Effects of Lead (Pb) from Smelter Operations in an Urban Terrestrial Food Chain at a Colorado Superfund Site.

Lead (Pb) is ubiquitous in urban environments, and it is a risk factor for wildlife. But wildlife are particularly at risk for exposure near smelters in urban areas where higher than safe Pb levels in the soil have the potential to transfer to the food chain. Therefore, we investigated whether wildlife are at risk of Pb exposure and differences in Pb bioaccumulation in trophic levels at a Superfund site in an urban area of Colorado. We sampled soil, vegetation, arthropods, and birds at four sites: two contaminated sites (one at the Superfund site and one near the Superfund site) and two reference sites with low predicted Pb contamination. We found significantly higher Pb levels in the soil at the contaminated Slag pile at the Superfund site, compared to the other sites. At the Slag pile, Pb levels were the highest in vegetation and both arthropods and birds accumulated Pb. However, Pb did not increase between trophic levels in the terrestrial food web at the site. We concluded that smelter operations at the Superfund site resulted in significantly higher levels of Pb in the soil and sublethal accumulation in all taxa studied. This research can be used to mitigate the risks to vulnerable wildlife populations exposed to Pb at the Superfund site. Further examination of Pb toxicity in vulnerable taxa is critical to conserving wild populations and ecosystems near contamination zones.

Correlates of bird collisions with buildings across three North American countries.

Collisions with buildings cause up to 1 billion bird fatalities annually in the United States and Canada. However, efforts to reduce collisions would benefit from studies conducted at large spatial scales across multiple study sites with standardized methods and consideration of species- and life-history-related variation and correlates of collisions. We addressed these research needs through coordinated collection of data on bird collisions with buildings at sites in the United States (35), Canada (3), and Mexico (2). We collected all carcasses and identified species. After removing records for unidentified carcasses, species lacking distribution-wide population estimates, and species with distributions overlapping fewer than 10 sites, we retained 269 carcasses of 64 species for analysis. We estimated collision vulnerability for 40 bird species with ≥2 fatalities based on their North American population abundance, distribution overlap in study sites, and sampling effort. Of 10 species we identified as most vulnerable to collisions, some have been identified previously (e.g., Black-throated Blue Warbler [Setophaga caerulescens]), whereas others emerged for the first time (e.g., White-breasted Nuthatch [Sitta carolinensis]), possibly because we used a more standardized sampling approach than past studies. Building size and glass area were positively associated with number of collisions for 5 of 8 species with enough observations to analyze independently. Vegetation around buildings influenced collisions for only 1 of those 8 species (Swainson's Thrush [Catharus ustulatus]). Life history predicted collisions; numbers of collisions were greatest for migratory, insectivorous, and woodland-inhabiting species. Our results provide new insight into the species most vulnerable to building collisions, making them potentially in greatest need of conservation attention to reduce collisions and into species- and life-history-related variation and correlates of building collisions, information that can help refine collision management.

Correlaciones de las Colisiones de Aves contra Edificios en Tres Países de América del Norte Resumen Las colisiones contra los edificios causan hasta mil millones de fatalidades de aves al año en los Estados Unidos y en Canadá. Sin embargo, los esfuerzos por reducir estas colisiones se beneficiarían con estudios realizados a grandes escalas espaciales en varios sitios de estudio con métodos estandarizados y considerando las variaciones relacionadas a la historia de vida y a la especie y las correlaciones de las colisiones. Abordamos estas necesidades de investigación por medio de una recolección coordinada de datos sobre las colisiones de aves contra edificios en los Estados Unidos (35), Canadá (3) y México (2). Recolectamos todos los cadáveres y los identificamos hasta especie. Después de retirar los registros de cadáveres no identificados, las especies sin estimaciones poblacionales a nivel distribución y las especies con distribuciones traslapadas en menos de diez sitios, nos quedamos con 269 cadáveres de 64 especies para el análisis. Estimamos la vulnerabilidad a colisiones para 40 especies con ≥2 fatalidades con base en la abundancia poblacional para América del Norte, el traslape de su distribución entre los sitios de estudio y el esfuerzo de muestreo. De las diez especies que identificamos como las más vulnerables a las colisiones, algunas han sido identificadas previamente (Setophaga caerulescens), y otras aparecieron por primera vez (Sitta carolinensis), posiblemente debido a que usamos una estrategia de muestreo más estandarizada que en los estudios previos. El tamaño del edificio y el área del vidrio estuvieron asociados positivamente con el número de colisiones para cinco de ocho especies con suficientes observaciones para ser analizadas independientemente. La vegetación alrededor de los edificios influyó sobre las colisiones solamente para una de esas ocho especies Catharus ustulatus). Las historias de vida pronosticaron las colisiones; el número de colisiones fue mayor para las especies migratorias, insectívoras y aquellas que habitan en las zonas boscosas. Nuestros resultados proporcionan una nueva perspectiva hacia las especies más vulnerables a las colisiones contra edificios, lo que las pone en una necesidad potencialmente mayor de atención conservacionista para reducir estas colisiones y de estudio de las variaciones relacionadas con la especie y la historia de vida y las correlaciones de las colisiones contra edificios, información que puede ayudar a refinar el manejo de colisiones.

