Maintenance and dissemination of avian-origin influenza A virus within the northern Atlantic Flyway of North America.

Wild waterbirds, the natural reservoirs for avian influenza viruses, undergo migratory movements each year, connecting breeding and wintering grounds within broad corridors known as flyways. In a continental or global view, the study of virus movements within and across flyways is important to understanding virus diversity, evolution, and movement. From 2015 to 2017, we sampled waterfowl from breeding (Maine) and wintering (Maryland) areas within the Atlantic Flyway (AF) along the east coast of North America to investigate the spatio-temporal trends in persistence and spread of influenza A viruses (IAV). We isolated 109 IAVs from 1,821 cloacal / oropharyngeal samples targeting mallards (Anas platyrhynchos) and American black ducks (Anas rubripes), two species having ecological and conservation importance in the flyway that are also host reservoirs of IAV. Isolates with >99% nucleotide similarity at all gene segments were found between eight pairs of birds in the northern site across years, indicating some degree of stability among genome constellations and the possibility of environmental persistence. No movement of whole genome constellations were identified between the two parts of the flyway, however, virus gene flow between the northern and southern study locations was evident. Examination of banding records indicate direct migratory waterfowl movements between the two locations within an annual season, providing a mechanism for the inferred viral gene flow. Bayesian phylogenetic analyses provided evidence for virus dissemination from other North American wild birds to AF dabbling ducks (Anatinae), shorebirds (Charidriformes), and poultry (Galliformes). Evidence was found for virus dissemination from shorebirds to gulls (Laridae), and dabbling ducks to shorebirds and poultry. The findings from this study contribute to the understanding of IAV ecology in waterfowl within the AF.

Review of harmful algal bloom effects on birds with implications for avian wildlife in the Chesapeake Bay region.

The Chesapeake Bay, along the mid-Atlantic coast of North America, is the largest estuary in the United States and provides critical habitat for wildlife. In contrast to point and non-point source release of pesticides, metals, and industrial, personal care and household use chemicals on biota in this watershed, there has only been scant attention to potential exposure and effects of algal toxins on wildlife in the Chesapeake Bay region. As background, we first review the scientific literature on algal toxins and harmful algal bloom (HAB) events in various regions of the world that principally affected birds, and to a lesser degree other wildlife. To examine the situation for the Chesapeake, we compiled information from government reports and databases summarizing wildlife mortality events for 2000 through 2020 that were associated with potentially toxic algae and HAB events. Summary findings indicate that there have been few wildlife mortality incidents definitively linked to HABs, other mortality events that were suspected to be related to HABs, and more instances in which HABs may have indirectly contributed to or occurred coincident with wildlife mortality. The dominant toxins found in the Chesapeake Bay drainage that could potentially affect wildlife are microcystins, with concentrations in water approaching or exceeding human-based thresholds for ceasing recreational use and drinking water at a number of locations. As an increasing trend in HAB events in the U.S. and in the Chesapeake Bay have been reported, additional information on HAB toxin exposure routes, comparative sensitivity among species, consequences of sublethal exposure, and better diagnostic and risk criteria would greatly assist in predicting algal toxin hazard and risks to wildlife.

A Video Surveillance System to Monitor Breeding Colonies of Common Terns (Sterna Hirundo).

Many waterbird populations have faced declines over the last century, including the common tern (Sterna hirundo), a waterbird species with a widespread breeding distribution, that has been recently listed as endangered in some habitats of its range. Waterbird monitoring programs exist to track populations through time; however, some of the more intensive approaches require entering colonies and can be disruptive to nesting populations. This paper describes a protocol that utilizes a minimally invasive surveillance system to continuously monitor common tern nesting behavior in typical ground-nesting colonies. The video monitoring system utilizes wireless cameras focused on individual nests as well as over the colony as a whole, and allows for observation without entering the colony. The video system is powered with several 12 V car batteries that are continuously recharged using solar panels. Footage is recorded using a digital video recorder (DVR) connected to a hard drive, which can be replaced when full. The DVR may be placed outside of the colony to reduce disturbance. In this study, 3,624 h of footage recorded over 63 days in weather conditions ranging from 12.8 °C to 35.0 °C produced 3,006 h (83%) of usable behavioral data. The types of data retrieved from the recorded video can vary; we used it to detect external disturbances and measure nesting behavior during incubation. Although the protocol detailed here was designed for ground-nesting waterbirds, the principal system could easily be modified to accommodate alternative scenarios, such as colonial arboreal nesting species, making it widely applicable to a variety of research needs.

