Microalgae dual-head biosensors for selective detection of herbicides with fiber-optic luminescent O2 transduction.

The microalgal species Dictyosphaerium chlorelloides (D. c.) was immobilized into porous silicone films and their photosynthetic activity was monitored with an integrated robust luminescent O2 sensor. The biosensor specificity towards a particular pesticide has been achieved by manufacturing a fiber-optic dual-head device containing both analyte-sensitive and analyte-resistant D. c. strains. The latter are not genetically modified microalgae, but a product of modified Luria-Delbrück fluctuation analysis followed by ratchet selection cycles. In this way the target herbicide decreases the O2 production of the analyte-sensitive immobilized strain without affecting the analyte-resistant population response; any other pollutant will lower the O2 production of both strains. The effect of the sample flow-rate, exposure time to the herbicide, biomass loading, biosensor film thickness, intensity of the actinic light, illumination cycle, and temperature on the biosensor response has been evaluated using waterborne simazine as test bench. The biosensing device is able to provide in situ measurements of the herbicide concentration every 180 min. The biosensor limit of detection for this herbicide was 12 µg L(-1), with a working range of 50-800 µg L(-1). The biosensor specificity to simazine has been assessed by comparing its response to that of isoproturon.

Toxicity and adaptation of Dictyosphaerium chlorelloides to extreme chromium contamination.

Metals are often spilled by industries into inland water environments, with adverse consequences. Numerous papers have reported that heavy metals produce massive destruction of algae. Nevertheless, algal populations seem to become tolerant when they have had previous exposures to heavy metals. Because the mechanisms allowing heavy metal tolerance of algae are not yet known, the present study analyzed the effect of hexavalent chromium on growth and photosynthetic performance of Dictyosphaerium chlorelloides, stressing on the adaptation mechanisms to chromium contamination. Growth and photosynthetic performance of algal cells were inhibited by Cr(VI) at 10 mg/L, and the 72-h median inhibition concentration was established as 1.64 and 1.54 mg/L, respectively. However, after further incubation for a three month period in an environment with 25 mg/L of chromium, some rare, chromium-resistant cells occasionally were found. A Luria-Delbrück fluctuation analysis was performed to distinguish between resistant algae arising from rare, spontaneous mutations and resistant algae arising from physiological adaptation and other adaptive mechanisms. Resistant cells arose only by spontaneous mutations before the addition of chromium, with a rate of 1.77 x 10(-6) mutants per cell division. From a practical point of view, the use of both chromium-sensitive and chromium-resistant genotypes could make possible a specific algal biosensor for chromium.