Sexually selected traits as bioindicators: exposure to mercury affects carotenoid-based male bill color in zebra finches.

To examine whether sexually selected traits are particularly sensitive bioindicators of environmental toxicants, we assessed the effects of exposure to environmentally relevant dietary concentrations of the pollutant methylmercury on pigment coloration in zebra finches (Taeniopygia guttata). First, we tested whether effects of methylmercury on coloration were influenced by timing of exposure. Birds were either exposed developmentally (up to 114 days after hatching), as adults (after reaching sexual maturity), or for their entire life. Bill coloration, which is a carotenoid-based, sexually selected trait, was less red in males with lifetime exposure to methylmercury, compared to controls. Neither adult, nor developmental exposure influenced bill color in adult males, with the possible exception of early exposure of nestlings. Among females, where bill color is not under strong sexual selection, neither lifetime nor adult exposure to methylmercury affected bill color. For males and females, there was no effect of either lifetime or adult methylmercury exposure on coloration of back feathers, which is a non-sexually-dimorphic, melanin-based trait that is not likely the result of sexual selection. This study is a comprehensive experimental test of the proposal that sexually selected traits may be particularly useful bioindicators of the stress imposed by environmental toxins such as methylmercury.

Mercury delays cerebellar development in a model songbird species, the zebra finch.

Mercury exposure can disrupt development of the cerebellum, part of the brain essential for coordination of movement through a complex environment, including flight. In precocial birds, such as fowl, the cerebellum develops embryonically, and the chick is capable of leaving the nest within hours of hatching. However, most birds, including all songbirds, are altricial, and spend weeks in the nest between hatching and fledging. The objective of this study was to describe the normal development of the cerebellum in a model altricial songbird so as to determine the effect of exposure to mercury on cerebellar maturation. Adult zebra finch (Taeniopygia guttata) pairs were fed either a control diet, or a diet augmented with one of four treatment-levels of methylmercury (0.3-2.4 µg/g wet weight), and their offspring, the subjects of this study, were fed the same diet by parents. We documented, for the first time, the schedule of cerebellar development in an altricial bird, and compared stages of development among methylmercury-exposed groups. For all treatments of methylmercury, the age of completion of cellular migration was later than for control zebra finches, indicating a delay in cerebellar maturation. Displaced (heterotopic) Purkinje neurons, a pathology typical of methylmercury exposure in developing vertebrate brains, were more numerous in methylmercury-exposed birds, and persisted at least until the age of independence. Delays in maturation of the cerebellum could delay fledging in altricial bird species, with potential serious implications for the fitness of exposed individuals, as predation rates in the nest are often very high.

Exposure to Lead (Pb2+) Eliminates Avoidance of Pb-Treated Oviposition Substrates in a Dose-Dependent Manner in Female Vinegar Flies.

Female vinegar flies (Drosophila melanogaster) preferentially oviposit eggs on oviposition substrates that decrease larval foraging costs. We tested whether female D. melanogaster would avoid oviposition substrates containing lead (Pb2+), which could potentially decrease offspring fitness. Wild type D. melanogaster were reared on control or Pb-treated medium from egg stage to adulthood and tested for differences in oviposition substrate preference, fecundity (number of eggs laid) and Pb accumulation. Control females laid a significantly lower proportion of eggs on Pb-treated substrates than Pb-treated females. Pb-treated females laid significantly more eggs than control females. Pb-treated adults accumulated significantly more Pb than control-treated adults. These results indicate that Pb exposure disrupts normal oviposition avoidance behaviors, which could increase larval foraging costs for larval offspring. These factors could induce population declines and have cascading implications for the ecosystem.

