Studies Of Life-history Evolution Research Articles

Syngnathoid Evolutionary History and the Conundrum of Fossil Misplacement.

Seahorses, pipefishes, trumpetfishes, shrimpfishes, and allies are a speciose, globally distributed clade of fishes that have evolved a large number of unusual body plans. The clade that includes all these forms, Syngnathoidei, has become a model for the study of life history evolution, population biology, and biogeography. Yet, the timeline of syngnathoid evolution has remained highly contentious. This debate is largely attributable to the nature of the syngnathoid fossil record, which is both poorly described and patchy for several major lineages. Although fossil syngnathoids have been used to calibrate molecular phylogenies, the interrelationships of extinct species and their affinities to major living syngnathoid clades have scarcely been quantitatively tested. Here, I use an expanded morphological dataset to reconstruct the evolutionary relationships and clade ages of fossil and extant syngnathoids. Phylogenies generated using different analytical methodologies are largely congruent with molecular phylogenetic trees of Syngnathoidei but consistently find novel placements for several key taxa used as fossil calibrators in phylogenomic studies. Tip-dating of the syngnathoid phylogeny finds a timeline for their evolution that differs slightly from the one inferred using molecular trees but is generally congruent with a post-Cretaceous diversification event. These results emphasize the importance of quantitatively testing the relationships of fossil species, particularly when they are critical to assessing divergence times.

Density-dependent effects of mortality on the optimal body size to shift habitat: Why smaller is better despite increased mortality risk.

Many animal species across different taxa change their habitat during their development. An ontogenetic habitat shift enables the development of early vulnerable‐to‐predation stages in a safe “nursery” habitat with reduced predation mortality, whereas less vulnerable stages can exploit a more risky, rich feeding habitat. Therefore, the timing of the habitat shift is crucial for individual fitness. We investigate the effect that size selectivity in mortality in the rich feeding habitat has on the optimal body size at which to shift between habitats using a population model that incorporates density dependence. We show that when mortality risk is more size dependent, it is optimal to switch to the risky habitat at a smaller rather than larger body size, despite that individuals can avoid mortality by staying longer in the nursery habitat and growing to safety in size. When size selectivity in mortality is high, large reproducing individuals are abundant and produce numerous offspring that strongly compete in the nursery habitat. A smaller body size at habitat shift is therefore favored because strong competition reduces growth potential. Our results reveal the interdependence among population structure, density dependence, and life history traits, and highlight the need for integrating ecological feedbacks in the study of life history evolution.

Latitudinal Variation in Life History Reveals a Reproductive Advantage in the Texas Horned Lizard (Phrynosoma cornutum)

Geographically widespread species that occupy varied thermal environments provide testable models for understanding the evolution of life-history responses to latitude. Studies that draw range-wide conclusions using descriptive data from populations in the core of a species' distribution can overlook meaningful inter-population variation. The Texas Horned Lizard (Phrynosomatidae: Phrynosoma cornutum) spans an extensive latitudinal distribution in North America and has been well studied in connection with life-history evolution, yet populations occupying the most northern and coldest areas within its range were absent from previous examinations. We tested genus-wide patterns and challenged species-specific findings on the evolution of the life-history strategy of P. cornutum using populations at the northern edge of its geographic range and comparative material from farther south. Traits in populations at the highest latitudes corroborated several patterns for the genus and species, including delayed reproduction and a trend towards smaller adult body size with increasing latitude. Novel in our study, however, was the finding of a comparative increase in clutch size among size-adjusted females in Kansas, indicating a reproductive response for greater fecundity at the northern edge of its geographic range. Furthermore, analyses adjusted for body size revealed that egg dimensions were constant across variation in clutch size, suggesting that there is not a strong relationship between egg size and egg number across latitude. We discuss the selective pressures that may have resulted in the diminution of adult body size coupled with greater fecundity that is unique to the northernmost populations of P. cornutum. Our findings highlight the type of insights into the study of life-history evolution that can be gained across Phrynosomatidae from the inclusion of populations representing latitudinal endpoints.

