Asynchrony, density dependence, and persistence in an amphibian.

Understanding drivers of metapopulation dynamics remains a critical challenge for ecology and conservation. In particular, the degree of synchrony in metapopulation dynamics determines how resilient a metapopulation is to a widespread disturbance. In this study, we used 21 years of egg mass count data across 64 nonpermanent freshwater ponds in Connecticut, USA to evaluate patterns of abundance and growth and to assess regional as well as local factors in shaping the population dynamics of wood frogs (Rana sylvatica = Lithobates sylvaticus). In particular, we asked whether a species known to undergo metapopulation dynamics exhibited spatial synchrony in abundances. With the exception of a single year when breeding took place during severe drought conditions, our analyses revealed no evidence of synchrony despite close proximity (mean minimum distance < 300 m) of breeding ponds across the 3213-ha study area. Instead, local, pond-scale conditions best predicted patterns of abundance and population growth rate. We found negative density dependence on population growth rate within ponds as well as evidence that larger neighboring pond populations had a negative effect on focal ponds. Beyond density, pond depth was a critical predictor; deeper ponds supported larger populations. Drought conditions and warm winters negatively affected populations. Overall, breeding ponds vary in critical ways that either support larger, more persistent populations or smaller populations that are not represented by breeding pairs in some years. The infrequency of spatial synchrony in this system is surprising and suggests greater resilience to stressors than would have been expected if dynamics were strongly synchronized. More generally, understanding the characteristics of systems that determine synchronous population dynamics will be critical to predicting which species are more or less resilient to widespread disturbances like land conversion or climate change.

Rapid microgeographic evolution in response to climate change.

Environmental change is predicted to accelerate into the future and will exert strong selection pressure on biota. Although many species may be fated to extinction, others may survive through their capacity to evolve rapidly at highly localized (i.e., microgeographic) scales. Yet, even as new examples have been discovered, the limits to such evolutionary responses have not often been evaluated. One of the first examples of microgeographic variation involved pond populations of wood frogs (Rana sylvatica). Although separated by just tens to hundreds of meters, these populations exhibited countergradient variation in intrinsic embryonic development rates when reared in a common garden. We repeated this experiment 17 years (approximately six to nine generations) later and found that microgeographic variation persists in contemporary populations. Furthermore, we found that contemporary embryos have evolved to develop 14-19% faster than those in 2001. Structural equation models indicate that the predominant cause for this response is likely due to changes in climate over the intervening 17 years. Despite potential for rapid and fine-scale evolution, demographic declines in populations experiencing the greatest changes in climate and habitat imply a limit to the species' ability to mitigate extreme environmental change.

Metamorphosis in an Era of Increasing Climate Variability.

Most animals have complex life cycles including metamorphosis or other discrete life stage transitions, during which individuals may be particularly vulnerable to environmental stressors. With climate change, individuals will be exposed to increasing thermal and hydrologic variability during metamorphosis, which may affect survival and performance through physiological, behavioral, and ecological mechanisms. Furthermore, because metamorphosis entails changes in traits and vital rates, it is likely to play an important role in how populations respond to increasing climate variability. To identify mechanisms underlying population responses and associated trait and life history evolution, we need new approaches to estimating changes in individual traits and performance throughout metamorphosis, and we need to integrate metamorphosis as an explicit life stage in analytical models.

Temperature-mediated trade-off between development and performance in larval wood frogs (Rana sylvatica).

Countergradient variation has been detected in diverse taxa. In a common manifestation, individuals from colder environments develop faster than conspecifics from warmer environments when placed in a common garden. Where such a pattern exists, it implies a trade-off: Individuals from warmer environments have intrinsic rates of development lower than those demonstrated by other individuals of the same species. We explored a trade-off between development rate and locomotor performance in the wood frog (Rana sylvatica), an amphibian for which countergradient variation has been well documented. We reared wood frogs from 10 populations under two temperature regimes, bracketing the temperatures observed in local natural ponds. Individuals reared under warmer conditions developed more rapidly but exhibited burst speeds 20% lower than individuals reared under colder conditions. The slope of the reaction norm was consistent across the 10 populations and thus, we found no evidence of countergradient variation in performance. Burst speed assays of wild-caught tadpoles from the same populations showed a similar but nonsignificant trend, with greater variability among ponds. Overall, our findings support the existence of a development-performance trade-off that may be of broad importance and which may help explain the widespread occurrence of countergradient variation.

