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At least 181 records · Page 10Linked to original sources

Efficient genotyping with backwards compatibility: Converting a legacy microsatellite panel for muskellunge (Esox masquinongy) to genotyping-by-sequencing chemistry

Microsatellites have been a staple of population genetics research for over three decades, and many large datasets have been generated with these markers. Microsatellites have been used, for example, to conduct genetic monitoring and construct large multigeneration pedigrees as well as genotype thousands of individuals from a given species to create high-resolution baselines of spatial genetic structure. However, the capillary electrophoresis (CE) approach used to genotype microsatellites is inefficient compared to newer genotyping-by-sequencing (GBS) approaches, and researchers have begun transitioning away from CE. Backward compatibility between GBS and CE would facilitate a seamless transition to a more efficient chemistry, while ensuring that research based on CE panels could continue. Here, we explore the feasibility of converting a legacy panel of 15 microsatellites developed for muskellunge ( Esox masquinongy ) from CE to GBS chemistry. Muskellunge are an important sportfish in the Great Lakes region, and the existing microsatellite panel has been used to genotype thousands of samples to develop a region-wide baseline of genetic structure. We successfully converted all 15 microsatellites to GBS chemistry. GBS produced high genotyping rates (98%) and had high concordance with CE microsatellite genotypes (99%). Conversion to GBS required redesign of some primers and pairs to shorten amplicon length and adjust melting temperatures, optimization of primer concentrations, and comparisons with CE genotypes to optimize GBS genotyping parameters; however, none of these steps were especially onerous. Our results demonstrate that it is highly feasible to convert legacy CE panels to GBS, ensuring seamless continuation of important, often long-term research.

Conservation Genetics Resources

Complex patterns of genetic and morphological differentiation in the Smallmouth Bass subspecies (Micropterus dolomieu dolomieu and M. d. velox) of the Central Interior Highlands

Due to geologic processes and recent anthropogenic introductions, patterns of genetic and morphological diversity within the Smallmouth Bass ( Micropterus dolomieu ), which are endemic to the central and eastern United States (USA), are poorly understood. We assessed genetic and morphological differentiation between the widespread Northern Smallmouth Bass ( M. d. dolomieu ) and the more restricted Neosho Smallmouth Bass ( M. d. velox ) where their ranges meet in the Central Interior Highlands ecoregion (CIH). Data from 14 microsatellite loci were used to conduct S TRUCTURE and principal components analyses to evaluate diversity across populations and screen for hybridization with sympatric Spotted Bass ( M. punctulatus ). We also tested for morphological differences using five morphometric traits and one meristic trait. We found support for three genetic clusters corresponding to previously described taxonomic variation; five clusters largely corresponding to river systems; and nine clusters representing hierarchical population structure within both ranges. We found evidence of a unique genetic cluster in tributaries of the White River within the Northern Smallmouth Bass range and admixture between the subspecies throughout the Neosho range. We also found evidence of morphological differentiation between subspecies; Neosho Smallmouth Bass exhibited larger head length than Northern Smallmouth Bass relative to total length, and there was a significant interaction of subspecies and orbital length, possibly indicating differential growth patterns between subspecies. Our results reveal multiple levels of divergence, suggesting the CIH harbors greater and more complex Smallmouth Bass diversity than previously thought.

Arkansas, Illinois, Kansas, Mississippi, Missouri,

Molecular and phenotypic diversity in Chionactis occipitalis (Western Shovel-nosed Snake), with emphasis on the status of C. o. klauberi (Tucson Shovel-nosed Snake).

