Raoul K. Boughton

The Fungicide Chlorothalonil Is Nonlinearly Associated with Corticosterone Levels, Immunity, and Mortality in Amphibians

Background: Contaminants have been implicated in declines of amphibians, a taxon with vital systems similar to those of humans. However, many chemicals have not been thoroughly tested on amphibians or do not directly kill them. Objective: Our goal in this study was to quantify amphibian responses to chlorothalonil, the most commonly used synthetic fungicide in the United States. Methods: We reared Rana sphenocephala (southern leopard frog) and Osteopilus septentrionalis (Cuban treefrog) in outdoor mesocosms with or without 1 time (1×) and 2 times (2×) the expected environmental concentration (EEC) of chlorothalonil (~ 164 μg/L). We also conducted two dose–response experiments on O. septentrionalis, Hyla squirella (squirrel treefrog), Hyla cinerea (green treefrog), and R. sphenocephala and evaluated the effects of chlorothalonil on the stress hormone corticosterone. Results: For both species in the mesocosm experiment, the 1× and 2× EEC treatments were associated with > 87% and 100% mortality, respectively. In the laboratory experiments, the approximate EEC caused 100% mortality of all species within 24 hr; 82 μg/L killed 100% of R. sphenocephala, and 0.0164 μg/L caused significant tadpole mortality of R. sphenocephala and H. cinerea. Three species 
showed a nonmonotonic dose response, with low and high concentrations causing significantly greater mortality than did intermediate concentrations or control treatments. For O. septentrionalis, corticosterone exhibited a similar nonmonotonic dose response and chlorothalonil concentration was inversely associated with liver tissue and immune cell densities (< 16.4 μg/L). Conclusions: Chlorothalonil killed nearly every amphibian at the approximate EEC; at concentrations to which humans are commonly exposed, it increased mortality and was associated with elevated corticosterone levels and changes in immune cells. Future studies should directly quantify the effects of chlorothalonil on amphibian populations and human health.

Early-life exposure to a herbicide has enduring effects on pathogen-induced mortality

Exposure to stressors at formative stages in the development of wildlife and humans can have enduring effects on health. Understanding which, when and how stressors cause enduring health effects is crucial because these stressors might then be avoided or mitigated during formative stages to prevent lasting increases in disease susceptibility. Nevertheless, the impact of early-life exposure to stressors on the ability of hosts to resist and tolerate infections has yet to be thoroughly investigated. Here, we show that early-life, 6-day exposure to the herbicide atrazine (mean ± s.e.: 65.9±3.48 µg l−1) increased frog mortality 46 days after atrazine exposure (post-metamorphosis), but only when frogs were challenged with a chytrid fungus implicated in global amphibian declines. Previous atrazine exposure did not affect resistance of infection (fungal load). Rather, early-life exposure to atrazine altered growth and development, which resulted in exposure to chytrid at more susceptible developmental stages and sizes, and reduced tolerance of infection, elevating mortality risk at an equivalent fungal burden to frogs unexposed to atrazine. Moreover, there was no evidence of recovery from atrazine exposure. Hence, reducing early-life exposure of amphibians to atrazine could reduce lasting increases in the risk of mortality from a disease associated with worldwide amphibian declines. More generally, these findings highlight that a better understanding of how stressors cause enduring effects on disease susceptibility could facilitate disease prevention in wildlife and humans, an approach that is often more cost-effective and efficient than reactive medicine.

Association of ecotones with relative elevation and fire in an upland Florida landscape

