1. Our team boasts extensive experience in the development, management and coordination of large scientific research projects. We use highly rigorous experimental designs and novel molecular techniques to examine phycotoxins and their causative species in relation to the Australian shellfish industry. Our project objectives focus mainly on Tasmania, but also include results that are relevant to the wide Australian shellfish industry and are as follows:OBJECTIVE 1. GENERATE KNOWLEDGE ABOUT COMMERCIAL DST TEST KITS AND RAPID MOLECULAR TECHNIQUES (SUCH AS qPCR) FOR DST TOXIN AND SPECIES DETECTION• Conduct a desktop literature review to determine the most up-to-date information on available commercial test kits, the toxin analogs that they target and their capabilities. • Conduct literature review to determine the most up-to-date information on rapid molecular techniques for the detection of DST producing dinoflagellates belonging to the genus Dinophysis.
2. OBJECTIVE 2. IDENTIFY DST PROFILES PRESENT IN AUSTRALIAN SHELLFISH AND ASSESS LABORATORY CAPABILITIES TO DETECT THESE TOXINS• Define and synthesise information on the precise DST toxin analogue/profiles in Australian shellfish (oysters, mussels and pipis) in relation to the Dinophysis species that are currently present in Australian coastal waters (Dinophysis acuminata, D. caudata, D. fortii, D. tripos, D. acuta, D. ovum, D. truncata, Prorocentrum spp.)• Conduct an initial assessment of analytical laboratories in the detection of DSTs in shellfish (oysters, mussels and pipis). A full spike and recovery quality control assessment program to assess the analytical techniques and to determine whether the type of sample matrix affects toxin detection will be undertaken. Shellfish matrices (oysters, mussels and pipis) will be spiked with known quantities of toxin standards in varying quantities and sent to 4 analytical laboratories (Cawthron Institute, Sydney Institute of Marine Science, Analytical Services Tasmania, and Symbio Laboratories) for LC-MS/MS toxin separation and confirmation.• Assess and conduct statistical analysis on results.• Work with NSW Food Authority and other state regulators/farmers to obtain DSTs positive shellfish from Australia-wide shellfish from farmers (when a toxic episode occurs) and analyse toxin profiles from event sampling. • Report findings to laboratory managers, relevant authorities and all other industry partners/collaborators.
3. OBJECTIVE 3. COMPARE THE EFFICACY OF COMMERCIALLY AVAILABLE TOXIN DETECTING KITS USING RELEVANT SAMPLE MATRICES • Conduct a replicated, quality-controlled laboratory-based study to compare the efficacy of a range of commercially available rapid test kits with DSTs standards spiked in oyster, mussel and pipi samples. Toxin kits to be tested will include Zeulab/Abraxis, Beacon, BioScientific, Biosens/Abraxis, Europroxima, Neogen and Scotia, and any new commercially available kits found during the literature review.
4. OBJECTIVE 4. DEVELOP A QUANTITATIVE PCR ASSAY FOR DINOPHYSIS SPECIES DETECTION FOR POTENTIAL ONSITE FARM USEWith the advent of genomic sequencing and high throughput genetic methods such as qPCR, it is now possible to obtain high-resolution enumeration of harmful algae from environmental samples. Costs for this are now lower, and results can be obtained within hours (or less), or even obtained on site. • Conduct a lab-based study to develop and implement a state of the art, rapid qPCR assay (including QA/QC) for Dinophysis species causing DSTs in Australian shellfish. This assay, similar to that currently used on site by Spring Bay Seafoods for the detection of Alexandrium (Paralytic Shellfish Toxins), will calculate the potential cell concentrations in environmental samples thereby providing real time (early) detection and monitoring of Dinophysis blooms.• Implement and field-test the qPCR assay for on-site use at Spring Bay Seafoods, Tasmania.
5. OBJECTIVE 5. PROVIDE COST VERSUS BENEFIT ANALYSIS OF IMPROVED TESTING OF DSTS IN TASMANIAN SHELLFISH• Conduct an economic impact analysis to estimate the reduction/annual savings in monitoring costs for regulatory authorities in Tasmania by using the most appropriate diagnostic testing for DSTs.• Calculate the reduction in commercial loss and economic impact from potentially harmful DST blooms in Tasmania following the introduction of the most appropriate diagnostic testing.
6. OBJECTIVE 6. CONDUCT A WORKSHOP TO TRAIN INTERESTED SHELLFISH INDUSTRY MEMBERS (TAS) IN THE USE OF RAPID TEST METHODS FOR DINOPHYSIS DETECTION AND DST TEST KITS IN ENVIRONMENTAL SAMPLESAMENDED:• Conduct a workshop to train shellfish industry members (Tas) in the use of the rapid method of qPCR for Dinophysis detection in environmental samples, and seek their advice and feedback on how to best move forward.ORIGINAL:• The project team will implement a training workshop to keep growers and industry abreast of qPCR methods, other research developments in DSTs, and seek their advice and feedback on how to best move forward.