Explaining variation in Colorado songbird blood mercury using migratory behavior, foraging guild, and diet.

Methylmercury is a contaminant of growing global concern that has been shown to accumulate in a variety of taxa, including songbirds. Birds in the same area can accumulate mercury to strikingly different levels. While diet and trophic level clearly play an important role in mercury bioaccumulation and biomagnification, other factors including foraging guilds and migratory behavior may influence mercury levels as well. Here we examine interspecific variation in blood mercury levels in songbirds living in the Fountain Creek watershed on the Front Range of Colorado. We found that the species with the highest mercury had blood mercury concentrations over 75 times higher than the species with the lowest levels. Carnivores had the highest blood mercury levels, but ground foraging and long distance migration also were correlated with higher mercury concentrations. This information may shed light on what species are most at risk from mercury pollution and help to target conservation resources at contaminated sites.

Multiple hypotheses explain variation in extra-pair paternity at different levels in a single bird family.

Extra-pair paternity (EPP), where offspring are sired by a male other than the social male, varies enormously both within and among species. Trying to explain this variation has proved difficult because the majority of the interspecific variation is phylogenetically based. Ideally, variation in EPP should be investigated in closely related species, but clades with sufficient variation are rare. We present a comprehensive multifactorial test to explain variation in EPP among individuals in 20 populations of nine species over 89 years from a single bird family (Maluridae). Females had higher EPP in the presence of more helpers, more neighbours or if paired incestuously. Furthermore, higher EPP occurred in years with many incestuous pairs, populations with many helpers and species with high male density or in which males provide less care. Altogether, these variables accounted for 48% of the total and 89% of the interspecific and interpopulation variation in EPP. These findings indicate why consistent patterns in EPP have been so challenging to detect and suggest that a single predictor is unlikely to account for the enormous variation in EPP across levels of analysis. Nevertheless, it also shows that existing hypotheses can explain the variation in EPP well and that the density of males in particular is a good predictor to explain variation in EPP among species when a large part of the confounding effect of phylogeny is excluded.

Form of Dietary Methylmercury does not Affect Total Mercury Accumulation in the Tissues of Zebra Finch.

Exposure to mercury in humans, other mammals, and birds is primarily dietary, with mercury in the methylated form and bound to cysteine in the tissues of prey items. Yet dosing studies are generally carried out using methylmercury chloride. Here we tested whether the accumulation of total mercury in zebra finch blood, egg, muscle, liver, kidney or brain differed depending on whether dietary mercury was complexed with chloride or cysteine. We found no effect of form of mercury on tissue accumulation. Some previous studies have found lower accumulation of mercury in tissues of animals fed complexed mercury. Much remains to be understood about what happens to ingested mercury once it enters the intestines, but our results suggest that dietary studies using methylmercury chloride in birds will produce similar tissue accumulation levels to those using methylmercury cysteine.

Integrative behavioral ecotoxicology: bringing together fields to establish new insight to behavioral ecology, toxicology, and conservation.

The fields of behavioral ecology, conservation science, and environmental toxicology individually aim to protect and manage the conservation of wildlife in response to anthropogenic stressors, including widespread anthropogenic pollution. Although great emphasis in the field of toxicology has been placed on understanding how single pollutants affect survival, a comprehensive, interdisciplinary approach that includes behavioral ecology is essential to address how anthropogenic compounds are a risk for the survival of species and populations in an increasingly polluted world. We provide an integrative framework for behavioral ecotoxicology using Tinbergen's four postulates (causation and mechanism, development and ontogeny, function and fitness, and evolutionary history and phylogenetic patterns). The aims of this review are: 1) to promote an integrative view and re-define the field of integrative behavioral ecotoxicology; 2) to demonstrate how studying ecotoxicology can promote behavior research; and 3) to identify areas of behavioral ecotoxicology that require further attention to promote the integration and growth of the field.

Blood Mercury Levels of Zebra Finches Are Heritable: Implications for the Evolution of Mercury Resistance.