Chionactis occipitalis (Western Shovel-nosed Snake) is a small colubrid snake inhabiting the arid regions of the Mojave, Sonoran, and Colorado deserts. Morphological assessments of taxonomy currently recognize four subspecies. However, these taxonomic proposals were largely based on weak morphological differentiation and inadequate geographic sampling. Our goal was to explore evolutionary relationships and boundaries among subspecies of C. occipitalis, with particular focus on individuals within the known range of C. o. klauberi (Tucson Shovel-nosed snake). Population sizes and range for C. o. klauberi have declined over the last 25 years due to habitat alteration and loss prompting a petition to list this subspecies as endangered. We examined the phylogeography, population structure, and subspecific taxonomy of C. occipitalis across its geographic range with genetic analysis of 1100 bases of mitochondrial DNA sequence and reanalysis of 14 morphological characters from 1543 museum specimens. We estimated the species gene phylogeny from 81 snakes using Bayesian inference and explored possible factors influencing genetic variation using landscape genetic analyses. Phylogenetic and population genetic analyses reveal genetic isolation and independent evolutionary trajectories for two primary clades. Our data indicate that diversification between these clades has developed as a result of both historical vicariance and environmental isolating mechanisms. Thus these two clades likely comprise 'evolutionary significant units' (ESUs). Neither molecular nor morphological data are concordant with the traditional C. occipitalis subspecies taxonomy. Mitochondrial sequences suggest specimens recognized as C. o. klauberi are embedded in a larger geographic clade whose range has expanded from western Arizona populations, and these data are concordant with clinal longitudinal variation in morphology. ?? 2007 Springer Science+Business Media B.V.

Conservation Genetics

Novel microsatellite loci for studies of Thamnophis Gartersnake genetic identity and hybridization

Butler’s Gartersnakes (BGS; Thamnophis butleri ) are confined to open and semi-open canopy wetlands and adjacent uplands, habitats under threat of development in Wisconsin. To address issues of species identity and putative hybridization with congeneric snakes, a suite of 18 microsatellite loci capable of cross-species amplification of Plains Gartersnakes ( T. radix ) and Common Gartersnakes ( T. sirtalis ) was developed. All loci were polymorphic in BGS with mean number of alleles per locus of 16.11 (range = 3–41) and mean observed heterozygosity of 0.659 (range = 0.311–0.978). Loci amplified efficiently in the congeneric species with high levels of intra- and inter-specific variation. These loci will aid ongoing efforts to effectively identify and manage BGS in Wisconsin.

Conservation Genetics Resources

Genetic structure of Florida green turtle rookeries as indicated by mitochondrial DNA control region sequences

Green turtle ( Chelonia mydas ) nesting has increased dramatically in Florida over the past two decades, ranking the Florida nesting aggregation among the largest in the Greater Caribbean region. Individual beaches that comprise several hundred kilometers of Florida’s east coast and Keys support tens to thousands of nests annually. These beaches encompass natural to highly developed habitats, and the degree of demographic partitioning among rookeries was previously unresolved. We characterized the genetic structure of ten Florida rookeries from Cape Canaveral to the Dry Tortugas through analysis of 817 base pair mitochondrial DNA ( mtDNA ) control region sequences from 485 nesting turtles. Two common haplotypes, CM-A1.1 and CM-A3.1, accounted for 87 % of samples, and the haplotype frequencies were strongly partitioned by latitude along Florida’s Atlantic coast. Most genetic structure occurred between rookeries on either side of an apparent genetic break in the vicinity of the St. Lucie Inlet that separates Hutchinson Island and Jupiter Island, representing the finest scale at which mtDNA structure has been documented in marine turtle rookeries. Florida and Caribbean scale analyses of population structure support recognition of at least two management units: central eastern Florida and southern Florida. More thorough sampling and deeper sequencing are necessary to better characterize connectivity among Florida green turtle rookeries as well as between the Florida nesting aggregation and others in the Greater Caribbean region.

Florida

Genetic discontinuity among regional populations of Lophelia perfusa in the North Atlantic Ocean

