Editorial: The European Food Risk Assessment Fellowship Programme (EU-FORA)
Bronzwaer, S., Le Gourierec, N., & Koulouris, S. (2016). Editorial: The European Food Risk Assessment Fellowship Programme (EU-FORA). EFSA Journal, 14(11).
Scientific reports of the Fellows
Published in special issues of the EFSA Journal
2022
Barbieri M V; Rodrigues A C; Febbraio F
Monitoring of pesticide amount in water and drinkable food by a fluorescence-based biosensor Journal Article
In: EFSA Journal, vol. 20, no. S1, pp. e200403, 2022.
Abstract | Links | Tags: biosensor, environmental monitoring, esterase-2, fluorescence, organophosphate pesticides, thermostable enzyme
@article{https://doi.org/10.2903/j.efsa.2022.e200403,
title = {Monitoring of pesticide amount in water and drinkable food by a fluorescence-based biosensor},
author = {Maria Vittoria Barbieri and Andreia CM Rodrigues and Ferdinando Febbraio},
url = {https://efsa.onlinelibrary.wiley.com/doi/abs/10.2903/j.efsa.2022.e200403},
doi = {https://doi.org/10.2903/j.efsa.2022.e200403},
year = {2022},
date = {2022-01-01},
journal = {EFSA Journal},
volume = {20},
number = {S1},
pages = {e200403},
abstract = {Abstract The identification of pollutants is crucial to protect water resources and ensure food safety. The available analytical methodologies allow reliable detection of organic pollutants such as pesticides; however, there is the need for faster, direct and continuous methodologies for real-time monitoring of pesticides. Fluorescent-based biosensors have been recently proposed as a valid alternative due to their advantage of being easy, cheap and specific. In this context, the aim of the present EU-FORA fellowship programme was to develop and apply a fluorescence-based biosensing device for the detection of organophosphate (OP) pesticides in water samples and drinkable food. The study was addressed using a mutant of the thermostable esterase-2 from Alicyclobacillus acidocaldarius (EST2-S35C) as a bioreceptor for OP pesticides. The use of EST2 involves some significant advantages including specificity and affinity towards OPs, and high stability over time in a different range of temperatures and pH. The protein was labelled to the fluorescent probe IAEDANS and fluorescence measurements of quenching in solution and in immobilised form were performed. The results showed good stability and sensitivity, reaching low limits of detection and quantification and a constant signal intensity over time. The addition of paraoxon quenched the fluorescence of the complex, reaching a plateau at 100 pmol paraoxon. The decrease of enzymatic activity of EST2-S35C-IAEDANS in the presence of paraoxon correlated the inhibition of the labelled enzyme with the decrease in fluorescence. The results from the application of the biosensor with real samples showed a decrease in fluorescence in surface water samples, contaminated by OPs. The use of the developed fluorescence-based biosensor demonstrated its applicability for real samples monitoring and could ensure the production of large amounts of data in a short period of time which can be used to address environmental and food safety risk assessment.},
keywords = {biosensor, environmental monitoring, esterase-2, fluorescence, organophosphate pesticides, thermostable enzyme},
pubstate = {published},
tppubtype = {article}
}
Abstract The identification of pollutants is crucial to protect water resources and ensure food safety. The available analytical methodologies allow reliable detection of organic pollutants such as pesticides; however, there is the need for faster, direct and continuous methodologies for real-time monitoring of pesticides. Fluorescent-based biosensors have been recently proposed as a valid alternative due to their advantage of being easy, cheap and specific. In this context, the aim of the present EU-FORA fellowship programme was to develop and apply a fluorescence-based biosensing device for the detection of organophosphate (OP) pesticides in water samples and drinkable food. The study was addressed using a mutant of the thermostable esterase-2 from Alicyclobacillus acidocaldarius (EST2-S35C) as a bioreceptor for OP pesticides. The use of EST2 involves some significant advantages including specificity and affinity towards OPs, and high stability over time in a different range of temperatures and pH. The protein was labelled to the fluorescent probe IAEDANS and fluorescence measurements of quenching in solution and in immobilised form were performed. The results showed good stability and sensitivity, reaching low limits of detection and quantification and a constant signal intensity over time. The addition of paraoxon quenched the fluorescence of the complex, reaching a plateau at 100 pmol paraoxon. The decrease of enzymatic activity of EST2-S35C-IAEDANS in the presence of paraoxon correlated the inhibition of the labelled enzyme with the decrease in fluorescence. The results from the application of the biosensor with real samples showed a decrease in fluorescence in surface water samples, contaminated by OPs. The use of the developed fluorescence-based biosensor demonstrated its applicability for real samples monitoring and could ensure the production of large amounts of data in a short period of time which can be used to address environmental and food safety risk assessment.
