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T.L. King

Publications and source records attributed to T.L. King.

At least 19 recordsLinked to original sources
USGS2013

Differential expression profiles of microRNA in the little brown bat (Myotis lucifugus) associated with white nose syndrome affected and unaffected individuals

First documented in New York State in 2006, white nose syndrome (WNS) quickly became the leading cause of mortality in hibernating bat species in the United States. WNS is caused by a psychrophilic fungus, Geomyces destructans. Clinical signs of this pathogen are expressed as a dusty white fungus predominately around the nose and on the wings of affected bats. Relatively new biomarkers, such as microRNAs (miRNAs) are being targeted as markers to predict the syndrome prior to the clinical manifestation. The primary objective of this study was to identify miRNAs that could serve as biomarkers and proxies of little brown bat health. Bats were collected from hibernacula that had tested positive and negative for WNS. Genetic sequencing was completed using the Ion Torrent platform. A number of miRNAs were identified from the liver as putative biomarkers of WNS. However, given the small sample size for each treatment, this data set has only coarsely identified miRNAs indicative of WNS, and further validation is required.

Open-File Report↗
USGS2011

Demographic and genetic status of an isolated population of bog turtles (Glyptemys muhlenbergii): Implications for managing small populations of long-lived animals

In this study, we sought to determine the population stability and genetic diversity of one isolated population of the federally-threatened bog turtle ( Glyptemys muhlenbergii ) in North Carolina. Using capture–recapture data, we estimated adult survival and population growth rate from 1992 to 2007. We found that the population decreased from an estimated 36 adult turtles in 1994 to approximately 11 adult turtles in 2007. We found a constant adult survival of 0.893 (SE = 0.018, 95% confidence interval, 0.853–0.924) between 1992 and 2007. Using 18 microsatellite markers, we compared the genetic status of this population with five other bog turtle populations. The target population displayed allelic richness (4.8 ± 0.5) and observed heterozygosity (0.619 ± 0.064) within the range of the other bog turtle populations. Coalescent analysis of population growth rate, effective population size, and timing of population structuring event also indicated the genetics of the target population were comparable to the other populations studied. Estimates of effective population size were a proportion of the census size in all populations except the target population, in which the effective population size was larger than the census size (30 turtles vs. 11 turtles). We attribute the high genetic diversity in the target population to the presence of multiple generations of old turtles. This study illustrates that the demographic status of populations of long-lived species may not be reflected genetically if a decline occurred recently. Consequently, the genetic integrity of populations of long-lived animals experiencing rapid demographic bottlenecks may be preserved through conservation efforts effective in addressing demographic problems.

Conservation Genetics↗
USGS2011

Tools for assessing kinship, population structure, phylogeography, and interspecific hybridization in Asian carps invasive to the Mississippi River, USA: isolation and characterization of novel tetranucleotide microsatellite DNA loci in silver carp Hypophthalmichthys molitrix

We document the isolation and characterization of novel tetranucleotide microsatellite DNA markers for the invasive silver carp Hypophthalmichthys molitrix and provide the results of cross-species amplification for three additional invasive carp species: bighead (H. nobilis), grass (Ctenopharyngodon idella) and black (Mylopharyngodon piceus). In the target species these markers yielded levels of allelic diversity (average 4.4 alleles/locus) and heterozygosity (average 54.7%) sufficient to: (1) provide unique multilocus genotypes; (2) delineate kinship relationships; (3) differentiate populations/species; (4) estimate effective population sizes; and (5) provide unique demographic perspectives for control or eradication. Currently these markers are being utilized to determine the degree of introgressive hybridization between H. molitrix and H. nobilis, to quantify gene flow between different sub-basins established in the central United States, and to assess the demographic status of sub-basin groups. This information will be critically important in the management/control of these invasive species.