Knowledge of the degree to which populations are connected through larval dispersal is imperative to effective management, yet little is known about larval dispersal ability or population connectivity in Lophelia pertusa, the dominant framework-forming coral on the continental slope in the North Atlantic Ocean. Using nine microsatellite DNA markers, we assessed the spatial scale and pattern of genetic connectivity across a large portion of the range of L. pertusa in the North Atlantic Ocean. A Bayesian modeling approach found four distinct genetic groupings corresponding to ocean regions: Gulf of Mexico, coastal southeastern U.S., New England Seamounts, and eastern North Atlantic Ocean. An isolation-by-distance pattern was supported across the study area. Estimates of pairwise population differentiation were greatest with the deepest populations, the New England Seamounts (average FST = 0.156). Differentiation was intermediate with the eastern North Atlantic populations (FST = 0.085), and smallest between southeastern U.S. and Gulf of Mexico populations (FST = 0.019), with evidence of admixture off the southeastern Florida peninsula. Connectivity across larger geographic distances within regions suggests that some larvae are broadly dispersed. Heterozygote deficiencies were detected within the majority of localities suggesting deviation from random mating. Gene flow between ocean regions appears restricted, thus, the most effective management scheme for L. pertusa involves regional reserve networks

Conservation Genetics

Quantitative PCR detection of endangered diamond darter Crystallaria Cincotta in environmental DNA: Employing locked nucleic acids and blocking probe for specificity

This study presents a quantitative PCR (qPCR) assay for the detection of the endangered diamond darter Crystallaria cincotta from environmental DNA (eDNA) in water samples. The assay design is based on an alignment of mitochondrial cytochrome b DNA sequences from 58 individuals representing 25 percid species. Leveraging genetic differences, a species-specific qPCR assay was designed, incorporating alocked nucleic acid (LNA)-enriched probe and a secondary blocking probe to enhance specificity. The assay targets a 93-base pair fragment that includes a diagnostic single nucleotide polymorphism in the probe region; combined with multiple primer mismatches, this provides specificity for distinguishing C. cincotta from other sympatric percid species. Specificity was validated by testing genomic DNA from 16 percid species and synthetic templates, confirming no cross-reactivity. Performance metrics, including the standard curve, qPCR efficiency, limit of detection, and limit of quantification, are reported. The qPCR assay exhibited sufficient sensitivity to detect C. cincotta eDNA in environmental water samples collected from occupied riverine habitats. This study illustrates the effectiveness of LNA-enriched and blocking probes in developing species-specific qPCR assays for eDNA applications, demonstrating their utility in accurately distinguishing closely related species within diverse fish communities.

Conservation Genetics Resources

Rapid isolation of microsatellite DNAs and identification of polymorphic mitochondrial DNA regions in the fish rotan (Perccottus glenii) invading European Russia

Human-mediated translocations and subsequent large-scale colonization by the invasive fish rotan (Perccottus glenii Dybowski, 1877; Perciformes, Odontobutidae), also known as Amur or Chinese sleeper, has resulted in dramatic transformations of small lentic ecosystems. However, no detailed genetic information exists on population structure, levels of effective movement, or relatedness among geographic populations of P. glenii within the European part of the range. We used massively parallel genomic DNA shotgun sequencing on the semiconductor-based Ion Torrent Personal Genome Machine (PGM) sequencing platform to identify nuclear microsatellite and mitochondrial DNA sequences in P. glenii from European Russia. Here we describe the characterization of nine nuclear microsatellite loci, ascertain levels of allelic diversity, heterozygosity, and demographic status of P. glenii collected from Ilev, Russia, one of several initial introduction points in European Russia. In addition, we mapped sequence reads to the complete P. glenii mitochondrial DNA sequence to identify polymorphic regions. Nuclear microsatellite markers developed for P. glenii yielded sufficient genetic diversity to: (1) produce unique multilocus genotypes; (2) elucidate structure among geographic populations; and (3) provide unique perspectives for analysis of population sizes and historical demographics. Among 4.9 million filtered P. glenii Ion Torrent PGM sequence reads, 11,304 mapped to the mitochondrial genome (NC_020350). This resulted in 100 % coverage of this genome to a mean coverage depth of 102X. A total of 130 variable sites were observed between the publicly available genome from China and the studied composite mitochondrial genome. Among these, 82 were diagnostic and monomorphic between the mitochondrial genomes and distributed among 15 genome regions. The polymorphic sites (N = 48) were distributed among 11 mitochondrial genome regions. Our results also indicate that sequence reads generated from two three-hour runs on the Ion Torrent PGM can generate a sufficient number of nuclear and mitochondrial markers to improve understanding of the evolutionary and ecological dynamics of non-model and in particular, invasive species.

