Geology topics

Tim L. King

Publications and source records attributed to Tim L. King.

At least 19 recordsLinked to original sources

USGS2022

Population genetics of Brook Trout (Salvelinus fontinalis) in the southern Appalachian Mountains

Broad-scale patterns of genetic diversity for Brook Trout remain poorly understood across their endemic range in the eastern United States. We characterized variation at 12 microsatellite loci in 22,020 Brook Trout among 836 populations from Georgia, USA to Quebec, Canada to the western Great Lakes region. Within-population diversity was typically lower in the southern Appalachians relative to the mid-Atlantic and northeastern regions. Effective population sizes in the southern Appalachians were often very small, with many estimates less than 30 individuals. The population genetics of Brook Trout in the southern Appalachians are far more complex than a conventionally held simple “northern” versus “southern” dichotomy would suggest. Contemporary population genetic variation was consistent with geographic expansion of Brook Trout from Mississippian, mid-Atlantic, and Acadian glacial refuges, as well as differentiation among drainages within these broader clades. Genetic variation was pronounced among drainages (57.4% of overall variation occurred among Hydrologic Unit Code (HUC)10 or larger units) but was considerable even at fine spatial scales (13% of variation occurred among collections within HUC12 drainage units). Remarkably, 87.2% of individuals were correctly assigned to their collection of origin. While comparisons with fish from existing major hatcheries showed impacts of stocking in some populations, genetic introgression did not overwhelm the signal of broad-scale patterns of population genetic structure. Although our results reveal deep genetic structure in Brook Trout over broad spatial extents, fine-scale population structuring is prevalent across the southern Appalachians. Our findings highlight the distinctiveness and vulnerability of many Brook Trout populations in the southern Appalachian Mountains and have important implications for wild Brook Trout management. To facilitate application of our findings by conservation practitioners, we provide an interactive online visualization tool to allow our results to be explored at management relevant scales.

North Carolina, Virginia, West Virginia↗

USGS2021

Genetic structure of Maryland Brook Trout populations: Management implications for a threatened species

Brook Trout Salvelinus fontinalis have declined across their native range due to multiple anthropogenic factors, including landscape alteration and climate change. Although coldwater streams in Maryland (eastern United States) historically supported significant Brook Trout populations, only fragmented remnant populations remain, with the exception of the upper Savage River watershed in western Maryland. Using microsatellite data from 38 collections, we defined genetic relationships of Brook Trout populations in Maryland drainages. Microsatellite analyses of Brook Trout indicated the presence of five major discrete units defined as the Youghiogheny (Ohio), Susquehanna, Patapsco/Gunpowder, Catoctin, and Upper Potomac, with a distinct genetic subunit present in the Savage River (upper Potomac). We did not observe evidence for widespread hatchery introgression with native Brook Trout. However, genetic effects due to fragmentation were evident in several Maryland Brook Trout populations, resulting in erosion of diversity that may have negative implications for their future persistence. Our current study supplements an increasing body of evidence that Brook Trout populations in Maryland are highly susceptible to multiple anthropogenic stresses, and many populations may be extirpated in the near future. Future management efforts focused on habitat protection and potential stream restoration, coupled with a comprehensive assessment framework that includes genetic considerations, may provide the best outlook for Brook Trout populations in Maryland.

Maryland↗

USGS2020

Assessment of Ambystomatid salamander populations and their breeding habitats in the Delaware Water Gap National Recreation Area