Mississippi River↗
USGS2010

Challenges of DNA-based mark-recapture studies of American black bears

We explored whether genetic sampling would be feasible to provide a region-wide population estimate for American black bears ( Ursus americanus ) in the southern Appalachians, USA. Specifically, we determined whether adequate capture probabilities ( p >0.20) and population estimates with a low coefficient of variation (CV <20%) could be achieved given typical agency budget and personnel constraints. We extracted DNA from hair collected from baited barbed-wire enclosures sampled over a 10-week period on 2 study areas: a high-density black bear population in a portion of Great Smoky Mountains National Park and a lower density population on National Forest lands in North Carolina, South Carolina, and Georgia. We identified individual bears by their unique genotypes obtained from 9 microsatellite loci. We sampled 129 and 60 different bears in the National Park and National Forest study areas, respectively, and applied closed mark&ndash;recapture models to estimate population abundance. Capture probabilities and precision of the population estimates were acceptable only for sampling scenarios for which we pooled weekly sampling periods. We detected capture heterogeneity biases, probably because of inadequate spatial coverage by the hair-trapping grid. The logistical challenges of establishing and checking a sufficiently high density of hair traps make DNA-based estimates of black bears impractical for the southern Appalachian region. Alternatives are to estimate population size for smaller areas, estimate population growth rates or survival using mark&ndash;recapture methods, or use independent marking and recapturing techniques to reduce capture heterogeneity.

Georgia, North Carolina, South Carolina, Tennessee↗
USGS2008

Characterization of 13 microsatellite loci for the deep-sea coral, Lophelia pertusa (Linnaeus 1758), from the western North Atlantic Ocean and Gulf of Mexico

A suite of 13 polymorphic tri- and tetranucleotide microsatellite loci were isolated from the ahermatypic deep-sea coral, Lophelia pertusa . Among 51 individuals collected from three disjunct oceanic regions, allelic diversity ranged from six to 38 alleles and averaged 9.1 alleles per locus. Observed heterozygosity ranged from 9.1 to 96.8% and averaged 62.3% in the Gulf of Mexico population. For some loci, amplification success varied among collections, suggesting regional variation in priming site sequences. Four loci showed departures from Hardy–Weinberg equilibrium in certain collections which may reflect nonrandom mating.

Molecular Ecology Resources↗
USGS2008

Microsatellite DNA primers for the candy darter, Etheostoma osburni and variegate darter, Etheostoma variatum, and cross-species amplification in other darters (Percidae)

In order to investigate a potential hybrid zone between the candy darter, Etheostoma osburni, and variegate darter, Etheostoma variatum, and examine population variation within E. osburni, a suite of primers for 15 polymorphic microsatellite loci were developed. The average number of alleles per locus was 5.5 in E. osburni and 7.6 in E. variatum, and the average observed heterozygosities were 62.5% and 71.4%, respectively. There were no deviations from Hardy-Weinberg equilibrium and no observed linkage disequilibrium after Bonferroni correction. The utility of these primers was also tested in 11 species of darters representing all four genera of darters. Success of cross-species amplification was largely consistent with phylogenetic relationships of darters. ?? 2007 The Authors.

Molecular Ecology Resources↗
USGS2008

Tracing the first steps of American sturgeon pioneers in Europe

Background. A Baltic population of Atlantic sturgeon was founded ???1,200 years ago by migrants from North America, but after centuries of persistence, the population was extirpated in the 1960s, mainly as a result of over-harvest and habitat alterations. As there are four genetically distinct groups of Atlantic sturgeon inhabiting North American rivers today, we investigated the genetic provenance of the historic Baltic population by ancient DNA analyses using mitochondrial and nuclear markers. Results. The phylogeographic signal obtained from multilocus microsatellite DNA genotypes and mitochondrial DNA control region haplotypes, when compared to existing baseline datasets from extant populations, allowed for the identification of the region-of-origin of the North American Atlantic sturgeon founders. Moreover, statistical and simulation analyses of the multilocus genotypes allowed for the calculation of the effective number of individuals that originally founded the European population of Atlantic sturgeon. Our findings suggest that the Baltic population of A. oxyrinchus descended from a relatively small number of founders originating from the northern extent of the species' range in North America. Conclusion. These results demonstrate that the most northerly distributed North American A. oxyrinchus colonized the Baltic Sea ???1,200 years ago, suggesting that Canadian specimens should be the primary source of broodstock used for restoration in Baltic rivers. This study illustrates the great potential of patterns obtained from ancient DNA to identify population-of-origin to investigate historic genotype structure of extinct populations. ?? 2008 Ludwig et al; licensee BioMed Central Ltd.

BMC Evolutionary Biology↗
USGS2008

Conservation genetics and species recovery

Recent advances in molecular genetics have proven to be extremely useful in efforts to conserve imperiled species. Genetics data are used to identify appropriate units of management (e.g., populations, metapopulations), effective sizes of breeding populations, population mixing rates, and other variables. These data help managers make decisions about which populations to preserve, whether to move individuals from one site to another, how to breed species most effectively in captivity, and even, in some cases, what taxonomic classification is most appropriate. Many U.S. Geological Survey (USGS) Science Centers and Cooperative Fish and Wildlife Research Units have developed capabilities in genetics research. The two case studies that follow illustrate how USGS geneticists are assisting managers in recovering species on the brink.