Mercury is a ubiquitous metal contaminant that negatively impacts reproduction of wildlife and has many other sub-lethal effects. Songbirds are sensitive bioindicators of mercury toxicity and may suffer population declines as a result of mercury pollution. Current predictions of mercury accumulation and biomagnification often overlook possible genetic variation in mercury uptake and elimination within species and the potential for evolution in affected populations. We conducted a study of dietary mercury exposure in a model songbird species, maintaining a breeding population of zebra finches (Taeniopygia guttata) on standardized diets ranging from 0.0-2.4 µg/g methylmercury. We applied a quantitative genetics approach to examine patterns of variation and heritability of mercury accumulation within dietary treatments using a method of mixed effects modeling known as the 'animal model'. Significant variation in blood mercury accumulation existed within each treatment for birds exposed at the same dietary level; moreover, this variation was highly repeatable for individuals. We observed substantial genetic variation in blood mercury accumulation for birds exposed at intermediate dietary concentrations. Taken together, this is evidence that genetic variation for factors affecting blood mercury accumulation could be acted on by selection. If similar heritability for mercury accumulation exists in wild populations, selection could result in genetic differentiation for populations in contaminated locations, with possible consequences for mercury biomagnification in food webs.

Effect of laying sequence on egg mercury in captive zebra finches: an interpretation considering individual variation.

Bird eggs are used widely as noninvasive bioindicators for environmental mercury availability. Previous studies, however, have found varying relationships between laying sequence and egg mercury concentrations. Some studies have reported that the mercury concentration was higher in first-laid eggs or declined across the laying sequence, whereas in other studies mercury concentration was not related to egg order. Approximately 300 eggs (61 clutches) were collected from captive zebra finches dosed throughout their reproductive lives with methylmercury (0.3 µg/g, 0.6 µg/g, 1.2 µg/g, or 2.4 µg/g wet wt in diet); the total mercury concentration (mean ± standard deviation [SD] dry wt basis) of their eggs was 7.03 ± 1.38 µg/g, 14.15 ± 2.52 µg/g, 26.85 ± 5.85 µg/g, and 49.76 ± 10.37 µg/g, respectively (equivalent to fresh wt egg mercury concentrations of 1.24 µg/g, 2.50 µg/g, 4.74 µg/g, and 8.79 µg/g). The authors observed a significant decrease in the mercury concentration of successive eggs when compared with the first egg and notable variation between clutches within treatments. The mercury level of individual females within and among treatments did not alter this relationship. Based on the results, sampling of a single egg in each clutch from any position in the laying sequence is sufficient for purposes of population risk assessment, but it is not recommended as a proxy for individual female exposure or as an estimate of average mercury level within the clutch.

PHA-stimulated immune-responsiveness in mercury-dosed zebra finches does not match results from environmentally exposed songbirds.

Dietary mercury exposure is associated with suppressed immune responsiveness in birds. This study examined the immune-responsiveness of domestic zebra finches (Taeniopygia guttata) experimentally exposed to mercury through their diet. We used the phytohemagglutinin (PHA) skin-swelling test to assay the effect of two modes of mercury exposure. Some finches received exposure to mercury only after reaching sexual maturity, while others were maintained on a mercury-dosed diet throughout life, including development. Each bird received one of five dietary concentrations of methylmercury cysteine (0.0, 0.3, 0.6, 1.2 or 2.4 ppm). In contrast to a study on wild songbirds at a mercury-contaminated site, we detected no relationship between mercury level and immunological response to PHA, regardless of mode of exposure. This result represents the first major difference found by our laboratory between wild birds exposed to environmental mercury and captive birds experimentally exposed to mercury.

The effect of mercury on baseline corticosterone in a breeding songbird.

Although songbirds accumulate mercury at rates equivalent to better-studied aquatic avian species, effects of mercury bioaccumulation in songbirds remain understudied. Little is known about the effects of mercury on endocrine physiology, but recent evidence indicates that mercury may disrupt the function of the hypothalamic-pituitary-adrenal axis. Both field-based correlational studies and a recent dosing experiment suggest that mercury exposure alters levels of the primary avian stress hormone, CORT. We sampled zebra finches that had been dosed with 0, 0.5, or 1.0 ppm dietary methylmercury for baseline CORT twice; once during pairing and once after successfully fledging young. Circulating levels of CORT were not significantly affected by mercury exposure. However, our findings indicate potentially important differences in CORT responses between the sexes when exposed to environmentally relevant doses of mercury across the nesting cycle.

