Among-population variation in telomere regulatory proteins and their potential role as hidden drivers of intraspecific variation in life history.

Biologists aim to explain patterns of growth, reproduction and ageing that characterize life histories, yet we are just beginning to understand the proximate mechanisms that generate this diversity. Existing research in this area has focused on telomeres but has generally overlooked the telomere's most direct mediator, the shelterin protein complex. Shelterin proteins physically interact with the telomere to shape its shortening and repair. They also regulate metabolism and immune function, suggesting a potential role in life history variation in the wild. However, research on shelterin proteins is uncommon outside of biomolecular work. Intraspecific analyses can play an important role in resolving these unknowns because they reveal subtle variation in life history within and among populations. Here, we assessed ecogeographic variation in shelterin protein abundance across eight populations of tree swallow (Tachycineta bicolor) with previously documented variation in environmental and life history traits. Using the blood gene expression of four shelterin proteins in 12-day-old nestlings, we tested the hypothesis that shelterin protein gene expression varies latitudinally and in relation to both telomere length and life history. Shelterin protein gene expression differed among populations and tracked non-linear variation in latitude: nestlings from mid-latitudes expressed nearly double the shelterin mRNA on average than those at more northern and southern sites. However, telomere length was not significantly related to latitude. We next assessed whether telomere length and shelterin protein gene expression correlate with 12-day-old body mass and wing length, two proxies of nestling growth linked to future fecundity and survival. We found that body mass and wing length correlated more strongly (and significantly) with shelterin protein gene expression than with telomere length. These results highlight telomere regulatory shelterin proteins as potential mediators of life history variation among populations. Together with existing research linking shelterin proteins and life history variation within populations, these ecogeographic patterns underscore the need for continued integration of ecology, evolution and telomere biology, which together will advance understanding of the drivers of life history variation in nature.

Mercury causes degradation of spatial cognition in a model songbird species.

Mercury is a widespread pollutant of increasing global concern that exhibits a broad range of deleterious effects on organisms, including birds. Because the developing brain is well-known to be particularly vulnerable to the neurotoxic insults of mercury, many studies have focused on developmental effects such as on the embryonic brain and resulting behavioral impairment in adults. It is not well understood how the timing of exposure, for example exclusively in ovo versus throughout life, influences the impact of mercury. Using dietary exposure to environmentally relevant methylmercury concentrations, we examined the role that timing and duration of exposure play on spatial learning and memory in a model songbird species, the domesticated zebra finch (Taeniopygia guttata castanotis). We hypothesized that developmental exposure was both necessary and sufficient to disrupt spatial memory in adult finches. We documented profound disruption of memory for locations of hidden food at two spatial scales, cage- and room-sized enclosures, but found that both developmental and ongoing adult exposure were required to exhibit this behavioral impairment. Methylmercury-exposed birds made more mistakes before mastering the spatial task, because they revisited unrewarded locations repeatedly even after discovering the rewarded location. Contrary to our prediction, hippocampal volume was not affected in birds exposed to methylmercury over their lifetimes. The disruption of spatial cognition that we detected is severe and would likely have implications for survival and reproduction in wild birds; however, it appears that individuals that disperse or migrate from a contaminated site might recover later in life if no longer exposed to the toxicant.

Synthesis of Maternal Transfer of Mercury in Birds: Implications for Altered Toxicity Risk.

Maternal transfer is a predominant route of methylmercury (MeHg) exposure to offspring. We reviewed and synthesized published and unpublished data on maternal transfer of MeHg in birds. Using paired samples of females' blood (n = 564) and their eggs (n = 1814) from 26 bird species in 6 taxonomic orders, we conducted a meta-analysis to evaluate whether maternal transfer of MeHg to eggs differed among species and caused differential toxicity risk to embryos. Total mercury (THg) concentrations in eggs increased with maternal blood THg concentrations; however, the proportion of THg transferred from females to their eggs differed among bird taxa and with maternal THg exposure. Specifically, a smaller proportion of maternal THg was transferred to eggs with increasing female THg concentrations. Additionally, the proportion of THg that was transferred to eggs at the same maternal blood THg concentration differed among taxonomic orders, with waterfowl (Anseriformes) transferring up to 382% more THg into their eggs than songbirds (Passeriformes). We provide equations to predict THg concentrations in eggs using female blood THg concentrations, and vice versa, which may help translate toxicity benchmarks across tissues and life stages. Our results indicate that toxicity risk of MeHg can vary among bird taxa due to differences in maternal transfer of MeHg to offspring.

