Intense Male-male Competition Research Articles

Sexual color ornamentation, microhabitat choice, and thermal physiology in the common wall lizard (Podarcis muralis).

Common wall lizards (Podarcis muralis) in Italy show a striking variation in body coloration across the landscape, with highly exaggerated black and green colors in hot and dry climates and brown and white colors in cool and wet climates. Males are more intensely colored than females, and previous work has suggested that the maintenance of variation in coloration across the landscape reflects climatic effects on the strength of male-male competition, and through this sexual selection. However climatic effects on the intensity of male-male competition would need to be exceptionally strong to fully explain the geographic patterns of color variation. Thus, additional processes may contribute to the maintenance of color variation. Here we test the hypothesis that selection for green and black ornamentation in the context of male-male competition is opposed by selection against ornamentation because the genes involved in the regulation of coloration have pleiotropic effects on thermal physiology, such that ornamentation is selected against in cool climates. Field observations revealed no association between body coloration and microhabitat use or field active body temperatures. Consistent with these field data, lizards at the extreme ends of the phenotypic distribution for body coloration did not show any differences in critical minimum temperature, preferred body temperature, temperature-dependent metabolic rate, or evaporative water loss when tested in the laboratory. Combined, these results provide no evidence that genes that underlie sexual ornamentation are selected against in cool climate because of pleiotropic effects on thermal biology.

The origins and drivers of sexual size dimorphism in sharks.

While sexual size dimorphism (SSD) is abundant in nature, there is huge variation in both the intensity and direction of SSD. SSD results from a combination of sexual selection for large male size, fecundity selection for large female size and ecological selection for either. In most vertebrates, it is variation in the intensity of male-male competition that primarily underlies variation in SSD. In this study, we test four hypotheses regarding the adaptive value of SSD in sharks-considering the potential for each of fecundity, sexual, ecological selection and reproductive mode as the primary driver of variation in SSD between species. We also estimate past macroevolutionary shifts in SSD direction/intensity through shark phylogeny. We were unable to find evidence of significant SSD in early sharks and hypothesise that SSD is a derived state in this clade, that has evolved independently of SSD observed in other vertebrates. Moreover, there is no significant relationship between SSD and fecundity, testes mass or oceanic depth in sharks. However, there is evidence to support previous speculation that reproductive mode is an important determinant of interspecific variation in SSD in sharks. This is significant as in most vertebrates sexual selection is thought to be the primary driver of SSD trends, with evidence for the role of fecundity selection in other clades being inconsistent at best. While the phylogenetic distribution of SSD among sharks is superficially similar to that observed in other vertebrate clades, the relative importance of selective pressures underlying its evolution appears to differ.

Flanged males have higher reproductive success in a completely wild orangutan population.

Male orangutans (Pongo spp.) exhibit bimaturism, an alternative reproductive tactic, with flanged and unflanged males displaying two distinct morphological and behavioral phenotypes. Flanged males are larger than unflanged males and display secondary sexual characteristics which unflanged males lack. The evolutionary explanation for alternative reproductive tactics in orangutans remains unclear because orangutan paternity studies to date have been from sites with ex-captive orangutans, provisioning via feeding stations and veterinary care, or that lack data on the identity of mothers. Here we demonstrate, using the first long-term paternity data from a site free of these limitations, that alternative reproductive tactics in orangutans are condition-dependent, not frequency-dependent. We found higher reproductive success by flanged males than by unflanged males, a pattern consistent with other Bornean orangutan (Pongo pygmaeus) paternity studies. Previous paternity studies disagree on the degree of male reproductive skew, but we found low reproductive skew among flanged males. We compare our findings and previous paternity studies from both Bornean and Sumatran orangutans (Pongo abelii) to understand why these differences exist, examining the possible roles of species differences, ecology, and human intervention. Additionally, we use long-term behavioral data to demonstrate that while flanged males can displace unflanged males in association with females, flanged males are unable to keep other males from associating with a female, and thus they are unable to completely mate guard females. Our results demonstrate that alternative reproductive tactics in Bornean orangutans are condition-dependent, supporting the understanding that the flanged male morph is indicative of good condition. Despite intense male-male competition and direct sexual coercion by males, female mate choice is effective in determining reproductive outcomes in this population of wild orangutans.

