Assessing Leukocyte Profiles of Salamanders and Other Amphibians: A Herpetologists' Guide.

Assessing numbers of leukocytes in salamanders and other amphibians can be useful metrics for understanding health or stress levels of individuals in a population. In this chapter we describe the procedures for obtaining blood samples from amphibians, preparing blood films for microscopy, counting, and identifying cells. We also provide reference values for amphibian leukocytes for use in interpreting leukocyte data. From our assessment of the published and unpublished literature, "non-stressed" salamanders would have a leukocyte profile where 60-70% of cells are lymphocytes, 17-30% are neutrophils, 1-4% are eosinophils, 4-12% are basophils, and 2-6% are monocytes. In Ambystoma spp., the eosinophil abundance can be notably higher (30% of all white blood cells), for reasons unknown. Finally, the neutrophil-lymphocyte ratio of most non-stressed salamanders tends to be between 0.3 and 0.4 (sometimes less), while the ratios of stressed salamanders tend to be over 1.0.

Sex-related differences in aging rate are associated with sex chromosome system in amphibians.

Sex-related differences in mortality are widespread in the animal kingdom. Although studies have shown that sex determination systems might drive lifespan evolution, sex chromosome influence on aging rates have not been investigated so far, likely due to an apparent lack of demographic data from clades including both XY (with heterogametic males) and ZW (heterogametic females) systems. Taking advantage of a unique collection of capture-recapture datasets in amphibians, a vertebrate group where XY and ZW systems have repeatedly evolved over the past 200 million years, we examined whether sex heterogamy can predict sex differences in aging rates and lifespans. We showed that the strength and direction of sex differences in aging rates (and not lifespan) differ between XY and ZW systems. Sex-specific variation in aging rates was moderate within each system, but aging rates tended to be consistently higher in the heterogametic sex. This led to small but detectable effects of sex chromosome system on sex differences in aging rates in our models. Although preliminary, our results suggest that exposed recessive deleterious mutations on the X/Z chromosome (the "unguarded X/Z effect") or repeat-rich Y/W chromosome (the "toxic Y/W effect") could accelerate aging in the heterogametic sex in some vertebrate clades.

Effects of Salinity on Hatchling Diamond-Backed Terrapin (Malaclemys terrapin) Growth, Behavior, and Stress Physiology.

Diamond-backed Terrapins inhabit coastal salt marshes along the eastern and Gulf coasts of North America. Terrapins are adapted to intermediate salinities yet frequently face saltwater-inundated marsh habitat exceeding 25 ppt (or grams/kilogram). We investigated the effect of salinity on the growth of hatchling terrapins and on their compensatory responses to salinity stress. We randomly assigned 30 terrapin hatchlings each to one of five salinity treatments (1, 5, 10, 20, or 35 ppt). Over 75 d, we regularly monitored behavior, appetite, and changes in growth; and calculated ratios of heterophils to lymphocytes (H:L ratio) to assess responses to prolonged salinity stress. Consistent with prior studies, chronic exposure to high salinity significantly reduced hatchling growth. Hatchlings in 20-ppt and 35-ppt salinities exhibited appetite suppression and saltwater avoidance and were more likely to show freshwater-seeking behaviors. H:L ratios were higher among hatchlings in 20-and 35-ppt salinities, consistent with a corticosterone-driven stress response to sustained high-salinity exposure, which may play a role in limiting growth. Our findings suggest hatchling growth and distribution among local habitats will vary spatially depending on habitat salinity and freshwater accessibility. The growth-limiting effects of chronically high salinity or limited access to freshwater could therefore increase hatchling mortality and be an important driver of spatial variation in terrapin demography and abundance. However, when freshwater sources are available, compensatory behaviors might reduce growth-limiting effects. Terrapin recruitment is likely to be impacted as rising sea levels, increased human water use, land development, and other anthropogenic changes alter freshwater inputs to coastal marshes.

Predicted alteration of surface activity as a consequence of climate change.

