Novel Coronavirus and Astrovirus in Delaware Bay Shorebirds

Background

Wild birds are an important but to some extent under-studied reservoir for emerging pathogens. We used unbiased sequencing methods for virus discovery in shorebird samples from the Delaware Bay, USA; an important feeding ground for thousands of migratory birds.

Findings

Analysis of shorebird fecal samples indicated the presence of a novel astrovirus and coronavirus. A sanderling sample yielded sequences with distant homology to avian nephritis virus 1, an astrovirus associated with acute nephritis in poultry. A ruddy turnstone sample yielded sequences with homology to deltacoronaviruses.

Conclusions

Our findings highlight shorebirds as a virus reservoir and the need to closely monitor wild bird populations for the emergence of novel virus variants.     

The consequences of climate change at an avian influenza ‘hotspot’

Avian influenza viruses (AIVs) pose significant danger to human health. A key step in managing this threat is understanding the maintenance of AIVs in wild birds, their natural reservoir. Ruddy turnstones (Arenaria interpres) are an atypical bird species in this regard, annually experiencing high AIV prevalence in only one location—Delaware Bay, USA, during their spring migration. While there, they congregate on beaches, attracted by the super-abundance of horseshoe crab eggs. A relationship between ruddy turnstone and horseshoe crab (Limulus polyphemus) population sizes has been established, with a declining horseshoe crab population linked to a corresponding drop in ruddy turnstone population sizes. The effect of this interaction on AIV prevalence in ruddy turnstones has also been addressed. Here, we employ a transmission model to investigate how the interaction between these two species is likely to be altered by climate change. We explore the consequences of this modified interaction on both ruddy turnstone population size and AIV prevalence and show that, if climate change leads to a large enough mismatch in species phenology, AIV prevalence in ruddy turnstones will increase even as their population size decreases.     

Coincident ruddy turnstone migration and horseshoe crab spawning creates an ecological ‘hot spot’ for influenza viruses

Since 1985, avian influenza virus surveillance has been conducted annually from mid-May to early June in charadriiform species from the families Scolopacidae and Laridae (shorebirds and gulls) at Delaware Bay in the northeast United States. The mass migrations of shorebirds, gulls and horseshoe crabs (Limulus polyphemus) coincide at that time, and large numbers of migrating birds pause at Delaware Bay to feed on horseshoe crab eggs deposited at the high-tide line. Influenza viruses are consistently isolated from charadriiform birds at Delaware Bay, at an overall rate approximately 17 times the combined rate of isolation at all other surveillance sites worldwide (490 isolates/9474 samples, 5.2% versus 49 isolates per 15 848 samples, 0.3%, respectively; Proportion test, p < 0.0001). The likelihood of isolating influenza viruses at Delaware Bay is dependent on the presence of ruddy turnstone (Arenaria interpres) at the sampling site (G-test of independence, p < 0.001). The convergence of host factors and environmental factors results in a unique ecological ‘hot spot’ for influenza viruses in Charadriiformes.     

Is the occurrence of avian influenza virus in Charadriiformes species and location dependent?

Birds in the order Charadriiformes were sampled at multiple sites in the eastern half of the continental USA, as well as at Argentina, Chile, and Bermuda, during 1999-2005, and tested for avian influenza virus (AIV). Of more than 9,400 birds sampled, AIV virus was isolated from 290 birds. Although Ruddy Turnstones (Arenaria interpres) comprised just 25% of birds sampled, they accounted for 87% of isolates. Only eight AIV isolations were made from birds at four locations outside of the Delaware Bay, USA, region; six of these were from gulls (Laridae). At Delaware Bay, AIV isolations were predominated by hemagglutinin (HA) subtype H10, but subtype diversity varied each year. These results suggest that AIV infection among shorebirds (Scolopacidae) may be localized, species specific, and highly variable in relation to AIV subtype diversity.     

