Gene Expression Profiles in Two Razor Clam Populations: Discerning Drivers of Population Status.

With rapidly changing marine ecosystems, shifts in abundance and distribution are being documented for a variety of intertidal species. We examined two adjacent populations of Pacific razor clams (Siliqua patula) in lower Cook Inlet, Alaska. One population (east) supported a sport and personal use fishery, but this has been closed since 2015 due to declines in abundance, and the second population (west) continues to support commercial and sport fisheries. We used gene expression to investigate potential causes of the east side decline, comparing razor clam physiological responses between east and west Cook Inlet. The target gene profile used was developed for razor clam populations in Alaska based on physiological responses to environmental stressors. In this study, we identified no differences of gene expression between east and west populations, leading to two potential conclusions: (1) differences in factors capable of influencing physiology exist between the east and west and are sufficient to influence razor clam populations but are not detected by the genes in our panel, or (2) physiological processes do not account for the differences in abundance, and other factors such as predation or changes in habitat may be impacting the east Cook Inlet population.

Differential Progression of Lymphoma in Two Captive Steller's Eiders (Polysticta stelleri).

This report details 2 different presentations of lymphoma in captive Steller's eiders (Polysticta stelleri). A female Steller's eider, at least 10 years old, developed lameness and lethargy. A complete blood cell count (CBC) revealed a severely elevated total white blood cell (WBC) count with a lymphocytosis. A subsequent liver biopsy confirmed a diagnosis of lymphoma and lymphocytic leukemia. The female Steller's eider was euthanatized 11 days after presentation. On necropsy, neoplastic lymphoid infiltrates were present in multiple tissues, primarily in the skin and subcutis. The results of a CBC from an apparently healthy male Steller's eider, at least 14 years old, also indicated this bird had an increased WBC count with a lymphocytosis. Serum submitted for protein electrophoresis indicated that this bird had a monoclonal gammopathy. Lymphocytes isolated from the male Steller's eider had low levels of proliferation in response to mitogens. The bird survived nearly 2 years without treatment, and the WBC count continued to increase during this time. The male Steller's eider maintained good body condition and exhibited normal behavior throughout the course of the disease until a sudden decline just prior to death. Lymphoma was diagnosed based on histopathology results. Infiltrates of atypical lymphocytes were observed in the bone marrow, proventriculus, ventriculus, heart, and eyes.

Contemporary diets of walruses in Bristol Bay, Alaska suggest temporal variability in benthic community structure.

BACKGROUND: Pacific walruses (Odobenus rosmarus divergens) are a conspicuous and important component of the Bristol Bay ecosystem and human social systems, but very little is known about walrus ecology in this region, principally their feeding ecology. The present work provides contemporary data on the diets of walruses at four haulout locations throughout Bristol Bay between 2014 and 2018. METHODS: We analyzed scat and gastrointestinal tract samples from these animals using quantitative polymerase chain reaction to amplify prey DNA, which allowed for diet estimates based on frequencies of prey item occurrence and on the relative importance of dietary items as determined from DNA threshold cycle scores. RESULTS: Diets were highly diverse at all locations, but with some variation in composition that may be related to the time of year that samples were collected (summer vs. autumn), or to spatial variability in the distribution of prey. Overall, polychaetes and tunicates had the highest frequencies of occurrence and relative abundances in 2014-15, but a major change in diet appears to have occurred by 2017-18. While some sample sizes were small, diets in these later years contrasted sharply, with a greater prevalence of sea cucumbers and mollusks, and reduced importance of decapods and fishes compared to the earlier years. Prey identified in scat samples from one collection site also contrasted sharply with those reported from the same location in 1981. The apparent temporal shifts in walrus prey may represent a changing benthic ecosystem due to warming waters in recent decades.

Monitoring nearshore ecosystem health using Pacific razor clams (Siliqua patula) as an indicator species.

