Mitochondrial phylogeography, subspecific taxonomy, and conservation genetics of sandhill cranes (Grus canadensis; Aves: Gruidae)

Six subspecies of sandhill cranes (Gruscanadensis) have been denoted based onperceived morphological and/or breedinglocality differences among them. Threesubspecies are migratory, breeding from thehigh arctic in North America and Siberia(lesser sandhill, G. c. canadensis),south through central Canada (Canadiansandhill, G. c. rowani) and into thenorthern United States (greater sandhill, G. c. tabida). A review of sandhill cranetaxonomy indicates that the size variation, onthe basis of which these subspecies were named,may be clinal and not diagnostic. The otherthree subspecies, all listed as endangered orthreatened, are non-migratory, resident inFlorida (G. c. pratensis), Mississippi(G. c. pulla), and Cuba (G. c.nesiotes). We used analysis of mitochondrialDNA control region (CR) sequences to determinewhether haplotypes representing currentsubspecies show any genetic cohesion or aremore consistent with a pattern of clinalvariation in morphology. CR sequences indicatethat only two highly divergent (5.3%) lineagesof sandhill cranes occur in North America: onelineage composed only of arctic-nesting G.c. canadensis, the other of the remainingNorth American subspecies (we lack data on theCuban population). The deep split betweenlineages is consistent with an estimatedisolation of approximately 1.5 Mya(mid-Pleistocene), while the distribution ofmutational changes within lineages isconsistent with an hypothesis of rapid,post-Pleistocene population expansions. Noother phylogeographic structuring is concordantwith subspecific boundaries, however, analysisof molecular variance indicates that there issignificant population genetic differentiationamong all subspecies except G. c. tabidaand G. c. rowani, which areindistinguishable. We suggest thatrecognition of the recently named G. c.rowani be abandoned.     

Genetic variation in the midcontinental population of sandhill cranes,Grus canadensis

Three subspecies of sandhill crane (Grus canadensis) are recognized in the Midcontinental population, the lesser (Grus c. canadensis), Canadian (G. c. rowani), and greater (G. c. tabida). Blood samples collected on the population's primary spring staging area in Nebraska, U.S.A., were used to resolve the genetic relationship among these subspecies. Phylogenetic analysis of 27 G. canadensis, by DNA sequencing of a 675 bp region of the mtDNA, supports the subspecies designations of G. c. canadensis and G. c. tabida. G. c. rowani individuals were intermediate with each of the other two subspecies. Genetic divergence ranged from 6.5 to 14.5% between G. c. canadensis and G. c. tabida, 0.5 to 6.6% within G. c. canadensis, and 0.1 to 6.0% within G. c. tabida. Sufficient DNA for analysis was obtained from shed feathers indicating a source of genetic material that does not require the capture or sacrifice of the birds. Other genetic markers and methods, including satellite telemetry, are required for obtaining detailed information on crane distributions as needed to establish effective management units for the MCP.     

Coccidia of whooping cranes

Coccidial oocysts were observed in 6 of 19 fecal samples from free-ranging whooping cranes (Grus americana) and 4 of 16 samples from captive whooping cranes. Eimeria gruis occurred in four free-ranging whooping cranes and E. reichenowi in two free-ranging and two captive whooping cranes. Fecal samples from two captive cranes contained oocysts of Isospora lacazei which was considered a spurious parasite. Oocysts of both species of Eimeria were prevalent in fecal samples collected from three free-ranging Canadian sandhill cranes (G. canadensis rowani) from whooping crane wintering grounds in Texas. These coccidia were prevalent also in fecal samples from 14 sandhill cranes (of 4 subspecies) maintained in captivity at the Patuxent Wildlife Research Center in Maryland.     