Population-Level Life History in Italy, Spain and Mexico: The Impact of Regional Climate, Parasite Burden, and Population Density

Theoretical and empirical work in the study of life history evolution has largely shifted away from density-dependent models since the 1980s, focusing on alternative causative processes such as the prominence of extrinsic morbidity and mortality. Nevertheless, recent evidence suggests that density dependent models are integral, even if clearly not sufficient, to account for trajectories of life history evolution. However, little investigation has been conducted among human populations to compare these models. Here we collate data for subnational regions of Italy, Spain, and Mexico to directly compare alternative models of life history evolution, including information on several dimensions of climate, parasite burden, population density, and manifest indicators of life history strategy. We examine direct and indirect effects of potential predictors upon life history speed, and address the hierarchical relations among these variables in relation to the goal of theory integration. Results revealed that ecological indicators had no significant impact on population density. Population density and the brumal factor (a composite denoting higher altitude, latitude and colder mean temperatures) predicted slower life histories, however parasite burden (a measure of morbidity and mortality) failed to exhibit a direct effect upon life history speed. The impact of ecological factors was stronger for Italy and Spain, in comparison to Mexico. We discuss the implications for life history theory.

Synchronous and Non-Synchronous Semelparity in Sibling Species of Pulmonates.

Diverse life histories have been documented in terrestrial pulmonates, which inhabit different regions in climate. Life history traits are often phenotypically plastic and vary depending on the environment. Thus, surveys using designs that control for the confounding effects of environment are needed to evaluate the evolutionary differences between populations of closely related species in the wild. We examined the life histories of sibling species of terrestrial pulmonate within two regions of similar climates. Bradybaena pellucida (BP) is endemic to Japanese islands, and has recently been expanding its distribution northeastward, whereas B. similaris (BS) has been introduced by humans into temperate and tropical regions worldwide. We found that these species exhibit discrete differences in population dynamics and life cycle, despite their close relatedness. The annual life cycle of BP is synchronized among individuals in a population. Thus, BP is univoltine with discontinuous generation. In contrast, BS individuals do not synchronize their growth or reproduction, and thus exhibit overlapping generations. Our results indicate that synchronized and non-synchronized population dynamics diverge relatively rapidly in semelparous pulmonates. This type of difference has not been documented in pulmonate life history, and may have been overlooked because only a few studies have explicitly compared life cycles of closely related species within the same climate. Our results provide a basis for further studies of life history evolution in pulmonates.

The plastic fly: the effect of sustained fluctuations in adult food supply on life-history traits.

Many adult traits in Drosophila melanogaster show phenotypic plasticity, and the effects of diet on traits such as lifespan and reproduction are well explored. Although plasticity in response to food is still present in older flies, it is unknown how sustained environmental variation affects life-history traits. Here, we explore how such life-long fluctuations of food supply affect weight and survival in groups of flies and affect weight, survival and reproduction in individual flies. In both experiments, we kept adults on constant high or low food and compared these to flies that experienced fluctuations of food either once or twice a week. For these ‘yoyo’ groups, the initial food level and the duration of the dietary variation differed during adulthood, creating four ‘yoyo’ fly groups. In groups of flies, survival and weight were affected by adult food. However, for individuals, survival and reproduction, but not weight, were affected by adult food, indicating that single and group housing of female flies affects life-history trajectories. Remarkably, both the manner and extent to which life-history traits varied in relation to food depended on whether flies initially experienced high or low food after eclosion. We therefore conclude that the expression of life-history traits in adult life is affected not only by adult plasticity, but also by early adult life experiences. This is an important but often overlooked factor in studies of life-history evolution and may explain variation in life-history experiments.

Replicated high-density genetic maps of two great tit populations reveal fine-scale genomic departures from sex-equal recombination rates.

Linking variation in quantitative traits to variation in the genome is an important, but challenging task in the study of life-history evolution. Linkage maps provide a valuable tool for the unravelling of such trait−gene associations. Moreover, they give insight into recombination landscapes and between-species karyotype evolution. Here we used genotype data, generated from a 10k single-nucleotide polymorphism (SNP) chip, of over 2000 individuals to produce high-density linkage maps of the great tit (Parus major), a passerine bird that serves as a model species for ecological and evolutionary questions. We created independent maps from two distinct populations: a captive F2-cross from The Netherlands (NL) and a wild population from the United Kingdom (UK). The two maps contained 6554 SNPs in 32 linkage groups, spanning 2010 cM and 1917 cM for the NL and UK populations, respectively, and were similar in size and marker order. Subtle levels of heterochiasmy within and between chromosomes were remarkably consistent between the populations, suggesting that the local departures from sex-equal recombination rates have evolved. This key and surprising result would have been impossible to detect if only one population was mapped. A comparison with zebra finch Taeniopygia guttata, chicken Gallus gallus and the green anole lizard Anolis carolinensis genomes provided further insight into the evolution of avian karyotypes.