An emerging approach to ecosystem monitoring involves the use of physiological biomarker analyses in combination with gene transcription assays. For the first time, we employed these tools to evaluate the Pacific razor clam (Siliqua patula), which is important both economically and ecologically, as a bioindicator species in the northeast Pacific. Our objectives were to (1) develop biomarker and gene transcription assays with which to monitor the health of the Pacific razor clam, (2) acquire baseline biomarker and gene transcription reference ranges for razor clams, (3) assess the relationship between physiological and gene transcription assays and (4) determine if site-level differences were present. Pacific razor clams were collected in July 2015 and 2016 at three sites within each of two national parks in southcentral Alaska. In addition to determining reference ranges, we found differences in biomarker assay and gene transcription results between parks and sites which indicate variation in both large-scale and local environmental conditions. Our intent is to employ these methods to evaluate Pacific razor clams as a bioindicator of nearshore ecosystem health. Links between the results of the biomarker and gene transcription assays were observed that support the applicability of both assays in ecosystem monitoring. However, we recognize the need for controlled studies to examine the range of responses in physiology and gene transcripts to different stressors.

Physiological and gene transcription assays to assess responses of mussels to environmental changes.

Coastal regions worldwide face increasing management concerns due to natural and anthropogenic forces that have the potential to significantly degrade nearshore marine resources. The goal of our study was to develop and test a monitoring strategy for nearshore marine ecosystems in remote areas that are not readily accessible for sampling. Mussel species have been used extensively to assess ecosystem vulnerability to multiple, interacting stressors. We sampled bay mussels (Mytilus trossulus) in 2015 and 2016 from six intertidal sites in Lake Clark and Katmai National Parks and Preserves, in south-central Alaska. Reference ranges for physiological assays and gene transcription were determined for use in future assessment efforts. Both techniques identified differences among sites, suggesting influences of both large-scale and local environmental factors and underscoring the value of this combined approach to ecosystem health monitoring.

Immune parameters in different age classes of captive male Steller's eiders (Polysticta stelleri).

The immune system is important for host defense against antigens, but little is known about Steller's eider (Polysticta stelleri) immunology. This study compared hematological parameters, serum protein levels, lymphocyte proliferation, heat shock protein levels and oxidative damage in four different age classes of captive male Steller's eiders. The hatch year cohort had significantly higher total white blood cell and lymphocyte counts. The second year cohort had significantly higher albumin, alpha globulins and lymphocyte proliferation, and significantly lower beta globulin levels. The 9 year old males had a significantly higher IgY:IgY(ΔFc) ratio. The oldest eiders in the study, 14 + year old males, had significantly higher serum IgY, pre-albumin and glutathione reductase activity, and the lowest lymphocyte proliferation. This study provided a baseline of immune parameters in captive male Steller's eiders, and the results suggested the parameters were influenced by age-related changes.

Non-linear effect of sea ice: Spectacled Eider survival declines at both extremes of the ice spectrum.

Understanding the relationship between environmental factors and vital rates is an important step in predicting a species' response to environmental change. Species associated with sea ice are of particular concern because sea ice is projected to decrease rapidly in polar environments with continued levels of greenhouse gas emissions. The relationship between sea ice and the vital rates of the Spectacled Eider, a threatened species that breeds in Alaska and Russia and winters in the Bering Sea, appears to be complex. While severe ice can impede foraging for benthic prey, ice also suppresses wave action and provides a platform on which eiders roost, thereby reducing thermoregulation costs. We analyzed a 23-year mark-recapture dataset for Spectacled Eiders nesting on Kigigak Island in western Alaska, and tested survival models containing different ice and weather-related covariates. We found that much of the variation in eider survival could be explained by the number of days per year with >95% sea ice concentration at the Bering Sea core wintering area. Furthermore, the data supported a quadratic relationship with sea ice rather than a linear one, indicating that intermediate sea ice concentrations were optimal for survival. We then used matrix population models to project population trajectories using General Circulation Model (GCM) outputs of daily sea ice cover. GCMs projected reduced sea ice at the wintering area by year 2100 under a moderated emissions scenario (RCP 4.5) and nearly ice-free conditions under an unabated emissions scenario (RCP 8.5). Under RCP 4.5, stochastic models projected an increase in population size until 2069 coincident with moderate ice conditions, followed by a decline in population size as ice conditions shifted from intermediate to mostly ice-free. Under RCP 8.5, eider abundance increased until 2040 and then decreased to near extirpation toward the end of the century as the Bering Sea became ice-free. Considerable uncertainty around parameter estimates for survival in years with minimal sea ice contributed to variation in stochastic projections of future population size, and this uncertainty could be reduced with additional survival data from low-ice winters.

Monitoring for the Management of Disease Risk in Animal Translocation Programmes.

