Stream Salamanders are Relatively Tolerant of Salty Streams.

The application of road salt as a deicing agent is common, but investigations of potential negative effects of salt runoff on stream salamanders have been limited. Additionally, modern stormwater management practices should influence the delivery of salt ions to streams. We used data loggers in streams draining watersheds with and without stormwater management ponds and acute 96-h LC50 tests to investigate exposure of, and road salt toxicity among, two widespread salamanders in the Eastern USA, northern dusky salamanders (Desmognathus fuscus) and northern two-lined salamanders (Eurycea bislineata). In streams below stormwater ponds, base levels of conductivity were elevated throughout the year and elevated Cl- levels led to more frequent acute and chronic exceedances of U.S. Environmental Protection Agency ambient water quality criteria for Cl- when compared to streams draining watersheds with no stormwater management ponds. However, five of the six streams studied had exceedance frequencies suggesting Cl- associated with road salt application represented a persistent threat to aquatic life. Larval stream salamanders were relatively tolerant of salt, not exhibiting any lethal effects over a 96-h period until chloride levels exceeded 5000 mg/L for both species, and concentrations in streams rarely exceeded these levels and only for very short periods of time. Our results suggest road salts are not having acute lethal effects on salamanders in the streams we studied, but exceedance of U.S. Environmental Protection Agency ambient water quality standards for Cl- suggest the potential for sublethal and indirect effects of Cl- on salamander populations that require further study.

Influence of Modern Stormwater Management Practices on Transport of Road Salt to Surface Waters.

Application of road salts in regions with colder climates is leading to ground and surface water contamination. However, we know little about how modern stormwater management practices affect the movement of road salt through urban watersheds. We investigated groundwater contamination and transport of road salts at two stormwater ponds in Baltimore County, Maryland. In association with the ponds, we documented a plume of contaminated groundwater that resulted in Cl- loadings to the adjacent stream of 6574 to 40 008 kg Cl- per winter, depending on winter snowfall. We also monitored Na+ and Cl- ion concentrations and the temporal dynamics of conductivity at a range of stream sites in watersheds with and without stormwater management ponds. Streams draining watersheds with stormwater ponds had consistently higher conductivities and Cl- concentrations during base flow conditions and often exhibited greater peaks in Cl- and conductivity associated with winter storms and subsequent melting events, despite the degree of watershed development. Our results indicate that modern stormwater management practices are not protecting surface waters from road salt contamination and suggest they create contaminated plumes of groundwater that deliver Cl- and Na+ to streams throughout the year.

Precipitation influences on uptake of a global pollutant by a coastal avian species.

Climatic variation, including precipitation amounts and timing, has been linked to abundance and breeding success of many avian species. Less studied, but also of significance, is the consequence of climatic variability on the exposure and uptake of nutrients and contaminants by wildlife. The authors examined mercury (Hg) concentrations in nestling wood stork feathers in a coastal setting over a 16-yr period to understand the influence of rainfall amounts on Hg transfer by parental provisioning relative to habitat use, assuming differential bioavailability of Hg within freshwater and saltwater habitat types. Coastal Hg uptake by stork nestlings was linked to freshwater habitat use, as indicated by stable carbon isotope (δ(13)C) analyses. Cumulative rainfall amounts exceeding 220 cm in the 23 mo preceding the breeding seasons resulted in greater use of freshwater wetlands as foraging habitat and greater Hg accumulation by nestling storks.

Ameliorative effects of sodium chloride on acute copper toxicity among Cope's gray tree frog (Hyla chrysoscelis) and green frog (Rana clamitans) embryos.

Urban stormwater runoff is composed of a mixture of components, including polycyclic aromatic hydrocarbons, metals, deicing agents, and many others. The fate of these chemicals is often in stormwater detention ponds that are used by amphibians for breeding. Among aquatic organisms, the toxic mechanism for many metals involves interference with active Na(+) and Cl(-) uptake. Addition of cations has been shown to reduce the toxicity of metals among some aquatic organisms through competitive inhibition, but no studies have investigated the interaction between NaCl and Cu among amphibian embryos and larvae. To determine the degree to which NaCl may ameliorate the toxicity of Cu to amphibian embryos and larvae, the authors exposed Hyla chrysoscelis (Cope's gray treefrogs) and Rana (Lithobates) clamitans (green frogs) to seven levels of Cu and NaCl in fully factorial experiments. When exposure was in artificial hard water, Cu was highly toxic to both species (96-h median lethal concentration [LC50] of 44.7 µg/L and 162.6 µg/L for H. chrysoscelis and R. clamitans, respectively). However, approximately 500 mg/L of NaCl eliminated Cu toxicity over the range of Cu concentrations used in the experiments (maximum 150 µg Cu/L for H. chrysoscelis and 325 µg Cu/L for R. clamitans). The current results suggest that NaCl is likely responsible for the toxic effects of NaCl and metal mixtures that might be typical of runoff from road surfaces in northern latitudes.

