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At least 397 records · Page 22Linked to original sources

Defining the need for genetic stock assignment when describing stock demographics and dynamics: An example using Lake Whitefish in Lake Michigan

Genetic stock assignment is not routinely used when describing the dynamics and demographics of individual stocks supporting mixed-stock fisheries, and capture location and timing are often used as alternative assignment methods. However, variation in stock demographics and dynamics may not be accounted for if stock assignments based on capture location or timing do not accurately reflect genetic assignments. We used Lake Whitefish Coregonus clupeaformis in Lake Michigan as a model fishery to determine whether stock mixing could undermine efforts to describe stock status when using October capture location as a proxy for genetic stock assignment. Accuracy of stock assignments based on October capture location ranged from 54% to 100% among management zones. Metrics describing length and age distributions, weight at length, fecundity, and growth varied among genetic stocks. Stock-specific metrics were typically similar between stock assignment methods (capture location versus genetics) because only one or two genetic stocks were collected in most locations and the majority of those fish were from spatially proximal stocks with similar metrics. However, more extensive mixing of Lake Whitefish stocks has been documented; thus, using capture location for stock assignment could result in incorrect conclusions regarding stock status and harvest management depending on stock composition. Ambiguity in genetic stock assignments was a problem in two management zones, where between 23% and 42% of Lake Whitefish did not assign to a specific stock with a probability of at least 0.70. In the future, using genomic techniques rather than microsatellites may provide different conclusions regarding genetic stock structure; these differences could affect the accuracy of using capture location for stock assignment. Use of capture location as a proxy for genetic stock assignment may not be warranted for all mixed-stock fisheries but may be appropriate when stock mixing is limited or is restricted to stocks with consistently similar characteristics.

Michigan, Wisconsin↗

Polyphosphate present in DNA preparations from fungal species of Collectotrichum inhibits restriction endonucleases and other enzymes

During the development of a procedure for the isolation of total genomic DNA from filamentous fungi (Rodriguez, R. J., and Yoder, 0. C., Exp. Mycol. 15, 232-242, 1991) a cell fraction was isolated which inhibited the digestion of DNA by restriction enzymes. After elimination of DNA, RNA, proteins, and lipids, the active compound was purified by gel filtration to yield a single fraction capable of complete inhibition of restriction enzyme activity. The inhibitor did not absorb uv light above 220 nm, and was resistant to alkali and acid at 25°C and to temperatures as high as 100°C. More extensive analyses demonstrated that the inhibitor was also capable of inhibiting T4 DNA ligase and Taq I DNA polymerase, but not DNase or RNase. Chemical analyses indicated that the inhibitor was devoid of carbohydrates, proteins, lipids, and nucleic acids but rich in phosphorus. A combination of nuclear magnetic resonance, metachromatic shift of toluidine blue, and gel filtration indicated that the inhibitor was a polyphosphate (polyP) containing approximately 60 phosphate molecules. The mechanism of inhibition appeared to involve complexing of polyP to the enzymatic proteins. All species of Colletotrichum analyzed produced polyP equivalent in chain length and concentration. A modification to the original DNA extraction procedure is described which eliminates polyP and reduces the time necessary to obtain DNA of sufficient purity for restriction enzyme digestion and Taq I polymerase amplification.

Analytical Biochemistry↗

International importance of Percids: Summary and looking forward

Research presented in the preceding chapters emphasizes recent advancements in the research, management, and aquaculture of Walleye, Sauger, and Yellow Perch in North America. These percid fishes, along with the European Perch and Pikeperch, are economically and ecologically important fishes in their native geographic range. Advances in techniques to evaluate current habitat and predict future habitat conditions provide managers with detailed baseline information and biophysical models useful for evaluating adaptive management practices. Current habitat use and movement assessments have improved substantially with technological advancements in acoustic tags and extensive receiver array networks, which, combined with genetic and genomic tools, are improving percid stock assessments and management. Advances in percid aquaculture techniques have improved growth, survival, and disease resistance, enhancing percid stocking efforts and the production of marketable fish. The exchange of information between researchers and managers will continue to advance techniques of percid management for commercial and recreational exploitation and improve aquaculture practices to provide a lucrative commercial aquaculture industry.