Wildlife are faced with numerous threats to survival, none more pressing than that of climate change. Understanding how species will respond behaviorally, physiologically, and demographically to a changing climate is a cornerstone of many contemporary ecological studies, especially for organisms, such as amphibians, whose persistence is closely tied to abiotic conditions. Activity is a useful parameter for understanding the effects of climate change because activity is directly linked to fitness as it dictates foraging times, energy budgets, and mating opportunities. However, activity can be challenging to measure directly, especially for secretive organisms like plethodontid salamanders, which only become surface active when conditions are cool and moist because of their anatomical and physiological restrictions. We estimated abiotic predictors of surface activity for the seven species of the Plethodon jordani complex. Five independent data sets collected from 2004 to 2017 were used to determine the parameters driving salamander surface activity in the present day, which were then used to predict potential activity changes over the next 80 yrs. Average active seasonal temperature and vapor pressure deficit were the strongest predictors of salamander surface activity and, without physiological or behavioral modifications, salamanders were predicted to exhibit a higher probability of surface activity during peak active season under future climate conditions. Temperatures during the active season likely do not exceed salamander thermal maxima to cause activity suppression and, until physiological limits are reached, future conditions may continue to increase activity. Our model is the first comprehensive field-based study to assess current and future surface activity probability. Our study provides insights into how a key behavior driving fitness may be affected by climate change.

Experimental N and P additions relieve stoichiometric constraints on organic matter flows through five stream food webs.

Human activities have dramatically altered global patterns of nitrogen (N) and phosphorus (P) availability. This pervasive nutrient pollution is changing basal resource quality in food webs, thereby affecting rates of biological productivity and the pathways of energy and material flow to higher trophic levels. Here, we investigate how the stoichiometric quality of basal resources modulates patterns of material flow through food webs by characterizing the effects of experimental N and P enrichment on the trophic basis of macroinvertebrate production and flows of dominant food resources to consumers in five detritus-based stream food webs. After a pre-treatment year, each stream received N and P at different concentrations for 2 years, resulting in a unique dissolved N:P ratio (target range from 128:1 to 2:1) for each stream. We combined estimates of secondary production and gut contents analysis to calculate rates of material flow from basal resources to macroinvertebrate consumers in all five streams, during all 3 years of study. Nutrient enrichment resulted in a 1.5× increase in basal resource flows to primary consumers, with the greatest increases from biofilms and wood. Flows of most basal resources were negatively related to resource C:P, indicating widespread P limitation in these detritus-based food webs. Nutrient enrichment resulted in a greater proportion of leaf litter, the dominant resource flow-pathway, being consumed by macroinvertebrates, with that proportion increasing with decreasing leaf litter C:P. However, the increase in efficiency with which basal resources were channelled into metazoan food webs was not propagated to macroinvertebrate predators, as flows of prey did not systematically increase following enrichment and were unrelated to basal resource flows. This study suggests that ongoing global increases in N and P supply will increase organic matter flows to metazoan food webs in detritus-based ecosystems by reducing stoichiometric constraints at basal trophic levels. However, the extent to which those flows are propagated to the highest trophic levels likely depends on responses of individual prey taxa and their relative susceptibility to predation.

Side-swiped: Ecological cascades emanating from earthworm invasion.

Non-native, invasive earthworms are altering soils throughout the world. Ecological cascades emanating from these changes stem from earthworm-caused changes in detritus processing occurring at a mid-point in the trophic pyramid, rather than the more familiar bottom-up or top-down cascades. They include fundamental changes (microcascades) in soil morphology, bulk density, nutrient leaching, and a shift to warmer, drier soil surfaces with loss of organic horizons. In North American temperate and boreal forests, microcascades cause effects of concern to society (macrocascades), including changes in CO2 sequestration, disturbance regimes, soil quality, water quality, forest productivity, plant communities, and wildlife habitat, and facilitation of other invasive species. Interactions among these changes create cascade complexes that interact with climate change and other environmental changes. The diversity of cascade effects, combined with the vast area invaded by earthworms, lead to regionally important changes in ecological functioning.

Migratory monarchs that encounter resident monarchs show life-history differences and higher rates of parasite infection.