Monitoring is best viewed as a component of some larger programme focused on science or conservation. The value of monitoring is determined by the extent to which it informs the parent process. Animal translocation programmes are typically designed to augment or establish viable animal populations without changing the local community in any detrimental way. Such programmes seek to minimize disease risk to local wild animals, to translocated animals, and in some cases to humans. Disease monitoring can inform translocation decisions by (1) providing information for state-dependent decisions, (2) assessing progress towards programme objectives, and (3) permitting learning in order to make better decisions in the future. Here we discuss specific decisions that can be informed by both pre-release and post-release disease monitoring programmes. We specify state variables and vital rates needed to inform these decisions. We then discuss monitoring data and analytic methods that can be used to estimate these state variables and vital rates. Our discussion is necessarily general, but hopefully provides a basis for tailoring disease monitoring approaches to specific translocation programmes.

Phylogenetic and pathogenic characterization of novel adenoviruses isolated from long-tailed ducks (Clangula hyemalis).

Novel adenoviruses were isolated from a long-tailed duck (Clangula hyemalis) mortality event near Prudhoe Bay, Alaska in 2000. The long-tailed duck adenovirus genome was approximately 27 kb. A 907 bp hexon gene segment was used to design primers specific for the long-tailed duck adenovirus. Nineteen isolates were phylogenetically characterized based on portions of their hexon gene and 12 were most closely related to Goose adenovirus A. The remaining 7 shared no hexon sequences with any known adenoviruses. Experimental infections of mallards with a long-tailed duck reference adenovirus caused mild lymphoid infiltration of the intestine and paint brush hemorrhages of the mucosa and dilation of the intestine. This study shows novel adenoviruses from long-tailed ducks are diverse and provides further evidence that they should be considered in cases of morbidity and mortality in sea ducks. Conserved and specific primers have been developed that will help screen sea ducks for adenoviral infections.

The influence of year, laying date, egg fertility and incubation, individual hen, hen age and mass and clutch size on maternal immunoglobulin Y concentration in captive Steller's and spectacled eider egg yolk.

Steller's eiders and spectacled eiders are sea duck species whose populations have declined significantly and infectious diseases could influence offspring survival. Therefore, the maternal transfer of immunoglobulin Y (IgY) into yolk was investigated in captive Steller's and spectacled eiders during the 2007-2013 breeding seasons. This project had two objectives: establish baseline IgY levels in Steller's and spectacled eider yolk under controlled captive conditions and evaluate the effect of year, laying date, egg fertility, egg incubation duration, individual hen, hen age and mass, and laying order to determine which variables influenced IgY levels. Average IgY concentrations were 0.03-0.48 mg ml(-1) in Steller's eider yolk and 0.10-0.51 mg ml(-1) in spectacled eider yolk. The year and individual hen influenced IgY concentration in Steller's and spectacled eider yolk. The laying date was negatively correlated with egg IgY levels for most Steller's eider hens, but laying order was positively correlated with egg IgY concentration for spectacled eiders.

Longitudinal monitoring of sex steroid hormones in excrement of spectacled eiders (Somateria fischeri).

From the 1970s to the 1990s, the breeding population of spectacled eiders (Somateria fischeri) in western Alaska declined by 96%, which led to the listing of this species as threatened under the Endangered Species Act in 1993. Since then, the population has stabilized, but has not recovered to pre-decline numbers. While little is known about reproductive endocrinology in spectacled eiders, in other avian species, estrogen and testosterone are known to initiate and modulate various reproductive processes including yolk protein synthesis, reproductive behaviors and secondary sex characteristics. Measurement of the metabolites of estrogen and testosterone (EM and TM, respectively) in excrement reflect circulating hormone concentrations and provide a non-invasive method to monitor reproductive physiology. We measured concentrations of excreted EM in captive females and TM in males to (1) determine the efficacy of commercially available radioimmunoassay kits to detect EM and TM, (2) describe annual profiles of EM and TM concentrations, and (3) define the reproductive season of captive spectacled eiders using endocrine status. Excrement samples were collected from captive female and male spectacled eiders three times per week throughout 1 year. Female EM and male TM levels were quantified using radioimmunoassay. Mean female EM profile exhibited values exceeding the threshold for "peak" values (EM>193.3 ng/g) from mid-February to early July, and again in September. Additionally, the highest average concentrations of EM were seen in March, May and September. Elevated TM concentrations occurred in mid March, mid May and late June. These data suggest that levels of excreted sex steroids reflect patterns predicted by breeding landmarks in the annual cycle and will assist in field monitoring and captive breeding programs for spectacled eiders.