Book chapter↗

Adaptation and survival of plants in high stress habitats via fungal endophyte conferred stress tolerance

From the Arctic to the Antarctic, plants thrive in diverse habitats that impose different levels of adaptive pressures depending on the type and degree of biotic and abiotic stresses inherent to each habitat (Stevens, 1989). At any particular location, the abundance and distribution of individual plant species vary tremendously and is theorized to be based on the ability to tolerate a wide range of edaphic conditions and habitat-specific stresses (Pianka, 1966). The ability of individual plant species to thrive in diverse habitats is commonly referred to as phenotypic plasticity and is thought to involve adaptations based on changes in the plant genome (Givnish, 2002; Pan et al., 2006; Robe and Griffiths, 2000; Schurr et al., 2006). Habitats that impose high levels of abiotic stress are typically colonized with fewer plant species compared to habitats imposing low levels of stress. Moreover, high stress habitats have decreased levels of plant abundance compared to low stress habitats even though these habitats may occur in close proximity to one another (Perelman et al., 2007). This is particularly interesting because all plants are known to perceive, transmit signals, and respond to abiotic stresses such as drought, heat, and salinity (Bartels and Sunkar, 2005; Bohnert et al., 1995). Although there has been extensive research performed to determine the genetic, molecular, and physiological bases of how plants respond to and tolerate stress, the nature of plant adaptation to high stress habitats remains unresolved (Leone et al., 2003; Maggio et al., 2003; Tuberosa et al., 2003). However, recent evidence indicates that a ubiquitous aspect of plant biology (fungal symbiosis) is involved in the adaptation and survival of at least some plants in high stress habitats (Rodriguez et al., 2008).

Book chapter↗

Identification and regulatory analysis of rainbow trout tapasin and tapasin-related genes

Tapasin (TAPBP) is a key member of MHC class Ia antigen-loading complexes, bridging the class Ia molecule to the transporter associated with antigen presentation (TAP). As part of an ongoing study of MHC genomics in rainbow trout, we have identified two rainbow trout TAPBP genes (Onmy-TAPBP.a and .b) and a similar but distinct TAPBP-related gene (Onmy-TAPBP-R) that had previously only been described in mammals. Physical and genetic mapping indicate that Onmy-TAPBP.a is on chromosome 18 in the MHC class Ia region and that Onmy-TAPBP.b resides on chromosome 14 in the MHC class Ib region. There are also at least two copies of TAPBP-R, Onmy-TAPBP-R.a and Onmy-TAPBP-R.b, located on chromosomes 2 and 3, respectively. Due to the central role of TAPBP expression during acute viral infection, we have characterized the transcriptional profile and regulatory regions for both Onmy-TAPBP and Onmy-TAPBP-R. Transcription of both genes increased during acute infection with infectious hematapoeitic necrosis virus (IHNV) in a fashion indicative of interferon-mediated regulation. Promoter-reporter assays in STE-137 cells demonstrate that the trout TAPBP and TAPBP-R promoters respond to interferon regulatory factors, Onmy-IRF1 and Onmy-IRF2. Overall, TAPBP is expressed at higher levels than TAPBP-R in nai??ve tissues and TAPBP transcription is more responsive to viral infection and IRF1 and 2 binding. ?? Springer-Verlag 2006.

