Biomaterial culture conditions impacting the performance of a PAM fluorometry based aquatic phytotoxicity assay

An aquatic phytotoxicity assay, based on the principles of pulse amplitude modulated (PAM) fluorometry has recently been developed and validated under laboratory conditions. Characteristics of the assay include the use of photosynthesising biomaterial, most frequently whole organism microalgae. The instrument employs light probing measurements to monitor chlorophyll fluorescence signals emitted by the biomaterial component. These characteristics could leave assay performance susceptible to interference by minor variations in biomaterial treatment and culture conditions prior to testing. This study investigates assay performance in response to variations in two microalgae culture parameters; short-term light history (24h) prior to testing and the sterility of long-term culture conditions. Light history of the four microalgal species tested significantly impacted their toxicity response, as measured with the assay. Light treatments of 5 micromol photons m(-2)s(-1) produced the highest photosystem II quantum yields (Phi(II)) whilst higher light intensities resulted in an inverse relationship between Phi(II) and the measured toxicity response (inhibition (%) of photochemistry). Of the two microalgal cultures tested, sterility of culture conditions significantly impacted the performance of the green freshwater algae, Chlorella vulgaris as assay biomaterial. On average 1 microg L(-1) diuron inhibited photochemistry 2.6% less in axenically cultured C. vulgaris compared with non-axenically maintained cultures. This investigation series contributes valuable quality assurance data towards microalgal based PAM fluorometry assays and emphasises the importance of such investigations if new biorecognition systems are to be accredited and/or routinely incorporated for biomonitoring purposes.