Fast pesticide pre-screening in marine environment using a green microalgae-based optical bioassay.

The present study evaluates an optical bioassay based on green photosynthetic microalgae as a promising alternative for monitoring of relevant seawater pollutants. Photosystem II fluorescence parameters from several microalgae species were examined in the presence of three common marine pesticides that act as photosynthesis inhibitors. The three pollutants were detected within 10 min in concentrations between ng/L-µg/L. The different algae species showed slightly diverse pesticide sensitivities, being Chlorella mirabilis the most sensitive one. Potential interferences due to oil-spill pollutants were discarded. The lipid content was characterized to identify microorganisms with suitable mechanisms that could facilitate stress acclimatization. C. mirabilis presented elevated content of unsaturated lipids, showing a promising potential for biosensing in saline stress conditions. The optimized microalgae-based bioassay was preliminarily incorporated into a marine buoy for autonomous pre-screening of pesticides in coastal areas, demonstrating its suitability for real-time monitoring of marine water and quantitative evaluation of total biotoxicity.

Towards an integrated biosensor array for simultaneous and rapid multi-analysis of endocrine disrupting chemicals.

In this paper we propose the construction and application of a portable multi-purpose biosensor array for the simultaneous detection of a wide range of endocrine disruptor chemicals (EDCs), based on the recognition operated by various enzymes and microorganisms. The developed biosensor combines both electrochemical and optical transduction systems, in order to increase the number of chemical species which can be monitored. Considering to the maximum residue level (MRL) of contaminants established by the European Commission, the biosensor system was able to detect most of the chemicals analysed with very high sensitivity. In particular, atrazine and diuron were detected with a limit of detection of 0.5nM, with an RSD% less than 5%; paraoxon and chlorpyrifos were revealed with a detection of 5 µM and 4.5 µM, respectively, with an RSD% less than 6%; catechol and bisphenol A were identified with a limit of detection of 1 µM and 35 µM respectively, with an RSD% less than 5%.

New platform of biosensors for prescreening of pesticide residues to support laboratory analyses.

Millions of tons of pesticides are applied worldwide annually in agriculture. Among them, herbicides such as triazines and ureas, originating from agricultural runoff, can contaminate soils and surface and ground waters with severe toxic effects on humans. Nowadays, different analytical techniques are available for the detection of these chemicals; however, most of them are expensive and time-consuming, especially in the case of routine analyses. For this reason, on the basis of results collected through many years of experience in the field of photosynthetic organisms, we designed a biosensor platform intended for the easy, low-cost, and fast prescreening of photosynthetic herbicides. The platform combines the possibilities of amperometric and optical transduction systems, which have proven to be highly sensitive (limits of detection = 10(-10)-10(-8) M). The use of genetically modified algae strengthens the power of the platform, allowing different subclasses of herbicides to be recognized. The system has been validated for the analysis of environmental water and is proposed to support laboratories involved in the control of water pollution.

Optical biosensors for environmental monitoring based on computational and biotechnological tools for engineering the photosynthetic D1 protein of Chlamydomonas reinhardtii.

Homology-based protein modelling and computational screening followed by virtual mutagenesis analyses were used to identify functional amino acids in the D1 protein of the photosynthetic electron transfer chain interacting with herbicides. A library of functional mutations in the unicellular green alga Chlamydomonas reinhardtii for preparing biomediators was built and their interactions with herbicides were calculated. D1 proteins giving the lowest and highest binding energy with herbicides were considered as suitable for preparing the environmental biosensors for detecting specific herbicide classes. Arising from the results of theoretical calculations, three mutants were prepared by site-directed mutagenesis and characterized by fluorescence analysis. Their adsorption and selective recognition ability were studied by an equilibrium-adsorption method. The S268C and S264K biomediators showed high sensitivity and resistance, respectively, to both triazine and urea classes of herbicides. When immobilized on a silicon septum, the biomediators were found to be highly stable, remaining so for at least 1-month at room temperature. The fluorescence properties were exploited and a reusable and portable multiarray optical biosensor for environmental monitoring was developed with limits of detection between 0.8 x 10(-11) and 3.0 x 10(-9), depending on the target analyte. In addition, biomediator regeneration without obvious deterioration in performance was demonstrated.