Using eDNA sampling for species-specific fish detection in tropical oceanic samples: limitations and recommendations for future use

Background Over the past decade, environmental DNA (eDNA) has become a resourceful tool in conservation and biomonitoring. Environmental DNA has been applied in a variety of environments, but the application to studies of marine fish, particularly at tropical latitudes, are limited. Since many comme...

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Main Authors: Giovanna M. Gonzalez Colmenares, Alejandro J. Gonzalez Montes, Chelsea A. Harms-Tuohy, Nikolaos V. Schizas
Format: Article
Language:English
Published: PeerJ Inc. 2023-02-01
Series:PeerJ
Subjects:
Online Access:https://peerj.com/articles/14810.pdf
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author Giovanna M. Gonzalez Colmenares
Alejandro J. Gonzalez Montes
Chelsea A. Harms-Tuohy
Nikolaos V. Schizas
author_facet Giovanna M. Gonzalez Colmenares
Alejandro J. Gonzalez Montes
Chelsea A. Harms-Tuohy
Nikolaos V. Schizas
author_sort Giovanna M. Gonzalez Colmenares
collection DOAJ
description Background Over the past decade, environmental DNA (eDNA) has become a resourceful tool in conservation and biomonitoring. Environmental DNA has been applied in a variety of environments, but the application to studies of marine fish, particularly at tropical latitudes, are limited. Since many commercially important Caribbean fishes are overexploited, these species are optimal candidates to explore the use of this method as a biomonitoring tool. Specifically, for many of these species, the formation of fish spawning aggregations (FSAs) marks a critical life history event where fishes will gather in large numbers for reproduction. These FSAs are ephemeral in nature, lasting only a few days, but are predictable in time and space which makes them susceptible to overfishing. Methods In this study, we test the feasibility of using an eDNA sampling approach (water and sediment collection) to detect the presence of known FSAs off the west coast of Puerto Rico, with cytochrome c oxidase subunit 1 (CO1) and 12S rRNA (12S) primers designed to target specific species. A total of 290 eDNA samples were collected and, of those, 206 eDNA samples were processed. All eDNA samples varied in DNA concentration, both between replicates and collection methods. A total of 12 primer sets were developed and tested using traditional PCR and qPCR. Results Despite validation of primer accuracy and sample collection during known peak spawning times, the use of traditional PCR and qPCR with both molecular markers failed to produce species-specific amplification. Thus, a trial test was conducted using the CO1 primers in which target fish DNA was ‘spiked’ at various concentrations into the respective eDNA samples to determine the target species DNA concentration limit of detection. Upon successful amplification of the trial, results indicated that eDNA samples were below the detection threshold of our methods, suggesting that the number of fish present at the spawning aggregations was inadequate for single-species detection methods. In addition, elements such as the unavoidable presence of non-target DNA, oceanic environmental conditions, shedding rates of target fish, among other biotic and abiotic factors could have affected DNA persistence and degradation rates at the sites. Conclusion We provide recommendations for species-specific fish detection in lower latitudes, and suggestions for studies aiming to monitor or detect fish spawning aggregations using eDNA sampling.
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spelling doaj.art-111526d650e546d1aca35c113ac9e0a22023-12-03T12:46:17ZengPeerJ Inc.PeerJ2167-83592023-02-0111e1481010.7717/peerj.14810Using eDNA sampling for species-specific fish detection in tropical oceanic samples: limitations and recommendations for future useGiovanna M. Gonzalez Colmenares0Alejandro J. Gonzalez Montes1Chelsea A. Harms-Tuohy2Nikolaos V. Schizas3Department of Biology, Universidad de Puerto Rico, Recinto de Mayagüez, Mayagüez, Puerto RicoDepartment of Marine Sciences, Universidad de Puerto Rico, Recinto de Mayagüez, Mayagüez, Puerto RicoIsla Mar Research Expeditions, Rincón, Puerto RicoDepartment of Marine Sciences, Universidad de Puerto Rico, Recinto de Mayagüez, Mayagüez, Puerto RicoBackground Over the past decade, environmental DNA (eDNA) has become a resourceful tool in conservation and biomonitoring. Environmental DNA has been applied in a variety of environments, but the application to studies of marine fish, particularly at tropical latitudes, are limited. Since many commercially important Caribbean fishes are overexploited, these species are optimal candidates to explore the use of this method as a biomonitoring tool. Specifically, for many of these species, the formation of fish spawning aggregations (FSAs) marks a critical life history event where fishes will gather in large numbers for reproduction. These FSAs are ephemeral in nature, lasting only a few days, but are predictable in time and space which makes them susceptible to overfishing. Methods In this study, we test the feasibility of using an eDNA sampling approach (water and sediment collection) to detect the presence of known FSAs off the west coast of Puerto Rico, with cytochrome c oxidase subunit 1 (CO1) and 12S rRNA (12S) primers designed to target specific species. A total of 290 eDNA samples were collected and, of those, 206 eDNA samples were processed. All eDNA samples varied in DNA concentration, both between replicates and collection methods. A total of 12 primer sets were developed and tested using traditional PCR and qPCR. Results Despite validation of primer accuracy and sample collection during known peak spawning times, the use of traditional PCR and qPCR with both molecular markers failed to produce species-specific amplification. Thus, a trial test was conducted using the CO1 primers in which target fish DNA was ‘spiked’ at various concentrations into the respective eDNA samples to determine the target species DNA concentration limit of detection. Upon successful amplification of the trial, results indicated that eDNA samples were below the detection threshold of our methods, suggesting that the number of fish present at the spawning aggregations was inadequate for single-species detection methods. In addition, elements such as the unavoidable presence of non-target DNA, oceanic environmental conditions, shedding rates of target fish, among other biotic and abiotic factors could have affected DNA persistence and degradation rates at the sites. Conclusion We provide recommendations for species-specific fish detection in lower latitudes, and suggestions for studies aiming to monitor or detect fish spawning aggregations using eDNA sampling.https://peerj.com/articles/14810.pdfeDNA samplingSpecies-specific detectionFish spawning aggregationsTropical eDNAFisheries management
spellingShingle Giovanna M. Gonzalez Colmenares
Alejandro J. Gonzalez Montes
Chelsea A. Harms-Tuohy
Nikolaos V. Schizas
Using eDNA sampling for species-specific fish detection in tropical oceanic samples: limitations and recommendations for future use
PeerJ
eDNA sampling
Species-specific detection
Fish spawning aggregations
Tropical eDNA
Fisheries management
title Using eDNA sampling for species-specific fish detection in tropical oceanic samples: limitations and recommendations for future use
title_full Using eDNA sampling for species-specific fish detection in tropical oceanic samples: limitations and recommendations for future use
title_fullStr Using eDNA sampling for species-specific fish detection in tropical oceanic samples: limitations and recommendations for future use
title_full_unstemmed Using eDNA sampling for species-specific fish detection in tropical oceanic samples: limitations and recommendations for future use
title_short Using eDNA sampling for species-specific fish detection in tropical oceanic samples: limitations and recommendations for future use
title_sort using edna sampling for species specific fish detection in tropical oceanic samples limitations and recommendations for future use
topic eDNA sampling
Species-specific detection
Fish spawning aggregations
Tropical eDNA
Fisheries management
url https://peerj.com/articles/14810.pdf
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