Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Mar 11;15(3):217.
doi: 10.3390/toxins15030217.

Artificial Substrates Coupled with qPCR (AS-qPCR) Assay for the Detection of the Toxic Benthopelagic Dinoflagellate Vulcanodinium rugosum

Aurélien Bouquet  1 , Christine Felix  1 , Estelle Masseret  2 , Coralie Reymond  2 , Eric Abadie  1   3 , Mohamed Laabir  2 , Jean Luc Rolland  1
Affiliations

Artificial Substrates Coupled with qPCR (AS-qPCR) Assay for the Detection of the Toxic Benthopelagic Dinoflagellate Vulcanodinium rugosum

Aurélien Bouquet et al. Toxins (Basel). .

Abstract

Vulcanodinium rugosum is an emerging benthopelagic neuro-toxic dinoflagellate species responsible for seasonal Pinnatoxins and Portimines contaminations of shellfish and marine animals. This species is challenging to detect in the environment, as it is present in low abundance and difficult to be identified using light microscopy. In this work, we developed a method using artificial substrates coupled with qPCR (AS-qPCR) to detect V. rugosum in a marine environment. This sensitive, specific and easy-to-standardize alternative to current techniques does not require specialized expertise in taxonomy. After determining the limits and specificity of the qPCR, we searched for the presence of V. rugosum in four French Mediterranean lagoons using artificial substrates collected every two weeks for one year. The AS-qPCR method revealed its occurrences in summer 2021 in every studied lagoon and detected cells in more samples than light microscopy. As V. rugosum development induces shellfish contamination even at low microalga densities, the AS-qPCR method is accurate and relevant for monitoring V. rugosum in a marine environment.

Keywords: PCR; Vulcanodinium rugosum; artificial substrate; benthopelagic; detection; toxins.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Amplification results of qPCR products from 6-fold diluted Vulcanodinium rugosum DNA extract: (A) Amplification curves plot, (B) standard curve, and (C) derivative melting curves plot. The error bars represent ± standard deviation from triplicate mean values. MT: melting temperature.
Figure 2
Figure 2
Collecting stations in Thau, Ingril, Vic and Prévost.
Figure 3
Figure 3
Phylogenetic distance tree of Vulcanodinium rugosum LSRU genes used to design PCR primers.
Figure 4
Figure 4
Structure of Vulcanodinium rugosum species rDNA and location of primers for the V. rugosum genus-specific PCR. NTS: non-transcribed spacer; SSU: small subunit; LSU: large subunit.
See this image and copyright information in PMC

References

    1. Moreira A.R., Comas A., Valle A., Seisdedo M. Bloom of Vulcanodinium rugosum Linked to Skin Lesions in Cienfuegos Bay, Cuba. IOC Newsl. Toxic Algae Algal Bloom. 2016;55:10–11. doi: 10.1016/j.scitotenv.2020.143782. - DOI
    1. Rhodes L., Smith K., Selwood A., McNabb P., Munday R., Suda S., Molenaar S., Hallegraeff G. Dinoflagellate Vulcanodinium rugosum Identified as the Causative Organism of Pinnatoxins in Australia, New Zealand and Japan. Phycologia. 2011;50:624–628. doi: 10.2216/11-19.1. - DOI - PubMed
    1. Smith K.F., Rhodes L.L., Suda S., Selwood A.I. A Dinoflagellate Producer of Pinnatoxin G, Isolated from Sub-Tropical Japanese Waters. Harmful Algae. 2011;10:702–705. doi: 10.1016/j.hal.2011.05.006. - DOI
    1. Zeng N., Gu H., Smith K.F., Rhodes L.L., Selwood A.I., Yang W. The First Report of Vulcanodinium rugosum (Dinophyceae) from the South China Sea with a Focus on the Life Cycle. N. Z. J. Mar. Freshw. Res. 2012;46:511–521. doi: 10.1080/00288330.2012.719911. - DOI
    1. Hess P., Abadie E., Herve F., Berteaux T., Sechet V., Araoz R., Molgo J., Zakarian A., Sibat M., Rundberget T., et al. Is Responsible for Atypical Toxicity in Mussels (Mytilus galloprovincialis) and Clams (Venerupis decussata) from Ingril, a French Mediterranean Lagoon. Toxicon Off. J. Int. Soc. Toxinology. 2013;75:16–26. doi: 10.1016/j.toxicon.2013.05.001. - DOI - PMC - PubMed

Publication types