Suburbanization Increases Echinostome Infection in Green Frogs and Snails.

An important contribution to infectious disease emergence in wildlife is environmental degradation driven by pollution, habitat fragmentation, and eutrophication. Amphibians are a wildlife group that is particularly sensitive to land use change, infectious diseases, and their interactions. Residential suburban land use is now a dominant, and increasing, form of land cover in the USA and globally, contributing to increased pollutant and nutrient loading in freshwater systems. We examined how suburbanization affects the infection of green frog (Rana clamitans) tadpoles and metamorphs by parasitic flatworms (Echinostoma spp.) through the alteration of landscapes surrounding ponds and concomitant changes in water quality. Using sixteen small ponds along a forest-suburban land use gradient, we assessed how the extent of suburban land use surrounding ponds influenced echinostome infection in both primary snail and secondary frog hosts. Our results show that the degree of suburbanization and concurrent chemical loading are positively associated with the presence and burden of echinostome infection in both host populations. This work contributes to a broader understanding of how land use mediates wildlife parasitism and shows how human activities at the household scale can have similar consequences for wildlife health as seemingly more intensive land uses like agriculture or urbanization.

Molecular evidence for sex reversal in wild populations of green frogs (Rana clamitans).

In vertebrates, sex determination occurs along a continuum from strictly genotypic (GSD), where sex is entirely guided by genes, to strictly environmental (ESD), where rearing conditions, like temperature, determine phenotypic sex. Along this continuum are taxa which have combined genetic and environmental contributions to sex determination (GSD + EE), where some individuals experience environmental effects which cause them to sex reverse and develop their phenotypic sex opposite their genotypic sex. Amphibians are often assumed to be strictly GSD with sex reversal typically considered abnormal. Despite calls to understand the relative natural and anthropogenic causes of amphibian sex reversal, sex reversal has not been closely studied across populations of any wild amphibian, particularly in contrasting environmental conditions. Here, we use sex-linked molecular markers to discover sex reversal in wild populations of green frogs (Rana clamitans) inhabiting ponds in either undeveloped, forested landscapes or in suburban neighborhoods. Our work here begins to suggest that sex reversal may be common within and across green frog populations, occurring in 12 of 16 populations and with frequencies of 2-16% of individuals sampled within populations. Additionally, our results also suggest that intersex phenotypic males and sex reversal are not correlated with each other and are also not correlated with suburban land use. While sex reversal and intersex are often considered aberrant responses to human activities and associated pollution, we found no such associations here. Our data perhaps begin to suggest that, relative to what is often suggested, sex reversal may be a relatively natural process in amphibians. Future research should focus on assessing interactions between genes and the environment to understand the molecular and exogenous basis of sex determination in green frogs and in other amphibians.

Sexual and somatic development of wood frog tadpoles along a thermal gradient.

All amphibian species are known to have genetic sex determination. However, a variety of environmental conditions can moderate sexual differentiation, in some cases leading to sex reversal and skewed sex ratios. While there has been a recent focus on chemically-induced sex reversal in amphibians, temperature can also influence sexual differentiation. Building upon a classic 1929 study by Emil Witschi, we assessed temperature-mediated sex reversal. Witschi found that the wood frog sex ratio is 100% male at a high temperature (32°C) compared to a 50:50 sex ratio at 20°C. This pattern is consistent with multiple models of environmentally mediated sexual differentiation in vertebrates. To better understand thermally mediated sex reversal, we raised wood frogs at temperature increments of ∼1°C between 19 and 34°C. Mirroring earlier findings, wood frog metamorph sex ratios are indistinguishable from 50:50 at the lowest temperature and entirely male at the highest temperatures. In between, sex ratios become increasingly male-dominated as temperatures increase, implying a steadily increasing tendency toward female-to-male sex reversal in warmer environments. There was no evidence of a threshold temperature effect on reversal patterns. We also show that, compared to males, females metamorphose larger and later in cooler conditions but earlier and smaller under warmer conditions. While the ecological relevance in this species is unknown, these results conform to the Charnov-Bull model of sex determination (in which female-to-male sex reversal can increase fitness to genetic females at higher temperatures), suggesting the system would reward further study.

