Using naturally shed feathers for individual identification, genetic parentage analyses, and population monitoring in an endangered Eastern imperial eagle (Aquila heliaca) population from Kazakhstan

Genetic analyses on noninvasively collected samples have revolutionized how populations are monitored. Most noninvasive monitoring studies have used hair or scat for individual identification of elusive mammals, but here we utilize naturally shed feathers. The Eastern imperial eagle (EIE) is a species of conservation concern throughout Central Asia and, like most raptors, EIEs are inherently challenging to study because adults are difficult to capture and band using conventional techniques. Over 6 years, we noninvasively collected hundreds of adult feathers and directly sampled EIE chicks at a national nature reserve in Kazakhstan. All samples were genetically sexed and genotyped at a suite of microsatellite loci. Genetically profiled adult feathers identified and monitored the presence of individual eagles over time, enabling us to address a variety of issues related to the biology, demography, and conservation of EIEs. Specifically, we characterized (i) the genetic mating system, (ii) relatedness among mated pairs, (iii) chick sex ratios, and (iv) annual turnover in an adult breeding population. We show that EIEs are genetically monogamous and furthermore, there is no apparent relatedness-based system of mate choice (e.g. inbreeding avoidance). Results indicate that annual adult EIE survivorship (84%) is lower than expected for a long-lived raptor, but initial analyses suggest the current reproductive rate at our study site is sufficient to maintain a stable breeding population. The pristine habitat at our study site supports an EIE population that is probably the most demographically robust in the world; thus, our results caution that populations in marginal habitats may not be self-sustaining.     

The feeding ecology of Bonelli’s eagle (<Emphasis Type="Italic">Aquila fasciata</Emphasis>) floaters in southern Spain: implications for conservation

Although many studies have investigated the feeding habits of Palaearctic raptors, few have analysed non-breeding populations during dispersal. Bonelli’s eagle (Aquila fasciata), a threatened species in Western Europe, has a relatively long and critical dispersal period. We studied feeding habits, prey selection, and the influence of prey density on floater abundance in this species during its dispersal period in southern Spain. Differences were found between the diet of floaters and that of the closest breeding populations. Diet diversity was rather low for floaters, with European wild rabbits (Oryctolagus cuniculus) being the main prey and the only prey positively selected. Moreover, the number of floater Bonelli’s eagles observed in the dispersal areas was positively associated with rabbit abundance. Other prey included red-legged partridges (Alectoris rufa) and pigeons (Columba sp.). We propose measures to match suitable prey availability for floater eagles in settlement areas and the use of such areas as human hunting fields.     

Tetranucleotide microsatellites for aquila and haliaeetus eagles

A unique community of four syntopic eagle species exists in north‐central Kazakhstan. Questions about behaviour and genetics in these four species would benefit from the development of microsatellite markers. We isolated eight polymorphic microsatellite repeats (AAAG)_n from the eastern imperial eagle (Aquila heliaca) genome using a hybridization enrichment technique. These loci revealed moderate diversity in a local population of eastern imperial eagles (observed heterozygosity 0.26–0.78), and were also polymorphic in steppe eagles (A. nipalensis) and white‐tailed sea eagles (Haliaeetus albicilla). These primers may be polymorphic in other species of Aquila and Haliaeetus eagles.     

Positive assortative pairing by plumage colour in Spanish imperial eagles

Intraspecific variation in plumage may have a functional significance in mate choice. As a result, breeding birds are often paired assortatively with respect to colour. However, whether this within-pair correlation is a result of homotypic preference whereby individuals pick a mate that looks like themselves, or directional preference whereby all individuals have the same phenotypic preference, is often unknown. Using data collected between 1989 and 2006, we describe intraspecific variation in the striking white feathers on the leading edge of the wing, lesser coverts and mantle of 144 Spanish imperial eagles (Aquila adalberti), one of the most endangered birds of prey in the world. Females had, on average, more white than males, and pairs mated in a positive assortative fashion. Coloration was not related to age of the bird, food supply (i.e., territory quality) or breeding productivity. Our results are most consistent with the process of homotypic-preference assortative mating, and this may be a result of sexual imprinting, and function as a mechanism to optimize the degree of outbreeding. This pattern of mate selection may explain the rapid evolutionary divergence of A. adalberti from the continental population of imperial eagle A. heliaca.     