Endangered Species Bulletin↗
USGS2006

Isolation and characterization of microsatellite loci in Alasmidonta heterodon (Bivalvia: Unionidae)

We developed 13 species-specific microsatellite markers for the federally endangered Atlantic slope unionid Alasmidonta heterodon. Four to 18 alleles per locus were observed among 30 individuals. Observed heterozygosity throughout the loci ranged from 26.9 to 86.2% and averaged 63.6%. Estimates of individual pairwise genetic distances indicated that levels of genetic diversity among loci were sufficient to produce unique multilocus genotypes for all animals surveyed. Randomization tests showed that genotypes for this collection were consistent with Hardy-Weinberg expectations, and no significant linkage disequilibrium was observed between loci. These loci therefore appear suitable for population surveys, kinship assessment and other such applications. ?? 2006 Blackwell Publishing Ltd.

Molecular Ecology Notes↗
USGS2006

Phylogeographic analyses suggest multiple lineages of Crystallaria asprella (Percidae: Etheostominae)

The crystal darter, Crystallaria asprella , exists in geographically isolated populations that may be glacial relicts from its former, wide distribution in the Eastern U.S. An initial phylogeographic survey of C. asprella based upon the mitochondrial cytochrome b (cyt b ) gene indicated that there were at least four distinct populations within the species: Ohio River basin, Upper Mississippi River, Gulf coast, and lower Mississippi River. In particular, the most divergent population was the most recently discovered, from the Elk River, WV, in the Ohio River basin, and it was postulated that this population represents an undescribed, potentially threatened species. However, differentiation observed at a single gene region is generally not considered sufficient evidence to establish taxonomic status. In the present study, nucleotide variation at the mitochondrial control region and a nuclear S7 ribosomal gene intron were compared to provide independent verification of phylogeographic results between individuals collected from the same five disjunct populations previously surveyed. Variation between populations at the control region was substantial (except between Gulf drainages) and was concordant with patterns of sequence divergence from cyt b . Only the Elk River population was resolved as monophyletic based upon nuclear S7, but significant differences based upon Φ ST statistics were observed between most populations. Morphometric data were consistent with molecular data regarding the distinctiveness of the Elk River population. It is proposed that populations of C. asprella consist of at least four distinct population segments, and that the Elk River group likely constitutes a distinct species.

Alabama, Arkansas, Georgia, Louisiana, Mississippi↗
USGS2005

Geostatistical analysis of allele presence patterns among American black bears in eastern North Carolina

Highways are one of the leading causes of wildlife habitat fragmentation and may particularly affect wide-ranging species, such as American black bears (Ursus americanus). We initiated a research project in 2000 to determine potential effects of a 4-lane highway on black bear ecology in Washington County, North Carolina. The research design included a treatment area (highway construction) and a control area and a pre- and post-construction phase. We used data from the pre-construction phase to determine whether we could detect scale dependency or directionality among allele occurrence patterns using geostatistics. Detection of such patterns could provide a powerful tool to measure the effects of landscape fragmentation on gene flow. We sampled DNA from roots of black bear hair at 70 hair-sampling sites on each study area for 7 weeks during fall of 2000. We used microsatellite analysis based on 10 loci to determine unique multi-locus genotypes. We examined all alleles sampled at ???25 sites on each study area and mapped their presence or absence at each hair-sample site. We calculated semivariograms, which measure the strength of statistical correlation as a function of distance, and adjusted them for anisotropy to determine the maximum direction of spatial continuity. We then calculated the mean direction of spatial continuity for all examined alleles. The mean direction of allele frequency variation was 118.3?? (SE = 8.5) on the treatment area and 172.3?? (SE = 6.0) on the control area. Rayleigh's tests showed that these directions differed from random distributions (P = 0.028 and P < 0.001, respectively), indicating consistent directional patterns for the alleles we examined in each area. Despite the small spatial scale of our study (approximately 11,000 ha for each study area), we observed distinct and consistent patterns of allele occurrence, suggesting different directions of gene flow between the study areas. These directions seemed to coincide with the primary orientation of the best habitat areas. Furthermore, the patterns we observed suggest directions of potential source populations beyond the 2 study areas. Indeed, nearby areas classified as core black bear habitat exist in the directions indicated by our analysis. Geostatistical analysis of allele occurrence patterns may provide a useful technique to identify potential barriers to gene flow among bear populations.

