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At least 379 records · Page 21Linked to original sources
USGS2018

Deciphering the link between doubly uniparental inheritance of mtDNA and sex determination in bivalves: Clues from comparative transcriptomics

Bivalves exhibit an astonishing diversity of sexual systems and sex-determining mechanisms. They can be gonochoric, hermaphroditic or androgenetic, with both genetic and environmental factors known to determine or influence sex. One unique sex-determining system involving the mitochondrial genome has also been hypothesized to exist in bivalves with doubly uniparental inheritance (DUI) of mtDNA. However, the link between DUI and sex determination remains obscure. In this study, we performed a comparative gonad transcriptomics analysis for two DUI-possessing freshwater mussel species to better understand the mechanisms underlying sex determination and DUI in these bivalves. We used a BLAST reciprocal analysis to identify orthologs between Venustaconcha ellipsiformis and Utterbackia peninsularis and compared our results with previously published sex-specific bivalve transcriptomes to identify conserved sex-determining genes. We also compared our data with other DUI species to identify candidate genes possibly involved in the regulation of DUI. A total of ∼12,000 orthologous relationships were found, with 2,583 genes differentially expressed in both species. Among these genes, key sex-determining factors previously reported in vertebrates and in bivalves (e.g., Sry, Dmrt1, Foxl2 ) were identified, suggesting that some steps of the sex-determination pathway may be deeply conserved in metazoans. Our results also support the hypothesis that a modified ubiquitination mechanism could be responsible for the retention of the paternal mtDNA in male bivalves, and revealed that DNA methylation could also be involved in the regulation of DUI. Globally, our results suggest that sets of genes associated with sex determination and DUI are similar in distantly-related DUI species.

Genome Biology and Evolution↗
USGS2024

Metagenomic sequencing sheds light on microbes putatively associated with pneumonia-related fatalities of white-tailed deer (Odocoileus virginianus)

With emerging infectious disease outbreaks in human, domestic and wild animal populations on the rise, improvements in pathogen characterization and surveillance are paramount for the protection of human and animal health, as well as the conservation of ecologically and economically important wildlife. Genomics offers a range of suitable tools to meet these goals, with metagenomic sequencing facilitating the characterization of whole microbial communities associated with emerging and endemic disease outbreaks. Here, we use metagenomic sequencing in a case-control study to identify microbes in lung tissue associated with newly observed pneumonia-related fatalities in 34 white-tailed deer ( Odocoileus virginianus ) in Wisconsin, USA. We identified 20 bacterial species that occurred in more than a single individual. Of these, only Clostridium novyi was found to substantially differ (in number of detections) between case and control sample groups; however, this difference was not statistically significant. We also detected several bacterial species associated with pneumonia and/or other diseases in ruminants ( Mycoplasma ovipneumoniae , Trueperella pyogenes , Pasteurella multocida , Anaplasma phagocytophilum , Fusobacterium necrophorum ); however, these species did not substantially differ between case and control sample groups. On average, we detected a larger number of bacterial species in case samples than controls, supporting the potential role of polymicrobial infections in this system. Importantly, we did not detect DNA of viruses or fungi, suggesting that they are not significantly associated with pneumonia in this system. Together, these results highlight the utility of metagenomic sequencing for identifying disease-associated microbes. This preliminary list of microbes will help inform future research on pneumonia-associated fatalities of white-tailed deer.

Microbial Genomics↗
USGS2016

H9N2 influenza A virus isolated from a Greater White-fronted wild goose (Anser albifrons) in Alaska has a mutation in the PB2 gene, which is associated with pathogenicity in human pandemic 2009 H1N1

We report here the genomic sequence of an H9N2 influenza A virus [A/greater white-fronted goose/Alaska/81081/2008 (H9N2)]. This virus shares ≥99.8% identity with a previously reported virus. Both strains contain a G590S mutation in the polymerase basic 2 (PB2) gene, which is a pathogenicity marker in the pandemic 2009 H1N1 virus when combined with R591.

