Folders: ROOT > ScienceBase Catalog > US Fish and Wildlife Service (USFWS) > Archive > Pacific Region, Region 1 > Pacific Lamprey Data Clearinghouse > Genetics > eDNA ( Show direct descendants )
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ROOT _ScienceBase Catalog __US Fish and Wildlife Service (USFWS) ___Archive ____Pacific Region, Region 1 _____Pacific Lamprey Data Clearinghouse ______Genetics _______eDNA Filters
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The Pacific lamprey (Entosphenus tridentatus) is an anadromous fish once abundant throughout coastal basins of western North America that has suffered dramatic declines in the last century due primarily to human activities. Here, we describe the development of an environmental DNA (eDNA) assay to detect Pacific lamprey in the Columbia River basin. The eDNA assay successfully amplified tissue derived DNA of Pacific lamprey collected from 12 locations throughout the Columbia River basin. The assay amplifies DNA from other Entosphenus species found outside of the Columbia River basin, but is species-specific within this basin. As a result, the assay presented here may be useful for detecting Entosphenus spp. in geographic...
This pilot study presents an environmental DNA (eDNA) assay for sea lamprey Petromyzon marinus and brown trout Salmo trutta, two species of economic and conservation importance in the Republic of Ireland. The results demonstrate the effectiveness of eDNA for assessing presence of low-abundance taxa (here, P. marinus) for environmental managers, and they highlight the potential for assessing relative abundance of rare or invasive freshwater species.
Genetic identification of aqueous environmental DNA (eDNA) provides site occupancy inferences for rare aquatic macrofauna that are often easier to obtain than direct observations of organisms. This relative ease makes eDNA sampling a valuable tool for conservation biology. Research on the origin, state, transport, and fate of eDNA shed by aquatic macrofauna is needed to describe the spatiotemporal context for eDNA-based occupancy inferences and to guide eDNA sampling design. We tested the hypothesis that eDNA is more concentrated in surficial sediments than in surface water by measuring the concentration of aqueous and sedimentary eDNA from an invasive fish, bigheaded Asian carp (Hypophthalmichthys spp.), in experimental...
Current research targeting filtered macrobial environmental DNA (eDNA) often relies upon cold ambient temperatures at various stages, including the transport of water samples from the field to the laboratory and the storage of water and/or filtered samples in the laboratory. This poses practical limitations for field collections in locations where refrigeration and frozen storage is difficult or where samples must be transported long distances for further processing and screening. This study demonstrates the successful preservation of eDNA at room temperature (20 °C) in two lysis buffers, CTAB and Longmire’s, over a 2-week period of time. Moreover, the preserved eDNA samples were seamlessly integrated into a phenol–chloroform–isoamyl...
Determining species distributions accurately is crucial to developing conservation and management strategies for imperiled species, but a challenging task for small populations. We evaluated the efficacy of environmental DNA (eDNA) analysis for improving detection and thus potentially refining the known distribution of Chinook salmon (Oncorhynchus tshawytscha) in the Methow and Okanogan Subbasins of the Upper Columbia River, which span the border between Washington, USA and British Columbia, Canada. We developed an assay to target a 90 base pair sequence of Chinook DNA and used quantitative polymerase chain reaction (qPCR) to quantify the amount of Chinook eDNA in triplicate 1-L water samples collected at 48 stream...
The discovery that macroorganisms can be detected from their environmental DNA (eDNA) in aquatic systems has immense potential for the conservation of biological diversity. This special issue contains 11 papers that review and advance the field of eDNA detection of vertebrates and other macroorganisms, including studies of eDNA production, transport, and degradation; sample collection and processing to maximize detection rates; and applications of eDNA for conservation using citizen scientists. This body of work is an important contribution to the ongoing efforts to take eDNA detection of macroorganisms from technical breakthrough to established, reliable method that can be used in survey, monitoring, and research...
Environmental DNA (eDNA) is DNA that has been released by an organism into its environment, such that the DNA can be found in air, water, or soil. In aquatic systems, eDNA has been shown to provide a sampling approach that is more sensitive for detecting target organisms faster, and less expensively than previous approaches. However, eDNA needs to be sampled in a manner that has been tested and found effective and, because single copies of target DNA are detected reliably, rigorous procedures must be designed to avoid sample contamination. Here we provide the details of a sampling protocol designed for detecting fish. This protocol, or very similar prototypes, has been used to collect data reported in multiple peerreviewed...
