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Development of a real-time polymerase chain reaction assay for the detection of the invasive Mediterranean fanworm, Sabella spallanzanii, in environmental samples

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Abstract

The Mediterranean fanworm, Sabella spallanzanii Gmelin 1791, was first detected in the Southern Hemisphere in the 1990s and is now abundant in many parts of southern Australia and in several locations around northern New Zealand. Once established, it can proliferate rapidly, reaching high densities with potential ecological and economic impacts. Early detection of new S. spallanzanii incursions is important to prevent its spread, guide eradication or control efforts and to increase knowledge on the species’ dispersal pathways. In this study, we developed a TaqMan probe real-time polymerase chain reaction assay targeting a region of the mitochondrial cytochrome oxidase I gene. The assay was validated in silico and in vitro using DNA from New Zealand and Australian Sabellidae with no cross-reactivity detected. The assay has a linear range of detection over seven orders of magnitude with a limit of detection reached at 12.4 × 10−4 ng/μL of DNA. We analysed 145 environmental (water, sediment and biofouling) samples and obtained positive detections only from spiked samples and those collected at a port where S. spallanzanii is known to be established. This assay has the potential to enhance current morphological and molecular-based methods, through its ability to rapidly and accurately identify S. spallanzanii in environmental samples.

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Acknowledgements

This work was funded by NIWA under Coasts and Oceans Research Programme 6, Marine Biosecurity (2015/16 SCI). We thank Barrie Forrest and Grant Hopkins (Cawthron) and Chris Woods (NIWA) for native Sabellidae collection, Fiona Gower (Cawthron) for taxonomic identification of specimens, the National Invertebrates Collection (NIWA, Wellington) for providing S. spallanzanii and native Sabellidae specimens, Janet Adamson and Witold Ming (Cawthron) for technical assistance, and Nathan Bott (RMIT University, Australia) for providing DNA from the Australian polychaetes. We thank Oliver Floerl (Cawthron) for helpful comments on the draft manuscript and Ulla von Ammon and Olivier Laroche (Cawthron) for supplying their unpublished HTS data and DNA extracts for analysis in this study.

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Authors and Affiliations

  1. Coastal and Freshwater Group, Cawthron Institute, Nelson, New Zealand

    Susanna A. Wood, Anastasija Zaiko, Ingrid Richter & Xavier Pochon

  2. Environmental Research Institute, University of Waikato, Hamilton, New Zealand

    Susanna A. Wood

  3. Marine Science and Technology Centre, Klaipeda University, Klaipeda, Lithuania

    Anastasija Zaiko

  4. National Institute of Water and Atmospheric Research Ltd, Christchurch, New Zealand

    Graeme J. Inglis

  5. Institute of Marine Science, University of Auckland, Auckland, New Zealand

    Xavier Pochon

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  1. Susanna A. Wood
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  2. Anastasija Zaiko
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  3. Ingrid Richter
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  4. Graeme J. Inglis
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  5. Xavier Pochon
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Correspondence to Susanna A. Wood.

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Responsible editor: Philippe Garrigues

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Wood, S.A., Zaiko, A., Richter, I. et al. Development of a real-time polymerase chain reaction assay for the detection of the invasive Mediterranean fanworm, Sabella spallanzanii, in environmental samples. Environ Sci Pollut Res 24, 17373–17382 (2017). https://doi.org/10.1007/s11356-017-9357-y

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