Range position and climate sensitivity: The structure of among-population demographic responses to climatic variation.

Species' distributions will respond to climate change based on the relationship between local demographic processes and climate and how this relationship varies based on range position. A rarely tested demographic prediction is that populations at the extremes of a species' climate envelope (e.g., populations in areas with the highest mean annual temperature) will be most sensitive to local shifts in climate (i.e., warming). We tested this prediction using a dynamic species distribution model linking demographic rates to variation in temperature and precipitation for wood frogs (Lithobates sylvaticus) in North America. Using long-term monitoring data from 746 populations in 27 study areas, we determined how climatic variation affected population growth rates and how these relationships varied with respect to long-term climate. Some models supported the predicted pattern, with negative effects of extreme summer temperatures in hotter areas and positive effects on recruitment for summer water availability in drier areas. We also found evidence of interacting temperature and precipitation influencing population size, such as extreme heat having less of a negative effect in wetter areas. Other results were contrary to predictions, such as positive effects of summer water availability in wetter parts of the range and positive responses to winter warming especially in milder areas. In general, we found wood frogs were more sensitive to changes in temperature or temperature interacting with precipitation than to changes in precipitation alone. Our results suggest that sensitivity to changes in climate cannot be predicted simply by knowing locations within the species' climate envelope. Many climate processes did not affect population growth rates in the predicted direction based on range position. Processes such as species-interactions, local adaptation, and interactions with the physical landscape likely affect the responses we observed. Our work highlights the need to measure demographic responses to changing climate.

Searching for Biotic Multipliers of Climate Change.

SYNOPSIS: As climates change, biologists need to prioritize which species to understand, predict, and protect. One way is to identify key species that are both sensitive to climate change and that disproportionately affect communities and ecosystems. These "biotic multipliers" provide efficient targets for research and conservation. Here, we propose eight mechanistic hypotheses related to impact and sensitivity that suggest that top consumers might often act as biotic multipliers of climate change. For impact, top consumers often affect communities and ecosystems through strong top-down effects. For sensitivity, metabolic theory and data suggest that photosynthesis and respiration differ in temperature responses, potentially increasing the sensitivity of consumers relative to plants. Larger-bodied organisms are typically more thermally sensitive than smaller ones, suggesting how large top consumers might be more sensitive than their smaller prey. In addition, traits related to predation are more sensitive than defensive traits to temperature. Top consumers might also be more sensitive because they often lag behind prey in phenological responses. The combination of low population sizes and demographic traits of top consumers could make them more sensitive to disturbances like climate change, which could slow their recovery. As top consumers are positioned at the top of the food chain, many small effects can accumulate from other trophic levels to affect top consumers. Finally, top consumers also often disperse more frequently and farther than prey, potentially leading to greater sensitivity to climate-induced changes in ranges and subsequent impacts on invaded communities. Overall, we expect that large, ectothermic top consumers and mobile predators might frequently be biotic multipliers of climate change. However, this prediction depends on the particular features of species, habitats, and ecosystems. In specific cases, herbivores, plants, or pathogens might be more sensitive than top consumers or have greater community impacts. To predict biotic multipliers, we need to compare sensitivities and impacts across trophic groups in a broader range of ecosystems as well as perform experiments that uncouple proposed mechanisms. Overall, the biotic multiplier concept offers an alternative prioritization scheme for research and conservation that includes impacts on communities and ecosystems.

Daphniid zooplankton assemblage shifts in response to eutrophication and metal contamination during the Anthropocene.

