a Panamanian population of Atta colombica leafcutter ants to partially repeat and complement a study of more than 15 years ago. We compared within- and between-colony variation in male body size (mass and width of head, mesosoma and gaster) and sperm characteristics (length, number and survival after exposure to saline buffer with and without added accessory gland secretion). We also measured the size of accessory glands as the main contributor of seminal fluid and the accessory testes containing all mature sperm, but we found few correlations between these variables. We also obtained little or no evidence for expected trade-offs between sperm number and sperm length and between mesosoma mass and sperm complement, although this could be due to limited sample size and unknown variation in larval resource allocation that was beyond our control. However, we found an interestingly bimodal distribution in broad-sense heritabilities (intra-class correlations) among the variables that we measured. Low heritabilities suggest that mesosoma size (mass and width), accessory testes size, sperm viability (measured as % survival in saline) and probably also accessory gland size are traits directly correlated with reproductive success. However, the much higher heritabilities for total body mass, gaster mass, head width, sperm length and sperm number suggest that these traits are less likely to make direct contributions to male fitness.     

Seasonal changes in circulating testosterone and androstenedione concentration and their correlation with the anomalous reproductive pattern in the male indian sheath-tailed bat, Taphozous longimanus

The relationship between reproductive organs, circulating testosterone, and androstenedione concentration in the male sheath-tailed bat, Taphozous longimanus, was studied. The masses of testis, accessory sex gland (prostate, ampullary), and epididymides showed three peaks, one each in October, January, and April. Monthly changes in testosterone also peaked during October, January, and April and closely coincided with the peak spermatogenesis. Serum androstenedione concentration peaked during November and January. Testosterone showed a strong correlation with masses of testis and accessory sex glands, while androstenedione showed strong correlation with the body mass. Different threshold levels of testosterone may be required to trigger spermatogenesis, secretory activity of accessory sex glands and mating in Taphozous longimanus and may be responsible for reproductive asynchrony in this species. Higher circulating concentrations of testosterone and androstenedione throughout the year in this species, as compared with other mammalian species, may be responsible for prolonged retention of sperm in the epididymides.     

Female reproductive patterns in nonhibernating bats

The major reproductive events in the oestrous cycles of nonhibernating mega- and microchiropteran species are reviewed. However, special attention is given to the reproductive biology of the temperate North American species, Tadarida brasiliensis mexicana (Molossidae), the Mexican free-tailed bat, which expresses dextral uterine and ovarian dominance. Only the larger right ovary is capable of producing an ovulatory follicle and the left has long been considered atrophic. In order to elucidate the normal oestrous cycle and define the structural and functional characteristics of the ovaries of this nonhibernating bat several analytical technique were utilized. These included light microscopic analysis, histochemical localization of delta 5-3 beta-hydroxysteroid dehydrogenase (HSD) activity, and radioimmunoassay of seasonal plasma progesterone levels. Interstitial tissue was found in both ovaries, but the left was almost entirely an interstitial organ. Histochemical analysis demonstrated that the gonads have seasonally varying amounts of delta 5-2 beta-HSD localized either in the thecal cells of the Graafian follicle or in the interstitium. The corpus luteum persisted throughout gestation, reaching maximum development just prior to parturition. Circulating progesterone levels correlated directly with luteal gland size and peaked at 106 ng/ml when the corpus luteum was largest.     

Resource allocation in a simultaneously hermaphroditic slug with phally polymorphism

The theory of resource allocation assumes that a resource not allocated to one function may be reallocated to another. Thus, in hermaphroditic species, an individual that suppresses the use of one sex function may free resources for the other sex function. We determined the relative importance of male copulatory organs in terms of their fraction of the total dry body weight and tested whether in the pulmonate land slug Deroceras laeve (Müller), individuals that lack the male copulatory organs (aphallics) reallocate this resource towards the female structures and/or towards life-history traits. To this end, we raised 13 families under uniparental reproduction and compared growth, length of the juvenile period, number of eggs produced, percentage of hatched eggs and hatching time among a- and euphallics. We also measured the reproductive and sex allocation of all individuals. Six out of 13 families contained no euphallic individuals. In the other seven families, the proportion of euphallic individuals ranged from 0.13 to 0.43. There was an enormous variation in life-history traits and reproductive and sex allocation among individuals, even among individuals of the same family. Allocation to the male function was very low in euphallic slugs (i.e. 1.35% of the total body dry mass and 12.33% of the total reproductive dry mass). Our results did not reveal a reallocation from the lost male function towards the female function, nor towards one of the life-history traits. Finally, we propose a scenario that could explain the maintenance of phally polymorphism in D. laeve.     