Immunogenetics↗

Selection, trans-species polymorphism, and locus identification of major histocompatibility complex class IIβ alleles of New World ranid frogs

Genes encoded by the major histocompatibility complex (MHC) play key roles in the vertebrate immune system. However, our understanding of the evolutionary processes and underlying genetic mechanisms shaping these genes is limited in many taxa, including amphibians, a group currently impacted by emerging infectious diseases. To further elucidate the evolution of the MHC in frogs (anurans) and develop tools for population genetics, we surveyed allelic diversity of the MHC class II ??1 domain in both genomic and complementary DNA of seven New World species in the genus Rana (Lithobates). To assign locus affiliation to our alleles, we used a "gene walking" technique to obtain intron 2 sequences that flanked MHC class II?? exon 2. Two distinct intron sequences were recovered, suggesting the presence of at least two class II?? loci in Rana. We designed a primer pair that successfully amplified an orthologous locus from all seven Rana species. In total, we recovered 13 alleles and documented trans-species polymorphism for four of the alleles. We also found quantitative evidence of selection acting on amino acid residues that are putatively involved in peptide binding and structural stability of the ??1 domain of anurans. Our results indicated that primer mismatch can result in polymerase chain reaction (PCR) bias, which influences the number of alleles that are recovered. Using a single locus may minimize PCR bias caused by primer mismatch, and the gene walking technique was an effective approach for generating single-copy orthologous markers necessary for future studies of MHC allelic variation in natural amphibian populations. ?? 2010 Springer-Verlag.

Immunogenetics↗

Arrested development of the myxozoan parasite, Myxobolus cerebralis, in certain populations of mitochondrial 16S lineage III Tubifex tubifex

Laboratory populations of Tubifex tubifex from mitochondrial (mt)16S ribosomal DNA (rDNA) lineage III were generated from single cocoons of adult worms releasing the triactinomyxon stages (TAMs) of the myxozoan parasite, Myxobolus cerebralis. Subsequent worm populations from these cocoons, referred to as clonal lines, were tested for susceptibility to infection with the myxospore stages of M. cerebralis. Development and release of TAMs occurred in five clonal lines, while four clonal lines showed immature parasitic forms that were not expelled from the worm (non-TAM producers). Oligochaetes from TAM- and non-TAM-producing clonal lines were confirmed as lineage III based on mt16S rDNA and internal transcribed spacer region 1 (ITS1) sequences, but these genes did not differentiate these phenotypes. In contrast, random amplified polymorphic DNA analyses of genomic DNA demonstrated unique banding patterns that distinguished the phenotypes. Cohabitation of parasite-exposed TAM- and non-TAM-producing phenotypes showed an overall decrease in expected TAM production compared to the same exposure dose of the TAM-producing phenotype without cohabitation. These studies suggest that differences in susceptibility to parasite infection can occur in genetically similar T. tubifex populations, and their coexistence may affect overall M. cerebralis production, a factor that may influence the severity of whirling disease in wild trout populations. ?? 2007 Springer-Verlag.

Parasitology Research↗

Diverse novel and avian-associated viruses in the ileal viromes of northern mockingbird (Mimus polyglottos)

Viruses are the most abundant and diverse organisms on Earth, though only a small portion cause disease. Understanding viral diversity is key to understanding and predicting pathogen emergence and zoonotic spillover. Here, we use meta-transcriptomic sequencing to examine the viral communities in the ileum of 25 Northern Mockingbirds ( Mimus polyglottos ) from various locations across Texas. We assembled high-quality genomes of 43 viral species (40 species identified to 13 families, one to kingdom, and two to realm), 38 of which were novel. They tentatively represent avian- (n = 3), arthropod- (n = 21), plant- (n = 5) and fungi- (n = 4) associated, or other (n = 10) viruses. The arthropod-associated Dicistroviridae family was the most dominant, comprising known and potentially new species. Of potential epidemiological importance were three novel and avian-associated viruses: members of the families Hepeviridae and Picornaviridae , and a new Matryoshka RNA virus. The Matryoshka RNA virus 8 (MaRNAV-8) is sister to other Matryoshka RNA viruses, and its co-occurrence with haemosporida further supports the nested virus-parasite-vector-vertebrate host relationship of this group of viruses, with potential implications for parasite evolution, fitness and load and vector competence. The Picornaviridae virus is a member of an avian hepatovirus clade, found nested within a clade containing both the mammalian pathogens Hepatovirus A – I and the avian Tremovirus pathogens, suggestive of a newly discovered pathogen of Northern Mockingbird. Although the recovered Hepeviridae virus is of unknown pathology, its family members include the Hepatitis E viruses. With the great diversity and novelty described from ileal viromes, discriminating potential pathogens and commensal microbiota from viruses associated with food items remains challenging. A deeper understanding of virus transmission and the risk of potential zoonosis can be enhanced by tracking viruses through the food web and via inter-specific and predator-prey interactions, particular in areas subject to land-use change, where human-wildlife interactions are increased and the risks from emerging pathogens of veterinary and medical importance are more pronounced.