Molecular Characterization of Subtype H11N9 Avian Influenza Virus Isolated from Shorebirds in Brazil

Migratory aquatic birds play an important role in the maintenance and spread of avian influenza viruses (AIV). Many species of aquatic migratory birds tend to use similar migration routes, also known as flyways, which serve as important circuits for the dissemination of AIV. In recent years there has been extensive surveillance of the virus in aquatic birds in the Northern Hemisphere; however in contrast only a few studies have been attempted to detect AIV in wild birds in South America. There are major flyways connecting South America to Central and North America, whereas avian migration routes between South America and the remaining continents are uncommon. As a result, it has been hypothesized that South American AIV strains would be most closely related to the strains from North America than to those from other regions in the world. We characterized the full genome of three AIV subtype H11N9 isolates obtained from ruddy turnstones (Arenaria interpres) on the Amazon coast of Brazil. For all gene segments, all three strains consistently clustered together within evolutionary lineages of AIV that had been previously described from aquatic birds in North America. In particular, the H11N9 isolates were remarkably closely related to AIV strains from shorebirds sampled at the Delaware Bay region, on the Northeastern coast of the USA, more than 5000 km away from where the isolates were retrieved. Additionally, there was also evidence of genetic similarity to AIV strains from ducks and teals from interior USA and Canada. These findings corroborate that migratory flyways of aquatic birds play an important role in determining the genetic structure of AIV in the Western hemisphere, with a strong epidemiological connectivity between North and South America.     

Complete genome sequence of an H7N3 avian influenza virus isolated from ducks in southern China

We report here the complete genomic sequence of an H7N3 avian influenza virus (AIV) isolate, which was obtained from duck in 1996. This is the first report of this subtype of AIV being isolated from duck in Guangdong of Southern China. Genomic sequence and phylogenetic analyses showed that it was highly homologous with the wild bird virus A/ruddy turnstone/Delaware Bay/135/1996 (H7N3) and that all eight genes of this virus belonged to the North America gene pool. The availability of genome sequences is helpful to further investigations of epidemiology and evolution of AIV between waterfowl and wild birds.     

Avian Influenza Virus (H11N9) in Migratory Shorebirds Wintering in the Amazon Region,Brazil

Aquatic birds are the natural reservoir for avian influenza viruses (AIV). Habitats in Brazil provide stopover and wintering sites for water birds that migrate between North and South America. The current study was conducted to elucidate the possibility of the transport of influenza A viruses by birds that migrate annually between the Northern and Southern Hemispheres. In total, 556 orotracheal/cloacal swab samples were collected for influenza A virus screening using real-time RT-PCR (rRT-PCR). The influenza A virus-positive samples were subjected to viral isolation. Four samples were positive for the influenza A matrix gene by rRT-PCR. From these samples, three viruses were isolated, sequenced and characterized. All positive samples originated from a single bird species, the ruddy turnstone (Arenaria interpres), that was caught in the Amazon region at Caeté Bay, Northeast Pará, at Ilha de Canelas. To our knowledge, this is the first isolation of H11N9 in the ruddy turnstone in South America.     

Avian influenza virus infection dynamics in shorebird hosts

To gain insight into avian influenza virus (AIV) transmission, exposure, and maintenance patterns in shorebirds at Delaware Bay during spring migration, we examined temporal AIV prevalence trends in four Charadriiformes species with the use of serial cross-sectional data from 2000 through 2008 and generalized linear and additive models. Prevalence of AIV in Ruddy Turnstones (Arenaria interpres morinella) increased after arrival, peaked in mid-late May, and decreased prior to departure. Antibody prevalence also increased over this period; together, these results suggested local infection and recovery prior to departure. Red Knots (Calidris canutus rufa), Sanderlings (Calidris alba), and Laughing Gulls (Leucophaeus atricilla) were rarely infected, but dynamic changes in antibody prevalence differed among species. In Red Knots, declining antibody prevalence over the stopover period suggested AIV exposure prior to arrival at Delaware Bay with limited infection at this site. Antibody prevalence was consistently high in Laughing Gulls and low in Sanderlings. Both viral prevalence and antibody prevalence in Sanderlings varied directly with those in turnstones, suggesting virus spillover to Sanderlings. Results indicate that, although hundreds of thousands of birds concentrate at Delaware Bay during spring, dynamics of AIV infection differ among species, perhaps due to differences in susceptibility, potential for contact with AIV at this site, or prior exposure. Additionally, Ruddy Turnstones possibly act as a local AIV amplifying host rather than a reservoir.     