Weak large‐scale population genetic structure in a philopatric seabird,the European Shag Phalacrocorax aristotelis

Quantifying population genetic structure is fundamental to testing hypotheses regarding gene flow, population divergence and dynamics across large spatial scales. In species with highly mobile life‐history stages, where it is unclear whether such movements translate into effective dispersal among discrete philopatric breeding populations, this approach can be particularly effective. We used seven nuclear microsatellite loci and mitochondrial DNA (ND2) markers to quantify population genetic structure and variation across 20 populations (447 individuals) of one such species, the European Shag, spanning a large geographical range. Despite high breeding philopatry, rare cross‐sea movements and recognized subspecies, population genetic structure was weak across both microsatellites and mitochondrial markers. Furthermore, although isolation‐by‐distance was detected, microsatellite variation provided no evidence that open sea formed a complete barrier to effective dispersal. These data suggest that occasional long‐distance, cross‐sea movements translate into gene flow across a large spatial scale. Historical factors may also have shaped contemporary genetic structure: cluster analyses of microsatellite data identified three groups, comprising colonies at southern, mid‐ and northern latitudes, and similar structure was observed at mitochondrial loci. Only one private mitochondrial haplotype was found among subspecies, suggesting that this current taxonomic subdivision may not be mirrored by genetic isolation.     

Population and Harvest Dynamics of Midcontinent Sandhill Cranes

Sandhill cranes (Antigone canadensis) inhabiting the midcontinent of North America have been hunted since the 1960s under management goals of maintaining abundance, retaining geographic distribution, and maximizing sustainable harvest. Some biologists have raised concerns regarding harvest sustainability because sandhill cranes have lower reproductive rates than other game birds. We summarized demographic information in an age-structured matrix model to better understand population dynamics and harvest. Population indices and recovered harvest since the early 1980s suggest midcontinent sandhill cranes have experienced an average long-term annual growth of 0.9%; meanwhile, harvest has increased 1.8% annually. Adult survival and recruitment rates estimated from field data required modest adjustments (1–3%) so that model-derived growth rates matched growth estimated from a long-term survey (0.887 adult survival and 0.199 females/breeding female). Considering 0.9% long-term annual growth, sandhill cranes could be harvested at a rate of 6.6% if harvest was additive to natural mortality (assumed to be 0.05) or 11.3% if harvest mortality compensated for natural mortality. Life-history characteristics for long-lived organisms and demographic evidence suggested that hunter harvest was primarily additive. Differential harvest rates of segments of sandhill cranes in the midcontinent population derived from differential exposure to hunting suggested potentially unsustainable harvest for greater sandhill cranes (A. c. tabida) from 2 breeding segments. Overall, demographic evidence suggests that the harvest of sandhill cranes in the midcontinent population has been managed sustainably. Monitoring activities that reduce nuisance variation and estimate vital and harvest rates by subspecies would support continued management of sandhill cranes that are of interest to hunters and bird watchers. Published 2020. This article is a U.S. Government work and is in the public domain in the USA.     

Low genetic differentiation among reed warbler Acrocephalus scirpaceus populations across Europe

Migratory birds generally have higher dispersal propensity than resident species and are thus expected to show less genetic differentiation. On the other hand, specific migration patterns may promote genetic structure, such as in situations where migratory divides impede random mixing of individuals. Here we investigated population genetic structure and gene flow patterns in a polytypic passerine, the reed warbler Acrocephalus scirpaceus which shows a migratory divide in central Europe. Using ten polymorphic microsatellite loci and extensive sampling we found low but significant overall genetic differentiation (F_ST=0.013, G’_ST=0.078, D=0.063). Hierarchical F‐statistics and barrier analyses showed low but significant genetic differentiation of Iberian populations, and also slight genetic differences across the migratory divide and between subspecies (A. s. scirpaceus and A. s. fuscus). Three individual‐based Bayesian methods, however, inferred a single genetic unit. Our study thus found low levels of genetic differentiation among reed warbler populations but this genetic differentiation was not pronounced enough to detect a clear population structure using the microsatellite data and no prior information on geographic location of the sampled individuals. This result indicates high levels of gene flow and suggests a possibly recent divergence of European populations after a rapid range expansion. Further studies are necessary to assess divergence times and to reveal the evolutionary history of the reed warbler populations.     

