Ecological and Fitness Correlates of Personality in a Long-lived Terrestrial Turtle.

An individual's behavioral tendencies (i.e., personality or temperament) can influence its interactions with the environment and thus have important ecological and evolutionary consequences for animal populations. Boldness, defined as an individual's tendency to engage in risk-taking activities, is a phenotypically variable trait linked with numerous behavioral and fitness outcomes in free-ranging animals. We examined variation and repeatability of boldness and other behavioral characteristics in two wild Eastern Box Turtle (Terrapene carolina carolina) populations using radiotelemetry, and assessed fitness correlates of boldness over multiple years. We observed large amounts of among-individual variation and within-individual consistency (i.e., repeatability) of boldness as measured by their head emergence latency following a standardized confinement assay. Individuals were also consistent in several in-field behaviors including movement rate, home range size, and date of emergence from overwintering refuges. Individuals with shorter head emergence latencies (i.e., bolder turtles) had larger home ranges, emerged earlier from overwintering dormancy, and experienced moderately lower survival compared with shy individuals. Boldness did not affect time spent within the thermal preference range, somatic growth rates, or the frequency of mating or same-sex aggressive encounters. Boldness and its effects on in-field behaviors differed between sexes and populations, and the relationship between boldness and survival was temporally variable. Our results suggest possible intrinsic behavioral types in T. c. carolina and highlight the importance of long-term and multipopulation studies when examining ecological and evolutionary processes that shape personality phenotypes in turtles.

Spatial Ecology, Movements, and Habitat Selection of Clemmys guttata in a Temporally Dynamic Wetland System in North Carolina, USA.

Turtle populations are in decline worldwide, requiring immediate conservation and management actions. For species with broad geographic ranges that cover diverse environmental contexts, region-specific information on declining species could inform more targeted management plans. I examined the ecology of a Spotted Turtle (Clemmys guttata) population in a temporally dynamic wetland system in the Southeastern Plains ecoregion of North Carolina. Turtles selected forested wetlands and streams while avoiding open wetlands and river habitats, but used terrestrial habitats minimally and randomly. Turtles responded to wetland drying by remaining in wetlands and maintaining modest levels of activity during short-duration drying events (< 1 mo), but moved very little during longer droughts (7 mo). Turtles had prolonged active seasons (February-October) as long as wetlands and streams were flooded, with movement rates peaking in late spring at 23.6 ± 3.7 m/d (mean ± standard error). Turtles had large home ranges (14.1 ± 4.3 ha) that often included multiple local stream networks used as movement corridors between forested wetland patches and as activity centers when wetlands dried. I suggest that conservation plans for C. guttata include large management areas that protect a network of streams and adjacent forested swamps in the Southeastern Plains and perhaps other lowland ecoregions in the Southeastern U.S. Because terrestrial habitats were not used for extended refuge during drought, certain low-impact land uses in adjacent terrestrial areas are not likely to negatively impact C. guttata populations, but terrestrial forests would still be important in maintaining overland travel corridors.

Metabolic consequences of sex reversal in two lizard species: a test of the like-genotype and like-phenotype hypotheses.

Vertebrate sex is typically determined genetically, but in many ectotherms sex can be determined by genes (genetic sex determination, GSD), temperature (temperature-dependent sex determination, TSD), or interactions between genes and temperature during development. TSD may involve GSD systems with either male or female heterogamety (XX/XY or ZZ/ZW) where temperature overrides chromosomal sex determination to cause a mismatch between genetic sex and phenotypic sex (sex reversal). In these temperature-sensitive lineages, phylogenetic investigations point to recurrent evolutionary shifts between genotypic and temperature-dependent sex determination. These evolutionary transitions in sex determination can occur rapidly if selection favours the reversed sex over the concordant phenotypic sex. To investigate the consequences of sex reversal on offspring phenotypes, we measured two energy-driven traits (metabolism and growth) and 6 month survival in two species of reptile with different patterns of temperature-induced sex reversal. Male sex reversal occurs in Bassiana duperreyi when chromosomal females (female XX) develop male phenotypes (maleSR XX), while female sex reversal occurs in Pogona vitticeps when chromosomal males (male ZZ) develop female phenotypes (femaleSR ZZ). We show metabolism in maleSR XX was like that of male XY; that is, reflective of phenotypic sex and lower than genotypic sex. In contrast, for Pogona vitticeps, femaleSR ZZ metabolism was intermediate between male ZZ and female ZW metabolic rate. For both species, our data indicate that differences in metabolism become more apparent as individuals become larger. Our findings provide some evidence for an energetic advantage from sex reversal in both species but do not exclude energetic processes as a constraint on the distribution of sex reversal in nature.