Species identification of birds through genetic analysis of naturally shed feathers

Genetic analysis of noninvasively collected bird feathers is of growing importance to avian ecology; however, most genetic studies that utilize feathers make no mention of the need to verify their species of origin. While plumage patterns and collection location often are indicative of species identity, broad‐scale feather collections may require definitive species identification prior to analysis. Genetic species identification has been applied to noninvasively collected samples from a wide range of taxa but, to date, these techniques have not been widely used on bird feathers. Here, we develop and test a polymerase chain reaction (PCR)‐based technique for identifying eastern imperial eagle (Aquila heliaca) samples among a vast number of noninvasively collected feathers. Species identification is accomplished by amplifying a fragment of the mitochondrial cytochrome c oxidase I gene, then digesting that fragment with a restriction enzyme. The resulting species‐specific restriction fragment length polymorphisms (RFLPs) are easily visualized by gel electrophoresis. We tested this PCR‐RFLP assay on over 300 individuals that had been genetically identified from noninvasively collected feathers and demonstrated that the assay is both reliable and robust for DNA of low quality and quantity. The genetic methods of species identification used to develop this assay can readily be applied to other bird assemblages, making them particularly relevant to a broad range of future avian research.     

Natural expansion versus translocation in a previously human‐persecuted bird of prey

Many threatened species in Europe have been expanding their distributions during recent decades owing to protection measures that overcome historical human activity that has limited their distributions. Range expansion has come about via two processes, natural expansion from existing range and reintroductions to new ranges. Reintroductions may prove to be a better way to establish populations because individuals are less subject to competitive relationships lowering breeding success than individuals expanding from existing populations. Whether this is true, however, remains uncertain. We compared success of breeding pairs of an expanding and a reintroduced population of spanish imperial eagles monitored for over 15 years in the south of Spain. We found significant differences in productivity between breeding pairs of each population. Newly established territories in reintroduction areas were almost three times more productive than new territories established as individuals expanded out from an existing population. We conclude that among these eagle populations reintroduced to new areas may fare as well or better than individuals expanding out form existing populations.     

Availability of optimal‐sized prey affects global distribution patterns of the golden eagle Aquila chrysaetos

Climate and landscape change are expected to significantly affect trophic interactions, which will especially harm top predators such as the golden eagle Aquila chrysaetos. Availability of optimal prey is recognized to influence reproductive success of raptors on a regional scale. For the golden eagle, medium‐sized prey species between 0.5 and 5 kg are widely considered to be optimal prey during the breeding season, whereas smaller and larger species are deemed as energetically sub‐optimal. However, knowledge about the effects of optimal prey availability is still scarce on larger scales. To decrease this apparent knowledge gap, we combined biogeographical information on range margins with information about the foraging behaviour and reproductive success of golden eagles from 67 studies spanning the Northern Hemisphere. We hypothesized that availability of optimal prey will affect foraging behaviour and breeding success and, thus, distribution patterns of the golden eagle not only on a local but also on a continental scale. We correlated the diet breadth quantifying foraging generalism, breeding success and proportions of small (< 0.5 kg), medium (0.5–5 kg) and large‐sized (> 5 kg) prey species within the diet with the minimum distance of the examined eagles to the actual species distribution boundary. Closer to the range edge, we observed decreased proportions of medium‐sized prey species and decreasing breeding success of golden eagles. Diet breadth as well as proportions of small and large‐sized prey species increased, however, towards the range edge. Thus, availability of optimal‐sized prey species seems to be a crucial driver of foraging behaviour, breeding success and distribution of golden eagles on a continental scale. However, underlying effects of landscape characteristics and human influence on optimal prey availability has to be investigated in further large‐scale studies to fully understand the major threats facing the golden eagle and possibly other large terrestrial birds of prey.     