European Russia

Identification of metapopulation dynamics among Northern Goshawks of the Alexander Archipelago, Alaska, and Coastal British Columbia

Northern Goshawks occupying the Alexander Archipelago, Alaska, and coastal British Columbia nest primarily in old-growth and mature forest, which results in spatial heterogeneity in the distribution of individuals across the landscape. We used microsatellite and mitochondrial data to infer genetic structure, gene flow, and fluctuations in population demography through evolutionary time. Patterns in the genetic signatures were used to assess predictions associated with the three population models: panmixia, metapopulation, and isolated populations. Population genetic structure was observed along with asymmetry in gene flow estimates that changed directionality at different temporal scales, consistent with metapopulation model predictions. Therefore, Northern Goshawk assemblages located in the Alexander Archipelago and coastal British Columbia interact through a metapopulation framework, though they may not fit the classic model of a metapopulation. Long-term population sources (coastal mainland British Columbia) and sinks (Revillagigedo and Vancouver islands) were identified. However, there was no trend through evolutionary time in the directionality of dispersal among the remaining assemblages, suggestive of a rescue-effect dynamic. Admiralty, Douglas, and Chichagof island complex appears to be an evolutionarily recent source population in the Alexander Archipelago. In addition, Kupreanof island complex and Kispiox Forest District populations have high dispersal rates to populations in close geographic proximity and potentially serve as local source populations. Metapopulation dynamics occurring in the Alexander Archipelago and coastal British Columbia by Northern Goshawks highlight the importance of both occupied and unoccupied habitats to long-term population persistence of goshawks in this region.

Alaska, British Columbia

Spatial genetic structure of bristle-thighed curlews (Numenius tahitiensis): Breeding area differentiation not reflected on the non-breeding grounds

Migratory birds occupy geographically and ecologically disparate areas during their annual cycle with conditions on breeding and non-breeding grounds playing separate and important roles in population dynamics. We used data from nuclear microsatellite and mitochondrial DNA control region loci to assess the breeding and non-breeding spatial genetic structure of a transoceanic migrant shorebird, the bristle-thighed curlew. We found spatial variance in the distribution of allelic and haplotypic frequencies between the curlew's two breeding areas in Alaska but did not observe this spatial structure throughout its non-breeding range on low-lying tropical and subtropical islands in the Central Pacific (Oceania). This suggests that the two breeding populations do not spatially segregate during the non-breeding season. Lack of migratory connectivity is likely attributable to the species' behavior, as bristle-thighed curlews exhibit differential timing of migration and some individuals move among islands during non-breeding months. Given the detrimental impact of many past and current human activities on island ecosystems, admixture of breeding populations in Oceania may render the bristle-thighed curlew less vulnerable to perturbations there, as neither breeding population will be disproportionally affected by local habitat losses or by stochastic events. Furthermore, lack of migratory connectivity may enable bristle-thighed curlews to respond to changing island ecosystems by altering their non-breeding distribution. However, availability of suitable non-breeding habitat for curlews in Oceania is increasingly limited on both low-lying and high islands by habitat loss, sea level rise, and invasive mammalian predators that pose a threat to flightless and flight-compromised curlews during the molting period.

Conservation Genetics

Population structure and genetic stock identification in southeastern United States loggerhead sea turtles (Caretta caretta) using genome-wide SNPs