An enriched stable isotope technique to estimate the availability of soil zinc to Lumbricus terrestris (L.) across a salinization gradient.

An enriched stable isotope approach was developed to evaluate Zn bioavailability to Lumbricus terrestris. The decrease in (68)Zn/(66) Zn in organ tissues was used to assess the relative magnitude of the bioavailable soil Zn pool. This tool was then used to specifically evaluate bioavailability as a function of soil cation distribution. Storm-water pond soils were modified using two treatment regimens whereby H(2)O-extractable Zn was varied either by different ZnCl(2) amendments or by constant ZnCl(2) amendment followed by varying the soil cation distribution through salt amendments (NaCl or CaCl(2)). Earthworms previously equilibrated in (68) Zn-spiked soil were introduced to experimental soils, and after 2 d, removed for analysis of isotopic ratios in specific tissues. Despite a wide range of H(2)O-extractable Zn values produced by the salt treatments (0.007-24.3 mg/kg), a significant relationship between Zn turnover rate in earthworm tissues and H(2)O-extractable Zn in the salt-treated soils was not observed. Rather, considering both treatment regimens, turnover rate better correlated with Zn present in broader pools, such as that extracted by 6M HNO(3). The bioavailability of trace metals to earthworms may be poorly characterized by loosely bound fractions such as the pore water. Additionally, the turnover rate of (68)Zn in anterior organ tissues may be an effective tool to evaluate the relative magnitude of the bioavailable soil Zn pool.

Effects of zinc exposure on earthworms, Lumbricus terrestris, in an artificial soil.

Earthworms have the potential to act as trophic links for pollutants that accumulate in urban soils. However, many pollutants may act as micronutrients at low concentrations and toxins at higher concentration. When pollutants are also micronutrients, bioaccumulations may initially increase trophic transfer as pollutant concentration increase, but at higher levels toxic effects may limit population size and the potential for trophic transfer. We found support for this model among earthworms exposed to a range of soil Zn levels. Worms showed increasing bioaccumulation of Zn with increasing Zn soil concentrations, but at higher Zn levels worm growth rates decreased.

Chemical fractionation of Cu and Zn in stormwater, roadway dust and stormwater pond sediments.

This study evaluated the chemical fractionation of Cu and Zn from source to deposition in a stormwater system. Cu and Zn concentrations and chemical fractionation were determined for roadway dust, roadway runoff and pond sediments. Stormwater Cu and Zn concentrations were used to generate cumulative frequency distributions to characterize potential exposure to pond-dwelling organisms. Dissolved stormwater Zn exceeded USEPA acute and chronic water quality criteria in approximately 20% of storm samples and 20% of the storm duration sampled. Dissolved Cu exceeded the previously published chronic criterion in 75% of storm samples and duration and exceeded the acute criterion in 45% of samples and duration. The majority of sediment Cu (92-98%) occurred in the most recalcitrant phase, suggesting low bioavailability; Zn was substantially more available (39-62% recalcitrant). Most sediment concentrations for Cu and Zn exceeded published threshold effect concentrations and Zn often exceeded probable effect concentrations in surface sediments.

Value of artificial habitats for amphibian reproduction in altered landscapes.

Installation and maintenance of stormwater ponds to detain and treat runoff from impervious surfaces is a common method of stormwater control in developed areas. That these ponds capture pollutants, however, is of concern for wildlife species that use the ponds, particularly pond-breeding amphibians. To assess the relative contribution of stormwater ponds to the persistence of amphibian populations in suburban landscapes, we compared amphibian use of stormwater ponds and other available wetlands in suburban and forested watersheds. We surveyed three suburban and three primarily forested first-order watersheds to identify all potential wetlands that might serve as breeding sites for pond-breeding amphibians. We performed call, egg-mass, and larval surveys to measure breeding effort at each wetland in spring and summer 2007 and 2008. In suburban watersheds most (89%) of the wetlands that had breeding activity were either stormwater ponds or otherwise artificial. This pattern was also evident in the forested watersheds, where amphibians were primarily found breeding in wetlands created by past human activity. Late-stage larvae were found only in anthropogenic wetlands in all study areas because the remaining natural wetlands did not hold water long enough for larvae to complete development. Our results suggest that in urban and suburban landscapes with naturally low densities of wetlands, wetlands created by current or historic land uses may be as important to amphibian conservation as natural wetlands or pools and that management strategies directed at urban and suburban landscapes should recognize and incorporate human-created wetlands.

Lethal and sublethal effects of embryonic and larval exposure of Hyla versicolor to Stormwater pond sediments.