Intra-Population Genetic Variation in the Temporal Pattern of Egg Maturation in a Parasitoid Wasp

Parasitoid wasps are taxonomically and biologically extremely diverse. A conceptual framework has recently been developed for understanding life-history evolution and diversification in these animals, and it has confirmed that each of two linked life-history traits – the mode of larval development and the temporal pattern of egg maturation – acts as an organiser of life-history. The framework has been predicated on the assumption that there exists sufficient genetic variation in the latter trait to allow it to be shaped by natural selection. Focusing on the parasitoid wasp Trichogramma brassicae, our aim was to test the validity of that assumption, using established quantitative genetic methods. We demonstrate the existence of a statistically significant degree of intra-population polygenic variation in the temporal pattern of egg production within the wasp population we studied. Furthermore, our results, together with published data on clinal variation in the egg maturation pattern of another species, suggest that intra-specific evolutionary shifts in the temporal pattern of egg maturation of parasitoid wasps can result from a change in allocation to egg production either before, or very shortly after adult emergence, without there being an accompanying change in lifetime fecundity. As well as opening new avenues of research into the reproductive strategies, behaviour, community organisation and biological control potential of parasitoid wasps, this discovery also has implications for studies of life-history evolution and diversification in insects generally.

Larval life history and anti-predator strategies are affected by breeding phenology in an amphibian

Seasonal time constraints can pose strong selection on life histories. Time-constrained animals should prioritise fast development over predation risk to avoid unfavourable growing conditions. However, changes in phenology could alter the balance between anti-predator and developmental needs. We studied variation of anti-predator strategies in common frog (Rana temporaria) tadpoles in four populations from the two extremes of a latitudinal gradient across Sweden. We examined, under common conditions in the laboratory, the anti-predator responses and life histories of tadpoles raised with predatory Aeshna dragonfly larvae in two consecutive years with a difference of 20 days in breeding time in the north, but no difference in breeding time in the nouth. In a year with late breeding, northern tadpoles did not modify their behaviour and morphology in the presence of predators, and metamorphosed faster and smaller than tadpoles born in a year with early breeding. In the year with early breeding, northern tadpoles showed a completely different anti-predator strategy by reducing activity and developing morphological defences in the presence of predators. We discuss the possible mechanisms that could activate these responses (likely a form of environmentally-mediated parental effect). To our knowledge, this is the first study to show that a vertebrate modifies the anti-predator strategy of its offspring in response to natural variation in reproductive phenology, which highlights the need to consider phenology in studies of life-history evolution.

Transcriptome Analysis and SNP Development Can Resolve Population Differentiation of Streblospio benedicti, a Developmentally Dimorphic Marine Annelid

Next-generation sequencing technology is now frequently being used to develop genomic tools for non-model organisms, which are generally important for advancing studies of evolutionary ecology. One such species, the marine annelid Streblospio benedicti, is an ideal system to study the evolutionary consequences of larval life history mode because the species displays a rare offspring dimorphism termed poecilogony, where females can produce either many small offspring or a few large ones. To further develop S. benedicti as a model system for studies of life history evolution, we apply 454 sequencing to characterize the transcriptome for embryos, larvae, and juveniles of this species, for which no genomic resources are currently available. Here we performed a de novo alignment of 336,715 reads generated by a quarter GS-FLX (Roche 454) run, which produced 7,222 contigs. We developed a novel approach for evaluating the site frequency spectrum across the transcriptome to identify potential signatures of selection. We also developed 84 novel single nucleotide polymorphism (SNP) markers for this species that are used to distinguish coastal populations of S. benedicti. We validated the SNPs by genotyping individuals of different developmental modes using the BeadXPress Golden Gate assay (Illumina). This allowed us to evaluate markers that may be associated with life-history mode.