Characterizing the genetic structure and connectivity between populations of endangered species can be used to inform management actions. In vagile species with high gene flow or recently established populations, such characterizations can be difficult to undertake using traditional genetic markers, and genetic stock identification (GSI) may be confounded by allele-sharing between populations. Loggerhead sea turtles ( Caretta caretta ) in the southeastern United States comprise seven management units (MUs) based on female philopatry inferred via mitochondrial DNA sequences, yet nuclear microsatellite data do not reflect divergence. Further, loci for accurate GSI are not currently known. To address this, we generated genome-wide single nucleotide polymorphism (SNP) data from 146 females nesting at individual sites representative of each southeastern United States MU. We found weak (F ST =0.001–0.003) but significant divergence among all MUs, with more notable divergence between the Gulf Coast and Atlantic Ocean MUs, and amongst the Atlantic Ocean MUs. We then used an iterative leave-one-out approach to identify candidate loci for GSI. This approach identified loci that could assign individuals to natal ocean basins (i.e., to the Gulf Coast or to the Atlantic Ocean), and to individual MUs within the Atlantic Ocean, with high (≥90%) success and accuracy. Analyses of genome-wide SNPs refined our understanding of the magnitude and scale of population connectivity in loggerhead turtles in the southeastern United States, and provided a foundation for the development of SNP panels for accurate, fine-scale GSI in sea turtles.

Alabama, Florida, Georgia

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

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

The impact of time and field conditions on brown bear ( Ursus arctos ) faecal DNA amplification

To establish longevity of faecal DNA samples under varying summer field conditions, we collected 53 faeces from captive brown bears ( Ursus arctos ) on a restricted vegetation diet. Each faeces was divided, and one half was placed on a warm, dry field site while the other half was placed on a cool, wet field site on Moscow Mountain, Idaho, USA. Temperature, relative humidity, and dew point data were collected on each site, and faeces were sampled for DNA extraction at <1, 3, 6, 14, 30, 45, and 60 days. Faecal DNA sample viability was assessed by attempting PCR amplification of a mitochondrial DNA (mtDNA) locus (???150 bp) and a nuclear DNA (nDNA) microsatellite locus (180-200 bp). Time in the field, temperature, and dew point impacted mtDNA and nDNA amplification success with the greatest drop in success rates occurring between 1 and 3 days. In addition, genotyping errors significantly increased over time at both field sites. Based on these results, we recommend collecting samples at frequent transect intervals and focusing sampling efforts during drier portions of the year when possible. ?? 2007 Springer Science+Business Media, Inc.

Wyoming

Diversity, origins and virulence of Avipoxviruses in Hawaiian Forest Birds

We cultured avian pox (Avipoxvirus spp.) from lesions collected on Hawai'i, Maui, Moloka'i, and 'Oahu in the Hawaiian Islands from 15 native or non-native birds representing three avian orders. Phylogenetic analysis of a 538 bp fragment of the gene encoding the virus 4b core polypeptide revealed two distinct variant clusters, with sequences from chickens (fowlpox) forming a third distinct basal cluster. Pox isolates from one of these two clusters appear closely related to canarypox and other passerine pox viruses, while the second appears more specific to Hawai'i. There was no evidence that birds were infected simultaneously with multiple pox virus variants based on evaluation of multiples clones from four individuals. No obvious temporal or geographic associations were observed and strict host specificity was not apparent among the 4b-defined field isolates. We amplified a 116 bp 4b core protein gene fragment from an 'Elepaio (Chasiempis sandwichensis) collected in 1900 on Hawai'i Island that clustered closely with the second of the two variants, suggesting that this variant has been in Hawai'i for at least 100 years. The high variation detected between the three 4b clusters provides evidence for multiple, likely independent introductions, and does not support the hypothesis of infection of native species through introduction of infected fowl. Preliminary experimental infections in native Hawai'i 'Amakihi (Hemignathus virens) suggest that the 4b-defined variants may be biologically distinct, with one variant appearing more virulent. These pox viruses may interact with avian malaria (Plasmodium relictum), another introduced pathogen in Hawaiian forest bird populations, through modulation of host immune responses. ?? 2007 Springer Science+Business Media B.V.