Stormwater ponds are common features of modern stormwater management practices. Stormwater ponds often retain standing water for extended periods of time, develop vegetative characteristics similar to natural wetlands, and attract wildlife. However, because stormwater ponds are designed to capture pollutants, wildlife that utilize ponds might be exposed to pollutants and suffer toxicological effects. To investigate the toxicity of stormwater pond sediments to Hyla versicolor, an anuran commonly found using retention ponds for breeding, we exposed embryos and larvae to sediments in laboratory microcosms. Exposure to pond sediments reduced survival of embryos by approximately 50% but did not affect larval survival. Larvae exposed to stormwater pond sediment developed significantly faster (x = 39 days compared to 42 days; p = 0.005) and were significantly larger at metamorphosis (x = 0.49 g compared to 0.36 g; p < 0.001) than controls that were exposed to clean sand. Substantial amounts (712-2215 mg/l) of chloride leached from pond sediments into the water column of treatment microcosms; subsequently, survival of embryos was negatively correlated (r (2) = 0.50; p < 0.001) with water conductivity during development. Our results, along with the limited number of other toxicological studies of stormwater ponds, suggest that road salt contributes to the degradation of stormwater pond habitat quality for amphibian reproduction and that future research should focus on understanding interactions among road salts and other pollutants and stressors characteristic of urban environments.

Impacts of weathered tire debris on the development of Rana sylvatica larvae.

Highway runoff has the potential to negatively impact receiving systems including stormwater retention ponds where highway particulate matter can accumulate following runoff events. Tire wear particles, which contain about 1% Zn by mass, make up approximately one-third of the vehicle derived particulates in highway runoff and therefore may serve as a stressor to organisms utilizing retention ponds as habitat. In this study, we focused on the potential contribution of tire debris to Zn accumulation by Rana sylvatica larvae and possible lethal or sublethal impacts resulting from exposure to weathered tire debris during development. Eggs and larvae were exposed to aged sediments (containing either ZnCl2 or tire particulate matter, both providing nominal concentrations of 1000 mg Zn kg(-1)) through metamorphosis. Water column Zn was elevated in both the ZnCl2 and tire treatments relative to the control treatment, indicating that aging allowed Zn leaching from tire debris to occur. Tissue Zn was also elevated for the ZnCl2 and tire treatments indicating that Zn in the treatments was available for uptake by the amphibians. Exposure to both ZnCl2 and tire treatments increased the time for larvae to complete metamorphosis in comparison with controls. We also observed that the longer the organisms took to complete metamorphosis, the smaller their mass at metamorphosis. Our results indicate that Zn leached from aged tire debris is bioavailable to developing R. sylvatica larvae and that exposure to tire debris amended sediments can result in measurable physiological outcomes to wood frogs that may influence population dynamics.

Environmental correlates, plasticity, and repeatability of differences in performance among blacknose dace (Rhinichthys atratulus) populations across a gradient of urbanization.

Urbanization alters stream and watershed hydrology so that fish from urban stream systems are confronted with extreme flows during storms and runoff events. To test whether residence in urban streams is associated with altered swimming ability, we compared sprint and endurance swimming performances of eight populations of blacknose dace (Rhinichthys atratulus) from different watersheds along an urban/rural gradient. Watershed impervious surface cover, a measure of urbanization, was significantly correlated with sprint performance in dace from all stream types and endurance swimming performance (U(crit)) when only fish from urban streams were analyzed. Three estimators of water flow in a stream system, watershed area, mean annual discharge, and base-flow current speed, were all related to U(crit) in fish from nonurban streams. The U(crit) was significantly repeatable after 6 mo in the laboratory, but dace populations with exceptional U(crit) values lost ability under no-flow, "detraining" conditions. Sprint performance changed substantially in individual dace after 10 wk under no-flow conditions and was a significant function of the animal's original performance. Animals with high sprint performance tended to lose ability, whereas those with poor performance gained ability. Interpopulation differences in both sprint and endurance swimming were robust over multiple years of collection from the same sites.

Microcosm investigations of stormwater pond sediment toxicity to embryonic and larval amphibians: variation in sensitivity among species.

Stormwater ponds have become common features of modern development and often represent significant amounts of open space in urbanized areas. Although stormwater ponds may provide habitat for wildlife, factors responsible for producing variation in wildlife use of ponds have received limited attention. To investigate the role of variation in species tolerances of pollutants in structuring pond-breeding amphibian assemblages, we exposed species tolerant (Bufo americanus) and not tolerant (Rana sylvatica) of urbanization to pond sediments in laboratory microcosms. Pond microcosms had elevated sediment metal levels and chloride water concentrations. Among R. sylvatica embryos, exposure to pond sediments resulted in 100% mortality. In contrast, B. americanus embryos and larvae experienced only sublethal effects (i.e., reduced size at metamorphosis) due to pond sediment exposure. Our results suggest variation in pollutant tolerance among early developmental stages of amphibians may act in concert with terrestrial habitat availability to structure amphibian assemblages associated with stormwater ponds.