Evolutionary stability inferred for a free ranging lizard with sex-reversal.

The sex of vertebrates is typically determined genetically, but reptile sex can also be determined by developmental temperature. In some reptiles, temperature interacts with genotype to reverse sex, potentially leading to transitions from a chromosomal to a temperature-dependent sex determining system. Transitions between such systems in nature are accelerated depending on the frequency and fitness of sex-reversed individuals. The Central Bearded Dragon, Pogona vitticeps, exhibits female heterogamety (ZZ/ZW) but can have its sex reversed from ZZ male to ZZ female by high incubation temperatures. The species exhibits sex-reversal in the wild and it has been suggested that climate change and fitness of sex-reversed individuals could be increasing the frequency of reversal within the species range. Transitions to temperature-dependent sex determination require low levels of dispersal and high (>50%) rates of sex-reversal. Here, we combine genotype-by-sequencing, identification of phenotypic and chromosomal sex, exhaustive field surveys, and radio telemetry to examine levels of genetic structure, rates of sex-reversal, movement, space use, and survival of P. vitticeps in a location previously identified as a hot spot for sex-reversal. We find that the species exhibits low levels of population structure (FST ~0.001) and a modest (~17%) rate of sex-reversal, and that sex-reversed and nonsex-reversed females have similar survival and behavioural characteristics to each other. Overall, our data indicate this system is evolutionary stable, although we do not rule out the prospect of a more gradual transition in sex-determining mechanisms in the future in a more fragmented landscape and as global temperatures increase.

Overwintering behavior reduces mortality for a terrestrial turtle in forests managed with prescribed fire.

Prescribed fire is an essential management practice in pyrogenic ecosystems, but fire can also be a significant disturbance and source of mortality for both target and non-target species. Seasonal periods of animal inactivity may provide opportunities to design burn plans that minimize negative impacts to species of conservation concern, but few studies have rigorously examined this possibility. Using radiotelemetry, we studied overwintering behavior and interactions with fire in a forest-dwelling terrestrial turtle, the Eastern Box Turtle (Terrapene carolina carolina), over an eight-year period at two sites that use prescribed fire in forest management. Turtles at both sites selected predominantly hardwood forests and mesic habitats and avoided upland pine forests. Turtles buried deepest (2.9 - 3.2 cm) below the soil-litter interface in late February and then moved gradually shallower until emergence in early April. Emergence timing varied over a 58-day period, but was consistent within individuals from year to year. Turtles also maintained fidelity to refuge locations, but those overwintering in burned areas selected sites over twice as far from refuges used in previous years compared to those in unburned areas. The areas and habitats selected by turtles during winter served as refugia from fire, and those whose refuges did burn remained buffered from lethal temperatures even at shallow burial depths. The only fire-related injury or mortality occurred during seasons of surface activity. Timing burning and other forest management practices during periods of winter dormancy may thus minimize threats to turtle populations, but modifications to prescribed fire regimes must also be balanced with other management objectives.

Responses of a forest-dwelling terrestrial turtle, Terrapene Carolina, to prescribed fire in a Longleaf Pine ecosystem.