Lines in the mud; revisiting the boundaries of important shorebird areas

Many shorebird populations are declining throughout the world, concurrent with declines and degradation of wetland habitats. Such declines necessitate a more consistent approach towards conserving habitats used by shorebird populations. Individuals of many shorebird species congregate in specific areas during their non-breeding season. Worldwide, non-breeding areas are designated as ‘important’ for shorebird conservation based primarily on the abundance of birds found in an area. However, the boundaries of any area are often defined with incomplete information regarding how shorebirds use that habitat. This paper discusses examples in Australia where improved knowledge of shorebird habitat use led to the identification of very different boundaries of important shorebird areas than those identified originally. We highlight how simple questioning of those who count shorebirds in an area, led to an improved understanding of which areas were apparently used by the same local population of non-breeding shorebirds. Subsequent analysis of available count, recapture and/or home range data of particular shorebird species is needed to verify expert opinion regarding most of these boundaries. We review how enhanced boundaries improve the ability of shorebird monitoring to detect population changes; allow management of shorebird habitats at relevant spatial scales; and lead to appropriate designations of important areas. While the kinds of approaches to boundary setting described here are not new, they are not consistently applied worldwide. We suggest additional guidelines to those produced under the Ramsar Convention in regard to designating important areas. We also call for more studies on the movements of migratory shorebirds during the non-breeding season to direct more consistent boundary setting around important non-breeding habitats used by local populations of migratory shorebirds.     

Detection of avian influenza viruses from shorebirds: evaluation of surveillance and testing approaches

Although influenza A viruses have been isolated from numerous shorebird species (Family: Scolopacidae) worldwide, our understanding of natural history of these viruses in this diverse group is incomplete. Gaining this information can be complicated by sampling difficulties related to live capture, the need for large sample sizes related to a potentially low prevalence of infection, and the need to maintain flexibility in diagnostic approaches related to varied capabilities and resources. To provide information relevant to improving sampling and testing of shorebirds for influenza A viruses, we retrospectively evaluated a combined data set from Delaware Bay, USA, collected from 2000 to 2009. Our results indicate that prevalence trends and subtype diversity can be effectively determined by either direct sampling of birds or indirect sampling of feces; however, the extent of detected subtype diversity is a function of the number of viruses recovered during that year. Even in cases where a large number of viruses are identified, an underestimate of true subtype diversity is likely. Influenza A virus isolation from Ruddy Turnstones can be enhanced by testing both cloacal and tracheal samples, and matrix real-time PCR can be used as an effective screening tool. Serologic testing to target species of interest also has application to shorebird surveillance. Overall, all of the sampling and diagnostic approaches have utility as applied to shorebird surveillance, but all are associated with inherent biases that need to be considered when comparing results from independent studies.     

