Understanding diets and structural food webs are keys to the apprehension of ecological communities, upon which conservation and management biology are based. The understanding of grazing and habitat choice for waterfowl is one of the most important topics for avian ecologists today and can, to some degree, be answered by dietary analysis. Droppings collected from four waterfowl, the Eurasian wigeon (Anas penelope), Greylag goose (Anser anser), pink-footed goose (Anser brachyrhynchus) and Barnacle goose (Branta leucopsis) in Vejlerne (Denmark), were analysed microscopically and through eDNA metabarcoding with the use of next generation sequencing (NGS) to accumulate knowledge about the diet of these waterfowl. In total, 120 dropping samples were microscopically analysed, of which the eDNA metabarcoding analysis was done on 79 samples. The prey items were identified according to the taxonomic level of species, and a qualitative method, frequency of occurrence (FO) and FO calculated as a percentage, was used in order to compare the results from the two methods. As neither of the methods was able to encompass all species discovered when combining the two methods, it was concluded in this study that the two methods can support each other in a dietary analysis of waterfowl, but not replace one another.

Philophthalmosis is a zoonotic ocular disease that affects mainly domestic and wild avian species in different parts of the world. However, aspects related to its epizoology in domestic animals are not completely known. In the present study, 47 Muscovy duck (Cairina moschata), 8 mallards (Anas platyrhynchos platyrhynchos) and 6 African geese (Anser cygnoides), free-living in an urban lake in Belo Horizonte, Minas Gerais, Brazil, were evaluated for eye fluke infection. The animals were manually contained, and the conjunctival sac and nictitating membrane were evaluated by direct ophthalmic examination (DOE), in which the fluke burden was estimated by a semiquantitative scale. Moreover, the conjunctival sac lavage (CSL) technique was applied for the search of parasite eggs. When possible, the detected trematodes were carefully removed for fixed for taxonomic identification. In order to characterize the parasite-host-environmental system, malacological collections were performed at the locality, and the collected intermediate host mollusks (Melanoides tuberculata) were evaluated for infection with larval philophthalmids. In addition, wild vertebrates found dead in the locality, including one common gallinule (Gallinula galeata) and two capybaras (Hydrochoerus hydrochaeris), were necropsied for the search of eye flukes. Samples from the developmental stages obtained from the hosts were subjected to morphological and molecular studies (amplification and sequencing of a partial region of the Nad-1 gene). In the DOE, the presence of eye flukes was verified in 67% (4/6), 31% (15/47) and 25% (2/8) in geese, Muscovy ducks and mallards, respectively. In most cases, the birds showed mild parasitism (1-5 trematodes), with no apparent eye damage, characterizing a subclinical infection. In only one goose, which had a high fluke burden (20 trematodes), it was possible to obtain eggs by CSL. Regarding the malacological survey, 73/4545 (1.6%) specimens of M. tuberculata evaluated were found infected with megalurous cercariae, compatible with representatives of the family Philophthalmidae. Additionally, 147 flukes were recovered from a specimen of G. galeata found dead at the lake. The morphological and molecular study of parasites obtained from different hosts evaluated in this study enabled the identification of Philophthalmus gralli, a species native from Asia and introduced in the Americas. Factors such as the wide distribution of the intermediate host and the participation of domestic and wild reservoirs in the parasite transmission chain may favor the occurrence of parasite spillover from wild to domestic birds. Furthermore, the occurrence of the life cycle of P. gralli in an urban waterbody serves as an alert to the possibility of human cases of this zoonotic eye parasite.

Lead poisoning of wild birds by ingestion of lead ammunition occurs worldwide. Histopathological changes in organs of lead-intoxicated birds are widely known, and lead concentration of each organ is measurable using mass spectrometry. However, detailed lead localization at the suborgan level has remained elusive in lead-exposed birds. Here we investigated the detailed lead localization in organs of experimentally lead-exposed ducks and kites by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). In both the ducks and kites, lead accumulated diffusely in the liver, renal cortex, and brain. Lead accumulation was restricted to the red pulp in the spleen. With regard to species differences in lead distribution patterns, it is noteworthy that intensive lead accumulation was observed in the arterial walls only in the kites. In addition, the distribution of copper in the brain was altered in the lead-exposed ducks. Thus, the present study shows suborgan lead distribution in lead-exposed birds and its differences between avian species for the first time. These findings will provide fundamental information to understand the cellular processes of lead poisoning and the mechanisms of species differences in susceptibility to lead exposure.

We utilized lead (Pb) stable isotopes to identify the source of acute Pb poisoning in a Tundra Swan (Cygnus columbianus) and evaluated overall Pb exposure. Upon necropsy, we obtained samples of blood, liver, kidney, heart, thigh, breast, femur, and metallic objects (i.e., fishing sinker, spring and swivels) from the gizzard for Pb isotopic analysis. Pb isotope ratios of blood and soft tissues were essentially identical to the Pb ratios of the sinker, the likely source of acute poisoning. The spring and swivels had lower Pb content and ratios distinct from tissue, suggesting no significant contribution to poisoning. Femur Pb isotopic composition was the most distinct biological sample and indicative of a combination of sources. These results demonstrate isotopic analysis as a viable method for determining the source of acute Pb poisoning, and that Pb isotope ratios in bone most likely record a lifetime-averaged metric of Pb exposure.

The discovery in 1976 of waterfowl as the primary reservoir of influenza A viruses (IAVs) has since spurred decades of waterfowl surveillance efforts by researchers dedicated to understanding the ecology of IAV and its subsequent threat to human and animal health. Here, we employed a multidecade, continental-scale approach of surveillance data to understand trends of seasonal IAV subtype diversity. Between 1976 and 2015, IAVs were detected in 8,427 (10.8%) of 77,969 samples from migratory waterfowl throughout the Central and Mississippi Migratory Flyways in the United States and Canada. A total of 96 hemagglutinin (HA)/neuraminidase (NA) subtype combinations were isolated, which included most HA (H1 to H14) and all 9 NA subtypes. We observed an annual trend of high influenza prevalence, involving a few dominant subtypes, on northern breeding grounds during summer with progressively lowered influenza prevalence, comprised of a highly diverse profile of subtypes, as waterfowl migrate toward southern wintering grounds. Isolates recovered during winter had the highest proportion of mixed and rare HA/NA combinations, indicating increased opportunity for reassortment of IAVs. In addition, 70% of H5 and 49% of H7 IAV isolates were recovered from samples collected during fall and spring, respectively; these are subtypes that can have significant implications for public health and agriculture sectors. Annual cyclical dominance of subtypes on northern breeding grounds is revealed through the longitudinal nature of this study. Our novel findings exhibit the unrealized potential for discovery using existing IAV surveillance data.IMPORTANCE Wild aquatic birds are the primary natural reservoir of influenza A viruses (IAVs) and are therefore responsible for the dispersal and maintenance of IAVs representing a broad range of antigenic and genetic diversity. The aims of IAV surveillance in waterfowl not only relate to understanding the risk of spillover risk to humans, but also to improving our understanding of basic questions related to IAV evolution and ecology. By evaluating several decades of surveillance data from wild aquatic birds sampled along North American migratory flyways, we discovered an annual trend of increasing subtype diversity during southbound migration, peaking on southern wintering grounds. Winter sampling revealed the highest proportion of mixed and rare infections that suggest higher opportunity for spillover. These findings allow improvements to surveillance efforts to robustly capture IAV diversity that will be used for vaccine development and cultivate a more thorough understanding of IAV evolution and persistence mechanisms.