Prescribed fire is commonly used as a tool to meet a range of forest management goals. Owing to their limited movement abilities, terrestrial turtles are likely to be at high risk of injury and mortality, and to experience other fitness consequences with population-level implications from fire. Using radiotelemetry, we studied the responses of Eastern Box Turtles, Terrapene carolina carolina, to prescribed fire management in a sandhills Longleaf Pine forest system over a five-year period and compared our results to a nearby population in an unburned coastal plain location. Individual variation in turtle survival was strongly dependent on how frequently and extensively the areas were burned, with annual survival rates of 94.5% in unburned areas decreasing to 45.9 % in the most extensively burned areas. Turtles at the fire-maintained sandhills site had annual survival rates 4.9 % less than at the unburned coastal plain site, and females had annual survival rates 6.8 % less than males. Survival varied seasonally, with greatest mortality rates in winter and spring, especially among females. Growth rates and body condition did not differ between sites, nor did they vary according to fire extent and frequency at the fire-maintained site. Although mortality was greater and spatially variable at the fire-maintained site, annual survival rates across the site (86 - 90 % for females and males, respectively) were comparable to other stable populations of T. carolina. The lesser than expected mortality rate at the fire-maintained site was likely the result of turtles' strong selection of mesic hardwood forests near bottomlands and streams - habitats that may serve as refugia from fire. In areas where T. carolina conservation is a priority, land managers should integrate maintenance of fire refuge habitats into burn planning to minimize unintended negative impacts to this imperiled species.

Inter- and Intra-population Variation in Habitat Selection for a Forest-dwelling Terrestrial Turtle, Terrapene carolina carolina.

Habitat selection, where observed use of a resource is disproportionate to availability, is an important behavior allowing individuals to position themselves spatially relative to critical resources in heterogeneous environments. For species that experience variable environments across broad geographic ranges, we expect resource selection templates to vary among populations accordingly. Using radiotelemetry, we examined habitat selection for populations of Eastern Box Turtles, Terrapene carolina, in fire-maintained forests of the sandhills compared to nearby unburned coastal plain forests in south-central North Carolina. Turtles at the fire-maintained sandhills site preferred bottomland habitats and areas near steams, whereas turtles in the unburned coastal plain environment preferred uplands and used streams randomly. In addition, turtles in the fire-maintained sandhills avoided Longleaf Pine and more strongly preferred hardwood and non-Longleaf Pine forests compared to turtles at the unburned coastal plain site. Body size, but not sex, was also an important source of variation in habitat selection within populations, with smaller turtles more strongly preferring areas near water. Selection of habitat structural components in the immediate area of locations did not differ between sites, sexes, or body sizes. These results highlight the variety of resource selection templates in T. carolina, underscoring a potential need for population- or region-specific conservation and management strategies.

Thermal biology of eastern box turtles in a longleaf pine system managed with prescribed fire.

Fire can influence the microclimate of forest habitats by removing understory vegetation and surface debris. Temperature is often higher in recently burned forests owing to increased light penetration through the open understory. Because physiological processes are sensitive to temperature in ectotherms, we expected fire-maintained forests to improve the suitability of the thermal environment for turtles, and for turtles to seasonally associate with the most thermally-optimal habitats. Using a laboratory thermal gradient, we determined the thermal preference range (Tset) of eastern box turtles, Terrapene carolina, to be 27-31°C. Physical models simulating the body temperatures experienced by turtles in the field revealed that surface environments in a fire-maintained longleaf pine forest were 3°C warmer than adjacent unburned mixed hardwood/pine forests, but the fire-maintained forest was never of superior thermal quality owing to wider Te fluctuations above Tset and exposure to extreme and potentially lethal temperatures. Radiotracked turtles using fire-managed longleaf pine forests maintained shell temperatures (Ts) approximately 2°C above those at a nearby unburned forest, but we observed only moderate seasonal changes in habitat use which were inconsistent with thermoregulatory behavior. We conclude that turtles were not responding strongly to the thermal heterogeneity generated by fire in our system, and that other aspects of the environment are likely more important in shaping habitat associations.

