A Temperature-Controlled Fluorescence Fingerprint for Identifying Pseudo-nitzschia hasleana in Harmful Algal Blooms

Harmful algal blooms (HABs) caused by toxic species such as Pseudo-nitzschia hasleana pose significant risks to marine ecosystems and human health. This study investigates the effects of heating rate on the fluorescence temperature curves (FTCs) of P. hasleana and compares them with non-toxic species (Phaeodactylum tricornutum and Picochlorum maculatum) to design a reliable detection method. An increasing heating rate leads to a change in the temperature spectrum of the fluorescence of the studied algae and to increasing differences between them. During the study, the FTCs were measured in the temperature range of 20–80 °C and at heating rates of 1, 2, 3, and 6°/min. The results showed that P. hasleana exhibited a distinct local fluorescence maximum at 45–55 °C when heated at a rate of 3 °C/min or more, which was absent in non-toxic species. Additionally, rapid heating (6 °C/min) preserved fluorescent pigment–protein complexes, yielding four-fold higher fluorescence intensity at 70–80 °C compared to slower rates. There were no such changes for the microalgae P. maculatum and P. tricornutum. The results of this study make it possible to increase the efficiency of detecting hazardous microalgae using non-invasive optical monitoring methods. These findings demonstrate that controlled heating protocols can enhance the species-specific identification of toxic microalgae, offering a practical tool for early HAB detection.