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Use of DNA barcoding to detect invertebrate invasive species from diapausing eggs

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Abstract

The global transhipment of ballast water and associated flora and fauna by cargo vessels has increased dramatically in recent decades. Invertebrate species are frequently carried in ballast water and sediment, although identification of diapausing eggs can be extremely problematic. Here we test the application of DNA barcoding using mitochondrial cytochrome c oxidase subunit I and 16S rDNA to identify species from diapausing eggs collected in ballast sediment of ships. The accuracy of DNA barcoding identification was tested by comparing results from the molecular markers against each other, and by comparing barcoding results to traditional morphological identification of individuals hatched from diapausing eggs. Further, we explored two public genetic databases to determine the broader applicability of DNA barcodes. Of 289 diapausing eggs surveyed, sufficient DNA for barcoding was obtained from 96 individuals (33%). Unsuccessful DNA extractions from 67% of eggs in our study were most likely due to degraded condition of eggs. Of 96 eggs with successful DNA extraction, 61 (64%) were identified to species level, while 36% were identified to possible family/order level. Species level identifications were always consistent between methodologies. DNA barcoding was suitable for a wide range of taxa, including Branchiopoda, Copepoda, Rotifera, Bryozoa and Ascidia. Branchiopoda and Copepoda were respectively the best and worst represented groups in genetic databases. Though genetic databases remain incomplete, DNA barcoding resolved nearly double the number of species identified by traditional taxonomy (19 vs. 10). Notorious invaders are well represented in existing databases, rendering these NIS detectable using molecular methods. DNA barcoding provides a rapid and accurate approach to identification of invertebrate diapausing eggs that otherwise would be very difficult to identify.

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Acknowledgments

We thank the Shipping Federation of Canada and the multiple shipping companies that facilitated access to vessels, and our ballast sampling teams: C. van Overdijk, A.M. Weise, O. Casas-Monroy, N. Simard, J.-Y. Couture, M. Huot and Dr. C. McKindsey. We are grateful for taxonomic assistance from Drs. S.I. Dodson, J.R. Cordell and P. Hudson, and laboratory support from R. Tedla, S. Ross and H. Coker. Great thanks to Drs. Aibin Zhan and Francisco Sylvester for commenting on an early version of the manuscript. Comments from two anonymous reviewers are gratefully acknowledged. This research was supported by NSERC’s Canadian Aquatic Invasive Species Network, Transport Canada, Fisheries and Oceans Canada, by NSERC Discovery Grants to MEC, SAB and HJM, and by a DFO Invasive Species Research Chair to HJM.

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Authors and Affiliations

  1. Great Lakes Institute for Environmental Research, University of Windsor, Windsor, Ontario, N9B 3P4, Canada

    Elizabeta Briski, Melania E. Cristescu, Sarah A. Bailey & Hugh J. MacIsaac

  2. Great Lakes Laboratory for Fisheries and Aquatic Sciences, Fisheries and Oceans Canada, 867 Lakeshore Road, Burlington, Ontario, L7R 4A6, Canada

    Sarah A. Bailey

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  1. Elizabeta Briski
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  2. Melania E. Cristescu
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  3. Sarah A. Bailey
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  4. Hugh J. MacIsaac
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Correspondence to Elizabeta Briski.

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Table 4 List of species identified by morphological methods and DNA barcodes
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Briski, E., Cristescu, M.E., Bailey, S.A. et al. Use of DNA barcoding to detect invertebrate invasive species from diapausing eggs. Biol Invasions 13, 1325–1340 (2011). https://doi.org/10.1007/s10530-010-9892-7

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