Context dependency of baseline glucocorticoids as indicators of individual quality in a capital breeder.

Identifying markers of individual quality is a central goal of life-history theory and conservation biology. The 'corticosterone (CORT)-fitness hypothesis' postulates that low fitness signals impaired ability to cope with the environment, resulting in elevated baseline CORT levels. CORT can, however, be negatively, positively or neutrally related to fitness, depending on the context. In order to clarify this controversial issue, we elucidate the utility of using baseline CORT as a correlate of individual fitness in incubating female eiders across variable environments. An increase in serum CORT with decreasing body condition was evident in older, more experienced breeders, while increased clutch mass was associated with elevated serum CORT in females breeding late in the season. For faecal CORT, the expected negative association with body condition was observed only in early breeders. We found a strong increase in faecal CORT with increasing baseline body temperature, indicating the utility of body temperature as a complementary stress indicator. Females in good body condition had a lower baseline body temperature, but this effect was only observed on open islands, a harsher breeding habitat less buffered against weather variability. Females with higher reproductive investment also maintained a lower baseline body temperature. Nest success strongly decreased with increasing serum and faecal CORT concentrations, and individual stress hormone and body temperature profiles were repeatable over years. Although our data support the tenet that baseline CORT is negatively related to fitness, the complex context-dependent effects call for cautious interpretation of relationships between stress physiology and phenotypic quality.

Carbon isotopic fractionation in eider adipose tissue varies with fatty acid structure: implications for trophic studies.

Carbon isotopic fractionation was investigated in fatty acids (FA) of adipose tissue and blood serum of threatened Steller's eiders (Polysticta stelleri) and spectacled eiders (Somateria fischeri) relative to the FA in their diets. Captive eiders were fed a known diet for 180 days with serum sampled at 60, 120 and 180 days immediately after a 12 fast; adipose was collected at 180 days. Essential FA (EFA) in the adipose showed varying degrees of isotope fractionation (0-4‰), depending on FA structure. The δ(13)C values of long-chain FA 20:5n-3 and 22:6n-3 did not differ from those in the diet, while those of 18:2n-6 and 18:3n-3 were ∼2‰ greater than in the diet. The δ(13)C values of free FA (FFA) in serum were not consistent within individuals or sampling dates; fractionation varied randomly, suggesting that FFA were arising from diet, rather than mobilization from adipose tissue. Discrimination factors were used in combination with a mixing model incorporating FA and lipid concentrations to estimate the diet of eiders fed a binary mixture with contrasting isotopic signatures. Diet estimates varied with FA but mean values closely approximated the actual proportions consumed. By tracking EFA, this study avoided the complications in interpretation arising from isotopic routing of carbon in bulk isotope analyses and serves as a basis for the development of compound-specific isotopic methods to trace dietary input in wild eiders. However, our understanding of the processes contributing to the variation in isotopic signatures of FA in nature is currently limited, and we recommend that future research directions focus on elucidating these mechanisms.

Molecular typing of Escherichia coli strains associated with threatened sea ducks and near-shore marine habitats of south-west Alaska.

In Alaska, sea ducks winter in coastal habitats at remote, non-industrialized areas, as well as in proximity to human communities and industrial activity. We evaluated prevalence and characteristics of Escherichia coli strains in faecal samples of Steller's eiders (Polysticta stelleri; n = 122) and harlequin ducks (Histrionicus histrionicus; n = 21) at an industrialized site and Steller's eiders (n = 48) at a reference site, and compared these strains with those isolated from water samples from near-shore habitats of ducks. The overall prevalence of E. coli was 16% and 67% in Steller's eiders and harlequin ducks, respectively, at the industrialized study site, and 2% in Steller's eiders at the reference site. Based on O and H antigen subtyping and genetic characterization by enterobacterial repetitive intergenic consensus polymerase chain reaction and pulsed-field gel electrophoresis, we found evidence of avian pathogenic E. coli (APEC) strains associated with both species and detected E. coli strains carrying virulence genes associated with mammals in harlequin ducks. Steller's eiders that carried APEC had lower serum total protein and albumin concentrations, providing further evidence of pathogenicity. The genetic profile of two E. coli strains from water matched an isolate from a Steller's eider providing evidence of transmission between near-shore habitats and birds.

