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.     

Fifteen microsatellite loci characterized in the golden eagle Aquila chrysaetos (Accipitridae,Aves)

Polymorphic microsatellite loci were identified in order to study golden eagle (Aquila chrysaetos) population fragmentation. Twenty‐six published Aquila and eight published Haliaeetus microsatellite loci were tested for polymorphism in A. chrysaetos. Fifteen loci were polymorphic with between two and six alleles detected per locus. Observed heterozygosity ranged from 0.15 to 0.77 among 177 unrelated individuals from Scotland. There was no evidence for null alleles. Two pairs of loci (Hal‐10 & Aa15 and Hal‐10 & Aa26) displayed linkage disequilibrium.     

Fatal toxoplasmosis in a bald eagle (Haliaeetus leucocephalus)

Toxoplasma gondii tachyzoites were identified in the myocardium of a bald eagle (Haliaeetus leucocephalus) that died of necrotizing myocarditis. The diagnosis was confirmed by immunohistochemical staining with T. gondii-specific polyclonal antibodies. This is a new host record for T. gondii.     

Predator–prey dynamics of bald eagles and glaucous‐winged gulls at Protection Island,Washington, USA

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Long-term survival despite low genetic diversity in the critically endangered Madagascar fish-eagle

The critically endangered Madagascar fish-eagle ( Haliaeetus vociferoides ) is considered to be one of the rarest birds of prey globally and at significant risk of extinction. In the most recent census, only 222 adult individuals were recorded with an estimated total breeding population of no more than 100–120 pairs. Here, levels of Madagascar fish-eagle population genetic diversity based on 47 microsatellite loci were compared with its sister species, the African fish-eagle ( Haliaeetus vocifer ), and 16 of these loci were also characterized in the white-tailed eagle ( Haliaeetus albicilla ) and the bald eagle ( Haliaeetus leucocephalus ). Overall, extremely low genetic diversity was observed in the Madagascar fish-eagle compared to other surveyed Haliaeetus species. Determining whether this low diversity is the result of a recent bottleneck or a more historic event has important implications for their conservation. Using a Bayesian coalescent-based method, we show that Madagascar fish-eagles have maintained a small effective population size for hundreds to thousands of years and that its low level of neutral genetic diversity is not the result of a recent bottleneck. Therefore, efforts made to prevent Madagascar fish-eagle extinction should place high priority on maintenance of habitat requirements and reducing direct and indirect human persecution. Given the current rate of deforestation in Madagascar, we further recommend that the population be expanded to occupy a larger geographical distribution. This will help the population persist when exposed to stochastic factors (e.g. climate and disease) that may threaten a species consisting of only 200 adult individuals while inhabiting a rapidly changing landscape.     

Development and characterization of eleven microsatellite markers for an endangered cavefish (Triplophysa rosa Chen and Yang, 2005) using 454 sequencing

Triplophysa rosa is an endangered cave‐dwelling and endemic fish species found only in Chongqing, southwestern China. The genetic data available for this species is very limited. Polymorphic microsatellites were identified in the genome of T. rosa using 454 sequencing. Of the 145 loci screened, 106 were amplified successfully and 11 showed polymorphic patterns. The number of alleles ranged from 2 to 6, and the observed and expected heterozygosity varied from 0.061 to 0.543 (mean = 0.349) and from 0.248 to 0.789 (mean = 0.551), respectively. The polymorphic information content (PIC) ranged from 0.215 to 0.744 (mean = 0.486), indicating moderate levels of polymorphism. In addition, cross‐species amplification was tested for the 106 loci in Triplophysa moquensis, which showed a high level of transferability (76.4%), implying that the microsatellite markers developed here could be used effectively for other closely related species.     

Polymorphic microsatellite loci for paternity analysis in the Madagascar paradise flycatcher (Terpsiphone mutata: Aves)

Nine polymorphic microsatellite loci from the Madagascar paradise flycatcher Terpsiphone mutata were isolated using nonradioactive polymerase chain reaction (PCR)‐based techniques to screen an enriched genomic library. Seven polymorphic loci showed no evidence of null alleles and exhibited high levels of variation in 18 unrelated individuals (mean diversity = 0.80, mean number of alleles = 13.6). These loci are therefore suitable for analysis of population structure and paternity (exclusion probability for six unlinked loci = 0.9998).     

Isolation of polymorphic microsatellite loci in the clear lake gnat and two other phantom midge species of the genus Chaoborus (Diptera: Chaoboridae)

Chaoborus is of great interest to many freshwater ecologists. The adults can become pests in certain areas in North America and the larvae are an important food source for fish. In this preliminary study, we identified variable microsatellite loci in three species: Chaoborus astictopus (H_E = 0.52–0.76), Chaoborus americanus (H_E = 0.46–0.80) and Chaoborus punctipennis (H_E = 0.66–0.81). Using a biotin/streptavidin capture technique of repetitive sequences in a 96‐well format, we obtained microsatellite‐enriched genomic libraries for all three species and identified six polymorphic microsatellite markers for each species. None of the primers did yield a polymerase chain reaction fragment in a cross‐species test.     

