Food supply has been suggested as the main determinant of reproductive success in birds. Riparian species can take advantage of seasonal pulses of both terrestrial and aquatic prey, though aquatic resources are often overlooked in studies of diet and reproductive ecology. This study investigates the flux of both aquatic (mayfly) and terrestrial (caterpillar) prey resources and nestling diet of the Prothonotary Warbler throughout the breeding season in two eastern Virginia sites. One site had significantly higher aquatic prey (mayfly) availability. Nestling diet was generally reflective of prey availability, and nestlings grew faster at the site with high aquatic prey availability. At the site with low aquatic prey availability, nestling growth rates and condition were positively correlated with the amount of aquatic prey in the diet. Our results provide evidence that aquatic subsidies are an important resource for nestlings, and are crucial to understanding the breeding ecology of riparian species.

Populations of many species of migratory birds are declining, and an understanding of how populations are linked between the breeding and nonbreeding grounds is necessary in order to determine drivers of declines. While all current tracking technologies to study the movements of small songbirds are limited by their coarse resolution, tracking birds by measuring stable isotopes in keratin offers an advantage because it requires only a single capture and is cost-effective, which leads to robust sample sizes. While this tracking method is accurate, stable hydrogen isotope values measured in feathers (δ2Hf) are known to be variable within a site, and may be influenced by the hydrology of a site. In this study, we assessed sources of variation in δ2Hf values in a wetland-associated Neotropical migratory bird, the prothonotary warbler (Protonotaria citrea), by comparing δ2Hf values among ages, sexes, years and feather type. We found that age and year significantly influence δ2Hf values, and that differences between primaries and rectrices are statistically significant but generally small. We also tested the accuracy and precision of models to assign prothonotary warblers to their breeding origin using known-origin feathers. We assigned birds in a spatially-explicit manner using an interpolated surface of stable hydrogen values measured in precipitation (δ2Hp), and developed a species-specific calibration equation to account for the offset between δ2Hf and δ2Hp, which explained 51% of the variation in δ2Hf values. We incorporated breeding bird abundance as prior information in our assignment model, and compared two sources of abundance data: the Breeding Bird Survey and a spatio-temporal exploratory model developed with eBird data. The assignment model with no prior information was accurate (83% of birds correctly assigned to their true location of origin), but imprecise (50% of grid cells assigned as likely locations of origin). Incorporating abundance as prior information led to a decrease in accuracy (9-14% of birds correctly assigned) but higher precision (1% of grid cells assigned as likely). We also assigned prothonotary warblers to their breeding origin using feathers collected from across the nonbreeding range. We found that all nonbreeding sampling locations contained a mixture of birds whose origins spanned the majority of the breeding range, with no evidence of strong connectivity between the seasons. In the absence of strong connectivity, the influence of events occurring at one nonbreeding location will be widespread and diffuse across the breeding range. For prothonotary warblers, understanding connectivity can help conservation planners understand how nonbreeding season habitat destruction and other processes influence population dynamics. We recommend that future studies account for age and year variation in δ2Hf values when possible, and continue to examine the trade-off between precision gained and accuracy lost when using abundance as prior information.

As Neotropical migratory birds, Prothonotary Warblers are exposed to parasites in both tropical and temperate regions and may act as dispersal agents between geographic areas. This study identifies the prevalence of Haemosporidia, West Nile Virus (WNV), and St. Louis Encephalitis virus (SLEV) in this species. A total of 71.6% of captured Prothonotary Warblers were infected with Haemosporidia during the 2008 breeding season, and infection prevalence increased throughout the season. This temporal change in prevalence is likely due to infection relapse and transmission of new infections. No correlations between reproductive effort and infection status were observed, nor were any associations between infection prevalence and nest box location identified. WNV and SLEV were present in 37.5% and 6.3% of sampled Prothonotary Warblers, respectively. These results warrant more detailed analyses of pathogen transmission dynamics in this population, physiological mechanisms that affect infection susceptibility, and spatial and temporal trends in infection that may exist.

Algal blooms are directly related to human-caused nutrient enrichment of water bodies. The cyanobacteria Microcystis aeruginosa produces microcystin (MC), a toxin that has been linked with mortalities and illness of many organisms. We show that MC is not constrained by the aquatic-terrestrial ecotone. MC was detected in a primary consumer and emerging aquatic invertebrate (Hexagenia Mayfly), a terrestrial insect and predator of emerging aquatic invertebrates (Tetragnathidae Spider), and a vertebrate consumer (Prothonotary Warbler). Mayfly and spider MC levels varied across the blooming period. MC levels in prothonotary warbler livers varied by age class; nestlings having the highest levels. MC levels decreased in fledglings over time. A more aquatic diet was related to higher MC levels in nestlings at one site and nestling fecal-sacs varied spatially, also indicating that aquatic diet is related to MC consumption. Warbler body condition and growth rate was not related to liver microcystin levels.

Avian malaria is a devastating disease that has decimated numerous bird species. This study sought to identify the vectors of avian malaria at four central Virginia Prothonotary warbler breeding sites. Twenty one thousand mosquitoes were collected and Culex salinarius, Cx. erraticus, and Cx. pipiens/restuans were found to be the dominant species at these sites. Geographic factors, such as crop land and forest type, were determined to be potential indicators for species abundance variation between sites. Of the mosquitoes collected, ninety one (0.4%) were identified as blood fed. The blood fed mosquitoes were found to have fed on avian, mammalian, amphibian, and reptilian hosts and a 12.1% Plasmodium infection rate. Of the non-blood fed mosquito pools tested, Deep Bottom had the highest rate of infection (10.5%). Of the species tested, Cx. salinarius, Cx. erraticus, and Cx. pipiens/restuans were determined to be the most probable vectors of avian malaria the four sites.

Understanding migratory connectivity is essential for assessing the drivers behind population dynamics and for implementing effective management in migratory species. Genetic markers provide a means to describe migratory connectivity, as well as incorporate population genetic analyses, however genetic markers can be uninformative for species with weak genetic structure. In this study, we evaluate range-wide population genetic structure and migratory connectivity in the prothonotary warbler, Protonotaria citrea, a wetland-dependent neotropical migratory songbird, using high-resolution genetic markers. We reveal regional genetic structure between sampling sites in the Mississippi River Valley and the Atlantic Seaboard with overall weak genetic differentiation among populations (FST = 0.0051). By ranking loci by FST and using subsets of the most differentiated genetic markers (200 – 3000), we identify a maximum assignment accuracy (89.7% to site, 94.3% to region) using 600 single nucleotide polymorphisms. We assign samples from unknown origin nonbreeding sites to a breeding region, illustrating weak migratory connectivity between prothonotary warbler breeding and nonbreeding grounds. Our results highlight the importance of using high-resolution markers in studies of migratory connectivity with species exhibiting weak genetic structure. Using similar techniques, studies may begin to describe population genetic structure that was previously undocumented, allowing us to infer the migratory patterns of an increasing number of species.