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The update and optimization of an eDNA assay to detect the invasive rusty crayfish (Faxonius rusticus)
- Source :
- PLoS ONE, PLoS ONE, Vol 16, Iss 10, p e0259084 (2021), PLoS ONE, Vol 16, Iss 10 (2021)
- Publication Year :
- 2021
- Publisher :
- Public Library of Science (PLoS), 2021.
-
Abstract
- Environmental DNA (eDNA) is nuclear or mitochondrial DNA shed into the environment, and amplifying this DNA can serve as a reliable, noninvasive way to monitor aquatic systems for the presence of an invasive species. Assays based on the collection of eDNA are becoming increasingly popular, and, when optimized, can aid in effectively and efficiently tracking invasion fronts. We set out to update an eDNA assay to detect the invasive rusty crayfish, Faxonius rusticus. We tested for species specificity compared to other stream crayfish and field tested the assay at sites with known presence (N = 3) and absence (N = 4) in the Juniata River watershed in central Pennsylvania, USA. To maximize sensitivity, we field tested different storage buffers (Longmire’s buffer and ethanol), DNA extraction methods (Qiagen’s DNEasy and PowerWater kits), and quantitative polymerase chain reaction (qPCR) chemistries (TaqMan and SYBR green). Our assay confirmed the presence data and performed optimally when filter samples were stored in Longmire’s buffer, DNA was extracted with DNeasy Blood and Tissue Kit, and TaqMan qPCR chemistry was utilized. With proper sample processing, our assay allows for accurate, noninvasive detection of F. rusticus in streams.
- Subjects :
- Physiology
Invasive Species
Artificial Gene Amplification and Extension
Astacoidea
Polymerase Chain Reaction
law.invention
Specimen Storage
law
Medicine and Health Sciences
Environmental DNA
Signal Amplification
DNA extraction
Polymerase chain reaction
Multidisciplinary
Organic Compounds
Eukaryota
Crayfish
Crustaceans
Body Fluids
Chemistry
Blood
Real-time polymerase chain reaction
Physical Sciences
Medicine
Engineering and Technology
Anatomy
Research Article
Environmental Monitoring
Mitochondrial DNA
Arthropoda
Science
Biology
Extraction techniques
Species Colonization
Rivers
TaqMan
Animals
Molecular Biology Techniques
Molecular Biology
Chromatography
Ethanol
Organic Chemistry
Ecology and Environmental Sciences
Organisms
Chemical Compounds
Biology and Life Sciences
Rusty crayfish
Pennsylvania
biology.organism_classification
Invertebrates
DNA, Environmental
Research and analysis methods
Alcohols
Storage and Handling
Signal Processing
Introduced Species
Zoology
Subjects
Details
- ISSN :
- 19326203
- Volume :
- 16
- Database :
- OpenAIRE
- Journal :
- PLOS ONE
- Accession number :
- edsair.doi.dedup.....618f7b8ea08ff753be6cd8eca63b55a1