Isolation and characterization of microsatellites in yellow perch (Perca flavescens)

A total of 45 microsatellite loci from yellow perch, Perca flavescens, were isolated and characterized. Among the 45 microsatellite loci, 32 had more than two alleles. A wild population of P. flavescens (n = 48) was used to examine the allele range of the microsatellite loci. Mendelian inheritance of alleles was confirmed by examining the amplified products in pair‐mated families. The number of alleles for the 32 polymorphic loci varied from two to 16, and observed heterozygosity ranged between 0.024 (YP79) and 0.979 (YP60). Cross‐species polymorphic amplification in four other Percidae species was successful for 22 loci.     

Polymorphic microsatellites in largemouth bronze gudgeon (Coreius guichenoti) developed from repeat‐enriched libraries and cross‐species amplifications

Largemouth bronze gudgeon (Coreius guichenoti) is a medium‐sized fish endemic from the upper Yangtze River of China and its survival is threatened by the construction of the Three Gorges Dam. This study reports 20 new polymorphic microsatellites from a repeat‐enriched genomic library with a mean number allele of 5.2, and observed and expected heterozygosities ranging from 0.035 to 1, and from 0.13 to 0.917, respectively. In a cross‐species amplification test, nine of the 37 tested loci were found to be also polymorphic in a congeneric species, brass gudgeon (C. heterodon). In addition, other four loci from common carp (Cyprinus carpio) were also polymorphic in C. guichenoti. Out of these 24 polymorphic microsatellites, only three loci significantly deviated from Hardy–Weinberg equilibrium in the sampled population (P < 0.0025), and all pairwise tests for linkage disequilibrium among loci were nonsignificant after applying sequential Bonferroni correction (P > 0.0026). These novel microsatellites provide sufficient levels of polymorphism for studies on population genetics and conservation in C. guichenoti and its related species.     

Development of microsatellite DNA loci from the wood stork (Aves,Ciconiidae, Mycteria americana)

We isolated 11 polymorphic microsatellite loci for wood stork (Mycteria americana). Polymerase chain reaction (PCR) primers and conditions are described for the amplification of five dinucleotide, one trinucleotide and five tetranucleotide microsatellite loci. The PCR primers were tested on two wood stork populations, Fazenda Ipiranga, Mato Grosso, Brazil (n = 11) and Tamiami West, Everglades, Florida, USA (n = 20). The primers yielded two to four alleles per locus, an observed heterozygosity of 0.0–0.727 and a polymorphic information content of 0.048–0.604. The low level of polymorphism for these markers is consistent with previous studies of this species.     

Isolation and characterization of microsatellite loci in the great bustard,Otis tarda

With the aim to study population genetics of the endangered great bustard, Otis tarda, dinucleotide microsatellite loci were isolated using an adapted hybrid‐capture enrichment protocol. This work reports the characterization of a set of six polymorphic microsatellite markers within the great bustard (n = 52). Results from cross‐species amplifications in several other members of the family Otididae demonstrate that five primer pairs also successfully amplified homologous loci outside the species Otis tarda.     

Isolation of microsatellites from two species of scleractinian coral

Microsatellites were isolated from two broadcast‐spawning species of scleractinian coral (Platygyra daedalea and Goniastrea favulus) from Australia's Great Barrier Reef. We found 27 microsatellites across both species, although only five loci were polymorphic in each species. Microsatellite loci displayed a wide range of diversity levels with four to 11 alleles per locus (H_O = 0.26–0.91) in P. daedalea and two to seven alleles per locus (H_O = 0.16–0.96) in G. favulus. Most loci showed departures from Hardy–Weinberg equilibrium which may reflect nonrandom mating but may also be related to difficulties associated with coral DNA.     

Characterization of 12 microsatellite primer pairs for the African grey parrot,Psittacus erithacus and their conservation across the Psittaciformes

This study describes 12 microsatellite loci identified in the African grey parrot Psittacus erithacus. Eleven were polymorphic, with observed heterozygosities 42–94% (average 68) and exclusion powers of P_E1 = 0.996 and P_E2 = 0.999. Microsatellites have previously been developed for a number of other parrots but showed limited cross‐species polymorphism. Here high levels of cross‐species amplification were observed: 71% of 32 Psittacines (22 genera). At least seven loci, 58%, were polymorphic in other African parrots as well as Neotropical and Australasian parrots, which diverged from the African parrots c30.6 and over 41.4 million years ago, respectively.     