Predicting bycatch hotspots for endangered leatherback turtles on longlines in the Pacific Ocean.

Fisheries bycatch is a critical source of mortality for rapidly declining populations of leatherback turtles, Dermochelys coriacea. We integrated use-intensity distributions for 135 satellite-tracked adult turtles with longline fishing effort to estimate predicted bycatch risk over space and time in the Pacific Ocean. Areas of predicted bycatch risk did not overlap for eastern and western Pacific nesting populations, warranting their consideration as distinct management units with respect to fisheries bycatch. For western Pacific nesting populations, we identified several areas of high risk in the north and central Pacific, but greatest risk was adjacent to primary nesting beaches in tropical seas of Indo-Pacific islands, largely confined to several exclusive economic zones under the jurisdiction of national authorities. For eastern Pacific nesting populations, we identified moderate risk associated with migrations to nesting beaches, but the greatest risk was in the South Pacific Gyre, a broad pelagic zone outside national waters where management is currently lacking and may prove difficult to implement. Efforts should focus on these predicted hotspots to develop more targeted management approaches to alleviate leatherback bycatch.

Temporal and spatial variation in landscape connectivity for a freshwater turtle in a temporally dynamic wetland system.

Inter-wetland connectivity, defined here as the movement of biota among discrete water bodies, can have important population- and community-level consequences in aquatic systems. We examined inter-wetland connectivity in a southeastern Australian national park by intensively monitoring the movements of freshwater turtles (Chelodina longicollis) via capture-mark-recapture over a three-year period, and more sporadically for 25 years. A high percentage (33%) of turtles moved between wetlands, suggesting that single wetlands should not represent the minimum habitat unit harboring a C. longicollis population. Distance was the only structural landscape metric correlated with inter-patch transition probability, with probability declining as inter-wetland distance increased. Movements also appear to be strongly influenced by shifting resource quality gradients between temporary wetlands and permanent lakes according to drought and flood cycles, a pattern more consistent with migration between critical resource patches than occasional interpopulational dispersal. Rare dispersal events of up to 5.2 km were known to occur. Captures at a terrestrial drift fence suggest that small and immature turtles moved between wetlands more frequently than our aquatic sampling indicated. We caution that measures of actual (or functional) connectivity can be biased by sampling methods and the temporal scale of sampling and must also be interpreted in the context of factors that motivate animal movements. This requires some understanding of spatial and temporal variation in intra-patch processes (e.g., quality and extent) and the expected movement responses of animals (e.g., habitat selection) over extended time frames, information that can potentially yield more important insight on connectivity than measures of landscape structural features alone.

Energy and water flux during terrestrial estivation and overland movement in a freshwater turtle.

The doubly labeled water (DLW) method for studying energy and water balance in field-active animals is not feasible for freshwater animals during aquatic activities, but several species of nominally aquatic reptiles leave wetlands for several critical and extended behaviors, where they face challenges to their energy and water balance. Using DLW, we studied energy and water relations during terrestrial estivation and movements in the eastern long-necked turtle (Chelodina longicollis), a species that inhabits temporary wetlands in southeastern Australia. Water efflux rates of 14.3-19.3 mL (kg d)(-1 ) during estivation were nearly offset by influx, indicating that turtles did not maintain water balance while terrestrial, though dehydration was slow. Estivation energy expenditure declined over time to 20.0-24.6 kJ (kg d)(-1) but did not indicate substantial physiological specializations. Energy reserves are predicted to limit survival in estivation to an estimated 49-261 d (depending on body fat), which is in close agreement with observed bouts of natural estivation in this population. The energy cost and water flux rates associated with overland movement behavior ranged from 46 to 99 kJ (kg d)(-1 ) and from 21.6 to 40.6 mL (kg d)(-1), respectively, for turtles moving 23-34 m d(-1). When a wetland dries, a turtle that forgoes movement to other wetlands can save sufficient energy to fuel up to 134 d in estivation. The increasing time in estivation with travel distance gained in this energy "trade-off" fits our previous observations that more turtles estivate when longer distances must be traveled to the nearest permanent lake, whereas emigration is nearly universal when only short distances must be traversed. The DLW method shows promise for addressing questions regarding the behavioral ecology and physiology of freshwater turtles in terrestrial situations, though validation studies are needed.