Survey for hemoparasites in imperial eagles (Aquila heliaca), steppe eagles (Aquila nipalensis), and white-tailed sea eagles (Haliaeetus albicilla) from Kazakhstan

Prevalence of hemoparasites has been investigated in many avian species throughout Europe and North America. Basic hematologic surveys are the first step toward evaluating whether host-parasite prevalences observed in North America and Europe occur elsewhere in the world. We collected blood smears from 94 nestling imperial eagles (Aquila heliaca), five nestling steppe eagles (Aquila nipalensis), and 14 nestling white-tailed sea eagles (Haliaeetus albicilla) at Naurzum Zapovednik (Naurzum National Nature Reserve) in Kazakhstan during the summers of 1999 and 2000. In 1999, six of 29 imperial eagles were infected with Lencocytozoon toddi. Five of 65 imperial eagles and one of 14 white-tailed sea eagle were infected with L. toddi in 2000. Furthermore, in 2000, one of 65 imperial eagles was infected with Haemoproteus sp. We found no parasites in steppe eagles in either year, and no bird had multiple-species infections. These data are important because few hematologic studies of these eagle species have been conducted.     

Behavioral evidence of hunting and foraging techniques by a top predator suggests the importance of scavenging for preadults

Scavenging may be a regular feeding behavior for some facultative raptor species occupying low quality habitats and/or with little experience in hunting techniques. However, its importance has been largely underestimated due to methodological limitations in identifying the real proportion in the diet. Here, through direct observations, we assessed the hunting and foraging success of the threatened Spanish imperial eagle Aquila adalberti determining the influence of age, sex, breeding status, habitat quality, prey type, and landscape characteristics. From 465 observations, Spanish imperial eagles used hunting in flight (42%), scavenging (30%), hunting from a perch (16%) and kleptoparasitism (12%). Our model suggests that Prey size and Prey type best explain hunting success, followed by Landscape and Sex. Our findings suggest that Spanish imperial eagles increase hunting success with age, with scavenging and kleptoparasitism regularly used as juveniles. The absence of relationships with any of the variables considered suggests that kleptoparasitism is an opportunistic behavior used sporadically. Scavenging is also independent of habitat quality and landscape characteristics. Accordingly, low prey density is not a driver of carrion use for preadult individuals, suggesting that a lack of hunting ability obliges this age‐class to use this alternative feeding technique regularly. As a result, the threatened Spanish imperial eagle population is also prone to mortality related to the illegal use of poison baits and, potentially, veterinary drugs (i.e., diclofenac).     

Sexual aggression by intruders in hooded crow <Emphasis Type="Italic">Corvus cornix</Emphasis>

The hooded crow Corvus cornix is a west Palaearctic, solitary nesting, monogamous corvid. In the breeding season, populations are characterized by a social organization wherein breeding pairs are territorial and non-breeding individuals, called floaters, live in flocks. During a study of the breeding ecology of the hooded crow, conducted in a protected flooded area, we monitored nests with video cameras. We recorded two separate incidents when intruders attacked a female at the nest. We believe that she remained in the nest in order to prevent the strangers cannibalizing the nestlings by mantling over the brood. The spatio-temporal occurrence of these attacks suggests that the observed behaviour is intraspecific sexual aggression wherein non-breeding males mounted an immobilized female.     

Genetic structure and phylogeography of a European flagship species,the white‐tailed sea eagle Haliaeetus albicilla

We analysed 120 white‐tailed sea eagles Haliaeetus albicilla from eastern (Poland and Estonia) and southeastern (Serbian Danube population) Europe for genetic variability and structuring at the mitochondrial control region and seven nuclear microsatellite loci. We combined this new dataset with sequence and genotype data from previous analyses covering Greenland and Eurasia (total sample sizes of 420 and 186 individuals for mtDNA and microsatellites, respectively) to address the following questions: 1) does the large eastern population in Europe add significantly to the species‘ overall genetic diversity? 2) Do the new sequence data match the clinal distribution pattern (west to east) of the two major mtDNA lineages? 3) Does the preliminary hypothesis of two nuclear genetic clusters recently found in this species hold for the whole of Europe, and do these clusters show a geographic pattern? Our results confirmed Europe as a stronghold of genetic diversity in white‐tailed sea eagles, and the east of the continent contributed disproportionately to this, the reason being the admixture of eagles with different genetic background. As hypothesised, both mitochondrial lineages were recovered also in eastern Europe, but the globally more eastern lineage was dominant. The presence of two microsatellite clusters was also confirmed, and these groups, too, show a non‐random geographic distribution, with, except for Poland, a high proportion of ‘eastern‐type’ eagles in the populations of east–central and eastern Europe.     

