Final Report Summary - QUAFETY (COMPREHENSIVE APPROACH TO ENHANCE QUALITY AND SAFETY OF READY TO EAT FRESH PRODUCTS)
Executive Summary:
The Project QUAFETY (Comprehensive Approach to Enhance Quality and Safety of Ready to eat Fresh Products) was aimed to improve quality and safety of Ready-to-eat (RTE) fresh produce throughout the whole chain by developing new predictive and probabilistic models and decision-making tools, by exploring rapid and non-destructive methods for quality evaluation and prediction, and by experimenting novel technologies, in order to quantify and manage spoilage and pathogen microorganisms, minimize risks to consumers, and preserve quality. The participants were 14, from 7 Countries and included 6 SMEs, 2 public research Institutes and 6 Universities, assorted in terms of scientific and technological expertise. The presence of SMEs ensured the exploitation of the results directly and indirectly.
As a response to the industry needs and in relation to the state of the art, within the project QUAFETY the following results were obtained:
- reliable and fast detection diagnostic kit based on key molecular probes for L. monocytogenes and E. coli O157:H7 and O104:H4, and on specific antibodies against the senescence associated proteins and against the human pathogen bacteria to detect stress conditions and presence of contaminants;
- rapid non destructive tools for process control which were aimed to better estimate shelf-life of fresh-cut products and to link the deterioration of external quality indexes to nutritional attributes, to identify nutritional, functional, and quality markers including volatiles to be monitored during process to reduce the impact of processing on final quality;
- decisions supporting tools to allow industry operators to understand and implement process innovations aimed to improve safety and quality, and particularly to select an “ideal melon” for processing, to predict barrier properties of polymeric films and the oxygen evolution in MA packaging, to estimate the respiration rate of fresh-cut packaged products and the impact of temperature and cutting on respiration kinetics, to predict bacterial lag time at various temperatures and finally to evaluate innovations starting from QUAFETY results which were included in an excel spreadsheet based software to support innovation investments;
- innovative processes and technologies were developed to improve quality and safety of the final products starting from raw material quality from pre-harvest (management of nutrient, varietal selection) to postharvest (passive refrigeration system, developing an automatic hands-off machine for melons) and processing (decontamination systems for water, products and industry plants, use of physical treatments and innovative packaging solutions);
- evaluation of the impact on markets and profitability for the industry considering consumer response to safety and quality attributes deriving from the potential application of QUAFETY innovation;
- development of a more effective and efficient Quality/Safety Management System for RTE fresh produce industry.;
The results of this project contribute to provide scientific evidences on factors affecting quality and safety of fresh-cut products with the aim of ensuring high quality standard throughout the chain. These results may be a reference point to mass media, and eventually to EC and other health authorities (also for campaigns for healthy nutrition), which will contribute to increase consumer awareness on quality and safety of fresh-cut products. Finally the level of innovation reached with the project will strengthen industry competitiveness.
Project Context and Objectives:
An important category of RTE food is represented by fresh-cut fruit and vegetables (F&V). The fresh-cut sector may be the smallest and most recent F&V segment in Europe, but it is clearly the fastest growing market. Fresh-cut salads are the favourite RTE fresh produce of European consumers, with more than 50% of the market volume, while fresh-cut fruits account for more than 10% of the share. These products meet new consumer's demands for healthy and easy to prepare food. However several outbreaks of food-borne diseases linked to the consumption of RTE produce have been recorded in past years.
One important thing related to safety of RTE fresh produce is that the preparation process does not include a “killing step”, or a treatment which reduce the final microbial count to value close to zero. This indicates that fresh-cut vegetables sold in the market may represent an underestimated public health risk due to the possible presence of pathogenic bacteria. Another peculiarity of RTE fresh F&V is that the food is constituted by living cell tissue, rapidly metabolizing, especially when peeled and cut in portions for higher convenience. Quality attributes (i.e. appearance, texture, flavour, and nutritional value) degrades very fast and shelf life is often a matter of days. For all these reasons the need of improvements in terms of safety and quality of fresh-cut produce is of paramount importance.
A number of priorities are on top of the list which may have a direct influence on RTE food consumers. SAFETY of course, not only in relation to microbial risks, but also related to other possible chemical contaminations, both from anthropogenic activities or naturally occurring. And QUALITY, especially considering expectations from consumers who pay a relatively high price in order to purchase a high-convenience fresh fruit or vegetable.
As a response to the industry needs and in relation to the state of the art, QUAFETY aims to improve safety and quality of RTE fresh produce throughout the whole chain, addressing the following objectives:
- to provide the industry with diagnostic kits for the evaluation of microbial contamination and shelf-life;
- to provide useful tools for process control to the fresh-cut industry based on non-destructive and rapid measurements, aimed to enhance the final quality of the product;
- to provide the industry operators with decisions supporting tools in very critical points of the fresh-cut processing chain from raw genotype selection to economic strategic planning- to improve, develop and implement process innovations aimed to improve safety and quality;
- to consider consumers’ response to safety and quality attributes deriving from the novel applications in order to evaluate the impact on markets and profitability for the industry;
- to design and implement a more effective and efficient Quality/Safety Management System for RTE fresh produce industry;
- to disseminate in a modern and efficient way all the innovative products of the present proposal.
The objectives of the proposal were reached through the realization of 9 Work Packages (WP) strongly interconnected, as shown in the conceptual diagram (Fig. 1). WP1 developed diagnostic kits, WP2 was dedicated to process control aids, WP3 developed decision support tools, WP4 investigated innovative processes, whereas technological innovations went through implementation and demonstration in WP5 and economic evaluation in WP6. All results represented valuable information for a more efficient management system for quality and safety (WP7) disseminated to potential users in WP8; WP9 dealt with the management of the consortium.
The objective of WP1 was to develop diagnostic kits for the detection of microbial contamination and the quality losses, reached through 4 tasks carried out from 3 different partners. The aim of Tasks 1.1 and 1.2 was to develop fast and reliable detection methods of L. monocytogenes and E. coli O157:H7, also through the individuation of key molecular probes able to detect genes involved in pathogenicity. The third task (Task 1.3) reported the research activities focused on the development of antibodies against the pathogens involved in major outbreaks of foodborne diseases in order to detect the presence of contaminants, while Task 1.4 was related to the individuation of molecular markers linked to physiological degradation of the produce, such as lipid peroxidation, ascorbic acid and chlorophyll degradation. During the second period, according to the recommendations received by the EC reviewers, researchers involved in this WP extended the validation of PCR methods developed during the first period for L. monocytogenes and E. coli O157:H7 and qPCR method for E. coli O104:H4, on non-artificially-inoculated samples, and also evaluated the prevalence of L. monocytogenes and E. coli O157:H7 in melons.
The objective of WP2 was to provide to the fresh-cut industry useful tools for process control based on non-destructive and rapid measurements or evaluation, aimed to enhance the final quality of the product, and it was organized into four tasks. Task 2.1 was aimed to study degradation kinetics of quality attributes for fresh-cut melons and for rocket leaves, identifying relationships between the degradation of external and internal quality attributes and validating the mathematical model developed. Task 2.2 was mainly focused on the determination of relevant nutritional and functional quality markers for fresh-cut melon, strawberries and ready-to-eat rocket leaves and on the optimization, selection and implementation of the most efficient techniques to assess these representative markers (chemical and/or biochemical markers). Moreover it was intended to determine throughout storage the effect of different processing and storage conditions on the quality markers, namely cut sizes, modified atmosphere packaging and storage temperature. The objective of Task 2.3 was to study the effect of different processing steps on quality markers (determined in Task 2.2) on functional and nutritional properties. Task 2.4 was aimed to study the volatile organic compound (VOC) profiles associated with cantaloupe melon and rocket salad, and to assess profiles of samples from other partners, to individuate optimal pre-and postharvest handling conditions. After the midterm report, the EC reviewers requested to extend the VOCs study also to strawberries as non-destructive markers for quality and safety. Therefore, a further objective was to obtain preliminary data for whole and halved strawberries using the TD-GC-MS TOF system.
The general objectives of WP3 were to provide tools for the industry operator to support decisions in very critical points of the fresh-cut processing chain from raw genotype selection to planning economic strategy for investments. The WP3 was articulated in 6 tasks, each one with a different objective. Task 3.1 was aimed to define morphological, physiological and biochemical parameters of an “ideal melon” most suited for fresh-cut processing. The objective of Task 3.2 was to determine the kinetics of respiration of fresh-cut fruit and vegetables as a function of oxygen partial pressure, and to evaluate the effect of temperature and cutting on these kinetics, developing a physiological model to estimate the potential benefits of MAP on melon and rocket leaves. Task 3.3 was related to the development of mathematical models to predict the barrier properties of polymeric films and the respiration rate of fresh-cut packaged products. The objective of Task 3.4 was to build a user-friendly decision-support tool for predicting oxygen dynamics in MA packages and to test its performance with fresh-cut melons. The main objective of Task 3.5 was to determine the single cell lag phase duration of undamaged and sub-lethally damaged (by heat and acid) pathogenic bacteria (Listeria. monocytogenes and Escherichia coli) and develop a model to predict lag time at various temperatures. Finally Task 3.6 wanted to provide a decision support tool enabling industry to better understand innovations produced within QUAFETY and select those coherent with their prioritization in terms of innovation needs and minimum requirements for the scaling-up.
WP4 was aimed to develop innovative processes to improve quality and safety of the final products, it was led by ARO and organized into 9 tasks.
The purpose of Task 4.1 was to obtain in several leafy vegetable species a raw material for fresh-cut processing with improved characteristics in terms of safety, nutritional quality and shelf-life, by strategically managing the nutrient solutions in different soilless cultivation systems, including the definition of innovative optimized growing protocols for several model species as lettuce, endive, escarole and rocket salad.
Task 4.2 was related to the implementation of chlorine-alternative method for water and product disinfection. The goal of Task 4.3 was to improve the efficacy of decontamination of raw melons intended for fresh-cut processing in order to ensure microbiological safety and to control spoilage microorganisms on ready-to-eat products. The objective of Task 4.4 was to analyze the main critical factors affecting the formation of Listeria monocytogenes biofilms in the fresh-cut industry and to find the efficient approaches to its control. The purpose of Task 4.5 was to improve efficacy and safety of fresh-cut melon and watermelon processing by substituting manual peeling and cutting operations by an automatic hands-off machine. Task 4.6 was aimed at testing a novel postharvest heat shock treatment as means for improving quality and safety of fresh-cut fruit and vegetables and extending their shelf life. The objective of Task 4.7 was to examine the efficacy of ultraviolet (UV) light for improving quality and safety of fresh cut products. Task 4.8 was aimed at addressing two types of packaging: (a) active and (b) intelligent. The active packaging was based on active antimicrobial compounds using two methods of their release to the product surface: (i) from the packaging through the headspace and (ii) via edible coating. Regarding the intelligent packaging, the work aimed developing a novel Time-Temperature Indicator of the product freshness. The general objective of Task 4.9 was to ensure the cold chain maintenance by applying a Passive Refrigeration System (PRS), transportable and stackable, to be used for fresh-cut products throughout the supply chain.
The WP5 was mainly aimed to demonstrate the efficacy of the developed technologies to ensure safety and quality of ready-to-eat fresh produce throughout the whole chain. The activities were articulated in four tasks, one for each of the previous WPS, Task 5.1 (Demonstration activities for diagnostic kits), Task 5.2 (demonstration of process control systems), Task 5.3 (Demonstration of decision support tools) and Task 5.4. (Demonstration of innovative processes). Therefore, demonstration was aimed to prove the technical viability of the new technologies, thus expecting to speed up the adoption of the developed innovative tools at the industrial level.
The WP6 was aimed to reach two main objectives. The first referred to the modelling and calculation of willingness to pay, based on consumer survey data and preferences analysis (Task 6.1). The second was related to the cost-benefit analysis, which took into consideration the results of willingness to pay, together with the results of a questionnaire filled by QUAFETY investigators (Tasks 6.2 and 6.3) in order to calculate cost information relative to deriving innovations and measure the level of profitability for each innovation produced within the project.
WP7 objectives were aimed to design tools to support the implementation of an effective and efficient Quality and Food Safety Management Systems in the fresh-cut produce sector. These tools included the development of a validated diagnostic instrument (questionnaire) to evaluate best practices in SMEs and the design of a Safety Assessment Plan (SAP), developed in the Task 7.1 to determine food safety along the production – processing chain. This SAP protocol was used during a pilot test, involving several SMEs within the Task 7.2 and finally the results of the questionnaire addressing technological and managerial elements critical to QMS/FSMSs and of the SAP were critically compared to assess the overall performance of SMEs and its potential improvement in the Task 7.3.
Finally one of the general objective of the project very important for all partners was related to the dissemination of the results obtained (WP8). The 7 tasks of WP8 were aimed to disseminate the project results to potential users (i.e. food industry, consumers, health authorities, mass media) during the project lifetime, through the project website (www.quafety.eu) organization and participation to events, papers and divulgative articles, and in particular through the capillary distribution of the online newsletters and websites of FreshPlaza (www.freshplaza.com one of the most visited websites dealing with postharvest technology and innovation, published in English, Spanish and Italian), currently used as an indispensable source of daily news among the professionals which operate in the fresh produce industry.
Project Results:
S&T results on diagnostic kits (WP1)
The aim of the research activities carried out in the WP1 were oriented on the identification of reliable and fast diagnostic kits able to detect the presence of human pathogen bacteria such as Escherichia coli and Listeria monocytogenes and assess the quality of fresh cut vegetables. In this WP were involved the University of Foggia (UNIFG), the Agricultural University of Athens (AUA) and University of Milano (UMIL).
UNIFG was involved in Task 1.1 for the identification of molecular markers to identify food borne pathogens such as Listeria monocytogens and E. coli O157:H7 in fresh cut vegetables. Firstly, a protocol for bacterial DNA extraction from vegetables samples, suitable for pathogen detection in PCR and Q-PCR assays was reported (deliverable D.1.1). Furthermore, protocol has been improved comparing several DNA extraction kits commercially available. A final DNA extraction kit was than chosen and used for all the experiments carried out. The occurrence of Listeria spp. and E. coli O157:H7 in minimally processed vegetables purchased from different retailers in Apulia region (Italy) was investigated, using both selective media, immunoassay tests and PCR detection tools. Moreover, MPN-QPCR methods for quantification of L. monocytogenes and E. coli O157:H7 from fresh cut vegetables and enrichment free methods for quantification of L. monocytogenes, E. coli O157:H7 and E. coli O104:H4 by Q-PCR were developed.
Primers were chosen among the ones available in literature in order to design QPCR assays that were highly sensitive and specific for purposes. A total of four QPCR assays were designed and tested for Listeria monocytogenes. Three of the primer pairs employed targeted virulence-related genes (iap, hlyA), while one set was used to amplify a sequence in the 16S rRNA gene. Two different chemistries, SYBR green (for 16S rRNA and hlyA(1) assays) and TaqMan (for hlyA(2) and iap), were used to set the QPCR assays. For E. coli O157:H7, primer sets targeting two different genes were tested during the study. Primers were selected from literature since are widely used in reference laboratories across Europe, for detection and enumeration of E. coli O157:H7. Both primer sets were coupled with fluorescently labelled probes in order to perform TaqMan assays. Target genes are rfbE, encoding antigen O157, and fliC H7, encoding flagellar antigen H7.