How Female-Female Competition Affects Male-Male Competition: Insights into Postcopulatory Sexual Selection from Socially Polyandrous Species.

AbstractSexual selection is a major driver of trait variation, and the intensity of male competition for mating opportunities has been linked with sperm size across diverse taxa. Mating competition among females may also shape the evolution of sperm traits, but the effect of the interplay between female-female competition and male-male competition on sperm morphology is not well understood. We evaluated variation in sperm morphology in two species with socially polyandrous mating systems, in which females compete to mate with multiple males. Northern jacanas (Jacana spinosa) and wattled jacanas (J. jacana) vary in their degree of social polyandry and sexual dimorphism, suggesting species differences in the intensity of sexual selection. We compared mean and variance in sperm head, midpiece, and tail length between species and breeding stages because these measures have been associated with the intensity of sperm competition. We found that the species with greater polyandry, northern jacana, has sperm with longer midpieces and tails as well as marginally lower intraejaculate variation in tail length. Intraejaculate variation was also significantly lower in copulating males than in incubating males, suggesting flexibility in sperm production as males cycle between breeding stages. Our results indicate that stronger female-female competition for mating opportunities may also shape more intense male-male competition by selecting for longer and less variable sperm traits. These findings extend frameworks developed in socially monogamous species to reveal that sperm competition may be an important evolutionary force layered atop female-female competition for mates.

Effects of operational sex ratio and male density on size-dependent mating in Minshan’s toads, Bufo minshanicus, on the Tibetan Plateau of China

Abstract In many animal species, an increase in the operational sex ratio (OSR), density or a combination of both should lead to more intensive competition among individuals of the more abundant sex. To test this, we examined pairing patterns of Minshan’s toad (Bufo minshanicus) from six populations between 2008 and 2015 along the eastern Tibetan Plateau in south-west China. OSRs in breeding aggregations of Minshan’s toad are normally male biased and males actively compete with each other for acquisition and retention of mates. We found evidence that deviations from random mating by size varied between populations and between years according to the magnitude of the OSR and male density. Larger males were generally more successful in pairing than smaller males when the OSR was slightly male biased and male density was high. However, the resulting size-disproportionate mating was more evident when OSR was closer to 1.99, indicating a positive correlation with the intensity of aggressive scramble competition. Thus, the intensity of male-male competition may partly explain variation in size-disproportionate mating among populations.

The directional testes asymmetry increases with temperature in seven plateau brown frog (Rana kukunoris) populations

Environmental stress is generally regarded as an important evolutionary force for promoting the differentiation of shape, structure and function of animal organs closely related to survival and reproduction. Geographical variation of temperature and corresponding change in intensity of male-male competition might drive inter-population differences in directional testes asymmetry (DTA). Here, we investigated inter-population variation in DTA of the brown frog (Rana kukunoris) at seven different altitudes on the eastern Tibetan Plateau. We found that the size of right testes increased with temperature, but not left testes. We also found that male age, body mass or body condition, and testis mass had not effect on DTA, suggesting that heavier or older R. kukunoris males or those with larger testes had not stronger DTA. The operational sex ratio did not affect DTA, but there was a positive correlation between DTA and temperature, suggesting that differences in the length of activity period and resources availability across locations may affect the energy budget of this frog, resulting in a gradual change in reproduction energy parallel to increasing temperature.

Friend virus severity is associated with male mouse social status and environmental temperature

Pathogen virulence is highly variable within populations, and although many factors contributing to virulence differences are known, there is still much variation left unexplained. Identifying and characterizing environmental conditions associated with different virulence levels is therefore an important undertaking in infectious disease research. One factor considered to be a major determinant of overall health and susceptibility to disease in social animals is social status. Health differences associated with social status are thought to be caused by different levels of chronic stress in higher- versus lower-status individuals. There is considerable evidence that these effects extend to the standing immune profile and that social status directly influences susceptibility to pathogens. Here we examined the association between dominance status in male wild-derived house mice, Mus musculus, and susceptibility to Friend virus complex in the context of seminatural populations with intense male-male competition and no predation. Due to an interruption in our facility's heating system, we were unexpectedly presented with the opportunity to assess how reduced ambient temperature influences the association of host social status and pathogen virulence. Environmental temperature has been implicated as a contributor to pathogen virulence, giving us a unique chance to examine its role in a previously unexamined pathogen system, while the added context of social status can expand our understanding of how the interaction of different environmental conditions affects virulence. We found that pathogen virulence and replication were lower in socially dominant hosts compared to nondominant hosts. When temperature was reduced, cool enclosure-housed dominant males were more susceptible to infection than their warm enclosure-housed counterparts. The mechanistic underpinnings that link infectious disease and social status remain difficult to disentangle from their associated factors, but this study opens the door for future experiments using a novel approach in the most well-studied mammalian model available.

