Open AccessDevelopment and validation of an environmental DNA protocol to detect an invasive Caribbean freshwater fish, the guppy ( Poecilia reticulata )
Author(s) -
Smith Sierra N.,
Schlupp Ingo,
Higgins Edward D.,
Watters Jessa L.,
Bennett KerriAnn,
Bräger Stefan,
Siler Cameron D.
Publication year - 2022
Publication title -
environmental dna
Language(s) - English
Resource type - Journals
ISSN - 2637-4943
DOI - 10.1002/edn3.248
Subject(s) - poecilia , guppy , biology , environmental dna , freshwater fish , zoology , mitochondrial dna , cytochrome b , ecology , fish <actinopterygii> , fishery , biodiversity , gene , genetics
We describe the development and validation of a qPCR assay to detect Poecilia reticulata , a highly invasive species of freshwater fish invasive to the Caribbean islands, through environmental DNA (eDNA) sampling. Originating from Trinidad, this species is invasive and detrimental to countless native tropical fish communities. A qPCR assay, consisting of a set of primers and a fluorescent probe, amplifying a 214 base pair target region of the mitochondrial Cytochrome B gene was designed for P . reticulata from existing DNA sequence data. The assay was assessed for target specificity, with no evidence of amplification in closely related or sympatrically distributed non‐target species. In vitro tests indicate that the assay consistently detects P . reticulata down to concentrations of 2.0 × 10 −5 ng/μl. The developed assay provides a new, practical tool for monitoring freshwater habitats throughout the Caribbean, allowing for early and rapid detection of invasive fish species of conservation concern.