Michael S. Eackles
Paul E Super
Matt A. Kulp
Becky J Nichols
Barbara A. Lubinski
Cheryl L. Morrison
Timothy L. King
Aaron W. Aunins
2022
<div id="Abs1-section" class="c-article-section"><div id="Abs1-content" class="c-article-section__content"><p>The Smoky Madtom<span> </span><i>Noturus baileyi</i><span> </span>is a federally endangered species, whose native distribution includes lower Abrams Creek in Great Smoky Mountains National Park (GRSM) and Citico Creek in nearby Cherokee National Forest. Due to challenges for bio-monitoring posed by its nocturnality and cryptic life history, an environmental DNA (eDNA)-based approach for detection would be useful to complement existing electrofishing and seining efforts to better understand the distribution of this species. We developed a probe-based droplet digital PCR (ddPCR) assay to detect Smoky Madtoms from non-invasively collected water samples. The assay was specific to<span> </span><i>N. baileyi</i><span> </span>and did not amplify concentrated genomic DNA of 16 co-occurring or regional fish species, including the yellowfin madtom<span> </span><i>N. flavipinnis</i><span> </span>and stonecat<span> </span><i>N. flavus</i>. The assay limit of detection (LOD) was determined to be 4.18 copies (95% CI: 3.95, 4.41). Several 2 L water samples collected from throughout various streams in GRSM in 2016 and 2017 were tested for the presence of<span> </span><i>N. baileyi</i><span> </span>using the ddPCR assay.<span> </span><i>N. baileyi</i><span> </span>was detected at two different sites in 2016 and 2017 within Abrams Creek previously known to contain<span> </span><i>N. baileyi</i>, but no novel detections in other sampled streams were observed. This assay should prove useful for continued surveys of<span> </span><i>N. baileyi</i><span> </span>in GRSM.</p></div></div>
application/pdf
10.1007/s12686-022-01290-3
en
Springer
Development of a ddPCR assay for the detection of the Smoky Madtom (Noturus baileyi) from eDNA in stream water samples
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