Life history, mating dynamics and the origin of parental care.

Parental care, mating dynamics and life history co-evolve. Understanding the diversity of reproductive patterns found in nature is a major focus of evolutionary ecology research. Previous research suggests that the origin of parental care of eggs will be favoured when egg and adult death rates and juvenile survival are relatively high. However, the previous research that explored the link between care and life history did not account for among-species variation in mating dynamics. As mating dynamics are generally expected to influence care, we explore, theoretically, the life-history conditions (stage-specific rates of maturation and survival) that favour parental care across three mating scenarios: reproductive rate (1) is unaffected by males (assuming that some males are present), (2) increases as male abundance increases or (3) decreases as male abundance increases. Across scenarios, all forms of care were most strongly favoured when egg and adult death rates, juvenile survival and female egg maturation rates were relatively high. When reproductive rate was unaffected by male abundance or increased as male abundance increased, as we might expect in systems in which females are mate-limited, all forms of care were most strongly favoured when male egg maturation rate (i.e. the rate at which male eggs develop, mature and hatch) was moderate or high. When greater male abundance inhibited reproduction, which might occur in systems with intense male-male competition, all forms of care were most strongly favoured when male egg maturation rate was low-to-moderate. These results suggest that life history affects the evolution of parental care, and sex-specific life history can interact with mating dynamics to influence the origin of care.

Oral processing, sexual selection, and size variation in the circumorbital region of Colobus and Piliocolobus.

The function of the browridge in primates is a subject of enduring debate. Early studies argued for a role in resisting masticatory stresses, but recent studies have suggested sexual signaling as a biological role. We tested associations between circumorbital form, diet, oral processing, and social behavior in two species of colobus monkey-the king colobus (Colobus polykomos) and western red or bay colobus (Piliocolobus badius). We quantified circumorbital size and dimorphism in a sample of 98 crania. Controlling for age and facial size, we tested whether variation in circumorbital morphology can be explained by variation in diet, oral processing behavior, masticatory muscle size, and mating system. To contextualize our results, we included a broader sample of facial dimorphism for 67 anthropoid species. Greater circumorbital thickness is unrelated to the stresses of food processing. King colobus engages in longer bouts of anterior tooth use, chews more per ingestive event, and processes a tougher diet, yet circumorbital thickness of C. polykomos is reduced compared to P. badius. Differences in circumorbital development do not vary with wear or facial size. Greater sexual dimorphism is present in P. badius; comparisons across anthropoids indicated patterns of circumorbital dimorphism were decoupled from overall size dimorphism. The expanded circumorbits of male red colobus monkeys evolved in response to intense male-male competition. This hypothesis is consistent with the pattern across anthropoid primates and highlights the underappreciated role of sexual selection in shaping the primate face.

Seasonality and interindividual variation in mandrill feeding ecology revealed by stable isotope analyses of hair and blood.

Mandrills are large-bodiedterrestrial forest primates living in particularly large social groups of several hundred individuals. Following these groups in the wild to assess differences in diet over time as well as among individuals is demanding. We here use isotope analyses in blood and hair obtained during repeated captures of 43 identified free-ranging mandrills (Mandrillus sphinx) from Southern Gabon, to test how dietary variation relates to the season as well as an individual's age and sex. We measured the stable carbon (δ13 C‰) and nitrogen (δ15 N‰) isotope ratios in 46 blood and 214 hair section samples as well as from a small selection of mandrill foods (n = 24). We found some seasonal isotopic effects, with lower δ13 C values but higher δ15 N values observed during the highly competitive long dry season compared to the fruit-rich long rainy season. Variation in δ13 C was further predicted by individual age, with higher δ13 C values generally found in younger individuals suggesting that they may consume more high canopy fruit than older individuals, or that older individuals consume more low canopy foliage. The best predictor for δ15 N values was the interaction between age and sex, with mature and reproductively active males revealing the highest δ15 N values, despite the observation that males consume substantially less animal food items than females. We interpret high δ15 N values in these mature male mandrill blood and hair sections to be the result of nutritional stress associated with intense male-male competition, particularly during mating season. This is the first study showing isotopic evidence for nutritional stress in a free-ranging primate species and may spark further investigations into male mandrill diet and energy balance.