Conservation Genetics

Uncloaking a cryptic, threatened rail with molecular markers: origins, connectivity and demography of a recently-discovered population

The threatened California Black Rail lives under dense marsh vegetation, is rarely observed, flies weakly and has a highly disjunct distribution. The largest population of rails is found in 8&ndash;10 large wetlands in San Francisco Bay (SF Bay), but a population was recently discovered in the foothills of the Sierra Nevada Mountains (Foothills), within a wetland network comprised of over 200 small marshes. Using microsatellite and mitochondrial analyses, our objectives were to determine the origins, connectivity and demography of this recently-discovered population. Analyses of individuals from the Foothills ( n = 31), SF Bay ( n = 31), the Imperial Valley ( n = 6) and the East Coast ( n = 3), combined with rigorous power evaluations, provided valuable insights into past history and current dynamics of the species in Northern California that challenge conventional wisdom about the species. The Foothills and SF Bay populations have diverged strongly from the Imperial Valley population, even more strongly than from individuals of the East Coast subspecies. The data also suggest a historical presence of the species in the Foothills. The SF Bay and Foothills populations had similar estimated effective population size over the areas sampled and appeared linked by a strongly asymmetrical migration pattern, with a greater probability of movement from the Foothills to SF Bay than vice versa. Random mating was inferred in the Foothills, but local substructure among marshes and inbreeding were detected in SF Bay, suggesting different dispersal patterns within each location. The unexpected dimensions of Black Rail demography and population structure suggested by these analyses and their potential importance for management are discussed.

California

The impact of small irrigation diversion dams on the recent migration rates of steelhead and redband trout ( Oncorhynchus mykiss )

Barriers to migration are numerous in stream environments and can occur from anthropogenic activities (such as dams and culverts) or natural processes (such as log jams or dams constructed by beaver ( Castor canadensis )). Identification of barriers can be difficult when obstructions are temporary or incomplete providing passage periodically. We examine the effect of several small irrigation diversion dams on the recent migration rates of steelhead ( Oncorhynchus mykiss ) in three tributaries to the Methow River, Washington. The three basins had different recent migration patterns: Beaver Creek did not have any recent migration between sites, Libby Creek had two-way migration between sites and Gold Creek had downstream migration between sites. Sites with migration were significantly different from sites without migration in distance, number of obstructions, obstruction height to depth ratio and maximum stream gradient. When comparing the sites without migration in Beaver Creek to the sites with migration in Libby and Gold creeks, the number of obstructions was the only significant variable. Multinomial logistic regression identified obstruction height to depth ratio and maximum stream gradient as the best fitting model to predict the level of migration among sites. Small irrigation diversion dams were limiting population interactions in Beaver Creek and collectively blocking steelhead migration into the stream. Variables related to stream resistance (gradient, obstruction number and obstruction height to depth ratio) were better predictors of recent migration rates than distance, and can provide important insight into migration and population demographic processes in lotic species.

Washington

Speciation with gene flow in a narrow endemic West Virginia cave salamander (Gyrinophilus subterraneus)

Due to their limited geographic distributions and specialized ecologies, cave species are often highly endemic and can be especially vulnerable to habitat degradation within and surrounding the cave systems they inhabit. We investigated the evolutionary history of the West Virginia Spring Salamander ( Gyrinophilus subterraneus) , estimated the population trend from historic and current survey data, and assessed the current potential for water quality threats to the cave habitat. Our genomic data (mtDNA sequence and ddRADseq-derived SNPs) reveal two, distinct evolutionary lineages within General Davis Cave corresponding to G. subterraneus and its widely distributed sister species, Gyrinophilus porphyriticus , that are also differentiable based on morphological traits. Genomic models of evolutionary history strongly support asymmetric and continuous gene flow between the two lineages, and hybrid classification analyses identify only parental and first generation cross (F1) progeny. Collectively, these results point to a rare case of sympatric speciation occurring within the cave, leading to strong support for continuing to recognize G. subterraneus as a distinct and unique species. Due to its specialized habitat requirements, the complete distribution of G. subterraneus is unresolved, but using survey data in its type locality (and currently the only known occupied site), we find that the population within General Davis Cave has possibly declined over the last 45 years. Finally, our measures of cave and surface stream water quality did not reveal evidence of water quality impairment and provide important baselines for future monitoring. In addition, our unexpected finding of a hybrid zone and partial reproductive isolation between G. subterraneus and G. porphyriticus warrants further attention to better understand the evolutionary and conservation implications of occasional hybridization between the species.

West Virginia