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Gopherus agassizii : Desert tortoise

The desert tortoise is one of four allopatric North American tortoises. It occurs in the Mojave and Sonoran deserts of the southwestern United States and Mexico. Auffenberg (1976) divided the genus Gopherus (consisting of four species, G. agassizi , G. berlandieri , G.flavomarginatus , and G. polyphemus ) in two osteological groups. Bramble (1982), using morphological and palaeontological data, divided the genus Gopherus into two separate complexes, each with two species. He established a new genus, Scaptochelys , for agassizi and berlandieri , retaining Gopherus for polyphemus and flavomarginatus . Bour and Dubois (1984) noted that Xerobates Agassiz had priority over Scaptochelys Bramble. Using mitochondrial DNA (mtDNA), Lamb et al. (1989) evaluated the evolutionary relationships of the North American tortoises, particularly the desert tortoise. They concluded that the mtDNA analysis provides strong support for generic recognition of the two distinct species groups described by Bramble (1982). Until a few decades ago, the desert tortoise was widespread at lower elevations throughout the Mojave and Sonoran deserts of the U.S.A. In the northern and western parts of the geographic range, large and relatively homogeneous populations with densities exceeding 1,000/sq km extended throughout parts of California, and probably into Nevada and Utah. In terms of biomass, the tortoise played an important role in the ecosystems. In most areas, numbers have declined dramatically and the extent of populations has been reduced. Most populations are now isolated and low in numbers. Conservation of the desert tortoise is a highly visible and political issue in the U.S.A., but not in Mexico.

Book chapter

Intraspecific evolutionary relationships among peregrine falcons in western North American high latitudes

Subspecies relationships within the peregrine falcon ( Falco peregrinus ) have been long debated because of the polytypic nature of melanin-based plumage characteristics used in subspecies designations and potential differentiation of local subpopulations due to philopatry. In North America, understanding the evolutionary relationships among subspecies may have been further complicated by the introduction of captive bred peregrines originating from non-native stock, as part of recovery efforts associated with mid 20 th century population declines resulting from organochloride pollution. Alaska hosts all three nominal subspecies of North American peregrine falcons– F . p . tundrius , anatum , and pealei –for which distributions in Alaska are broadly associated with nesting locales within Arctic, boreal, and south coastal maritime habitats, respectively. Unlike elsewhere, populations of peregrine falcon in Alaska were not augmented by captive-bred birds during the late 20 th century recovery efforts. Population genetic differentiation analyses of peregrine populations in Alaska, based on sequence data from the mitochondrial DNA control region and fragment data from microsatellite loci, failed to uncover genetic distinction between populations of peregrines occupying Arctic and boreal Alaskan locales. However, the maritime subspecies, pealei , was genetically differentiated from Arctic and boreal populations, and substructured into eastern and western populations. Levels of interpopulational gene flow between anatum and tundrius were generally higher than between pealei and either anatum or tundrius . Estimates based on both marker types revealed gene flow between augmented Canadian populations and unaugmented Alaskan populations. While we make no attempt at formal taxonomic revision, our data suggest that peregrine falcons occupying habitats in Alaska and the North Pacific coast of North America belong to two distinct regional groupings–a coastal grouping ( pealei ) and a boreal/Arctic grouping (currently anatum and tundrius )–each comprised of discrete populations that are variously intra-regionally connected.

Alaska

Intercontinental genetic structure and gene flow in Dunlin (Calidris alpina), a potential vector of avian influenza