Texas↗

Development and characterization of 12 polymorphic microsatellite loci in the sea sandwort, Honckenya peploides

Codominant marker systems are better suited to analyze population structure and assess the source of an individual in admixture analyses. Currently, there is no codominant marker system using microsatellites developed for the sea sandwort, Honckenya peploides (L.) Ehrh., an early colonizer in island systems. We developed and characterized novel microsatellite loci from H. peploides , using reads collected from whole genome shotgun sequencing on a 454 platform. The combined output from two shotgun runs yielded a total of 62,669 reads, from which 58 loci were screened. We identified 12 polymorphic loci that amplified reliably and exhibited disomic inheritance. Microsatellite data were collected and characterized for the 12 polymorphic loci in two Alaskan populations of H. peploides : Fossil Beach, Kodiak Island ( n = 32) and Egg Bay, Atka Island ( n = 29). The Atka population exhibited a slightly higher average number of alleles (3.9) and observed heterozygosity (0.483) than the Kodiak population (3.3 and 0.347, respectively). The overall probability of identity values for both populations was PID = 2.892e −6 and PID sib = 3.361e −3 . We also screened the 12 polymorphic loci in Wilhelmsia physodes (Fisch. ex Ser.) McNeill, the most closely related species to H. peploides , and only one locus was polymorphic. These microsatellite markers will allow future investigations into population genetic and colonization patterns of the beach dune ruderal H. peploides on new and recently disturbed islands.

Alaska↗

Are threatened seabird colonies of the Pacific Ocean genetically vulnerable? The case of the red-tailed tropicbird, Phaethon rubricauda, as a model species

Oceanic seabirds have suffered population declines and extirpations due to human disturbance and still face multiple threats. Here, we assessed the potential genetic vulnerability of the red-tailed tropicbird, Phaethon rubricauda , a seabird species threatened by human disturbance and listed as ‘least concern’ by the IUCN. Using Single Nucleotide Polymorphisms (SNPs) we evaluated the genetic population structure of the red-tailed tropicbird throughout the Pacific Ocean using samples from 132 individuals from six islands. We sampled individuals from islands without human-related disturbance (non-impacted islands) and with human-related disturbance (impacted islands). Results of genome-wide SNP analyses were consistent with previous results using mitochondrial DNA sequences analyses. Genetic diversity did not differ between impacted and non-impacted islands, and low inbreeding estimates were detected for all colonies. The SNPs analyses confirmed a pattern of isolation by distance and significant inter-regional (Chile, Australasia, and Hawaiʻi) genetic structure, but revealed greater differentiation of tropicbirds in Hawaiʻi compared with Chile and Australasia. Within regions, our results further indicated significant differentiation between Rapa Nui and Salas & Gómez Island (Chile), and between Meyer and Phillip islands (Australasia) that was not detected using mitochondrial DNA analyses. Within Hawaiʻi, we found a lack of significant genetic differentiation between Oʻahu and Kauaʻi, separated by 200 km. Our findings indicated that red-tailed tropicbird colonies are at genetic risk due to limited dispersal among colonies which may reduce the fitness of the species in the long-term. We suggest that red-tailed tropicbird colonies are vulnerable to future population declines because recovery through immigration from other islands may be limited by geographic distance. Conservation actions will help preserve genetic diversity and discrete populations for this native seabird at colonies throughout the Pacific.