Rodrigues A C; Barbieri M V; Febbraio F
Monitoring of pesticide amount in fruit and vegetables by a fluorescence-based sensor Journal Article
In: EFSA Journal, vol. 20, no. S1, pp. e200419, 2022.
Abstract | Links | Tags: environmental monitoring, enzymatic bioreceptor, fluorescence-based methods, organophosphate pesticides, thermophilic esterase
@article{https://doi.org/10.2903/j.efsa.2022.e200419,
title = {Monitoring of pesticide amount in fruit and vegetables by a fluorescence-based sensor},
author = {Andreia CM Rodrigues and Maria Vittoria Barbieri and Ferdinando Febbraio},
url = {https://efsa.onlinelibrary.wiley.com/doi/abs/10.2903/j.efsa.2022.e200419},
doi = {https://doi.org/10.2903/j.efsa.2022.e200419},
year = {2022},
date = {2022-01-01},
journal = {EFSA Journal},
volume = {20},
number = {S1},
pages = {e200419},
abstract = {Abstract Faster, sensitive and real-time methods for detecting organophosphate (OP) pesticides are urged for in situ monitoring of these widely spread contaminants. For this reason, several efforts have been addressed for the development of performant biosensors. The thermostable enzyme esterase-2 from Alicyclobacillus acidocaldarius (EST2), with a lipase-like Ser-His-Asp catalytic triad with a high affinity to OPs, is a promising candidate as a bioreceptor for biosensor development. Within this EU-FORA fellowship project, two different components of the biosensor were evaluated: (i) the use of the enzymatic bioreceptor in solution or immobilised in a solid membrane; (ii) the measurement of fluorescence quenching by direct measurement of the fluorescence probe intensity signal or by fluorescence resonance energy transfer (FRET) from the tryptophans located in the catalytic site of the enzyme to a binded fluorescence probe. Fluorescence spectroscopy is among the most used techniques in analytical chemistry laboratories, mainly due to its high sensitivity and simplicity. To this aim, the developed IAEDANS-labelled EST2-S35C mutant has been used. Fluorometric measurements with both methods showed linearity with increased EST2-S35C concentrations. No significant interference on FRET measurements was observed due to changes in medium pH or due to the addition of other organic components (glucose, ascorbic acid, yeast extract). Both methods presented similar sensitivity towards detecting OPs, with fluorescence quenching due to the presence of paraoxon at environmentally relevant concentrations from 0.09 µM. The obtained results are of high relevance to further development of biosensors for the pesticide monitoring that: (i) decrease the expenses of the analysis; (ii) simplify the procedures for pesticide detection; (iii) reduce the time of response. Furthermore, the use of biosensors for pesticides real-time and in situ detection of pesticides promises to increase the number of samples analysed, providing a larger amount of data for food safety risk assessment.},
keywords = {environmental monitoring, enzymatic bioreceptor, fluorescence-based methods, organophosphate pesticides, thermophilic esterase},
pubstate = {published},
tppubtype = {article}
}
Abstract Faster, sensitive and real-time methods for detecting organophosphate (OP) pesticides are urged for in situ monitoring of these widely spread contaminants. For this reason, several efforts have been addressed for the development of performant biosensors. The thermostable enzyme esterase-2 from Alicyclobacillus acidocaldarius (EST2), with a lipase-like Ser-His-Asp catalytic triad with a high affinity to OPs, is a promising candidate as a bioreceptor for biosensor development. Within this EU-FORA fellowship project, two different components of the biosensor were evaluated: (i) the use of the enzymatic bioreceptor in solution or immobilised in a solid membrane; (ii) the measurement of fluorescence quenching by direct measurement of the fluorescence probe intensity signal or by fluorescence resonance energy transfer (FRET) from the tryptophans located in the catalytic site of the enzyme to a binded fluorescence probe. Fluorescence spectroscopy is among the most used techniques in analytical chemistry laboratories, mainly due to its high sensitivity and simplicity. To this aim, the developed IAEDANS-labelled EST2-S35C mutant has been used. Fluorometric measurements with both methods