Flyway structure in the circumpolar greater white‐fronted goose

Dispersal and migratory behavior are influential factors in determining how genetic diversity is distributed across the landscape. In migratory species, genetic structure can be promoted via several mechanisms including fidelity to distinct migratory routes. Particularly within North America, waterfowl management units have been delineated according to distinct longitudinal migratory flyways supported by banding data and other direct evidence. The greater white‐fronted goose (Anser albifrons) is a migratory waterfowl species with a largely circumpolar distribution consisting of up to six subspecies roughly corresponding to phenotypic variation. We examined the rangewide population genetic structure of greater white‐fronted geese using mtDNA control region sequence data and microsatellite loci from 23 locales across North America and Eurasia. We found significant differentiation in mtDNA between sampling locales with flyway delineation explaining a significant portion of the observed genetic variation (~12%). This is concordant with band recovery data which shows little interflyway or intercontinental movements. However, microsatellite loci revealed little genetic structure suggesting a panmictic population across most of the Arctic. As with many high‐latitude species, Beringia appears to have played a role in the diversification of this species. A common Beringian origin of North America and Asian populations and a recent divergence could at least partly explain the general lack of structure at nuclear markers. Further, our results do not provide strong support for the various taxonomic proposals for this species except for supporting the distinctness of two isolated breeding populations within Cook Inlet, Alaska (A. a. elgasi) and Greenland (A. a. flavirostris), consistent with their subspecies status.     

Pulmonary lesions in disseminated visceral coccidiosis of sandhill and whooping cranes

Fifty cranes, consisting of 46 sandhill (Grus canadensis) and four whooping cranes (Grus americana), were studied. Eighteen sandhill cranes and the four whooping cranes were naturally infected with disseminated visceral coccidiosis (DVC). The remaining sandhill cranes were chicks experimentally infected with oocysts of Eimeria reichenowi and/or E. gruis; five chicks served as controls. There were no clinical signs attributed to respiratory infection. Necropsy of naturally infected adult birds revealed nodules in many organs, including the lung, air sacs, trachea and nares. Artificially infected sandhill cranes and the whooping crane chicks that died from DVC had congestion and consolidated areas in the lung with frothy fluid in the airways. Grossly visible nodules were observed from 10 days postinoculation. Granulomatous pneumonia and tracheitis were observed with light microscopy. Lesions were associated with merogonic and gametogonic stages of eimerian coccidia. Granulomas and granulomatous foci contained parasitized large mononuclear cells. Merogonic stages were seen in lymphoid cells by ultrastructural examination. Oocysts were observed in the trachea and bronchial mucosa and admixed with exudate in the airways, indicating that crane eimerians can complete their life cycle at these sites. Of the few eimeriid coccidia that have extraintestinal stages of development in birds and mammals, only the species in cranes complete their life cycle in both the digestive and respiratory tracts.     

Population genetics and morphological comparisons of migratory European (Hirundo rustica rustica) and sedentary East-Mediterranean (Hirundo rustica transitiva) barn swallows

Speciation processes are largely determined by the relative strength of divergent selection versus the magnitude of gene flow. The barn swallow (Hirundo rustica) has a broad geographic distribution that encompasses substantial geographic variation in morphology and behavior. The European (H. r. rustica) and East-Mediterranean (H. r. transitiva) subspecies are closely related, despite differing in morphological and life-history traits. To explore patterns of genetic differentiation and gene flow, we compared morphological and genetic variation among the nonmigratory breeding population of H. r. transitiva from Israel and the migratory population of H. r. rustica that passes through Israel and compared it with the genetic differentiation between H. r. transitiva from Israel and a breeding population of H. r. rustica from the United Kingdom that uses a different migratory flyway. Mitochondrial haplotype network analysis suggests that the European and East-Mediterranean populations are intermixed, although there was low but significant genetic differentiation between the subspecies based on both mitochondrial (F(ST) = 0.025-0.033) and microsatellite (F(ST) = 0.009-0.014) loci. Coalescent-based analyses suggest recent divergence and substantial gene flow between these populations despite their differences in morphological and behavioral traits. The results suggest that these subspecies are undergoing a differentiation process in the face of gene flow, with selection possibly operating on sexually selected traits.     