Waterfowl (Anseriformes) and shorebirds (Charadriiformes) are the most common wild vectors of influenza A viruses. Due to their migratory behavior, some may transmit disease over long distances. Migratory connectivity studies can link breeding and nonbreeding grounds while illustrating potential interactions among populations that may spread diseases. We investigated Dunlin ( Calidris alpina ), a shorebird with a subspecies ( C. a. arcticola ) that migrates from nonbreeding areas endemic to avian influenza in eastern Asia to breeding grounds in northern Alaska. Using microsatellites and mitochondrial DNA, we illustrate genetic structure among six subspecies: C. a. arcticola , C. a. pacifica , C. a. hudsonia , C. a. sakhalina , C. a. kistchinski , and C. a. actites . We demonstrate that mitochondrial DNA can help distinguish C. a. arcticola on the Asian nonbreeding grounds with >70% accuracy depending on their relative abundance, indicating that genetics can help determine whether C. a. arcticola occurs where they may be exposed to highly pathogenic avian influenza (HPAI) during outbreaks. Our data reveal asymmetric intercontinental gene flow, with some C. a. arcticola short-stopping migration to breed with C. a. pacifica in western Alaska. Because C. a. pacifica migrates along the Pacific Coast of North America, interactions between these subspecies and other taxa provide route for transmission of HPAI into other parts of North America.

Evolutionary Applications

A genetic study to aid in restoration of murres, guillemots and murrelets to the Gulf of Alaska

Genetic data are needed to aid in restoring several species of seabirds to the Gulf of Alaska. We analyzed sequence variation in mitochondrial DNA, microsatellite DNA and nuclear introns in samples of commom murres ( Uria aalge ), pigeon guillemots ( Cepphus columba ) and marbled murrelets ( Brachyramphus marmoratus ) from throughout the North Pacific. Data were analyzed using traditional approaches, nested clade analyses and assignment tests. No cryptic species were found, and there was no strong evidence for inbreeding, low genetic variation, or souce or sink regions in any them. Pacific common murres constitute a single genetic management unit (MU), but hybridization occurs between common and thick-billed murres ( U. lomvia ). In contrast, gene flow in pigeon guillemots is very restricted and population genetic structure is very strong; guillemots from the spill area are part of a MU that extends from the Alaska Peninsula to somewhere between Prince William Sound and Vancouver Island. Marbled murrelets in the spill area are part of a MU that extends from the Alaska Peninsula to at least British Columbia; tree- and ground-nesting murrelets are not genetically differentiated. Little if any hybridization occurs between marbled and Kittlitz's murrelets.

Alaska

Environmental DNA assays for invasive populations of the Black Carp, Mylopharyngodon piceus, in North America

The Black Carp, Mylopharyngodon piceus, is an increasingly widespread invasive species in North America that threatens freshwater mussel populations. We developed four qPCR assays for detecting environmental DNA (eDNA) from these Black Carp populations. Assays were designed to target four mitochondrial DNA loci and were based on 34 complete mitochondrial genome sequences, including 29 generated in this study from samples obtained in three countries. Assays were validated for taxon specificity with in silico comparisons against archived DNA sequences and with in vitro tests of 41 DNA samples from Black Carp, as well as DNA samples from 30 non‐target fish species, all from the Mississippi River Basin. All four assays were able to detect the DNA of all Black Carp samples and did not exhibit any positive results with DNA from other tested species. Tests conducted in round‐robin fashion among three different laboratories found that all four assays were able to detect DNA at very low template concentrations (limits of detection = 3 copies/qPCR, limits of quantification = 16‐64 copies/qPCR) and, as part of in situ validation, were successful in detecting eDNA from Black Carp in aquaculture ponds. Despite some challenges with other attempts at in situ validation, the assays were also effective in detecting Black Carp eDNA in water samples from a drainage ditch in the upper reaches of the species’ range that was known to contain juvenile Black Carp, as well as in water samples from the Missisippi River and a connected oxbow lake in the lower reaches of the species range.

