Effectiveness of DNA barcoding for identifying pis

Authors: Z Moran D J Orth J D Schmitt EM Hallerman R Aguilar

Publish Date: 2015/09/07

Volume: 99, Issue: 1, Pages: 161-167

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Abstract

Introduced predators pose ecological impacts upon prey species and receiving ecosystems Understanding such ecological interactions creates technical challenges including speciesspecific identification of partially digested prey items in the stomachs of piscivorous predators We present the first evaluation of DNA barcoding to identify piscine prey in the stomachs of North American catfishes Family Ictaluridae Fish prey items of nonnative Blue Catfish Ictalurus furcatus and Flathead Catfish Pylodictis olivaris were obtained by gastric lavage and ranked as lightly moderately or heavily digested We used an established cocktail of universal fish primers FishF2 t1 FishR2 t1 VF2 t1 and FR1d t1 to amplify the cytochrome oxidase I COI3 region of mitochondrial DNA from these samples Amplification products were subjected to Sanger sequencing and edited sequences were compared to entries in GenBank Eightysix percent of the sequences generated for lightly or moderately digested samples and 66  of those for heavily digested samples could be assigned to the species level based on similarity with archived COI3 sequences While traditional morphological identification led to specieslevel identification of 65  of fish prey items addition of DNA barcoding resulted in identification to species of 88  of fish prey items overall Diet items identified by DNA markers included anadromous Striped Bass Morone saxatilis and herrings and shads Alosa spp that are the focus of fishery restoration programs in these rivers We found DNA barcoding to be an efficient and costeffective addition to diet studies of nonnative predatorsThis study was completed with funds provided by the Virginia Department of Game and Inland Fisheries through a Sport Fish Restoration Grant from the US Fish and Wildlife Service Funding for the participation of EMH and DJO was provided in part by the Virginia Agricultural Experiment Station and the Hatch Program of the National Institute of Food and Agriculture US Department of Agriculture All applicable international national and/or institutional guidelines for the care and use of animals were followed Special thanks to Jason Emmel and Tim Lane for their assistance with field collections and laboratory analysis This report was strengthened by attention to the comments of two anonymous peer reviewers

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