Immune challenge of mating effort: steroid hormone profile, dark ventral patch and parasite burden in relation to intrasexual competition in male Iberian red deer.

Testosterone secretion may regulate the reproductive effort and the development of sexual traits, but it may also involve costs at the immunological and metabolic levels. However, the evidence for this trade-off in wild populations is scarce. Cortisol also plays an important role in mediating the reproductive and immune functions. In this study, we analyzed whether the endoparasite burden relates to hormonal levels (fecal testosterone and cortisol metabolites) and/or morphological sexual traits (size of the dark ventral patch, a trait that indicates reproductive effort in males) in male Iberian red deer. For this purpose, we sampled male red deer harvested during hunting actions in 2 types of populations in south western Spain that differed in structure, affecting the level of male-male competition for mates. We used coprological analyses to estimate the parasite burden mainly of gastrointestinal and bronchopulmonary nematodes and of protozoa, and assessed testosterone and cortisol metabolite levels from fecal pellets. We found a positive relationship of host parasitation with both testosterone levels and the size of the dark ventral patch, but these relationships depended on the intensity of male-male competition in the population, being only found under the high-competition scenario. These results are discussed under the hypothesis of the testosterone immunocompetence handicap, suggesting a cost at the immunological level, and, therefore, higher susceptibility to parasite infection in males that make a greater reproductive effort. However, this effect seems to be modulated by the social environment (male-male competition) that might lead to different optima in testosterone production and sexual trait development.

Shifts in Male Reproductive Tactics over the Life Course in a Polygynandrous Mammal.

In polygynous and polygynandrous species, there is often intense male-male competition over access to females, high male reproductive skew, and more male investment in mating effort than parenting effort [1]. However, the benefits derived from mating effort and parenting effort may change over the course of males' lives. In many mammalian species, there is a ∩-shaped relationship between age, condition, and resource holding power as middle-aged males that are in prime physical condition outcompete older males [2-8] and sire more infants [9-12]. Thus, males might derive more benefits from parenting effort than mating effort as they age and their competitive abilities decline [13]. Alternatively, older males may invest more effort in making themselves attractive to females as mates [14]. One way that older males might do so isby developing relationships with females and providing care for their offspring [14, 15]. Savannah baboons provide an excellent opportunity to test these hypotheses. They form stable multi-male, multi-female groups, and males compete for high ranking positions. In yellow and chacma baboons (Papio cynocephalus and P.ursinus), there is a ∩-shaped relationship between male age and dominance rank [12], and high rank enhances paternity success [12,16]. Lactating female baboons form close ties ("primary associations" hereafter) with particular males [15-20], who support them and their infants in conflicts [15, 19] and buffer their infants from rough handling [20]. Females' primary associates are often, but not always, the sires of their current infants [16,20-22].

Male-male competition and repeated evolution of terrestrial breeding in Atlantic Coastal Forest frogs.

Terrestrial breeding is a derived condition in frogs, with multiple transitions from an aquatic ancestor. Shifts in reproductive mode often involve changes in habitat use, and these are typically associated with diversification in body plans, with repeated transitions imposing similar selective pressures. We examine the diversification of reproductive modes, male and female body sizes, and sexual size dimorphism (SSD) in the Neotropical frog genera Cycloramphus and Zachaenus, both endemic to the Atlantic rainforest of Brazil. Species in this clade either breed in rocky streams (saxicolous) or in terrestrial environments, allowing us to investigate reproductive habitat shifts. We constructed a multilocus molecular phylogeny and inferred evolutionary histories of reproductive habitats, body sizes, and SSD. The common ancestor was small, saxicolous, and had low SSD. Terrestrial breeding evolved independently three times and we found a significant association between reproductive habitat and SSD, with shifts to terrestrial breeding evolving in correlation with decreases in male body size, but not female body size. Terrestrial breeding increases the availability of breeding sites and results in concealment of amplexus, egg-laying, and parental care, therefore reducing male-male competition at all stages of reproduction. We conclude that correlated evolution of terrestrial reproduction and small males is due to release from intense male-male competition that is typical of exposed saxicolous breeding.