Environmental change induces some wildlife populations to shift from migratory to resident behaviours. Newly formed resident populations could influence the health and behaviour of remaining migrants. We investigated migrant-resident interactions among monarch butterflies and consequences for life history and parasitism. Eastern North American monarchs migrate annually to Mexico, but some now breed year-round on exotic milkweed in the southern US and experience high infection prevalence of protozoan parasites. Using stable isotopes (δ2 H, δ13 C) and cardenolide profiles to estimate natal origins, we show that migrant and resident monarchs overlap during fall and spring migration. Migrants at sites with residents were 13 times more likely to have infections and three times more likely to be reproductive (outside normal breeding season) compared to other migrants. Exotic milkweed might either attract migrants that are already infected or reproductive, or alternatively, induce these states. Increased migrant-resident interactions could affect monarch parasitism, migratory success and long-term conservation.

Litter P content drives consumer production in detritus-based streams spanning an experimental N:P gradient.

Ecological stoichiometry theory (EST) is a key framework for predicting how variation in N:P supply ratios influences biological processes, at molecular to ecosystem scales, by altering the availability of C, N, and P relative to organismal requirements. We tested EST predictions by fertilizing five forest streams at different dissolved molar N:P ratios (2, 8, 16, 32, 128) for two years and tracking responses of macroinvertebrate consumers to the resulting steep experimental gradient in basal resource stoichiometry (leaf litter %N, %P, and N:P). Nitrogen and P content of leaf litter, the dominant basal resource, increased in all five streams following enrichment, with steepest responses in litter %P and N:P ratio. Additionally, increases in primary consumer biomass and production occurred in all five streams following N and P enrichment (averages across all streams: biomass by 1.2×, production by 1.6×). Patterns of both biomass and production were best predicted by leaf litter N:P and %P and were unrelated to leaf litter %N. Primary consumer production increased most in streams where decreases in leaf litter N:P were largest. Macroinvertebrate predator biomass and production were also strongly positively related to litter %P, providing robust experimental evidence for the primacy of P limitation at multiple trophic levels in these ecosystems. However, production of predatory macroinvertebrates was not related directly to primary consumer production, suggesting the importance of additional controls for macroinvertebrates at upper trophic positions. Our results reveal potential drivers of animal production in detritus-based ecosystems, including the relative importance of resource quality vs. quantity. Our study also sheds light on the more general impacts of variation in N:P supply ratio on nutrient-poor ecosystems, providing strong empirical support for predictions that nutrient enrichment increases food web productivity whenever large elemental imbalances between basal resources and consumer demand are reduced.

Habitat predictors of genetic diversity for two sympatric wetland-breeding amphibian species.

Population genetic diversity is widely accepted as important to the conservation and management of wildlife. However, habitat features may differentially affect evolutionary processes that facilitate population genetic diversity among sympatric species. We measured genetic diversity for two pond-breeding amphibian species (Dwarf salamanders, Eurycea quadridigitata; and Southern Leopard frogs, Lithobates sphenocephalus) to understand how habitat characteristics and spatial scale affect genetic diversity across a landscape. Samples were collected from wetlands on a longleaf pine reserve in Georgia. We genotyped microsatellite loci for both species to assess population structures and determine which habitat features were most closely associated with observed heterozygosity and rarefied allelic richness. Both species exhibited significant population genetic structure; however, structure in Southern Leopard frogs was driven primarily by one outlier site. Dwarf salamander allelic richness was greater at sites with less surrounding road area within 0.5 km and more wetland area within 1.0 and 2.5 km, and heterozygosity was greater at sites with more wetland area within 0.5 km. In contrast, neither measure of Southern Leopard frog genetic diversity was associated with any habitat features at any scale we evaluated. Genetic diversity in the Dwarf salamander was strongly associated with land cover variables up to 2.5 km away from breeding wetlands, and/or results suggest that minimizing roads in wetland buffers may be beneficial to the maintenance of population genetic diversity. This study suggests that patterns of genetic differentiation and genetic diversity have associations with different habitat features across different spatial scales for two syntopic pond-breeding amphibian species.

A global database of nitrogen and phosphorus excretion rates of aquatic animals.