Genetic structure of muskrat (Ondatra zibethicus) and its concordance with taxonomy in North America

Extrinsic factors such as physical barriers play an important role in shaping population genetic structure. A reduction in gene flow leading to population structuring may ultimately lead to population divergence. These divergent populations are often considered subspecies. Because genetic differentiation may represent differences between subspecies, patterns of genetic structure should reflect subspecies groupings. In this study, we examine the contemporary population genetic structure of muskrat (n = 331) and assess the relevance of 4 geographically distinct subspecies designations across northern North America using 9 microsatellite loci. We predicted that patterns of gene flow and genetic structure would reflect the described subspecies. We found evidence of genetic differentiation between western and eastern regions, and muskrats from Newfoundland (NF) showed significantly lower genetic diversity than central regions. A strong isolation by distance pattern was also detected within the eastern cluster. Our results did not differentiate Ondatra zibethicus spatulus (northwest) from O. z. albus (central), but they suggest a distinction between O. z. obscurus (NF) and O. z. zibethicus (east). This study highlights the need for more phylogenetic studies in order to better understand intraspecific divergence and the genetic characterization of subspecies.     

THE HISTORICAL PATTERN OF GENE FLOW AMONG MIGRATORY AND NONMIGRATORY POPULATIONS OF PRAIRIE WARBLERS (AVES: PARULINAE)

Within a group of interbreeding organisms, the balance of gene flow among populations and microevolutionary forces acting within populations is expected to result in clinal transitions in the phenotypes possessed by members of differentiated populations. Discontinuous variation between geographically adjacent populations suggests the presence of a significant barrier to gene flow. Here I present genetic evidence for restricted gene flow between migratory and nonmigratory populations of prairie warblers. The nonmigratory form of this species is restricted to coastal mangroves in Florida and is morphologically distinguishable from the typical, migratory form that occurs across the remainder of the eastern United States. Pairs of migratory populations exhibited little population subdivision (Φ_ST ? 0.09), whereas pairs of migratory and nonmigratory populations are much more differentiated (Φ_ST = 0.27–0.42). A phylogenetic analysis of mitochondrial DNA haplotypes did not offer evidence of long-term isolation of migratory and nonmigratory populations. Together with the population genetic analysis, the phylogenetic relationship of haplotypes suggests that isolation between these forms must have arisen relatively recently in their history. Evidence for significant population structure is unexpected, given the geographic proximity of migratory and nonmigratory populations, the capacity for long-distance movements (e.g., migration) by prairie warblers, and several previous studies of population structure in North American birds. However, the findings are consistent with the geographic distribution of morphological and behavioral variation and demonstrate that significant boundaries between populations of vagile organisms may be relatively cryptic.     

Range expansion and the possibility of an emerging contact zone between two subspecies of Chiffchaff Phylloscopus collybita ssp.

The Chiffchaff Phylloscopus collybita is represented in Sweden by two different subspecies; the northern well-established abietinus and the southern recently established collybita which has expanded its range northward during the past two decades. At present, an area approximately 500 km wide separates the two subspecies. In order to document differences between the northern and southern populations we compared morphology, vocalisation, habitat choice, and neutral genetic variation in mitochondrial (mt) DNA and at four microsatellite loci of 30 male Chiffchaffs from each subspecies. Our results show significant differences in several morphological traits and in song. Playback experiments revealed a significant difference in aggressive behaviour depending on which population-specific song that was played to the birds. Mitochondrial DNA was geographically structured with ∼90% of the birds carrying a mt haplotype matching their sample population. No allelic differences at the microsatellite loci were found between populations. Our data demonstrate a substantial differentiation between the northern and southern populations despite gene flow, clearly separating them into the subspecies abietinus and collybita .     