Genome Announcements↗
USGS2015

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

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

Virginia↗
USGS2020

Subspecies differentiation in an enigmatic chaparral shrub species

Premise Delimiting biodiversity units is difficult in organisms in which differentiation is obscured by hybridization, plasticity, and other factors that blur phenotypic boundaries. Such work is more complicated when the focal units are subspecies, the definition of which has not been broadly explored in the era of modern genetic methods. Eastwood manzanita ( Arctostaphylos glandulosa Eastw.) is a widely distributed and morphologically complex chaparral shrub species with much subspecific variation, which has proven challenging to categorize. Currently 10 subspecies are recognized, however, many of them are not geographically segregated, and morphological intermediates are common. Subspecies delimitation is of particular importance in this species because two of the subspecies are rare. The goal of this study was to apply an evolutionary definition of “subspecies” to characterize structure within Eastwood manzanita. Methods We used publicly available geospatial environmental data and reduced‐representation genome sequencing to characterize environmental and genetic differentiation among subspecies. In addition, we tested whether subspecies could be differentiated by environmentally associated genetic variation. Results Our analyses do not show genetic differentiation among subspecies of Eastwood manzanita, with the exception of one of the two rare subspecies. In addition, our environmental analyses did not show ecological differentiation, though limitations of the analysis prevent strong conclusions. Conclusions Genetic structure within Eastwood manzanita does not correspond to current subspecies circumscriptions, but rather reflects geographic distribution. Our study suggests that subspecies concepts need to be reconsidered in long‐lived plant species, especially in the age of next‐generation sequencing.

American Journal of Botany↗
USGS2016

Assessing models of speciation under different biogeographic scenarios; An empirical study using multi-locus and RNA-seq analyses

Evolutionary biology often seeks to decipher the drivers of speciation, and much debate persists over the relative importance of isolation and gene flow in the formation of new species. Genetic studies of closely related species can assess if gene flow was present during speciation, because signatures of past introgression often persist in the genome. We test hypotheses on which mechanisms of speciation drove diversity among three distinct lineages of desert tortoise in the genus Gopherus . These lineages offer a powerful system to study speciation, because different biogeographic patterns (physical vs. ecological segregation) are observed at opposing ends of their distributions. We use 82 samples collected from 38 sites, representing the entire species' distribution and generate sequence data for mtDNA and four nuclear loci. A multilocus phylogenetic analysis in *BEAST estimates the species tree. RNA‐seq data yield 20,126 synonymous variants from 7665 contigs from two individuals of each of the three lineages. Analyses of these data using the demographic inference package ∂a∂i serve to test the null hypothesis of no gene flow during divergence. The best‐fit demographic model for the three taxa is concordant with the *BEAST species tree, and the ∂a∂i analysis does not indicate gene flow among any of the three lineages during their divergence. These analyses suggest that divergence among the lineages occurred in the absence of gene flow and in this scenario the genetic signature of ecological isolation (parapatric model) cannot be differentiated from geographic isolation (allopatric model).

Ecology and Evolution↗
USGS2018

A new tool for studying waterfowl immune and metabolic responses: Molecular level analysis using kinome profiling

Here, we describe the design of an Anas‐ specific kinome peptide array that can be used to study the immunometabolic responses of mallard and American black duck to pathogens, contaminants, and environmental stress. The peptide arrays contain 2,642 unique phosphorylate‐able peptide sequences representing 1,900 proteins. These proteins cover a wide array of metabolic and immunological processes, and 758 Gene Ontology Biological processes are statistically significantly represented on the duck peptide array of those 164 contain the term “metabolic” and 25 “immune.” In addition, we conducted a comparison of mallard to American black duck at a genetic and proteomic level. Our results show a significant genomic and proteomic overlap between these two duck species, so that we have designed a cross‐reactive peptide array capable of studying both species. This is the first reported development of a wildlife species‐specific kinome peptide array.

Ecology and Evolution↗
USGS2019

North-facing slopes and elevation shape asymmetric genetic structure in the range-restricted salamander Plethodon shenandoah

Species with narrow environmental preferences are often distributed across fragmented patches of suitable habitat, and dispersal among subpopulations can be difficult to directly observe. Genetic data collected at population centers can help quantify gene flow, which is especially important for vulnerable species with a disjunct range. Plethodon shenandoah is a Federally Endangered salamander known only from three mountaintops in Virginia, USA. To reconstruct the evolutionary history and population connectivity of this species, we generated both mitochondrial and nuclear data using sequence capture for all three populations and found strong population structure that was independent of geographic distance. Both the nuclear markers and mitochondrial genome indicated a deep split between the most southern population and the combined central and northern population. Although there was some mitochondrial haplotype-splitting between the central and northern populations, there was complete admixture in nuclear markers. This is indicative of either a recent split or current male-biased dispersal among mountain isolates. Models of landscape resistance found that dispersal across north-facing slopes at mid-elevation levels best explain the observed genetic structure among populations. These unexpected results highlight the importance of landscape features in understanding and predicting movement and fragmentation of salamanders across space.