Biodiversity and Conservation↗

Are captive tortoises a reservoir for conservation? An assessment of genealogical affiliation of captive Gopherus agassizii to local, wild populations

The conservation of tortoises poses a unique situation because several threatened species are commonly kept as pets within their native ranges. Thus, there is potential for captive populations to be a reservoir for repatriation efforts. We assess the utility of captive populations of the threatened Agassiz’s desert tortoise ( Gopherus agassizii ) for recovery efforts based on genetic affinity to local areas. We collected samples from 130 captive desert tortoises from three desert communities: two in California (Ridgecrest and Joshua Tree) and the Desert Tortoise Conservation Center (Las Vegas) in Nevada. We tested all samples for 25 short tandem repeats and sequenced 1,109 bp of the mitochondrial genome. We compared captive genotypes to a database of 1,258 Gopherus samples, including 657 wild caught G. agassizii spanning the full range of the species. We conducted population assignment tests to determine the genetic origins of the captive individuals. For our total sample set, only 44 % of captive individuals were assigned to local populations based on genetic units derived from the reference database. One individual from Joshua Tree, California, was identified as being a Morafka’s desert tortoise, G. morafkai , a cryptic species which is not native to the Mojave Desert. Our data suggest that captive desert tortoises kept within the native range of G. agassizii cannot be presumed to have a genealogical affiliation to wild tortoises in their geographic proximity. Precautions should be taken before considering the release of captive tortoises into the wild as a management tool for recovery.

Conservation Genetics↗

Implications of introgression for wildlife translocations: the case of North American martens

The evolutionary consequences of natural introgression provide a rare opportunity to retrospectively evaluate how the introduction of exotics or genetic rescue efforts may impact endemic faunas. Phylogeographic structure among mainland, endemic insular, and introduced North American marten ( Martes americana and M. caurina ) populations have been shaped by a complex history of natural, post-glacial population expansion followed by a series of anthropogenic introductions. In some cases, both natural colonization and translocations facilitated secondary contact, offering a series of replicated experiments that demonstrate how introgression, in these cases following isolation (insular and refugial), shapes genetic diversity. We test whether genetic exchange is occurring between North American marten species using mitochondrial genomes and ten nuclear loci. We present evidence of biased nuclear introgression from M. caurina into M. americana across two natural hybrid zones (insular and mainland) and found no remnant evidence of M. caurina on islands that received M. americana translocations, suggesting prior absence, potential extirpation, or genetic swamping of M. caurina from these islands. Our results highlight the importance of understanding phylogeographic variation prior to identifying source populations for wildlife translocations and caution the use of genetic rescue for North American marten populations. Although previously managed as a single species, these two species show substantial genetic divergence. When the two are placed into contact, they exhibit unidirectional, asymmetric introgression with potentially negative consequences for M. caurina , underscoring the value of mindful consideration of introgression in wildlife management.

Conservation Genetics↗

Genetic diversity and connectivity of moose (Alces americanus americanus) in eastern North America