A population genetic comparison of argali sheep (Ovis ammon) in Mongolia using the ND5 gene of mitochondrial DNA; implications for conservation

We sequenced 556 bp of the mitochondrial ND5 gene to infer aspects of population structure and to test subspecific designations of argali sheep (Ovis ammon) in Mongolia. Analysis of molecular variance (AMOVA) revealed greater variation within than among putative subspecies and populations, suggesting high levels female-mediated gene flow. Compared with bighorn sheep (O. canadensis) in North America, substantially less differentiation in mitochondrial DNA was found among argali populations over 1200 km than was found among bighorn populations over 250 km. This result is consistent with differences in argali and bighorn life history traits. Argali run for long distances across open terrain in the presence of a threat rather than running up into steep escape terrain like bighorn sheep do. Our results suggest recognizing only one Evolutionary Significant Unit (subspecies) of argali in Mongolia, but they may support recognizing two Management Units, because two regions do exhibit slightly different haplotype frequencies at the ND5 gene of mtDNA.     

Fusarium mycotoxins from peanuts suspected as a cause of sandhill crane mortality

An estimated 9,500 sandhill cranes (Grus canadensis) died in Gaines County, Texas and Roosevelt County, New Mexico between 1982 and 1987. The predominant clinical sign observed in sick cranes was their inability to hold their heads erect, both while standing and flying. Multiple muscle hemorrhages and submandibular edema were the most common lesions seen at necropsy. Mycotoxins produced by Fusarium sp. growing during cold, wet weather on peanuts left in the field after harvest, the predominant foods of the dead cranes at the time of these mortality events, were identified as the most likely cause of this mortality. Rendering moldy peanuts inaccessible to the cranes by conventional tillage resulted in reduced crane mortality in these areas.     

Breeding site fidelity and winter admixture in a long-distance migrant, the tufted duck (Aythya fuligula)

Long-distance migrants are, by definition, highly mobile but it is poorly understood if this leads to high rates of gene flow and an essentially panmictic global population structure. Genetic divergence in migratory species could be promoted, for example, by fidelity to distinct migratory pathways. In this study, we investigate the population genetic structure of tufted duck (Aythya fuligula), a long-distance migrant with a largely continuous breeding distribution across Eurasia. Distinct, longitudinally oriented flyways have been postulated based on geographically disjunct wintering areas and are supported by evidence from ringing data. We generated sequences of the mitochondrial control region and multi-locus microsatellite genotypes for several hundreds of samples from the European and Asian breeding and wintering grounds including some individuals infected with highly pathogenic avian influenza virus H5N1. Significant differentiation between breeding sites was observed for both marker types, but F(ST) values were approximately 10 times higher for maternally inherited mitochondrial DNA than for biparentally transmitted nuclear markers. The genetic differentiation between the postulated European and Asian flyways was similar to that observed within continents and, in general, genetic divergence was not associated with geographic distance. Neither marker type showed evidence of genetic substructure among aggregations on the European wintering grounds. Our results suggest some breeding site fidelity, especially in females, but extensive population admixture on the wintering grounds. Several scenarios may explain the observed lack of genetic divergence between Europe and Asia including non-equilibrium conditions following a recent range expansion or contemporary gene flow across the postulated migratory divides.     

Fecal corticosterone reflects serum corticosterone in Florida sandhill cranes

Florida sandhill cranes (Grus canadensis pratensis) were conditioned to confinement 6 hr/day for 7 days. On day 8, each bird's jugular vein was catheterized, blood samples were drawn, and each crane was confined for 6 hr. Using a randomized, restricted cross-over design, cranes were injected intravenously with either 0.9% NaCl solution or ACTH (cosyntropin; Cortrosyn; 0.25 mg). During the 6 hr of confinement, fecal samples (feces and urine) were collected from each of five cranes immediately after defecation. Individual fecal samples were collected approximately at hourly intervals and assayed for corticosterone. We showed previously that serum corticosterone did not vary significantly following saline injection, but peaked significantly 60 min after ACTH injection. Maximal fecal corticosterone concentrations (ng/g) were greater (P < 0.10; median 1087 ng/g) following ACTH stimulation compared to maximal fecal corticosterone concentrations at the end of acclimation (day 7; median 176) and following saline treatment (median 541). In cranes under controlled conditions, fecal corticosterone concentration reflects serum corticosterone levels, fecal corticosterone, Grus canadensis pratensis, sandhill cranes, serum corticosterone levels.     

