Fast detection of contaminants and adulteration in raw milk using dip-stick nanophotonic sensors
About the Project
To ensure food quality and to safeguard public health, regulatory authorities have set specific maximum allowable concentrations and rigorous control procedures for substances considered as contaminants or agents of fraud in food. Nowadays, the determination of these substances requires complex, time consuming and costly laboratory analyzes. Nowadays, the determination of these substances requires complex, time consuming and costly laboratory analyzes. However, in the case of perishable foods, such as milk, immediate control of the raw material and processed products is required for their release on the market. Among the parameters required to be determined in milk, the most important are the presence of bacteria and Aflatoxin M1 (AFM1). as well as the adulteration of sheep's or goat's milk with cow's milk. Bacteria and AFM1 affect all types of milk. Counterfeiting mainly concerns the protection of consumers with an allergic predisposition to cow's milk, but also the detection of fraud involving the mixing of sheep's or goat's milk products with cow's milk for profit. For bacteria, the applied detection methods require enrichment of the sample for at least 24 hours while the determination of AFM1 and the detection of fraud is carried out with costly laboratory analyzes. At FOODSENS a new system will be developed which will meet its urgent need food industry to compress the time between sampling and result but also the cost of analyzes. The proposed system will highlight the significant advantages of the simultaneous detection of AFM1 and adulteration of ovine and caprine animals with cow's milk at milk collection points and the detection of bacteria such as Bacillus cereus and Listeria monocytogenes by meeting the required legal limits.
Objectives of the project
The aim of the project is to develop a tool for rapid and low cost analyzes in the Food Sector. More specifically, the main objectives are the detection of AFM1 and adulteration of sheep and goats with cow's milk at its collection points and the detection of Bacillus cereus and Listeria monocytogenes in less than 10 min if the samples have loads> 100 cfu/mL and up to 6 hours for very low or zero loads. To achieve these goals, FOODSENS will take advantage of the experience and successful cooperation in the development of innovative biosensors of two research laboratories of NCSR "Demokritos" and the many years of experience of the Agricultural University of Athens in the detection of microorganisms in food. Their know-how will be combined with the capacity of a start-up SME (ThetaMetrisis) with intense export activity and specialization in technological solutions for photon systems, which will develop the measuring device. The evaluation of FOODSENS will be carried out by ΥIOTIS Food Industry. One of the main objectives of the proposal is to draw up a business plan for the commercial use of the FOODSENS device in the global market for analytical instruments. Finally, a special sub-action that will undertake the actions of promotion and dissemination of the results in both the academic and the business community. The actions of utilization and dissemination of the results will be coordinated by SEVT and the PRAXI network.
The disruptive FOODSENS tool is based on a directly-immersible photonic chip operating as a label-free biosensor. Its great innovation is that it uses integrated nanophoton circuits using wide-range Mach-Zehnder interferometers (BB-MZI). The light inside and outside the photon chip is coupled to external double-stranded fibers that connect the input to the white light source and the output to a digital spectral analyzer. The photon chip is self-aligning with the double-stranded fiber via a properly designed mechanical coupler, so that most of it protrudes as a submersible terminal. In each tile, three BB-MZI will be implemented, one of which will be the reference interferometer. The sensory arm of the BB-MZI reference will be coated with an inert protein while the sensory arms of the other two will be modified with specific recognition biomolecules for the simultaneous detection of AFM1 and bovine casein in goat's and sheep's milk, and the bacteria Bacillus cereus and Listeria monocytogenes. The integration into the reference BB-MZI chip provides the advantage that it automatically removes the effect of the sample matrix and allows the analyzers to be detected by a single self-referential measurement through direct contact of the sensor with the sample to be analyzed. The system will have the following competitive advantages over all existing analytical solutions: - Decoupling of the final system from microfluidic devices - Simplicity of operation, mechanical stability, cost reduction and system volume - Ability to simultaneously detect two analyzers without the use of tracers - Self-referentiality - Ability to implement the tile in conventional integrated circuit processing facilities This unique combination of advantages makes the proposed system very attractive for applications where reliability, speed and compression of analysis costs are required.
Expected results
The FOODSENS system aspires to offer the Greek and global Food Industry more effective assurance of the quality and safety of its products while it is a business opportunity for all stakeholders. The expected benefits can be summarized as follows: For the Greek Food Industry, which is one of the most important competitive and extroverted sectors of the secondary sector of the domestic economy, it is expected through the utilization of the results of FOODSENS to reduce production costs and increase its competitiveness. In addition, it is expected: empowerment of Greek food SMEs through the use of methodologies that do not require expensive equipment, creation of new jobs due to the expansion of their turnover, strengthening their export activity, improving consumer confidence in the products of the Greek Food Industry, of the most active research & development. In addition, for the Greek and European society there will be: - Increased guarantee for an even higher level of safety of the products produced - Increased protection of consumer health from foodborne diseases - Creation of new highly specialized jobs for young scientists and opportunities to participate in the implementation of cutting-edge technologies. For YIOTIS the results of the project will be immediately usable and will be able to further strengthen the already high level of quality control. In this context, the following are expected to be achieved: - Enhancing the already high level of safety and quality of its products - Reduction of quality control costs - Utilization of the system as a fast and reliable tool for detection of pathogenic microorganisms, aflatoxin M1 and adulteration of goat's and sheep's milk with cow's milk - Limiting the storage time of raw materials and products and more immediate placing of batches on the market - Strengthening the image of the company as a leader in the field of safety and quality control of its products - Strengthening its competitive position in the Greek market and strengthening its efforts to penetrate new markets abroad. For ThetaMetrisis, the area of bioanalytic systems is of particular interest due to the size of the specific market and the know-how that the company holds in the design / manufacture of photon systems. The company is expected to achieve penetration in the market of analytical tools for the Food Industry and increase its turnover, increase its human resources with highly qualified personnel and the development of other similar analytical systems for the Food Industry. For SEVT and the PRAXI network, the expected results of the project do not concern only the participating companies but the Food Industry as a whole. In this context, SEVT is expected to utilize the knowledge generated to provide its members with upgraded services and information on new rapid analysis techniques, to open a dialogue with the competent state bodies on the use of modern analysis techniques, and to strengthen the promotion actions of innovation. that develops. In a more general context, the consortium aims to strengthen the domestic food industry and to offer to Greek society as follows: For the academic institutions involved are expected: - Acquisition of new knowledge and extension of existing know-how in new fields related to the needs of the domestic industry - Strengthening the academic profile by increasing publications / announcements - Contribute to stopping the brain drain with jobs for young people in cutting-edge science / technology - Expanding their research fields and strengthening relations with the Food Industry - Expansion of their IP Portfolio through a patent - Exploitation of the produced results, mainly in the form of royalties or licensing of the technologies that will be developed

