Evaluation of DNA metabarcoding for identifying fish eggs: a case study on the West Florida Shelf

A critical factor in fisheries management is the protection of spawning sites for ecologically and economically important fish species. DNA barcoding (i.e., amplification and sequencing of the mitochondrial cytochrome c oxidase I (COI) gene) of fish eggs has emerged as a powerful technique for ident...

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Main Authors: Mya Breitbart, Makenzie Kerr, Michael J. Schram, Ian Williams, Grace Koziol, Ernst Peebles, Christopher D. Stallings
Format: Article
Language:English
Published: PeerJ Inc. 2023-03-01
Series:PeerJ
Subjects:
Online Access:https://peerj.com/articles/15016.pdf
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author Mya Breitbart
Makenzie Kerr
Michael J. Schram
Ian Williams
Grace Koziol
Ernst Peebles
Christopher D. Stallings
author_facet Mya Breitbart
Makenzie Kerr
Michael J. Schram
Ian Williams
Grace Koziol
Ernst Peebles
Christopher D. Stallings
author_sort Mya Breitbart
collection DOAJ
description A critical factor in fisheries management is the protection of spawning sites for ecologically and economically important fish species. DNA barcoding (i.e., amplification and sequencing of the mitochondrial cytochrome c oxidase I (COI) gene) of fish eggs has emerged as a powerful technique for identifying spawning sites. However, DNA barcoding of individual fish eggs is time-consuming and expensive. In an attempt to reduce costs and effort for long-term fisheries monitoring programs, here we used DNA metabarcoding, in which DNA is extracted and amplified from a composited sample containing all the fish eggs collected at a given site, to identify fish eggs from 49 stations on the West Florida Shelf. A total of 37 taxa were recovered from 4,719 fish eggs. Egg distributions on the West Florida Shelf corresponded with the known habitat types occupied by these taxa, which included burrower, coastal pelagic, epipelagic, mesopelagic, demersal, deep demersal, commensal, and reef-associated taxa. Metabarcoding of fish eggs was faster and far less expensive than barcoding individual eggs; however, this method cannot provide absolute taxon proportions due to variable copy numbers of mitochondrial DNA in different taxa, different numbers of cells within eggs depending on developmental stage, and PCR amplification biases. In addition, some samples yielded sequences from more taxa than the number of eggs present, demonstrating the presence of contaminating DNA and requiring the application of a threshold proportion of sequences required for counting a taxon as present. Finally, we review the advantages and disadvantages of using metabarcoding vs. individual fish egg barcoding for long-term monitoring programs.
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spelling doaj.art-e930db9bc95843808a788d995cd7b8f52023-12-03T10:23:08ZengPeerJ Inc.PeerJ2167-83592023-03-0111e1501610.7717/peerj.15016Evaluation of DNA metabarcoding for identifying fish eggs: a case study on the West Florida ShelfMya BreitbartMakenzie KerrMichael J. SchramIan WilliamsGrace KoziolErnst PeeblesChristopher D. StallingsA critical factor in fisheries management is the protection of spawning sites for ecologically and economically important fish species. DNA barcoding (i.e., amplification and sequencing of the mitochondrial cytochrome c oxidase I (COI) gene) of fish eggs has emerged as a powerful technique for identifying spawning sites. However, DNA barcoding of individual fish eggs is time-consuming and expensive. In an attempt to reduce costs and effort for long-term fisheries monitoring programs, here we used DNA metabarcoding, in which DNA is extracted and amplified from a composited sample containing all the fish eggs collected at a given site, to identify fish eggs from 49 stations on the West Florida Shelf. A total of 37 taxa were recovered from 4,719 fish eggs. Egg distributions on the West Florida Shelf corresponded with the known habitat types occupied by these taxa, which included burrower, coastal pelagic, epipelagic, mesopelagic, demersal, deep demersal, commensal, and reef-associated taxa. Metabarcoding of fish eggs was faster and far less expensive than barcoding individual eggs; however, this method cannot provide absolute taxon proportions due to variable copy numbers of mitochondrial DNA in different taxa, different numbers of cells within eggs depending on developmental stage, and PCR amplification biases. In addition, some samples yielded sequences from more taxa than the number of eggs present, demonstrating the presence of contaminating DNA and requiring the application of a threshold proportion of sequences required for counting a taxon as present. Finally, we review the advantages and disadvantages of using metabarcoding vs. individual fish egg barcoding for long-term monitoring programs.https://peerj.com/articles/15016.pdfFishEggBarcodingMetabarcodingFloridaCOI
spellingShingle Mya Breitbart
Makenzie Kerr
Michael J. Schram
Ian Williams
Grace Koziol
Ernst Peebles
Christopher D. Stallings
Evaluation of DNA metabarcoding for identifying fish eggs: a case study on the West Florida Shelf
PeerJ
Fish
Egg
Barcoding
Metabarcoding
Florida
COI
title Evaluation of DNA metabarcoding for identifying fish eggs: a case study on the West Florida Shelf
title_full Evaluation of DNA metabarcoding for identifying fish eggs: a case study on the West Florida Shelf
title_fullStr Evaluation of DNA metabarcoding for identifying fish eggs: a case study on the West Florida Shelf
title_full_unstemmed Evaluation of DNA metabarcoding for identifying fish eggs: a case study on the West Florida Shelf
title_short Evaluation of DNA metabarcoding for identifying fish eggs: a case study on the West Florida Shelf
title_sort evaluation of dna metabarcoding for identifying fish eggs a case study on the west florida shelf
topic Fish
Egg
Barcoding
Metabarcoding
Florida
COI
url https://peerj.com/articles/15016.pdf
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