Maintenance of variable responses for coping with wetland drying in freshwater turtles.

Aquatic animals inhabiting temporary wetlands must respond to habitat drying either by estivating or moving to other wetlands. Using radiotelemetry and capture mark recapture, we examined factors influencing the decisions made by individuals in a population of freshwater turtles (Chelodina longicollis) in response to wetland drying in southeastern Australia. Turtles exhibited both behaviors, either remaining quiescent in terrestrial habitats for variable lengths of time (terrestrial estivation) or moving to other wetlands. Both the proportion of individuals that estivated terrestrially and the time individuals spent in terrestrial habitats increased with decreasing wetland hydroperiod and increasing distance to the nearest permanent wetland, suggesting behavioral decisions are conditional or state dependent (i.e., plastic) and influenced by local and landscape factors. Variation in the strategy or tactic chosen also increased with increasing isolation from other wetlands, suggesting that individuals differentially weigh the costs and benefits of residing terrestrially vs. those of long-distance movement; movement to other wetlands was the near universal strategy chosen when only a short distance must be traveled to permanent wetlands. The quality of temporary wetlands relative to permanent wetlands at our study site varies considerably and unpredictably with annual rainfall and with it the cost-benefit ratio of each strategy or tactic. Residency in or near temporary wetlands is more successful during wet periods due to production benefits, but movement to permanent wetlands is more successful, or least costly, during dry periods due to survival and body condition benefits. This shifting balance may maintain diversity in response of turtles to the spatial and temporal pattern in wetland quality if their response is in part genetically determined.

Effects of competition and coal-combustion wastes on recruitment and life history characteristics of salamanders in temporary wetlands.

Amphibians in natural systems must cope with a number of biotic and abiotic stressors that can potentially interact with pollutants to influence toxicity. Although interactive effects of short-lived pesticides with various environmental stressors have been studied, how persistent and bioaccumulative compounds such as metals interact with natural stressors to influence amphibians remains unexplored. We exposed the salamander Ambystoma talpoideum to coal-combustion wastes (a complex mixture of metals and metalloids, hereafter referred to as CCW) at low and high larval density throughout aquatic development in mesocosms simulating temporary wetlands. CCW and high density reduced survival to metamorphosis by 57-77% and 85-92%, respectively, and the effects of these two factors together were additive. Reduced metamorphosis was due in part to larval mortality prior to initiation of pond drying, but CCW and high density also extended the larval period, causing mortality of larvae that were not ready to metamorphose before the pond dried. To our knowledge, this is the first demonstration of a pollutant-induced extension of larval period leading to reduction in amphibian recruitment. Metamorphs were smaller in high density compared to low in reference ponds, but those from CCW emerged at similar sizes irrespective of density, suggesting less-than-additive effects of density and CCW on metamorph size. The adverse responses of salamanders to CCW were likely due to direct toxicity, as A. talpoideum metamorphs accumulated high concentrations of a suite of trace elements (As, Se, Sr, and V), and also to indirect effects on the community food web. We conclude that in no case did the addition of a natural stressor (high density) exacerbate CCW-related effects, but that the effects of CCW alone can be detrimental to larvae of salamanders that breed in temporary ponds.

Species- and stage-specific differences in trace element tissue concentrations in amphibians: implications for the disposal of coal-combustion wastes.