This report presents abundance and occurrence data for three species of ambystomad salamanders ( Ambystoma maculatum, A. jeffersonianum, and A. opacum ) collected over a 3-year period (2000, 2001, and 2002) at 200 potentional breeding sies within the Delaware Water Gap National Recreation Area (DEWA). In addition, numerous measures of inpond, near-pond, and landscape attributes were measured and used to inform statistical models to determine species-habitat relationships in the DEWA. The results of a 3-year study of ambystomatid salamander breeding habits and habitats in the (DEWA) that was conducted by the U.S. Geological Survey, in cooperation with the National Park Service, are described in the report. The objectives of the study were to document the population status and critical breeding habitats of the three species of ambystomatid salamanders known to be present in the DEWA— Ambystoma maculatum (spotted salamander), A. opacum (marbled salamander), and A. jeffersonianum (Jefferson salamander). DEWA managers are interested in ecological information on these species for several reasons. First, at the time the study began, there was little known regarding the status of pond-breeding amphibians and their habitats in the DEWA. Second, because they require undegraded habitats in both terrestrial and aquatic habitats to successfully complete their life cycles, the status of ambystomatid salamanders is widely viewed as indicative of overall ecosystem health. Third, because ambystomatid salamanders and other pond-breeding amphibians have been observed in numerous artificial impoundments with the DEWA, park managers would like to assess whether dismantling or discontinuing maintenance of artificial impoundments could affect pond-breeding amphibians and possibly other species that use pond or wetland habitats in the Park. In 2001, 2002, and 2003, the size and location of 200 wetlands, ponds, and artificial impoundments, and related landscape positions (Ridge versus Valley; Pennsylvania side versus New Jersey side of the Delaware river) were mapped, and site habitat data relating to salamander occurrence and abundance patterns were collected. The data collected during this study provide important new baseline information on ambystomatid salamanders and wetland habitats in the DEWA that will enhance long-term inventory and monitoring efforts. In addition, breeding habitat assessments indicate that ambystomatid salamanders may be sensitive to a wide variety of stresses important in the DEWA and in the region. In particular, recent trends in development (for example, roads) in and near the DEWA, regional increases in the acidity of precipitation, and predicted long-term warming trends for the region could be detrimental to pond-breeding salamander populations because of their effects on breeding site quality and quantity, and on the integrity of migration corridors. In contrast, the results of the study indicate management plans to eliminate small impoundments are not likely to adversely affect salamanders in DEWA, at least in the short-term. However, it is possible that these small impoundments may offer stable habitats that provide a rescure effect during long-term droughts.

New Jersey, Pennsylvania↗

USGS2020

Development of microsatellite markers for three at risk tiger beetles Cicindela dorsalis dorsalis, C. d. media, and C. puritana

Objective Tiger beetles inhabiting sandy beaches and cliffs along the east coast of the United States are facing increasing habitat loss due to erosion, urbanization, and sea level rise. The northeastern beach tiger beetle Cicindela dorsalis dorsalis and Puritan tiger beetle Cicindela puritana are both listed as threatened under the Endangered Species Act of 1973, while the white beach tiger beetle Cicindela dorsalis media is not listed but has been declining. Extirpation of these beetles, in some cases from entire states, has isolated many populations reducing gene flow and elevating the risk for the loss of genetic variation. To facilitate investigations of population genetic structure, we developed suites of microsatellite loci for conservation genetic studies. Results Shotgun genomic sequencing of all species identified thousands of candidate microsatellite loci, among which 17 loci were optimized and verified to cross-amplify within C. d. media and C. d. dorsalis , and eight separate loci were optimized for C. puritana . Most loci conformed to Hardy–Weinberg equilibrium, showed no evidence of linkage disequilibrium or null alleles, and revealed population genetic characteristics informative for natural resource managers among the populations tested.

BMC Research Notes↗

USGS2020

Using advanced population genomics to better understand the relationship between offshore and spawning habitat use for Atlantic Sturgeon