For E. coli O104:H4 primer sets and probes targeting genes wzxO104 and fliCH4 were tested during the study. These primers have been successfully employed for detection of E. coli O104:H4 during the large outbreak of 2011 in Germany. Both primer sets were coupled with fluorescently labelled probes in order to perform TaqMan assays.
For L. monocytogenes the MPN-Q-PCR method developed, can be used to shorten the MPN procedure from about 7 days to one day with a faster and reliable confirmation of L. monocytogenes in enriched samples (Fig. 2). The assay allows detection of L. monocytogenes down to 101 cells/g without enrichment and quantification of cells in concentration down to 102 cells/g after 6h enrichment.
For E. coli O157:H7, the MPN-Q-PCR method developed can be used to shorten the MPN procedure from 4 days to one day with a faster and reliable confirmation of E. coli O157:H7 in enriched samples. The assay is also suitable for rapid detection of pathogens as it allows detection of E. coli O157:H7 down to 101 cell/g after 2 hours of enrichment (See first period report). For E. coli O104:H4, the selected assays had proven to be highly specific and sensitive. Assays based on the oligonucleotides correctly identified E. coli O104:H4 in all of our tests while showing no signal in case of identified E. coli O157:H7 and non-STEC E. coli. The MPN-Q-PCR method developed was further confirmed in cooperation with the Istituto Zooprofilattico per la Puglia e Basilica - IZSPB (Foggia, Italy) on randomly selected fresh cut vegetables.
The aim of the research activities of the Task 1.2 performed by AUA were to assess the: (1) prevalence of L. monocytogenes and E. coli O157:H7 in ready to eat fresh products, (2) genotypic diversity of the respective isolates in terms of genotype, serotype and key gene content and (3) effect of storage conditions on the transcription rate of key genes at the ecosystem level.
Prevalence of L. monocytogenes was 7% in rocket, 6% in cucumbers, 6.6% in melon rinds, and 3.8% in strawberries. Prevalence of E. coli O157:H7 was 7% in rocket, 3% in cucumbers, 6.6% in melon surface and 3.8% in strawberries. Both pathogens were not detected in the edible part of the melon samples analyzed (prevalence 0%). Accurate estimation of prevalence could only be achieved by the parallel use of at least two chromogenic media; using only one chromogenic medium in fact leaded to over- or under- estimation of the prevalence depending on the medium. The ability to detect the pathogens in the enrichment broths through specific PCR and RT-qPCR was also examined concluding that plating chromogenic media with inhibitory agents against antagonistic microbiota cannot be reliably replaced by direct PCR or RT-qPCR in the enrichment broths.
Regarding the genotypic diversity of the isolates, all of them, even from the same sample or chromogenic media, were differentiated by genotypic profiling at the strain level. All isolates contained the necessary genes required for pathogenicity, i.e. iap, prfA, plcA, plcB, hly, actA, inlA, inlB, inlC and inlJ regarding L. monocytogenes and stx1, stx2, eae and ehxA regarding E. coli O157:H7, as detected by published specific-PCR protocols, with modifications due to possible small differences in the gene sequence.
Regarding the effect of storage temperature (4, 10 and 30 °C), product type (melon, rocket) and time (upon inoculation, 0.5 6.0 and 24 hours) on the transcription of key virulence genes of the pathogens, it was concluded that the transcription of hlyA, plcA, inlA, sigB, iap, plcB, inlB, inlC, inlJ, lmo2672 and lmo2470 at ecosystem level was indeed affected by the conditions under study. On the contrary, transcription of stx1, stx2, eae and ehxA were not affected by these factors.
UMIL was involved in the identification of molecular markers to identify microbial contaminants and to evaluate quality during the whole production chain. During the first year, the research group at UMIL in collaboration with UNIFG and AUA developed and optimized an ELISA method using specific antibodies against Escherichia coli and Listeria monocytogenes. Different buffers and strategies were tested in order to isolate the bacteria from inoculated fresh-cut vegetables. The protocol developed and optimised allowed the isolation and detection of the human pathogen bacteria in 22-24 hours. The method was applied on different fresh cut vegetables such as cucumber, melon and rocket. The validation of the method was carried out using samples collected on the market and during the demonstration period as suggested from the referees of the middle term project evaluation.
The isolation of putative molecular markers for quality evaluation was performed by transcriptome analysis of rocket plants exposed to different pre- and postharvest stresses. The sampling was performed after 24 h since the aim was to avoid the isolation of primary genes activation but those that might be really associated with quality losses. The transcriptome analysis yielded more than 30,000 transcripts and few of them were commonly expressed in all stresses. Since, the project was focused on fresh-cut the molecular markers were searched among those that were commonly upregulated in postharvest stresses. The identification of the putative genes as molecular markers was carried out considering their intensity of expression. The behavior of the most promising genes was studied during postharvest of rocket and melon. The putative molecular markers genes were correlated with physiological and biochemical parameters in order to be sure that they were associated with quality losses. The expression of these genes was studied in rocket and melon stored at 4 or 20 °C, simulating an optimized cold distribution chain and room temperature condition, which should speed up all the senescence processes. The genes that provided good results in both fresh-cut vegetables were used for antibodies production. In particular, the region of the genes that shown high conserved regions were chosen, because the idea was to identify molecular markers that can work for a wide range of vegetables. The antibodies were then used for the antibody array chip production. In particular, four antibodies were used for the antibody array production. The antibody array was tested as potential diagnostic tool for providing fast and reliable results on quality and safety of the fresh cut vegetable at any stage of the distribution chain. On the antibody array three antibodies against senescence-associated proteins, encoded from the genes identified from the transcriptome analysis, and one antibodies against E. coli were printed. The antibody array was tested on inoculated E. coli rocket samples and senescent rocket and melon samples. The results are very promising for bacteria detection and for quality assessment of fresh cut rocket. The use of this antibody array should give with one single analysis the overall quality of the fresh-cut produce.
S&T results on process control (WP2)
The aim of the activities carried out in WP2 was to provide useful tools for process control to the fresh-cut industry based on non-destructive and rapid measurements or evaluation, aimed to enhance the final quality of the product. In this WP partners UNIFG, Universidade Catolica Portuguesa (UCP), Cardiff University (CU), Nuvifruits and Markes International (Markes) were involved.
UNIFG worked on the prediction of the degradation of internal quality attributes of fresh-cut rocket salad and melons, starting from the degradation rate of external attributes (Task 2.1). This objective was reached in 3 steps: 1) obtaining the degradation kinetics of the most important quality attributes; 2) identifying relationships between the degradation of external and internal quality attributes; 3) validating the model. The first deliverable (2.6) consisted of two scales to objectively evaluate the appearance score of melon and rocket samples, which were also implemented in other partners’ activities such as CU/Markes, UCP/Nuvifruits and UMIL. The effect of temperature of storage on MA packaged products was modeled, by considering 3 storage temperatures. Data on melon and rocket showed that the “Weibullian-logistic” model was the best fit for the experimental data relating to the most important external and internal attributes (deliverable 2.7) compared to the traditional zero, first and second order kinetic models,. Moreover the model allowed the estimation of the shelf life both in isothermal and non-isothermal conditions. For the second step, degradation of external and internal quality attributes were related to each other by considering the storage time, t, as the only common variable (Fig. 3). A new mathematical equation able to estimate an internal quality attribute from an external one was obtained, also in this case in isothermal and in real condition simulations, highlighting its excellent ability to predict internal attributes (i.e. ascorbic acid content) through the easy measure of external indices such as appearance score, gas composition or color parameters.
Within Task 2.2 UCP focused at selection and implementation of efficient techniques to assess representative markers (chemical and/or biochemical markers) of nutritional and functional quality of ready-to-eat products with the final purpose of modelling and establishing the effect of different processing conditions on these relevant markers (Fig. 4). Fresh-cut melons and strawberries and ready-to-eat rocket leaves were fully characterized in terms of antioxidant capacity, and content of total antioxidant compounds (total phenolic content, total carotenoids, total anthocyanins). Qualitative and quantitative characterization of specific antioxidant compounds was also assessed to determine functional and nutritional quality. Specific conditions, namely package atmosphere and storage temperature were tested for ready-to-eat rocket leaves (rocket storage temperature experiments were carried out in collaboration also with CU/Markes) and fresh-cut melons, cut size for melon (in collaboration also with CU/Markes) and storage temperature for strawberries. All ready-to-eat products used in the experiments were supplied by NuviFruits and Estevão Luís Salvador.
Moreover, in Task 2.3 UCP together with Nuvifruits and Estevão Luís Salvador investigated the effects of the production process and storage on nutritional and phytochemical composition of melon, strawberries and rocket leaf products. Nutritional audits of processing lines of a fresh-cut fruit company and a ready-to-eat leafy vegetables enterprise were performed to generate recommendations for optimal preservation of phytochemicals of fruits and vegetables during processing through to the final product.
Audits were based on the enterprise’s definition of processing conditions (isolated or combined) of fruit and vegetables, their description in representative flowcharts, and identification of the process variables.
CU and Markes, performed an initial assessment of solid phase micro extraction (SPME) and thermal desorption, gas chromatography time-of-flight mass spectrometry (TD-GC-TOF-MS), and the latter was shown to be superior in robustness of collection method, sensitivity and lack of saturation (Task 2.4). This approach was therefore used to evaluate VOCs as a method for assessing effects of processing, storage time and temperature in melon and rocket salads, in collaboration with other QUAFETY partners. Following the mid-term review, the use of VOCs for assessing fresh cut strawberry quality was also evaluated.
In collaboration with UMIL and using scales developed by UNIFG, the complete VOC profile of fresh-cut melon cubes clearly differentiated samples stored at 4oC from those stored at 20oC, indicating that a breach in the cold chain would be detectable by this method. Several modules of compounds and selected single compounds were identified which look very promising for further development of VOC analysis for detection of storage temperature/ time evaluation. In collaboration with UCP, VOC profile analysis was shown to discriminate between different levels of melon processing. The discriminatory power was retained with the inclusion of only twelve compounds suggesting that the VOC analysis might be transferred to other platforms, and that by using TD-GC-TOF-MS, the analysis could be more targeted. VOCs could thus be used to assess changes in processing which may also be affecting nutritional value and or shelf-life.
In collaboration with UCP, the VOC profiles of rocket leaves were used to discriminate time x storage and showed good correlation to sensorial analyses and phytochemical content. VOC profiles also discriminated shorter term stress treatments of rocket leaves (in collaboration with UMIL) with a particularly strong effect of wounding, suggesting that VOCs may be of commercial use for assessing levels of mechanical damage during different processing pipelines.
Baseline data were obtained for fresh cut strawberries in line with recommendations at the mid-term review. A total of 63 compounds were detected of which 61 could be tentatively identified based on available databases, and the effect of storage time x temperature could be discriminated.
In collaboration with AUA, experiments on melon cubes inoculated with the human pathogen Listeria monocytogenes demonstrated that VOCs could be used to follow changes in levels of bacterial growth at different temperatures, and can detect levels of bacterial contamination. A 6 h period of incubation of the inoculated melon samples at 37oC increased the sensitivity of the VOC detection by 103 fold. This is still a short period of time compared to cultural or PCR based detection methods.
S&T results on decision support (WP3)
The aim of the research activities carried out in the WP3 was to provide tools for the industry operator to support decisions in very critical points of the fresh-cut processing chain from raw genotype selection to planning economic strategy for investments. In this WP partners Agricultural Research Organisation of Israel – The Volcani Centre (ARO), UCP, UNIFG, London Metropolitan University (LMET), Distretto Agroalimentare Regionale (DARe), Einat Food Industries Agricultural Cooperatives (EFI) and Nuvifruits were involved.
Within Task 3.1 ARO and EFI defined the criteria for choosing melon genotypes most suitable for fresh-cut processing and production of RTE melon products (an “ideal melon”). The performance of RTE products prepared from various melon genotypes was compared in storage trials. The study has shown that physiological behavior (climacteric vs. non-climacteric) affected product shelf life and off-flavour risk in response to packaging; rind surface (netted vs. smooth) affected the microbial attachment, decontamination efficacy and product safety; flesh color (orange flesh) reflected nutritional value associated with carotenoid phytonutrients; fruit size and shape affected the efficacy of manual and mechanical processing and waste amount; flesh texture and soluble solids level affecedt the product’s sensory quality. An inodorus-type melon cultivar Sorbeto was chosen as a model variety answering most criteria defined (Fig. 5). The performance of RTE Sorbeto melons was characterized in storage trials and further in demonstration activities (WP5) together with the industrial partner EFI and disseminated through WP8 activities. This genotype was used for developing RTE melon packaging (Task 3.4 WP3) and innovative mechanical processing (Task 4.5 WP4).
Moreover, in Task 3.4 ARO developed a method for optimizing modified-atmosphere (MA) packaging of RTE fruit in order to control their spoilage without a risk of oxygen depletion and off-flavour development. Microperforated packaging, with or without gas flushing (active MA) was used as a methodology to reach the aim. A user-friendly interactive decision-support tool predicting the in-package oxygen level as affected by package parameters (film type and thickness, perforation level, package dimensions, produce load, etc.) was built in order to adjust the packages for various commodities and conditions. The superior performance of the optimized MA packages was tested in storage trials with different melon cultivars (cantalupensis-type Raanan and inodorus-type Sorbeto) and further in demonstration activities (WP5) together with the industrial partner EFI. The results were disseminated through WP8 activities (Fig. 8).
UCP and NuviFruits performed the Task 3.2 related to the development of physiology-based models to assess the effect of oxygen partial pressure and storage temperature on ready-to-eat products respiration rate and quality parameters. Fresh-cut fruits and vegetables packaging trials were performed to allow modelling the degradation kinetics of quality changes and nutrient losses during post processing storage. Kinetic parameters found by linear regression for the orders 0, 1, and 2 allowed to select the most adequate kinetic order for the estimation of rocket leaves and fresh-cut melon quality changes under different modified atmospheres, and the goodness of fitting (Fig. 6). The best conditions found for the preservation of overall (physiological and biochemical) quality of ready-to eat rocket leaves and fresh-cut melons and strawberries were the initial package atmosphere of 5 % oxygen and the storage temperature of 5 ºC. Also, the functional description for respiration rate of ready-to-eat rocket leaves and fresh-cut melon, as function of oxygen partial pressure, was determined through O2 consumption levels monitored throughout refrigerated storage. A good fit of Michaelis-Menten model to experimental data was found and the kinetics of respiration as a function of O2 suggest that no significant reductions in respiration rate of fresh-cut cantaloupe can be achieved by lowering O2 levels. High R2 values for the Michaelis-Menten coefficients Vmax and Km were determined, although overestimation of O2 levels inside equilibrium modified atmosphere packages was common among fresh-cut produce tested. The results were obtained from WP2 parallel experimental trials and the results were disseminated through WP8 activities.
