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. 2017 Sep 5;12(9):e0183347.
doi: 10.1371/journal.pone.0183347. eCollection 2017.

DNA in a bottle-Rapid metabarcoding survey for early alerts of invasive species in ports

Yaisel J Borrell  1 , Laura Miralles  1 , Hoang Do Huu  2 , Khaled Mohammed-Geba  3 , Eva Garcia-Vazquez  1
Affiliations

DNA in a bottle-Rapid metabarcoding survey for early alerts of invasive species in ports

Yaisel J Borrell et al. PLoS One. .

Abstract

Biota monitoring in ports is increasingly needed for biosecurity reasons and safeguarding marine biodiversity from biological invasion. Present and future international biosecurity directives can be accomplished only if the biota acquired by maritime traffic in ports is controlled. Methodologies for biota inventory are diverse and now rely principally on extensive and labor-intensive sampling along with taxonomic identification by experts. In this study, we employed an extremely simplified environmental DNA (eDNA) sampling methodology from only three 1-L bottles of water per port, followed by metabarcoding (high-throughput sequencing and DNA-based species identification) using 18S rDNA and Cytochrome oxidase I as genetic barcodes. Eight Bay of Biscay ports with available inventory of fouling invertebrates were employed as a case study. Despite minimal sampling efforts, three invasive invertebrates were detected: the barnacle Austrominius modestus, the tubeworm Ficopomatus enigmaticus and the polychaete Polydora triglanda. The same species have been previously found from visual and DNA barcoding (genetic identification of individuals) surveys in the same ports. The current costs of visual surveys, conventional DNA barcoding and this simplified metabarcoding protocol were compared. The results encourage the use of metabarcoding for early biosecurity alerts.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. Map showing the eight ports analysed in this study. International cargo ports are marked with black squares and fishing ports and marinas with white squares.
Numbers from 1 to 8 are Figueras (Eo), Luarca, Cudillero, Aviles, Gijon, Villaviciosa, Ribadesella and Luarca.
Fig 2
Fig 2. Non-metric Multidimensional Scaling of the metabarcodes found for 18S rDNA and Cytochrome oxidase I gene in the analysed ports. A: Shepard plot; B: Scatter plot.
The port names are given and genes acronyms are 18S for 18S rDNA and COI for Cytochrome oxidase I gene. The 95% ellipsis for the data is shown in scatter plot.
See this image and copyright information in PMC

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