Atlantic Sturgeon ( Acipenser oxyrinchus oxyrinchus ) are a large-bodied anadromous fish that historically supported important fisheries along the east coast of the United States. Following years of overharvest and habitat degradation, populations experienced severe declines. In 2012, the National Marine Fisheries Service listed Atlantic Sturgeon under the Endangered Species Act (ESA; 61 FR 4722). Their listing named five Distinct Population Segments (DPSs), predicated on genetic groups composed of geographically proximate populations. Federal management of Atlantic Sturgeon presents challenges, as sturgeon from each of the five DPSs mix extensively in coastal and marine habitats yet take and recovery progress must be evaluated separately for each unit. Genetic assignment testing based on mitochondrial and microsatellite markers allows individuals to be assigned back to their natal river and DPS. However, this approach is not perfect and some individuals may be incorrectly assigned. Recent advances in genomics offer the potential of a higher resolution approach to genetic assignment testing, and thus may reduce uncertainty associated with assignment testing. In addition, genomics allows a greater number of markers to be examined from across a broader portion of the sturgeon genome, thus may provide an enhanced perspective of population structure for the species, and potentially allow other previously intractable questions to be addressed (Bernatchez et al. 2017, Supple and Shapiro 2018). We used next-generation sequencing to develop a draft genome for Atlantic Sturgeon and identify single nucleotide polymorphisms (SNPs) that could be used to resolve the natal river and DPS of individual Atlantic Sturgeon. We identified 1,210 candidate SNPs within the nuclear genome as well as 49 SNPs within the mitochondrial genome. After filtering and review, we selected 161 nuclear SNPs and 39 mitochondrial SNPs for further testing and evaluation. We used genotyping-in-thousands by sequencing (GT-seq) to simultaneously sequence nuclear SNP loci, mitochondrial SNP loci, and the existing panel of twelve microsatellite loci. This effort required a pilot sequencing run on a single sturgeon sample to test marker amplification and refine primer strengths, followed by a series of sequencing runs to generate baseline data for 288 individuals representing nine populations of Atlantic Sturgeon in four DPSs. Using baseline data from the nine populations, we ran a series of genomic analyses to characterize diversity within and among populations, providing a benchmark for this species using the new SNP markers. Allelic richness was similar for all populations, although there was a general trend of more northern population containing greater levels of allelic richness. Interestingly, we observed linkage disequilibrium among many pairs of loci within many populations. This might be the result of physical linkage but could also suggest these populations are recovering from genetic bottlenecks and/or are effectively small, leading to specific haplotypes to be favored by chance. Pairwise differentiation among populations varied among the populations ( F ST range: 0.010-0.098) and was significantly correlated ( r = 0.771; P < 0.001) to pairwise F ST observed using microsatellite markers). Population clustering and ordination techniques using the new genomic data both support an overall population structure that is similar to the current DPS management units (which were developed primarily based on microsatellite genetic data). Overall, this suggests that existing microsatellite markers and the panel of SNP markers developed in this study provide similar information about the populations structure and ecology of Atlantic Sturgeon. Given the observed differences in allele frequencies among populations, our genomic baseline supports previous assertations that Atlantic Sturgeon show natal homing, despite mixing extensively in marine waters during non-breeding periods. Lower levels of differentiation between populations in the South Atlantic DPS suggest that populations in this region may have greater levels of gene flow relative to their more northerly conspecifics, which has also previously been suggested based on microsatellite data. The observed differentiation among populations provides the necessary foundation for determining the natal river and DPS of Atlantic Sturgeon using assignment testing. We tested the utility of our new genomic baseline for resolving the population and DPS of Atlantic Sturgeon. Our nuclear SNP markers showed utility for identifying the origin of unknown Atlantic Sturgeon samples, as 86.5% were assigned to the correct DPS and 66.3% were assigned to the correct natal river. However, since this study was funded the Conservation Genetics and Genomics Laboratory at Leetown Science Center has made significant improvements to their microsatellite genetic baseline, which now performs more effectively than our new genomic approach (the genetic baseline includes 12 populations and 5 DPSs, and correctly assigns 95.8% of individuals to DPS and 84.9% of individuals to their natal population using 12 microsatellite loci). We conducted an ad hoc exploration of how additional microsatellite or nuclear SNP loci may further improve the accuracy of assignment testing. We found that additional microsatellite markers are likely to result in greater improvements in assignment efficiency than additional nuclear SNPs. However, a much larger number of SNP loci (which if identified could be sequenced using other methods that are now available; e.g., the RAD-capture approach published by Ali et al. 2016) could produce assignment efficiencies that are greater than what is currently feasible using microsatellites. In the absence of further research and development of additional SNP markers for Atlantic Sturgeon (possibly using an approach other than GT-seq), the existing microsatellite loci are the most effective means available to determine the natal river and DPS of Atlantic Sturgeon encountered in offshore waters. Because our new genomic markers were less effective than the existing panel of 12 microsatellite markers, we chose to use the existing microsatellite markers to assign Atlantic Sturgeon captured in another BOEM-funded study (cooperative agreement M16AC00003; Monitoring endangered Atlantic Sturgeon and commercial finfish habitat use offshore New York) following consultation with our project officer. Using this approach, we genotyped and assigned 186 Atlantic Sturgeon captured in coastal waters off the Rockaway Peninsula, New York. The vast majority of these sturgeon were assigned to the New York Bight DPS (94.62%), and most appear to belong to the Hudson River population (87.10%) with smaller contributions from the Delaware River population (7.53%). Smaller contributions (2.15%) were observed from six other populations, including those from the James, York, Kennebec, Ogeechee, and Edisto rivers. Although most of the fish we assigned were assigned to the nearest spawning rivers (Hudson and Delaware), the contributions from distant rivers is consistent with the propensity of this species to move long distances and form mixed stock aggregations along the continental shelf. This finding indicates that spawning populations (and their corresponding DPS) from distant locations may potentially be impacted by offshore activities. In fact, activities in this region of the New York Bight could negatively impact Atlantic Sturgeon population from at least four different DPSs. Genetic or genomic assignment testing remains an essential tool to characterize potential impacts to Atlantic Sturgeon populations and should be applied more broadly to better characterize potential impacts of activities in other locations.