The Task 3.3 was performed by UNIFG in order to develop mathematical models to predict the barrier properties of polymeric films and the respiration rate of fresh-cut packaged products. The models can be used to provide information on the influence of the packaging characteristics (film thickness, presence of micro-perforations, etc.) on the respiration rate of fresh-cut packaged products (Fig. 7). This will make it possible to predict how to package a particular type of product only from the knowledge of its respiratory rate. For the first aim, monolayer, multilayer and micro-perforated films with different hole diameters as well as variable distance between them were taken into account, being different in terms of barrier properties. For the model validation a comparison between predictive and experimental data was carried out. In order to quantitatively determine the model prediction goodness, the mean relative deviation modulus was calculated. The model used to describe the packaged food respiration rate was obtained by writing the mass balance equation of gas in the package headspace of fresh-cut melon. The developed models are of great interest in packaging design of fresh-cut produce.
LMET studied the effects of mild (sublethal) heat treatment, growth at sub-optimal pH values (sublethal acid stress) and subsequent recovery temperature on the lag times of individual cells of Listeria monocytogenes and Escherichia coli (Task 3.5). It was found that, except for acid stressed L. monocytogenes, which showed a slightly short mean value for individual lag time at 10 oC compared with that at 22 oC, the mean and standard deviation (SD) of individual lag times of L. monocytogenes and E. coli increased when recovery temperature decreased. The mean and SD of individual lag times of L. monocytogenes increased after sublethal heat stress, while E. coli individual cells showed poor recovery at 10 and 15 oC after heat stress. Acid stress resulted in an increase of individual lag time for L. monocytogenes at 5 oC, but, surprisingly, in a decrease at 10 oC and no significant change at 22 oC, compared with undamaged cells. For E. coli, acid stress appeared to result in a decrease in individual lag at 10, 15 and 22 o C compared with the undamaged culture. The zero adjusted Gamma distribution was demonstrated to be the best fit to data for individual lag of both L. monocytogenes and E. coli. A quadratic polynomial relationship was developed to link the parameters of the zero adjusted Gamma distribution of individual lag with the recovery temperature. The performance of the model was validated by comparing the population lag of undamaged, heat stressed and acid stressed L.monocytogenes and E.coli in melon with values predicted by the model for the same environmental conditions. Except for the heat stressed model for L. monocytogenes, which provided a prediction for lag slightly comparable to the experimental population lag in melon, it was found that in most cases, the predicted population lag was significantly shorter than the observed lag in melon. Although the model “fails safe”, this underestimation of lag highlights the need for more conditions to be tested to improve performance of the model. In addition, L. monocytogenes cells subjected to combined acid and heat stress may pose a risk to consumers of fresh cut foods, such as melon, at the end of two weeks’ refrigerated shelf-life. Recovery of individual cells of E. coli after application of combined acid and heat stress was significantly affected by subsequent incubation temperature (Fig. 9).
In the Task 3.6 DARe conducted focus groups and in depth interviews among partners with the aim of identifying the innovations produced within the project, their impact on production and their requirements for the implementation. The outcome identified the key information for a direct communication between companies and QUAFETY research for a real understanding of the impact of the project on products or firms performance indicators. Having organized systematically the information DARe set up an excel spreadsheet based software providing a decisional tool to industry that will support innovation investments. The software has been, then, tested on a sample of 17 companies, which provided their feedbacks. The main outcome of the feedbacks concerns the possibility to better understand innovations and to furnish a more easy connection between innovations, their impacts on production, their requirements, and contact with researchers. In such a way, companies declared that with high probability will contact the researchers in order to have further information on the implementation in their production process (Fig. 10).
S&T results on innovative processes (WP4)
The aim of the research activities carried out in the WP4 was to develop and/or to evaluate the application of innovative processes to improve quality and safety of the final product. Looking at the whole farm-to-fork chain, the first step to consider is growing system and pre-harvest practices management. Partners involved in WP were ARO, UNIFG, the Instytut Ogrodnictwa (INSAD), EFI, and Agronomia Scarl (AGRONOMIA).
The purpose of the work performed by UNIFG in the Task 4.1 was to obtain raw material from several leafy vegetable species with improved characteristics in terms of safety, nutritional quality and shelf-life. The purpose was reached by strategically managing the nutrient solutions in different soilless cultivation systems. The research activity was focused on the definition of innovative optimized growing protocols for several model species including lettuce, endive, escarole and rocket salad (Fig. 11). The innovative protocols were based on soilless cultivation systems, considered the most intensive production method in today’s horticulture industry, using electrical conductivity levels of nutrient solutions from 1.5 to 4.5 dS m-1. Two cultivation systems were tested: a modified Ebb and Flow system and a Floating System (FS). Strategies aimed at the abatement of nitrate content in the leaves were also addressed. The following main results were obtained:
1. The best electrical conductivity of nutrient solution was 3.5 dS m-1. This level provides the best yield and quality performances giving leaves with low nitrate, high antioxidant compounds and high dry matter content as well as high shelf life;
2. The modified Ebb and Flow system appears more suitable with ‘lollo’ lettuce, while the Floating System performed the best with rocket salad, endive and escarole;
3. Nitrate content can be successfully lowered by substituting NS with water, but the strategy must be used no longer than the last 3 days before harvest;
4. The quality of soilless-grown rocket salad depended on season. The product grown at warmer conditions had better yield, visual quality, dry matter and vitamin C and keeping quality.
In order to reach the objectives of Task 4.2 AGRONOMIA tested the performance of water ultrafiltration system for pre-treatment, treatment and cleaning (disinfection) of water under the conditions of a commercial fresh-cut plant specialized on leafy vegetables processing. The ultrafiltration technology was best suited for the treatment of the process water, compared to quartz and metal mesh filters (Fig. 12). The ultrafiltration technique permitted to re-use the water in the process and to increase the volume of clean water used at the washing step. As an additional effect the system allowed to recover some of the water used for washing, and to put it back on circulation reducing the environmental impact of the whole system. In order to prevent trihalomethanes accumulation in ultrafiltration systems the hypochlorite was substituted with 2.5 ppm of chlorine dioxide sanitizer. The advantages of chlorine dioxide are its efficacy at low concentration, ease of use and lack of toxic byproduct residues. Combination of ultrafiltration with chlorine dioxide represents a significant alternative to chlorine in washing water sanitation of fresh-cut produce.
ARO (in cooperation with EFI) tested the efficacy of rind decontamination of melons intended for fresh-cut processing in order to ensure microbiological safety and to control spoilage microorganisms on ready-to-eat products (Task 4.3). A new method for thermal decontamination of melons and a prototype installation for realization of this method were designed and evaluated. The melon surface temperature reached 120 to 200 °C after 30 sec of the thermal decontamination. The treatment allowed the reduction of microbial population on the melon surface by 4.5 to 6 log CFU cm-2 while the efficacy of the standard hypochlorite treatment did not exceed 3.5 log CFU cm-2 (Fig. 13). After 30 s treatment, the heating was restricted to melon rind (less than 2 cm depth) and did not affect the flesh temperature. No negative effect on the fresh-cut product quality was observed. The optimal treatment duration was 20 to 30 seconds. The work has demonstrated the feasibility and efficacy of the proposed approach for decontamination of melons intended for fresh-cut processing.
Moreover, in Task 4.5 ARO improved the efficacy and safety of fresh-cut melon and watermelon processing by substituting manual peeling and cutting operations by an automatic hands-off machine. The study was designed to develop a technology and machinery able to mechanically cut and peel melons and watermelons in order to prepare fresh-cut, ready-to-eat products (Fig. 15). As part of this study several peeling alternatives ranging from laser-cut to high-pressure water using artificial vision systems, as utilized in sorting processes in the production industry, were compared. The study evaluated, analyzed and developed a number of special puncturing tools: fixed, rotating, solid and perforated. It was found that a round, disk shape slice as final product is achievable. The results of these trials have led to the design of the system performing peeling/cutting operations without a human hand touching the cut fruit. At this stage the machine requires a manual feed of the raw fruit. The main parts of the machinery have been built and tested successfully. The mechanically excised products were found to maintain the acceptable quality for more than a week when stored in sealed plastic packages in cooled environment.
Within Task 4.4 UNIFG analyzed the main critical factors affecting the formation of Listeria monocytogenes biofilms in the fresh-cut industry and to find the efficient approaches to its control (Fig. 14). The work strategy included three parts: (a) biofilm development in Listeria monocytogenes EGD-e and role of the agrA gene in biofilm formation; (b) biofilm formation in Listeria monocytogenes of vegetables origin; (c) inhibition and removal of established biofilms by L. monocytogenes using different oxidizing agents. The following significant results were obtained:
1. Temperature is a critical factor for biofilm formation by L. monocytogenes. Low temperatures should be maintained in the work environments in order to minimize the biofilm formation.
2. Listeria monocytogenes strains from vegetables origin are able to form biofilms. Biofilm formation is variable between strains.
3. Biofilms may be formed independently from the surfaces (glass, polystyrene and stainless steel) although hydrophobic materials should be preferred when compared to hydrophilic supports.
4. Commercial biocides may effectively remove biofilms formed by L. monocytogenes of vegetables origin. Sodium hypochlorite was effective when applied after the incubation of L. monocytogenes. In contrast, benzalkonium chloride and peroxyacetic acid were efficient only when applied before incubation of L. monocytogenes strains.
The activities carried out by INSAD in Task 4.6 were aimed to test a novel postharvest hot-water treatment as a mean for improving quality and safety of fresh-cut vegetables and extending their shelf life. During the reported period the research was carried out on Chinese cabbage, rocket leaves (salad and wild type) and melon (Galia and Cantaloupe type). The storability of Chinese cabbage cv. Bilko F1 was positively affected by brief (3–5 s) dipping in hot water at 53–55°C (Fig. 16). The hot water treatment delayed browning of cut cabbage leaves surface allowing to maintain better quality during storage. Sensory analysis performed after short term storage confirmed the superior sensory characteristics of cut Chinese cabbage dipped in hot water (53–55°C) as compared with non-treated control. Hot water treatments generally had no significant effect on chemical composition and microbial population of the fresh cut Chinese cabbage.
In contrast to Chinese cabbage, only limited quality improvement was reached with rocket and melons. With rocket, brief dipping (3–5 s) in hot water at temperature 50-55°C retarded the leaf yellowing during short term storage. However, the treatment was associated with the risk of heat injury.
The activities of Task 4.7 were performed by AGRONOMIA in cooperation with ARO to examine the efficacy of ultraviolet (UV) light for improving quality and safety of fresh cut products. UV treatment had a positive effect of reducing the microbial population on ready to use leafy vegetables. The greatest microbial inhibition was obtained using long time of exposure (more than 10 minutes, 8 kJ/m2) which caused a too high stress to the delicate rocket leaves. The smaller UV doses (2 or 4 kJ/m2) still had a positive effect of reducing the microbial population and at the same time exerted less phytoxic damage. With endive, the problem of the UV damage was far less acute than with rocket. The UV treatment resulted in certain increase of phenolic compounds content in rocket leaves and endive. The UV treatment proved itself an efficient instrument to inhibit the microbial proliferation during shelf-life of fresh-cut melons (Fig. 17). The treatment had no negative effects on the quality and nutritional value of fresh-cut melons and induced even a slight increase in the antioxidant activity. In the production of fresh-cut melons the UV treatment can be applied without hampering the processing chain when the fruit cubes are filled into open retail containers and before these containers are sealed.
In Task 4.8 , AUA, UNIFG and ARO worked together in order to develop active and intelligent packaging solutions for use with fresh-cut produce. The active packaging was based on active antimicrobial compounds using two methods of release to the product surface: (i) from the packaging through the hedspace and (ii) via edible coating. Regarding the intelligent packaging, the work aimed to develop a novel Time-Temperature Indicator of the product freshness. The following major results were achieved. (1) The vapors of lemongrass essential oil released by the packaging material affected the enterococci and yeast and mould populations depending on product, storage temperature and packaging atmosphere. The impact of the essential oil on product’s sensorial properties was pronounced in first 4-5 days of storage and faded as storage proceeded, due to the oil evaporation (Fig. 18). (2) T-gelatin films prepared by the Layer-by-Layer approach demonstrated good mechanical properties, higher permeability and effective microbial spoilage inhibition. The Layer-by-Layer (LbL) formulation was superior compared with monolayer formulations in preservation of fresh-cut melon texture and slightly reduced fruit weight loss. Both LbL and blended composite coatings did not obstruct fruit gas exchange and did not cause accumulation of off flavor volatiles. (3) The best combination of process variables corresponded to proper washing of the whole melon, dipping treatment of the cut pieces in hexanal, active coating with hexanal and packaging in OPP40 (Fig. 19). Under these conditions the shelf-life reached 16 days, compared to less than one week in the control. (4) A Time-Temperature Indicator based on physiological activity of lactic acid bacteria was developed and successfully tested with rocket leaves storage; its presentation on the packaging was proposed.
In Task 4.9 the work performed by AGRONOMIA as first step defined the specifications to fabricate a prototype of Passive Refrigeration System (PRS) container that could be used in the open field for collection of baby leaves or whole salads, for refrigerated transport, for short storage during processing, and for final distribution, maintaining throughout the chain the temperature between 0 and 4 °C. The seconf step was to test the PRS performance in the presence of product. The device confirmed its capacity to cool the product. The baby rocket leaves were found more prone to low temperature damage than the crisphead lettuce. The eutectic plate preparation temperature had a major effect on the efficacy of the PRS system (Fig. 20). With eutectic plate frozen at -15°C (completely frozen) the temperature in the PRS box dropped below the conventional forced-air transport conditions that might cause damage to rocket at pre-processing stage and to crisphead lettuce during shelf life. Eutectic plate chilled at 0°C (therefore not frozed) did not allow reaching appropriate temperatures, so the product encountered microbiological problems during storage, reducing the shelf-life in both commodities. Eutectic plate frozen at -4°C (partially frozen) allowed reaching appropriate temperatures in the PRS box comparable with the conventional forced-air refrigeration so that the products preserved similar or improved organoleptic and microbiological quality compared to traditional method. The limited quality improvement by PRS container compared with the traditional standard forced-air system was due to the brief time of transport (max 18-20 hours). Using PRS containers for longer distribution distances would maximize their benefits.