The contribution of developmental palaeontology to extensions of evolutionary theory

AbstractEvo‐devo is featuring prominently in current discussion to extend evolutionary theory. Developmental palaeontology, the study of life history evolution and ontogeny in fossils, remains an area of investigation that could benefit from, but also illuminate, the discourse and research agenda of evo‐devo. Understanding how and why evolution proceeds in phenotypic space is an important goal of evo‐devo and one that can be significantly enriched through the examination of development in the fossil record (Palaeo‐evo‐devo). Such an approach permits developmental pathways to be extended into the past, constraining hypotheses of developmental evolution in ways that cannot be predicted by patterns observed from extant taxa alone. The comparison of developmental dynamics among extant and extinct taxa yields a more complete understanding of the temporal persistence of factors that shape evolution in phenotypic space. As more data are compiled that document ‘fossilized ontogenies’, a stage will emerge from which insights into the evolution of development can begin to appraise those phenotypes that are inaccessible to evo‐devo.

Short‐ and long‐term consequences of early developmental conditions: a case study on wild and domesticated zebra finches

Divergent selection pressures among populations can result not only in significant differentiation in morphology, physiology and behaviour, but also in how these traits are related to each other, thereby driving the processes of local adaptation and speciation. In the Australian zebra finch, we investigated whether domesticated stock, bred in captivity over tens of generations, differ in their response to a life-history manipulation, compared to birds taken directly from the wild. In a 'common aviary' experiment, we thereto experimentally manipulated the environmental conditions experienced by nestlings early in life by means of a brood size manipulation, and subsequently assessed its short- and long-term consequences on growth, ornamentation, immune function and reproduction. As expected, we found that early environmental conditions had a marked effect on both short- and long-term morphological and life-history traits in all birds. However, although there were pronounced differences between wild and domesticated birds with respect to the absolute expression of many of these traits, which are indicative of the different selection pressures wild and domesticated birds were exposed to in the recent past, manipulated rearing conditions affected morphology and ornamentation of wild and domesticated finches in a very similar way. This suggests that despite significant differentiation between wild and domesticated birds, selection has not altered the relationships among traits. Thus, life-history strategies and investment trade-offs may be relatively stable and not easily altered by selection. This is a reassuring finding in the light of the widespread use of domesticated birds in studies of life-history evolution and sexual selection, and suggests that adaptive explanations may be legitimate when referring to captive bird studies.

Environment-specific elasticity and sensitivity analysis of the stochastic growth rate

For matrix population models, analyses of how sensitive the population growth rate is to changes in vital rates (i.e. perturbations) are important for studies of life history evolution as well as for management and conservation of threatened species. There are two types of sensitivity analyses corresponding to absolute (sensitivity) or relative (elasticity) changes in the vital rates and both types can be applied to both deterministic and stochastic matrix population models. To date, most empirical studies of elasticity and sensitivity of the stochastic growth rate have examined the response to perturbations in the vital rates in a complete set of possible environments. However, it is often of interest to examine the response to perturbations occurring in only a subset of the possible environments. This has been done for periodic time-varying models elsewhere, but here we describe a recently published method for calculating the environment-specific sensitivity and elasticity of the stochastic growth rate and apply this method to data. These environment-specific perturbation analyses provide a logical way of dividing the sensitivity and elasticity among the environments. They give important insight into the selection regime in different environments and can provide valuable information for making management decisions and management evaluations in stochastic environments.

Individual variation in reproductive costs of reproduction: high‐quality females always do better

1. Although life-history theory predicts substantial costs of reproduction, individuals often show positive correlations among life-history traits, rather than trade-offs. The apparent absence of reproductive costs may result from heterogeneity in individual quality. 2. Using detailed longitudinal data from three contrasted ungulate populations (mountain goats, Oreamnos americanus; bighorn sheep, Ovis canadensis; and roe deer, Capreolus capreolus), we assessed how individual quality affects the probability of detecting a cost of current reproduction on future reproduction for females. We used a composite measure of individual quality based on variations in longevity (all species), success in the last breeding opportunity before death (goats and sheep), adult mass (all species), and social rank (goats only). 3. In all species, high-quality females consistently had a higher probability of reproduction, irrespective of previous reproductive status. In mountain goats, we detected a cost of reproduction only after accounting for differences in individual quality. Only low-quality female goats were less likely to reproduce following years of breeding than of nonbreeding. Offspring survival was lower in bighorn ewes following years of successful breeding than after years when no lamb was produced, but only for low-quality females, suggesting that a cost of reproduction only occurred for low-quality females. 4. Because costs of reproduction differ among females, studies of life-history evolution must account for heterogeneity in individual quality.