Identification of significant shorebird areas: thresholds and criteria

Aim Conservation managers designate significant areas for shorebirds based on imperfect data. Significant wetlands for migratory shorebirds have usually been identified on the basis of whether they exceed certain thresholds, defined either by total abundance (usually 20,000 waterbirds) or percentage of a population (usually 1.0%). We evaluate the performance of existing criteria and determine if lowering thresholds would improve shorebird conservation without adding unreasonable numbers of significant sites. Location Australia. Methods We evaluated the best available data, which is used by managers to designate significant areas, to describe the effect of lowering thresholds on the number of significant sites identified and the number of shorebirds these sites support using a range of thresholds in existing criteria. We also investigated factors which may explain interspecific differences evident when lowering thresholds. Results When the threshold for total abundance was lowered from 20,000 to 2000 shorebirds, an additional 45 shorebird areas, holding 65% more shorebirds, were identified. When thresholds for the percentage of a population criterion were lowered from 1.0 to 0.1%, an additional 86 shorebird areas were identified which held 29% more shorebirds. The proportion of a species population counted within wetlands identified as significant by the application of criteria varied widely between species. The percentage of population criterion always identified a network of areas that included more individuals of each species than the total abundance criterion at all threshold levels tested. The percentage of species populations found in networks of significant areas showed greater increase as thresholds were lowered for species that were abundant, widespread and well represented at existing thresholds. Main conclusions Our results suggest lowering thresholds will substantially increase the number of shorebirds in identified significant areas. However, some species will remain under‐represented, partly because of interspecific differences in distribution and inadequate sampling of some shorebird habitats.     

Stable isotope and pen feeding trial studies confirm the value of horseshoe crab Limulus polyphemus eggs to spring migrant shorebirds in Delaware Bay

We used stable isotope (SI) methods in combination with pen feeding trials to determine the importance of eggs of the Atlantic horseshoe crab Limulus polyphemus to migratory fattening of red knots Calidris canutus rufa and ruddy turnstones Arenaria interpres morinella during spring stopover in Delaware Bay. By manifesting measurable fractionation (ca +3‰) and rapid turnover, blood plasma δ¹⁵ nitrogen proved a functional marker for SI diet tracking during the short 3-week stopover. Blood samples from free-ranging knots (3 data sets) and turnstones (1 data set) produced similar convergence of plasma δ¹⁵N signatures with increasing body mass that indicated highly similar diets. Asymptotes deviated slightly (0.3‰ to 0.7‰) from that of captive shorebirds fed a diet of only crab eggs during stopover, thus confirming a strong crab egg-shorebird linkage. The plasma δ¹⁵N crab-egg diet asymptote was enriched ca +4.5‰ and therefore readily discriminated from that of either blue mussels Mytilus edulis or coquina clams Donax variabilis , the most likely alternative prey of knots in Delaware Bay. Crab eggs were highly palatable to captive knots and turnstones which achieved rates of mass gain (3–11 g/d) comparable to that of free-ranging birds. Peak consumption rates during hyperphagic events were 23,940 and 19,360 eggs/bird/d, respectively. The empirical conversions of eggs consumed to body mass gained (5,017 eggs/g for knots and 4,320 eggs/g for turnstones) indicate the large quantities of crab eggs required for the maintenance of these shorebird populations during stopover.     

Seasonal change and habitat selection of shorebird community at the South Yangtze River Mouth and North Hangzhou Bay, China

Coastal regions are important habitats for migratory shorebirds. The aim of the study is to understand habitat use by migratory shorebirds and to develop a conservation strategy in the sustainable use of wetlands. From March 2004 to January 2005, we conducted a seasonal shorebirds census in ten coastal habitats along the South Yangtze River mouth and North Hangzhou Bay, simultaneously examining the relative seasonal abundance of shorebirds and their spatial distribution. A total of 25 species were identified, the dominant seasonal species were Great Knot (Calidris tenuirostris), Sharp-tailed Sandpiper (Calidris alpine) and Red-necked Stint (Calidris ruficollis) in spring; Kentish Plover (Charadrius alexandrinus), Common Greenshank(Tringa nebularia) and Lesser Sand Plover (Charadrius mongolus) in summer; Kentish Plover, Red-necked Stint and Common Greenshank in autumn; Dunlin(Calidris alpine), Kentish Plover and Marsh Sandpiper (Tringa stagnatilis) in winter. These species accounted for more than 85% of the total shorebirds. The numbers of shorebirds counted was highest in spring and then in autumn, winter and summer respectively. Among the four seasons, there were few significant differences in the number of bird species between the sites outside the seawall (intertidal mudflat) and the sites inside the seawall (artificial wetland), but the average density of shorebirds was obviously different. The habitat-selection analysis of the environmental factors (outside and inside the seawall) impacting on the shorebird community was made in the 10 study sites with Canonical Correspondence Analysis. The study results indicated that: (1) Outside the seawall, the widths of the total intertidal mudflat and bare mudflat were the key factors affecting the shorebirds; the proportion of bulrush (Scirpusmariquete) covering and supertidal mudflat width had a positive correlation with the abundance of birds, while human disturbance and the proportion of both reed (Phragmites communis) and smooth cord-grass (Spartina alterniflora) covering in total surveyed areas had negative impacts on bird numbers; (2) Inside the seawall, the proportions of areas with shallow water and mudflats occupying the total surveyed area were key factors influencing the number of birds; the size of the bulrush area should have a positive impact on the appearance of shorebirds. Habitats with heavy human disturbance, dense reed and smooth cord-grass or a high water level were not conducive to be inhabited by shorebirds.     