Human activities during the Anthropocene result in habitat degradation that has been associated with biodiversity loss and taxonomic homogenization of ecological communities. Here we estimated effects of eutrophication and heavy metal contamination, separately and in combination, in explaining zooplankton species composition during the past 125-145 years using analysis of daphniid diapausing egg banks from four lakes in the northeastern USA. We then examined how these community shifts influenced patterns of diversity and homogenization. Analysis of past lake production (via subfossil pigments) and metal contamination (via sedimentary metals) demonstrated that eutrophication alone (19-39%) and in combination with metal pollution (17-54%) explained 36-79% of historical variation in daphniid species relative abundances in heavily fertilized lakes. In contrast, metal pollution alone explained the majority (72%) of historical variation in daphniid assemblages at the oligotrophic site. Several species colonization events in eutrophying lakes resulted in increased species richness and gamma diversity through time. At the same time, daphniid assemblages in three eutrophied lakes became more similar to each other (homogenized), but this pattern was only seen when accounting for species presence/absence. We did not observe consistent patterns of divergence between the assemblages in the eutrophying lakes and the low-nutrient reference site. Given the pervasive nature of fertilization and metal pollution, and the sensitivity of cladocerans to these factors, we suggest that many inhabited lake districts may already exhibit similar patterns of daphniid assemblage shifts.

Reconciling the role of terrestrial leaves in pond food webs: a whole-ecosystem experiment.

Terrestrial carbon and nutrients can subsidize the detrital pool of freshwater ecosystems; yet, the importance of terrestrial subsidies to lake and pond food webs is uncertain and debated. Terrestrial detritus is expected to have the greatest impact on food webs when water bodies are small and shallow with low levels of incident light. Temporary forested ponds fit this description and are often assumed to have a leaf detritus-based food web, but this has not been quantified. In a whole-ecosystem experiment, we traced the flow of isotopically enriched leaf litter to primary producers and consumers in a small, forested pond. We found that terrestrial leaves provided nutrients to algae, offering an indirect pathway in which leaf litter can enter the food web. Terrestrial leaves were also consumed directly, and larval caddisfly (Limnephilus sp.) shredders likely mobilized leaf nutrients to other consumers, a process overlooked in many previous small-scale experiments that did not incorporate shredders. Unexpectedly, most consumers relied heavily upon algal food pathways despite low light and net heterotrophic conditions. Overall, our study highlights the interconnectedness of algal and leaf litter pathways in small pond food webs, and emphasizes that algal pathways are prevalent and important even in small, shaded ponds with high loads of terrestrial leaf litter.

Sex-linked markers in the North American green frog (Rana clamitans) developed using DArTseq provide early insight into sex chromosome evolution.

BACKGROUND: The extent to which sex reversal is associated with transitions in sex determining systems (XX-XY, ZZ-ZW, etc.) or abnormal sexual differentiation is predominantly unexplored in amphibians. This is in large part because most amphibian taxa have homomorphic sex chromosomes, which has traditionally made it challenging to identify discordance between phenotypic and genetic sex in amphibians, despite all amphibians having a genetic component to sex determination. Recent advances in molecular techniques such as genome complexity reduction and high throughput sequencing present a valuable avenue for furthering our understanding of sex determination in amphibians and other taxa with homomorphic sex chromosomes like many fish and reptiles. RESULTS: We use DArTseq as a novel approach to identify sex-linked markers in the North American green frog (Rana clamitans melanota) using lab-reared tadpoles as well as wild-caught adults from seven ponds either in undeveloped, forested habitats or suburban ponds known to be subject to contamination by anthropogenic chemicals. The DArTseq methodology identified 13 sex-linked SNP loci and eight presence-absence loci associated with males, indicating an XX-XY system. Both alleles from a single locus show partial high sequence homology to Dmrt1, a gene linked to sex determination and differentiation throughout Metazoa. Two other loci have sequence similarities to regions of the chimpanzee and human X-chromosome as well as the chicken Z-chromosome. Several loci also show geographic variation in sex-linkage, possibly indicating sex chromosome recombination. While all loci are statistically sex-linked, they show varying degrees of female heterozygosity and male homozygosity, providing further evidence that some markers are on regions of the sex chromosomes undergoing higher rates of recombination and therefore further apart from the putative sex determining locus. CONCLUSION: The ease of the DArTseq platform provides a useful avenue for future research on sex reversal and sex chromosome evolution in vertebrates, particularly for non-model species with homomorphic or cryptic or nascent sex chromosomes.

