Reference Intervals for Serum Biochemistries of Molting Pacific Black Brant ( Branta bernicla nigricans) in Northern Alaska, USA.

We determined reference intervals for nine serum biochemistries in samples from 329 molting, after-hatch-year, Pacific Black Brant ( Branta bernicla nigricans) in Alaska, US. Cholesterol and nonesterified fatty acids differed by sex, but no other differences were noted.

Blood selenium concentrations in female Pacific black brant molting in Arctic Alaska: Relationships with age and habitat salinity.

Blood samples collected from 81 female Pacific black brant (Branta bernicla nigricans) molting near Teshekpuk Lake, Alaska, were analyzed for selenium concentration. The concentration of selenium in blood of after second year (hatched two or more years ago) females (0.84µg/g wet weight) was significantly greater than the concentration in second year (hatched the previous year) females (0.61µg/g wet weight). The concentrations of selenium we found in blood of black brant were 1.5 to 2 times greater than baseline values typical of freshwater birds, but considerably lower than reported in other marine waterfowl sampled in Alaska. This finding may be attributable in part to the nearly exclusive herbivorous diet of black brant. No relationship was noted between blood selenium concentration and molting habitat salinity. We are unaware of any previous reports of blood selenium concentrations in black brant.

Wild bird surveillance for highly pathogenic avian influenza H5 in North America.

It is unknown how the current Asian origin highly pathogenic avian influenza H5 viruses arrived, but these viruses are now poised to become endemic in North America. Wild birds harbor these viruses and have dispersed them at regional scales. What is unclear is how the viruses may be moving from the wild bird reservoir into poultry holdings. Active surveillance of live wild birds is likely the best way to determine the true distribution of these viruses. We also suggest that sampling be focused on regions with the greatest risk for poultry losses and attempt to define the mechanisms of transfer to enhance biosecurity. Responding to the recent outbreaks of highly pathogenic avian influenza in North America requires an efficient plan with clear objectives and potential management outcomes.

Genomic analysis of avian influenza viruses from waterfowl in western Alaska, USA.

The Yukon-Kuskokwim Delta (Y-K Delta) in western Alaska is an immense and important breeding ground for waterfowl. Migratory birds from the Pacific Americas, Central Pacific, and East Asian-Australasian flyways converge in this region, providing opportunities for intermixing of North American- and Eurasian-origin hosts and infectious agents, such as avian influenza virus (AIV). We characterized the genomes of 90 low pathogenic (LP) AIV isolates from 11 species of waterfowl sampled on the Y-K Delta between 2006 and 2009 as part of an interagency surveillance program for the detection of the H5N1 highly pathogenic (HP) strain of AIV. We found evidence for subtype and genetic differences between viruses from swans and geese, dabbling ducks, and sea ducks. At least one gene segment in 39% of all isolates was Eurasian in origin. Target species (those ranked as having a relatively high potential to introduce HP H5N1 AIV to North America) were no more likely than nontarget species to carry viruses with genes of Eurasian origin. These findings provide evidence that the frequency at which viral gene segments of Eurasian origin are detected does not result from a strong species effect, but rather we suspect it is linked to the geographic location of the Y-K Delta in western Alaska where flyways from different continents overlap. This study provides support for retaining the Y-K Delta as a high priority region for the surveillance of Asian avian pathogens such as HP H5N1 AIV.

High seroprevalence of antibodies to avian influenza viruses among wild waterfowl in Alaska: implications for surveillance.