Intestinal Microbiota and Species Diversity of Campylobacter and Helicobacter spp. in Migrating Shorebirds in Delaware Bay

Using 16S rRNA gene sequencing analysis, we examined the bacterial diversity and the presence of opportunistic bacterial pathogens (i.e., Campylobacter and Helicobacter) in red knot (Calidris canutus; n = 40), ruddy turnstone (Arenaria interpres; n = 35), and semipalmated sandpiper (Calidris pusilla; n = 22) fecal samples collected during a migratory stopover in Delaware Bay. Additionally, we studied the occurrence of Campylobacter spp., enterococci, and waterfowl fecal source markers using quantitative PCR (qPCR) assays. Of 3,889 16S rRNA clone sequences analyzed, the bacterial community was mostly composed of Bacilli (63.5%), Fusobacteria (12.7%), Epsilonproteobacteria (6.5%), and Clostridia (5.8%). When epsilonproteobacterium-specific 23S rRNA gene clone libraries (i.e., 1,414 sequences) were analyzed, the sequences were identified as Campylobacter (82.3%) or Helicobacter (17.7%) spp. Specifically, 38.4%, 10.1%, and 26.0% of clone sequences were identified as C. lari (>99% sequence identity) in ruddy turnstone, red knot, and semipalmated sandpiper clone libraries, respectively. Other pathogenic species of Campylobacter, such as C. jejuni and C. coli, were not detected in excreta of any of the three bird species. Most Helicobacter-like sequences identified were closely related to H. pametensis (>99% sequence identity) and H. anseris (92% sequence identity). qPCR results showed that the occurrence and abundance of Campylobacter spp. was relatively high compared to those of fecal indicator bacteria, such as Enterococcus spp., E. faecalis, and Catellicoccus marimammalium. Overall, the results provide insights into the complexity of the shorebird gut microbial community and suggest that these migratory birds are important reservoirs of pathogenic Campylobacter species.     

Seasonal change and habitat selection of shorebird community at the South Yangtze River Mouth and North Hangzhou Bay, China

