Microalgae-Based Fluorimetric Bioassays for Studying Interferences on Photosynthesis Induced by Environmentally Relevant Concentrations of the Herbicide Diuron.

The widespread agricultural use of the phenylurea herbicide Diuron (DCMU) requires the investigation of ecotoxicological risk in freshwater and soil ecosystems in light of potential effects on non-target primary producers and a heavier effect on higher trophic levels. We used microalgae-based fluorimetric bioassays for studying the interferences on the photosynthesis of a freshwater and soil model green microalga (Chlamydomonas reinhardtii) induced by environmentally relevant concentrations of the herbicide DCMU. Measurements of steady-state chlorophyll a (Chl-a) fluorescence emission spectra were performed; as well, the kinetics of the Chl-a fluorescence transient were recorded. Percentage indexes of interference on photosynthesis were calculated after comparison of steady-state and kinetic Chl-a fluorescence measurements of DCMU-exposed and control C. reinhardtii cell suspensions. The results obtained after 30 min exposure to the herbicide DCMU confirmed a significant inhibitory effect of DCMU 2 µg/L, and no significant differences between %ι values for DCMU 0.2 µg/L and 0.02 µg/L exposures. Positive %ε values from kinetic measurements of the Chl-a fluorescence transient confirmed the same interfering effect of 2 µg/L DCMU on PSII photochemistry in the exposed C. reinhardtii cell suspensions. Negative values of %ε observed for 0.2 and 0.02 µg/L DCMU exposures could be attributable to a presumptive 'stimulatory-like' effect in the photochemistry of photosynthesis. Short-term exposure to sub-µg/L DCMU concentration (≤0.2 µg/L) affects the photosynthetic process of the model microalga C. reinhardtii. Similar environmental exposures could affect natural communities of unicellular autotrophs, with hardly predictable cascading secondary effects on higher trophic levels.

Towards Simazine Monitoring in Agro-Zootechnical Productions: A Yeast Cell Bioprobe for Real Samples Screening.

Simazine is an herbicide that is able to contaminate surface waters, ground waters, and milk/dairy products, thus posing concerns in both environmental health and food safety. A yeast-based bioprobe was utilized to detect simazine in spiked real samples of livestock drinking water and raw cow's milk. Yeast aerobic respiration was taken as short-term toxicological endpoint. We carried out comparative measures of yeast oxygen consumption between simazine-spiked samples and blank samples. Percentage interference (%ρ) on yeast aerobic respiration was calculated through the comparison of aerobic respiration of simazine-exposed and non-exposed yeast cells. The method was optimized for raw cow's milk samples by using boric acid as fungistatic agent in order to avoid cellular proliferation. Overall, the results have shown that simazine can be detected up to concentrations five times below the EU legal concentration limits for drinking water (0.02 ppb) and cow's milk (2 ppb) (%ρ values of 18.53% and 20.43% respectively; %RSD ≤ 15%). Dose-effect relationships of simazine were assessed. The findings of the bioassays match reasonably well with known mechanisms of toxicity and intracellular detoxification in yeast. A correlation between fat content in milk samples and analytical performance of the bioprobe was established. Results suggest the involvement of a matrix effect, presumably due to lipid sequestration of simazine. The yeast-based bioprobe has proved to be sensitive and suitable for the detection of simazine in real samples in concentrations of interest.