Animals can be important in modulating ecosystem-level nutrient cycling, although their importance varies greatly among species and ecosystems. Nutrient cycling rates of individual animals represent valuable data for testing the predictions of important frameworks such as the Metabolic Theory of Ecology (MTE) and ecological stoichiometry (ES). They also represent an important set of functional traits that may reflect both environmental and phylogenetic influences. Over the past two decades, studies of animal-mediated nutrient cycling have increased dramatically, especially in aquatic ecosystems. Here we present a global compilation of aquatic animal nutrient excretion rates. The dataset includes 10,534 observations from freshwater and marine animals of N and/or P excretion rates. These observations represent 491 species, including most aquatic phyla. Coverage varies greatly among phyla and other taxonomic levels. The dataset includes information on animal body size, ambient temperature, taxonomic affiliations, and animal body N:P. This data set was used to test predictions of MTE and ES, as described in Vanni and McIntyre (2016; Ecology DOI: 10.1002/ecy.1582).

The unseen invaders: introduced earthworms as drivers of change in plant communities in North American forests (a meta-analysis).

Globally, biological invasions can have strong impacts on biodiversity as well as ecosystem functioning. While less conspicuous than introduced aboveground organisms, introduced belowground organisms may have similarly strong effects. Here, we synthesize for the first time the impacts of introduced earthworms on plant diversity and community composition in North American forests. We conducted a meta-analysis using a total of 645 observations to quantify mean effect sizes of associations between introduced earthworm communities and plant diversity, cover of plant functional groups, and cover of native and non-native plants. We found that plant diversity significantly declined with increasing richness of introduced earthworm ecological groups. While plant species richness or evenness did not change with earthworm invasion, our results indicate clear changes in plant community composition: cover of graminoids and non-native plant species significantly increased, and cover of native plant species (of all functional groups) tended to decrease, with increasing earthworm biomass. Overall, these findings support the hypothesis that introduced earthworms facilitate particular plant species adapted to the abiotic conditions of earthworm-invaded forests. Further, our study provides evidence that introduced earthworms are associated with declines in plant diversity in North American forests. Changing plant functional composition in these forests may have long-lasting effects on ecosystem functioning.

Convergence of detrital stoichiometry predicts thresholds of nutrient-stimulated breakdown in streams.

Nutrient enrichment of detritus-based streams increases detrital resource quality for consumers and stimulates breakdown rates of particulate organic carbon (C). The relative importance of dissolved inorganic nitrogen (N) vs. phosphorus (P) for detrital quality and their effects on microbial- vs. detritivore-mediated detrital breakdown are poorly understood. We tested effects of experimental N and P additions on detrital stoichiometry (C:N, C:P) and total and microbial breakdown (i.e., with and without detritivorous shredders, respectively) of five detritus types (four leaf litter species and wood) with different initial C : nutrient content. We enriched five headwater streams continuously for two years at different relative availabilities of N and P and compared breakdown rates and detrital stoichiometry to pretreatment conditions. Total breakdown rates increased with nutrient enrichment and were predicted by altered detrital stoichiometry. Streamwater N and P, fungal biomass, and their interactions affected stoichiometry of detritus. Streamwater N and P decreased detrital C:N, whereas streamwater P had stronger negative effects on detrital C:P. Nutrient addition and fungal biomass reduced C:N by 70% and C:P by 83% on average after conditioning, compared to only 26% for C:N and 10% for C:P under pretreatment conditions. Detritus with lowest initial nutrient content changed the most and had greatest increases in total breakdown rates. Detrital stoichiometry was reduced and differences among detritus types were homogenized by nutrient enrichment. With enrichment, detrital nutrient content approached detritivore nutritional requirements and stimulated greater detritivore vs. microbial litter breakdown. We used breakpoint regression to estimate values of detrital stoichiometry that can potentially be used to indicate elevated breakdown rates. Breakpoint ratios for total breakdown were 41 (C:N) and 1518 (C:P), coinciding with total breakdown rates that were ~1.9 times higher when C:N or C:P fell below these breakpoints. Microbial and shredder-mediated breakdown rates both increased when C:N and C:P were reduced, suggesting that detrital stoichiometry is useful for predicting litter breakdown dominated by either microbial or shredder activity. Our results show strong effects of nutrient enrichment on detrital stoichiometry and offer a robust link between a potential holistic nutrient loading metric (decreased and homogenized detrital stoichiometry) and increased C loss from stream ecosystems.