Transactions of the American Fisheries Society

The importance of genetic verification for determination of Atlantic salmon in north Pacific waters

Genetic analyses of two unknown but putative Atlantic salmon Salmo salar captured in the Copper River drainage, Alaska, demonstrated the need for validation of morphologically unusual fishes. Mitochondrial DNA sequences (control region and cytochrome b) and data from two nuclear genes [first internal transcribed spacer (ITS-1) sequence and growth hormone (GH1) amplification product] indicated that the fish caught in fresh water on the Martin River was a coho salmon Oncorhynchus kisutch, while the other fish caught in the intertidal zone of the Copper River delta near Grass Island was an Atlantic salmon. Determination of unusual or cryptic fish based on limited physical characteristics and expected seasonal spawning run timing will add to the controversy over farmed Atlantic salmon and their potential effects on native Pacific species. It is clear that determination of all putative collections of Atlantic salmon found in Pacific waters requires validation. Due to uncertainty of fish identification in the field using plastic morphometric characters, it is recommended that genetic analyses be part of the validation process. ?? 2003 The Fisheries Society of the British Isles.

Journal of Fish Biology

Population status and population genetics of northern leopard frogs in Arizona

Increasing isolation of populations by habitat fragmentation threatens the persistence of many species, both from stochastic loss of small isolated populations, and from inbreeding effects in populations that have become genetically isolated. In the southwestern United States, amphibian habitat is naturally patchy in occurrence because of the prevailing aridity of the region. Streams, rivers, and other wetlands are important both as habitat and as corridors that connect populations. However, populations of some species have become more fragmented and isolated by habitat degradation and loss. Northern leopard frogs (Rana pipiens) have experienced serious declines in the Southwest. We conducted an extensive survey across the known range of northern leopard frogs in Arizona to determine the current distribution and abundance of the species. From a range that once spanned much of the northern and central part of the State, northern leopard frogs have been reduced to three or four widely separated populations, near Lyman Lake in east-central Arizona, in the Stoneman Lake area south of Flagstaff, along Truxton Wash near Peach Springs, and a population of uncertain extent on Navajo Nation lands. The Lyman Lake and Truxton Wash populations are small and extremely isolated. The Stoneman Lake population, however, is an extensive metapopulation spread across several stream drainages, including numerous ponds, wetlands, and artificial tanks. This is the only population in Arizona that is increasing in extent and numbers, but there is concern about the apparent introduction of nonnative genetic stock from eastern North America into this area. We analyzed genetic diversity within and genetic divergence among populations of northern leopard frogs, across both extant and recently extirpated populations in Arizona. We also analyzed mitochondrial DNA to place these populations into a larger phylogenetic framework and to determine whether any populations contained genetic material not native to the region. We found a high level of genetic divergence among the population centers (Lyman Lake, Stoneman Lake, Truxton Wash), and low genetic diversity in the small populations at Lyman Lake and Truxton. The extensive population in the Stoneman Lake area had high genetic diversity and relatively high gene flow among ponds and tanks across the entire extent of the area. However, this population also contained a mitochondrial haplotype from northern leopard frogs from the northeastern United States or southeastern Canada, probably representing the introduction of released pets or laboratory animals. These eastern frogs were extensively distributed through this population, and probably contributed to its high genetic diversity. Genetic diversity in the outlying populations such as Truxton Wash, East Buckskin Tank, and Hess Tank was low and showed signs of recent bottlenecks. However, supplementing genetic diversity in these native populations with artificial gene flow from the Stoneman Lake area may only be advisable in extreme situations for which there are no other alternatives. Until the nature and effects of genetic mixing of eastern and western genetic stocks of northern leopard frogs are better understood, the long-term persistence of the species in the Southwest may be best served by retaining as much genetic integrity of remaining native populations as possible.

Arizona

Hierarchical spatial genetic structure in a distinct population segment of greater sage-grouse