Abstract Question: What are the importance of elevation and fire in maintaining ecotones of Florida scrub assemblages along a gradual topographic gradient? Location: Archbold Biological Station (ABS), 12 km south of Lake Placid, Florida, USA. Methods: Vegetation cover of upland Florida shrublands was quantified using the line-intercept method along 20 transects traversing similar elevation gradients, stratified by time since fire (TSF). We objectively identified shrubland ecotones using a split moving windows boundary analysis (SMW) with three different window widths. Non-metric multidimensional scaling ordination was used to determine relationships among plant assemblages defined by SMW. Results: We located up to four ecotones per transect, the majority of which were wide, highly heterogeneous zones. Relative elevation controlled the distribution of plant assemblages in upland Florida shrublands. Ecotones in shrublands > 30 years TSF had relatively low dissimilarity values in SMW, indicating that previously discrete plant assemblages with longer TSF were becoming more similar with time. Conclusions: Split Moving Windows (SMW) analysis identified ecotones relatively well although patches generated by oak clonal growth were sometimes identified as ecotones. Fire suppression caused ecotones to become more diffuse, suggesting that without fire at least every 30 years, discrete plant assemblages within upland Florida shrublands will be more continuous. Abbreviations: ABS = Archbold Biological Station; SMW = Split moving window; TSF =Time since fire; NMDS = Non-metric multi-dimensional scaling; OSF = Open Scrubby Flatwoods; RS = Rosemary Scrub; SF = Scrubby Flatwoods; F = Flatwoods; SCB = Squared city block dissimilarities. Nomenclature: Wunderlin (1998); Abrahamson et al. (1984).

Selection on innate immunity and body condition in Florida scrub-jays throughout an epidemic.

Opportunities to investigate selection in free-living species during a naturally occurring epidemic are rare; however, we assessed innate immunocompetence in Florida scrub-jays before the population suffered the greatest over-winter mortality in 20 years of study. Propitiously, three months prior to the epidemic, we had sampled a number of male breeders to evaluate a suite of physiological measures that are commonly used to estimate the overall health-state of an individual. There was a significant, positive selection gradient for both Escherichia coli bacterial killing capability and body condition, suggesting that directional selection had occurred upon each of these traits during the disease epidemic.

Age-related differences in baseline and stress-induced corticosterone in Florida scrub-jays

In physiological studies of free-living species, it is essential to consider the context of the life history stage at which an individual was observed in order to link measures of physiology with ecological parameters. One such measure that is important to consider is the age of an individual. We tested whether baseline or stress-induced corticosterone levels vary with age in free-living Florida scrub-jays (Aphelocoma coerulescens) during the pre-breeding period. Corticosterone (CORT), the primary avian stress hormone, is released in response to stressful stimuli, and stimulates gluconeogenesis; however, it also serves as a chemical messenger that can influence other physiological processes, reproduction, and behavior. We monitored both baseline CORT levels longitudinally throughout a five-year period and stress-induced CORT responses over a shorter two-year period. We predicted that older jays would have lower baseline CORT levels and a dampened stress response compared to younger birds, as has been shown in other avian species. We found no significant differences in baseline CORT levels with age. We found a decrease in total corticosterone responses to a stressor with age, however, the oldest birds in the population showed greater total corticosterone responses to a stressor. These results may be a product of age-related changes in physiological processes related to the stress response or a result of selection acting on the population, resulting in only the most responsive individuals surviving to old age.

Development of the adrenal stress response in the Florida scrub-jay (Aphelocoma coerulescens).

Nestlings of altricial species undergo a period of substantial growth and development in the nest after hatching. The hypothalamic-pituitary-adrenal axis regulates the release of stress hormones such as corticosterone, which in adults is critical in allowing an animal to respond to a stressor. However, activation of this axis in young birds may be detrimental to growth and possibly survival. The developmental hypothesis predicts that altricial nestlings should display a dampened corticosterone response to stress as a means of protection against the potentially harmful effects of elevated corticosterone. We examined this hypothesis in Florida scrub-jays, a cooperatively breeding species with altricial young. Blood samples were collected from nestlings, nutritionally independent young, and yearlings for measurement of corticosterone levels. Baseline corticosterone levels did not differ between age-classes; however, stress-induced corticosterone levels were highest in yearlings, intermediate in independent young, and lowest in nestlings. The nestling stress response was also of a shorter duration than the response in independent young and yearlings. This variation in stress responsiveness across ages may be an adaptive mechanism to protect the developing bird from the negative effects of corticosterone on growth and cognitive development.

Corticosterone administration does not affect timing of breeding in Florida scrub-jays (Aphelocoma coerulescens).

