OBJECTIVES

The overall objective of this project is to develop efficient intervention strategies, enhanced process control criteria, monitoring sampling schemes, and an easy-to-use food safety decision-support IT tool for artisanal food producers, aiming to the reduction and control of foodborne pathogens such as Listeria monocytogenes, Salmonella spp., Shiga toxin-producing Escherichia coli (STEC) and Staphylococcus aureus in selected traditional food products produced in Mediterranean regions of Portugal, Spain, Italy, France, Greece, Morocco, Tunisia and Algeria. The artisanal foods are 15 fermented foods of meat or dairy origin with unique technological and sensory characteristics, chosen for research because they may be potential vehicles for the transmission of foodborne diseases due to inadequate or antiquated manufacturing practices or final products’ microbial instability. These artisanal foods may pose health risks to consumers as they are often produced with variable, mostly non-standardised productive processes, undergo short fermentation and, in some cases, are directly consumed without any cooking. The successful development of the proposed project will be achieved through multinational and inter-disciplinary research activities divided into the following six specific objectives:

1. Determination of the origin and routes of contamination with foodborne pathogens in the artisanal food chains to identify the manufacturing faults/risk factors favouring the growth of pathogens; which will be carried out through the conduction of manufacture surveys, and physicochemical and microbiological surveys of raw materials, mid-products and end-products sampled during actual artisanal elaboration;
2. Assessment and development of biopreservation strategies based on functional starter cultures and natural plant-based antimicrobial extracts to control pathogens and extend shelf-life of each of the artisanal foods;
3. Conduction of a series of fate studies of pathogens inoculated in prototype artisanal foods in order to understand the pathogens’ viability during fermentation, ripening and storage under traditional manufacture process and enhanced manufacture variants (i.e., use of biopreservatives and alternative process variables);
4. Development of dynamic predictive microbiology models that mathematically describe the growth, survival and inactivation of pathogens as affected by the intrinsic properties of the product (i.e., water activity, pH, and lactic acid concentration) and the enhanced manufacture variants;
5. Delineation of intervention strategies, optimisation of process variables, norms/standards and design of monitoring sampling schemes for the control of pathogens during the artisanal food processing, all of which will be derived from process risk models built upon the actual artisanal production processes (gained from (1)), alternative production processes (from (3)) and the predictive models of pathogens (developed in (4));
6. Construction of a prototype safety decision-support IT framework for the artisanal producers, intended to serve as a quality and safety self-evaluation tool for assessing the lethality of their (traditional and biopreservation-based) manufacturing processes towards pathogens, predicting contamination levels in final products, designing quality monitoring tools, such as sampling schemes and control charts, upon current and target safety levels, suggesting possible corrective actions, with the notebook capability of assessing quality improvements in time and accessing traceability information.

WORK PACKAGES

WP1: Management and coordination

The main purpose of WP1 is the smooth development of the ArtiSaneFood project, which will be achieved by attending to the five following actions:

1. To coordinate and supervise the project’s research and innovation activities according to the outlined work plan, ensuring the execution of corrective measures or contingency plans if critical risks arise, and monitoring both quality and timing of the project deliverables;
2. To implement mechanisms to ensure exchange of knowledge between partners and harmonisation of methods (i.e, experimental designs, laboratorial methods, data analysis and script coding), since the ArtiSaneFood’s concept relies on partners of distinct expertise/backgrounds applying comprehensive approaches in their own artisanal foods;
3. To coordinate the regular update and implementation of the Plan for Exploitation and Dissemination of Results (PEDR) and the proper management of knowledge and IP;
4. To ensure that partner PIs maintain regular communication with their local food producers; and
5. To carry out the administrative, legal and financial management and reporting of the project; and manage contacts with the EU Commission and establish effective internal and external communication.

Description of work

The main work to be performed in WP1 is related to the coordination of research and innovation activities; the planning of knowledge-exchange and harmonised protocols for laboratorial and data analysis to be assembled and adopted by the partners, the administrative and financial management; the organisation towards timely production of deliverables and reporting; and the resolution of eventual conflicts. In addition, liaison with EC and compliance with procedures described in the Grant and Consortium Agreements will be ensured through participation in EC project reviews and regular internal review of periodic reports.

WP2: Tracking surveys in the artisanal food production

The main objective of WP2 is to collect microbiological and physicochemical data on artisanal products, their raw materials as well as production environments in order to elucidate routes and patterns of foodborne pathogen contamination; characterise between-batch variability in pathogens’ levels; and identify manufacture faults and risk factors contributing to growth/survival of pathogens. The specific WP objectives are the following:

1. To map the target artisanal food chains.
2. To identify ingredients, processes and storage conditions with impact on the safety of the artisanal foods, and to characterise microbial within-batch and between-batch variability for the design of quality monitoring tools (WP7)
3. To sequence the pathogens isolated from the artisanal foods, in order to understand contamination patterns and aid in the preparation of strains for inoculation in the fate studies of pathogens (WP5).
4. To identify critical process variables that are risk factors for the development of pathogens in the artisanal foods, for their subsequent assessment in the fate studies of pathogens (WP5) and addition as parameters in the decision support tool (WP8).