Characterization,multiplex conditions,and cross‐species utility of tetranucleotide microsatellite loci for Blanchard's cricket frog (Acris crepitans blanchardi)

We have isolated and characterized 17 tetranucleotide microsatellite loci for Blanchard's cricket frog (Acris crepitans blanchardi), an anuran common in the central USA. Sixteen loci were organized into four multiplex amplification reactions. These loci were highly polymorphic when screened in 55 individuals from two distant populations, with 11–48 alleles per locus (average = 24.8). Observed and expected heterozygosities ranged from 0.18 to 0.97 and from 0.17 to 0.96, respectively. Nine loci were also polymorphic in Acris crepitans crepitans, with seven polymorphic in Acris gryllus. Five loci amplified in all three taxa. These loci will be useful for population‐ and species‐level investigations of this widespread group.     

Isolation and characterization of microsatellite markers in the southern emu‐wren (Stipiturus malachurus: Aves)

We isolated and characterized eight novel microsatellite loci in the southern emu‐wren (Stipiturus malachurus). We used nonradioactive polymerase chain reaction (PCR)‐based techniques to screen an enriched genomic DNA library. Based on genotypes from a single population, six loci showed no evidence of null alleles and were polymorphic (allele range = 2–9, mean heterozygosity = 0.57), and one locus was sex‐linked (N_A = 4). These loci were variable and had different allele size ranges in three other populations of southern emu‐wrens, and are therefore useful for determining levels of genetic diversity within and between populations of the species.     

Microsatellite markers from a commercially important South‐east Asian cyprinid,the silver barb (Barbodes gonionotus Bleeker)

Barbodes gonionotus (Puntius gonionotus/javanicus, Bleeker) is a cyprinid fish, endemic to South‐east Asia and is of great importance in both wild fisheries and in aquaculture in the region. It is a common to dominant species in the middle and lower stretches of medium and large rivers. Five highly polymorphic microsatellite loci are described (H_O = 0.7–0.9, number of alleles per locus = 10–37), which will be of use both in population genetic studies and for the assessment of true effective breeding population sizes in aquaculture stocks.     

Isolation and characterization of 13 polymorphic microsatellite loci for the Florida pompano,Trachinotus carolinus

Here we describe 13 polymorphic, dinucleotide microsatellite loci for Trachinotus carolinus (Florida pompano), isolated by using PIMA, a polymerase chain reaction‐based technique. The number of alleles at each locus ranged from 3 to 29 (mean = 11.4) in 45 specimens collected from bay and nearshore waters around St Petersburg, Florida. Levels of expected and observed heterozygosities ranged from 0.15 to 0.94 (mean = 0.69) and from 0.16 to 0.98 (mean = 0.70), respectively. No significant departures from Hardy–Weinberg equilibrium expectations were observed. In exact tests for genotypic disequilibrium, there was no evidence of linkage for any pair of loci. The ability of these markers to cross‐amplify in two congeneric Trachinotus species —T. falcatus (permit) and T. goodei (palometa) — was also assessed. The loci were well‐resolved, highly polymorphic, and independently segregating in these taxa, also suggesting a general utility for intraspecific studies, species identification, and investigation of interspecific hybridization.     

Movements by juvenile and immature Steller's Sea Eagles Haliaeetus pelagicus tracked by satellite

Twenty‐four juvenile Steller's Sea Eagles Haliaeetus pelagicus were tracked via satellite from natal areas in Magadan, Kabarovsk, Amur, Sakhalin and Kamchatka. Nestling dispersal occurred between 9 September and 6 December (n = 24), mostly 14 September–21 October, and did not differ among regions or years. Most eagles made stopovers of 4–28 days during migration. Migration occurred 9 September–18 January, mostly along previously described routes, taking 4–116 days to complete (n = 18). Eagles averaged 47.8 km/day excluding stopovers; 22.9 km/day including stopovers. The mean degrees of latitude spanned during migration was: Kamchatka, 2.1; Magadan, 11.6; Amur, 7.3; and Sakhalin, 1.1. Eagle winter range sizes varied. Eagles concentrated in 1–3 subareas within overall winter ranges. The mean size of the first wintering subareas was 274 km², the second 529 km², and the third 1181 km². Second wintering areas were south of first wintering areas. Spring migration started between 2 February and 31 March. Two eagles from Magadan were tracked onto summering grounds, well south of their natal areas. Both had early and late summering areas. One bird was followed for 25 months. It initiated its second autumn migration in the first half of October and arrived on its wintering grounds on 26 December. The second autumn migration covered 1839 km (20.9–22.4 km/day). Unlike its first winter when it used two subareas, this bird used only one subarea in 1998–99, but this was located near wintering areas used in 1997–98. It left its wintering ground between 13 April and 13 May, and arrived on its summering grounds between 7 June and 8 July. Unlike most satellite radiotracking studies, data are presented from a relatively large number of birds from across their breeding range, including new information on eagle movements on the wintering grounds and during the second year.