Atlantic Coast↗

USGS2019

Understanding the genetic characteristics of Wild Brook Trout populations in North Carolina thanks to the guidance of Dr. Tim King

We genotyped 7,588 brook trout representing 406 collections from across the State of North Carolina (Figure 1) at 12 microsatellite loci (King et al. 2012). The vast majority of collections appeared to represent single populations, based on general conformance to HardyWeinberg equilibrium and limited evidence for linkage-disequilibrium. Allelic diversity was low to moderate relative to Brook Trout Salvelinus fontinalis populations endemic to higher latitudes. Effective population sizes varied widely among populations, but were often very small and indicate that many populations are at risk of losing diversity through genetic drift. Remarkable levels of genetic differentiation exist among populations, which suggests that little, if any, gene flow occurs among most populations. Analysis of molecular variance (AMOVA) revealed that a substantial portion of the observed genetic variation was attributed to differences among patches (44.8%), and there was some variation (11.2%) even among collections within a single patch. These results, taken in conjunction with high levels of genetic differentiation among populations, suggest that the fundamental unit of management for Brook Trout should be the population. Interestingly, despite extensive stocking across the state, the vast majority of wild populations show limited evidence of introgression by northern origin hatchery strains. These results represent a valuable baseline for management and restoration efforts, and can be used to (a) select suitable donor streams for translocation efforts, (b) identify streams with low effective population sizes that may be vulnerable to extirpation, and (c) target stocking efforts into watersheds where extensive introgression has already occurred. All data associated with this manuscript has been publicly released (Kazyak et al. 2017).

North Carolina↗

USGS2018

Neutral genetic and phenotypic variation within and among isolated headwater Brook Trout populations

Isolated populations are challenging to manage and conserve as they are particularly vulnerable to genetic drift, allelic fixation, inbreeding, and may express markedly reduced phenotypic variability. We sought to improve our understanding of how spatial isolation, occupancy range, and restricted gene flow influence contemporary phenotypic variation within and among native populations of Brook Trout Salvelinus fontinalis by examining the neutral genetic and phenotypic characteristics of 35 isolated headwater populations from Great Smoky Mountains National Park. Across a suite of 13 neutral microsatellite loci, we observed high levels of allelic fixation and considerable genetic differentiation among populations, subwatersheds, and watersheds that were consistent with patterns of isolation. We observed significant, positive correlations between allelic diversity and estimates of effective population sizes. In contrast, we observed considerably less phenotypic structure among streams, subwatersheds, and watersheds. Much of the phenotypic variation observed occurred among individuals within populations. Pairwise Mann‐Whitney tests revealed no significant phenotypic differences among the populations of Brook Trout we examined. Similarly, we observed no significant relationship between the amount of phenotypic variation within populations and any of the examined measures of genetic diversity or the amount of occupied habitat sampled, which suggests that unmeasured variables may be influencing morphometric and meristic variation within isolated populations. The observed patterns of isolation, genetic drift, and allelic fixation highlight the importance of enhancing population connectivity, but also suggest considerable phenotypic variability may persist within small, fragmented populations. Our results elucidate some challenges associated with managing and conserving isolated populations of Brook Trout, and reinforce the importance of conducting genetic studies on fragmented populations to inform management decisions.

Transactions of the American Fisheries Society↗

USGS2018

The complete maternal mitochondrial genome sequences of two imperiled North American freshwater mussels: Alasmidonta heterodon and Alasmidonta varicosa (Bivalvia: Unionoida: Unionidae)

The freshwater mussels Alasmidonta heterodon and A. varicosa historically inhabited rivers along the North American Atlantic coast from the Carolinas, U.S.A., to New Brunswick, CA. However, many populations have been extirpated, and A. heterodon is now federally listed in the U.S.A. as endangered, and both A. heterodon and A. varicosa are listed as vulnerable on the IUCN Red List. To facilitate genetic study of these species, we sequenced the complete female mitochondrial genomes of A. heterodon (15,909 bp; GenBank accession no. MG905826), and A. varicosa (15,693 bp; GenBank accession no. MG938673). Both mitogenomes contained 14 protein coding genes, 2 rRNA genes, and 22 tRNAs with the same gene order as reported for other members of the subfamily Anodontinae. When these two genomes were put into a phylogenetic context with other members of the Unionidae, they clustered together with other species in the subfamily Anodontinae, Tribe Anodontini.