Greater sage-grouse ( Centrocercus urophasianus ) within the Bi-State Management Zone (area along the border between Nevada and California) are geographically isolated on the southwestern edge of the species’ range. Previous research demonstrated that this population is genetically unique, with a high proportion of unique mitochondrial DNA (mtDNA) haplotypes and with significant differences in microsatellite allele frequencies compared to populations across the species’ range. As a result, this population was considered a distinct population segment (DPS) and was recently proposed for listing as threatened under the U.S. Endangered Species Act. A more comprehensive understanding of the boundaries of this genetically unique population (where the Bi-State population begins) and an examination of genetic structure within the Bi-State is needed to help guide effective management decisions. We collected DNA from eight sampling locales within the Bi-State (N = 181) and compared those samples to previously collected DNA from the two most proximal populations outside of the Bi-State DPS, generating mtDNA sequence data and amplifying 15 nuclear microsatellites. Both mtDNA and microsatellite analyses support the idea that the Bi-State DPS represents a genetically unique population, which has likely been separated for thousands of years. Seven mtDNA haplotypes were found exclusively in the Bi-State population and represented 73 % of individuals, while three haplotypes were shared with neighboring populations. In the microsatellite analyses both STRUCTURE and FCA separate the Bi-State from the neighboring populations. We also found genetic structure within the Bi-State as both types of data revealed differences between the northern and southern part of the Bi-State and there was evidence of isolation-by-distance. STRUCTURE revealed three subpopulations within the Bi-State consisting of the northern Pine Nut Mountains (PNa), mid Bi-State, and White Mountains (WM) following a north–south gradient. This genetic subdivision within the Bi-State is likely the result of habitat loss and fragmentation that has been exacerbated by recent human activities and the encroachment of singleleaf pinyon ( Pinus monophylla ) and juniper ( Juniperus spp.) trees. While genetic concerns may be only one of many priorities for the conservation and management of the Bi-State greater sage-grouse, we believe that they warrant attention along with other issues (e.g., quality of sagebrush habitat, preventing future loss of habitat). Management actions that promote genetic connectivity, especially with respect to WM and PNa, may be critical to the long-term viability of the Bi-State DPS.

California, Nevada

Diversity and distribution of white-tailed deer mtDNA lineages in chronic wasting disease (CWD) outbreak areas in southern Wisconsin, USA

Chronic wasting disease (CWD) is a transmissible spongiform encephalopathy affecting North American cervids. Because it is uniformly fatal, the disease is a major concern in the management of white-tailed deer populations. Management programs to control CWD require improved knowledge of deer interaction, movement, and population connectivity that could influence disease transmission and spread. Genetic methods were employed to evaluate connectivity among populations in the CWD management zone of southern Wisconsin. A 576-base-pair region of the mitochondrial DNA of 359 white-tailed deer from 12 sample populations was analyzed. Fifty-eight variable sites were detected within the sequence, defining 43 haplotypes. While most sample populations displayed similar levels of haplotype diversity, individual haplotypes were clustered on the landscape. Spatial clusters of different haplotypes were apparent in distinct ecoregions surrounding CWD outbreak areas. The spatial distribution of mtDNA haplotypes suggests that clustering of the deer matrilineal groups and population connectivity are associated with broad-scale geographic landscape features. These landscape characteristics may also influence the contact rates between groups and therefore the potential spread of CWD; this may be especially true of local disease spread between female social groups. Our results suggest that optimal CWD management needs to be tailored to fit gender-specific dispersal behaviors and regional differences in deer population connectivity. This information will help wildlife managers design surveillance and monitoring efforts based on population interactions and potential deer movement among CWD-affected and unaffected areas.

Wisconsin

Geographic distribution of the mid-continent population of sandhill cranes and related management applications