Migratory birds of central Washington as reservoirs of Campylobacter jejuni

Migratory ducks, Canada geese, and sandhill crane from the Pacific North American Flyway have been screened for Campylobacter spp. Samples (298) from these birds were examined and the incidence of Campylobacter spp. in the samples were as follows: sandhill crane (Grus canadensis tabida), 81%; ducks (Aythya collaris, Anas carolinensis, Aythya americana, and Anas platyrhynchos), 73%; and Canada geese (Branta canadensis), 5%. All isolates were identified as Campylobacter jejuni. To our knowledge this is the first report of the isolation of C. jejuni from sandhill crane. The high frequency of isolation in both the sandhill crane and migratory ducks indicated that these bird populations may play a significant role in the dissemination of the bacterium. Because of their migratory habits, these birds may be particularly important in spreading C. jejuni to remote areas.     

Avian pox in Florida sandhill cranes.

Cutaneous elevations were present on the feet, legs and heads of four Florida sandhill cranes, Grus canadensis pratensis, (one free-living, three penreared birds). As a result of examination of the elevations by light- and electron microscopy, it was determined that the lesions were caused by poxvirus. This is the first record of pox in cranes in North America.     

Capture of sandhill cranes using alpha-chloralose

From 1990-2001, we made 188 successful captures of 166 different greater sandhill cranes (Grus canadensis tabida) through experimental use of alpha-chloralose (AC). Most captures took place during September (72.3%; n = 136), followed by August (14.9%; n = 28), and October (12.8%; n = 24). Territorial pairs were captured more successfully than family groups. Overall morbidity (6.4%) and mortality (4.3%) were lower than most other capture techniques for sandhill cranes. Exertional myopathy (EM) was the most common complication observed using AC (3.7%). Sedation level (chi5(2) = 25.9, P < 0.01) and month of capture (chi2(2) = 12.3, P < 0.01) were both associated with the presence of EM in cranes captured with AC. A logistic regression model suggests lighter sedation and the months of August and October are potential risk factors for EM in sandhill cranes captured with AC in Wisconsin (USA).     

Hybridization between subspecies of waterbuck (Kobus ellipsiprymnus) in zones of overlap with limited introgression

Two subspecies of waterbuck (Kobus ellipsiprymnus), common (Kobus ellipsiprymnus ellipsiprymnus) and defassa (Kobus ellipsiprymnus defassa), are recognized based on differences in rump pattern, coat colour and geographical distribution. These forms are parapatrically distributed with an area of range overlap in East Africa, where phenotypically intermediate populations occur. Variation in 478 bp of the mitochondrial DNA control region and 14 polymorphic microsatellite loci were used to describe the genetic structure and phylogeographical pattern of the species, and to assess if the intermediate populations are the results of hybridization. In total, 186 individuals from 11 localities were analysed. A high degree of genetic differentiation was found between subspecies, although this was most evident from the microsatellite data. Hybridization was suggested in the phenotypically and geographically intermediate Nairobi NP population in Kenya. A neighbour-joining (NJ) tree based on microsatellite population genetic distances grouped Nairobi between the common and defassa populations, and a Bayesian analysis clearly showed introgression. Individuals sampled in Samburu NP, Kenya, had a common waterbuck phenotype, but introgression was suggested by both markers. Although a high degree of maternal defassa input was indicated from the sequence data, the Samburu population grouped with the common waterbuck in the microsatellite population genetic distance tree, with high support. Analyses of linkage disequilibrium and maximum-likelihood estimates of genetic drift suggested that admixture between subspecies is a recent event. The fact that introgression is limited between subspecies could be caused by chromosomal differences, hindering gene flow between common and defassa waterbuck.