We examined seroprevalence (presence of detectable antibodies in serum) for avian influenza viruses (AIV) among 4,485 birds, from 11 species of wild waterfowl in Alaska (1998-2010), sampled during breeding/molting periods. Seroprevalence varied among species (highest in eiders (Somateria and Polysticta species), and emperor geese (Chen canagica)), ages (adults higher than juveniles), across geographic locations (highest in the Arctic and Alaska Peninsula) and among years in tundra swans (Cygnus columbianus). All seroprevalence rates in excess of 60% were found in marine-dependent species. Seroprevalence was much higher than AIV infection based on rRT-PCR or virus isolation alone. Because pre-existing AIV antibodies can infer some protection against highly pathogenic AIV (HPAI H5N1), our results imply that some wild waterfowl in Alaska could be protected from lethal HPAIV infections. Seroprevalence should be considered in deciphering patterns of exposure, differential infection, and rates of AIV transmission. Our results suggest surveillance programs include species and populations with high AIV seroprevalences, in addition to those with high infection rates. Serologic testing, including examination of serotype-specific antibodies throughout the annual cycle, would help to better assess spatial and temporal patterns of AIV transmission and overall disease dynamics.

Chronic hydrocarbon exposure of harlequin ducks in areas affected by the Selendang Ayu oil spill at Unalaska Island, Alaska.

We evaluated chronic exposure of harlequin ducks (Histrionicus histrionicus) to hydrocarbons associated with the 2004 M/V Selendang Ayu oil spill at Unalaska Island, Alaska. We measured levels of hepatic 7-ethoxyresorufin-O-deethylase activity (EROD) in liver biopsy samples as an indicator of hydrocarbon exposure in three oiled bays and one reference bay in 2005, 2006, and 2008. Median EROD activity in ducks from oiled bays was significantly higher than in the reference bay in seven of nine pairwise comparisons. These results indicated that harlequin ducks were exposed to lingering hydrocarbons more than three years after the spill.

Selenium concentrations and enzyme activities of glutathione metabolism in wild long-tailed ducks and common eiders.

The relationships of selenium (Se) concentrations in whole blood with plasma activities of total glutathione peroxidase, Se-dependent glutathione peroxidase, and glutathione reductase were studied in long-tailed ducks (Clangula hyemalis) and common eiders (Somateria mollissima) sampled along the Beaufort Sea coast of Alaska, USA. Blood Se concentrations were >8 µg/g wet weight in both species. Linear regression revealed that the activities of total and Se-dependent glutathione peroxidase were significantly related to Se concentrations only in long-tailed ducks, raising the possibility that these birds were experiencing early oxidative stress.

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.

Intercontinental reassortment and genomic variation of low pathogenic avian influenza viruses isolated from northern pintails (Anas acuta) in Alaska: examining the evidence through space and time.

Migration and population genetic data for northern pintails (Anas acuta) and phylogenetic analysis of low pathogenic avian influenza (LPAI) viruses from this host in Alaska suggest that northern pintails are involved in ongoing intercontinental transmission of avian influenza. Here, we further refine this conclusion through phylogenetic analyses which demonstrate that detection of foreign lineage gene segments is spatially dependent and consistent through time. Our results show detection of foreign lineage gene segments to be most likely at sample locations on the Alaska Peninsula and least likely along the Southern Alaska Coast. Asian lineages detected at four gene segments persisted across years, suggesting maintenance in avian hosts that migrate to Alaska each year from Asia or in hosts that remain in Alaska throughout the year. Alternatively, live viruses may persist in the environment and re-infect birds in subsequent seasons.

Relationships between hepatic trace element concentrations, reproductive status, and body condition of female greater scaup.

We collected female greater scaup (Aythya marila) on the Yukon-Kuskokwim Delta, Alaska during two breeding seasons to determine if concentrations of 18 trace elements in livers and eggs were elevated and if hepatic concentrations correlated with body condition or affected reproductive status. Fifty-six percent, 5%, and 42% of females, respectively, had elevated hepatic cadmium (Cd: >3 microg g(-1) dry weight [dw]), mercury (Hg: >3 microg g(-1) dw), and selenium (Se: >10 microg g(-1) dw). Somatic protein and lipid reserves were not correlated with hepatic Cd or Hg, but there was a weak negative correlation between protein and Se. Hepatic Cd, Hg, and Se were similar in females that had and had not initiated egg production. In a sample of six eggs, 33% and 100%, respectively, contained Se and Hg, but concentrations were below embryotoxicity thresholds. We conclude that trace element concentrations documented likely were not adversely impacting this study population.