The hydraulic management of the Barra Bonita reservoir (SP, Brazil) as a factor influencing the temporal succession of its fish community.

The temporal succession of fish communities allows evaluating the environmental conditions and the adaptation capacity of the fish species to anthropogenic stress in reservoirs. The fish community at Barra Bonita reservoir was sampled in two different periods of the year (dry and rainy) and in three different areas of the reservoir (fluvial, transition, and lentic). The species list was compared to another four lists, trying to detect the transformations of the fish community for the last 15 years. In order to evaluate the adaptation of the present fish community to the hydraulic management of reservoir, the trophic and reproductive structures were studied. Temporal succession analysis shows little change in fish richness of the communities. The number of fish species varies between 23 and 39 for a total of 68 registered species. From this, 27 can be considered constant, 14 accessory and 27 accidental; the main differences observed were for Anostomidae, Loricariidae and Characidae families. In relation to the hydraulic management, we found a fish community stabilized and adapted to environmental stress. This is characterized by the dominance of small-sized fish species of opportunistic diet and high reproductive compensation (r-strategists). The overlap of biological cycles of the most abundant species with the reservoir level fluctuations points to the period from September to March-April as critical for reproductive success and only the species with partial reproductive strategy or parental care are best succeeded. These results, interpreted in the context of the reservoir aging process, indicate that Barra Bonita reservoir is entering a transition phase, between the colonization and aging stages.     

Sexual dimorphism, extrapair fertilizations, and operational sex ratio in great frigatebirds (Fregata minor)

Across taxa, the presence of sexual ornaments in one sex isusually correlated with disproportionately great parental effortby the other. Frigatebirds (Fregatidae) are sexually dimorphic,with males exhibiting morphological and behavioral ornaments,but males and females share in all aspects of parental effort.All other taxa in a clade of 237 species exhibit biparentalcare, but only frigatebirds exhibit pronounced sexual dimorphism. We tested for the presence of two factors that could contributeto the evolution of male ornaments in great frigatebirds: ahigh frequency of extrapair fertilizations and a male-biasedoperational sex ratio. In 92 families sampled over two breedingseasons, there was only one extrapair fertilization. However,in both seasons, there were more males than females availablefor mating, and the sex ratio among individuals actively engagedin mate-acquisition behavior was strongly male biased, withtypically five or six males available per female. Our resultssuggest that extrapair fertilizations are not responsible forthe exaggeration of sexual ornaments in male frigatebirds,and that operational sex ratio may be related to sexual dimorphismin this species. Further work is needed to determine whetherthe male-biased operational sex ratio creates the variancein male reproductive success that would be needed to drivethe evolution of male ornaments.     

Assessment of therapeutic responses to gametocytocidal drugs in Plasmodium falciparum malaria

Background

Effective mating between laboratory-reared males and wild females is paramount to the success of vector control strategies aiming to decrease disease transmission via the release of sterile or genetically modified male mosquitoes. However mosquito colonization and laboratory maintenance have the potential to negatively affect male genotypic and phenotypic quality through inbreeding and selection, which in turn can decrease male mating competitiveness in the field. To date, very little is known about the impact of those evolutionary forces on the reproductive biology of mosquito colonies and how they ultimately affect male reproductive fitness.

Methods

Here several male reproductive physiological traits likely to be affected by inbreeding and selection following colonization and laboratory rearing were examined. Sperm length, and accessory gland and testes size were compared in male progeny from field-collected females and laboratory strains of Anopheles gambiae sensu stricto colonized from one to over 25 years ago. These traits were also compared in the parental and sequentially derived, genetically modified strains produced using a two-phase genetic transformation system. Finally, genetic crosses were performed between strains in order to distinguish the effects of inbreeding and selection on reproductive traits.

Results

Sperm length was found to steadily decrease with the age of mosquito colonies but was recovered in refreshed strains and crosses between inbred strains therefore incriminating inbreeding costs. In contrast, testes size progressively increased with colony age, whilst accessory gland size quickly decreased in males from colonies of all ages. The lack of heterosis in response to crossing and strain refreshing in the latter two reproductive traits suggests selection for insectary conditions.