The Mid-continent Population (MCP) of sandhill cranes ( Grus canadensis ) is widely hunted in North America and is separated into the Gulf Coast Subpopulation and Western Subpopulation for management purposes. Effective harvest management of the MCP requires detailed knowledge of breeding distribution of subspecies and subpopulations, chronology of their use of fall staging areas and wintering grounds, and exposure to and harvest from hunting. To address these information needs, we tagged 153 sandhill cranes with Platform Transmitting Terminals (PTTs) during 22 February–12 April 1998–2003 in the Central and North Platte River valleys of south-central Nebraska. We monitored PTT-tagged sandhill cranes, hereafter tagged cranes, from their arrival to departure from breeding grounds, during their fall migration, and throughout winter using the Argos satellite tracking system. The tracking effort yielded 74,041 useable locations over 49,350 tag days; median duration of tracking of individual cranes was 352 days and 73 cranes were tracked >12 months. Genetic sequencing of mitochondrial DNA (mtDNA) from blood samples taken from each of our random sample of tagged cranes indicated 64% were G. c. canadensis and 34% were Grus canadensis tabida . Tagged cranes during the breeding season settled in northern temperate, subarctic, and arctic North America (U.S. [23%, n = 35], Canada [57%, n = 87]) and arctic regions of northeast Asia (Russia [20%, n = 31]). Distribution of tagged cranes by breeding affiliation was as follows: Western Alaska–Siberia (WA–S, 42 ± 4% [SE]), northern Canada–Nunavut (NC–N, 21 ± 4%), west-central Canada–Alaska (WC–A, 23 ± 4%) and East-central Canada–Minnesota (EC–M, 14 ± 3%). All tagged cranes returned to the same breeding affiliation used during the previous year with a median distance of 1.60 km (range: 0.08–7.7 km, n = 53) separating sites used in year 1 and year 2. Fall staging occurred primarily in central and western Saskatchewan (69%), North Dakota (16%), southwestern Manitoba (10%), and northwestern Minnesota (3%). Space-use sharing indices showed that except for NC–N and WC–A birds, probability of finding a crane from one breeding affiliation within the home range of another breeding affiliation was low during fall staging. Tagged cranes from WC–A and EC–M breeding affiliations, on average, spent 25 and 20 days, respectively, longer on fall staging areas in the northern plains than did WA–S and NC–N birds. Cranes in the NC–N, WA–S, and WC–A affiliations spent 99%, 74%, and 64%, respectively, of winter in western Texas in Hunting Zone A; EC–M cranes spent 83% of winter along the Texas Gulf Coast in Hunting Zone C. Tagged cranes that settled within the breeding range of the Gulf Coast Subpopulation spent 28% and 42% of fall staging and winter within the range of the Western Subpopulation, indicating sufficient exchange of birds to potentially limit effectiveness of MCP harvest management. Harvests of EC–M and WC–A cranes during 1998–2003 were disproportionately high to their estimated numbers in the MCP, suggesting more conservative harvest strategies may be required for these subpopulations in the future, and for sandhill cranes to occupy major parts of their historical breeding range in the Prairie Pothole Region. Exceptionally high philopatry of MCP cranes of all 4 subpopulations to breeding sites coupled with strong linkages between crane breeding distribution, and fall staging areas and wintering grounds, provide managers guidance for targeting MCP crane harvest to meet management goals. Sufficient temporal or spatial separation exists among the 4 subpopulations on fall staging areas and wintering grounds to allow harvest to be targeted at the subpopulation level in all states and provinces (and most hunting zones within states and provinces) when conditions warrant. Knowledge gained from our study provides decision-makers in the United States, Canada, Mexico, and Russia with improved guidance for developing sound harvest regulations, focusing conservation efforts, and generating collaborative efforts among these nations on sandhill crane research and management to meet mutually important goals.

Nebraska

Effects of Climate and land use on diversity, prevalence, and seasonal transmission of avian hematozoa in American Samoa

