Impact Factor 4.019

The world's most-cited Microbiology journal

Methods ARTICLE Provisionally accepted The full-text will be published soon. Notify me

Front. Microbiol. | doi: 10.3389/fmicb.2018.03153

qPCR assays for the detection and quantification of multiple Paralytic Shellfish Toxin-producing species of Alexandrium

 Rendy Ruvindy1*,  Christopher J. Bolch2, Lincoln MacKenzie3, Kirsty Smith3 and  Shauna A. Murray1*
  • 1Climate Change Cluster, University of Technology Sydney, Australia
  • 2Institute for Marine and Antarctic Studies (IMAS), Australia
  • 3Cawthron Institute, New Zealand

Paralytic shellfish toxin producing dinoflagellates have negatively impacted the shellfish aquaculture industry worldwide, including in Australia and New Zealand. Morphologically identical cryptic species of dinoflagellates that may differ in toxicity, in particular, species of the former Alexandrium tamarense species complex, co-occur in Australia, as they do in multiple regions in Asia and Europe. To understand the dynamics and the ecological drivers of the growth of each species in the field, accurate quantification at the species level is crucial. We have developed the first quantitative polymerase chain reaction (qPCR) primers for A. australiense, and new primers targeting A. ostenfeldii, A. catenella, and A. pacificum. We showed that our new primers for A. pacificum are more specific than previously published primer pairs. These assays can be used to quantify planktonic cells and cysts in the water column and in sediment samples with limits of detection of 2 cells/L for the A. catenella and A. australiense assays, 2 cells/L and 1 cyst/mg sediment for the A. pacificum assay, and 1 cells/L for the A. ostenfeldii assay, and efficiencies of > 90%. We utilised these assays to discriminate and quantify co-occurring A. catenella, A. pacificum and A. australiense in samples from the east coast of Tasmania, Australia.

Keywords: Paralytic Shellfish toxin (PST), Alexandrium, qPCR, ribosomal DNA, dinoflagellate cysts

Received: 23 Aug 2018; Accepted: 05 Dec 2018.

Edited by:

Senjie Lin, University of Connecticut, United States

Reviewed by:

Deana Erdner, University of Texas at Austin, United States
BRENDAN P. BURNS, University of New South Wales, Australia
Samuel Cirés, Universidad Autónoma de Madrid, Spain  

Copyright: © 2018 Ruvindy, Bolch, MacKenzie, Smith and Murray. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence:
Mr. Rendy Ruvindy, Climate Change Cluster, University of Technology Sydney, Sydney, 2007, Australia, r.ruvindy@gmail.com
Dr. Shauna A. Murray, Climate Change Cluster, University of Technology Sydney, Sydney, 2007, Australia, shauna.murray@uts.edu.au