Conclusions

These results show that inbreeding and selection differentially affect reproductive traits in laboratory strains overtime and that heterotic ‘supermales’ could be used to rescue some male reproductive characteristics. Further experiments are needed to establish the exact relationship between sperm length, accessory gland and testes size, and male reproductive success in the laboratory and field settings.     

Variation of male reproductive apparatus in relation to fertilization modalities in the catfish families Auchenipteridae and Callichthyidae (Teleostei: Siluriformes)

To investigate the relationship between fertilization modalities and the morphology of male reproductive apparatus, two species of Auchenipteridae, Auchenipterus nuchalis and Tatia intermedia , and six species of Callichthyidae, Callichthys callichthys , Corydoras aeneus , Corydoras bondi , Corydoras ehrhardti , Corydoras potaroensis and Hoplosternum littorale were studied. The species analysed show either internal or external fertilization, the latter including the so called 'sperm drinking' type of mating. An anal fin modified as an intromittent organ, a pair of seminal vesicles, and the release of sperm in the form of discrete bundles (spermatozeugmata) characterize the male reproductive apparatus of the internal fertilizer A. nuchalis . Seminal vesicles are present also in C. aeneus , C. bondi , C. ehrhardti , C. potaroensis and H. littorale , species performing 'sperm drinking' spawning. In contrast, regardless of the family, species showing the more classic type of external fertilization lack specialized accessory organs. Where occurring, the major function of seminal vesicles is the secretion of mucins. The role of these mucosubstances, in relation to spermatozeugmata formation, in internal fertilizers, or the protection of sperm passing through the female gut, in the 'sperm-drinking' species, is discussed. Variation, between families, in the shape of testis and accessory structures, as well as in the type of spermatogenesis was found. Neither the reproductive modalities nor the phylogenetic relationships, however, appear to fully account for these differences.     

The evolution of sperm length in moths

Sperm form and function remain poorly understood despite being of fundamental biological importance. An instructive approach has been to examine evolutionary associations across comparable taxa between sperm characters and other, potentially selective reproductive traits. We adopt this approach here in a comparative study examining how sperm lengths are associated with male and female reproductive characters across moths. Primary data have revealed Lepidoptera to be an ideal order for examination: there is profound variation in the dimensions (but not organization) of the reproductive traits between closely related species which all share a monophyletic ancestry, for example, eupyrene sperm length varies from 110 to 12,675 microm. Eupyrene (normal fertilizing) and apyrene (anucleate and non-fertile) sperm lengths are positively correlated across taxa and both sperm types show positive associations with mating pattern (as measured by the residual testis size). At fertilization, eupyrene sperm must migrate down the often elongated female spermathecal duct from storage to unite with the ovum. Across taxa, the elongation of this duct is associated with increased eupyrene sperm length, suggesting a positive female influence on sperm size since longer, more powerful sperm may be selected to migrate and/or compete successfully down greater ductal lengths. Apyrene sperm length is not associated with female reproductive tract dimensions. However, we found a positive relationship between the residual testis volume and spermathecal volume, suggesting coevolution between male investment in spermatogenesis and the extent of the female sperm storage capacity. Within males, there is a positive association between the two organs which form the ejaculate-containing spermatophore: the testes and the accessory gland. The 'trade-up' in investment to these components is discussed in relation to paternal investment and mating patterns.     

Molecular dissection of nuptial gifts in divergent strains of Ostrinia moths

Seminal fluid proteins (SFPs) produced in the male accessory glands and ejaculatory duct are subject to strong sexual selection, often evolve rapidly and therefore may play a key role in reproductive isolation and species formation. However, little is known about reproductive proteins for species in which males transfer ejaculate to females using a spermatophore package. By combining RNA sequencing and proteomics, we characterize putative SFPs, identify proteins transferred in the male spermatophore and identify candidate genes contributing to a one‐way gametic incompatibility between Z and E strains of the European corn borer moth Ostrinia nubilalis. We find that the accessory glands and ejaculatory duct secrete over 200 highly expressed gene products, including peptidases, peptidase regulators and odourant‐binding proteins. A comparison between Ostrinia strains reveals that accessory gland and ejaculatory duct sequences with hormone degradation and peptidase activity are among the most extremely differentially expressed. However, most spermatophore peptides lack reproductive tissue bias or canonical secretory signal motifs and aproximately one‐quarter may be produced elsewhere before being sequestered by the male accessory glands during spermatophore production. In addition, most potential gene candidates for postmating reproductive isolation do not meet standard criteria for predicted SFPs and almost three‐quarters are novel, suggesting that both postmating sexual interactions and gametic isolation likely involve molecular products beyond traditionally recognized SFPs.     