Possible Role of Fish and Frogs as Paratenic Hosts of Dracunculus medinensis, Chad.

Copepods infected with Dracunculus medinensis larvae collected from infected dogs in Chad were fed to 2 species of fish and tadpoles. Although they readily ingested copepods, neither species of fish, Nile tilapia (Oreochromis niloticus) nor fathead minnow (Pimephalis promelas), were found to harbor Dracunculus larvae when examined 2-3 weeks later. Tadpoles ingested copepods much more slowly; however, upon examination at the same time interval, tadpoles of green frogs (Lithobates [Rana] clamitans) were found to harbor small numbers of Dracunculus larvae. Two ferrets (Mustela putorius furo) were fed fish or tadpoles that had been exposed to infected copepods. Only the ferret fed tadpoles harbored developing Dracunculus larvae at necropsy 70-80 days postexposure. These observations confirm that D. medinensis, like other species in the genus Dracunculus, can readily survive and remain infective in potential paratenic hosts, especially tadpoles.

Migratory monarchs wintering in California experience low infection risk compared to monarchs breeding year-round on non-native milkweed.

Long-distance migration can lower infection risk for animal populations by removing infected individuals during strenuous journeys, spatially separating susceptible age classes, or allowing migrants to periodically escape from contaminated habitats. Many seasonal migrations are changing due to human activities including climate change and habitat alteration. Moreover, for some migratory populations, sedentary behaviors are becoming more common as migrants abandon or shorten their journeys in response to supplemental feeding or warming temperatures. Exploring the consequences of reduced movement for host-parasite interactions is needed to predict future responses of animal pathogens to anthropogenic change. Monarch butterflies (Danaus plexippus) and their specialist protozoan parasite Ophryocystis elektroscirrha (OE) provide a model system for examining how long-distance migration affects infectious disease processes in a rapidly changing world. Annual monarch migration from eastern North America to Mexico is known to reduce protozoan infection prevalence, and more recent work suggests that monarchs that forego migration to breed year-round on non-native milkweeds in the southeastern and south central Unites States face extremely high risk of infection. Here, we examined the prevalence of OE infection from 2013 to 2016 in western North America, and compared monarchs exhibiting migratory behavior (overwintering annually along the California coast) with those that exhibit year-round breeding. Data from field collections and a joint citizen science program of Monarch Health and Monarch Alert showed that infection frequency was over nine times higher for monarchs sampled in gardens with year-round milkweed as compared to migratory monarchs sampled at overwintering sites. Results here underscore the importance of animal migrations for lowering infection risk and motivate future studies of pathogen transmission in migratory species affected by environmental change.

Color perception influences microhabitat selection of refugia and affects monitoring success for a cryptic anuran species.

Perceptual-biases are important for understanding an animal's natural history, identifying potential ecological traps, and for developing effective means to monitor individuals and populations. Despite research demonstrating anurans having a positive phototactic response towards blue colors, we do not yet understand if color cues are used functionally beyond sexual selection. The aim of our study was to determine if color cues are used in selecting microhabitat, and if anuran's blue-positive phototactic response could increase selection of artificial PVC refugia used to monitor cryptic camouflaging anuran species. We captured 32 Cope's Gray Treefrogs and placed them in mesh enclosures with three PVC tubes painted blue, brown, and white. Concurrently, we placed blue, brown, or unpainted white PVC tubes in stratified arrays around a treefrog breeding pond, and counted the number of occasions treefrogs occupied different colored PVC tubes. In the confined choice experiment, treefrogs selected blue tubes (48.3%) significantly more often than brown (28.5%) or white (23.2%) tubes. Our field experiment mirrored these findings (52.0% of capture events in blue, 29.0% in brown, and 19.0% in unpainted white tubes). Our results suggest color influences Cope's Gray Treefrog microhabitat selection, and they utilize color vision when choosing refugia. We demonstrate simple, small changes based on perceptual-biases can induce behaviors that may in turn have large impacts on sampling techniques used in monitoring and inventorying. Incorporating non-traditional physiological measures into animal inventorying and monitoring programs can be used in the future to improve conservation efforts.