S&T results on demonstration activities (WP5)
Demonstration activities were aimed to prove the technical viability of the new technologies developed in the project, thus boosting at industrial level the adoption of new expertise and innovative tools able to ensure safety and quality of ready-to-eat fresh produce throughout the whole chain. The industrial implementation of research findings was clearly stated and planned including process optimization and management procedures. The expected interactions among partners inside and outside the consortium were widely respected, thus giving a multidisciplinary approach (Fig. 21). The activity was related to the results obtained within the first four WPs of the project, by interfacing diagnostic kit (WP1), process control (WP2), decision support (WP3), innovative processes and management system implementations (WP4). Therefore, WP5 was articulated in four tasks, from Task 5.1 to Task 5.4. In particular, in Task 5.1 involving UNIFG, AUA, and UMIL with AGRONOMIA and Nuvifruits, as partners and the Institute Zooprofilattico per la Puglia e Basilicata (ISZPB) as public Health Institution. The demonstration activity was aimed to give to the SMEs the opportunity to innovate their quality management. The minimal instrument and tools were tested for making as easier as possible the use of the protein antibody arrays for quality evaluation and microbial contamination. The demonstration was focused on the bacteria detection (Listeria monocytogenes and E. coli) using an ELISA based method developed in the WP1. This method compared with the standard procedures allowed gaining detection time less than 24 h, while the standard methods take 4-5 days. The implementation of the developed diagnostic kits allowed checking the quality of dubious fresh-cut vegetables, before the transporation of the final product, in order to guarantee the consumers safety and quality of the products. In addition the application of a quantitative Real-Time-PCR (Q-rtPCR) method associated to MPN for pathogen detection and enumeration in fresh-cut products was also demonstrated with the cooperation of ISZPB. Task 5.2 was aimed to assess the efficacy of process control by the involvement of UNIFG, CU and UCP with AGRONOMIA, Nuvifruits and am additional company external to the project. Quality attributes of rocket leaves and melons were monitored during distribution by temperature data loggers, so to have a thermal history of the products. Starting from the external attributes, internal attributes were predicted by applying the models, and then verified through the application of standard destructive methodologies. Analysis methods for remote collection of volatiles were also demonstrated. The evaluation of volatile profiles associated with deterioration under different storage conditions in relation to appearance, progression of degradation in terms of growth of spoilage organisms, nutritional and organoleptic quality was demonstrated with the objective to complement and expand the destructive diagnostic methods. The VOC demonstration used TD-GC-TOF-MS for assessing quality of fresh-cut rocket salad and melon at the start of processing and during shelf life of the processed products. Task 5.3 related to demonstration of decision support involved Unifg, UCP, ARO, LMET and DARe with Nuvifruits, AGRONOMIA and EFI as industrial partners. Models to assess the benefits of MAP on fresh-cut fruit and vegetables and models to predict the barrier properties of packaging developed in WP3 were validated by using melons from Nuvifruits and rocket salad from AGRONOMIA. In both cases, different steps were carried out and in each of them the main quality parameters were assessed. As regard to model-driven control of spoilage and off-flavor in RTE fresh-cut fruit products, the efficacy of micro-perforated active modified atmosphere (MAMA) packaging for preservation of fresh-cut melons was demonstrated with success at the commercial fresh-cut facility of EFI. The activity of LMET was aimed to determine single cell lag phase durations of pathogenic organisms, e.g. Escherichia coli and Listeria monocytogenes, and to apply statistical methods to distributions of single cell lag times in order to develop models that more accurately predict lag phase in fresh-cut products. The developed models were demonstrated to be suitable tools for decision-making in the food industry. The application of a multi attribute model for investment decisional support through an user-friendly software, developed in Task 5.3 was carried out by direct demonstration through DARe personnel and indirect demonstration through an online manual. In this activity industrial partners within QUAFETY and other SMEs were involved. The main result was the application of the software in investment decision routine, use of the software for analyzing farms issue beyond financial aspects for investments and inclusion of QUAFETY know-how into farms. The activity of Task 5.4 was aimed to demonstrate the innovative processes developed by UNIFG, AUA, INSAD and ARO to the industrial sector by involving EFI and AGRONOMIA as project partners and Farm De Ceglia as external company. The efficacy of studied solutions to increase quality and safety of fresh-cut products was demonstrated. Particularly, benefits of soilless cultivation protocol applied on leafy vegetables were underlined, (raw material with high quality and safety, with higher shelf-life, with a better esthetic value and with a better nutritional value), thus validating under real farm-scale conditions the results obtained in the experimental-scale. The effectiveness of developed waste-water sanitization system as chlorine-alternative method was assessed in the company AGRONOMIA. The efficacy of UV treatment of fresh-cut melons microbial load was demonstrated at commercial fresh-cut facility of EFI. The partners involved were ARO and EFI. The effectiveness of biofilm inhibitors and biofilm removing agents on Listeria monocytogenes was demonstrated by UNIFG. Demonstration was carried out by using biocide on L. monocytogenes able to form biofilm on industrial surfaces, to give a protocol related to “the best practices” to be followed in order to reduce biofilm formation during fresh-cut production and risk of biofilm harbouring L. monocytogenes. The partner INSAD was involved in demonstration of hot water treatment applied to rocket to prolong the shelf life. In the same task the suitability of developed active and intelligent packaging applied to rocket salad was assessed based on quality and safety criteria.
S&T results on Evaluation of economic viability and sustainability of the applied methodologies (WP6)
WP6 focused on testing the hypotheses of ready to eat fruits and vegetables as substitutes for conventional whole products, and quality attributes deriving from innovative production technologies, within those produced by QUAFETY research, in order to understand consumer preferences and draw consideration on the consequent consumer acceptability and measuring consumer perception towards these attributes. In addition, consumers’ acceptability measures were used for setting up the cost-benefit analysis of the innovations produced by QUAFETY. In this WP all partners were involved.
This WP was divided into three tasks. Task 6.1 (first year activities) includes all the activities leading to the estimation fresh cut mathematical functions of consumer preferences, allowing the prediction of consumption behavior towards RTE fruit and vegetables at different levels of price/quality/convenience. This result allows inferring on consumer preferences, consumer perception and provide information to orient applied research and investors choices. Task 6.2 (second year activities) consisted into estimating willingness to pay for different quality and convenience levels. Finally, Task 6.3 (third year of activities) focused on the assessment of the impact on costs and revenues of the new technologies coming out from QUAFETY project.
In order to depict the quality attributes enhanced by the innovations produced within the project, from a consumption perspective, in depth interviews have been carried out within the Task 6.1. QUAFETY Partners have been involved in a survey in which specific and in-depth questions have been asked in order to contextualize the outcomes of their project research in a future real product (concept), in which those innovations are embedded into it, and perceived by consumers. Once answers from research partners have been collected, data have been analyzed and discussed in a focus group environment among researchers of D.A.Re. The discussion, aimed to translate research products into quality attributes, labels, price and shelf life variations, generated the list of attributes and levels to be analyzed in order to build a mathematical function that predicts consumer choices towards RTE products.
Having the attributes, an experimental design was built in order to propose a limited number of alternative products to consumers during the subsequent survey. The alternatives represented “real products” together with “conceptual products” from which consumers were asked to choose.
Finally the survey was conducted with a total of 1,462 interviews across some EU Countries. Cooperation among partners has been fundamental for the implementation of the survey and translation/interpretation in local language, given that fresh cut market has its own specificities in each country. More specifically, about 500 consumers were interviewed in Italy, about 250 in Spain, about 250 in Greece and about 500 in UK. Consumers were selected randomly at the exit of the shopping mall across the entire area of the countries (Italy, Greece) or while shopping online (Spain, UK). However, the sample was stratified by age and gender in order to increase the level of representativeness.
Once data were collected an electronic database was input with all the information. Sample descriptive statistics was prepared and the preference predictive model was set up in order to understand consumer perception of different attributes, and to forecast purchase probability. In order to select the best model, several alternative models were tested competitively.
Moreover, cross-country comparison was conducted in order to depict the main differences in habit, preferences and attitudes toward consumption.
In the Task 6.2 the predictive preference model has been tested and improved in order to calculate the best price at which consumers show the strongest preferences. The hypotheses in the WP test whether quality attributes deriving from newer production technologies have impact on consumer acceptability, hence their willingness to pay towards each novel product attribute (Fig. 23). Knowing the impact on consumer willingness to pay of each innovation helped hypothesizing prioritization of innovation based on profitability of the product differentiation obtained by QUAFETY research. As a consequence, the willingness to pay was modelled and calculated, based on consumer survey data and preferences analysis. With WTP measures, based on empirical models of consumer choice prediction, and set up on survey data, consumer response to the quality attributes deriving from new technologies could be measured in order to evaluate the impact on the consumer price responses towards the innovations analyzed in the project. The analysis of the willingness to pay values was also focused on acceptability measures, in order to be sure that selected attributes did not represent an attended attribute, hence did not orient preferences and choices. Finally, cluster analysis has been conducted in order to better identify eventual marketing directions. We found that shelf-life extension did not generate positive consumers’ reactions as well positive willingness to pay, as opposed to safety enhancement and additional information to consumers.
Activity of the Task 6.3 focused on the cost-benefit analysis, which took into consideration results of willingness to pay calculation, together with the results of a questionnaire to researchers, which involved the collaboration of all project members, in order to calculate cost information relative to QUAFETY innovations and measures the level of profitability for each innovation produced within the project. Cooperation has been fundamental for the individualization of the impact on costs of each innovation produced. Therefore, the activities aimed to estimate the impact of innovations on final company profit and furnish economic viability measures to assess the mere business validity of the innovations produced within QUAFETY research. Results have been considered starting from the perspective of two extreme types of representative companies, from cost structure point of view, and consider different price perspective in the medium long run. For instance, there are innovations affecting positively the reduction of spoilage costs, but an increase in depreciation, because they need more fixed investments. Other innovations increase costs relative to personnel training but increase revenues, etc. Meanwhile, when analysing the implementation of innovations, fixed investments are also taken into consideration. Fixed investments, in fact, require an initial very high cost, with a consequent yearly depreciation. The cost-benefit analysis considers also how eventual increase in profits pays this investment back. As a result, many of the investments proposed by the QUAFETY innovations did not get high returns of substantial increases in the net present value of the company. As a consequence, fixed investments are not always desirable. In those cases, researchers should consider the provision of third party service, which will not need the implementation of fixed investments in the company, but will only increase variable costs of a given amount.
Finally, DARe results, obtained with the full collaboration of all partners, allowed to state that not all innovations impacted positively on the willingness to pay and not all innovations generated a gain in total profits. There are innovations that do not impact price positively nor reduce costs, making production more efficient. In those cases, the probability that companies will adopt the innovation is noticeably reduced, while other innovations could effectively contribute to a profit increase, and be adopted for convenience reasons. Thus, the indubitably positive impact of knowledge and quality of research does not always get positive response in terms of company convenience. However, obtained results provided a critical approach to look at industrial scale-up and implementation of research results for profit oriented reasons. Nonetheless, the revision of some of the innovations, with a managerial perspective, could lead to higher level of convenience.
S&T results on Safety & Quality Management System (WP7)
WP 7 was organized into three tasks to develop assessment tools for quality and safety of the SME of the fresh cut sector, to apply and analyse the collected data and finally to propose potential interventions for improvement in the participating SMEs. To achieve these objectives all partners actively participated to the above tasks. The main results of the WP7 were the development of a diagnostic instrument for the assessment of the management systems applied in fresh cut sector related to food quality and safety which include a questionnaire and a safety assessment plan (SAP). The questionnaire consists of five sections. The first includes background information for the SMEs, the second and third information on the risk factors, the fourth related to the QMS/FSMS applied in SMEs, if any, and the last one includes questions for good agricultural practices during pre harvest, prerequisite programmes and processing control points. The responses from the application of the questionnaire followed the process described in Fig. 24. Thus, applying principal component analysis significant components were extracted to provide a score for QMS/FSMS.
The developed SAP was aimed to assess the level of safety along the production-processing chain and the safety level of the products. In this assessment plan, the sampling points were identified, the required number of samples were determined, and the appropriate analytical methods for the sampling and determination of safety and quality level at the different sampling points were indicated. After the application of SAP, the data were collected and analyzed. This SAP will be a key part of the food safety and quality management system that will be used in combination with the questionnaire for the assessment of the overall performance of the QMS/FSMS of the participating SMEs and to draft the recommendations for the improvement of the management systems applied in the SMEs. These will be an integral part of the management systems applied (Fig. 25).
The assessment of the QMS/FSMS revealed a disagreement between the scores from the questionnaire and the scores from the interpretation of the SAP in 2 out of 4 SMEs (Fig. 26). For two SMes very good agreement between the two parts of the diagnostic instrument (questionnaire and SAP) was found: SME C ranked at the “top” level for both parts of the instrument, whereas SME A ranked at the “good” level for both parts of the instrument. On the contrary, disagreement between the two parts of the instrument was observed for the other two SMEs: SME D and SME B were ranked at the “top” level according to the questionnaire and at the “moderate” level according to the interpretation of the SAP. In cases of disagreement between the questionnaire and the SAP, the latter provides more objective evidence of the safety and microbiological quality status of the SME. Very often a bias is inserted in an attempt to ameliorate the real safety status of the SME. In such cases, a detailed auditing will give the opportunity to depict the actual safety status; at the same time training of the personnel is necessary in order to understand the importance of accurately answering in such audits.
The impact on other WPs is defined by incorporation of these tools in the FSMS guide of WP8 and to decide which of the procedures developed in WP1 to WP5 will be useful alternatives to improve the processes used in the fresh cut produce sector. The creation of a FSMS guide is the result of info and data collected by all partners trough the experimental work and demonstration activity in the SMEs. This guide should be a tool for daily use; these guidelines should help fresh-cut industry operators to manage and improve safety and quality of the fresh-cut products. The guide is available in USB-key format for fresh-cut chain operators.
Dissemination (WP8)
The Objective of WP8 was to disseminate project results to potential users (i.e. Food industries, consumers, health institutes, mass media, etc.) by online websites, seminars, training courses, conference attendance, papers, etc. The dissemination activity is the result of a great and continuous cooperation among all Quafety partners.
The objective of Tasks 8.1 and 8.2 consisted in creation of a subsection dedicated to fresh-cut management on all 3 Freshplaza’s websites (English, Spanish, Italian); in this subsection, called Quafety, divulgative articles on fresh-cut produce sector are published to disseminate scientific studies and to create a connection among the play actors of the whole fresh-cut chain. Furthermore, the official website of the project (www.quafety.eu) has been created by Freshplaza, while it is managed by all partners under leader supervision.
For dissemination work, more than 500 articles (Task 8.3) on fresh-cut produce chain were published and are online available at:
1. http://www.freshplaza.com/sector/109/quafety
2. http://www.freshplaza.es/sector/101/Quafety
3. http://www.freshplaza.it/sector/101/Quafety
The dissemination of scientific results, technological news, educational events and training courses of QUAFETY and fresh-cut produce chain will continue also behind the QUAFETY conclusion.
A guide on Food Quality Management System (FQMS) for fresh-cut segment was created and is available in USB-key format (Task 8.4). The guide is a list of 1) good agricultural practices (GAPs), 2) good hygiene practices (GHPs), 3) good post-harvest management practices (GMPs), 4) good processing practices for fresh-cut industry, which are crucial for HACCP (Hazard Analysis and Critical Control Points) system. The guide is the result of info and data collected by all Quafety partners through the experimental work and demonstration activity in the cooperating fresh produce companies.
Freshplaza created ‘QUAFETY group’ on Tacler (www.tacler.com) which is a Fresh Produce (fresh fruit and vegetables) industry web-based community platform. This platform was used as a permanent online network for connecting stakeholders with Quafety scientists (Task 8.5). The permanent interaction between scientific community and industry is in fact crucial for increasing company competitiveness in the global market. Tacler users from all over the world, daily demonstrate their need to enter new markets, to improve their products in terms of quality, safety and added value, to be competitive of economical and environmental sustainability. In Tacler, Freshplaza organized meetings with QUAFETY scientists to create connection with fresh produce chain actors registered to Tacler. This initiative represented the starting point of this new Tacler joint mission, with the idea to continue in the future this type of meetings.
Through dissemination activity, Freshplaza divulgated all info on training and educational courses, conferences and event for moving students, scientists, and fresh-cut chain operators (Task 8.6).