The Project

  • WP1: Design and Construction of sensors

  • WP2: Development / Evaluation of Reagents and Comparative Analysis Methods

    • WP3: Implementation of a Metering System

    • WP4: Development and Preliminary Evaluation of the sensors

      • WP5: Evaluation of the FOODSENS microsystem by the food industry

        In WP1 - Design and Construction of Photon Tiles, initially, single-analyzer tiles will be designed and manufactured, which will be used for the development of signal processing software, as well as the preliminary evaluation of the system in terms of sensitivity and dynamic range per parameter/target. The dual analyzer tiles will then be manufactured for use in WP4 and WP5.

        WP2 - Development / Evaluation of Reagents and Comparative Analysis Methods will focus on the development of enzyme-linked immunosorbent assay methods for target analysis, as well as the development of sample processing protocols. At the same time, the selection and development of comparative methods (LC-MS / MS and microbiological) will be carried out through which the laboratory evaluation of the FOODSENS system will be performed, while studies of the presence of pathogenic microorganisms in dairy products will be carried out with emphasis on phenotypic / biochemical characterization of strains of B. cereus and L. monocytogenes.

        WP3 - Has as its main objective the design and implementation of the FOODSENS device, both in terms of hardware and software. Emphasis will be placed on the development of a special application for a user-friendly portable device that will not require specialized personnel for its operation. A total of three provisions are envisaged, one for the preliminary evaluation studies and the other two for the full evaluation.

        WP4 - Development and Preliminary Evaluation of Biosystems will deal with the chemical and biological activation of the sensors, the development of immunosorbents for individual analyzers / microorganisms, as well as the development and preliminary evaluation of immunosensor for synagogue / microsensor. Important WP4 objectives are the stability of immobilized biomolecules, finding common assay conditions for aflatoxin M1 and bovine casein, for both bacteria to facilitate the implementation of concurrent detection protocols with dual biosets, and the determination of the analytical characteristics of the methods (detection and quantification limits, linear response regions, repeatability within and between assays, accuracy).

        These 4 WPs that will constantly interact with each other will lead synergistically to WP5 - Evaluation of the FOODSENS microsystem by the food industry, i.e. the pilot implementation of the FOODSENS rapid analyzer / microorganism detector in real conditions. An important part of the WP5, and one of the main objectives of the project is the preparation of a business plan and the definition of a strategy for the commercial use of the FOODSENS device, while a special sub-action will undertake the actions of promotion and dissemination of results in both academic and the business community. Finally, for the smooth and successful completion of the project, the potential risks and

        Finally, for the smooth and successful completion of the project, the potential risks and the necessary alternatives have been foreseen. Thus, in case the limits and detection times targeted for the analysts cannot be achieved without the use of probes, their sample pre-concentration systems and signal amplification systems will be used without incurring the cost of analysis (~0.05 €/sample). Regarding the risk of patenting similar technology by other groups, a thorough search for relevant patents has already been carried out worldwide and it was found that until the date of submission of the proposal there is no relevant patent other than those held by ENM and EAP. However, the search for relevant diplomas will continue uninterrupted until the completion of the project.