Causes of death in different areas for Bonelli's Eagle Hieraaetus fasciatus in Spain

The Spanish Bonelli's Eagle populations have decreased markedly because of high mortality. We recorded 424 cases of dead eagles between 1990 and 1998 in Spain which after cross-comparison corresponded to 377 individuals. Electrocution (55% of deaths), followed by direct persecution (26%) were the main causes of death. No differences in the cause of death were found between sexes. Non-adult eagles mostly died of electrocution whereas adults were mainly the victims of persecution. A log-linear model showed that these differences were associated with a difference in the spatial distribution of age classes, rather than to age or experience per se. Persecution was the main cause of death in breeding areas and electrocution in non-breeding areas. There were differences between regions: electrocution was the main cause in Catalonia and Central Spain (50% and 86% respectively) whereas direct persecution was the main cause in Levant and Northern Spain (52% and 43% respectively). We recommend that steps are taken in order to reduce eagle mortality, taking into account the differences between regions and areas.     

Genetic variability in the Iberian imperial eagle (Aquila adalberti) demonstrated by RAPD analysis

RAPD analysis was used to estimate the genetic diversity in an Iberian imperial eagle (Aquila adalberti) population, one of the most threatened bird species in the world. Forty-five of 60 arbitrarily designed primers amplified 614 loci in 25 individual eagles, 59.7% of which were polymorphic. In contrast to the traditional allozyme analysis performed in a previous study, the RAPD method has revealed a high level of heterozygosity in this species (H = 0.267+/-0.008). The genetic distances estimated between 25 eagles can serve to establish more adequate mating in order to preserve genetic variability. Conservation efforts being carried out in Spain in this species might be successful based on the results obtained in the present work.     

Genetic divergence between colonies of Flesh-footed Shearwater <Emphasis Type="Italic">Ardenna carneipes</Emphasis> exhibiting different foraging strategies

Increasing evidence suggests foraging segregation as a key mechanism promoting genetic divergence within seabird species. However, testing for a relationship between population genetic structure and foraging movements among seabird colonies can be challenging. Telemetry studies suggest that Flesh-footed Shearwater Ardenna carneipes that breed at Lord Howe Island or New Zealand, versus southwestern Australia or Saint-Paul Island in the Indian Ocean, migrate to different regions (North Pacific Ocean and northern Indian Ocean, respectively) during the non-breeding season, which may inhibit gene flow among colonies. In this study, we sequenced a 858-base pair mitochondrial region and seven nuclear DNA fragments (352–654 bp) for 148 individuals to test genetic differentiation among colonies of Flesh-footed Shearwaters. Strong genetic divergence was detected between Pacific colonies relative to those further West. Molecular analysis of fisheries’ bycatch individuals sampled in the Sea of Japan indicated that individuals from both western and eastern colonies were migrating through this area, and hence the apparent segregation of the non-breeding distribution based on telemetry is invalid and cannot contribute to the population genetic structure among colonies. The genetic divergence among colonies is better explained by philopatry and evidence of differences in foraging strategies during the breeding season, as supported by the observed genetic divergence between Lord Howe Island and New Zealand colonies. We suggest molecular analysis of fisheries’ bycatch individuals as a rigorous method to identify foraging segregation, and we recommend the eastern and western A. carneipes colonies be regarded as different Management Units.     

The ‘chaotic’ flight feather moult of the Steppe Buzzard Buteo buteo vulpinus

Steppe Buzzards breed in Eurasia and spend the non-breeding season in Africa. Adults moult some flight feathers during the breeding season and some during the non-breeding season. Moult is arrested during migration. The extent of moult of flight feathers in adults is highly variable between individuals in southern Africa, with the renewal of two primaries, three secondaries and five rectrices as the most frequently encountered pattern. Time spent on the non-breeding grounds in South Africa is too short to allow for a sequential moult. Moult of flight feathers is restricted to the almost synchronous dropping of a number of feathers upon arrival, with few being replaced subsequently. Any of the flight feathers can be replaced in southern Africa, and the pattern of renewal in primaries and secondaries cannot be distinguished from random. Tail feathers are replaced in an alternating (transilient) pattern. Moult in the non-breeding areas may primarily be complementary to moult on the breeding grounds, but these two partial moults per year are insufficient to renew all flight feathers annually. Middle secondaries and central tail feathers are regularly carried over to a third moult, but this is rare for primaries.     