Septic systems, but not sanitary sewer lines, are associated with elevated estradiol in male frog metamorphs from suburban ponds.

Suburban neighborhoods are a dominant type of human land use. Many housing regions globally rely on septic systems, rather than sanitary sewers, for wastewater management. There is evidence that septic systems may contaminate waterbodies more than sewer lines. There is also mounting evidence that human activities contaminate waterways with endocrine-disrupting chemicals (EDCs), which alter wildlife sexual development. While endocrine disruption is often associated with intense activities such as agriculture or wastewater treatment plant discharges, recent evidence indicates that endocrine disruption is pervasive in frogs from suburban neighborhoods. In conjunction with other putative EDC sources, one hypothesis is that wastewater is contaminating suburban waterways with EDCs derived from pharmaceuticals or personal care products. Here, we measure estradiol (E2) in metamorphosing green frogs (Rana clamitans) from forested ponds and suburban ponds adjacent to either septic tanks or sanitary sewers. We show that E2 is highest in male frogs from septic neighborhoods and that E2 concentrations are significantly lower in male frogs from forested ponds and from ponds near sewers. These results indicate that septic tanks may be contaminating aquatic ecosystems differently than sewer lines. This pattern contrasts prior work showing no difference in EDC contamination or morphological endocrine disruption between septic and sewer neighborhoods, implying that suburbanization may have varying effects at multiple biological scales like physiology and anatomy.

Suburbanization, estrogen contamination, and sex ratio in wild amphibian populations.

Research on endocrine disruption in frog populations, such as shifts in sex ratios and feminization of males, has predominantly focused on agricultural pesticides. Recent evidence suggests that suburban landscapes harbor amphibian populations exhibiting similar levels of endocrine disruption; however the endocrine disrupting chemical (EDC) sources are unknown. Here, we show that sex ratios of metamorphosing frogs become increasingly female-dominated along a suburbanization gradient. We further show that suburban ponds are frequently contaminated by the classical estrogen estrone and a variety of EDCs produced by plants (phytoestrogens), and that the diversity of organic EDCs is correlated with the extent of developed land use and cultivated lawn and gardens around a pond. Our work also raises the possibility that trace-element contamination associated with human land use around suburban ponds may be contributing to the estrogenic load within suburban freshwaters and constitutes another source of estrogenic exposure for wildlife. These data suggest novel, unexplored pathways of EDC contamination in human-altered environments. In particular, we propose that vegetation changes associated with suburban neighborhoods (e.g., from forests to lawns and ornamental plants) increase the distribution of phytoestrogens in surface waters. The result of frog sex ratios varying as a function of human land use implicates a role for environmental modulation of sexual differentiation in amphibians, which are assumed to only have genetic sex determination. Overall, we show that endocrine disruption is widespread in suburban frog populations and that the causes are likely diverse.

Cross-scale interactions and the distribution-abundance relationship.

Positive interspecific relationships between local abundance and extent of regional distribution are among the most ubiquitous patterns in ecology. Although multiple hypotheses have been proposed, the mechanisms underlying distribution-abundance (d-a) relationships remain poorly understood. We examined the intra- and interspecific distribution-abundance relationships for a metacommunity of 13 amphibian species sampled for 15 consecutive years. Mean density of larvae in occupied ponds was positively related to number of ponds occupied by species; employing the fraction of ponds uniquely available to each species this same relationship sharply decelerates. The latter relationship suggested that more abundant species inhabited most available habitats annually, whereas rarer species were dispersal limited. We inferred the mechanisms responsible for this pattern based on the dynamics of one species, Pseudacris triseriata, which transitioned between a rare, narrowly distributed species to a common, widely distributed species and then back again. Both transitions were presaged by marked changes in mean local densities driven by climatic effects on habitat quality. We identified threshold densities separating these population regime shifts that differed with landscape configuration. Our data suggest that these transitions were caused by strong cross-scale interactions between local resource/niche processes and larger scale metapopulation processes. The patterns we observed have relevance for understanding the mechanisms of interspecific d-a relationships and critical thresholds associated with habitat fragmentation.