At the project conclusion, Freshplaza organized the final workshop in Rotterdam on 25th March 2015 (Task 8.7). During the workshop all QUAFETY results were showed and a CD-Rom with the most important project documents was created to distribute to the participants.
Potential Impact:
Project QUAFETY has proposed a complex program of food quality & safety solutions throughout the farm-to-fork supply chain for RTE fresh produce. Adoption of these solutions by SME producers may potentially improve their products and thus strengthen their competitiveness in the market. Furthermore, the innovations introduced by QUAFETY may improve sustainability of the technological process by substituting the use of hazardous and environment-polluting chemical sanitizers.
As the category of RTE products is clearly consumer-oriented, all solutions proposed by QUAFETY also address the needs of consumers that due to time limits or other circumstances choose not to prepare meals “from scratch”. RTE products are intended for direct consumption and so the acceptance of these products by consumers depends on their trust in measures undertaken by producers to ensure the product safety and quality. An increased level of confidence will go in line with the task of promoting fruit and vegetable consumption in Europe in order to reduce the risk of obesity and chronic diseases (European Parliament resolution 2010/C 8 E/18).
In addition QUAFETY results provided scientific evidences which may be exploited by health authorities (at EU, national, and regional level) in order to evaluate whether further regulation is required in this area of food safety and to support educational campaigns for healthy nutrition.
Another most important impact of the Project is associated to its function as a reference point for specialized and general media when covering quality and safety aspects of RTE fresh produce. As in the last few years fresh-cut products have been the target of a number of media attacks the presence of QUAFETY evidences has contributed to a more equilibrated approach to the topic of food safety.
Finally, last but not least, as a large of information coming out from a R&D project like QUAFETY are basic scientific information (representing major or minor breakthrough in the knowledge on specific topics), it has largely impacted with the scientific community throughout a number of scientific communications either presented in Conferences or published on peer-reviewed Journals.
Main “impacting” aspects of the Project are listed below by workpackage, while a summary of main dissemination numbers will be described thereafter.
Potential impact for WP1
The results obtained on the MPN-QPCR methods for quantification of L. monocytogenes and E. coli O157:H7 from fresh cut vegetables and enrichment free methods can enhance the safety of the whole fresh cut market.
Pathogen prevalence in fresh cut fruits and vegetables is essential for risk assessment protocols. This information along with the measure of genotypic diversity should be taken into consideration when designing intervention strategies aiming at the safety improvement of the fresh cut sector. At the same time information on gene transcription may improve our understanding of the pathogens physiology that may contribute to the development of more efficient tools for their control.
The antibody array containing specific antibodies against the senescence associated proteins and against the human pathogen bacteria allow the evaluation of quality and safety of the produce in fast and reliable way, at any step of the whole distribution chain from the grower to the consumer. The idea is that with a simple sample homogenization the crude protein extract can be used to hybridize the antibody arrays and then with a scanner highlight the presence of protein associated with quality losses or the presence of harmful bacteria (deliverable D1.3 UMIL). This diagnostic kit can enhance the quality and the safety of the whole fresh cut market.
Potential impact for WP2
The results of Task 2.1 provided useful applications at different levels of the research. The photographic scale developed in Deliverable 2.6 provides an objective tool for the evaluation of appearance of rocket and fresh-cut melons, which can be used for the evaluation of raw material both in research and by companies. The degradation kinetics in packaged samples both in isothermal and non-isothermal conditions were obtained, as reported in Deliverable 2.7. The kinetics may help processors to understand the most critical factors for shelf-life and specifically the impact of temperature. As complementary results the objective of an accurate shelf-life prediction, taking also into account the fate of nutritional components, was achieved. Finally the models relating changes of internal nutrients to appearance degradation may be a useful tool for having information on nutritional content, without the use of sophisticated methods and instruments.
Results obtained in Tasks 2.2. and 2.3 allow the definition of product shelf-life concerning their bioactive properties and protective health effects. Also, the studies undertaken in these tasks allowed the development of an expert-system available for the industry to predict the depreciation of the functional properties over time under different processing conditions. (http://www.quafety.eu/index.asp?partner=1&news=171). The “Expert System” program gathers, transfers and gives worldwide access to the knowledge obtained throughout QUAFETY Project on nutritional quality of processed rocket leaves, melons and strawberries as affected by processing (production process; cutting degree) and storage conditions (package type, modified atmosphere and storage temperature). It thus provides the industry with useful information for developing processes and making management decisions on the modulation of nutritional quality and shelf-life extension.
Task 2.4 generated useful baseline data on VOC profiles for melon and rocket (Deliverable 2.11) as well as for strawberries. It also identified potential markers based on VOC analysis of use to the processing industry for assessing processes which affect shelf-life such as mechanical damage and changes in temperature control as well as the quality of incoming material for processing (Deliverable 2.12). Interactions between CU/Markes and UK processing companies indicate a strong interest in taking this forward. The interaction with UCP also linked the VOC markers to sensorial data and analyses of compounds of nutritional significance thus VOCs could also be used to assess changes in processing which may also be affecting nutritional value and or shelf-life. The potential of VOC analyses for detection of contamination with human pathogens would also be of benefit to the industry as a complementary approach to other methods that are able to detect very low levels of contamination more rapidly that the culturing methods currently used.
Potential impact for WP3
The results of Task 3.1 provided a decision-support information for operators (growers and processors) choosing genetic material for producing high-quality RTE melon products. It will guide breeders in creating new melon genotypes for fresh-cut processing. For consumers, knowing a genetic origin of RTE melon may indicate a chance of off-flavour development during storage.
The kinetics determined in the Task 3.2. allowed the prediction of quality changes of rocket leaves and fresh-cut melon and strawberries during storage. The integration of all these results contributes to the understanding of the effects of storage conditions on overall quality of RTE vegetables and fruits and provide useful information for developing processes. Aiming at the prediction and modulation of nutritional quality and shelf-life, these kinetic parameters and physiology-based models are adequate for the development of tools as predictive softwares for the industry.
As for Task 3.3. models whose parameters can be simply obtained and that can be easily applied to different polymeric film are useful to optimize the packaging and the production process according to the packaged product features. Moreover, the respiration rate modeling is central to design modified atmosphere package intended for fresh-cut fruit and vegetables. Models of WP3 were used to optimize the packaging conditions for fresh-cut melon from Nuvifruits.
The results of Task 3.4. will provide a decision-support information for processors choosing optimal packaging solutions for RTE fruit products. For consumers, the optimized packages will provide products with extended storage life and improved sensory quality.
In Task 3.5 the survival of L. monocytogenes and E. coli after subjection to mild heat treatment and acidic conditions was studied (D3.18). The outcome of this may be of value to processors of fresh-cut foods, since it will provide information to improve processing procedures, thereby increasing safety of the product. In addition, the individual lag times of sublethally heated and acid stressed L. monocytogenes and E. coli at a range of recovery temperatures were estimated and a model was developed. This will provide a valuable tool for the food industry enable decision making and risk assessment, thus contributing to manufacture of safe products through understanding ways of controlling L. monocytogenes and E. coli (D3.19).
The software developed by DARe in Task 3.6. being easy to send to companies, is a effective way to communicate QUAFETY results to companies and increase the probability of further scaling up activities.
Potential impact for WP4
The results of Task 4.1 will be applied by growers and allow them increasing yield and improving quality of leafy vegetables with rational use of water and fertilizers. Furthermore, the new method to control the nitrate level in the vegetables will allow the growers to promote their produce as more consumer-friendly and healthy. It is important that the results have been already tested at a bigger scale at a commercial farm and the conclusions have been confirmed. For consumers, it will provide cleaner and safer foodstuffs with lower nitrate content, better nutritional quality (higher vitamin C) and slower spoilage. The latter improvement will be important also for the retailers. Thus, the development will contribute to the reduction of food losses. For policy makers, it will provide a guideline for permissible level of nitrates in soilless-grown leafy vegetables. The findings of this applied work may provide basis for a more basic research dealing with regulation of plant development and metabolism by mineral nutrition, and for breeding crops better suited to soilless cultivation.
The results of Task 4.2 are primarily important for ensuring microbiological and chemical safety of washed and ready to eat leafy greens. Prevention of foodborne disease outbreaks is equally important for the consumers and for the producers, as well as for the public health authorities. These results suggest the feasibility of substituting hypochlorite with the more effective chlorine dioxide which at low doses can reduce the risk of product contamination with pathogenic bacteria (e.g. Salmonella, E. coli H157:O7, Listeria) and with spoilage bacteria. In addition, it will prevent generation of potentially harmful and carcinogenic decontamination byproducts such as trihalomethanes in the products and in the plant wastes. The ultrafiltration technique permits to reuse the water in the process and to improve process sustainability. As an additional effect the system allows to recover some of the water used for washing, and to put it back on circulation reducing the environmental impact of the whole system being an important result for the producers and for the public and for the authorities. It is especially important that the developer of this technique is a commercial processer so that the technique has been tested at industrial-scale facility.
The results of Task 4.3 are primarily important for ensuring microbiological and chemical safety of fresh-cut melons. Prevention of foodborne disease outbreaks is equally important for the consumers and for the producers, as well as for the public health authorities. Substituting hypochlorite wash with an efficient physical thermal treatment leaving no toxic residues will reduce the risk of product contamination with pathogenic bacteria (e.g. Salmonella, E. coli H157:O7, Listeria) and with spoilage microorganisms (bacteria, yeasts and moulds). On the other hand, the treatment will make the preparation of melons for fresh-cut processing more sustainable and environmentally friendly by reducing water and energy consumption and preventing the release of toxic and carcinogenic byproducts such as trihalomehanes. The reduction of environment pollution with decontamination byproducts is important for fruit processors, authorities and public.
The results of Task 4.4 are primarily important for ensuring microbiological safety of food industry in general and fresh-cut industry in particular. Listeria is a highly dangerous potentially lethal pathogen so that prevention of its outbreaks is equally important for the consumers, for the producers and for the public health authorities. Biofilms of Listeria result in cross-contamination of the produce with this deadly pathogen. They are very hard to eradicate if not treated on time and can result in closure of a factory by health authorities. The work resulted in preparation and dissemination of recommendations to the industry for practical measures on preventing the buildup of Listeria biofilms on the surfaces of fresh-cut facility. These recommendations are important also for policy makers for ensuring public health.
The results of Task 4.5 will be of primary importance for fruit processors due to the increasing efficacy of cutting and peeling operations. It will improve work conditions and the safety of the technological process by preventing manual operations with sharp knives under worker-unfriendly conditions (low temperature, decontamination chemicals). Exclusion of a human factor as a potential source of contamination will improve the product microbiological safety. On the other hand, substitution of manual operations with automatic ones will allow running them under strict conditions e.g. under UV light and low temperature further reducing the chances of product microbial contamination with spoilage or pathogenic microorganisms. The latter factor is important for processors, consumers and health authorities. The work has resulted in submitting a provisional patent application that has already attracted the interest of machine-building industry that is another important stakeholder in the fresh-cut technology.
The results of Task 4.6 will improve quality and reduce spoilage of ready-to-eat products, in particular shredded Chinese cabbage. These results are important for vegetable processors, retailers and consumers. Controlling the product spoilage will contribute to the global task of reducing food losses. For researchers the findings of this applied work may provide basis for a more basic research dealing with regulation of senescence and metabolism in plant tissues by physical factors such as temperature.
The results of Task 4.7 are primarily important for ensuring microbiological and chemical safety of ready-to-eat leafy greens and fresh-cut fruits e.g. melons. Prevention of foodborne disease outbreaks is equally important for the consumers and for the producers, as well as for the public health authorities. Strengthening antimicrobial measures by additional hurdle of physical intervention (UV light) immediately before packaging will reduce the risk of product contamination with pathogenic bacteria (e.g. Salmonella, E. coli H157:O7, Listeria) and with spoilage microorganisms (bacteria, yeasts and moulds). The latter factor may be important for both retailers and consumers and contribute to the global task of reducing food losses. For researchers, the findings of this applied work may provide basis for a more basic research dealing with regulation of metabolism in plant tissues and modulation of host-pathogen interactions by physical factors such as UV light.
The results of Task 4.8 are important for ensuring microbiological safety and preservation of quality of ready-to-eat fruit and vegetable products (leafy greens and fruit) and therefore are of special importance for the processors. Prevention of foodborne disease outbreaks is important for the consumers and for the producers, as well as for the public health authorities. The development of active antimicrobial packaging and coating will reduce the risk of product contamination with pathogenic and with spoilage microorganisms (bacteria, yeasts and moulds). The latter factor may be important for both retailers and consumers and contribute to the global task of reducing food losses. The novel Time-Temperature indicator may be of interest for retailers, consumers and decision makers providing a more accurate instrument to evaluate the goodness of the product on the shelves. It will prevent both unnecessary rejections and wastage, on one hand, and keeping the unmarketable products on the shelves, on the other.
The results of Task 4.9 will allow the maintenance of cool conditions throughout the supply chain, from the field and to the final consumer and therefore will be of interest for all stakeholders: growers, processors, distributors, retailers and the consumers. The containers developed in this task will improve quality preservation and reduce proliferation of microorganisms on leafy greens eventually controlling their spoilage. This will contribute to the global task of reducing food losses. Controlling microbial proliferation will prevent the potential pathogens of reaching an infective dose and this will improve the produce safety reducing the risk of disease outbreaks. It is especially important that the developer of the container is a commercial processer so that the appliance has been tested under industrial conditions.
Potential impact for WP5
WP5 represents the most relevant WP in regard to the impact of main results of QUAFETY project. In fact in this WP a number among the most promising results from previous WPs were tried at a larger scale and often their application was demonstrated at industrial sites, including processors internal to the Consortium, and a number of external entities, including private producers and processors, and public health authorities. Apart from being a necessary step toward potential implementation of the innovations into the “real world” the other important thing is that participants in this Project were the first to whom results were disseminated, and in the most effective way. In this way results dissemination/exploitation started within the Consortium and this in turn have enriched the same results with feedbacks deriving from this first phase of “internal” dissemination. In addition it is worth noting that in sectors that are disjointed as the agricultural and the food system, the introduction of a systemic approach for food quality and safety is of strict relevance. Furthermore, WP5 highlights the importance of relations between the different components of the system. The cohesion between the different organizational objectives is achieved only by acting with a system able to ensure the integration of the activities carried out by different actors.
Potential impact and main dissemination of WP6
WP6 focused on testing the hypotheses of ready to eat fruits and vegetables as substitutes for conventional whole products, and quality attributes deriving from innovative production technologies, within those produced by QUAFETY research, in order to understand consumer preferences and draw consideration on the consequent consumer acceptability and measuring consumer perception towards these attributes. In addition, consumer acceptability measures were used for setting up the cost-benefit analysis of the innovations produced by QUAFETY. Thanks to these activities DARe staff were in touch with a series of companies which showed deep interest into the economic evaluation of the innovations. They most common statement has been “usually we implement innovations and make things work out, with such approach we have a prevision of what would be the impact on our cost structure and price potentiality”. The main impact of this activity is related to the possibility to use this approach to enhance communication between scientists and stakeholder, and to further develop this tool to drive innovation processes.