Validating quantitative fatty acid signature analysis to estimate diets of spectacled and Steller's eiders (Somateria fischeri and Polysticta stelleri).

Fatty acid (FA) signature analysis has been used to study foraging ecology and food webs in marine ecosystems. This powerful method provides information about diets over an extended time period (e.g., 2-4 weeks), rather than just the most recent meal as with most traditional approaches. Using consumer FA signatures, along with a comprehensive database of diet FA signatures, and accounting for consumer FA metabolism, it is possible to estimate the proportions of diet items in the consumer's diet using quantitative FA signature analysis (QFASA). However, before applying QFASA to free-ranging populations, ideally, controlled feeding studies are performed to determine FA deposition and turnover characteristics. We conducted feeding experiments to validate QFASA in captive spectacled eiders (Somateria fischeri) and Steller's eiders (Polysticta stelleri) as a minimally invasive method for studying the diets of these threatened species. We determined FA deposition in eider adipose tissue relative to long-term diet, and developed calibration coefficients (CCs) to account for eider lipid metabolism. Using these CCs with subsequent diet trials, QFASA accurately indicated diet and diet switches. QFASA estimates also indicated that turnover of dietary FAs was not complete by 21 or 29 days, and confirmed that diets could be estimated over an extended period of >29 days. Thus, our understanding of diet can be backtracked to more than a month in captive feeding eiders. We conclude that applying QFASA techniques to eiders and other birds in the wild has the potential to provide valuable information about their diets at various life history stages.

Effects of ACTH, capture, and short term confinement on glucocorticoid concentrations in harlequin ducks (Histrionicus histrionicus).

Little is known about baseline concentrations of adrenal hormones and hormonal responses to stress in sea ducks, although significant population declines documented in several species suggest that sea ducks are exposed to increased levels of environmental stress. Such declines have been observed in geographically distinct harlequin duck populations. We performed an adrenocorticotropic hormone (ACTH) challenge to evaluate adrenal function and characterize corticosterone concentrations in captive harlequin ducks and investigated the effects of capture, surgery, and short term confinement on corticosterone concentrations in wild harlequin ducks. Harlequin ducks responded to the ACTH challenge with an average three-fold increase in serum corticosterone concentration approximately 90 min post injection, and a four- to five-fold increase in fecal glucocorticoid concentration 2 to 4 h post injection. Serum corticosterone concentrations in wild harlequin ducks increased within min of capture and elevated levels were found for several hours post capture, indicating that surgery and confinement maintain elevated corticosterone concentrations in this species. Mean corticosterone concentrations in wild harlequin ducks held in temporary captivity were similar to the maximum response levels during the ACTH challenge in captive birds. However, large variation among individuals was observed in responses of wild birds, and we found additional evidence suggesting that corticosterone responses varied between hatch year and after hatch year birds.

Polycyclic aromatic hydrocarbon exposure in Steller's eiders (Polysticta stelleri) and Harlequin ducks (Histronicus histronicus) in the eastern Aleutian Islands, Alaska, USA.

Seaducks may be affected by harmful levels of polycyclic aromatic hydrocarbons (PAHs) at seaports near the Arctic. As an indicator of exposure to PAHs, we measured hepatic enzyme 7-ethoxyresorufin-O-deethylase activity (EROD) to determine cytochrome P4501A induction in Steller's eiders (Polysticta stelleri) and Harlequin ducks (Histronicus histronicus) from Unalaska, Popof, and Unga Islands (AK, USA) in 2002 and 2003. We measured PAHs and organic contaminants in seaduck prey samples and polychlorinated biphenyl congeners in seaduck blood plasma to determine any relationship to EROD. Using Akaike's information criterion, species and site differences best explained EROD patterns: Activity was higher in Harlequin ducks than in Steller's eiders and higher at industrial than at nonindustrial sites. Site-specific concentrations of PAHs in blue mussels ([Mytilus trossilus] seaduck prey; PAH concentrations higher at Dutch Harbor, Unalaska, than at other sites) also was important in defining EROD patterns. Organochlorine compounds rarely were detected in prey samples. No relationship was found between polychlorinated biphenyl congeners in avian blood and EROD, which further supported inferences derived from Akaike's information criterion. Congeners were highest in seaducks from a nonindustrial or reference site, contrary to PAH patterns. To assist in interpreting the field study, 15 captive Steller's eiders were dosed with a PAH known to induce cytochrome P4501A. Dosed, captive Steller's eiders had definitive induction, but results indicated that wild Steller's eiders were exposed to PAHs or other inducing compounds at levels greater than those used in laboratory studies. Concentrations of PAHs in blue mussels at or near Dutch Harbor (approximately 1,180-5,980 ng/g) approached those found at highly contaminated sites (approximately 4,100-7,500 ng/g).