Floater interference reflects territory quality in the Spanish Imperial Eagle Aquila adalberti: a test of a density‐dependent mechanism

We report on an 11‐year study of floater interference in a population of Spanish Imperial Eagles Aquila adalberti. We analysed changes over the years in the productivity of 15 territories to test predictions of two hypotheses of density‐dependent productivity in relation to the presence of floaters (birds without territories). According to the ‘interference' hypothesis, the frequency of intrusion by floaters increases with density, resulting in a decrease in productivity. Thus, in a high‐density population a negative relationship between floater intrusions and productivity of the territory is expected. In contrast, under the ‘habitat heterogeneity' hypothesis, as density increases a higher proportion of individuals is forced to occupy lower quality habitats. Support of this hypothesis requires that floaters detect differences in quality among territories and preferentially visit the better quality territories. Consequently, a positive relationship between floater intrusions and productivity is expected. Results showed that floaters tended to visit their natal area at the beginning of the breeding season. Among floater Eagles, males made significantly more intrusions per day than did females, but females stayed in the natal population for longer each year than males. Floater intrusions and productivity were highly positively correlated, supporting the ‘habitat heterogeneity' hypothesis; individuals were apparently able to assess the quality of a territory and, at the frequencies observed, their interference with the breeding pair had no obvious negative effect on productivity.     

Limited dispersal by Nazca boobies Sula granti

We documented natal and breeding dispersal at several spatial scales by Galápagos Nazca boobies Sula granti , a wide-ranging pelagic seabird. We found exceptionally low degrees of both types of dispersal despite these birds' vagility. Median natal dispersal distances were 26 m and 105 m for males and females, respectively. Median breeding dispersal distances for both sexes were 0 m. No natal or breeding dispersals occurred from our study site at Punta Cevallos, Isla Española to six other colonies in the Galápagos, but we did document four long-distance natal dispersals from Punta Cevallos to islands near the South American coast. Recaptures and dead recoveries of ringed birds showed long distance non-breeding movements to the Central American coast and elsewhere in the eastern Pacific, contrasting with the very limited dispersal to breeding sites.     

The effects of habitat quality and female size on the productivity of the lesser spotted eagle Aquila pomarina in the light of the alternative prey hypothesis

Habitat quality is an important but insufficiently understood concept in ecology and conservation biology, due to geographic and temporal variation as well as interaction with individual quality. In 1994–2002, we studied the Estonian population of the lesser spotted eagle Aquila pomarina in order to (1) explore the relative contributions of habitat and female size in reproductive success; (2) check for a switch to alternative prey in vole‐poor years and the relevant variation in annual habitat quality as confirmed in the common buzzard Buteo buteo in the same area. We measured five landscape variables, the number of neighbouring conspecifics and the relative size of the female according to large moulted feathers in 77 nesting territories, and related this to the eagles’ productivity in vole‐rich and vole‐poor years. Nesting lesser spotted eagles benefited from heterogeneous landscapes and suffered from the neighbourhood of conspecifics. There was no evidence that different‐sized females used different habitats. In general, female size was positively related to productivity in vole‐poor but not vole‐rich years, but in the presence of competitors, large size appeared to be disadvantageous. The mean annual productivity of the eagle was well correlated with that of the buzzard, both having peaks after every three years. In contrast to the buzzard, the share of voles in the eagle's diet and its habitat quality did not differ significantly between good and poor years. We concluded that despite a superficial ecological similarity to the buzzard, the lesser spotted eagle did not behave as predicted by the alternative prey hypothesis, but the study confirmed that annual variation in prey utilization and relative habitat quality are parts of the same functional response. Non‐switching to alternative prey may be related to a historical foraging strategy, used by the eagles before they spread to agricultural landscapes, since the current effects of body size strongly suggested food shortage in vole‐poor years.     

Histochemical observations on the crown skin of male baya: Lipids,lipase and phosphomonoesterases

Histochemical studies on localisation of lipids, lipase and phosphomonoeste-rases in the crown skin of male baya during non-breeding and breeding seasons were carried out. The results indicated a high turnover rate for lipid synthesis and its utilisation, increased acid and alkaline phosphatase activities in the crown skin of breeding male baya as compared to that in non-breeding season. It is surmised that crown skin of male baya becomes metabolically hyperactive during breeding season aiding processes such as cell growth and proliferation, keratinization and production of coloured feathers related to breeding.