Ecology and Evolution↗
USGS2016

Estimating heterotrophic respiration at large scales: Challenges, approaches, and next steps

Heterotrophic respiration (HR), the aerobic and anaerobic processes mineralizing organic matter, is a key carbon flux but one impossible to measure at scales significantly larger than small experimental plots. This impedes our ability to understand carbon and nutrient cycles, benchmark models, or reliably upscale point measurements. Given that a new generation of highly mechanistic, genomic-specific global models is not imminent, we suggest that a useful step to improve this situation would be the development of “Decomposition Functional Types” (DFTs). Analogous to plant functional types (PFTs), DFTs would abstract and capture important differences in HR metabolism and flux dynamics, allowing modelers and experimentalists to efficiently group and vary these characteristics across space and time. We argue that DFTs should be initially informed by top-down expert opinion, but ultimately developed using bottom-up, data-driven analyses, and provide specific examples of potential dependent and independent variables that could be used. We present an example clustering analysis to show how annual HR can be broken into distinct groups associated with global variability in biotic and abiotic factors, and demonstrate that these groups are distinct from (but complementary to) already-existing PFTs. A similar analysis incorporating observational data could form the basis for future DFTs. Finally, we suggest next steps and critical priorities: collection and synthesis of existing data; more in-depth analyses combining open data with rigorous testing of analytical results; using point measurements and realistic forcing variables to constrain process-based models; and planning by the global modeling community for decoupling decomposition from fixed site data. These are all critical steps to build a foundation for DFTs in global models, thus providing the ecological and climate change communities with robust, scalable estimates of HR.

Ecosphere↗
USGS2025

Evaluation of rapid DNA extraction methods to better enable point-of-use environmental DNA detection

Recent developments in molecular testing have created the opportunity for biologists and managers to detect environmental DNA (eDNA) of target species rapidly and without the requirement of a laboratory. These point-of-use protocols may be especially useful for early detection and rapid response for invasive species or surveillance for at-risk native species, where timely management decisions are critical. Point-of-use eDNA protocols also facilitate wider and less expensive implementation of eDNA methods. One of the key components to an effective point-of-use protocol is a rapid DNA extraction method. Several rapid extraction protocols are suitable for implementation in the field, but information regarding their relative effectiveness is lacking. We evaluated extraction efficiency of four DNA rapid extraction protocols using filters spiked with primary cultured grass carp ( Ctenopharyngodon idella ) gill cells. The extraction methods included two syringe-based column extractions, a lysis and extraction solution, and a divalent cation chelation resin (Chelex) extraction protocol alongside a laboratory-based control kit. We estimated DNA yield using a newly designed quantitative polymerase chain reaction (qPCR) assay targeting the grass carp nuclear genome. We evaluated two additional factors, filter type (mixed cellulose ester [MCE] and polyethersulfone [PES]) and background eDNA source (aquaculture or river). The lysis and extraction solution and Chelex extraction both had the highest overall yield, with MCE filters further increasing Chelex yield while the enzyme extraction yield was dependent on interaction with both filter and eDNA source. Our results indicate that rapid extraction protocols, such as solutions with short heating steps, are effective for DNA isolation and help to increase the overall accessibility of eDNA analyses.

Environmental DNA↗
USGS2022

Do pharmaceuticals in the environment pose a risk to wildlife?

The vast majority of knowledge related to the question of, “To what extent do pharmaceuticals in the environment pose a risk to wildlife?”, stems from the Asian vulture crisis (>99% decline of some species of old-world vultures on the Indian subcontinent related to the veterinary use of the non-steroidal anti-inflammatory drug (NSAID) diclofenac). The hazard of diclofenac and other NSAIDs (carprofen, flunixin, ketoprofen, nimesulide, phenylbutazone) to vultures and other avian species has since been demonstrated; indeed only meloxicam and tolfenamic acid have been found to be vulture-safe. Since diclofenac was approved for veterinary use in Spain and Italy in 2013 (home to ~95% of vultures in Europe), the risk of NSAIDs to vultures in these countries has become one of the principal concerns related to pharmaceuticals and wildlife. Many of the other bodies of work on pharmaceutical exposure, hazard and risk to wildlife also relate to adverse effects in birds, (e.g., poisoning of scavenging birds in North America and Europe from animal carcasses containing pentobarbital; secondary and even tertiary poisoning of birds exposed to pesticides used in veterinary medicine as cattle dips; migratory birds as a vector for the transfer of antimicrobial and antifungal resistance). While there is some research related to endocrine disruption in reptiles and potential exposure of aerial insectivores, there remain numerous knowledge gaps for risk posed by pharmaceuticals to amphibians, reptiles and mammals. Developing non-invasive sampling techniques and new approach methodologies (e.g., genomic, in vitro , in silico , in ovo ) are important if we are to bridge the current knowledge gaps without extensive vertebrate testing.

