Aaron B. Stoler

Leaf litter quality induces morphological and developmental changes in larval amphibians.

Aquatic consumers exhibit many types of inducible phenotypic responses to variation in resource quantity and quality. Leaf litter constitutes a primary resource in freshwater systems, and variation in litter quality can alter the growth and development of aquatic consumers. It is therefore reasonable to hypothesize that variation in litter quality might also induce phenotypic changes in consumers. To test this hypothesis, we exposed two densities of wood frog (Lithobates sylvaticus [Rana sylvatica]) tadpoles to six chemically distinct species of leaf litter from temperate broadleaf and coniferous trees. After several weeks, we quantified development rate, growth rate, intestinal length, size of the oral disc, and five external dimensions of the tadpoles. In addition to substantial changes in growth and development rates, we found striking changes in all morphological responses among different leaf litter environments, including up to 14% longer intestines, 11% deeper tails, and 6% deeper tail muscles. In addition, we found strong relationships of total nitrogen content with all morphological features except growth rate. Our results indicate that differences in resource quality can induce phenotypic changes that are as large as or larger than changes induced by resource quantity. Our study also has substantial implications for the future of aquatic consumers living in forested wetlands given that these forests are currently experiencing widespread changes in tree composition.

Cross-tolerance in amphibians: wood frog mortality when exposed to three insecticides with a common mode of action.

Insecticide tolerance and cross-tolerance in nontarget organisms is often overlooked despite its potential to buffer natural systems from anthropogenic influence. We exposed wood frog tadpoles from 15 populations to three acetylcholine esterase-inhibiting insecticides and found widespread variation in insecticide tolerance and evidence for cross-tolerance to these insecticides. Our results demonstrate that amphibian populations with tolerance to one pesticide may be tolerant to many other pesticides.

Bottom-up meets top-down: leaf litter inputs influence predator–prey interactions in wetlands

While the common conceptual role of resource subsidies is one of bottom-up nutrient and energy supply, inputs can also alter the structural complexity of environments. This can further impact resource flow by providing refuge for prey and decreasing predation rates. However, the direct influence of different organic subsidies on predator–prey dynamics is rarely examined. In forested wetlands, leaf litter inputs are a dominant energy and nutrient resource and they can also increase benthic surface cover and decrease water clarity, which may provide refugia for prey and subsequently reduce predation rates. In outdoor mesocosms, we investigated how inputs of leaf litter that alter benthic surface cover and water clarity influence the mortality and growth of gray treefrog tadpoles (Hyla versicolor) in the presence of free-swimming adult newts (Notophthalmus viridiscens), which are visual predators. To manipulate surface cover, we added either oak (Quercus spp.) or red pine (Pinus resinosa) litter and crossed these treatments with three levels of red maple (Acer rubrum) litter leachate to manipulate water clarity. In contrast to our predictions, benthic surface cover had no effect on tadpole survival while darkening the water caused lower survival. In addition, individual tadpole mass was lowest in the high maple leachate treatments, suggesting an interaction between bottom-up effects of leaf litter and top-down effects of predation risk that altered mortality and growth of tadpoles. Our results indicate that realistic changes in forest tree composition, which cause concomitant changes in litter inputs to wetlands, can substantially alter community interactions.

Effects of Pesticide Mixtures on Host-Pathogen Dynamics of the Amphibian Chytrid Fungus.

Anthropogenic and natural stressors often interact to affect organisms. Amphibian populations are undergoing unprecedented declines and extinctions with pesticides and emerging infectious diseases implicated as causal factors. Although these factors often co-occur, their effects on amphibians are usually examined in isolation. We hypothesized that exposure of larval and metamorphic amphibians to ecologically relevant concentrations of pesticide mixtures would increase their post-metamorphic susceptibility to the fungus Batrachochytrium dendrobatidis (Bd), a pathogen that has contributed to amphibian population declines worldwide. We exposed five anuran species (Pacific treefrog, Pseudacris regilla; spring peeper, Pseudacris crucifer; Cascades frog, Rana cascadae; northern leopard frog, Lithobates pipiens; and western toad, Anaxyrus boreas) from three families to mixtures of four common insecticides (chlorpyrifos, carbaryl, permethrin, and endosulfan) or herbicides (glyphosate, acetochlor, atrazine, and 2,4-D) or a control treatment, either as tadpoles or as newly metamorphic individuals (metamorphs). Subsequently, we exposed animals to Bd or a control inoculate after metamorphosis and compared survival and Bd load. Bd exposure significantly increased mortality in Pacific treefrogs, spring peepers, and western toads, but not in Cascades frogs or northern leopard frogs. However, the effects of pesticide exposure on mortality were negligible, regardless of the timing of exposure. Bd load varied considerably across species; Pacific treefrogs, spring peepers, and western toads had the highest loads, whereas Cascades frogs and northern leopard frogs had the lowest loads. The influence of pesticide exposure on Bd load depended on the amphibian species, timing of pesticide exposure, and the particular pesticide treatment. Our results suggest that exposure to realistic pesticide concentrations has minimal effects on Bd-induced mortality, but can alter Bd load. This result could have broad implications for risk assessment of amphibians; the outcome of exposure to multiple stressors may be unpredictable and can differ between species and life stages.