Analysis of free and metabolized microcystins in samples following a bird mortality event.

In the summer of 2012, over 750 dead and dying birds were observed at the Paul S. Sarbanes Ecosystem Restoration Project at Poplar Island, Maryland, USA (Chesapeake Bay). Clinical signs suggested avian botulism, but an ongoing dense Microcystis bloom was present in an impoundment on the island. Enzyme-linked immunosorbent assay (ELISA) analysis of a water sample indicated 6000 ng mL-1 of microcystins (MCs). LC-UV/MS analysis confirmed the presence of MC-LR and a high concentration of an unknown MC congener (m/z 1037.5). The unknown MC was purified and confirmed to be [D-Leu1]MC-LR using NMR spectroscopy, LC-HRMS and LC-MS2, which slowly converted to [D-Leu1,Glu(OMe)6]MC-LR during storage in MeOH. Lyophilized algal material from the bloom was further characterized using LC-HRMS and LC-MS2 in combination with chemical derivatizations, and an additional 24 variants were detected, including MCs conjugated to Cys, GSH and γ-GluCys and their corresponding sulfoxides. Mallard (Anas platyrhynchos) livers were tested to confirm MC exposure. Two broad-specificity MC ELISAs and LC-MS2 were used to measure free MCs, while 'total' MCs were estimated by both MMPB (3-methoxy-2-methyl-4-phenylbutyric acid) and thiol de-conjugation techniques. Free microcystins in the livers (63-112 ng g-1) accounted for 33-41% of total microcystins detected by de-conjugation and MMPB techniques. Free [D-Leu1]MC-LR was quantitated in tissues at 25-67 ng g-1 (LC-MS2). The levels of microcystin varied based on analytical method used, highlighting the need to develop a comprehensive analysis strategy to elucidate the etiology of bird mortality events when microcystin-producing HABs are present.

Examination of contaminant exposure and reproduction of ospreys (Pandion haliaetus) nesting in Delaware Bay and River in 2015.

A study of ospreys (Pandion haliaetus) nesting in the coastal Inland Bays of Delaware, and the Delaware Bay and Delaware River in 2015 examined spatial and temporal trends in contaminant exposure, food web transfer and reproduction. Concentrations of organochlorine pesticides and metabolites, polychlorinated biphenyls (PCBs), coplanar PCB toxic equivalents, polybrominated diphenyl ethers (PBDEs) and other flame retardants in sample eggs were generally greatest in the Delaware River. Concentrations of legacy contaminants in 2015 Delaware Bay eggs were lower than values observed in the 1970s through early 2000s. Several alternative brominated flame retardants were rarely detected, with only TBPH [bis(2-ethylhexyl)-tetrabromophthalate)] present in 5 of 27 samples at <5 ng/g wet weight. No relation was found between p,p'-DDE, total PCBs or total PBDEs in eggs with egg hatching, eggs lost from nests, nestling loss, fledging and nest success. Osprey eggshell thickness recovered to pre-DDT era values, and productivity was adequate to sustain a stable population. Prey fish contaminant concentrations were generally less than those in osprey eggs, with detection frequencies and concentrations greatest in white perch (Morone americana) from Delaware River compared to the Bay. Biomagnification factors from fish to eggs for p,p'-DDE and total PCBs were generally similar to findings from several Chesapeake Bay tributaries. Overall, findings suggest that there have been improvements in Delaware Estuary waterbird habitat compared to the second half of the 20th century. This trend is in part associated with mitigation of some anthropogenic contaminant threats.