Avian influenza at both ends of a migratory flyway: characterizing viral genomic diversity to optimize surveillance plans for North America.

Although continental populations of avian influenza viruses are genetically distinct, transcontinental reassortment in low pathogenic avian influenza (LPAI) viruses has been detected in migratory birds. Thus, genomic analyses of LPAI viruses could serve as an approach to prioritize species and regions targeted by North American surveillance activities for foreign origin highly pathogenic avian influenza (HPAI). To assess the applicability of this approach, we conducted a phylogenetic and population genetic analysis of 68 viral genomes isolated from the northern pintail (Anas acuta) at opposite ends of the Pacific migratory flyway in North America. We found limited evidence for Asian LPAI lineages on wintering areas used by northern pintails in California in contrast to a higher frequency on breeding locales of Alaska. Our results indicate that the number of Asian LPAI lineages observed in Alaskan northern pintails, and the nucleotide composition of LPAI lineages, is not maintained through fall migration. Accordingly, our data indicate that surveillance of Pacific Flyway northern pintails to detect foreign avian influenza viruses would be most effective in Alaska. North American surveillance plans could be optimized through an analysis of LPAI genomics from species that demonstrate evolutionary linkages with European or Asian lineages and in regions that have overlapping migratory flyways with areas of HPAI outbreaks.

Prevalence of Influenza A viruses in wild migratory birds in Alaska: patterns of variation in detection at a crossroads of intercontinental flyways.

BACKGROUND: The global spread of the highly pathogenic avian influenza H5N1 virus has stimulated interest in a better understanding of the mechanisms of H5N1 dispersal, including the potential role of migratory birds as carriers. Although wild birds have been found dead during H5N1 outbreaks, evidence suggests that others have survived natural infections, and recent studies have shown several species of ducks capable of surviving experimental inoculations of H5N1 and shedding virus. To investigate the possibility of migratory birds as a means of H5N1 dispersal into North America, we monitored for the virus in a surveillance program based on the risk that wild birds may carry the virus from Asia. RESULTS: Of 16,797 birds sampled in Alaska between May 2006 and March 2007, low pathogenic avian influenza viruses were detected in 1.7% by rRT-PCR but no highly pathogenic viruses were found. Our data suggest that prevalence varied among sampling locations, species (highest in waterfowl, lowest in passerines), ages (juveniles higher than adults), sexes (males higher than females), date (highest in autumn), and analytical technique (rRT-PCR prevalence = 1.7%; virus isolation prevalence = 1.5%). CONCLUSION: The prevalence of low pathogenic avian influenza viruses isolated from wild birds depends on biological, temporal, and geographical factors, as well as testing methods. Future studies should control for, or sample across, these sources of variation to allow direct comparison of prevalence rates.

Genetic evidence of intercontinental movement of avian influenza in a migratory bird: the northern pintail (Anas acuta).

The role of migratory birds in the movement of the highly pathogenic (HP) avian influenza H5N1 remains a subject of debate. Testing hypotheses regarding intercontinental movement of low pathogenic avian influenza (LPAI) viruses will help evaluate the potential that wild birds could carry Asian-origin strains of HP avian influenza to North America during migration. Previous North American assessments of LPAI genetic variation have found few Asian reassortment events. Here, we present results from whole-genome analyses of LPAI isolates collected in Alaska from the northern pintail (Anas acuta), a species that migrates between North America and Asia. Phylogenetic analyses confirmed the genetic divergence between Asian and North American strains of LPAI, but also suggested inter-continental virus exchange and at a higher frequency than previously documented. In 38 isolates from Alaska, nearly half (44.7%) had at least one gene segment more closely related to Asian than to North American strains of LPAI. Additionally, sequences of several Asian LPAI isolates from GenBank clustered more closely with North American northern pintail isolates than with other Asian origin viruses. Our data support the role of wild birds in the intercontinental transfer of influenza viruses, and reveal a higher degree of transfer in Alaska than elsewhere in North America.