Information on species- and stage-specific patterns of contaminant accumulation is generally lacking for amphibians, yet such information could provide valuable knowledge on how amphibians interact with contaminants. We assessed concentrations of As, Cd, Cu, Ni, Pb, Se, Sr, and Zn in whole bodies of larval, recently metamorphosed, and adult life stages in Bufo terrestris and Rana sphenocephala from a site that currently receives coal combustion waste (CCW) discharge, a site where CCW was formerly discharged that has undergone natural attenuation for 30 years, and a nearby reference site. For the majority of elements (As, Cd, Cu, Ni, Pb, Zn), concentrations were highest in larvae, but Se and Sr concentrations remained elevated in later life stages, likely because these elements are S and Ca analogs, respectively, and are thus retained throughout structural changes during metamorphosis. Element concentrations were generally higher in B. terrestris than in R. sphenocephala. Concentrations of As, Se, and Sr were up to 11-35 times higher in metamorphs emigrating from CCW-polluted wetlands compared to unpolluted wetlands, suggesting metamorphosed amphibians can transport trace elements from aquatic disposal basins to nearby uncontaminated terrestrial habitats. In addition, anurans utilizing naturally revegetated sites up to 30 years after CCW disposal ceases are exposed to trace elements, although to a lesser degree than sites where CCW is currently discharged.

Functional relationships among selenium concentrations in the diet, target tissues, and nondestructive tissue samples of two species of snakes.

Nondestructive sampling methods, such as removal of feathers for contaminant analysis, are desirable in ecological monitoring programs that seek to minimize the impacts of harvesting organisms. Although many reptiles are declining worldwide, nondestructive sampling techniques seldom have been employed for assessing contaminant exposure in these organisms. In this study, we examined the utility of nondestructive tissue sampling for assessing Se exposure in reptiles. We describe the functional relationships among dietary Se concentrations, target tissue Se concentrations, and Se concentrations in nondestructive tissue samples (blood and tail tissue biopsy) in two species of snakes that had been exposed to Se under very different experimental protocols. Using nonlinear regression, we found strong positive correlations (r2 > 0.92) in all comparisons among Se concentrations in nondestructive tissues, diet, and target tissues. Moreover, equations describing these relationships can be used to estimate concentrations of Se in diet and target organs, from known concentrations of Se in nondestructive tissue samples. Although the current paucity of toxicity data on reptiles precludes tests of our models, we demonstrate how the equations describing these relationships might be used to make predictions about Se accumulation in target organs for risk assessment. Future studies on reptiles that examine these relationships under different Se exposure conditions, and those that document physiological responses of reptiles to various concentrations of Se, will help to refine our models and test their efficacy for predicting health risk.

Maternal transfer of selenium in Alligator mississippiensis nesting downstream from a coal-burning power plant.

Selenium (Se) is embryotoxic in many oviparous vertebrates, but little is known about maternal transfer of Se and its impact in reptiles. Over a four-year period, we collected three clutches of eggs of the American alligator (Alligator mississippiensis) from a single nest at a site contaminated with Se and compared egg and hatchling Se concentrations and clutch viability from this nest to nests downstream from the contaminated site (two clutches from two nests) and at a reference site (two clutches from two nests). Eggs and hatchlings from the nest at the Se-contaminated site and downstream nests had elevated Se concentrations (2.1-7.8 ppm) and lower viability (30-54%) compared to reference nests (1.4-2.3 ppm and 67-74% viability), but Se concentrations did not exceed reproductive toxicity thresholds established for other oviparous vertebrates. Selenium concentrations were higher in chorioallantoic membranes of eggs from Se-contaminated sites, suggesting that this tissue may be useful as a nondestructive index of Se exposure for embryos of A. mississippiensis. Examination of these data suggests that further studies on uptake, accumulation, and reproductive success of crocodilian embryos exposed to excessive Se are warranted.

Trophic and maternal transfer of selenium in brown house snakes (Lamprophis fuliginosus).