Phylogeny of the pollinating yucca moths, with revision of Mexican species (Tegeticula and Parategeticula; Lepidoptera, Prodoxidae)

ABSTRACT The yucca moths (Tegeticula and Parategeticula; Lepidoptera, Prodoxidae) are well-known for their obligate relationship as exclusive pollinators of yuccas. Revisionary work in recent years has revealed far higher species diversity than historically recognized, increasing the number of described species from four to 21. Based on field surveys in Mexico and examination of collections, we describe five additional species: T. californica Pellmyr sp. nov., T. tehuacana Pellmyr & Balcazar-Lara sp. nov., T. tambasi Pellmyr & Balcazar-Lara sp. nov., T. baja Pellmyr & Balcazar-Lara sp. nov., and P. californica Pellmyr & Balcazar-Lara sp. nov. Tegeticula treculeanella Pellmyr is identified as a junior synonym of T. mexicana Bastida. A diagnostic key to the adults of all species of the T. yuccasella complex is provided. A phylogeny based on a 2104-bp segment of mitochondrial DNA (mtDNA) in the cytochrome oxidase I and II region supported monophyly of the two pollinator genera, and strongly supported monophyly of the 17 recognized species of the T. yuccasella complex. Most relationships are well-supported, but some relationships within a recent and rapidly diversified group of 11 taxa are less robust, and in one case conflicts with a whole-genome data set (AFLP). The current mtDNA-based analyses, together with previouslymore » published AFLP data, provide a robust phylogenetic foundation for future studies of life history evolution and host interactions in one of the classical models of coevolution and obligate mutualism. ADDITIONAL KEY WORDS: mutualism, pollination, molecular phylogenetics, mitochondrial DNA« less

Sex‐specific costs of reproduction in Eastern Bluebirds Sialia sialis

In species with bi-parental care, individuals must partition energy between parental effort and mating effort. Typically, female songbirds invest more than males in reproductive activities such as egg-laying and incubation, but males invest more in secondary sexual traits used in attracting mates. Animals that breed more than once within a season must also allocate time and energy between first and subsequent breeding attempts and between current and future breeding seasons. To investigate strategies of reproductive investment by males and females and the consequences of such strategies, we manipulated the size of broods of Eastern Bluebirds Sialia sialis. Pairs with enlarged first broods were less likely to produce a second clutch or took longer to initiate one than pairs with reduced broods. After rearing enlarged broods, females were less likely than males to survive to the following year. Although plumage coloration is a sexually selected trait in Eastern Bluebirds that is influenced by nutritional stress, we did not detect an effect of brood-size manipulation on female coloration. Past research, however, demonstrates that, in males, plumage colour is negatively affected by increasing brood size. We suggest that there are sex-specific strategies of reproductive investment in Eastern Bluebirds, and that researchers should incorporate measures of residual reproductive value in studies of life-history evolution.

The Evolution of Senescence and Post-Reproductive Lifespan in Guppies (Poecilia reticulata)

The study of post-reproductive lifespan has been of interest primarily with regard to the extended post-menopausal lifespan seen in humans. This unusual feature of human demography has been hypothesized to have evolved because of the “grandmother” effect, or the contributions that post-reproductive females make to the fitness of their children and grandchildren. While some correlative analyses of human populations support this hypothesis, few formal, experimental studies have addressed the evolution of post-reproductive lifespan. As part of an ongoing study of life history evolution in guppies, we compared lifespans of individual guppies derived from populations that differ in their extrinsic mortality rates. Some of these populations co-occur with predators that increase mortality rate, whereas other nearby populations above barrier waterfalls are relatively free from predation. Theory predicts that such differences in extrinsic mortality will select for differences in the age at maturity, allocation of resources to reproduction, and patterns of senescence, including reproductive declines. As part of our evaluation of these predictions, we quantified differences among populations in post-reproductive lifespan. We present here the first formal, comparative study of the evolution of post-reproductive lifespan as a component of the evolution of the entire life history.Guppies that evolved with predators and that experienced high extrinsic mortality mature at an earlier age but also have longer lifespans. We divided the lifespan into three non-overlapping components: birth to age at first reproduction, age at first reproduction to age at last reproduction (reproductive lifespan), and age at last reproduction to age at death (post-reproductive lifespan). Guppies from high-predation environments live longer because they have a longer reproductive lifespan, which is the component of the life history that can make a direct contribution to individual fitness. We found no differences among populations in post-reproductive lifespan, which is as predicted since there can be no contribution of this segment of the life history to an individual's fitness.Prior work on the evolution of post-reproductive lifespan has been dominated by speculation and correlative analyses. We show here that this component of the life history is accessible to formal study as part of experiments that quantify the different segments of an individual's life history. Populations of guppies subject to different mortality pressures from predation evolved differences in total lifespan, but not in post-reproductive lifespan. Rather than showing the direct effects of selection characterizing other life-history traits, post-reproductive lifespan in these fish appears to be a random add-on at the end of the life history. These findings support the hypothesis that differences in lifespan evolving in response to selection are confined to the reproductive lifespan, or those segments of the life history that make a direct contribution to fitness. We also show, for the first time, that fish can have reproductive senescence and extended post-reproductive lifespans despite the general observation that they are capable of producing new primary oocytes throughout their lives.

