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ABSTRACT The Indo‐Pacific lionfish,Pterois volitans,is an invasive species in the western Atlantic. Since its introduction to Florida in the early 1980s, populations have surged with lionfish now found from North Carolina to Venezuela. As their range expands, these generalist predators threaten native fauna, and while they are primarily a marine species, their tolerance for low salinity conditions may allow them to expand into sensitive estuarine habitats undetected. Traditional approaches for tracking invasive species such as direct observation or trapping are impractical over large spatial scales, making environmental DNA (eDNA) an attractive alternative. Molecular assays, such as those employing quantitative polymerase chain reaction (qPCR), amplify low copy number DNA fragments in environmental samples and are increasingly employed as a complement to traditional methods for the detection of invasive species. Currently, there is one published PCR assay for the detection of lionfish eDNA. However, the specificity of this assay is unverified, and the critical performance parameters limit of detection (LOD) and limit of quantification (LOQ) have not been established. Here we evaluate the efficacy of this assay and show that it is likely to result in false negatives in the western Atlantic. As an alternative, we developed a new TaqMan probe‐based qPCR assay that is species‐specific forP. volitansand highly sensitive with a LOD of 12 copies per reaction and a LOQ of 598 copies per reaction. While our assay does not amplify the closely relatedP. miles, which was also introduced in the western Atlantic, the low prevalence of this species in the invasive population means our assay is effective for most monitoring purposes. We conclude that our assay is a robust method for the detection of lionfish and can be employed in any habitat, offering new opportunities for controlling the spread of invasive lionfish.
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Hidden in plain sight: The invasive macroalga Caulerpa prolifera evades detection by environmental DNA methods
Abstract Environmental managers need a rapid and cost‐effective monitoring tool for tracking the spread of invasive species, particularly at the onset of introduction. The macroalgaeCaulerpa proliferais considered an invasive species outside its native range, colonizing large patches of seafloor, reducing native species, and altering ecosystem functioning. Here, we developed a droplet digital PCR assay for detection ofC. proliferafrom environmental DNA seawater samples using the internal transcribed spacer (ITS) region. While the assay itself was confirmed to be highly efficient, we discovered concentrations ofC. proliferaeDNA were present below detectable levels in the water column surrounding an outbreak. To understand why, we conducted tank‐based experiments for two California invasive algae species,Caulerpa proliferaandSargassum horneri. The steady‐state eDNA concentration (eDNA copies/ gram of biomass detected) ofC. proliferawas found to be two orders of magnitude lower thanS. horneri. A meta‐analysis of steady‐state concentrations reported in the literature showed a remarkable range from ~104–1011(copies/g), revealing C. proliferato have the lowest recorded steady‐state concentrations of eDNA of any known species. We attributeC. prolifera'slow steady‐state eDNA concentration to its unique biology as a unicellular macroscopic algae which reduces the possible modes of eDNA release compared to similarly sized multicellular organisms. Critically our results demonstrate the potential limits of eDNA approaches, the influence of shedding rates in the reliability of species detections, and the vital importance of benchmarking and validating eDNA assays in both field and laboratory settings.
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- Award ID(s):
- 2024426
- PAR ID:
- 10493465
- Publisher / Repository:
- Wiley
- Date Published:
- Journal Name:
- Environmental DNA
- Volume:
- 6
- Issue:
- 1
- ISSN:
- 2637-4943
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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Abstract BackgroundEnvironmental DNA (eDNA) is an effective tool for the detection and monitoring of presence or absence of rare and invasive species. These techniques have been extended to quantify biomass in vertebrates, particularly in fish species. However, the efficacy of eDNA techniques to quantify biomass in invertebrate species has rarely been examined. This study tested whether eDNA could be used to determine the biomass of the world-wide invasive green crab,Carcinus maenas. In a controlled laboratory study, the relationship between biomass andC. maenaseDNA concentration was examined in the context of different biotic (activity) and abiotic (temperature) parameters. ResultsWhen incubating different numbers of crabs in sterile saltwater for up to 7 days, a relationship between eDNA concentration and biomass was observed at temperatures of 6.7 ℃ and 18.7 ℃, but not at 12.8 ℃. Additionally, motor activity, aggression level, time of sampling, and features of organismal decay had significant impact on the concentration ofC. maenaseDNA collected. ConclusionsWe show that eDNA concentration did not correlate with biomass, and that biomass, temperature, organismal characteristics, and potentially many more parameters affect shedding and degradation rates for eDNA in this species, thus, impacting the recoverable eDNA concentration. Therefore, eDNA techniques are not likely to provide a reliable signal of biomass in the invasive invertebrate speciesC. maenas.more » « less
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Abstract The early detection of invasive species is essential to cease the spread of the species before it can cause irreversible damage to the environment. The analysis of environmental DNA (eDNA) has emerged as a non-harmful method to detect the presence of a species before visual detection and is a promising approach to monitor invasive species. Few studies have investigated the use of eDNA for arthropods, as their exoskeleton is expected to limit the release of eDNA into the environment. We tested published primers for the invasive European green crab,Carcinus maenas, in the Gulf of Maine and found them not species-specific enough for reliable use outside of the area for which they were designed for. We then designed new primers, tested them against a broad range of local faunal species, and validated these primers in a field study. We demonstrate that eDNA analyses can be used for crustaceans with an exoskeleton and suggest that primers and probe sequences must be tested on local fauna at each location of use to ensure no positive amplification of these other species.more » « less
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- Free Publicly Accessible Full Text
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Accepted Manuscript1.0
- Journal Article:
- https://doi.org/10.1002/edn3.496