Plant-pollinator interactions along the pathway to paternity.

The male fitness pathway, from pollen production to ovule fertilization, is thought to strongly influence reproductive trait evolution in animal-pollinated plants. This pathway is characterized by multiple avenues of pollen loss which may lead to reductions in male fitness. However, empirical data on the mechanistic processes leading to pollen loss during transport are limited, and we therefore lack a comprehensive understanding of how male fitness is influenced by each step in the pollination process. This review assesses the history of studying male function in plants and identifies critical gaps in our understanding of the ecology and evolution of pollen transport. We explore male reproductive function along the steps of the pathway to paternity and discuss evolutionary options to overcome barriers to siring success. In particular, we present a newly emerging idea that bodies of pollinators function as a dynamic arena facilitating intense male-male competition, where pollen of rival males is constantly covered or displaced by competitors. This perspective extends the pollen-competitive arena beyond the confines of the stigma and style, and highlights the opportunity for important new breakthroughs in the study of male reproductive strategies and floral evolution.

The role of copulatory plugs in the mosquito-parasitic nematode Strelkovimermis spiculatus

The mosquito-parasitic nematode, Strelkovimermis spiculatus (Mermithidae: Nematoda) emerges from hosts and aggregates to form mating clusters characterized by intense male-male competition for females. Successful males deposit an adhesive copulatory plug over the female vulva after insemination. In choice experiments, males strongly preferred virgin females, whereas plugged females were ignored. Males made no attempt to displace copulatory plugs deposited by previous males. Plugged females repelled males without the need for physical contact. The observed chemical repellency was independent of females, since excised plugs alone showed the same negative male response. Removal of the plug shortly after mating reduced fecundity by 90%, presumably due to spermatids leaking from the vulva. The plug as a nutritional gift hypothesis was rejected because there was no post-mating reduction in plug size that would have indicated absorption. Despite being a male adaption to sperm competition, we conclude that the copulatory plug serves the fitness needs of both males and females in multiple roles that include paternity preservation, reduced male harassment, improved male search for mates, increased fecundity, and sperm retention.

Contexts and consequences of takeovers in hamadryas baboons: Female parity, reproductive state, and observational evidence of pregnancy loss.

The evolutionary consequences of sexual selection and sexual conflict are epitomized in the hamadryas baboon, a species characterized by strong sexual dimorphism and intense male-male competition. Hamadryas males coerce individual females into reproductively exclusive one-male units via aggressive takeovers, and infants involved in such takeovers are at least four times more likely to be killed (or otherwise die) after takeovers compared to other times. Here we examine female reproductive state before and after takeovers to further investigate the determinants of takeovers and their impact on female reproduction. Our data set consists of 162 takeovers over a period of 20 years in a population of wild hamadryas baboons at Filoha, Ethiopia. Results show that, in contrast to their polygynandrous relatives, hamadryas males show no bias with regard to female reproductive state at the time of the takeover, but are more likely to take over nulliparous compared to multiparous females. In addition, these results reinforce previous findings demonstrating the high likelihood of infant loss after hamadryas takeovers and suggest that females may use an additional strategy of pregnancy termination to curtail investment in offspring that are very likely to die anyway. Thus, hamadryas males may target females with high future reproductive value and hamadryas females may employ counterstrategies to help mitigate the effects of sexual coercion and infanticide on their own fitness.

On the relative effect of spawning asynchrony, sperm quantity, and sperm quality on paternity under sperm competition in an external fertilizer