Toxicity and hazard of vanadium to mallard ducks (Anas platyrhynchos) and Canada geese (Branta canadensis).

A recent Canada goose (Branta canadensis) die-off at a petroleum refinery fly ash pond in Delaware was attributed to vanadium (V) toxicity. Because of the paucity of V toxicity data for wild birds, a series of studies was undertaken using the forms of V believed to have resulted in this incident. In 7-d single oral dose trials with mallard drakes (Anas platyrhynchos), the estimated median lethal dose (LD50) for vanadium pentoxide was 113 mg/kg body weight, while the LD50 for sodium metavanadate was 75.5 mg/kg. Sodium metavanadate was found to be even more potent (LD50 = 37.2 mg/kg) in male Canada geese. The most distinctive histopathological lesion of both forms of V was lympho-granulocytic enteritis with hemorrhage into the intestinal lumen. Vanadium accumulation in liver and kidney was proportional to the administered dose, and predictive analyses based on these data suggest that V concentrations of 10 microg/g dry weight (dw) in liver and 25 microg/g dw in kidney are associated with mortality (>90% confidence that exposure is >LD50) in mallards acutely exposed to sodium metavanadate. Chronic exposure to increasing dietary concentrations of sodium metavanadate (38.5 to 2651 ppm) over 67 d resulted in V accumulation in liver and kidney (25.2 and 13.6 microg/g dw, respectively), mild intestinal hemorrhage, blood chemistry changes, and evidence of hepatic oxidative stress in mallards, although some of these responses may have been confounded by food avoidance and weight loss. Dietary exposure of mallards to 250 ppm sodium metavanadate for 4 wk resulted in modest accumulation of V in liver and kidney (<5 microg/g dw) and mild intestinal hemorrhage. Based on these data and other observations, it is unlikely that chronic low-level dietary exposure to V poses a direct lethal hazard to wildlife. However, point sources, such as the V-laden fly ash pond encountered by geese at the petroleum refinery in Delaware, may pose a significant hazard to water birds.

Chesapeake Bay fish-osprey (Pandion haliaetus) food chain: Evaluation of contaminant exposure and genetic damage.

From 2011 to 2013, a large-scale ecotoxicological study was conducted in several Chesapeake Bay (USA) tributaries (Susquehanna River and flats, the Back, Baltimore Harbor/Patapsco Rivers, Anacostia/ middle Potomac, Elizabeth and James Rivers) and Poplar Island as a mid-Bay reference site. Osprey (Pandion haliaetus) diet and the transfer of contaminants from fish to osprey eggs were evaluated. The most bioaccumulative compounds (biomagnification factor > 5) included p,p'-dichlorodiphenyldichloroethylene (DDE), total polychlorinated biphenyls (PCBs), total polybrominated diphenyl ethers (PBDEs), and bromodiphenyl ether (BDE) congeners 47, 99, 100, and 154. This analysis suggested that alternative brominated flame retardants and other compounds (methoxytriclosan) are not appreciably biomagnifying. A multivariate analysis of similarity indicated that major differences in patterns among study sites were driven by PCB congeners 105, 128, 156, 170/190, and 189, and PBDE congeners 99 and 209. An integrative redundancy analysis showed that osprey eggs from Baltimore Harbor/Patapsco River and the Elizabeth River had high residues of PCBs and p,p'-DDE, with PBDEs making a substantial contribution to overall halogenated contamination on the Susquehanna and Anacostia/middle Potomac Rivers. The redundancy analysis also suggested a potential relation between PBDE residues in osprey eggs and oxidative DNA damage in nestling blood samples. The results also indicate that there is no longer a discernible relation between halogenated contaminants in osprey eggs and their reproductive success in Chesapeake Bay. Osprey populations are thriving in much of the Chesapeake, with productivity rates exceeding those required to sustain a stable population. Environ Toxicol Chem 2016;35:1560-1575. Published 2016 Wiley Periodicals Inc. on behalf of SETAC. This article is a US Government work and, as such, is in the public domain in the United States of America.