Mitochondrial DNA Part B↗

USGS2018

The complete mitochondrial genome of the stalk-forming diatom Didymosphenia geminata

The complete mitogenome of the stalk-forming diatom Didymosphenia geminata collected from Mineral County, WV, USA was sequenced on the Ion Torrent PGM and Proton sequencers. The D. geminata mitogenome is 37,765 bp and encodes 35 protein coding genes, 25 tRNAs, and both large and small subunit ribosomal RNA genes. The nad 11 gene is split into two domains as observed in Phaeodactylum tricornutum , and D. geminata also lacks the large repeat region found in the P. tricornutum mitogenome. Gene order and content within the D. geminata mitogenome is similar to the diatom Berkeleya fennica .

Mitochondrial DNA Part B↗

USGS2017

Assessing the impact of stocking northern-origin hatchery brook trout on the genetics of wild populations in North Carolina

The release of hatchery-origin fish into streams with endemics can degrade the genetics of wild populations if interbreeding occurs. Starting in the 1800s, brook trout descendent from wild populations in the northeastern United States were stocked from hatcheries into streams across broad areas of North America to create and enhance fishery resources. Across the southeastern United States, many millions of hatchery-origin brook trout have been released into hundreds of streams, but the extent of introgression with native populations is not well resolved despite large phylogeographic distances between these groups. We used three assessment approaches based on 12 microsatellite loci to examine the extent of hatchery introgression in 406 wild brook trout populations in North Carolina. We found high levels of differentiation among most collections (mean F ′ ST = 0.718), and among most wild collections and hatchery strains (mean F ′ ST = 0.732). Our assessment of hatchery introgression was consistent across the three metrics, and indicated that most wild populations have not been strongly influenced by supplemental stocking. However, a small proportion of wild populations in North Carolina appear to have been strongly influenced by stocked conspecifics, or in some cases, may have been founded entirely by hatchery lineages. In addition, we found significant differences in the apparent extent of hatchery introgression among major watersheds, with the Savannah River being the most strongly impacted. Conversely, populations in the Pee Dee River watershed showed little to no evidence of hatchery introgression. Our study represents the first large-scale effort to quantify the extent of hatchery introgression across brook trout populations in the southern Appalachians using highly polymorphic microsatellite markers.

North Carolina↗

USGS2017

Microsatellite marker development from next-generation sequencing in the New England cottontail (Sylvilagus transitionalis) and cross-amplification in the eastern cottontail (S. floridanus)

Objective The New England cottontail ( Sylvilagus transitionalis ) is a species of high conservation priority in the Northeastern United States, and was a candidate for federal listing under the Endangered Species Act until a recent decision determined that conservation actions were sufficient to preclude listing. The aim of this study was to develop a suite of microsatellite loci to guide future research efforts such as the analysis of population genetic structure, genetic variation, dispersal, and genetic mark-recapture population estimation. Results Thirty-five microsatellite markers containing tri- and tetranucleotide sequences were developed from shotgun genomic sequencing of tissue from S. transitionalis , S. obscurus , and S. floridanus . These loci were screened in n = 33 wild S. transitionalis sampled from a population in eastern Massachusetts, USA. Thirty-two of the 35 loci were polymorphic with 2–6 alleles, and observed heterozygosities of 0.06–0.82. All loci conformed to Hardy–Weinberg Equilibrium proportions and there was no evidence of linkage disequilibrium or null alleles. Primers for 33 of the 35 loci amplified DNA extracted from n = 6 eastern cottontail ( S. floridanus ) samples, of which nine revealed putative species-diagnostic alleles. These loci will provide a useful tool for conservation genetics investigations of S. transitionalis and a potential diagnostic species assay for differentiating sympatric eastern and New England cottontails.