Ursus↗
USGS2005

Genetic diversity, kinship analysis, and broodstock management of captive Atlantic sturgeon for population restoration

Captive Atlantic sturgeon Acipenser oxyrinchus considered for use as broodstock in a restoration program were genotyped using nuclear DNA microsatellites and compared to wild collections from the Hudson River, New York (source of parents of the captive sturgeon) and from Albemarle Sound, North Carolina. Because the potential broodfish were the progeny of a small number of parents, maintaining genetic diversity and minimizing inbreeding is essential to a successful breeding and supplementation program. The microsatellite loci used in this analysis generated unique multilocus genotypes for each of 136 Atlantic sturgeon. Analyses indicated significant genetic separation between the New York and North Carolina collections and correctly identified the potential broodstock as a subset of the Hudson River population. Pairwise genetic distance (-In proportion of shared alleles) between half and full siblings in the potential broodfish was as great as 1.386, a value exceeded by only 36% of the sampled broodfish pairs available for mating. Because the current broodstock population does not seem to have deviated far from their ancestral population in the Hudson River, progeny from that broodstock, or the parents themselves, would seem to be genetically suitable for release back into the Hudson River.

American Fisheries Society Symposium↗
USGS2005

Molecular identification of cypripedioid orchids in international trade

Two cypripedioid orchid genera, Paphiopedilum and Phragmipedium, are listed in Appendix I of CITES and are restricted from international trade. Because of their morphological similarity to other genera, however, they may be disguised as belonging to one of the other cypripedioids listed along with other orchids in Appendix II of CITES. Sequence analysis was performed on the internal transcribed spacer region (ITS) of ribosomal DNA of cypripedioid orchids to develop a molecular marker system capable of discriminating among rare species in trade. Molecular analyses concentrated on rare cypripedioid orchids from the genera Paphiopedilum and Phragmipedium, which are known to be poached from the wild and smuggled across international borders disguised as common species. A total of 48 taxa representing two genera {Paphiopedilum, N = 43; Phragmipedium, N = 5) have been sequenced and compared for distinc- tiveness. Phylogenetic analyses clearly distinguish between these two genera and among other cypripedioid genera, with 5-10 fixed nucleotide differences reported between genera. Within a genus, sections of closely related taxa are recoverable in phylogenetic analyses, in most cases, with low sequence divergence within sections. ITS sequences available in GenBank have been aligned with data generated for this project, resulting in a comprehensive sequence library of 151 sequences representing all genera of cypripedioid orchids: 70 Paphiopedilum taxa, 16 Phragmipedium taxa, and 14 Cypripedium taxa, as well as represen- tatives from Selenipedium and the monotypic genus Mexipedium (Phragmipedium) xerophyticum. Addi- tionally, several organelle intron regions have been screened for variation among genera and species. Both the chloroplast řrnS-M and the mitochondrial NAD1 intron regions, which varied between genera in nu- cleotide substitutions and indels, hold promise for increasing ability to distinguish between these orchids. The set of DNA markers examined for this project are diagnostic of these genera, appear to be robust, and are suitable for rapid assay to avoid unnecessary complication in the legitimate trade of orchids listed in CITES Appendix

Selbyana↗
USGS2003

Population structure of Atlantic salmon in Maine with reference to populations from Atlantic Canada

Anadromous Atlantic salmon Salmo salar from 12 rivers in Maine, 3 rivers in New Brunswick, and 2 rivers each in Nova Scotia, Quebec, Newfoundland, and Labrador as well as 2 landlocked strains in Maine ( N = 3,863) were genotyped at 11 microsatellite loci. Fish in the drainages of Maine's Kennebec and Penobscot rivers were genetically similar to those sampled from the 8 rivers recently listed as containing an endangered distinct population segment under the United States' Endangered Species Act. Genetic distance estimates confirm that Maine's Atlantic salmon, both landlocked and anadromous, represent a discrete population unit, genetically as independent from any Canadian population as the Canadian populations are from each other. Within Maine, the anadromous and landlocked populations were statistically distinct. Anadromous Atlantic salmon were more genetically similar among year-classes within rivers than among rivers, as would be expected if the river is the unit of population. The effective number of breeders estimated within each river is larger than the number of adults estimated from samples and redd counts over the 10-year period from 1991 to 2000.

