A Noninvasive Tool to Assess the Distribution of Pacific Lamprey (Entosphenus tridentatus) in the Columbia River Basin.

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 range beyond the Columbia River Basin. The assay did not amplify tissue or synthetically derived DNA of 14 commonly sympatric non-target species, including lampreys of the genus Lampetra, which are morphologically similar to Pacific lamprey in the freshwater larval stage

Primer sequences for amplification and sequencing of the COI region of the Pacific lamprey mitochondrial genome; and sequences for components of the eDNA assay.

<p>Primer sequences for amplification and sequencing of the COI region of the Pacific lamprey mitochondrial genome; and sequences for components of the eDNA assay.</p

Collection and species assemblage information for eDNA samples used to test the Pacific lamprey qPCR assay.

<p>Community assemblage information (fish species present) was obtained via a combination of eDNA analysis and personal communication with local tribal, state and federal biologists.</p

List of species used for <i>in vitro</i> screening of the qPCR primers and probe.

<p>Origin refers to the waterbody for Pacific lamprey samples; origin is listed as state for all other samples.</p

Species and source for DNA sequences used for <i>in silico</i> assay development.

<p>Source refers to GenBank accession number or location of collection for specimens analyzed in this study. Probe mismatches refers to the number of base-pair differences between the eDNA assay probe and the COI sequence for a particular species. Pacific brook lamprey (<i>Lampetra pacifica</i>) sequence data were obtained after primer and probe development.</p