Effects of contaminant exposure on reproductive success of ospreys (Pandion haliaetus) nesting in Delaware River and Bay, USA.

Despite serious water-quality problems and pollutant loading and retention, Delaware River and Bay (USA) provide important wildlife habitat. In 2002, we conducted a comprehensive evaluation of contaminant exposure and reproduction of ospreys (Pandion haliaetus) breeding in Delaware River and Bay. Sample eggs were collected from 39 nests and analyzed for organochlorine pesticides, polychlorinated biphenyls (PCBs), and mercury; a subset of 15 eggs was analyzed for perfluorinated compounds and polybrominated diphenyl ethers (PBDEs). The fate of each nest was monitored weekly. Concentrations of 10 organochlorine pesticides or metabolites, total PCBs, and several toxic PCB congeners were greater (p < 0.05) in eggs collected between the Chesapeake and Delaware Canal (C and D Canal) and Trenton (Delaware River and northern Bay) compared to other sites. Concentrations of p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE; 0.785-3.84 microg/g wet wt) and total PCBs (5.50-14.5 microg/g wet wt) in eggs collected between the C and D Canal and Trenton were similar to levels recently found in the Chesapeake Bay. In all study segments, at least one young fledged from 66 to 75% of nests. Productivity for Delaware Inland Bays (reference area) and southern Delaware Bay was 1.17 and 1.42 fledglings/active nest, respectively; north of the C and D Canal, productivity was 1.00 fledgling/active nest, which is marginally adequate to maintain the population. Using these data, a logistic regression model found that contaminant concentrations (p,p'-DDE, heptachlor epoxide, chlordane and metabolites, and total PCBs) were predictive of hatching success. Several perfluorinated compounds and PBDEs were detected in eggs at concentrations approaching 1 microg/g wet weight. These findings provide evidence that contaminants continue to be a significant stressor on osprey productivity in the northern Delaware River and Bay.

Exposure and food web transfer of pharmaceuticals in ospreys (Pandion haliaetus): Predictive model and empirical data.

The osprey (Pandion haliaetus) is a well-known sentinel of environmental contamination, yet no studies have traced pharmaceuticals through the water-fish-osprey food web. A screening-level exposure assessment was used to evaluate the bioaccumulation potential of 113 pharmaceuticals and metabolites, and an artificial sweetener in this food web. Hypothetical concentrations in water reflecting "wastewater effluent dominated" or "dilution dominated" scenarios were combined with pH-specific bioconcentration factors (BCFs) to predict uptake in fish. Residues in fish and osprey food intake rate were used to calculate the daily intake (DI) of compounds by an adult female osprey. Fourteen pharmaceuticals and a drug metabolite with a BCF greater than 100 and a DI greater than 20 µg/kg were identified as being most likely to exceed the adult human therapeutic dose (HTD). These 15 compounds were also evaluated in a 40 day cumulative dose exposure scenario using first-order kinetics to account for uptake and elimination. Assuming comparable absorption to humans, the half-lives (t1/2 ) for an adult osprey to reach the HTD within 40 days were calculated. For 3 of these pharmaceuticals, the estimated t1/2 in ospreys was less than that for humans, and thus an osprey might theoretically reach or exceed the HTD in 3 to 7 days. To complement the exposure model, 24 compounds were quantified in water, fish plasma, and osprey nestling plasma from 7 potentially impaired locations in Chesapeake Bay. Of the 18 analytes detected in water, 8 were found in fish plasma, but only 1 in osprey plasma (the antihypertensive diltiazem). Compared to diltiazem detection rate and concentrations in water (10/12 detects,

Decadal re-evaluation of contaminant exposure and productivity of ospreys (Pandion haliaetus) nesting in Chesapeake Bay Regions of Concern.