Climatic variation and the distribution of an amphibian polyploid complex.

The establishment of polyploid populations involves the persistence and growth of the polyploid in the presence of the progenitor species. Although there have been a number of animal polyploid species documented, relatively few inquiries have been made into the large-scale mechanisms of polyploid establishment in animal groups. Herein we investigate the influence of regional climatic conditions on the distributional patterns of a diploid-tetraploid species pair of gray treefrogs, Hyla chrysoscelis and H. versicolor (Anura: Hylidae) in the mid-Atlantic region of eastern North America. Calling surveys at breeding sites were used to document the distribution of each species. Twelve climatic models and one elevation model were generated to predict climatic and elevation values for gray treefrog breeding sites. A canonical analysis of discriminants was used to describe relationships between climatic variables, elevation and the distribution of H. chrysoscelis and H. versicolor. There was a strong correlation between several climatic variables, elevation and the distribution of the gray treefrog complex. Specifically, the tetraploid species almost exclusively occupied areas of higher elevation, where climatic conditions were relatively severe (colder, drier, greater annual variation). In contrast, the diploid species was restricted to lower elevations, where climatic conditions were warmer, wetter and exhibited less annual variation. Clusters of syntopic sites were associated with areas of high variation in annual temperature and precipitation during the breeding season. Our data suggest that large-scale climatic conditions have played a role in the establishment of the polyploid H. versicolor in at least some portions of its range. The occurrence of the polyploid and absence of the progenitor in colder, drier and more varied environments suggests the polyploid may possess a tolerance of severe environmental conditions that is not possessed by the diploid progenitor. Our findings support the hypothesis that increased tolerance to severe environmental conditions is a plausible mechanism of polyploid establishment.

Influence of larval period on responses of overwintering green frog (Rana clamitans) larvae exposed to contaminated sediments.

Pond-breeding amphibians exhibit large intra- and interspecific differences in the duration of the aquatic larval phase. In contaminated aquatic environments, a prolonged larval phase means prolonged exposure to pollutants and, potentially, more severe toxic effects. In the laboratory, we tested this hypothesis by exposing green frog larvae (Rana clamitans) to commercial clean sand (control), sediment from an abandoned surface mine (mine), or sediment contaminated with coal combustion waste (CCW). By collecting eggs late in the breeding season, we obligated larvae to overwinter and spend a protracted amount of time exposed to contaminated sediments. The experiment was continued until all larvae either successfully completed metamorphosis or died (301 d). Larvae exposed to mine sediments accumulated significant levels of Pb and Zn, whereas larvae exposed to CCW-contaminated sediment accumulated significant levels of As, Se, Sr, and V. Larvae exposed to mine sediments suffered sublethal effects in the form of reduced growth and size at metamorphosis, but the proportion of larvae successfully completing metamorphosis (93%) was the same for both control and mine treatments. In contrast, larvae exposed to CCW-contaminated sediment suffered greatly reduced survival (13%) compared to both control and mine treatments. Moreover, among larvae in the CCW treatment, the majority of mortality occurred during the latter part the overwintering period (after day 205), corresponding to the onset of metamorphosis in the controls. Our results suggest that the length of the larval period may be one of many life-history or ecological characteristics that influence the sensitivity of aquatic breeding amphibians to environmental pollutants.

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.

Species-specific responses of developing anurans to coal combustion wastes.

Field surveys and field experiments have previously documented adverse effects of solid byproducts from coal incineration (coal combustion wastes (CCW)) on larval amphibians inhabiting aquatic habitats. However, a definitive link between CCW-exposure and developmental abnormalities has not been established because no studies have addressed the direct effects of prolonged exposure to CCW on larval amphibian development under controlled laboratory conditions. In the laboratory we exposed green frog (Rana clamitans) and wood frog (Rana sylvatica) larvae to either clean sand or CCW-contaminated sediment to investigate the direct effects of CCW exposure on trace element accumulation, growth, developmental rate, malformations, survival, and metamorphic success. While both species accumulated significant (P < 0.05) concentrations of at least six trace elements (As, Cd, Fe, Se, Sr, and V), effects of exposure to CCW varied between species, with R. clamitans larvae experiencing more severe effects including a 26% reduction in survival and a 45% reduction in metamorphic success. Furthermore, exposure to CCW decreased growth and developmental rates among larvae of both species that successfully completed metamorphosis. Larval period duration was increased by 10 and 11%, and size at metamorphosis was decreased by 10 and 39% in R. clamitans and R. sylvatica exposed to CCW, respectively. Rates of malformations were 50 million t are discharged annually to surface impoundments in the US, which are often used by breeding amphibians.

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.