Oxidative stress in songbirds exposed to dietary methylmercury.

Long-term, sublethal methylmercury exposure can cause reproductive depression, immune suppression, endocrine disruption and other problems in birds. We used two biomarkers to detect oxidative stress in livers of zebra finches (Taeniopygia guttata) developmentally exposed to sublethal levels of dietary methylmercury (0.0, 0.3, 0.6, 1.2, or 2.4 µg/g wet weight in diet). Our findings indicate that young adult finches exposed to environmentally relevant concentrations of mercury in ovo and through their diets, exhibited oxidative stress in their livers. We measured the ratio of the antioxidant glutathione in its reduced form (GSH) versus its oxidized form (GSSG) and the activity of the superoxide dismutase (SOD) enzyme suite. Blood total mercury served as a proxy for liver mercury concentration, and was on average 8.4 times the dietary dose (e.g., birds consuming 0.6 µg/g had blood mercury levels of ~5 µg/g on a wet weight basis). Consistent with what is known from large, aquatic bird species, there was a significant, negative relationship between GSH/GSSG ratios and tissue mercury concentrations, which is indicative of oxidative stress. This relationship was driven by a significant increase in the oxidized glutathione in the livers of birds with higher blood mercury levels. SOD activity was also found to have a significant, negative relationship with blood mercury.

Mercury reduces avian reproductive success and imposes selection: an experimental study with adult- or lifetime-exposure in zebra finch.

Mercury is a global pollutant that biomagnifies in food webs, placing wildlife at risk of reduced reproductive fitness and survival. Songbirds are the most diverse branch of the avian evolutionary tree; many are suffering persistent and serious population declines and we know that songbirds are frequently exposed to mercury pollution. Our objective was to determine the effects of environmentally relevant doses of mercury on reproductive success of songbirds exposed throughout their lives or only as adults. The two modes of exposure simulated philopatric species versus dispersive species, and are particularly relevant because of the heightened mercury-sensitivity of developing nervous systems. We performed a dosing study with dietary methylmercury in a model songbird species, the zebra finch (Taeniopygia guttata), at doses from 0.3 - 2.4 parts per million. Birds were exposed to mercury either as adults only or throughout their lives. All doses of mercury reduced reproductive success, with the lowest dose reducing the number of independent offspring produced in one year by 16% and the highest dose, representing approximately half the lethal dose for this species, causing a 50% reduction. While mercury did not affect clutch size or survivorship, it had the most consistent effect on the proportion of chicks that fledged from the nest, regardless of mode of exposure. Among birds exposed as adults, mercury caused a steep increase in the latency to re-nest after loss of a clutch. Birds exposed for their entire lifetimes, which were necessarily the offspring of dosed parents, had up to 50% lower reproductive success than adult-exposed birds at low doses of methylmercury, but increased reproductive success at high doses, suggesting selection for mercury tolerance at the highest level of exposure. Our results indicate that mercury levels in prey items at contaminated sites pose a significant threat to populations of songbirds through reduced reproductive success.

Lifelong exposure to methylmercury disrupts stress-induced corticosterone response in zebra finches (Taeniopygia guttata).

Mercury can disrupt the endocrine systems of mammals and fish, but little is known about its effects on avian hormones. The authors employed an experimental manipulation to show that methylmercury suppresses the stress-induced corticosterone response in birds, an effect previously unreported in the literature. Corticosterone regulates many normal metabolic processes, such as the maintenance of proper blood glucose levels during stressful daily fasting; an inability to increase corticosterone levels in response to stressors renders a bird less able to face a wide array of environmental challenges. The authors studied reproductively mature zebra finches that had been exposed to 0.0 µg/g, 0.3 µg/g, 0.6 µg/g, 1.2 µg/g, or 2.4 µg/g (wet wt) dietary methylmercury throughout their life (i.e., from the egg onward). In contrast to some field studies, the present study found no significant change in baseline plasma corticosterone concentrations attributable to chronic methylmercury exposure. However, a comparison between the baseline corticosterone levels and levels after 30 min of handling stress revealed that the ability of birds to mount a stress response was reduced with increasing blood total mercury concentration. These results are consistent with adrenal corticoid disruption caused by chronic mercury exposure and mirror a similar study on free-living nestling songbirds exposed to environmental mercury.