Genetic diversity is critical to a population’s ability to overcome gradual environment change. Large-bodied wildlife existing in regions with relatively high human population density are vulnerable to isolation-induced genetic drift, population bottlenecks, and loss of genetic diversity. Moose ( Alces americanus americanus ) in eastern North America have a complex history of drastic population changes. Current and potential threats to moose populations in this region could be exacerbated by loss of genetic diversity and connectivity among subpopulations. Existing genetic diversity, gene flow, and population clustering and fragmentation of eastern North American moose are not well quantified, while physical and anthropogenic barriers to population connectivity already exist. Here, single nucleotide polymorphism (SNP) genotyping of 507 moose spanning five northeastern U.S. states and one southeastern Canadian province indicated low diversity, with a high proportion of the genomes sharing identity-by-state, with no consistent evidence of non-random mating. Gene flow estimates indicated bidirectionality between all pairs of sampled areas, with magnitudes reflecting clustering and differentiation patterns. A Discriminant Analysis of Principal Components analysis indicated that these genotypic data were best described with four clusters and indicated connectivity across the Saint Lawrence River and Seaway, a potential physical barrier to gene flow. Tests for genetic differentiation indicated restricted gene flow between populations across the Saint Lawrence River and Seaway, and between many sampled areas facing expanding human activity. These results document current genetic variation and connectivity of moose populations in eastern North America, highlight potential challenges to current population connectivity, and identify areas for future research and conservation.

New York, Vermont, New Hampshire, Maine, and Massa↗

A 37 K SNP array for the management and conservation of Golden Eagles (Aquila chrysaetos)

We describe the development of a custom 37 K Affymetrix Axiom myDesign single nucleotide polymorphism (SNP) array for a culturally and ecologically important apex predator, the golden eagle ( Aquila chrysaetos ). Using this SNP array, we performed population genomic analysis on 154 individuals of known natal localities and detected three genetic clusters that we designated as Taiga/High Arctic, Great Basin, and Rocky Mountains/Great Plains. Each of these clusters appears to display clinal variation within these geographic regions. After determining genetic structure, we performed an assignment test of 32 individuals, five of which were siblings of individuals used in the assessment of genetic structure, three had associated telemetry data, and the remaining individuals were of unknown natal locations. Using this array, four siblings were correctly assigned to the same geographic region as their sibling and the genetic assignment of the radio telemetered birds agreed with the expected movement patterns displayed by these individuals. For the remaining individuals, we were able to assign all but five individuals to one of the three genetic clusters. Our genetic assignments illustrates the utility of this SNP array to accurately assign most individuals to predesignated geographical regions. While further compiling genetic and other data types, we can increase the power of this tool for identifying those breeding populations that may need assistance due to anthropogenic stressors that negatively impact their population viability. The use of this genetic resource will help substantiate decisions by multiple conservation groups that seek to preserve the natural population structure of the golden eagle.

Conservation Genetics↗

Genetic Connectivity in the Arizona toad (Anaxyrus microscaphus): implications for conservation of a stream dwelling amphibian in the arid Southwestern U.S.

The Arizona Toad ( Anaxyrus microscaphus ) is restricted to riverine corridors and adjacent uplands in the arid southwestern United States. As with numerous amphibians worldwide, populations are declining and face various known or suspected threats, from disease to habitat modification resulting from climate change. The Arizona Toad has been petitioned to be listed under the U.S. Endangered Species Act and was considered “warranted but precluded” citing the need for additional information – particularly regarding natural history (e.g., connectivity and dispersal ability). The objectives of this study were to characterize population structure and genetic diversity across the species’ range. We used reduced-representation genomic sequencing to genotype 3,601 single nucleotide polymorphisms in 99 Arizona Toads from ten drainages across its range. Multiple analytical methods revealed two distinct genetic groups bisected by the Colorado River; one in the northwestern portion of the range in southwestern Utah and eastern Nevada and the other in the southeastern portion of the range in central and eastern Arizona and New Mexico. We also found subtle substructure within both groups, particularly in central Arizona where toads at lower elevations were less connected than those at higher elevations. The northern and southern parts of the Arizona Toad range are not well connected genetically and could be managed as separate units. Further, these data could be used to identify source populations for assisted migration or translocations to support small or potentially declining populations.