WP3: Biopreservation by lactic acid cultures

The objective of WP3 is the selection of biopreservation strategies based on commercial starter cultures and/or indigenous lactic acid bacteria with antagonistic action against pathogens and good technological properties, whose efficiency will be tested mainly in-vitro, and preliminarily in-situ with the purpose of keeping the sensory attributes of the traditional formulation.

Description of work

While artisanal producers can keep good hygiene practices in the production of fermented foods, the end-products may still present an unsatisfactory microbiological quality and very short shelf-life if the fermentation process was not effectively regulated to provide microbial stability and safety. The use of lactic acid bacteria (LAB), with their known mechanisms of acidyfing the medium and producing bacteriocins, constitutes an interesting bio-preservation strategy to enhance the microbial stability of the fermented foods. Thus, the goals of WP3 are: (i) to evaluate the bacteriocinogenic potential of commercial starter cultures and/or indigenous LAB isolated from the artisanal food products; (ii) to select bacteriocin-producing LAB strains for the control of pathogenic bacteria, mainly L. monocytogenes, Salmonella spp., STEC, and/or S. aureus, which should also possess appropriate technological properties; and (iii) to develop a tailor-made or functional starter culture for some of the traditional foods that are currently not formulated with starter culture. Several bacteriocins are known to display synergistic effects when used in combination with other antimicrobial agents, including natural phenolic compounds as well as other antimicrobial proteins. Nonetheless, the combined effect While artisanal producers can keep good hygiene practices in the production of fermented foods, the end-products may still present an unsatisfactory microbiological quality and very short shelf-life if the fermentation process was not effectively regulated to provide microbial stability and safety. The use of lactic acid bacteria (LAB), with their known mechanisms of acidyfing the medium and producing bacteriocins, constitutes an interesting bio-preservation strategy to enhance the microbial stability of the fermented foods. Thus, the goals of WP3 are: (i) to evaluate the bacteriocinogenic potential of commercial starter cultures and/or indigenous LAB isolated from the artisanal food products; (ii) to select bacteriocin-producing LAB strains for the control of pathogenic bacteria, mainly L. monocytogenes, Salmonella spp., STEC, and/or S. aureus, which should also possess appropriate technological properties; and (iii) to develop a tailor-made or functional starter culture for some of the traditional foods that are currently not formulated with starter culture. Several bacteriocins are known to display synergistic effects when used in combination with other antimicrobial agents, including natural phenolic compounds as well as other antimicrobial proteins. Nonetheless, the combined effect of bacteriocins with potential plant-derived antimicrobials (assessed in WP4) will be also investigated in WP3 to avoid the development of resistant strains.of bacteriocins with potential plant-derived antimicrobials (assessed in WP4) will be also investigated in WP3 to avoid the development of resistant strains.

WP4: Biopreservation by natural extracts

The objectives of WP4 are: (i) to screen the antimicrobial activity of various plant-derived antimicrobial extracts; (ii) to sub-select antimicrobials that are both effective against pathogens and compatible with the sensorial attributes of the product (to be tested in-situ - in the target products); and (iii) to determine the in-situ effective doses of the selected antimicrobials and optimise their application in the artisanal foods by different methods, including incorporation, smearing (on cheese surface) and active antimicrobial packaging.

Description of work

WP4 is critical for selecting and evaluating in-vitro and then in-situ potential plant-derived antimicrobials for application in the target artisanal foods. Due to the strong flavouring character of the plant antimicrobials, it is imperative that only agents organoleptically compatible with the present products be targeted and, also, that the minimum required concentrations to produce the antimicrobial effect in the foods be investigated. To do that, we need first to determine their activity in-vitro; and then assess higher fold-levels of their in-vitro effective concentrations at in-situ level. WP4 will also assess different modes of application of the selected antimicrobials, in order to maximise their efficiency without compromising the sensory properties of the food (Kapetanakou and Skandamis, 2016). Such methods include spraying, nano-emulsions, nano or micro-encapsulations in aqueous capsules and gradual (or controlled) release, via active antimicrobial packaging applications (Donsi et al. 2014; Shah et al. 2012).

WP5: Fate studies of pathogens in artisanal foods

The objective of WP5 is to acquire, under laboratory-controlled conditions, dynamic data on the growth/survival of pathogens throughout the processing of artisanal fermented foods manufactured under the traditional variant (control) and alternative variants (i.e., enhanced process variables and biopreservation methods as intervention strategies). To achieve this goal, a series of survival studies of the most representative pathogenic species inoculated in prototype artisanal foods will be conducted in order to understand the pathogens’ viability during fermentation, ripening and storage conditions.