Detrital stoichiometry as a critical nexus for the effects of streamwater nutrients on leaf litter breakdown rates.

Nitrogen (N) and phosphorus (P) concentrations are elevated in many freshwater systems, stimulating breakdown rates of terrestrially derived plant litter; however, the relative importance of N and P in driving litter breakdown via microbial and detritivore processing are not fully understood. Here, we determined breakdown rates of two litter species, Acer rubrum (maple) and Rhododendron maximum (rhododendron), before (PRE) and during two years (YR1, YR2) of experimental N and P additions to five streams, and quantified the relative importance of hypothesized factors contributing to breakdown. Treatment streams received a gradient of P additions (low to high soluble reactive phosphorus [SRP]; ~10-85 µg/L) crossed with a gradient of N additions (high to low dissolved inorganic nitrogen [DIN]; ~472-96 µg/L) to achieve target molar N:P ratios ranging from 128 to 2. Litter breakdown rates increased above pre-treatment levels by an average of 1.1-2.2x for maple, and 2.7-4.9x for rhododendron in YR1 and YR2. We used path analysis to compare fungal biomass, shredder biomass, litter stoichiometry (nutrient content as C:N or C:P), discharge, and streamwater temperature as predictors of breakdown rates and compared models containing streamwater N, P or N + P and litter C:N or C:P using model selection criteria. Litter breakdown rates were predicted equally with either streamwater N or P (R2 = 0.57). In models with N or P, fungal biomass, litter stoichiometry, discharge, and shredder biomass predicted breakdown rates; litter stoichiometry and fungal biomass were most important for model fit. However, N and P effects may have occurred via subtly different pathways. Litter N content increased with fungal biomass (N-driven effects) and litter P content increased with streamwater P availability (P-driven effects), presumably via P storage in fungal biomass. In either case, the effects of N and P through these pathways were associated with higher shredder biomass and breakdown rates. Our results suggest that N and P stimulate litter breakdown rates via mechanisms in which litter stoichiometry is an important nexus for associated microbial and detritivore effects.

Low-to-moderate nitrogen and phosphorus concentrations accelerate microbially driven litter breakdown rates.

Particulate organic matter (POM) processing is an important driver of aquatic ecosystem productivity that is sensitive to nutrient enrichment and.drives ecosystem carbon (C) loss. Although studies of single concentrations of nitrogen (N) or phosphorus (P) have shown effects at relatively low concentrations, responses of litter breakdown rates along gradients of low-to-moderate N and P concentrations are needed to establish likely interdependent effects of dual N and P enrichment on baseline activity in stream ecosystems. We established 25 combinations of dissolved inorganic N (DIN; 55-545 µg/L) and soluble reactive P (SRP; 4-86 µg/L) concentrations with corresponding N:P molar ratios of 2-127 in experimental stream channels. We excluded macroinvertebrates, focusing on microbially driven breakdown of maple (Acer rubrum L.) and rhododendron (Rhododendron maximum L.) leaf litter. Breakdown rates, k, per day (d-1) and per degree-day (dd-l), increased by up to 6X for maple and 12× for rhododendron over our N and P enrichment gradient compared to rates at low ambient N and P concentrations. The best models of k (d- and dd-1) included litter species identity and N and P concentrations; there was evidence for both additive and interactive effects of N and P. Models explaining variation in k dd-1 were supported by N and P for both maple and rhododendron (R =0.67 and 0.33, respectively). Residuals in the relationship between k dd-1 and N concentration were largely explained by P, but residuals for k dd-1 and P. concentration were less adequately explained by N. Breakdown rates were more closely related to nutrient concentrations than variables associated with measurements of two mechanistic parameters associated with C loss (fungal biomass and microbial respiration rate). We also determined the effects of nutrient addition on litter C: nutrient stoichiometry and found reductions in litter C:N and C:P along our experimental nutrient gradient. Our results indicate that microbially driven litter processing rates increase across low-to-moderate nutrient gradients that are now common throughout human-modified landscapes.