Massachusetts↗

USGS2017

Life history and status of Shortnose Sturgeon (Acipenser brevirostrum LeSueur, 1818)

Shortnose Sturgeon = SNS (Acipenser brevirostrum) is a small diadromous species with most populations living in large Atlantic coast rivers and estuaries of North America from New Brunswick, Canada, to GA, USA. There are no naturally landlocked populations, so all populations require access to fresh water and salt water to complete a natural life cycle. The species is amphidromous with use of fresh water and salt water (the estuary) varied across the species range, a pattern that may reflect whether freshwater or saltwater habitats provide optimal foraging and growth conditions. Migration is a dominant behavior during life history, beginning when fish are hatchling free embryos (southern SNS) or larvae (northeastern and far northern SNS). Migration continues by juveniles and nonspawning adult life stages on an individual time schedule with fish moving between natal river and estuary to forage or seek refuge, and by spawning adults migrating to and from riverine spawning grounds. Coastal movements by adults throughout the range (but particularly in the Gulf of Maine = GOM and among southern rivers) suggest widespread foraging, refuge use, and widespread colonization of new rivers. Colonization may also be occurring in the Potomac River, MD–VA–DC (midAtlantic region). Genetic studies (mtDNA and nDNA) identified distinct individual river populations of SNS, and recent rangewide nDNA studies identified five distinct evolutionary lineages of SNS in the USA: a northern metapopulation in GOM rivers; the Connecticut River; the Hudson River; a Delaware River–Chesapeake Bay metapopulation; and a large southern metapopulation (SC rivers to Altamaha River, GA). The Saint John River, NB, Canada, in the Bay of Fundy (north of the GOM), is the sixth distinct genetic lineage within SNS. Life history information from telemetry tracking supports the genetic information documenting extensive movement of adults among rivers within the three metapopulations. However, individual river populations with spawning adults are still the best basal unit for management and recovery planning. The focus on individual river populations should be complemented with attention to migratory processes and corridors that foster metapopulation level risks and benefits. The species may be extirpated at the center of the range, i.e., the midAtlantic region (Chesapeake Bay, MD–VA, and probably, NC), but large rivers in VA, including the James and Potomac rivers, need study. The largest SNS populations in GOM and northeastern rivers, like the Kennebec, Hudson, and Delaware rivers, typically have tens of thousands of adults. This contrasts with southern rivers, where rivers typically have much fewer (<2500) adults, except for the Altamaha River (>6000 adults). River damming in the 19th and 20th Centuries extirpated some populations, and also, created two dysfunctional segmented populations: the Connecticut River SNS in CT–MA and the SanteeCooper rivers–Lake Marion SNS in SC. The major anthropogenic impact on SNS in marine waters is fisheries bycatch. The major impacts that determine annual recruitment success occur in freshwater firstly, where adult spawning migrations and spawning are blocked or spawning success is affected by river regulation and secondly, where poor survival of early life stages is caused by river dredging, pollution, and unregulated impingement/entrainment in water withdrawal facilities. Climate warming has the potential to reduce abundance or eliminate SNS in many rivers, particularly in the South. In 1998, the National Marine Fisheries Service (NMFS) recommended management of 19 rivers as distinct population segments (DPSs) based on strong fidelity to natal rivers. A Biological Assessment completed in 2010 reaffirmed this approach. NMFS has not formally listed DPSs under the ESA and the species remains listed as endangered rangewide in the USA.

Journal of Applied Ichthyology↗

USGS2017

Between hot rocks and dry places: The status of the Dixie Valley toad

In Dixie Valley, Nevada, an isolated population of toads has been the subject of proactive conservation measures by the Nevada Department of Wildlife and the U.S. Fish and Wildlife Service since 2008 due to concerns about potential habitat degradation resulting from exploitation of nearby geothermal energy resources. These toads appear to belong within the Anaxyrus boreas species group but are commonly referred to as Dixie Valley toads (DVTs). The DVT is currently confined to an extremely narrow habitat range (370 ha) that is geographically isolated from any other A. boreas population. In this study, genetic variations in mitochondrial genes and 11 microsatellite loci were used to assess the affinities of DVTs in relation to members of the A. boreas species group. We compared results from DVTs with previously published data spanning much of the range of A. boreas in the United States and new data from a nearby toad population within Dixie Valley. Data from both mitochondrial DNA and microsatellites placed DVTs inside the A. boreas species group. In particular, DVTs fell into a cluster of A. boreas from Washington and California, along with other species from the A. boreas species group, namely A. nelsoni, A. canorus, and A. exsul. Genetic differentiation of DVTs was lowest between A. boreas populations in Washington and California. However, allele frequencies were significantly different between DVTs and all other populations, including a nearby locality within Dixie Valley. This genetic differentiation, along with the DVT's geographical isolation and restricted habitat, warrants recognition of the DVT as a distinct management unit.