Three mantras often guide species and ecosystem management: (i) for preventing invasions by harmful species, ‘early detection and rapid response’; (ii) for conserving imperilled native species, ‘protection of biodiversity hotspots’; and (iii) for assessing biosecurity risk, ‘an ounce of prevention equals a pound of cure.’ However, these and other management goals are elusive when traditional sampling tools (e.g. netting, traps, electrofishing, visual surveys) have poor detection limits, are too slow or are not feasible. One visionary solution is to use an organism’s DNA in the environment (eDNA), rather than the organism itself, as the target of detection. In this issue of Molecular Ecology, Thomsen et al. (2012)...
Abstract Analysis of environmental DNA (eDNA) is an emerging technique used to detect aquatic species through water sampling and the extraction of biological material for amplification. Our study compared the efficacy of eDNA methodology to American Fisheries Society (AFS) standard snorkeling surveys with regard to detecting the presence of rare fish species. Knowing which method is more efficient at detecting target species will help managers to determine the best way to sample when both traditional sampling methods and eDNA sampling are available. Our study site included three Navajo Nation streams that contained Navajo Nation Genetic Subunit Bluehead Suckers Catostomus discobolus and Zuni Bluehead Suckers C....
EXECUTIVE SUMMARY: All living organisms shed DNA into the environment. Recent studies have demonstrated that environmental water samples can be effectively assayed for the DNA of target organisms. The DNA contained in these samples is commonly referred to as environmental DNA or eDNA. Because DNA assays can be sensitive to very small amounts of genetic material, eDNA surveys have the potential to detect sites where target organisms occur at very low abundances, potentially even when organisms are too rare for conventional survey methods to effectively detect them. We designed and tested twelve new eDNA markers for aquatic eDNA surveys of North American sturgeon. Eight of the markers are general for all North American...
Lampreys have a worldwide distribution, are functionally important to ecological communities and serve significant roles in many cultures. In Pacific coast drainages of North America, lamprey populations have suffered large declines. However, lamprey population status and trends within many areas of this region are unknown and such information is needed for advancing conservation goals. We developed two quantitative PCR-based, aquatic environmental DNA (eDNA) assays for detection of Pacific Lamprey (Entosphenus tridentatus) and Lampetra spp, using locked nucleic acids (LNAs) in the probe design. We used these assays to characterize the spatial distribution of lamprey in 18 watersheds of Puget Sound, Washington,...
Use of Droplet Digital PCR for Estimation of Fish Abundance and Biomass in Environmental DNA Surveys
An environmental DNA (eDNA) analysis method has been recently developed to estimate the distribution of aquatic animals by quantifying the number of target DNA copies with quantitative real-time PCR (qPCR). A new quantitative PCR technology, droplet digital PCR (ddPCR), partitions PCR reactions into thousands of droplets and detects the amplification in each droplet, thereby allowing direct quantification of target DNA. We evaluated the quantification accuracy of qPCR and ddPCR to estimate species abundance and biomass by using eDNA in mesocosm experiments involving different numbers of common carp.We found that ddPCR quantified the concentration of carp eDNA along with carp abundance and biomass more accurately...
Effective management of rare species, including endangered native species and recently introduced nonindigenous species, requires the detection of populations at low density. For endangered species, detecting the localized distribution makes it possible to identify and protect critical habitat to enhance survival or reproductive success. Similarly, early detection of an incipient invasion by a harmful species increases the feasibility of rapid responses to eradicate the species or contain its spread. Here we demonstrate the efficacy of environmental DNA (eDNA) as a detection tool in freshwater environments. Specifically, we delimit the invasion fronts of two species of Asian carps in Chicago, Illinois, USA area...
Abstract Environmental DNA (eDNA) is being rapidly adopted as a tool to detect rare animals. Quantitative PCR (qPCR) using probebased chemistries may represent a particularly powerful tool because of the method’s sensitivity, specificity, and potential to quantify target DNA. However, there has been little work understanding the performance of these assays in the presence of closely related, sympatric taxa. If related species cause any cross-amplification or interference, false positives and negatives may be generated. These errors can be disastrous if false positives lead to overestimate the abundance of an endangered species or if false negatives prevent detection of an invasive species. In this study we test...
The eDNAtlas is an open-access database developed through crowd-sourced field surveys that provides precise spatial information on the occurrence locations of aquatic species in the U.S. The eDNA samples constituting the database are collected using a standardized field sampling protocol by numerous natural resource agencies and non-governmental organizations partnered with the National Genomics Center for Wildlife and Fish Conservation (NGC), which is a science collaborative within the Rocky Mountain Research Station of the U.S. Forest Service. The eDNAtlas database contains results from thousands of sites and dozens of species and is annually updated with additional results for a growing list of species. Funding...
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