Potential impact and main dissemination WP7
The WP7 results are addressed to four basic groups of users:
1. Stakeholders from the food sector as end-users of final application including: food industry, consultancy services, retail business
2. Public food safety authorities
3. Researchers in the field of food science, agribusiness and horticulture
4. ICT companies for the development of new applications for agriculture and food sector, mainly using implemented tools, encompassing: SME developers of ICT for agriculture platform and technology producers.
The food sector is of strategic importance for European society and economy. Due to its complexity, food operators have to manage many different and heterogeneous sources of information. The tool will help fresh cut food sector to collect, storage, share and analyse data related to quality and food safety. The potential impact of application the specific tools developed in WP7 that is, the questionnaire and the safety assessment plan (SAP), as an integral part of the quality management and food safety management system in a SME is of great importance. These could be used as a self-assessment tool in the SMEs and as an internal audit tool, providing specific quality tools to the fresh cut produce sector for their reviewing managerial processes. Results obtained by the SAP will determine the actual hygiene and safety level of the SMEs and will validate the best practice score that will be assigned to SME after interviewing. The socio-economic impact is that SMEs will improve their quality and food safety level via intervention in conjunction with the procedures developed in the project and incorporated in the FSMS guide. The societal implication is evident by offering fresh produce of a better quality and safety to the final consumer.
Main dissemination numbers
Dissemination was one of the strong points of Project. QUAFETY activities and results have been presented at various level in a very hign number of Countries, both in Europe (Fig. 28) and outside (Fig. 29). With reference to the previously described results in WP8, following a synthesis of main dissemination numbers are reported according to the type of stakeholder target (R&D, industry and authorities, media and consumers).
Dissemination activity targeted for R&D. Up to date a total of 24 peer-reviewed scientific papers have been published within journals, edited books, and conference proceedings; of these, 15 proceeded from activities in WP1, 4 from activities in WP2, 1 from those of WP3, and 4 from those of WP4. A prudential estimation considers at least 20 more papers being published within the next 12-18 months mainly from WPs 2, 3, and 4. In addition a total of 13 dissertation Thesis have been produced within the Project, mainly at AUA and UNIFG.
Participation of QUAFETY scientist in international scientific conferences allowed the presentation of 48 oral contributions (11 of which as invited keynote) and a total of 44 posters.
Finally, “special QUAFETY sessions” have been organized within 4 major scientific conferences in Italy, Portugal, and Poland.
Dissemination activity targeted for industry and authorities. A very large effort was made in order to diffuse the outcomes from the Project to the produce industry, with particular emphasis to fresh-cut produce companies. With this objective a number of strategies were used. More than 500 articles published in the technical and popular press and websites, mainly in English, Spanish, and Italian. In addition results were illustrated in about 55 oral presentations in industry meetings.
QUAFETY scientists participated as instructors in industry targeted educational events in Turkey and in UK while 4 specific workshops were organized for a public audience in Italy, Greece and the Netherlands, and within the popular international exhibition Fruit Logistica in Germany.
In some of these events informative material was freely distributed to participants in form of flyers, USB pendrives, and CD-ROM.
Additional dissemination activities were performed through the world wide web with the project website (Fig. 30) and the fresh-produce industry interactive platform Tacler (www.tacler.com).
Dissemination activity targeted for media and general public. Besides the QUAFETY website, a Youtube channel was available to download video documents produced within the Project. In addition UNIFG Facebook page hosted all kind of news related to the Project for about 1,300 contacts.
Finally a number of interviews were released by QUAFETY scientists to different media type, both web-based and traditional, at local, national, and international level.
List of Websites:
Project website
www.quafety.eu
Project Coordinator (Prof. Giancarlo Colelli)
giancarlo.colelli@unifg.it
The Project QUAFETY (Comprehensive Approach to Enhance Quality and Safety of Ready to eat Fresh Products) was aimed to improve quality and safety of Ready-to-eat (RTE) fresh produce throughout the whole chain by developing new predictive and probabilistic models and decision-making tools, by exploring rapid and non-destructive methods for quality evaluation and prediction, and by experimenting novel technologies, in order to quantify and manage spoilage and pathogen microorganisms, minimize risks to consumers, and preserve quality. The participants were 14, from 7 Countries and included 6 SMEs, 2 public research Institutes and 6 Universities, assorted in terms of scientific and technological expertise. The presence of SMEs ensured the exploitation of the results directly and indirectly.
As a response to the industry needs and in relation to the state of the art, within the project QUAFETY the following results were obtained:
- reliable and fast detection diagnostic kit based on key molecular probes for L. monocytogenes and E. coli O157:H7 and O104:H4, and on specific antibodies against the senescence associated proteins and against the human pathogen bacteria to detect stress conditions and presence of contaminants;
- rapid non destructive tools for process control which were aimed to better estimate shelf-life of fresh-cut products and to link the deterioration of external quality indexes to nutritional attributes, to identify nutritional, functional, and quality markers including volatiles to be monitored during process to reduce the impact of processing on final quality;
- decisions supporting tools to allow industry operators to understand and implement process innovations aimed to improve safety and quality, and particularly to select an “ideal melon” for processing, to predict barrier properties of polymeric films and the oxygen evolution in MA packaging, to estimate the respiration rate of fresh-cut packaged products and the impact of temperature and cutting on respiration kinetics, to predict bacterial lag time at various temperatures and finally to evaluate innovations starting from QUAFETY results which were included in an excel spreadsheet based software to support innovation investments;
- innovative processes and technologies were developed to improve quality and safety of the final products starting from raw material quality from pre-harvest (management of nutrient, varietal selection) to postharvest (passive refrigeration system, developing an automatic hands-off machine for melons) and processing (decontamination systems for water, products and industry plants, use of physical treatments and innovative packaging solutions);
- evaluation of the impact on markets and profitability for the industry considering consumer response to safety and quality attributes deriving from the potential application of QUAFETY innovation;
- development of a more effective and efficient Quality/Safety Management System for RTE fresh produce industry.;
The results of this project contribute to provide scientific evidences on factors affecting quality and safety of fresh-cut products with the aim of ensuring high quality standard throughout the chain. These results may be a reference point to mass media, and eventually to EC and other health authorities (also for campaigns for healthy nutrition), which will contribute to increase consumer awareness on quality and safety of fresh-cut products. Finally the level of innovation reached with the project will strengthen industry competitiveness.
Project Context and Objectives:
An important category of RTE food is represented by fresh-cut fruit and vegetables (F&V). The fresh-cut sector may be the smallest and most recent F&V segment in Europe, but it is clearly the fastest growing market. Fresh-cut salads are the favourite RTE fresh produce of European consumers, with more than 50% of the market volume, while fresh-cut fruits account for more than 10% of the share. These products meet new consumer's demands for healthy and easy to prepare food. However several outbreaks of food-borne diseases linked to the consumption of RTE produce have been recorded in past years.
One important thing related to safety of RTE fresh produce is that the preparation process does not include a “killing step”, or a treatment which reduce the final microbial count to value close to zero. This indicates that fresh-cut vegetables sold in the market may represent an underestimated public health risk due to the possible presence of pathogenic bacteria. Another peculiarity of RTE fresh F&V is that the food is constituted by living cell tissue, rapidly metabolizing, especially when peeled and cut in portions for higher convenience. Quality attributes (i.e. appearance, texture, flavour, and nutritional value) degrades very fast and shelf life is often a matter of days. For all these reasons the need of improvements in terms of safety and quality of fresh-cut produce is of paramount importance.
A number of priorities are on top of the list which may have a direct influence on RTE food consumers. SAFETY of course, not only in relation to microbial risks, but also related to other possible chemical contaminations, both from anthropogenic activities or naturally occurring. And QUALITY, especially considering expectations from consumers who pay a relatively high price in order to purchase a high-convenience fresh fruit or vegetable.
As a response to the industry needs and in relation to the state of the art, QUAFETY aims to improve safety and quality of RTE fresh produce throughout the whole chain, addressing the following objectives:
- to provide the industry with diagnostic kits for the evaluation of microbial contamination and shelf-life;
- to provide useful tools for process control to the fresh-cut industry based on non-destructive and rapid measurements, aimed to enhance the final quality of the product;
- to provide the industry operators with decisions supporting tools in very critical points of the fresh-cut processing chain from raw genotype selection to economic strategic planning- to improve, develop and implement process innovations aimed to improve safety and quality;
- to consider consumers’ response to safety and quality attributes deriving from the novel applications in order to evaluate the impact on markets and profitability for the industry;
- to design and implement a more effective and efficient Quality/Safety Management System for RTE fresh produce industry;
- to disseminate in a modern and efficient way all the innovative products of the present proposal.
The objectives of the proposal were reached through the realization of 9 Work Packages (WP) strongly interconnected, as shown in the conceptual diagram (Fig. 1). WP1 developed diagnostic kits, WP2 was dedicated to process control aids, WP3 developed decision support tools, WP4 investigated innovative processes, whereas technological innovations went through implementation and demonstration in WP5 and economic evaluation in WP6. All results represented valuable information for a more efficient management system for quality and safety (WP7) disseminated to potential users in WP8; WP9 dealt with the management of the consortium.
The objective of WP1 was to develop diagnostic kits for the detection of microbial contamination and the quality losses, reached through 4 tasks carried out from 3 different partners. The aim of Tasks 1.1 and 1.2 was to develop fast and reliable detection methods of L. monocytogenes and E. coli O157:H7, also through the individuation of key molecular probes able to detect genes involved in pathogenicity. The third task (Task 1.3) reported the research activities focused on the development of antibodies against the pathogens involved in major outbreaks of foodborne diseases in order to detect the presence of contaminants, while Task 1.4 was related to the individuation of molecular markers linked to physiological degradation of the produce, such as lipid peroxidation, ascorbic acid and chlorophyll degradation. During the second period, according to the recommendations received by the EC reviewers, researchers involved in this WP extended the validation of PCR methods developed during the first period for L. monocytogenes and E. coli O157:H7 and qPCR method for E. coli O104:H4, on non-artificially-inoculated samples, and also evaluated the prevalence of L. monocytogenes and E. coli O157:H7 in melons.
The objective of WP2 was to provide to the fresh-cut industry useful tools for process control based on non-destructive and rapid measurements or evaluation, aimed to enhance the final quality of the product, and it was organized into four tasks. Task 2.1 was aimed to study degradation kinetics of quality attributes for fresh-cut melons and for rocket leaves, identifying relationships between the degradation of external and internal quality attributes and validating the mathematical model developed. Task 2.2 was mainly focused on the determination of relevant nutritional and functional quality markers for fresh-cut melon, strawberries and ready-to-eat rocket leaves and on the optimization, selection and implementation of the most efficient techniques to assess these representative markers (chemical and/or biochemical markers). Moreover it was intended to determine throughout storage the effect of different processing and storage conditions on the quality markers, namely cut sizes, modified atmosphere packaging and storage temperature. The objective of Task 2.3 was to study the effect of different processing steps on quality markers (determined in Task 2.2) on functional and nutritional properties. Task 2.4 was aimed to study the volatile organic compound (VOC) profiles associated with cantaloupe melon and rocket salad, and to assess profiles of samples from other partners, to individuate optimal pre-and postharvest handling conditions. After the midterm report, the EC reviewers requested to extend the VOCs study also to strawberries as non-destructive markers for quality and safety. Therefore, a further objective was to obtain preliminary data for whole and halved strawberries using the TD-GC-MS TOF system.
The general objectives of WP3 were to provide tools for the industry operator to support decisions in very critical points of the fresh-cut processing chain from raw genotype selection to planning economic strategy for investments. The WP3 was articulated in 6 tasks, each one with a different objective. Task 3.1 was aimed to define morphological, physiological and biochemical parameters of an “ideal melon” most suited for fresh-cut processing. The objective of Task 3.2 was to determine the kinetics of respiration of fresh-cut fruit and vegetables as a function of oxygen partial pressure, and to evaluate the effect of temperature and cutting on these kinetics, developing a physiological model to estimate the potential benefits of MAP on melon and rocket leaves. Task 3.3 was related to the development of mathematical models to predict the barrier properties of polymeric films and the respiration rate of fresh-cut packaged products. The objective of Task 3.4 was to build a user-friendly decision-support tool for predicting oxygen dynamics in MA packages and to test its performance with fresh-cut melons. The main objective of Task 3.5 was to determine the single cell lag phase duration of undamaged and sub-lethally damaged (by heat and acid) pathogenic bacteria (Listeria. monocytogenes and Escherichia coli) and develop a model to predict lag time at various temperatures. Finally Task 3.6 wanted to provide a decision support tool enabling industry to better understand innovations produced within QUAFETY and select those coherent with their prioritization in terms of innovation needs and minimum requirements for the scaling-up.
WP4 was aimed to develop innovative processes to improve quality and safety of the final products, it was led by ARO and organized into 9 tasks.
The purpose of Task 4.1 was to obtain in several leafy vegetable species a raw material for fresh-cut processing with improved characteristics in terms of safety, nutritional quality and shelf-life, by strategically managing the nutrient solutions in different soilless cultivation systems, including the definition of innovative optimized growing protocols for several model species as lettuce, endive, escarole and rocket salad.
Task 4.2 was related to the implementation of chlorine-alternative method for water and product disinfection. The goal of Task 4.3 was to improve the efficacy of decontamination of raw melons intended for fresh-cut processing in order to ensure microbiological safety and to control spoilage microorganisms on ready-to-eat products. The objective of Task 4.4 was to analyze the main critical factors affecting the formation of Listeria monocytogenes biofilms in the fresh-cut industry and to find the efficient approaches to its control. The purpose of Task 4.5 was to improve efficacy and safety of fresh-cut melon and watermelon processing by substituting manual peeling and cutting operations by an automatic hands-off machine. Task 4.6 was aimed at testing a novel postharvest heat shock treatment as means for improving quality and safety of fresh-cut fruit and vegetables and extending their shelf life. The objective of Task 4.7 was to examine the efficacy of ultraviolet (UV) light for improving quality and safety of fresh cut products. Task 4.8 was aimed at addressing two types of packaging: (a) active and (b) intelligent. The active packaging was based on active antimicrobial compounds using two methods of their release to the product surface: (i) from the packaging through the headspace and (ii) via edible coating. Regarding the intelligent packaging, the work aimed developing a novel Time-Temperature Indicator of the product freshness. The general objective of Task 4.9 was to ensure the cold chain maintenance by applying a Passive Refrigeration System (PRS), transportable and stackable, to be used for fresh-cut products throughout the supply chain.
The WP5 was mainly aimed to demonstrate the efficacy of the developed technologies to ensure safety and quality of ready-to-eat fresh produce throughout the whole chain. The activities were articulated in four tasks, one for each of the previous WPS, Task 5.1 (Demonstration activities for diagnostic kits), Task 5.2 (demonstration of process control systems), Task 5.3 (Demonstration of decision support tools) and Task 5.4. (Demonstration of innovative processes). Therefore, demonstration was aimed to prove the technical viability of the new technologies, thus expecting to speed up the adoption of the developed innovative tools at the industrial level.
The WP6 was aimed to reach two main objectives. The first referred to the modelling and calculation of willingness to pay, based on consumer survey data and preferences analysis (Task 6.1). The second was related to the cost-benefit analysis, which took into consideration the results of willingness to pay, together with the results of a questionnaire filled by QUAFETY investigators (Tasks 6.2 and 6.3) in order to calculate cost information relative to deriving innovations and measure the level of profitability for each innovation produced within the project.
