Research activities of the Fougères Laboratory
The research activities of the Fougères Laboratory focus on several themes, aimed at better understanding and characterising the various points in the chain of events leading from exposure to one or more substances to the occurrence of adverse effects. More specifically, the laboratory works to improve knowledge on the benefits and risks related to the food industry’s use of veterinary medicinal products and disinfectants and the analysis of the effects of antimicrobial agents (antibiotics and biocides). More broadly, it is involved in characterising the toxicological hazards associated with emerging substances and contaminants.
Disinfectant biocides - antimicrobial resistance
- QESABIO (2019 - 2020)
Quantitative assessment of variability in Salmonella excretion, and impact of biocide use on the development of antimicrobial resistance after cleaning/disinfection, on pig farms
This project was launched to address two concerns in the pig sector: bacterial resistance to antibiotics and food safety. Coordinated by the Antibiotics, Biocides, Residues and Resistance Unit, it also involved ANSES's Ploufragan-Plouzané-Niort Laboratory and Regulated Products Assessment Department. The results showed that biocidal products containing quaternary ammoniums and glutaraldehyde applied during cleaning and disinfection operations led to an approximate 1,000-fold reduction in total flora and the quantity of E. coli. Exposure to these biocides did not cause E. coli to develop resistance to four target antibiotics or to biocidal substances representative of the main chemical classes used in this sector. The level of Salmonella excretion by slaughter pigs was relatively low with two-thirds of samples having less than 10 Salmonella per gram of faeces.
- IMPART (2018 - 2023)
Improving phenotypic antimicrobial resistance testing
Funding: European One Health EJP Programme
As part of Work package 1 of the IMPART project, the Antibiotics, Biocides, Residues and Resistance Unit was in charge of developing, improving and validating a screening method for the detection, characterisation and confirmation of colistin resistance in Enterobacteriaceae sampled from animals on farms and from their meat at distribution. The unit organised several ring tests, in which the project’s 11 European partners participated, to evaluate the method developed and also the method proposed as part of the second work package, for the detection of carbapenemase-producing Enterobacteriaceae. The analysis of results showed that further tests are necessary to improve the performance of the methods. Reports and presentations relating to this work are available on the website of the One Health EJP and on the Zenodo platform.
- Ocean-15 project
Funding: French National Research Agency (ANR)
The ANR Ocean-15 project, which had four partners and was coordinated by National University of Ireland Galway, lasted five years. It strove to better understand the behaviour of a toxic microalga, Ostreopsis cf. ovata, found in the Mediterranean. It also identified the compounds produced and their role in the toxic effects observed in humans. Ostreopsis secretes toxins called ovatoxins that are analogues of a well-known toxin known as palytoxin. Effects reported in humans involving the skin, eyes, ears, nose and larynx have been associated with Ostreopsis blooms. Although these toxins are also found in some seafood products, no cases of human poisoning related to their consumption have been reported. The project's participants purified two variants (ovatoxins a and d) from Ostreopsis cultures; the Fougères Laboratory tested their toxicity using various cell models. Both ovatoxins were found to be toxic to cells of the nervous system and to lung cells, with no major differences between the variants and at levels similar to those observed with palytoxin. However, in human cultures mimicking the intestinal barrier, both toxins were less toxic than palytoxin.
Veterinary medicinal product residues
- Evaluation and validation of nitrofuran and colistin ELISA kits (2021 - 2022)
Funding: European Commission
As part of its reference missions, the laboratory evaluated and validated the technical performance of several enzyme-linked immunosorbent assay (ELISA) kits. Three of the four tested kits for the detection of residues of nitrofuran metabolites (prohibited substances) in aquaculture products (shrimp, fish) showed detection capacities below the current European regulatory requirements (1 µg/kg) with a false-positive rate of around 10%. For colistin, an antibiotic primarily used in pigs and poultry to treat gastrointestinal infections that is included on the list of highest priority critically important antimicrobials of the World Health Organization, three kits were evaluated and validated. They were all highly specific (no false positives) for the detection of colistin in pig and poultry muscle. However, only one of the kits showed a truly satisfactory detection capacity (30 µg/kg) in relation to the maximum residue limit (150 µg/kg). These findings were highlighted in publications in European peer-reviewed journals.
- Detection of dye residue markers (2021 - 2022)
In aquaculture, the use of dye compounds in the class of triarylmethanes, which have antiseptic and antifungal properties, is prohibited because of their toxicity. Little is known about the fate of some of these compounds in the body. It was therefore necessary to carry out work to define residue markers that should be screened for in foodstuffs of animal origin for surveillance purposes. Thesis work explored various approaches for studying the metabolism of one of these dyes, Victoria Pure Blue BO. The results of in vitro and in vivo metabolism studies were combined to propose a direct metabolite arising from exposure to Victoria Pure Blue as an appropriate residue marker in salmonids. Biological disruption in terms of the production of bile acids was also observed, after trout were exposed to two triarylmethanes. This work was undertaken in collaboration with the Faculty of Fisheries and Protection of Waters in the Czech Republic and the Ploufragan-Plouzané-Niort Laboratory for animal experimentation in fish. The results were described in three scientific publications.