Differential consumption and assimilation of leaf litter by wetland herbivores: alternative pathways for decomposition and trophic transfer

The process of decomposition has received much attention in terrestrial and stream ecosystems, but our understanding of the factors that contribute to this process in wetlands remains relatively poor. Many macroconsumers in wetlands are classically labeled as herbivores, but increasing evidence suggests that they also contribute to the breakdown of dead plant litter depending on the nutritional quality (i.e., nutrient content, density, and toxicity) of the litter. We examined the relative contributions of 2 common North American temperate wetland consumers, the green frog tadpole (Lithobates clamitans) and the Ram’s Horn snail (Planorbella trivolvis), to the decomposition of 5 chemically variable plant litter species. Based on anatomical differences (e.g., mouth parts, digestive structures), we hypothesized that snails would have higher consumption rates than tadpoles, but that tadpoles would have higher assimilation efficiency. We also predicted that consumption rates and assimilation efficiency would vary with litter nutritional quality. Overall, consumers exhibited substantial detritivory and caused up to 62% litter mass loss relative to treatments with only microbes. As hypothesized, snails consumed more than tadpoles, but this difference was largely explained by differences in consumer mass. Contrary to our hypothesis, snails and tadpoles exhibited similar assimilation efficiencies. Both litter mass loss and assimilation efficiency by consumers differed among litter species treatments. Litter mass loss tended to be negatively correlated with litter C∶N and C∶P, whereas assimilation efficiency had no detectable correlation with any measured litter characteristic. Our study demonstrates that studies of energy and nutrient budgets in wetlands should consider both consumer type and litter species to describe ecosystem function fully.

Leaves and litterbugs: how litter quality affects amphipod life-history and sexually selected traits

Abstract: Resources that cross ecosystem boundaries (subsidies) are common in nature, but we have little knowledge of how such resources affect individual traits and, consequently, ecological interactions. In wetlands, leaf litter is an important resource subsidy that regulates ecological processes including the production of consumer biomass. However, litter quality is highly variable and depends on local plant species diversity. Many aquatic consumers are highly nutrient limited, so differences in nutrients and other chemical components in litter are likely to affect numerous individual traits of consumers including life-history and sexually selected traits. We tested whether the species of tree leaf litter consumed by freshwater amphipods affected survival, growth, and development of sexually selected traits. Amphipods had higher survival, attained larger body sizes, and had larger male sexual traits when reared on litter that was relatively nutritious and low in plant secondary and structural compounds. Such widespread trait changes suggest that changing forest composition could have substantial effects on wetland ecology and the evolution of sexual traits over ecological time scales.

Combined effects of road salt and an insecticide on wetland communities

As the numbers of chemical contaminants in freshwater ecosystems increase, it is important to understand whether contaminants interact in ecologically important ways. The present study investigated the independent and interactive effects of 2 contaminants that frequently co-occur in freshwater environments among higher latitudes, including a commonly applied insecticide (carbaryl) and road salt (NaCl). The hypothesis was that the addition of either contaminant would result in a decline in zooplankton, an algal bloom, and the subsequent decline of both periphyton and periphyton consumers. Another hypothesis was that combining the contaminants would result in synergistic effects on community responses. Outdoor mesocosms were used with communities that included phytoplankton, periphyton, zooplankton, amphipods, clams, snails, and tadpoles. Communities were exposed to 4 environmentally relevant concentrations of salt (27 mg Cl- L-1 , 77 mg Cl- L-1 , 277 mg Cl- L-1 , and 727 mg Cl- L-1 ) fully crossed with 4 carbaryl treatments (ethanol, 0 µg L-1 , 5 µg L-1 , and 50 µg L-1 ) over 57 d. Contaminants induced declines in rotifer and cladoceran zooplankton, but only carbaryl induced an algal bloom. Consumers exhibited both positive and negative responses to contaminants, which were likely the result of both indirect community interactions and direct toxicity. In contrast to the hypothesis, no synergistic effects were found, although copepod densities declined when high concentrations of both chemicals were combined. The results suggest that low concentrations of salt and carbaryl are likely to have mostly independent effects on aquatic communities. Environ Toxicol Chem 2017;36:771-779.