Evolution of alternation of haploid and diploid phases in life cycles

Eukaryotic sex leads to an alternation of haploid and diploid nuclear phases. Because all multicellular animals are diploid, diploidy is often considered a 'biological success' and many arguments have been advanced to explain the evolution of a prolonged diploid phase. Nevertheless, among eukaryotes three basic situations are encountered, where the vegetative individuals are diploid or haploid or both. These three basic life cycles are widely distributed among kingdoms and in some taxa the occurrence of different life cycles within the same species has been reported. This article briefly summarizes the different hypotheses on the evolution of reproductive life cycles and underlines how possibilities of variation for this trait may open new perspectives for research.     

Reproductive cycles of male and female tuatara (Sphenodon punctatus) on Stephens Island, New Zealand

Cycles in gonadal activity and plasma sex steroid concentrations were investigated in wild female and male tuatara on Stephens Island, New Zealand. Females nest once every four years on average. Vitellogenesis is spread over the first three years, and mating, ovulation and nesting occur in the fourth. Oviducal eggs are carried for6–8 months before nesting. Although the length of this ovarian cycle is unparalleled among oviparous reptiles, the associated cycles in plasma concentrations of sex steroids are similar to those in other reptiles. Mean plasma concentrations of oestradiol increase during vitellogenesis, peak at mating, fall rapidly before or around ovulation, are low during most of gravidity, and rise slightly during late gravidity-nesting. Plasma concentrations of testosterone show a similarcycle. Mean plasma concentrations of progesterone are low during vitellogenesis and peak around ovulation. This periovulatory surge falls within1–2 months, and mean concentrations are low during the final 5–6 months of gravidity. Male tuatara show an annual, pre-nuptial reproductive cycle. Mean plasma concentrations of testosterone are low during winter, rise during spring, peak during midsummer-early autumn, when pre-nuptial displays and mating occur, and fall in mid autumn. Limited histological data indicate that spermiogenesis occurs during midsummer-early autumn, and also support a previous study suggesting that there may be no period of complete testicular and epididymal regression. In comparison with the pre-nuptial cycles of other temperate-zone reptiles, the cycle in male tuatara shows a more prolonged duration of testicular activity and of rising or elevated plasma testosterone concentrations. The prolonged reproductive cycles of male and female tuatara may be adaptations to a temperate environment with cool summers and cool, but not freezing, winters.     

Photoperiod influences the annual reproductive cycle of the male pallid bat (Antrozous pallidus)

Adult pallids bats collected in April or May, were maintained in short or long photoperiods (10 or 14 h light/day) for 3-6 months. In August, the short-day bats had regressed testes, epididymal spermatozoa and fully developed accessory sex glands, corresponding to the autumnal reproductive condition of field animals; long-day bats had testes undergoing spermatogenesis, few epididymal spermatozoa and undeveloped accessory sex glands (summer reproductive condition). Bats in each photoperiod manifested the expected autumnal reproductive pattern in October. We suggest that photoperiod influences the reproductive physiology of male pallid bats by affecting an endogenous circannual reproductive rhythm.     

Size structure, sex ratio and onset of sexual maturity of two species of Hypostomus

Length-frequency analyses showed that male Hypostomus affinis are bigger than females, but H. luetkeni showed no such differences. The sex ratio was estimated at 1:1 for both species. The temporal variation of sex ratio indicated a decrease in the male population when GSI was at its maximum value, suggesting that males are less prone to capture during the spawning season. According to the decrease of males during the reproductive period, paternal behaviour is proposed for both species. H. affinis reached sexual maturity at a smaller size than H. luetkeni. Considering the length weight relationship and the mortality rate of the reproductive females in both species, the physiological consequences of delay or precocity of the first maturation are discussed.     