Coastal regions are important habitats for migratory shorebirds. The aim of the study is to understand habitat use by migratory shorebirds and to develop a conservation strategy in the sustainable use of wetlands. From March 2004 to January 2005, we conducted a seasonal shorebirds census in ten coastal habitats along the South Yangtze River mouth and North Hangzhou Bay, simultaneously examining the relative seasonal abundance of shorebirds and their spatial distribution. A total of 25 species were identified, the dominant seasonal species were Great Knot (Calidris tenuirostris), Sharp-tailed Sandpiper (Calidris alpine) and Red-necked Stint (Calidris ruficollis) in spring; Kentish Plover (Charadrius alexandrinus), Common Greenshank(Tringa nebularia) and Lesser Sand Plover (Charadrius mongolus) in summer; Kentish Plover, Red-necked Stint and Common Greenshank in autumn; Dunlin(Calidris alpine), Kentish Plover and Marsh Sandpiper (Tringa stagnatilis) in winter. These species accounted for more than 85% of the total shorebirds. The numbers of shorebirds counted was highest in spring and then in autumn, winter and summer respectively. Among the four seasons, there were few significant differences in the number of bird species between the sites outside the seawall (intertidal mudflat) and the sites inside the seawall (artificial wetland), but the average density of shorebirds was obviously different. The habitat-selection analysis of the environmental factors (outside and inside the seawall) impacting on the shorebird community was made in the 10 study sites with Canonical Correspondence Analysis. The study results indicated that: (1) Outside the seawall, the widths of the total intertidal mudflat and bare mudflat were the key factors affecting the shorebirds; the proportion of bulrush (Scirpus×mariquete) covering and supertidal mudflat width had a positive correlation with the abundance of birds, while human disturbance and the proportion of both reed (Phragmites communis) and smooth cord-grass (Spartina alterniflora) covering in total surveyed areas had negative impacts on bird numbers; (2) Inside the seawall, the proportions of areas with shallow water and mudflats occupying the total surveyed area were key factors influencing the number of birds; the size of the bulrush area should have a positive impact on the appearance of shorebirds. Habitats with heavy human disturbance, dense reed and smooth cord-grass or a high water level were not conducive to be inhabited by shorebirds.     

Wind conditions affect stopover decisions and fuel stores of shorebirds migrating through the south Yellow Sea

We assessed the effects of wind conditions on stopover decisions and fuel stores of migratory shorebirds at Chongming Dongtan in the south Yellow Sea along the East Asian–Australasian Flyway. In spring and autumn, wind directions differed among altitudes and wind speed generally increased with altitude. Numbers of shorebirds were related to wind effects at low altitudes (on the ground and at 300 and 800 m above the ground), wind effects at 300 m being the best predictor of shorebird numbers. In spring, total number of shorebirds and numbers of the four most abundant shorebird species were negatively related to wind assistance at low altitudes, more birds departing when tailwinds prevailed and more arriving when headwinds prevailed. In autumn, however, total number of shorebirds and numbers of the four most abundant species were positively related to wind assistance at low altitudes, more birds departing and more arriving with tailwinds than with headwinds. When tailwinds prevailed, the number of arriving birds was higher than the number of departing birds. The fuel stores of captured shorebirds, represented by their body mass, was related to wind effects and change in wind conditions between two consecutive days in both spring and autumn, captured birds being heavier when headwinds prevailed than in tailwind conditions, and when the wind conditions became less favourable for flight between two consecutive days. Our results suggest that wind conditions affect stopover decisions and fuel stores, and thus the optimal migration and fuel deposition strategies of migratory shorebirds.     

Influenza A Virus Migration and Persistence in North American Wild Birds

Wild birds have been implicated in the emergence of human and livestock influenza. The successful prediction of viral spread and disease emergence, as well as formulation of preparedness plans have been hampered by a critical lack of knowledge of viral movements between different host populations. The patterns of viral spread and subsequent risk posed by wild bird viruses therefore remain unpredictable. Here we analyze genomic data, including 287 newly sequenced avian influenza A virus (AIV) samples isolated over a 34-year period of continuous systematic surveillance of North American migratory birds. We use a Bayesian statistical framework to test hypotheses of viral migration, population structure and patterns of genetic reassortment. Our results reveal that despite the high prevalence of Charadriiformes infected in Delaware Bay this host population does not appear to significantly contribute to the North American AIV diversity sampled in Anseriformes. In contrast, influenza viruses sampled from Anseriformes in Alberta are representative of the AIV diversity circulating in North American Anseriformes. While AIV may be restricted to specific migratory flyways over short time frames, our large-scale analysis showed that the long-term persistence of AIV was independent of bird flyways with migration between populations throughout North America. Analysis of long-term surveillance data provides vital insights to develop appropriately informed predictive models critical for pandemic preparedness and livestock protection.     