Leaf Litter Inhibits Growth of an Amphibian Fungal Pathogen

Past studies have found a heterogeneous distribution of the amphibian chytrid fungal pathogen, Batrachochytrium dendrobatidis (Bd). Recent studies have accounted for some of this heterogeneity through a positive association between canopy cover and Bd abundance, which is attributed to the cooling effect of canopy cover. We questioned whether leaf litter inputs that are also associated with canopy cover might also alter Bd growth. Leaf litter inputs exhibit tremendous interspecific chemical variation, and we hypothesized that Bd growth varies with leachate chemistry. We also hypothesized that Bd uses leaf litter as a growth substrate. To test these hypotheses, we conducted laboratory trials in which we exposed cultures of Bd to leachate of 12 temperate leaf litter species at varying dilutions. Using a subset of those 12 litter species, we also exposed Bd to pre-leached litter substrate. We found that exposure to litter leachate and substrate reduced Bd spore and sporangia densities, although there was substantial variation among treatments. In particular, Bd densities were inversely correlated with concentrations of phenolic acids. We conducted a field survey of phenolic concentrations in natural wetlands which verified that the leachate concentrations in our lab study are ecologically relevant. Our study reinforces prior indications that positive associations between canopy cover and Bd abundance are likely mediated by water temperature effects, but this phenomenon might be counteracted by changes in aquatic chemistry from leaf litter inputs.

Ontogenetic changes in sensitivity to nutrient limitation of tadpole growth

According to ecological stoichiometry (ES), the growth of a consumer with abundant resources should increase as body and resource stoichiometry become more similar. However, for organisms with complex life cycles involving distinct changes in biology, nutrient demands might change in response to ontogenetic changes in body stoichiometry. Tadpole growth and development has been found to be largely nitrogen (N) limited, as predicted for organisms developing N-rich tissues like muscle. However, tadpole metamorphosis includes periods of rapid development of phosphorus (P)-rich bones in preparation for a terrestrial lifestyle. We hypothesized that tadpole growth and development will exhibit variable nutrient demands during different stages of ontogeny, due to predictable changes in body tissue stoichiometry. To test this, we raised tadpoles on four diets with varying N:P ratios and assessed growth and developmental rates. Specifically, we predicted that tadpoles would be sensitive to N limitation throughout ontogeny (consistent with previous studies), but also sensitive to P limitation during the process of long-bone ossification. Consistent with our prediction, tadpole growth rates and development were sensitive to N limitation throughout ontogeny. Increased dietary N led to a shorter time to metamorphosis and a larger mass at metamorphosis. Also as predicted, growth rates were sensitive to both N and P during the period of peak bone ossification, indicative of co-limitation. These results indicate that P limitation changes through tadpole ontogeny consistent with, and can be predicted by, shifts in body tissue stoichiometry. Future studies should investigate whether ontogenetic shifts in tadpole P limitation lead to seasonal shifts in wetland nutrient cycling.

Leaf litter mediates the negative effect of road salt on forested wetland communities

Human modification of landscapes has substantially altered the quality and quantity of terrestrial subsidies to freshwater ecosystems. The same modifications frequently lead to addition of chemical contaminants to freshwater environments. Both types of environmental change can alter the abundance of species and can lead to ecological interactions that affect entire communities. We examined how variation of tree litter inputs interacts with inputs of road salt deicers, which are an increasingly common contaminant in northern latitudes. Based on studies of the effects of each factor in isolation, we hypothesized that elevated Cl− levels would reduce copepod densities, increase algal abundance, and subsequently increase salt-tolerant consumer densities and biomass. We also hypothesized that these effects would be most pronounced in the presence of highly soluble leaf litter (e.g., Acer rubrum). We constructed experimental freshwater ponds containing assemblages of phytoplankton, periphyton, zooplankton, Physa acuta snails, and 2 species of tadpoles (Lithobates sylvaticus and Anaxyrus americanus). We used a fully factorial design, manipulating leaf litter (none, A. rubrum, or Quercus velutina) and Cl− concentration (114, 220, 314, and 867 mg Cl/L). Road salt at the 3 lower concentrations had few significant effects. The highest Cl− concentration reduced copepod densities and increased phytoplankton concentrations, but only in the presence of maple litter. We also observed increased rotifer densities in the highest Cl− concentration, but only in the presence of either litter species. Our results indicate that road salt contamination can have significant effects on wetland community composition at relatively high concentrations, but these effects depend on the chemistry of allochthonous inputs.