Periodic Reporting for period 2 - MOLOKO (Multiplex phOtonic sensor for pLasmonic-based Online detection of contaminants in milK) Reporting period: 2019-07-01 to 2022-03-31 Summary of the context and overall objectives of the project The objective of MOLOKO project is the real-time monitoring of up to 10 food safety and quality indicators including antibiotics, toxins and quality parameters in milk by means of a self-managing and automatic integrated photonic sensor.By delivering a detailed reading in about 10 minutes, the sensor can look for any antibiotics ingested by the cows that have been transmitted to the milk. Using the collected information, companies can prevent contaminants to enter the food chain. Compared to the standard tests that take days to perform across dairy farms today, this new integrated optical sensor will be the easiest way to determine the presence of milk components (such as casein proteins and lactoferrin) that are quality parameters for milk and dairy products.Moreover, innovative assays were developed for detecting species’ milk fraud and A2-milk fraud with high precision, and Staphylococcus Enterotoxin A (SEA) according to target MRL with multiple detection approaches.The innovative technology is based on organic photonics, nanoplasmonics, biodiagnostics with immunoassays and microfluidics on a platform fit-for-use in milk production facilities.The MOLOKO project aims at developing and in-field demostrating a portable in-cloud sensor for the on-site monitoring of milk quality throughout the entire value chain, which can be also directly integrated in milking machine thus allowing early-alert assessment. Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far The major scientific/technical outputs achieved by the end of the project (M51) are:- Optimization of the surface biofunctionalization and assembly protocol of the Optoelectronic Module- Calibration and self-testing functions of the MOLOKO sensor- Implementation of a predictive model for the analysis of the raw data generated by the sensor- Demonstration of the multiplexing of the nanoplasmonic sensing surfaces- Demonstration of the detection of both low-molecular-weight (LMW) and high-molecular-weight (HMW) targets by using the MOLOKO manual prototype- Development of a robust authenticating multiplex method, capable of detecting species’ milk fraud and A2-milk fraud with high precision- Identification of recombinant antibodies (Fab-fragments) binding different epitopes of Staphylococcus Enterotoxin A (SEA)- Validation o the multiplex functionality of the biofunctionalized sensing surface designed and fabricated for the use in the MOLOKO sensor- Assembly of the prototype of the automatic sensor and check for working properly in real-setting applications (i.e. in milking parlors)- Integration of the MOLOKO automatic sensor into milking machines and process by the realization of an optimized prototype of Milk Sampler at Milkline- Internal comparison of MOLOKO sensor with benchmark SPR-method- Measurements of milk spiked and raw samples in MOLOKO sensor- Quantitative evaluation by Parmalat that Lactoferrin monitoring by the MOLOKO sensor could be an effective parameter (biomarker) to early detect animals that are healthy and are not affected by mastitis- Integration of MOLOKO sensor into HACCP-like multi-sensor semi-automated system (BEST platform)The major outputs relating to Exploitation and Innovation management achieved by the end of the project (M51) are:- Realization of an IPR report: the major output is the recommendation of considering licensing methods for the IPR and bestowing certain rights to the licensee for future commercial agreements- Realization of a Business Plan where the estimated costs of manufacture of the main consumable sensor cartridge are reported along with overall business expenses and different operating scenarios. The reusability of the sensor cartridge has a large effect on profit margins: if cartridge reusability higher than 20 a stable business with consistent profit through repeat consumable business is evident. - The individual partner results were summarised and concise 5 commercialisation paths for the partners’ results with direct commercialisation potential- Exploitation activities are underway with 2 companies interested in manufacturing the sensor prototype- New funding opportunities for bringing forward the overall MOLOKO Sensor Platform have been identified, in particular EIC Accelerator programme is considered to be the most relevant to the continuation of MOLOKO-generated innovations to be pursued by PLASMORE specifically as a SME Considering the dissemination and communication activities, different actions have been performed for the whole duration of the project with the aim of involving different target groups. The participated events, for example, were addressed to the end-users’ sectors, with the aim of involving different target groups. A total number of 9 open access scientific publication have been published and 3 are under submission. Dissemination impact has increased in the second part of the project thanks to the organization of dissemination workshops that maximized the project visibility and gave important feedbacks on the results achieved and next steps to improve the MOLOKO sensor. More that 70 events/actions have been put in place for disseminating the results of the project Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far) Strict monitoring of raw material and semi-finished products quality parameters are emerging as an invaluable tool for improving the general efficiency of the dairy chain and, particularly, in abating production losses and enhancing the nutritional and organolectic constancy and quality of the finished product. Quality and safety control in the dairy industry may generate a large number of analyses with significant associated costs. On the other side, consumers demand for high quality products, competitiveness, and efforts to reduce food waste and losses motivate innovation. The development and application of fast, sensitive and cost-effective analytic systems for residues, contaminants, pathogens and quality parameters in milk could aid the industry in the reduction of overheads, find new uses in dairy farming and production precision management and unlock new markets.The development and application of fast, sensitive and cost effective analytic systems for pathogen detection in milk along the production and supply chain will aid the industry to increase the efficiency of the process monitoring and control, with consequent reduction of overheads and good waste, find new uses in dairy farming and production management and unlock new markets.MOLOKO technology introduces a disruptive advance in realization of high-performing low-cost in-field monitoring tools aiming at (i) changing the de-facto milk and dairy supply-chain processes and the personal and institutional approaches to food contamination detections; and in general (ii) increasing food safety, public health and security of food supply-chainMOLOKO proposes proposes ground-breaking approaches in addressing the technological issues that limit the unrevealing of the full potentiality of biosensors in bringing the laboratory-quality analysis to on-line process monitoring and control.The challenges associated with the development of the innovative MOLOKO opticla biosensors are:- the increase of the overall method sensitivity and instrument miniaturization- the increase reliability and specificity- the feedback to the user-driven specific needs and new applications- the definition of the effective standardization protocol from farm to fork in the diary supply chain MOLOKO logo in the MOLOKO brochure