Environmental Toxicology and Chemistry↗
USGS2014

Beta-thymosin gene polymorphism associated with freshwater invasiveness of alewife ( Alosa pseudoharengus )

Predicting the success of a species’ colonization into a novel environment is routinely considered to be predicated on niche-space similarity and vacancy, as well as propagule pressure. The role genomic variation plays in colonization success (and the interaction with environment) may be suggested, but has not rigorously been documented. To test an hypothesis that previously observed ecotype-specific polymorphisms between anadromous and landlocked alewife (Alosa pseudoharengus) populations are an adaptive response to osmoregulatory challenges rather than a result of allele sampling at founding, we examined multiple anadromous and landlocked (colonized) populations for their allelic profiles at a conserved region (3’-UTR end) of a β-thymosin gene whose protein product plays a central role in the organization of cytoskeleton. The putatively ancestral β-thymosin allele was prevalent in anadromous populations, whereas a newly derived allele was overrepresented in landlocked populations; a third allele was exclusive to the anadromous populations. We also conducted a complementary set of salinity exposure experiments to test osmoregulatory performance of the alewife ecotypes in contrasting saline environments. The pattern of variation and results from these challenges indicate a strong association of β-thymosin with colonization success and a transition for species with an anadromous life-history to one with only a freshwater component.

Journal of Experimental Zoology↗
USGS2014

A tetrapod-like repertoire of innate immune receptors and effectors for coelacanths

The recent availability of both robust transcriptome and genome resources for coelacanth ( Latimeria chalumnae ) has led to unique discoveries for coelacanth immunity such as the lack of IgM, a central component of adaptive immunity. This study was designed to more precisely address the origins and evolution of gene families involved in the initial recognition and response to microbial pathogens, which effect innate immunity. Several multigene families involved in innate immunity are addressed, including: Toll-like receptors (TLRs), retinoic acid inducible gene 1 (RIG1)-like receptors (RLRs), the nucleotide-binding domain and leucine-rich repeat containing proteins (NLRs), diverse immunoglobulin domain-containing proteins (DICP) and modular domain immune-type receptors (MDIRs). Our analyses also include the tripartite motif-containing proteins (TRIM), which are involved in pathogen recognition as well as the positive regulation of antiviral immunity. Finally, this study addressed some of the downstream effectors of the antimicrobial response including IL-1 family members, type I and II interferons (IFN) and IFN-stimulated effectors (ISGs). Collectively, the genes and gene families in coelacanth that effect innate immune functions share characteristics both in content, structure and arrangement with those found in tetrapods but not in teleosts. The findings support the sister group relationship of coelacanth fish with tetrapods.

Journal of Experimental Zoology Part B: Molecular ↗
USGS2021

Stock composition of the historical New York Bight Atlantic sturgeon (Acipenser oxyrinchus oxyrinchus) intercept fishery revealed through microsatellite analysis of archived spines

A targeted commercial fishery for Atlantic Sturgeon Acipenser oxyrinchus oxyrinchus once operated in the New York Bight, where it was assumed that most harvested Atlantic Sturgeon were natal to the Hudson River population. However, more recent evidence suggests that the fishery may have been targeting a mixed-stock aggregation, in which case harvested Atlantic Sturgeon could have been comprised of individuals from multiple populations throughout the species’ range. Although there is now a moratorium on Atlantic Sturgeon harvest in the New York Bight, modern molecular approaches provide an opportunity to use archived tissues to perform a retrospective mixed-stock analysis on the fishery. Genomic DNA extracted from archived fin spines from 80 Atlantic Sturgeon collected nearly 30 years ago suggests that the fishery primarily harvested individuals from the Hudson River population. However, based on individual-based assignment tests, our results indicate that the fishery also harvested individuals from at least eight other populations located throughout the species’ range. This study highlights how archival hard parts that were previously used for age and growth analyses can be employed for retrospective genetic analyses. Further, because the New York Bight harbors relatively high concentrations of Atlantic Sturgeon, the study shows how localized management decisions can influence Atlantic Sturgeon conservation at rangewide scales. When integrated with more recent knowledge of species ecology, these analyses can be used to evaluate the efficacy of previous management strategies and understand the effects of historical processes on contemporary demography.