Description of work

To mathematically characterise the kinetics of pathogens in the artisanal foods during manufacture, a series of fate studies must be first conducted to assess the effects of fermentation/maturation and smoking/ripening conditions on the population of pathogens under the different manufacture variants (i.e., traditional and those enhanced by the intervention strategies). Pathogens are to be inoculated in the raw ingredients during mixing (i.e., sausage batter and milk) and followed throughout the complete lab-scale processing mimicking as much as possible the actual process. The outcome of this WP is to produce reliable microbial kinetic data in order to understand how pathogens in the artisanal foods are affected by (i) the enhanced or stabilised process variables – identified by analysing the food chain data from WP2 – and (ii) the biopreservation methods previously assayed in vitro and in-situ in WP3 and WP4.

WP6: Dynamic modelling and process safety optimisation

1. To characterise the growth and survival of selected pathogens in 15 artisanal fermented food products by applying dynamic predictive microbiology modelling to extract microbial kinetic parameters from the data generated in the fate studies (WP5).
2. To ensure the safety of artisanal processes by using the dynamic predictive models for the optimisation or tuning of those “critical” process variables that had been earlier (in WP2) determined as risk factors leading to high counts of pathogens in the end products.
3. To advance the state-of-the-art of Predictive Microbiology by developing innovative approaches for both dynamic predictive modelling and safety optimisation of fermentation processes.

Description of work

In WP6, predictive microbiology models will be constructed to mathematically describe the viability of pathogens in the artisanal fermented foods, as affected by the intrinsic properties of the product (i.e., Aw, pH and/or lactic acid concentration) and their manufacture characteristics (i.e., addition of commercial/functional starter cultures and/or plant-based antimicrobials, and enhanced process variables). By fitting accurate dynamic models to the data produced in the fate studies, not only the kinetic parameters of pathogens in 15 artisanal foods will be for the first time characterised, but also relevant process variables will be optimised or fine-tuned. WP6 subdivides into two Tasks.

WP7: Intervention strategies in the artisanal food production

Using the outputs generated in WP2, WP5 and WP6, WP7 (i) will build process risk models of pathogens covering the entire manufacturing process and shelf-life, in order to rank the single and combined intervention strategies (i.e., enhanced/adjusted process variables and biopreservation approaches) by mitigation efficacy; and (ii) will derive thereof science-based norms/standards and quality monitoring tools.

Description of work

Process risk models will be built for foodborne pathogens related to each of the 15 traditional fermented products to rank single and combined intervention strategies mitigating the foodborne disease risk. The models will also be used to derive risk-based standards and quality monitoring tools. Since each of the partners focuses on at least one artisanal food, the development of this WP will be common to all the Mediterranean partners. USDA-ARS will participate in this WP, not by programming process risk models, but by providing advice and technical expertise for a harmonised model development among partners (i.e., the 15 food products).

WP8: Safety decision-support tool for Mediterranean artisanal food producers

WP8 aims to develop a safety decision-support IT tool, assembling all project’s outputs, to enable artisanal producers to assess the safety of their traditional and biopreservation-based manufacturing processes, and generate sampling schemes and control charts upon their current and target safety levels.

Description of work

Built upon WP2, WP6 and WP7, a decision-support tool will be proposed for artisanal producers to evaluate the safety of their foods, under both traditional and new manufacturing processes, and to be able to derive risk-based process monitoring tools such as in-house sampling plans and control charts.

WP9: Dissemination, communication and education

The objective of WP9 are: (i) to strengthen the multidisciplinary network of consortium members with varied scientific and technical skills by means of an efficient information-flow management, interactive working mechanisms across the WPs, and technical webinars/seminars for knowledge transfer between partners; (ii) to promote the close involvement of artisanal producers in the ArtiSaneFood project, particularly at the points of collecting food chain data, testing intervention strategies and delineating the capabilities of the decision-support IT tool; (iii) to enlarge the state-of-the-art in pillar scientific areas of the ArtiSaneFood project; (iv) to promote and divulgate the ArtiSaneFood’s safety decision-support tool before other Mediterranean artisanal producers to stimulate its use; before European and national policy-makers to support tool’s deployment and sustainability; and before the scientific community to encourage further research and cooperation for the incorporation of other artisanal foods into this IT tool; (v) to increase the science uptake of all stakeholders, with emphasis to reach the artisanal food producers and disadvantaged or poorly educated people from the Meditarreanean; (vi) to promote the career development of young researchers and overall capacity building through mobility between partner instituitions; and (vii) to exploit the project’s results for stimulating the purchase of Mediterranean artisanal foods by reassuring the consumer in terms of tradition, quality, safety and benefits to the region.