Applying the scientific method when assessing the influence of migratory birds on the dispersal of H5N1.

BACKGROUND: The role of wild birds in the dispersal of highly pathogenic avian influenza virus H5N1 continues to be the subject of considerable debate. However, some researchers functionally examining the same question are applying opposing null hypotheses when examining this issue. DISCUSSION: I describe the correct method for establishing a null hypothesis under the scientific method. I suggest that the correct null hypothesis is that migratory birds can disperse this virus during migration and encourage researchers to design studies to falsify this null. Finally, I provide several examples where statements made during this debate, while strictly true, are not generally informative or are speculative. SUMMARY: By adhering to the scientific method, definitive answers regarding the role of wild birds in the dispersal of highly pathogenic viruses will be reached more effectively.

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).

Coupling contaminants with demography: effects of lead and selenium in Pacific common eiders.

We coupled intensive population monitoring with collection of blood samples from 383 nesting Pacific common eiders (Somateria mollisima v-nigrum) at two locations in Alaska (USA) from 2002 to 2004. We investigated annual, geographic, and within-season variation in blood concentrations of lead and selenium; compared exposure patterns with sympatrically nesting spectacled eiders (Somateria fischeri); and examined relationships with clutch size, egg viability, probability of hatching, and apparent survival of adult females. Lead concentrations were elevated in 3.6% of females, and all individuals exhibited elevated selenium, most (81%) at concentrations associated with death in captive waterfowl. Blood lead and selenium concentrations varied both within and among site-years and were lower than those of spectacled eiders. During incubation, blood lead concentrations in females increased significantly (possibly via re-release of stored lead from bone), whereas selenium concentrations decreased (likely because of natural excretion). Probability of a nest containing at least one nonviable egg was positively related to blood selenium in hens, but adverse effects in other life-history variables were not supported. Although reproduction appeared to be sensitive to selenium toxicity, our data suggest that high rates of nonviability are unlikely in this population and that selenium-related reductions to clutch size would be inconsequential at the scale of overall population dynamics. We conclude that Pacific common eiders and other wild marine birds likely have higher selenium tolerances than freshwater species and that interspecific differences in exposure levels may reflect differences in reproductive strategies.

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.

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.

An adenovirus linked to mortality and disease in long-tailed ducks (Clangula hyemalis) in Alaska.

An adenovirus was isolated from intestinal samples of two long-tailed ducks (Clangula hyemalis) collected during a die-off in the Beaufort Sea off the north coast of Alaska in 2000. The virus was not neutralized by reference antiserum against known group I, II, or III avian adenoviruses and may represent a new serotype. The prevalence of the virus was determined in live-trapped long-tailed ducks at the mortality site and at a reference site 100 km away where no mortality was observed. Prevalence of adenovirus antibodies in serum samples at the mortality site was 86% compared to 10% at the reference site. Furthermore, 50% of cloacal swabs collected at the mortality site and only 7% of swabs from the reference site were positive for adenoviruses. In 2001, no mortality was observed at either of the study areas, and virus prevalence in both serum and cloacal samples was low, providing further evidence that the adenovirus was linked to the mortality event in 2000. The virus was used to infect long-tailed ducks under experimental conditions and resulted in lesions previously described for avian adenovirus infections and similar to those observed in long-tailed duck carcasses from the Beaufort Sea. The status of long-tailed ducks has recently become a concern in Alaska due to precipitous declines in breeding populations there since the mid-1970s. Our findings suggest that the newly isolated adenovirus is a disease agent and source of mortality in long-tailed ducks, and thus could be a contributing factor in population declines.