The impact of mercury on North American songbirds: effects, trends, and predictive factors.

Researchers were asked to contribute new results addressing questions about the exposure and effects of mercury (Hg) in North American songbirds, a rapidly declining group of species that is the subject of enduring interest for millions of birdwatchers, the general public and conservation scientists. Important questions to be answered include: Is Hg causing or exacerbating songbird population declines? Which North American songbirds are at most risk and in which landscapes? Are there aspects of songbird natural history that pre-dispose them to risks of Hg exposure and effects, in particular, their migratory behavior? In all, 61 authors contributed 15 studies addressing aspects of these questions. Articles in this special issue address an array of topics including: (1) three studies on health effects in the laboratory using a domesticated songbird model species, the zebra finch; (2) three studies on changes in songbird exposure to Hg over time spans from less than a decade to more than a century; (3) five studies on landscape characteristics or management practices that cause the oft-noted spatial variation in Hg accumulation by resident songbirds, from the subarctic tundra to high-elevation tropical forests; (4) three papers examining the recently recognized role of migration behavior in predicting risk to songbirds from Hg; and (5) one paper on the potential pitfalls of using feather Hg concentration as a bioindicator for Hg exposure. In summary, although there are many questions still to be answered, it is clear that the effects of Hg are persistent long after exposure, Hg exposure of North American songbirds is not improving, predicting exposure requires a detailed understanding of ecosystem processes beyond simply the amount of Hg present at a site, migration behavior predisposes songbirds to risk of Hg exposure and effects, and carefully selecting appropriate bioindicator sites, species, and tissues is critical to any monitoring efforts.

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 Dietary Methyl-Mercury Solely during Embryonic and Juvenile Development Halves Subsequent Reproductive Success in Adult Zebra Finches.

Long-term exposure to methyl-mercury has deleterious effects on avian reproduction. However, little is known about whether exposure to mercury solely during embryonic and juvenile development can have long-lasting effects on subsequent reproductive performance as adults. Birds that hatch on contaminated sites but disperse elsewhere will be exposed only during development. Hence, it is important to understand the reproductive consequences of avian exposure to methyl-mercury during early life. Accordingly, in this experiment, domesticated zebra finches ( Taeniopygia guttata) were exposed to dietary methyl-mercury (1.20 µg/g wet weight) from conception through independence (50 days post-hatching). Following maturity, developmentally exposed and control finches were paired within treatment groups and allowed to breed repeatedly for 8 months. Developmentally exposed pairs hatched 32% fewer eggs and produced 50% fewer independent juveniles despite transferring only traces of mercury into their offspring. This is the first example of mercury-related reproductive declines in birds not exposed to mercury during breeding. The magnitude of reproductive decline was similar to that of zebra finches exposed to methyl-mercury during the breeding process. Bird populations breeding in contaminated habitats may suffer from a 2-fold fitness cost of mercury exposure; adult exposure compromises parents' reproduction, while offspring exposure results in reduced reproduction in the next generation.

Impacts of Sublethal Mercury Exposure on Birds: A Detailed Review.

Mercury is a ubiquitous environmental contaminant known to accumulate in, and negatively affect, fish-eating and oceanic bird species, and recently demonstrated to impact some terrestrial songbirds to a comparable extent. It can bioaccumulate to concentrations of >1 µg/g in tissues of prey organisms such as fish and insects. At high enough concentrations, exposure to mercury is lethal to birds. However, environmental exposures are usually far below the lethal concentrations established by dosing studies.The objective of this review is to better understand the effects of sublethal exposure to mercury in birds. We restricted our survey of the literature to studies with at least some exposures >5 µg/g. The majority of sublethal effects were subtle and some studies of similar endpoints reached different conclusions. Strong support exists in the literature for the conclusion that mercury exposure reduces reproductive output, compromises immune function, and causes avoidance of high-energy behaviors. For some endpoints, notably certain measures of reproductive success, endocrine and neurological function, and body condition, there is weak or contradictory evidence of adverse effects and further study is required. There was no evidence that environmentally relevant mercury exposure affects longevity, but several of the sublethal effects identified likely do result in fitness reductions that could adversely impact populations. Overall, 72% of field studies and 91% of laboratory studies found evidence of deleterious effects of mercury on some endpoint, and thus we can conclude that mercury is harmful to birds, and the many effects on reproduction indicate that bird population declines may already be resulting from environmental mercury pollution.