The costs of migration: Injuries in migratory waterbirds along the west coast of India

The long distant, transcontinental migration of shorebirds entails many well identified costs in terms of time, energy, and direct mortality risk. Injuries from debris or from human structures and activities were observed as the major reasons for the direct mortality of shorebirds during migration worldwide. We recorded injured birds in major coastal wetlands of Kerala, for a period of 15 years from 2005 to 2019. The injured birds were observed in 9 different sites in various districts of Kerala. The highest instances of injuries were observed in Kadalundi-Vallikunnu Community Reserve, the major wintering and stop over site of migrant shorebirds in the west coast of India. During the study period, fifty-eight individuals of shorebirds belonging to four families were found to be injured. The highest proportion of injuries was recorded among the families Scolopacidae and Charadriidae comprising long distance migrant shorebird species and the lowest among Laridae and Ardeidae. We recommend that environmental authorities pay special attention to minimize anthropogenic debris along the flyways used by migratory birds thereby reducing the risk of injuries to some of these species. Proactive measures such as removal of discarded fishing gear or plastic debris from wintering areas as well as stopover areas could greatly reduce injuries in migratory birds arising from anthropogenic sources.     

Complete mitochondrial DNA genome sequences show that modern birds are not descended from transitional shorebirds

To test the hypothesis put forward by Feduccia of the origin of modern birds from transitional birds, we sequenced the first two complete mitochondrial genomes of shorebirds (ruddy turnstone and blackish oystercatcher) and compared their sequences with those of already published avian genomes. When corrected for rate heterogeneity across sites and non-homogeneous nucleotide compositions among lineages in maximum likelihood (ML), the optimal tree places palaeognath birds as sister to the neognaths including shorebirds. This optimal topology is a re-rooting of recently published ordinal-level avian trees derived from mitochondrial sequences. Using a penalized likelihood (PL) rate-smoothing process in conjunction with dates estimated from fossils, we show that the basal splits in the bird tree are much older than the Cretaceous-Tertiary (K-T) boundary, reinforcing previous molecular studies that rejected the derivation of modern birds from transitional shorebirds. Our mean estimate for the origin of modern birds at about 123 million years ago (Myr ago) is quite close to recent estimates using both nuclear and mitochondrial genes, and supports theories of continental break-up as a driving force in avian diversification. Not only did many modern orders of birds originate well before the K-T boundary, but the radiation of major clades occurred over an extended period of at least 40 Myr ago, thus also falsifying Feduccia's rapid radiation scenario following a K-T bottleneck.     

Niche dynamics of shorebirds in Delaware Bay: Foraging behavior,habitat choice and migration timing

Niche differentiation through resource partitioning is seen as one of the most important mechanisms of diversity maintenance contributing to stable coexistence of different species within communities. In this study, I examined whether four species of migrating shorebirds, dunlins (Calidris alpina), semipalmated sandpipers (Calidris pusilla), least sandpipers (Calidris minutilla) and short-billed dowitchers (Limnodromus griseus), segregate by time of passage, habitat use and foraging behavior at their major stopover in Delaware Bay during spring migration. I tested the prediction that most of the separation between morphologically similar species will be achieved by differential migration timing. Despite the high level of overlap along observed niche dimensions, this study demonstrates a certain level of ecological separation between migrating shorebirds. The results of analyses suggest that differential timing of spring migration might be the most important dimension along which shorebird species segregate while at stopover in Delaware Bay. Besides differences in time of passage, species exhibited differences in habitat use, particularly least sandpipers that foraged in vegetated areas of tidal marshes more frequently than other species, as well as short-billed dowitchers that foraged in deeper water more often than small sandpipers did. Partitioning along foraging techniques was less prominent than segregation along temporal or microhabitat dimensions. Such ranking of niche dimensions emphasizes significance of temporal segregation of migratory species – separation of species by time of passage may reduce the opportunity for interspecific aggressive encounters, which in turn can have positive effects on birds' time and energy budget during stopover period.