How much of a fitness benefit is obtained by dominant males of external fertilizers from releasing ejaculates in synchrony with female egg-release when engaging in sperm competition, and what is the most important sperm trait for paternity in these situations? The Arctic charr (Salvelinus alpinus) is an external fertilizer experiencing intense male-male competition over reproductive opportunities including sperm competition. To compensate for their disadvantage the sneaker males, which often spawn out of synchrony with the female, produce more and faster sperm than the guarding males. We used controlled in vitro fertilization trials with experimentally produced dominant and subordinate, sneaker males to test what effect relative synchrony in gamete release, sperm quality (i.e., motility and velocity) and sperm quantity have on a male’s fertilization success in pair-wise sperm competitions. When the sneaker males released ejaculates after the guarding male there was no overall difference in fertilization success. The quality (i.e., motility and velocity) of a male’s sperm relative to that of the competing male was the best predictor of male fertilization success regardless of their mating tactic and spawning synchrony. The relative number of sperm cells also had an effect on fertilization success, but mainly when the dominant and sneaker male ejaculated synchronously. Our close imitation of natural sperm competition in charr shows that the sneaker males of external fertilizing species may fully compensate for their disadvantaged mating role by producing ejaculates of higher quality - an adjustment strangely not met by dominants.

Dimorfismo sexual en tamaño y marca frontal en el chorlo nevado (Charadrius nivosus)

El dimorfismo sexual en aves se expresa de diferentes formas, entre ellas está el tamaño o la coloración del plumaje. En aves playeras las marcas melánicas son distintas entre sexos y muy variables; se cree que cumplen una función disruptiva con el paisaje. Analizamos el dimorfismo sexual en tamaño corporal y en la estructura de la barra frontal del chorlo nevado (Charadrius nivosus) y encontramos que el tarso, la cuerda alar y que tanto la longitud como la amplitud de la barra frontal son de mayores dimensiones en los machos. Además, describimos por primera vez un ornamento con forma de “cuernos” en la frente, exclusivo de los machos. Aunque otros autores han revisado la variabilidad de estas marcas, su valor adaptativo es poco conocido. Consideramos que el dimorfismo en la estructura de la barra frontal es importante dentro y entre sexos durante la temporada reproductora, ya que cumple un papel clave durante los despliegues de cortejo y en la competencia entre machos, donde son exhibidas las plumas del pecho que contrastan con esta marca. Además, las medidas morfométricas mayores en los machos son consistentes con la poliandria y la competitividad intensa entre los machos.

Using visual modelling to study the evolution of lizard coloration: sexual selection drives the evolution of sexual dichromatism in lacertids

Sexual selection has been invoked as a major force in the evolution of secondary sexual traits, including sexually dimorphic colourations. For example, previous studies have shown that display complexity and elaborate ornamentation in lizards are associated with variables that reflect the intensity of intrasexual selection. However, these studies have relied on techniques of colour analysis based on human--rather than lizard--visual perception. Here, we use reflectance spectrophotometry and visual modelling to quantify sexual dichromatism considering the overall colour patterns of lacertids, a lizard clade in which visual signalling has traditionally been underrated. These objective methods of colour analysis reveal a large, previously unreported, degree of sexual dichromatism in lacertids. Using a comparative phylogenetic approach, we further demonstrate that sexual dichromatism is positively associated with body size dimorphism (an index of intrasexual selection), suggesting that conspicuous coloration in male lacertids has evolved to improve opponent assessment under conditions of intense male-male competition. Our findings provide the first evidence for the covariation of sexual dichromatism and sexual size dimorphism in lacertids and suggest that the prevalent role of intrasexual selection in the evolution of ornamental coloration is not restricted to the iguanian lineage, but rather may be a general trend common to many diurnal lizards.

Alternate Reproductive Tactics in an African Dung Beetle, Circellium bacchus (Scarabeidae)

Male-male competition is recognized as a potent force of sexual selection. When intra-sexual competition is strong then selection theory predicts that alternative male phenotypes will evolve. Circellium bacchus is a large, hornless, ball-rolling dung beetle with extensive variation in size and subject to intense male-male contest competition. We proposed that small male C. bacchus, which are unlikely to be successful in male contest competition, would adopt the alternative reproductive tactic of sneaking copulations. This alternative tactic is likely to influence not only behavior, but morphology with an expectation that sneaking males would invest more resources in testes development. Investigation of testis allometry revealed that smaller male C. bacchus beetles had relatively larger testes than their bigger conspecifics. Furthermore, as resources may be limited during larval development, differential investment in testes development should result in adult male beetles either competing for fertilizations or for access to mates. This is seen in C. bacchus as two alternate male phenotypes; smaller beetles with a relatively low body mass invest proportionately more in testes development compared to larger, heavier form in which testes size does not scale with condition. To our knowledge this study provides the first investigation of alternative phenotypes in the reproductive tactics of a ball-rolling dung beetle species.