WP7 objectives were aimed to design tools to support the implementation of an effective and efficient Quality and Food Safety Management Systems in the fresh-cut produce sector. These tools included the development of a validated diagnostic instrument (questionnaire) to evaluate best practices in SMEs and the design of a Safety Assessment Plan (SAP), developed in the Task 7.1 to determine food safety along the production – processing chain. This SAP protocol was used during a pilot test, involving several SMEs within the Task 7.2 and finally the results of the questionnaire addressing technological and managerial elements critical to QMS/FSMSs and of the SAP were critically compared to assess the overall performance of SMEs and its potential improvement in the Task 7.3.
Finally one of the general objective of the project very important for all partners was related to the dissemination of the results obtained (WP8). The 7 tasks of WP8 were aimed to disseminate the project results to potential users (i.e. food industry, consumers, health authorities, mass media) during the project lifetime, through the project website (www.quafety.eu) organization and participation to events, papers and divulgative articles, and in particular through the capillary distribution of the online newsletters and websites of FreshPlaza (www.freshplaza.com one of the most visited websites dealing with postharvest technology and innovation, published in English, Spanish and Italian), currently used as an indispensable source of daily news among the professionals which operate in the fresh produce industry.
Project Results:
S&T results on diagnostic kits (WP1)
The aim of the research activities carried out in the WP1 were oriented on the identification of reliable and fast diagnostic kits able to detect the presence of human pathogen bacteria such as Escherichia coli and Listeria monocytogenes and assess the quality of fresh cut vegetables. In this WP were involved the University of Foggia (UNIFG), the Agricultural University of Athens (AUA) and University of Milano (UMIL).
UNIFG was involved in Task 1.1 for the identification of molecular markers to identify food borne pathogens such as Listeria monocytogens and E. coli O157:H7 in fresh cut vegetables. Firstly, a protocol for bacterial DNA extraction from vegetables samples, suitable for pathogen detection in PCR and Q-PCR assays was reported (deliverable D.1.1). Furthermore, protocol has been improved comparing several DNA extraction kits commercially available. A final DNA extraction kit was than chosen and used for all the experiments carried out. The occurrence of Listeria spp. and E. coli O157:H7 in minimally processed vegetables purchased from different retailers in Apulia region (Italy) was investigated, using both selective media, immunoassay tests and PCR detection tools. Moreover, MPN-QPCR methods for quantification of L. monocytogenes and E. coli O157:H7 from fresh cut vegetables and enrichment free methods for quantification of L. monocytogenes, E. coli O157:H7 and E. coli O104:H4 by Q-PCR were developed.
Primers were chosen among the ones available in literature in order to design QPCR assays that were highly sensitive and specific for purposes. A total of four QPCR assays were designed and tested for Listeria monocytogenes. Three of the primer pairs employed targeted virulence-related genes (iap, hlyA), while one set was used to amplify a sequence in the 16S rRNA gene. Two different chemistries, SYBR green (for 16S rRNA and hlyA(1) assays) and TaqMan (for hlyA(2) and iap), were used to set the QPCR assays. For E. coli O157:H7, primer sets targeting two different genes were tested during the study. Primers were selected from literature since are widely used in reference laboratories across Europe, for detection and enumeration of E. coli O157:H7. Both primer sets were coupled with fluorescently labelled probes in order to perform TaqMan assays. Target genes are rfbE, encoding antigen O157, and fliC H7, encoding flagellar antigen H7.
For E. coli O104:H4 primer sets and probes targeting genes wzxO104 and fliCH4 were tested during the study. These primers have been successfully employed for detection of E. coli O104:H4 during the large outbreak of 2011 in Germany. Both primer sets were coupled with fluorescently labelled probes in order to perform TaqMan assays.
For L. monocytogenes the MPN-Q-PCR method developed, can be used to shorten the MPN procedure from about 7 days to one day with a faster and reliable confirmation of L. monocytogenes in enriched samples (Fig. 2). The assay allows detection of L. monocytogenes down to 101 cells/g without enrichment and quantification of cells in concentration down to 102 cells/g after 6h enrichment.
For E. coli O157:H7, the MPN-Q-PCR method developed can be used to shorten the MPN procedure from 4 days to one day with a faster and reliable confirmation of E. coli O157:H7 in enriched samples. The assay is also suitable for rapid detection of pathogens as it allows detection of E. coli O157:H7 down to 101 cell/g after 2 hours of enrichment (See first period report). For E. coli O104:H4, the selected assays had proven to be highly specific and sensitive. Assays based on the oligonucleotides correctly identified E. coli O104:H4 in all of our tests while showing no signal in case of identified E. coli O157:H7 and non-STEC E. coli. The MPN-Q-PCR method developed was further confirmed in cooperation with the Istituto Zooprofilattico per la Puglia e Basilica - IZSPB (Foggia, Italy) on randomly selected fresh cut vegetables.
The aim of the research activities of the Task 1.2 performed by AUA were to assess the: (1) prevalence of L. monocytogenes and E. coli O157:H7 in ready to eat fresh products, (2) genotypic diversity of the respective isolates in terms of genotype, serotype and key gene content and (3) effect of storage conditions on the transcription rate of key genes at the ecosystem level.
Prevalence of L. monocytogenes was 7% in rocket, 6% in cucumbers, 6.6% in melon rinds, and 3.8% in strawberries. Prevalence of E. coli O157:H7 was 7% in rocket, 3% in cucumbers, 6.6% in melon surface and 3.8% in strawberries. Both pathogens were not detected in the edible part of the melon samples analyzed (prevalence 0%). Accurate estimation of prevalence could only be achieved by the parallel use of at least two chromogenic media; using only one chromogenic medium in fact leaded to over- or under- estimation of the prevalence depending on the medium. The ability to detect the pathogens in the enrichment broths through specific PCR and RT-qPCR was also examined concluding that plating chromogenic media with inhibitory agents against antagonistic microbiota cannot be reliably replaced by direct PCR or RT-qPCR in the enrichment broths.
Regarding the genotypic diversity of the isolates, all of them, even from the same sample or chromogenic media, were differentiated by genotypic profiling at the strain level. All isolates contained the necessary genes required for pathogenicity, i.e. iap, prfA, plcA, plcB, hly, actA, inlA, inlB, inlC and inlJ regarding L. monocytogenes and stx1, stx2, eae and ehxA regarding E. coli O157:H7, as detected by published specific-PCR protocols, with modifications due to possible small differences in the gene sequence.
Regarding the effect of storage temperature (4, 10 and 30 °C), product type (melon, rocket) and time (upon inoculation, 0.5 6.0 and 24 hours) on the transcription of key virulence genes of the pathogens, it was concluded that the transcription of hlyA, plcA, inlA, sigB, iap, plcB, inlB, inlC, inlJ, lmo2672 and lmo2470 at ecosystem level was indeed affected by the conditions under study. On the contrary, transcription of stx1, stx2, eae and ehxA were not affected by these factors.
UMIL was involved in the identification of molecular markers to identify microbial contaminants and to evaluate quality during the whole production chain. During the first year, the research group at UMIL in collaboration with UNIFG and AUA developed and optimized an ELISA method using specific antibodies against Escherichia coli and Listeria monocytogenes. Different buffers and strategies were tested in order to isolate the bacteria from inoculated fresh-cut vegetables. The protocol developed and optimised allowed the isolation and detection of the human pathogen bacteria in 22-24 hours. The method was applied on different fresh cut vegetables such as cucumber, melon and rocket. The validation of the method was carried out using samples collected on the market and during the demonstration period as suggested from the referees of the middle term project evaluation.
The isolation of putative molecular markers for quality evaluation was performed by transcriptome analysis of rocket plants exposed to different pre- and postharvest stresses. The sampling was performed after 24 h since the aim was to avoid the isolation of primary genes activation but those that might be really associated with quality losses. The transcriptome analysis yielded more than 30,000 transcripts and few of them were commonly expressed in all stresses. Since, the project was focused on fresh-cut the molecular markers were searched among those that were commonly upregulated in postharvest stresses. The identification of the putative genes as molecular markers was carried out considering their intensity of expression. The behavior of the most promising genes was studied during postharvest of rocket and melon. The putative molecular markers genes were correlated with physiological and biochemical parameters in order to be sure that they were associated with quality losses. The expression of these genes was studied in rocket and melon stored at 4 or 20 °C, simulating an optimized cold distribution chain and room temperature condition, which should speed up all the senescence processes. The genes that provided good results in both fresh-cut vegetables were used for antibodies production. In particular, the region of the genes that shown high conserved regions were chosen, because the idea was to identify molecular markers that can work for a wide range of vegetables. The antibodies were then used for the antibody array chip production. In particular, four antibodies were used for the antibody array production. The antibody array was tested as potential diagnostic tool for providing fast and reliable results on quality and safety of the fresh cut vegetable at any stage of the distribution chain. On the antibody array three antibodies against senescence-associated proteins, encoded from the genes identified from the transcriptome analysis, and one antibodies against E. coli were printed. The antibody array was tested on inoculated E. coli rocket samples and senescent rocket and melon samples. The results are very promising for bacteria detection and for quality assessment of fresh cut rocket. The use of this antibody array should give with one single analysis the overall quality of the fresh-cut produce.
S&T results on process control (WP2)
The aim of the activities carried out in WP2 was to provide useful tools for process control to the fresh-cut industry based on non-destructive and rapid measurements or evaluation, aimed to enhance the final quality of the product. In this WP partners UNIFG, Universidade Catolica Portuguesa (UCP), Cardiff University (CU), Nuvifruits and Markes International (Markes) were involved.
UNIFG worked on the prediction of the degradation of internal quality attributes of fresh-cut rocket salad and melons, starting from the degradation rate of external attributes (Task 2.1). This objective was reached in 3 steps: 1) obtaining the degradation kinetics of the most important quality attributes; 2) identifying relationships between the degradation of external and internal quality attributes; 3) validating the model. The first deliverable (2.6) consisted of two scales to objectively evaluate the appearance score of melon and rocket samples, which were also implemented in other partners’ activities such as CU/Markes, UCP/Nuvifruits and UMIL. The effect of temperature of storage on MA packaged products was modeled, by considering 3 storage temperatures. Data on melon and rocket showed that the “Weibullian-logistic” model was the best fit for the experimental data relating to the most important external and internal attributes (deliverable 2.7) compared to the traditional zero, first and second order kinetic models,. Moreover the model allowed the estimation of the shelf life both in isothermal and non-isothermal conditions. For the second step, degradation of external and internal quality attributes were related to each other by considering the storage time, t, as the only common variable (Fig. 3). A new mathematical equation able to estimate an internal quality attribute from an external one was obtained, also in this case in isothermal and in real condition simulations, highlighting its excellent ability to predict internal attributes (i.e. ascorbic acid content) through the easy measure of external indices such as appearance score, gas composition or color parameters.
Within Task 2.2 UCP focused at selection and implementation of efficient techniques to assess representative markers (chemical and/or biochemical markers) of nutritional and functional quality of ready-to-eat products with the final purpose of modelling and establishing the effect of different processing conditions on these relevant markers (Fig. 4). Fresh-cut melons and strawberries and ready-to-eat rocket leaves were fully characterized in terms of antioxidant capacity, and content of total antioxidant compounds (total phenolic content, total carotenoids, total anthocyanins). Qualitative and quantitative characterization of specific antioxidant compounds was also assessed to determine functional and nutritional quality. Specific conditions, namely package atmosphere and storage temperature were tested for ready-to-eat rocket leaves (rocket storage temperature experiments were carried out in collaboration also with CU/Markes) and fresh-cut melons, cut size for melon (in collaboration also with CU/Markes) and storage temperature for strawberries. All ready-to-eat products used in the experiments were supplied by NuviFruits and Estevão Luís Salvador.
Moreover, in Task 2.3 UCP together with Nuvifruits and Estevão Luís Salvador investigated the effects of the production process and storage on nutritional and phytochemical composition of melon, strawberries and rocket leaf products. Nutritional audits of processing lines of a fresh-cut fruit company and a ready-to-eat leafy vegetables enterprise were performed to generate recommendations for optimal preservation of phytochemicals of fruits and vegetables during processing through to the final product.
Audits were based on the enterprise’s definition of processing conditions (isolated or combined) of fruit and vegetables, their description in representative flowcharts, and identification of the process variables.
CU and Markes, performed an initial assessment of solid phase micro extraction (SPME) and thermal desorption, gas chromatography time-of-flight mass spectrometry (TD-GC-TOF-MS), and the latter was shown to be superior in robustness of collection method, sensitivity and lack of saturation (Task 2.4). This approach was therefore used to evaluate VOCs as a method for assessing effects of processing, storage time and temperature in melon and rocket salads, in collaboration with other QUAFETY partners. Following the mid-term review, the use of VOCs for assessing fresh cut strawberry quality was also evaluated.
In collaboration with UMIL and using scales developed by UNIFG, the complete VOC profile of fresh-cut melon cubes clearly differentiated samples stored at 4oC from those stored at 20oC, indicating that a breach in the cold chain would be detectable by this method. Several modules of compounds and selected single compounds were identified which look very promising for further development of VOC analysis for detection of storage temperature/ time evaluation. In collaboration with UCP, VOC profile analysis was shown to discriminate between different levels of melon processing. The discriminatory power was retained with the inclusion of only twelve compounds suggesting that the VOC analysis might be transferred to other platforms, and that by using TD-GC-TOF-MS, the analysis could be more targeted. VOCs could thus be used to assess changes in processing which may also be affecting nutritional value and or shelf-life.
In collaboration with UCP, the VOC profiles of rocket leaves were used to discriminate time x storage and showed good correlation to sensorial analyses and phytochemical content. VOC profiles also discriminated shorter term stress treatments of rocket leaves (in collaboration with UMIL) with a particularly strong effect of wounding, suggesting that VOCs may be of commercial use for assessing levels of mechanical damage during different processing pipelines.
Baseline data were obtained for fresh cut strawberries in line with recommendations at the mid-term review. A total of 63 compounds were detected of which 61 could be tentatively identified based on available databases, and the effect of storage time x temperature could be discriminated.
In collaboration with AUA, experiments on melon cubes inoculated with the human pathogen Listeria monocytogenes demonstrated that VOCs could be used to follow changes in levels of bacterial growth at different temperatures, and can detect levels of bacterial contamination. A 6 h period of incubation of the inoculated melon samples at 37oC increased the sensitivity of the VOC detection by 103 fold. This is still a short period of time compared to cultural or PCR based detection methods.
S&T results on decision support (WP3)
The aim of the research activities carried out in the WP3 was to provide tools for the industry operator to support decisions in very critical points of the fresh-cut processing chain from raw genotype selection to planning economic strategy for investments. In this WP partners Agricultural Research Organisation of Israel – The Volcani Centre (ARO), UCP, UNIFG, London Metropolitan University (LMET), Distretto Agroalimentare Regionale (DARe), Einat Food Industries Agricultural Cooperatives (EFI) and Nuvifruits were involved.