Nevada↗

USGS2016

Conservation status of the American horseshoe crab, (Limulus polyphemus): A regional assessment

Horseshoe crabs have persisted for more than 200 million years, and fossil forms date to 450 million years ago. The American horseshoe crab ( Limulus polyphemus ), one of four extant horseshoe crab species, is found along the Atlantic coastline of North America ranging from Alabama to Maine, USA with another distinct population on the coasts of Campeche, Yucatán and Quintana Roo in the Yucatán Peninsula, México. Although the American horseshoe crab tolerates broad environmental conditions, exploitation and habitat loss threaten the species. We assessed the conservation status of the American horseshoe crab by comprehensively reviewing available scientific information on its range, life history, genetic structure, population trends and analyses, major threats, and conservation. We structured the status assessment by six genetically-informed regions and accounted for sub-regional differences in environmental conditions, threats, and management. The transnational regions are Gulf of Maine (USA), Mid-Atlantic (USA), Southeast (USA), Florida Atlantic (USA), Northeast Gulf of México (USA), and Yucatán Peninsula (México). Our conclusion is that the American horseshoe crab species is vulnerable to local extirpation and that the degree and extent of risk vary among and within the regions. The risk is elevated in the Gulf of Maine region due to limited and fragmented habitat. The populations of horseshoe crabs in the Mid-Atlantic region are stable in the Delaware Bay area, and regulatory controls are in place, but the risk is elevated in the New England area as evidenced by continuing declines understood to be caused by over-harvest. The populations of horseshoe crabs in the Southeast region are stable or increasing. The populations of horseshoe crabs in the Florida Atlantic region show mixed trends among areas, and continuing population reductions at the embayment level have poorly understood causes. Within the Northeast Gulf of Mexico, causes of population trends are poorly understood and currently there is no active management of horseshoe crabs. Horseshoe crabs within México have conservation protection based on limited and fragmented habitat and geographic isolation from other regions, but elevated risk applies to the horseshoe crabs in the Yucatán Peninsula region until sufficient data can confirm population stability. Future species status throughout its range will depend on the effectiveness of conservation to mitigate habitat loss and manage for sustainable harvest among and within regions.

Reviews in Fish Biology and Fisheries↗

USGS2016

Comparative mitogenomic analyses of three North American stygobiont amphipods of the genus Stygobromus (Crustacea: Amphipoda)

The mitochondrial genomes of three North American stygobiont amphipods Stygobromus tenuis potomacus , S. foliatus and S. indentatus collected from Caroline County, VA, were sequenced using a shotgun sequencing approach on an Illumina NextSeq500 (Illumina Inc., San Diego, CA). All three mitogenomes displayed 13 protein-coding genes, 22 tRNAs and two rRNAs typical of metazoans. While S. tenuis and S. indentatus displayed identical gene orders similar to the pancrustacean ground pattern, S. foliatus displayed a transposition of the trnL2 - cox2 genes to after atp8-atp6 . In addition, a short atp8 gene, longer rrnL gene and large inverted repeat within the Control Region distinguished S. foliatus from S. tenuis potomacus and S. indentatus . Overall, it appears that gene order varies considerably among amphipods, and the addition of these Stygobromus mitogenomes to the existing sequenced amphipod mitogenomes will prove useful for characterizing evolutionary relationships among various amphipod taxa, as well as investigations of the evolutionary dynamics of the mitogenome in general.

Mitochondrial DNA Part B↗

USGS2015

Metagenomic analysis of planktonic microbial consortia from a non-tidal urban-impacted segment of James River