Maine↗
USGS2002

Intraspecific phylogeography of Lasmigona subviridis (Bivalvia: Unionidae): Conservation implications of range discontinuity

A nucleotide sequence analysis of the first internal transcribed spacer region (ITS-1) between the 5.8S and 18S ribosomal DNA genes (640 bp) and cytochrome c oxidase subunit I (COI) of mitochondrial DNA (mtDNA) (576 bp) was conducted for the freshwater bivalve Lasmigona subviridis and three congeners to determine the utility of these regions in identifying phylogeographic and phylogenetic structure. Sequence analysis of the ITS-1 region indicated a zone of discontinuity in the genetic population structure between a group of L. subviridis populations inhabiting the Susquehanna and Potomac Rivers and more southern populations. Moreover, haplotype patterns resulting from variation in the COI region suggested an absence of gene exchange between tributaries within two different river drainages, as well as between adjacent rivers systems. The authors recommend that the northern and southern populations, which are reproductively isolated and constitute evolutionarily significant lineages, be managed as separate conservation units. Results from the COI region suggest that, in some cases, unionid relocations should be avoided between tributaries of the same drainage because these populations may have been reproductively isolated for thousands of generations. Therefore, unionid bivalves distributed among discontinuous habitats (e.g. Atlantic slope drainages) potentially should be considered evolutionarily distinct. The DNA sequence divergences observed in the nuclear and mtDNA regions among the Lasmigona species were congruent, although the level of divergence in the COI region was up to three times greater. The genus Lasmigona , as represented by the four species surveyed in this study, may not be monophyletic.

Delaware, Maryland, North Carolina, Pennsylvania, ↗
USGS2001

Microsatellite DNA markers for the study of Allegheny woodrat (Neotoma magister) populations and cross-species amplification the genus Neotoma

The Allegheny woodrat ( Neotoma magister ) is a colony-forming murid rodent inhabiting rock outcrops, cliff and talus slopes, and caves within the Central and Southern Appalachians of North America ( Hall 1981 ). The species is currently considered threatened, endangered, or at risk throughout its range purportedly due to the direct or indirect effects of human-fragmented landscapes. Currently, Allegheny woodrat management units are arbitrarily defined as metapopulations that roughly constitute contiguous rock outcrops interconnected by forested habitat. To strengthen long-term population stability and reduce the need for further protection through the regulatory process, an effective conservation programme for this at-risk species will require unambiguous, objectively defined units of management that discern evolutionarily important lineages ( Avise 1994 ). A prerequisite in the development of management strategies that recognize and maintain significant woodrat lineages is a thorough understanding of the levels of gene exchange among geographically proximate and distal populations. However, no information exists on population structure, levels of gene flow, or relatedness among Allegheny woodrat populations or colonies. As an initial step towards addressing this research need, we have developed a suite of polymorphic microsatellite DNA markers for this species. In this paper, we describe the isolation and characterization of these markers and demonstrate their suitability in amplifying putatively homologous products in seven additional Neotoma species.

Alabama, Georgia, Kentucky, Mississippi, North Car↗
USGS2001

Fine-scale population structure Atlantic salmon from Maine's Penobscot River drainage

We report a survey of microsatellite DNA variation in Atlantic salmon from the unimpounded lower reaches of Maine's Penobscot River. Our analysis indicates that Atlantic salmon in the Penobscot River are distinct from other populations that have little or no history of human-mediated repopulation, including two of its tributaries, Cove Brook and Kenduskeag Stream, another Maine river, the Ducktrap, and Canada's Miramichi and Gander rivers. Significant heterogeneity was detected in allele frequency among all three subpopulations sampled in the Penobscot drainage. The high resolution of the 12-locussuite was quantified using maximum likelihood assignment tests, which correctly identified the source of 90.4–96.1% of individuals from within the Penobscot drainage. Current populations are clearly isolated from each other, however we are unable to determine from the present data whether the populations in Cove Brook and Kenduskeag Stream are recently diverged from populations stocked into the Penobscot River over the last century, or are aboriginal in origin. The degree of population structure identified in the Penobscot drainage is noteworthy in light of its lengthy history of systematic restocking, the geographic proximity of the subpopulations, and the extent of the differentiation. Similar population structure on this extremely limited geographic scale could exist among Atlantic salmon runs elsewhere in Maine and throughout the species' range and should be taken into account for future management decisions.

Maine↗