In this project, marine water collected from the HAB in South Australia will be sent to the PI at the University of Technology Sydney, and will be used to isolate and culture strains of Karenia species into clean, unialgal cultures. Samples from the HAB previously sent to the PI over the past month will be examined, and if Karenia is present and still alive, it will be isolated into uni-algal culture. Cultured strains will be identified using molecular genetic methods.

The purpose of obtaining unialgal strains of Karenia species from this HAB is:
- The identification of the species responsible for toxic effects and brevetoxin production;
- To allow for the development of rapid, early warning molecular detection assays such as qPCR (ie similar to COVID detection);
- To enable future toxin uptake rates and mechanisms into commercial seafood to be experimentally examined, to understand brevetoxin threats to seafood.

Timely action is essential, as the duration of the HAB in South Australia is unpredictable, and the opportunity to collect representative samples may be brief. Microalgal samples commonly die after periods of weeks, even if kept at constant temperature and light conditions, and provided with the optimal media for growth. The only means to maintain Karenia strains alive over the long term, is to isolate and purify them into culture conditions in which they can be maintained indefinitely.

There are few people in Australia with the skills to isolate new strains of single-celled marine algae to allow them to grow in culture. The work involved is painstaking and time consuming. Hence, the research described in this proposal is both highly specialised and time critical. It is necessary in order that a full analysis of the reasons for and impacts of the South Australian HAB can be understood, and risks from it be minimised for all seafood industries in Australia in future.

This work does not duplicate other research or sampling conducted to date on the HAB. To date, no attempts at Karenia strain isolation have occurred elsewhere in Australia. This project complements work by SARDI/PIRSA who are documenting the extent of chlorophyll a in the waters off South Australia using remote sensing, as a proxy marker, (although chlorophyll a includes other microalgae, not only Karenia); and the work of Clinton Wilkinson from the South Australian Shellfish Quality Assurance Advisory Committee (SASQAAC), who are conducting routine brevetoxin testing on South Australian commercial oysters and routine microalgal cell counts on water samples. Other marine scientists are collecting dead fish and documenting marine ecosystem impacts of the HAB.