Experimental canopy removal enhances diversity of vernal pond amphibians.

Vernal ponds are often treated as protected environments receiving special regulation and management. Within the landscapes where they are found, forest vegetation frequently dominates surrounding uplands and can grow to overtop and shade pond basins. Two bodies of research offer differing views of the role of forest canopy for vernal pond systems. Studies of landscape conversion suggest that removing forest overstory within uplands can cause local extinctions of amphibians by altering terrestrial habitat or hindering movement. Studies of canopy above pond basins imply an opposite relationship; encroachment of overstory vegetation can be associated with local extinctions potentially via changes in light, thermal, and food resource environments. Unresolved uncertainties about the role of forest canopy reveal significant gaps in our understanding of wetland species distributions and dynamics. Any misunderstanding of canopy influences is simultaneously important to managers because current practices emphasize promoting or conserving vegetation growth particularly within buffers immediately adjacent to ponds. We evaluated this apparent contradiction by conducting a landscape-scale, long-term experiment using 14 natural vernal ponds. Tree felling at six manipulated ponds was limited in spatial scope but was nevertheless effective in increasing water temperature. Compared with eight control ponds, manipulated ponds maintained more amphibian species during five years post-manipulation. There was little evidence that any species was negatively influenced, and the reproductive effort of species for which we estimated egg inputs maintained pretreatment population densities in manipulated compared with control ponds. Overall, our experiment shows that a carefully circumscribed reduction of overhead forest canopy can enhance the capacity of vernal ponds to support wildlife diversity and suggests a scale dependence of canopy influences on amphibians. These findings have implications for the connection between current wetland management practices and the goals of wetland stewardship and conservation of wetland-dependent species.

Microgeographic adaptation and the spatial scale of evolution.

Local adaptation has been a major focus of evolutionary ecologists working across diverse systems for decades. However, little of this research has explored variation at microgeographic scales because it has often been assumed that high rates of gene flow will prevent adaptive divergence at fine spatial scales. Here, we establish a quantitative definition of microgeographic adaptation based on Wright's dispersal neighborhood that standardizes dispersal abilities, enabling this measure to be compared across species. We use this definition to evaluate growing evidence of evolutionary divergence at fine spatial scales. We identify the main mechanisms known to facilitate this adaptation and highlight illustrative examples of microgeographic evolution in nature. Collectively, this evidence requires that we revisit our understanding of the spatial scale of adaptation and consider how microgeographic adaptation and its driving mechanisms can fundamentally alter ecological and evolutionary dynamics in nature.

Moving forward: dispersal and species interactions determine biotic responses to climate change.

We need accurate predictions about how climate change will alter species distributions and abundances around the world. Most predictions assume simplistic dispersal scenarios and ignore biotic interactions. We argue for incorporating the complexities of dispersal and species interactions. Range expansions depend not just on mean dispersal, but also on the shape of the dispersal kernel and the population's growth rate. We show how models using species-specific dispersal can produce more accurate predictions than models applying all-or-nothing dispersal scenarios. Models that additionally include species interactions can generate distinct outcomes. For example, species interactions can slow climate tracking and produce more extinctions than models assuming no interactions. We conclude that (1) just knowing mean dispersal is insufficient to predict biotic responses to climate change, and (2) considering interspecific dispersal variation and species interactions jointly will be necessary to anticipate future changes to biological diversity. We advocate for collecting key information on interspecific dispersal differences and strong biotic interactions so that we can build the more robust predictive models that will be necessary to inform conservation efforts as climates continue to change.