showed linearity with increased EST2-S35C concentrations. No significant interference on FRET measurements was observed due to changes in medium pH or due to the addition of other organic components (glucose, ascorbic acid, yeast extract). Both methods presented similar sensitivity towards detecting OPs, with fluorescence quenching due to the presence of paraoxon at environmentally relevant concentrations from 0.09 µM. The obtained results are of high relevance to further development of biosensors for the pesticide monitoring that: (i) decrease the expenses of the analysis; (ii) simplify the procedures for pesticide detection; (iii) reduce the time of response. Furthermore, the use of biosensors for pesticides real-time and in situ detection of pesticides promises to increase the number of samples analysed, providing a larger amount of data for food safety risk assessment.
2018
Rusko J; Febbraio F
Development of an automated multienzymatic biosensor for risk assessment of pesticide contamination in water and food Journal Article
In: EFSA Journal, vol. 16, no. S1, pp. e16084, 2018.
Abstract | Links | Tags: biosensing device, environmental monitoring, organophosphate pesticides, thermophilic esterase
@article{https://doi.org/10.2903/j.efsa.2018.e16084,
title = {Development of an automated multienzymatic biosensor for risk assessment of pesticide contamination in water and food},
author = {Janis Rusko and Ferdinando Febbraio},
url = {https://efsa.onlinelibrary.wiley.com/doi/abs/10.2903/j.efsa.2018.e16084},
doi = {https://doi.org/10.2903/j.efsa.2018.e16084},
year = {2018},
date = {2018-01-01},
journal = {EFSA Journal},
volume = {16},
number = {S1},
pages = {e16084},
abstract = {Abstract The goal of this research is to better address the problems related to the widespread presence of pesticides in the environment. Despite the unquestionable utility of the pesticides against various pests in the agricultural field, most pesticides and the corresponding pesticide residues are toxic to the environment and hazardous to human health. The recent literature on organophosphate compounds emphasises a clear correlation between their use and the occurrence of disorders in the nervous system, especially in children. The conventional systems for the detection and analysis of these compounds are expensive, time-consuming and require highly specialised operators; moreover, no online automated screening systems are yet available, that would allow the identification and quantification of the presence of these chemicals in samples from industrial sectors such as the food industry. Esterase-based biosensors represent a viable alternative to this problem. In this fellowship programme, we aim to develop a robust and sensitive methodology that enables the screening of toxic compounds using a streamlined process, using an automated robotic system to achieve a continuous monitoring for risk assessment of pesticides.},
keywords = {biosensing device, environmental monitoring, organophosphate pesticides, thermophilic esterase},
pubstate = {published},
tppubtype = {article}
}
Abstract The goal of this research is to better address the problems related to the widespread presence of pesticides in the environment. Despite the unquestionable utility of the pesticides against various pests in the agricultural field, most pesticides and the corresponding pesticide residues are toxic to the environment and hazardous to human health. The recent literature on organophosphate compounds emphasises a clear correlation between their use and the occurrence of disorders in the nervous system, especially in children. The conventional systems for the detection and analysis of these compounds are expensive, time-consuming and require highly specialised operators; moreover, no online automated screening systems are yet available, that would allow the identification and quantification of the presence of these chemicals in samples from industrial sectors such as the food industry. Esterase-based biosensors represent a viable alternative to this problem. In this fellowship programme, we aim to develop a robust and sensitive methodology that enables the screening of toxic compounds using a streamlined process, using an automated robotic system to achieve a continuous monitoring for risk assessment of pesticides.