Familial differences in the effects of mercury on reproduction in zebra finches.

Ecotoxicologists often implicitly assume that populations are homogenous entities in which all individuals have similar responses to a contaminant. However, genetically variable responses occur within populations. This variation can be visualized using dose-response curves of genetically related groups, similar to the way that evolutionary biologists construct reaction norms. We assessed the variation in reproductive success of full-sibling families of captive zebra finches (Taeniopygia guttata) experimentally exposed to methylmercury. We found significant variation among families in the effects of methylmercury on several reproductive parameters. This variation suggests that there may be strong responses to selection for resistant genotypes in contaminated areas. This has important implications for the evolution of tolerance as well as risk assessment and wildlife conservation efforts on sites with legacy contamination.

Embryological staging of the Zebra Finch, Taeniopygia guttata.

Zebra Finches (Taeniopygia guttata) are the most commonly used laboratory songbird species, yet their embryological development has been poorly characterized. Most studies to date apply Hamburger and Hamilton stages derived from chicken development; however, significant differences in development between precocial and altricial species suggest that they may not be directly comparable. We provide the first detailed description of embryological development in the Zebra Finch under standard artificial incubation. These descriptions confirm that some of the features used to classify chicken embryos into stages are not applicable in an altricial bird such as the Zebra Finch. This staging protocol will help to standardize future studies of embryological development in the Zebra Finch.

Decreased immune response in zebra finches exposed to sublethal doses of mercury.

Mercury (Hg) is a ubiquitous contaminant with deleterious effects on many wildlife species. Most studies to date have focused on fish-eating birds and mammals because much historical Hg pollution is aquatic. Recently, however, comparable blood-Hg levels have been found in terrestrial insectivorous songbirds. As a result, research is needed to clarify the effects of Hg exposure on songbirds. One fundamental end point that is still poorly understood is the effect of Hg on the songbird immune system. If Hg affects the functioning of the immune system, exposed songbirds may be less able to mount an appropriate immune response against invading pathogens. To gain insight into how Hg affects songbird immune function on a cellular level, a flow cytometric assay was developed to measure lipopolysaccharide-induced B-lymphocyte proliferation in zebra finches (Taeniopygia guttata). This is the first experimental (dosing) study of the potential effect of Hg on songbird immune system functioning. Decreased B cell proliferation was observed after lipopolysaccharide exposure in individuals with greater concentrations of Hg in their blood and tissues. In addition, these individuals had decreased ratios of proliferating-to-resting B cells. This decrease in lymphocyte proliferation in response to an effective mitogen suggests that environmental exposure to sublethal levels of Hg may inhibit or delay B cell proliferation in songbirds, potentially increasing susceptibility to disease and decreasing survivorship.

Mercury levels of Nelson's and saltmarsh sparrows at wintering grounds in Virginia, USA.

Nelson's and saltmarsh sparrows (Ammodramus nelsoni and A. caudacutus) have recently been recognized as separate species, and because of their limited distributions and the susceptibility of their wetland habitats to climate change, these two new species are of conservation concern. Both species are known to bioaccumulate mercury at breeding sites in New England, USA where their ranges overlap, with the saltmarsh sparrow reported to have twice the concentration of blood total mercury. In this study we sampled both species on their shared wintering grounds, and documented that mercury exposure is lower than that reported for the breeding range, with saltmarsh sparrow blood mercury 2.6 times higher than in Nelson's sparrow. Feather mercury, which is incorporated on the breeding grounds, confirmed that saltmarsh sparrows had incorporated 2.3 times more mercury than Nelson's sparrows during the previous breeding season. A comparison of stable isotopes of nitrogen and carbon suggests that the higher exposure of saltmarsh sparrows may be not due to feeding at a higher trophic level, as previously hypothesized, but rather could be related to a difference in the carbon source at the base of each species' food chain. This study, along with recently published data from both species on additional breeding and wintering grounds, provides a more complete picture of relative mercury exposure. Saltmarsh sparrows are exposed to mercury levels that warrant concern, with the highest exposure being during the breeding season. Areas set aside for the long-term conservation of this species should be carefully assessed for mercury bioaccumulation.