Variable infection of stream salamanders in the southern Appalachians by the trematode Metagonimoides oregonensis (family: Heterophyidae).

Many factors contribute to parasites varying in host specificity and distribution among potential hosts. Metagonimoides oregonensis is a digenetic trematode that uses stream-dwelling plethodontid salamanders as second intermediate hosts in the Eastern US. We completed a field survey to identify which stream salamander species, at a regional level, are most likely to be important for transmission to raccoon definitive hosts. We surveyed six plethodontid species (N = 289 salamanders) from 23 Appalachian headwater sites in North Carolina: Desmognathus quadramaculatus (n = 69), Eurycea wilderae (n = 160), Desmognathus ocoee (n = 31), Desmognathus monticola (n = 3), Eurycea guttolineata (n = 7), and Gyrinophilus porphyriticus (n = 19). We found infection in all species except D. monticola. Further analysis focused on comparing infection in the two most abundant species, D. quadramaculatus and E. wilderae. We found that D. quadramaculatus had significantly higher infection prevalence and intensity, probably due to a longer aquatic larval period and larger body sizes and thus greater cumulative exposure to the parasite.

Loss of migratory behaviour increases infection risk for a butterfly host.

Long-distance animal migrations have important consequences for infectious disease dynamics. In some cases, migration lowers pathogen transmission by removing infected individuals during strenuous journeys and allowing animals to periodically escape contaminated habitats. Human activities are now causing some migratory animals to travel shorter distances or form sedentary (non-migratory) populations. We focused on North American monarch butterflies and a specialist protozoan parasite to investigate how the loss of migratory behaviours affects pathogen spread and evolution. Each autumn, monarchs migrate from breeding grounds in the eastern US and Canada to wintering sites in central Mexico. However, some monarchs have become non-migratory and breed year-round on exotic milkweed in the southern US. We used field sampling, citizen science data and experimental inoculations to quantify infection prevalence and parasite virulence among migratory and sedentary populations. Infection prevalence was markedly higher among sedentary monarchs compared with migratory monarchs, indicating that diminished migration increases infection risk. Virulence differed among parasite strains but was similar between migratory and sedentary populations, potentially owing to high gene flow or insufficient time for evolutionary divergence. More broadly, our findings suggest that human activities that alter animal migrations can influence pathogen dynamics, with implications for wildlife conservation and future disease risks.

Salamander growth rates increase along an experimental stream phosphorus gradient.

Nutrient-driven perturbations to the resource base of food webs are predicted to attenuate with trophic distance, so it is unclear whether higher-level consumers will generally respond to anthropogenic nutrient loading. Few studies have tested whether nutrient (specifically, nitrogen [N] and phosphorus [P]) enrichment of aquatic ecosystems propagates through multiple trophic levels to affect predators, or whether N vs. P is relatively more important in driving effects on food webs. We conducted two-year whole-stream N and P additions to five streams to generate gradients in N and P concentration and N:P ratio (target N:P = 2, 8, 16, 32, 128). Larval salamanders are vertebrate predators of primary and secondary macroinvertebrate consumers in many heterotrophic headwater streams in which the basal resources are detritus and associated microorganisms. We determined the effects of N and P on the growth rates of caged and free-roaming larval Desmognathus quadramaculatus and the average body size of larval Eurycea wilderae. Growth rates and average body size increased by up to 40% and 60%, respectively, with P concentration and were negatively related to N:P ratio. These findings were consistent across both species of salamanders using different methodologies (cage vs. free-roaming) and at different temporal scales (3 months vs. 2 yr). Nitrogen concentration was not significantly related to increased growth rate or body size of the salamander species tested. Our findings suggest that salamander growth responds to the relaxation of ecosystem-level P limitation and that moderate P enrichment can have relatively large effects on vertebrate predators in detritus-based food webs.