Carotenoids modulate the trade-off between egg production and resistance to oxidative stress in zebra finches

The allocation of resources to reproduction and survival is a central question of studies of life history evolution. Usually, increased allocation to current reproduction is paid in terms of reduced future reproduction and/or decreased survival. However, the proximal mechanisms underlying the cost of reproduction are poorly understood. Recently, it has been shown that increased susceptibility to oxidative stress might be one of such proximate links between reproduction and self-maintenance. Organisms possess a range of antioxidant defenses, including endogenously produced molecules (e.g., enzymes) and compounds ingested with food (e.g., carotenoids). If reproductive effort increases the production of reactive oxygen species, the availability of antioxidant defenses may partly or fully counteract the free-radical damages. One could, therefore, expect that the trade-off between reproduction and oxidative stress is modulated by the availability of antioxidant defenses. We tested this hypothesis in zebra finches. We manipulated reproductive effort by either allowing or preventing pairs to breed. Within each breeding or non-breeding group, the availability of antioxidant compounds was manipulated by supplementing or not supplementing the drinking water with carotenoids. We found that although birds in the breeding and non-breeding groups did not differ in their resistance to oxidative stress (the breakdown of red blood cells submitted to a controlled free-radical attack), one aspect of breeding effort (i.e., the number of eggs laid by birds in both breeding and non-breeding groups) was negatively correlated with resistance to oxidative stress only in birds that did not benefit from a carotenoid-supplemented diet. This result therefore suggests that carotenoid availability can modulate the trade-off between reproduction and resistance to oxidative stress.

Inter‐population variation in demographic parameters: a neglected subject?

Demographic parameters and population growth rates vary among populations of the same species. Such variation can be extensive, as shown by a comparison of nine published studies of black‐legged kittiwakes Rissa tridactyla. We argue that rather than being a nuisance, inter‐population variation can improve our understanding of population dynamics. Analysis of data from several geographically dispersed studies can aid detection of causal relationships between demographic parameters and environmental factors and thus improve our understanding of impacts of e.g. climate change, and help elucidate mechanisms of population regulation. Extensive inter‐population variation also adds another perspective to studies of life history evolution and should be taken into account in across‐species comparisons.

The evolution of alternative parasitic life histories in large blue butterflies

Large blue (Maculinea) butterflies are highly endangered throughout the Palaearctic region, and have been the focus of intense conservation research. In addition, their extraordinary parasitic lifestyles make them ideal for studies of life history evolution. Early instars consume flower buds of specific host plants, but later instars live in ant nests where they either devour the brood (predators), or are fed mouth-to-mouth by the adult ants (cuckoos). Here we present the phylogeny for the group, which shows that it is a monophyletic clade nested within Phengaris, a rare Oriental genus whose species have similar life histories. Cuckoo species are likely to have evolved from predatory ancestors. As early as five million years ago, two Maculinea clades diverged, leading to the different parasitic strategies seen in the genus today. Contrary to current belief, the two recognized cuckoo species show little genetic divergence and are probably a single ecologically differentiated species. On the other hand, some of the predatory morphospecies exhibit considerable genetic divergence and may contain cryptic species. These findings have important implications for conservation and reintroduction efforts.