Excessive concentrations of dietary Se are toxic to oviparous vertebrates (i.e., fish and birds) but little is known about its accumulation and effects in reptiles. We exposed female brown house snakes, Lamprophis fuliginosus, to 10 and 20 microg/g Se by injecting seleno-D,L-methionine into their prey items and compared the snakes to individuals receiving background levels of approximately 1 microg/g dietary Se. Snakes were fed meals equaling 25% of their body mass 2-3 times a month for 10 months. Snakes exposed to excessive Se accumulated significant concentrations of Se in kidney, liver, and ovarian tissue, but accumulation had no effect on female survival, food consumption, growth, or body condition. Fewer females exposed to excessive Se reproduced than females exposed to 1 microg/g Se (67% vs. 91%, respectively), but the reduction in reproductive activity was not statistically significant. Total reproductive output of females did not differ among the three dietary treatments. However, snakes exposed to 10 and 20 microg/g Se transferred significant concentrations of Se to their eggs. In the 20 microg/g treatment, maternal transfer resulted in Se concentrations in eggs that surpassed all suggested reproductive toxicity thresholds for birds and fish. Further studies are needed to more rigorously determine whether maternal transfer of Se in this snake species affects the viability of developing embryos or the health of offspring.

Relationships among developmental stage, metamorphic timing, and concentrations of elements in bullfrogs (Rana catesbeiana).

We collected bullfrog (Rana catesbeiana) larvae from a coal combustion waste settling basin to investigate the effects of developmental stage and timing of metamorphosis on concentrations of a series of trace elements in bullfrog tissues. Bullfrogs at four stages of development (from no hind limbs to recently metamorphosed juveniles) and bullfrogs that metamorphosed in the fall or overwintered in the contaminated basin and metamorphosed in the spring were analyzed for whole-body concentrations of Al, V, Cr, Ni, Cu, As, Pb, Cd, Zn, Ag, Sr, and Se. After the effects of dry mass were removed, tissue concentrations of six elements (Al, V, Cr, Ni, Cu, As, and Pb) decreased from the late larval stage through metamorphosis. Decreases in concentrations through metamorphosis ranged from 40% for Cu to 97% for Al. Tissue concentrations of these elements were also similar or higher in spring; Al and Cr concentrations were 34 and 90% higher in the spring, respectively, whereas As, Ni, Cu, and Pb concentrations were <10% higher. Concentrations of Cd, Se, and Ag varied among seasons but not among stages; Cd and Ag concentrations were 40 and 62% lower, respectively, and Se concentrations were 21% higher in spring. Concentrations of Zn varied only among stages; concentrations decreased gradually through late larval stage and then increased through metamorphosis. Concentrations of Sr varied among stages, but this variation was dependent on the season. Concentrations of Sr were higher in larval stages during the spring, but because concentrations of Sr increased 122% through metamorphosis in the fall and only 22% in the spring, concentrations were higher in fall metamorphs when compared with spring metamorphs. Our results indicate that metamorphosis and season of metamorphosis affects trace element concentrations in bullfrogs and may have important implications for the health of juveniles and the transfer of pollutants from the aquatic to the terrestrial environment.

Effects of chronic dietary exposure to trace elements on banded water snakes (Nerodia fasciata).

Little currently is known about the accumulation or effects of contaminants on reptiles. To date, most studies examining reptile exposure to trace elements report tissue burdens of field-captured animals, but seldom provide insight into the dose, duration, or mode of exposure involved. For two years, we fed juvenile banded water snakes (Nerodia fasciata) prey items collected from a coal ash-contaminated site that contained elevated levels of As, Cd, Cu, Se, Sr, and V. With the exception of Cu, snakes accumulated significant concentrations of elements, usually in a dose-dependent manner. Accumulation varied significantly among liver, kidney, and gonads, and in most cases between sexes. Selenium accumulation was most notable, greatly exceeding established toxicity thresholds for other vertebrates. Despite the high concentrations of pollutants accumulated, snakes exposed to the contaminated diet survived through the study and exhibited normal food consumption, growth, condition factor, overwinter survival and mass loss, metabolic rate, and gonadosomatic index. The results of this study confirm that diet can be a significant route of exposure to trace elements in snakes and indicate that further studies on snakes are warranted to better understand their responses to contaminants.