The last large-scale ecotoxicological study of ospreys (Pandion haliaetus) in Chesapeake Bay was conducted in 2000-2001 and focused on U.S. EPA-designated Regions of Concern (ROCs; Baltimore Harbor/Patapsco, Anacostia/middle Potomac, and Elizabeth Rivers). In 2011-2012, ROCs were re-evaluated to determine spatial and temporal trends in productivity and contaminants. Concentrations of p,p'-DDE were low in eggs and below the threshold associated with eggshell thinning. Eggs from the Anacostia/middle Potomac Rivers had lower total PCB concentrations in 2011 than in 2000; however, concentrations remained unchanged in Baltimore Harbor. Polybrominated diphenyl ether flame retardants declined by 40%, and five alternative brominated flame retardants were detected at low levels. Osprey productivity was adequate to sustain local populations, and there was no relation between productivity and halogenated contaminants. Our findings document continued recovery of the osprey population, declining levels of many persistent halogenated compounds, and modest evidence of genetic damage in nestlings from industrialized regions.

Use of the p,p'-DDD: p,p'-DDE concentration ratio to trace contaminant migration from a hazardous waste site.

For approximately 50 years, beginning in the 1920s, hazardous wastes were disposed in an 11-hectare area of the Marine Corps Base (MCB) Quantico, Virginia, USA known as the Old Landfill. Polychlorinated biphenyls (PCBs) and DDT compounds were the primary contaminants of concern. These contaminants migrated into the sediments of a 78-hectare area of the Potomac River, the Quantico Embayment. Fish tissue contamination resulted in the MCB posting signs along the embayment shoreline warning fishermen to avoid consumption. In this paper, we interpret total PCB (t-PCBs) and total DDT (t-DDT, sum of six DDT, DDD, and DDE isomers) data from monitoring studies. We use the ratio of p,p'-DDD to p,p'-DDE concentrations as a tracer to distinguish site-related from regional contamination. The median DDD/DDE ratio in Quantico Embayment sediments (3.5) was significantly higher than the median ratio (0.71) in sediments from nearby Powells Creek, used as a reference area. In general, t-PCBs and t-DDT concentrations were significantly higher in killifish (Fundulus diaphanus) and carp (Cyprinus carpio) from the Quantico Embayment compared with Powells Creek. For both species, Quantico Embayment fish had mean or median DDD/DDE ratios greater than one. Median ratios were significantly higher in Quantico Embayment (4.6) than Powells Creek (0.28) whole body carp. In contrast, t-PCBs and t-DDT in channel catfish (Ictalurus punctatus) fillets were similar in Quantico Embayment and Powells Creek collections, with median ratios of 0.34 and 0.26, respectively. Differences between species may be attributable to movement (carp and killifish being more localized) and feeding patterns (carp ingesting sediment while feeding). We recommend that environmental scientists use this ratio when investigating sites with DDT contamination.

Methodological refinements in the determination of 146 polychlorinated biphenyls, including non-ortho- and mono-ortho-substituted PCBs, and 26 organochlorine pesticides as demonstrated in heron eggs.

A method for the determination of 146 polychlorinated biphenyls (PCBs), including four non-ortho- and eight mono-ortho-substituted congeners, and 26 chlorinated pesticides is described. The method consists of ultrasonic extraction, Florisil cleanup, HPLC fractionation over porous graphitic carbon (PGC), and final determination with GC/ECD, GC/MS, or both. Two PCB congeners (PCB 30 and PCB 161) and two polybromobiphenyls (2,4',5-tribromobiphenyl and 3,3',4,4'-tetrabromobiphenyl) were used as surrogate standards to evaluate the analytical efficiency. Four PCB congeners, PCB 14 and PCB 159 for the first fraction, PCB 61 for the second fraction, and PCB 204 for the third fraction, were used as internal standards to monitor the GC performance. The retention behavior of PCBs and pesticides on the PGC column are discussed. The method was found to be reproducible, effective, and reliable under the operational conditions proposed and was applied successfully to the analysis of individual PCBs and chlorinated pesticides in black-crowned night heron (Nycticorax nycticorax) egg samples.