Genetic and environmental sources of covariance among internal reproductive traits in the yellow dung fly

Substantial inter- and intraspecific variation is found in reproductive traits, but the evolutionary implications of this variation remain unclear. One hypothesis is that natural selection favours female reproductive morphology that allows females to control mating and fertilization and that diverse male reproductive traits arise as counter adaptations to subvert this control. Such co-evolution predicts the establishment of genetic correlations between male and female reproductive traits that closely interact during mating. Therefore, we measured phenotypic and genetic correlations between male and female reproductive tract characteristics in the yellow dung fly, Scathophaga stercoraria (Diptera: Scathophagidae), using a nested half-sib breeding experiment. We found significant heritabilities for the size of most reproductive tract traits investigated in both females (spermathecae and their ducts, accessory glands and their ducts) and males (testis size but not sperm length). Within the sexes, phenotypic and genetic correlations were mostly nil or positive, suggesting functional integration of or condition-dependent investment in internal reproductive traits. Negative intrasexual genetic correlations, potentially suggestive of resource allocation trade-offs, were not evident. Intersexual genetic correlations were mostly positive, reflecting expected allometries between male and female morphologies. Most interestingly, testis size correlated positively with female accessory gland size and duct length, potentially indicative of a co-evolutionary arms race. We discuss these and alternative explanations for these patterns of genetic covariance.     

Male reproductive allocation in fireflies (Photinus spp.)

Abstract. Resource allocation between somatic and reproductive structures has important fitness consequences for individuals, and optimal trade-offs are expected to depend not only on mating system differences among species but also on levels of resource stress within species. We tested the prediction that polyandry (associated with increased sperm competition) will increase male reproductive allocation in bioluminescent fireflies in Photinus spp. by comparing the relative mass of testes, seminal vesicles, and reproductive accessory glands among a monandrous and several polyandrous species. In addition, we examined a single population of a polyandrous species, Photinus greeni , to see how reproductive allocation might shift between years in response to different levels of larval resource stress. As predicted, males of P. collustrans , a monandrous species, showed a fivefold lower allocation to sperm production and a 100-fold lower allocation to reproductive accessory glands compared with males from polyandrous species. We also found evidence within P. greeni of a trade-off between allocation either to reproduction or to somatic tissue; following larval resource stress, males eclosed at significantly shorter body lengths, yet showed a 35% increase in their reproductive allocation. These results demonstrate that mating systems strongly influence male allocation to reproductive accessory glands as well as to sperm production. Furthermore, these results suggest that under larval resource stress males of Photinus spp. increase their allocation to reproduction at the expense of somatic tissue, thus maximizing their ability to produce nuptial gifts required for reproductive success.     

The function of female and male ornaments in the Inca Tern: evidence for links between ornament expression and both adult condition and reproductive performance

Inca Terns Larosterna inca are medium-size seabirds that breed along the Peruvian and Chilean coast. They are monogamous and both sexes incubate and contribute to chick provisioning. The sexes are similar in appearance and have elaborate ornaments, including a long white moustache of feathers and fleshy yellow wattles. In this paper we report the differences in ornamentation between sexes and examine whether the trait predicts body condition, reproductive performance or chick quality in either sex. The ornaments were similar in size and coloration between the sexes, except for the wattle length, the difference in which can be due to greater head length in males. Moustache length was the most reliable signal of body condition in both sexes. Moreover, there was a significant relationship between the moustache length and reproductive category of adults (non-breeder, unsuccessful breeder, or one or two chicks fledged). Both asymptotic chick body mass and the T-cell mediated response of chicks (a measure of immunocompetence) were related to the moustache length of male and female adults. These results provide support for the role of ornaments in mutual signalling of condition in this species. Female and male ornaments predict body condition, reproductive performance and chick quality, as predicted by sexual selection models.     

Life history variation in gametophyte populations of the moss Ceratodon purpureus (Ditrichaceae)

The life cycles of mosses and other bryophytes are unique among land plants in that the haploid gametophyte stage is free-living and the diploid sporophyte stage is ephemeral and completes its development attached to the maternal gametophyte. Despite predictions that populations of haploids might contain low levels of genetic variation, moss populations are characterized by substantial variation at isozyme loci. The extent to which this is indicative of ecologically important life history variation is, however, largely unknown. Gametophyte plants from two populations of the moss Ceratodon purpureus were grown from single-spore isolates in order to assess variation in growth rates, biomass accumulation, and reproductive output. The data were analyzed using a nested analysis of variance, with haploid sib families (gametophytes derived from the same sporophyte) nested within populations. High levels of life history variation were observed within both populations, and the populations differed significantly in both growth and reproductive characteristics. Overall gametophytic sex ratios did not depart significantly from 1:1 within either population, but there was significant variation among families in both populations for progeny sex ratio. Some families produced predominantly male gametophytes, while others yielded predominantly females. Because C. purpureus has a chromosomal mechanism of sex determination, these observations suggest differential (but unpredictable) germination of male and female spores. Life history observations showed that male and female gametophytes are dimorphic in size, maturation rates, and reproductive output.