Experimental Infection and Clearance of Coccidian Parasites in Mercury-Exposed Zebra Finches.

Mercury is a globally distributed, persistent environmental contaminant that affects the health of many taxa. It can suppress the immune system, which often plays a role in defense against parasites. However, there have been few investigations of whether mercury affects the abilities of animals to resist parasitic infection. Here, we exposed zebra finches to a lifetime dietary exposure of methylmercury (1.2 µg/g wet weight) and experimentally infected them with coccidian parasites to examine the effect of methylmercury exposure on parasitic infection. The mercury-exposed birds did not have an altered immune response (heterophil:lymphocyte ratio) nor a reduced ability to clear the infection. However, mercury-exposed birds tended to have higher parasite loads at the time when we expected the greatest immune response (2-3 weeks post-infection). Although mercury did not greatly influence the infection-course of this parasite in captivity, responses may be more accentuated in the wild where birds face additional immune challenges.

Mercury alters initiation and construction of nests by zebra finches, but not incubation or provisioning behaviors.

Mercury is an environmental contaminant that impairs avian reproduction, but the behavioral and physiological mechanisms underlying this effect are poorly understood. The objective of this study was to determine whether lifetime dietary exposure to mercury (1.2 µg/g wet weight in food) impacted avian parental behaviors, and how this might influence reproductive success. To distinguish between the direct effects of mercury on parents and offspring, we created four treatment groups of captive-bred zebra finches (Taeniopygia guttata), with control and mercury-exposed adults raising cross-fostered control or mercury-exposed eggs (from maternal transfer). Control parents were 23% more likely to fledge young than parents exposed to mercury, regardless of egg exposure. Mercury-exposed parents were less likely to initiate nests than controls and spent less time constructing them. Nests of mercury-exposed pairs were lighter, possibly due to an impaired ability to bring nest material into the nestbox. However, nest temperature, incubation behavior, and provisioning rate did not differ between parental treatments. Unexposed control eggs tended to have shorter incubation periods and higher hatching success than mercury-exposed eggs, but there was no effect of parental exposure on these parameters. We accidentally discovered that parent finches transfer some of their body burden of mercury to nestlings during feeding through secretion in the crop. These results suggest that, in mercury-exposed songbirds, pre-laying parental behaviors, combined with direct exposure of embryos to mercury, likely contribute to reduced reproductive success and should be considered in future studies. Further research is warranted in field settings, where parents are exposed to greater environmental challenges and subtle behavioral differences might have more serious consequences than were observed in captivity.

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.

No Effect of Lifelong Methylmercury Exposure on Oxidative Status in Zebra Finches (Taeniopygia guttata): A Demonstration of Methylmercury-Induced Selection?

Songbirds exposed to methylmercury (MeHg) often exhibit reduced reproductive success and cognitive abilities. To better understand whether oxidative stress plays a role, we dosed zebra finches (Taeniopygia guttata) with a contaminated (1.2 ppm MeHg-cysteine) or control diet for their entire lives, including during development in the egg. Levels of antioxidant enzymes [superoxide dismutase (SOD1 and SOD2)], oxidative damage (4-hydroxynonenal; 4-HNE), and antioxidant transcription factors [nuclear factor (erythroid-derived 2)-like 2; Nrf2] were measured in the liver and pectoralis muscle of adults. MeHg treatment did not affect levels of 4-HNE or liver SOD2 or Nrf2. Birds in the MeHg treatment differed significantly from controls in pectoralis SOD1 and Nrf2, and tended to differ in liver SOD1 and pectoralis SOD2; however, we detected no overall pattern of effect of MeHg on oxidative status in dosed finches. We suspect that this is a consequence of the differential survival of MeHg-tolerant birds.

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.

Songbirds as sentinels of mercury in terrestrial habitats of eastern North America.