Arizona, Nevada, New Mexico↗

Population genetics of the endangered narrowly endemic Island Marble butterfly (Euchloe ausonides insulanus)

The Island Marble butterfly ( Euchloe ausonides insulanus ) is an endangered species endemic to the San Juan Islands off the coast of Washington State, United States, and British Columbia, Canada. The species was thought to be extinct for ~ 90 years before it was rediscovered at American Camp, San Juan Island National Historical Park in 1998. Here, we report the results of the first population genetic analyses for insulanus , using DNA collected non-invasively from individuals in the last known stronghold for the species. We used DNA extracted from meconium, larval exuviae, and natural mortalities to generate and test thirteen new microsatellite markers to estimate genetic diversity, population structure, and kinship. We assembled and annotated mitochondrial genomes, which were used alongside museum specimens of insulanus collected ~ 100 years ago from Vancouver Island, and other members of the E. ausonides species complex, to infer the evolutionary history of the species. The results indicated that insulanus experiences low heterozygosity, a small effective population size (N e ), and low allelic diversity. High levels of inbreeding were found in some individuals, but inbreeding was uneven across the population. No population structure or partitioning of genetic variation by host plant was detected. The mitogenomes of extant insulanus were all identical and modern samples showed a loss of allelic diversity compared to insulanus from museums. Extant insulanus formed a clade with museum specimens and we identified multiple putatively diagnostic alleles to differentiate insulanus from other subspecies. Based on these results, we outline considerations for species management and genetic monitoring.

British Columbia, Washington↗

Population connectivity and genetic structure of burbot ( Lota lota ) populations in the Wind River Basin, Wyoming

Burbot ( Lota lota ) occur in the Wind River Basin in central Wyoming, USA, at the southwestern extreme of the species’ native range in North America. The most stable and successful of these populations occur in six glacially carved mountain lakes on three different tributary streams and one large main stem impoundment (Boysen Reservoir) downstream from the tributary populations. Burbot are rarely found in connecting streams and rivers, which are relatively small and high gradient, with a variety of potential barriers to upstream movement of fish. We used high-throughput genomic sequence data for 11,197 SNPs to characterize the genetic diversity, population structure, and connectivity among burbot populations on the Wind River system. Fish from Boysen Reservoir and lower basin tributary populations were genetically differentiated from those in the upper basin tributary populations. In addition, fish within the same tributary streams fell within the same genetic clusters, suggesting there is movement of fish between lakes on the same tributaries but that populations within each tributary system are isolated and genetically distinct from other populations. Observed genetic differentiation corresponded to natural and anthropogenic barriers, highlighting the importance of barriers to fish population connectivity and gene flow in human-altered linked lake-stream habitats.

Wyoming↗

Development of a ddPCR assay for the detection of the Smoky Madtom (Noturus baileyi) from eDNA in stream water samples

The Smoky Madtom Noturus baileyi is a federally endangered species, whose native distribution includes lower Abrams Creek in Great Smoky Mountains National Park (GRSM) and Citico Creek in nearby Cherokee National Forest. Due to challenges for bio-monitoring posed by its nocturnality and cryptic life history, an environmental DNA (eDNA)-based approach for detection would be useful to complement existing electrofishing and seining efforts to better understand the distribution of this species. We developed a probe-based droplet digital PCR (ddPCR) assay to detect Smoky Madtoms from non-invasively collected water samples. The assay was specific to N. baileyi and did not amplify concentrated genomic DNA of 16 co-occurring or regional fish species, including the yellowfin madtom N. flavipinnis and stonecat N. flavus . The assay limit of detection (LOD) was determined to be 4.18 copies (95% CI: 3.95, 4.41). Several 2 L water samples collected from throughout various streams in GRSM in 2016 and 2017 were tested for the presence of N. baileyi using the ddPCR assay. N. baileyi was detected at two different sites in 2016 and 2017 within Abrams Creek previously known to contain N. baileyi , but no novel detections in other sampled streams were observed. This assay should prove useful for continued surveys of N. baileyi in GRSM.

Great Smoky Mountains National Park↗