New York↗
USGS2018

Hybridization and genetic structure of Neosho Smallmouth Bass in the Ozark Highlands

The Neosho Smallmouth Bass Micropterus dolomieu velox is endemic to Arkansas River tributaries originating in the Ozark Highlands and Boston Mountains. Although morphologically and genetically distinct from other populations of Smallmouth Bass M. dolomieu , the conservation‐genetic status of Neosho Smallmouth Bass is largely unknown. To assist in filling this data gap, we quantified introgressive hybridization, genetic diversity, and population structure of Neosho Smallmouth Bass in two major river basins (Grand River and Illinois River) using seven polymorphic microsatellite markers. Introgressive hybridization with stocked Tennessee lake‐strain Smallmouth Bass was most prevalent in the Illinois River, wherein the overall genomic proportion of Neosho Smallmouth Bass alleles was only 0.422. After accounting for hybrid individuals, genetic diversity of Neosho Smallmouth Bass was generally higher in larger rivers and lower in smaller, isolated streams. Three distinct population clusters were identified at the uppermost level of genetic structure—one from the Illinois River basin and two from the Grand River basin. These three population boundaries accounted for approximately 7% of the hierarchical genetic variation within our data set, and substructure below the uppermost level accounted for an additional 2% of genetic variation. The population structure we discovered can provide a blueprint for management that conserves diversity within and among populations; for example, population boundaries can be used to determine brood source locations for supplemental stocking efforts to counter nonnative introgression. Introgressive hybridization with nonnative Smallmouth Bass appears to be the most pertinent threat to the Neosho Smallmouth Bass subspecies, which is of conservation value to the overall adaptability of Smallmouth Bass as a species.

Missouri, Oklahoma↗
USGS2022

Population connectivity of aquatic insects in a dam-regulated, desert river

Humans have exaggerated natural habitat fragmentation, negatively impacting species dispersal and reducing population connectivity. Habitat fragmentation can be especially detrimental in freshwater populations, whose dispersal is already constrained by the river network structure. Aquatic insects, for instance, are generally limited to two primary modes of dispersal: downstream drift in the aquatic juvenile life stages and flight during the terrestrial winged adult stage. Yet the impacts of large hydropower dams can make rivers uninhabitable for incoming (drifting) juvenile insects, with remaining refugia found only in tributaries. The ability of adult aquatic insects to traverse such river stretches in search of suitable tributary habitat likely depends on factors such as species-specific dispersal ability and distance between tributaries. To explore the intersection of natural and human-induced habitat fragmentation on aquatic insect dispersal ability, we quantified population genetics of three taxa with varying dispersal abilities, a caddisfly (Hydropsychidae, Hydropsyche oslari ), a mayfly (Baetidae: Fallceon quilleri ), and a water strider (Veliidae: Rhagovelia distincta ), throughout tributaries of the Colorado River in the Grand Canyon, Arizona, USA. Using 2bRAD reduced genome sequencing and landscape genetics analyses, we revealed a strong pattern of isolation by distance among mayfly populations. This contrasts with caddisfly and water strider populations, which were largely panmictic. Analysis of thousands of informative single nucleotide polymorphisms showed that realized dispersal ability may not be accurately predicted by species traits for these widespread species. Principal components analysis revealed a strong division between caddisfly populations upstream and downstream of Havasu Creek (279 km through the 390 km study reach), suggesting that the geography of the Grand Canyon imposes a dispersal barrier for this species. Our use of genetic tools in the Grand Canyon to understand population structure has enabled us to elucidate dispersal barriers for aquatic insects. Ultimately, these data may be useful in informing effective conservation management plans for understudied organisms of conservation interest.

River Research and Applications↗
USGS2018

Combining genetic, isotopic, and field data to better describe the influence of dams and diversions on Burbot Movement in the Wind River Drainage, Wyoming

Dams and water diversions fragment habitat, entrain fish, and alter fish movement. Many Burbot Lota lota populations are declining, with dams and water diversions thought to be a major threat. We used multiple methods to identify Burbot movement patterns and assess entrainment into an irrigation system in the Wind River, Wyoming. We assessed seasonal movement of Burbot with a mark–recapture (PIT tagging) study, natal origins of entrained fish with otolith microchemistry, and historic movement with genotyping by sequencing. We found limited evidence of entrainment in irrigation waters across all approaches. The mark–recapture study indicated that out‐migration from potential source populations could be influenced by flow regime but was generally low. Otolith and genomic results suggested the presence of a self‐sustaining population within the irrigation network. We conclude that emigration from natural tributary populations is not the current source of the majority of Burbot found in irrigation waters. Instead, reservoir and irrigation canal construction has created novel habitat in which Burbot have established a population. Using a multi‐scale approach increased our inferential abilities and mechanistic understanding of movement patterns between natural and managed systems.

Wyoming↗
USGS2019

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

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

Transactions of the American Fisheries Society↗