Mercury (Hg) is a globally distributed environmental contaminant with a variety of deleterious effects in fish, wildlife, and humans. Breeding songbirds may be useful sentinels for Hg across diverse habitats because they can be effectively sampled, have well-defined and small territories, and can integrate pollutant exposure over time and space. We analyzed blood total Hg concentrations from 8,446 individuals of 102 species of songbirds, sampled on their breeding territories across 161 sites in eastern North America [geometric mean Hg concentration = 0.25 µg/g wet weight (ww), range <0.01-14.60 µg/g ww]. Our records span an important time period-the decade leading up to implementation of the USEPA Mercury and Air Toxics Standards, which will reduce Hg emissions from coal-fired power plants by over 90 %. Mixed-effects modeling indicated that habitat, foraging guild, and age were important predictors of blood Hg concentrations across species and sites. Blood Hg concentrations in adult invertebrate-eating songbirds were consistently higher in wetland habitats (freshwater or estuarine) than upland forests. Generally, adults exhibited higher blood Hg concentrations than juveniles within each habitat type. We used model results to examine species-specific differences in blood Hg concentrations during this time period, identifying potential Hg sentinels in each region and habitat type. Our results present the most comprehensive assessment of blood Hg concentrations in eastern songbirds to date, and thereby provide a valuable framework for designing and evaluating risk assessment schemes using sentinel songbird species in the time after implementation of the new atmospheric Hg standards.

Plumage color and reproductive output of eastern bluebirds (Sialia sialis) nesting near a mercury-contaminated river.

Despite the growing evidence of mercury's impact on ecosystems, few studies have looked at the environmental impact of mercury pollution on terrestrial songbirds and the complex ways through which mercury might influence their fitness. In 2007-2008 eastern bluebirds (Sialia sialis) were monitored on mercury contaminated and reference sites for blood and feather mercury, reproductive success and plumage coloration. Higher tissue mercury accumulation was associated with plumage that was overall brighter and shifted towards the UV portion of the spectrum. In females, long-term mercury exposure, as indicated by feather mercury, was associated with smaller clutches of eggs. In males, recent mercury exposure, as indicated by blood mercury, was associated with a reduction in the proportion of hatchlings that fledged, potentially through reduced male provisioning of offspring. Reproductive success and plumage color are closely linked in bluebirds through mate choice, and our findings indicate that mercury contamination is associated with reproductive success directly and possibly indirectly, through coloration of bluebirds.

Tissue Mercury Concentrations and Survival of Tree Swallow Embryos, Nestlings and Young Adult Females on a Contaminated Site.

Tree swallows nesting on mercury-contaminated sites along the South River in Virginia, USA were monitored for reproductive success. The bodies of nestlings found deceased in their nest boxes were collected, along with blood and feather samples from the adult parents and surviving siblings. We also measured hatching and fledging success of the clutches and the annual recapture rate of adults. We found that the body feathers of deceased nestlings contained significantly higher concentrations of mercury (12.89 ± 8.42 µg/g, n = 15) than those of nestlings that survived to fledge (7.41 ± 4.79 µg/g, n = 15). However, mothers of more successful clutches (>75 % hatching) did not differ in mercury concentrations from females with less successful clutches (<50 % hatching). Additionally, adult females breeding for the first time that returned to breed the following year did not differ in blood mercury from females of the same age that bred once but never returned. Our results suggest that mercury had its greatest effect on these songbirds during the nestling stage, whereas for embryos or first-time breeding females, other factors likely played larger roles in mortality.

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.

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.

The role of weather in mediating the effect of mercury exposure on reproductive success in tree swallows.

Mercury is a heavy metal that has contaminated countless ecosystems throughout the world. A large body of literature has documented reproductive, physiological, and behavioral impairments associated with mercury exposure in laboratory settings, but whether and how such effects are manifest in free-living populations remains poorly understood. The purpose of this study was to evaluate whether tree swallow (Tachycineta bicolor) breeding success at a site with high mercury exposure varied with ambient temperature or precipitation at various points in the breeding cycle. Tree swallows nesting along the South River had significantly elevated blood total mercury (mean ± SE: 3.03 ± 0.15 µg/g) compared to swallows breeding on reference sites (mean ± SE: 0.16 ± 0.005 µg/g). These high levels of mercury were associated with reduced hatching and fledging success, and contaminated birds produced approximately one less fledgling per nest than their reference counterparts. The magnitude of this difference was weather-dependent: unusually high ambient temperatures encountered early in the nestling period were associated with reduced reproductive output in contaminated, but not in reference, birds. In contrast, little effect of mercury on success of nestlings was observed when temperatures were cooler, and precipitation also had no detectable interaction with mercury. These results provide insight into mechanisms underlying reproductive effects of mercury. In addition, these findings underscore the importance of considering variable environmental conditions when assessing effects of contaminants on free-living wildlife. In particular, projections about the effects of global climate change on ecotoxicological impacts must take into account the kinds of weather-mediated effect demonstrated here.