- SILVERPROTECT (2020 - 2022)
Innovative materials including silver ions to guarantee the safety of aquaculture products
Funding: European Maritime and Fisheries Fund
This project is coordinated by the Boulogne-sur-Mer site of ANSES’s Laboratory for Food Safety. Its objective is to assess the performance of antimicrobial materials covering fish preparation surfaces. It is being carried out in collaboration with the aquaculture industry, the PureZone consortium, the University Institute of Technology (IUT) in Saint-Brieuc, and the Technical Centre for Agri-Food Expertise in Saint-Lô.
- QAC-BIoM (2021 - 2023)
Quaternary ammonium compounds – bowel-intestinal-microbiotal
Preliminary exploratory study investigating the impact of quaternary ammoniums on permeability and the microbiota, consequences for chronic inflammatory bowel diseases. For the first time, the interplay between exposure to low concentrations of disinfectant biocides, the gut microbiota and the intestinal barrier will be studied to expand knowledge on chronic inflammatory diseases using a holistic interdisciplinary approach. This project is coordinated by the Infinite unit (Inserm - University of Lille - Lille University Hospital) and involves three units of the Fougères Laboratory (ARC, EMAD, AB2R).
- OSABt (2019 - 2021)
Monitoring the risk of food poisoning associated with the use of Bt bio-insecticides
Bacillus thuringiensis (Bt) is number one on the global market of microbial bio-insecticides and populations can be exposed to it via food. However, it belongs to the same bacterial group as Bacillus cereus, which has been associated with several food poisoning outbreaks in humans. The aim of this project is to generate a better understanding of the risks related to the use of bio-insecticides containing Bacillus thuringiensis.
- Tentacles (2020 - 2024)
Temperature-responsive nanogels for targeted delivery of microRNAs in wound healing and tissue regeneration applications
Funding: Horizon 2020 programme
This project’s objective is to develop an innovative nanogel, containing iron oxide nanoparticles and targeted microRNAs, within a polymer for skin treatments and tissue regeneration. Iron oxide promotes the induction of fibrogenesis and microRNAs are involved in modulating the expression of specific genes. Various nanogels will be synthesised. Their toxicity and genotoxicity will be evaluated in vivo and in vitro.
- CONTALIM (2020 - 2023)
Food contamination by antibiotics
Funding: Brittany region and ANSES
The cross-contamination of animal feed by antibiotics, resulting from the use of the same production lines for all types of feed, including medicated feedingstuffs, was demonstrated by surveillance plans implemented on French farms in 2017 and 2018. A thesis under way is assessing the risk of residues being transferred to animal feed and also the risk of resistance emerging in the microbiota of pigs given feed contaminated by the antibiotics most commonly found during surveillance plans.
- Antibiotrace (2019 - 2022)
PBPK methodologies for predicting concentrations of antibiotics in tissue based on trace amounts found in milk, in several species (cows, sheep and goats) and with several antibiotics
Funding: Nouvelle-Aquitaine Region and ANSES
This project concerns the evaluation of antimicrobial residues in milk. The presence of residues promotes the emergence and development of resistant bacteria. However, the studies undertaken face various challenges, especially from an analytical standpoint. They are long, complex and expensive. They could benefit from recent progress in the field of physiologically based pharmacokinetic (PBPK) modelling. This project, conducted in collaboration with internal (Ploufragan-Plouzané-Niort Laboratory) and external (Inserm, University of Poitiers, the company Ceva Santé Animale, and the agricultural secondary schools of Melle and Venours) partners, focused on three milk-producing animal species. Residue levels in milk are currently being analysed.
- Capbiola (2021 - 2023)
Development of electrochemical and colorimetric biosensors for the detection of disinfectant biocide residues in the dairy industry
Funding: Brittany Region
Optical biosensors are used to detect a wide variety of food contaminants because they have the best characteristics (sensitivity, speed, low cost, high throughput) for field use. Already evaluated as part of the laboratory’s reference missions for the detection of antimicrobial residues, they could be a promising alternative for the detection of biocide residues. This detection could rely on rinses from operations to clean and disinfect food contact surfaces. The Capbiota project is funded by the Brittany Region as part of a Sustainable Attractiveness Strategy call to recruit a PhD researcher for 18 months. It will deal with the development, optimisation, evaluation and validation of optical (colorimetric and fluorimetric) and electrochemical biosensors, for the detection of quaternary ammonium compounds and amines in the dairy industry. In addition to drawing from the unit's expertise relating to the detection of antimicrobial and veterinary medicinal product residues, the project will benefit from the scientific and technical capacities of a laboratory of the University of Perpignan (Biosensors - Analysis - Environment (BAE)).