Effects of dietary selenium on tissue concentrations, pathology, oxidative stress, and immune function in common eiders (Somateria mollissima).

Common eiders (Somateria mollissima) were fed added Se (as L-selenomethionine) in concentrations increasing from 10 to 80 ppm in a pilot study (Study 1) or 20 (low exposure) and up to 60 (high exposure) ppm Se in Study 2. Body weights of Study 1 ducks and high-exposure ducks in Study 2 declined rapidly. Mean concentrations of Se in blood reached 32.4 ppm wet weight in Study 1 and 17.5 ppm wet weight in high-exposure birds in Study 2. Mean Se concentrations in liver ranged from 351 (low exposure, Study 2) to 1252 ppm dry weight (Study 1). Oxidative stress was evidenced by Se-associated effects on glutathione metabolism. As Se concentrations in liver increased, Se-dependent glutathione peroxidase activity, glutathione reductase activity, oxidized glutathione levels, and the ratio of hepatic oxidized to reduced glutathione increased. In Study 2, the T-cell-mediated immune response was adversely affected in high-exposure eiders, but ducks in the low-exposure group exhibited evidence of an enhanced antibody-mediated immune response. Gross lesions in high-exposure ducks included emaciation, absence of thymus, and loss of nails from digits. Histologic lesions included severe depletion of lymphoid organs, hepatopathy, and necrosis of feather pulp and feather epithelium. Field studies showed that apparently healthy sea ducks generally have higher levels of Se in liver than healthy fresh-water birds, but lower than concentrations found in our study. Data indicate that common eiders and probably other sea ducks possess a higher threshold, or adverse effect level, for Se in tissues than fresh-water species. However, common eiders developed signs of Se toxicity similar to those seen in fresh-water birds.

Eider females form non-kin brood-rearing coalitions.

Kin selection is a powerful tool for understanding cooperation among individuals, yet its role as the sole explanation of cooperative societies has recently been challenged on empirical grounds. These studies suggest that direct benefits of cooperation are often overlooked, and that partner choice may be a widespread mechanism of cooperation. Female eider ducks (Somateria mollissima) may rear broods alone, or they may pool their broods and share brood-rearing. Females are philopatric, and it has been suggested that colonies may largely consist of related females, which could promote interactions among relatives. Alternatively, shared brood care could be random with respect to relatedness, either because brood amalgamations are accidental and nonadaptive, or through group augmentation, assuming that the fitness of all group members increases with group size. We tested these alternatives by measuring the relatedness of co-tending eider females in enduring coalitions with microsatellite markers. Females formed enduring brood-rearing coalitions with each other at random with respect to relatedness. However, based on previous data, partner choice is nonrandom and dependent on female body condition. We discuss potential mechanisms underlying eider communal brood-rearing decisions, which may be driven by the specific ecological conditions under which sociality has evolved in this species.

Evidence of chromosomal damage in common eiders (Somateria mollissima) from the Baltic Sea.

Common eiders nesting in the Baltic Sea are exposed to generally high levels of contaminants including potentially genotoxic polycyclic aromatic hydrocarbons and organochlorines. Blood samples were collected from eiders at eight sites in the Baltic Sea and two sites in the Beaufort Sea. DNA content variation was estimated using the flow cytometric method, and subsequently utilized as a biomarker of genetic damage. We observed no significant differences in genetic damage among populations within either the Baltic or Beaufort Seas. However, eider populations from the Baltic Sea had significantly elevated estimates of genetic damage compared to populations from the Beaufort Sea.