Knowledge of the diversity and ecological function of the microbial consortia of James River in Virginia, USA, is essential to developing a more complete understanding of the ecology of this model river system. Metagenomic analysis of James River's planktonic microbial community was performed for the first time using an unamplified genomic library and a 16S rDNA amplicon library prepared and sequenced by Ion PGM and MiSeq, respectively. From the 0.46-Gb WGS library (GenBank:SRR1146621; MG-RAST:4532156.3), 4 × 10 6 reads revealed >3 × 10 6 genes, 240 families of prokaryotes, and 155 families of eukaryotes. From the 0.68-Gb 16S library (GenBank:SRR2124995; MG-RAST:4631271.3; EMB:2184), 4 × 10 6 reads revealed 259 families of eubacteria. Results of the WGS and 16S analyses were highly consistent and indicated that more than half of the bacterial sequences were Proteobacteria , predominantly Comamonadaceae . The most numerous genera in this group were Acidovorax (including iron oxidizers, nitrotolulene degraders, and plant pathogens), which accounted for 10 % of assigned bacterial reads. Polaromonas were another 6 % of all bacterial reads, with many assignments to groups capable of degrading polycyclic aromatic hydrocarbons. Albidiferax (iron reducers) and Variovorax (biodegraders of a variety of natural biogenic compounds as well as anthropogenic contaminants such as polycyclic aromatic hydrocarbons and endocrine disruptors) each accounted for an additional 3 % of bacterial reads. Comparison of these data to other publically-available aquatic metagenomes revealed that this stretch of James River is highly similar to the upper Mississippi River, and that these river systems are more similar to aquaculture and sludge ecosystems than they are to lakes or to a pristine section of the upper Amazon River. Taken together, these analyses exposed previously unknown aspects of microbial biodiversity, documented the ecological responses of microbes to urban effects, and revealed the noteworthy presence of 22 human-pathogenic bacterial genera (e.g., Enterobacteriaceae , pathogenic Pseudomonadaceae , and ‘ Vibrionales' ) and 6 pathogenic eukaryotic genera (e.g., Trypanosomatidae and Vahlkampfiidae). This information about pathogen diversity may be used to promote human epidemiological studies, enhance existing water quality monitoring efforts, and increase awareness of the possible health risks associated with recreational use of James River.

Virginia↗

USGS2015

Spatial structure of morphological and neutral genetic variation in Brook Trout

Brook Trout Salvelinus fontinalis exhibit exceptional levels of life history variation, remarkable genetic variability, and fine-scale population structure. In many cases, neighboring populations may be highly differentiated from one another to an extent that is comparable with species-level distinctions in other taxa. Although genetic samples have been collected from hundreds of populations and tens of thousands of individuals, little is known about whether differentiation at neutral markers reflects phenotypic differences among Brook Trout populations. We compared differentiation in morphology and neutral molecular markers among populations from four geographically proximate locations (all within 24 km) to examine how genetic diversity covaries with morphology. We found significant differences among and/or within streams for all three morphological axes examined and identified the source stream of many individuals based on morphology (52.3% classification efficiency). Although molecular and morphological differentiation among streams ranged considerably (mean pairwise F ST : 0.023–0.264; pairwise P ST : 0.000–0.339), the two measures were not significantly correlated. While in some cases morphological characters appear to have diverged to a greater extent than expected by neutral genetic drift, many traits were conserved to a greater extent than were neutral genetic markers. Thus, while Brook Trout exhibit fine-scale spatial patterns in both morphology and neutral genetic diversity, these types of biological variabilities are being structured by different ecological and evolutionary processes. The relative influences of genetic drift versus selection and phenotypic plasticity in shaping morphology appear to vary among populations occupying nearby streams.

Maryland↗

USGS2015

Isolation and characterization of microsatellite DNA loci in the threatened flat-spired three-toothed land snail Triodopsis platysayoides

The hermaphroditic flat-spired three-tooth land snail ( Triodopsis platysayoides ) is endemic to a 21-km stretch of the Cheat River Gorge of northeastern West Virginia, USA. We document isolation and characterization of ten microsatellite DNA markers in this at-risk species. The markers displayed a moderate level of allelic diversity (averaging 7.1 alleles/locus) and heterozygosity (averaging 58.6 %). Allelic diversity at seven loci was sufficient to produce unique multilocus genotypes; no indication of selfing was detected in this cosexual species. Minimal deviations from Hardy–Weinberg equilibrium and no linkage disequilibrium were observed within subpopulations. All loci deviated from Hardy–Weinberg expectations when individuals from subpopulations were pooled. Microsatellite markers developed for T . platysayoides yielded sufficient genetic diversity to (1) distinguish all individuals sampled and the level of selfing; (2) be appropriate for addressing fine-scale population structuring; (3) provide novel demographic insights for the species; and (4) cross-amplify and detect allelic diversity in the congeneric T. juxtidens .

Conservation Genetics Resources↗