The indigenous forest birds of American Samoa are increasingly threatened by changing patterns of rainfall and temperature that are associated with climate change as well as environmental stressors associated with agricultural and urban development, invasive species, and new introductions of avian diseases and disease vectors. Long term changes in their distribution, diversity, and population sizes could have significant impacts on the ecological integrity of the islands because of their critical role as pollinators and seed dispersers. We documented diversity of vector borne parasites on Tutuila and Ta‘u Islands over a 10-year period to expand earlier observations of Plasmodium, Trypanosoma , and filarial parasites, to provide better parasite identifications, and to create a better baseline for detecting new parasite introductions. We also identified potential mosquito vectors of avian Plasmodium and Trypanosoma , determined whether land clearing and habitat alterations associated with subsistence farming within the National Park of American Samoa can influence parasite prevalence, and determined whether parasite prevalence is correlated with seasonal changes in rainfall, temperature and wind speed. Three taxonomically distinct lineages of Plasmodium were identified from mosquito vectors and forest birds based on partial sequence data from parasite mitochondrial genes. All three have been described from passerine and galliform birds in Australasia. Two lineages, SCEDEN01 and ORW1, had elongate gametocytes and large schizonts that were consistent with species of Plasmodium in the subgenus Giavannolaia, but were taxonomically distinct from known morphological species of Plasmodium based on a Bayesian phylogenetic analysis of a 478 bp region of the parasite cytochrome b gene. Both are candidates for description as new species. The third lineage (GALLUS02) was detected only in mosquito vectors on Tutuila and was similar in cytochrome b sequence to P. juxtanucleare , a pathogenic species of Plasmodium from chickens and other galliform birds from Australasia, Africa, and South America. Plasmodium relictum , the malarial parasite that has had such a devastating impact on Hawaiian forest birds, was not detected. We observed large, striated trypanosomes in avian hosts from both Tutuila and Ta‘u Islands that fell within the same taxonomic clade as T. corvi and T. culicavium based on 18S ribosomal DNA sequence. We also observed sheathed microfilariae with pointed tails that had some morphological similarities to microfilaria from species of Pelecitus, Struthiofilaria and Eulimdana , but identification will require recovery and examination of adult filarial worms from the connective tissue or body cavities of infected birds. We also observed one or more species of haemococcidians (Isospora, synonym = Atoxoplasma ) within circulating lymphocytes from multiple avian host species. Overall prevalence of Plasmodium was higher on Ta‘u (22%, 75/341) than Tutuila (9.2%, 27/294), with most infections occurring in Polynesian starlings, Samoan starlings, Wattled honeyeaters, and Cardinal honeyeaters. Prevalence was relatively constant from year to year and between seasons at individual study sites, but varied among study sites, with highest rates of infection in areas with agricultural activity at Faleasao (37.4%, 73/195, Ta‘u Island) and Amalau Valley (9.7%, 21/216, Tutuila Island). Prevalence in more remote areas of the National Park of American Samoa was lower, ranging from 1.4% (2/146) at Laufuti and Luatele on Ta‘u to 7.7% (6/78) at Olo Ridge on Tutuila. Similar trends were evident for infections with Trypanosoma and filarial worms. Overall prevalence was not influenced significantly by warmer, wet (summer) or cooler, dry (winter) season. We detected Plasmodium infections in Culex sitiens and C. quinquefasciatus through either salivary gland and midgut dissections or PCR amplification of parasite cytochrome b genes in pooled or individual samples of mosquitoes that were collected on Tutuila. Pooled or individual Aedes oceanicus, A. polynesiensis, A. tutuilae, A. upolensis, A. nocturnus, Aedes (Finlaya) (mixed pools of A. samoanus, A. oceanicus, A. tutuilae), Aedes (Stegomyia) (mixed pools of A. aegypti, A. upolensis, A. polynesiensis ), and C. annulirostris were negative for Plasmodium , but we detected infections with Trypanosoma through midgut and salivary gland dissections in a single C. sitiens from Amalau Valley, Tutuila and three A. oceanicus from Faleasao, Ta‘u. Two of the A. oceanicus from Faleasao amplified successfully with Trypanosoma primers, but sequences were distinctly different from those obtained from avian hosts. We found a strong association between land use and prevalence of mosquito-transmitted parasites on Ta‘u Island with odds of being infected more than 20 times greater in agricultural plots than more remote native forest. This relationship was evident on Tutuila Island but not statistically significant because of the close proximity of study sites and observed movement of birds between native forest and agricultural land. Our data support previous studies that have suggested that Plasmodium and other vector-borne parasites are part of the indigenous parasite fauna in American Samoa. Transmission dynamics appear to be affected by environmental changes associated with land use practices.

HI