Providing supplemental food to Florida scrub-jays (Aphelocoma coerulescens) causes a reliable advance in clutch initiation of 1 to 2 weeks. In some years, supplemental food appeared to not only advance laying date but also decrease baseline concentrations of corticosterone (CORT) relative to controls. The coincidence of low CORT levels and early breeding led us to hypothesize that CORT serves to communicate information about environmental conditions to the hypothalamic-pituitary-gonadal (HPG) axis, which ultimately influences the timing of breeding. To test this hypothesis, we administered small oral doses of CORT three times each day to female breeders that were provisioned with supplemental food. We compared clutch initiation dates of the CORT-dosed females to females with supplementation but no exogenous CORT and to females with neither CORT nor supplemental food. CORT administration had a strong temporary effect on circulating CORT concentrations but clutch initiation did not differ between the two groups of supplemented birds, both of which laid eggs approximately 10 days earlier than nonsupplemented birds. Furthermore, during the year of our study we found no reduction in baseline CORT concentrations in our undosed supplemental groups, as had been observed in past studies.

ROAD EFFECTS ON FOOD AVAILABILITY AND ENERGETIC INTAKE IN FLORIDA SCRUB-JAYS (APHELOCOMA COERULESCENS)

ABSTRACT. Roads may have detrimental effects on many bird species, but areas adjacent to roads also provide habitat and food for some species. To address the influence of roadside habitat on the federally threatened Florida Scrub-Jay (Aphelocoma coerulescens), we compared foraging effort, the amount of food collected, and estimated energy intake in three habitat types: (1) edges along a paved road (roadside), (2) edges created by human-maintained habitat without a road (pasture), and (3) natural scrub-dominated areas with minimal human-maintained edges (interior). We used focal watches of male breeders during three stages of the breeding season (incubation, nestling, and fledged young) in two consecutive years to assess variation in food availability and energetic intake among the three habitat types. Florida Scrub-Jays in roadside habitats handled (consumed, cached, or fed to their mate or offspring) significantly more food items and spent less time foraging during the nestling stage than interior and pasture jays, but no differences in handled food were found among the three habitat types during the incubation and fledgling stages. Although they spent less time foraging, roadside jays had higher rates of energy intake than interior and pasture jays during the incubation stage. There were no differences in these measures of foraging during the incubation, nestling, and fledgling stages between interior and pasture jays. Our results suggest that roads or roadside habitat, not simply edge habitats, have a positive effect on food availability and energy intake in Florida Scrub-Jays, although the patterns differ across breeding stages.

Older can be better: physiological costs of paternal investment in the Florida scrub-jay

In species that undergo actuarial senescence, the value of current reproduction is predicted to increase relative to the value of future reproduction with age, as the probability of survival to another reproductive event is reduced. Therefore, life history theory predicts that aging animals should increase their investment in reproduction. However, an increase in reproductive investment may carry significant costs to the breeding individuals. We recorded provisioning rates of Florida scrub-jay male breeders, followed by their immediate capture to assess body condition and collect blood for an in vitro test of immunocompetence and an assay of baseline corticosterone for a measure of stress. Older males provisioned offspring and brooding mates at the highest rates. There was no evidence of any physiological deficits in males with high provisioning rates, independent of age. It appears that birds that survive to old age are high quality birds that maintain good physiological condition, which complements the value of experience and permits maximal investment in offspring.

Modification by an invasive ecosystem engineer shifts a wet prairie to a monotypic stand

At the landscape scale, ecosystem engineers are expected to increase species diversity; however, diversity could decline if the ecosystem engineer is over-abundant. Thus, invasive ecosystem engineers are expected to have strong impacts, due to their high abundances and novel disturbances. An invasive ecosystem engineer, the feral swine (Sus scrofa), is a species that creates intense soil disturbances, altering soil and plant communities. In this study, we examine the effects of this invasive ecosystem engineer on experimental plant plots that had been protected for over a decade. Feral swine avoided recently burned plots and preferred plots with N addition. Rooted plots shifted from a bunchgrass dominated wet prairie to a monotypic stand of the native, Lachnanthes caroliana. Feral swine were also attracted to plots with existing patches of L. caroliana suggesting a potential positive feedback between swine activity and L. caroliana patch expansion that could result in an alternative state.