Within Task 3.1 ARO and EFI defined the criteria for choosing melon genotypes most suitable for fresh-cut processing and production of RTE melon products (an “ideal melon”). The performance of RTE products prepared from various melon genotypes was compared in storage trials. The study has shown that physiological behavior (climacteric vs. non-climacteric) affected product shelf life and off-flavour risk in response to packaging; rind surface (netted vs. smooth) affected the microbial attachment, decontamination efficacy and product safety; flesh color (orange flesh) reflected nutritional value associated with carotenoid phytonutrients; fruit size and shape affected the efficacy of manual and mechanical processing and waste amount; flesh texture and soluble solids level affecedt the product’s sensory quality. An inodorus-type melon cultivar Sorbeto was chosen as a model variety answering most criteria defined (Fig. 5). The performance of RTE Sorbeto melons was characterized in storage trials and further in demonstration activities (WP5) together with the industrial partner EFI and disseminated through WP8 activities. This genotype was used for developing RTE melon packaging (Task 3.4 WP3) and innovative mechanical processing (Task 4.5 WP4).
Moreover, in Task 3.4 ARO developed a method for optimizing modified-atmosphere (MA) packaging of RTE fruit in order to control their spoilage without a risk of oxygen depletion and off-flavour development. Microperforated packaging, with or without gas flushing (active MA) was used as a methodology to reach the aim. A user-friendly interactive decision-support tool predicting the in-package oxygen level as affected by package parameters (film type and thickness, perforation level, package dimensions, produce load, etc.) was built in order to adjust the packages for various commodities and conditions. The superior performance of the optimized MA packages was tested in storage trials with different melon cultivars (cantalupensis-type Raanan and inodorus-type Sorbeto) and further in demonstration activities (WP5) together with the industrial partner EFI. The results were disseminated through WP8 activities (Fig. 8).
UCP and NuviFruits performed the Task 3.2 related to the development of physiology-based models to assess the effect of oxygen partial pressure and storage temperature on ready-to-eat products respiration rate and quality parameters. Fresh-cut fruits and vegetables packaging trials were performed to allow modelling the degradation kinetics of quality changes and nutrient losses during post processing storage. Kinetic parameters found by linear regression for the orders 0, 1, and 2 allowed to select the most adequate kinetic order for the estimation of rocket leaves and fresh-cut melon quality changes under different modified atmospheres, and the goodness of fitting (Fig. 6). The best conditions found for the preservation of overall (physiological and biochemical) quality of ready-to eat rocket leaves and fresh-cut melons and strawberries were the initial package atmosphere of 5 % oxygen and the storage temperature of 5 ºC. Also, the functional description for respiration rate of ready-to-eat rocket leaves and fresh-cut melon, as function of oxygen partial pressure, was determined through O2 consumption levels monitored throughout refrigerated storage. A good fit of Michaelis-Menten model to experimental data was found and the kinetics of respiration as a function of O2 suggest that no significant reductions in respiration rate of fresh-cut cantaloupe can be achieved by lowering O2 levels. High R2 values for the Michaelis-Menten coefficients Vmax and Km were determined, although overestimation of O2 levels inside equilibrium modified atmosphere packages was common among fresh-cut produce tested. The results were obtained from WP2 parallel experimental trials and the results were disseminated through WP8 activities.
The Task 3.3 was performed by UNIFG in order to develop mathematical models to predict the barrier properties of polymeric films and the respiration rate of fresh-cut packaged products. The models can be used to provide information on the influence of the packaging characteristics (film thickness, presence of micro-perforations, etc.) on the respiration rate of fresh-cut packaged products (Fig. 7). This will make it possible to predict how to package a particular type of product only from the knowledge of its respiratory rate. For the first aim, monolayer, multilayer and micro-perforated films with different hole diameters as well as variable distance between them were taken into account, being different in terms of barrier properties. For the model validation a comparison between predictive and experimental data was carried out. In order to quantitatively determine the model prediction goodness, the mean relative deviation modulus was calculated. The model used to describe the packaged food respiration rate was obtained by writing the mass balance equation of gas in the package headspace of fresh-cut melon. The developed models are of great interest in packaging design of fresh-cut produce.
LMET studied the effects of mild (sublethal) heat treatment, growth at sub-optimal pH values (sublethal acid stress) and subsequent recovery temperature on the lag times of individual cells of Listeria monocytogenes and Escherichia coli (Task 3.5). It was found that, except for acid stressed L. monocytogenes, which showed a slightly short mean value for individual lag time at 10 oC compared with that at 22 oC, the mean and standard deviation (SD) of individual lag times of L. monocytogenes and E. coli increased when recovery temperature decreased. The mean and SD of individual lag times of L. monocytogenes increased after sublethal heat stress, while E. coli individual cells showed poor recovery at 10 and 15 oC after heat stress. Acid stress resulted in an increase of individual lag time for L. monocytogenes at 5 oC, but, surprisingly, in a decrease at 10 oC and no significant change at 22 oC, compared with undamaged cells. For E. coli, acid stress appeared to result in a decrease in individual lag at 10, 15 and 22 o C compared with the undamaged culture. The zero adjusted Gamma distribution was demonstrated to be the best fit to data for individual lag of both L. monocytogenes and E. coli. A quadratic polynomial relationship was developed to link the parameters of the zero adjusted Gamma distribution of individual lag with the recovery temperature. The performance of the model was validated by comparing the population lag of undamaged, heat stressed and acid stressed L.monocytogenes and E.coli in melon with values predicted by the model for the same environmental conditions. Except for the heat stressed model for L. monocytogenes, which provided a prediction for lag slightly comparable to the experimental population lag in melon, it was found that in most cases, the predicted population lag was significantly shorter than the observed lag in melon. Although the model “fails safe”, this underestimation of lag highlights the need for more conditions to be tested to improve performance of the model. In addition, L. monocytogenes cells subjected to combined acid and heat stress may pose a risk to consumers of fresh cut foods, such as melon, at the end of two weeks’ refrigerated shelf-life. Recovery of individual cells of E. coli after application of combined acid and heat stress was significantly affected by subsequent incubation temperature (Fig. 9).
In the Task 3.6 DARe conducted focus groups and in depth interviews among partners with the aim of identifying the innovations produced within the project, their impact on production and their requirements for the implementation. The outcome identified the key information for a direct communication between companies and QUAFETY research for a real understanding of the impact of the project on products or firms performance indicators. Having organized systematically the information DARe set up an excel spreadsheet based software providing a decisional tool to industry that will support innovation investments. The software has been, then, tested on a sample of 17 companies, which provided their feedbacks. The main outcome of the feedbacks concerns the possibility to better understand innovations and to furnish a more easy connection between innovations, their impacts on production, their requirements, and contact with researchers. In such a way, companies declared that with high probability will contact the researchers in order to have further information on the implementation in their production process (Fig. 10).
S&T results on innovative processes (WP4)
The aim of the research activities carried out in the WP4 was to develop and/or to evaluate the application of innovative processes to improve quality and safety of the final product. Looking at the whole farm-to-fork chain, the first step to consider is growing system and pre-harvest practices management. Partners involved in WP were ARO, UNIFG, the Instytut Ogrodnictwa (INSAD), EFI, and Agronomia Scarl (AGRONOMIA).
The purpose of the work performed by UNIFG in the Task 4.1 was to obtain raw material from several leafy vegetable species with improved characteristics in terms of safety, nutritional quality and shelf-life. The purpose was reached by strategically managing the nutrient solutions in different soilless cultivation systems. The research activity was focused on the definition of innovative optimized growing protocols for several model species including lettuce, endive, escarole and rocket salad (Fig. 11). The innovative protocols were based on soilless cultivation systems, considered the most intensive production method in today’s horticulture industry, using electrical conductivity levels of nutrient solutions from 1.5 to 4.5 dS m-1. Two cultivation systems were tested: a modified Ebb and Flow system and a Floating System (FS). Strategies aimed at the abatement of nitrate content in the leaves were also addressed. The following main results were obtained:
1. The best electrical conductivity of nutrient solution was 3.5 dS m-1. This level provides the best yield and quality performances giving leaves with low nitrate, high antioxidant compounds and high dry matter content as well as high shelf life;
2. The modified Ebb and Flow system appears more suitable with ‘lollo’ lettuce, while the Floating System performed the best with rocket salad, endive and escarole;
3. Nitrate content can be successfully lowered by substituting NS with water, but the strategy must be used no longer than the last 3 days before harvest;
4. The quality of soilless-grown rocket salad depended on season. The product grown at warmer conditions had better yield, visual quality, dry matter and vitamin C and keeping quality.
In order to reach the objectives of Task 4.2 AGRONOMIA tested the performance of water ultrafiltration system for pre-treatment, treatment and cleaning (disinfection) of water under the conditions of a commercial fresh-cut plant specialized on leafy vegetables processing. The ultrafiltration technology was best suited for the treatment of the process water, compared to quartz and metal mesh filters (Fig. 12). The ultrafiltration technique permitted to re-use the water in the process and to increase the volume of clean water used at the washing step. As an additional effect the system allowed to recover some of the water used for washing, and to put it back on circulation reducing the environmental impact of the whole system. In order to prevent trihalomethanes accumulation in ultrafiltration systems the hypochlorite was substituted with 2.5 ppm of chlorine dioxide sanitizer. The advantages of chlorine dioxide are its efficacy at low concentration, ease of use and lack of toxic byproduct residues. Combination of ultrafiltration with chlorine dioxide represents a significant alternative to chlorine in washing water sanitation of fresh-cut produce.
ARO (in cooperation with EFI) tested the efficacy of rind decontamination of melons intended for fresh-cut processing in order to ensure microbiological safety and to control spoilage microorganisms on ready-to-eat products (Task 4.3). A new method for thermal decontamination of melons and a prototype installation for realization of this method were designed and evaluated. The melon surface temperature reached 120 to 200 °C after 30 sec of the thermal decontamination. The treatment allowed the reduction of microbial population on the melon surface by 4.5 to 6 log CFU cm-2 while the efficacy of the standard hypochlorite treatment did not exceed 3.5 log CFU cm-2 (Fig. 13). After 30 s treatment, the heating was restricted to melon rind (less than 2 cm depth) and did not affect the flesh temperature. No negative effect on the fresh-cut product quality was observed. The optimal treatment duration was 20 to 30 seconds. The work has demonstrated the feasibility and efficacy of the proposed approach for decontamination of melons intended for fresh-cut processing.
Moreover, in Task 4.5 ARO improved the efficacy and safety of fresh-cut melon and watermelon processing by substituting manual peeling and cutting operations by an automatic hands-off machine. The study was designed to develop a technology and machinery able to mechanically cut and peel melons and watermelons in order to prepare fresh-cut, ready-to-eat products (Fig. 15). As part of this study several peeling alternatives ranging from laser-cut to high-pressure water using artificial vision systems, as utilized in sorting processes in the production industry, were compared. The study evaluated, analyzed and developed a number of special puncturing tools: fixed, rotating, solid and perforated. It was found that a round, disk shape slice as final product is achievable. The results of these trials have led to the design of the system performing peeling/cutting operations without a human hand touching the cut fruit. At this stage the machine requires a manual feed of the raw fruit. The main parts of the machinery have been built and tested successfully. The mechanically excised products were found to maintain the acceptable quality for more than a week when stored in sealed plastic packages in cooled environment.
Within Task 4.4 UNIFG analyzed the main critical factors affecting the formation of Listeria monocytogenes biofilms in the fresh-cut industry and to find the efficient approaches to its control (Fig. 14). The work strategy included three parts: (a) biofilm development in Listeria monocytogenes EGD-e and role of the agrA gene in biofilm formation; (b) biofilm formation in Listeria monocytogenes of vegetables origin; (c) inhibition and removal of established biofilms by L. monocytogenes using different oxidizing agents. The following significant results were obtained:
1. Temperature is a critical factor for biofilm formation by L. monocytogenes. Low temperatures should be maintained in the work environments in order to minimize the biofilm formation.
2. Listeria monocytogenes strains from vegetables origin are able to form biofilms. Biofilm formation is variable between strains.
3. Biofilms may be formed independently from the surfaces (glass, polystyrene and stainless steel) although hydrophobic materials should be preferred when compared to hydrophilic supports.
4. Commercial biocides may effectively remove biofilms formed by L. monocytogenes of vegetables origin. Sodium hypochlorite was effective when applied after the incubation of L. monocytogenes. In contrast, benzalkonium chloride and peroxyacetic acid were efficient only when applied before incubation of L. monocytogenes strains.
The activities carried out by INSAD in Task 4.6 were aimed to test a novel postharvest hot-water treatment as a mean for improving quality and safety of fresh-cut vegetables and extending their shelf life. During the reported period the research was carried out on Chinese cabbage, rocket leaves (salad and wild type) and melon (Galia and Cantaloupe type). The storability of Chinese cabbage cv. Bilko F1 was positively affected by brief (3–5 s) dipping in hot water at 53–55°C (Fig. 16). The hot water treatment delayed browning of cut cabbage leaves surface allowing to maintain better quality during storage. Sensory analysis performed after short term storage confirmed the superior sensory characteristics of cut Chinese cabbage dipped in hot water (53–55°C) as compared with non-treated control. Hot water treatments generally had no significant effect on chemical composition and microbial population of the fresh cut Chinese cabbage.
In contrast to Chinese cabbage, only limited quality improvement was reached with rocket and melons. With rocket, brief dipping (3–5 s) in hot water at temperature 50-55°C retarded the leaf yellowing during short term storage. However, the treatment was associated with the risk of heat injury.
The activities of Task 4.7 were performed by AGRONOMIA in cooperation with ARO to examine the efficacy of ultraviolet (UV) light for improving quality and safety of fresh cut products. UV treatment had a positive effect of reducing the microbial population on ready to use leafy vegetables. The greatest microbial inhibition was obtained using long time of exposure (more than 10 minutes, 8 kJ/m2) which caused a too high stress to the delicate rocket leaves. The smaller UV doses (2 or 4 kJ/m2) still had a positive effect of reducing the microbial population and at the same time exerted less phytoxic damage. With endive, the problem of the UV damage was far less acute than with rocket. The UV treatment resulted in certain increase of phenolic compounds content in rocket leaves and endive. The UV treatment proved itself an efficient instrument to inhibit the microbial proliferation during shelf-life of fresh-cut melons (Fig. 17). The treatment had no negative effects on the quality and nutritional value of fresh-cut melons and induced even a slight increase in the antioxidant activity. In the production of fresh-cut melons the UV treatment can be applied without hampering the processing chain when the fruit cubes are filled into open retail containers and before these containers are sealed.
In Task 4.8 , AUA, UNIFG and ARO worked together in order to develop active and intelligent packaging solutions for use with fresh-cut produce. The active packaging was based on active antimicrobial compounds using two methods of release to the product surface: (i) from the packaging through the hedspace and (ii) via edible coating. Regarding the intelligent packaging, the work aimed to develop a novel Time-Temperature Indicator of the product freshness. The following major results were achieved. (1) The vapors of lemongrass essential oil released by the packaging material affected the enterococci and yeast and mould populations depending on product, storage temperature and packaging atmosphere. The impact of the essential oil on product’s sensorial properties was pronounced in first 4-5 days of storage and faded as storage proceeded, due to the oil evaporation (Fig. 18). (2) T-gelatin films prepared by the Layer-by-Layer approach demonstrated good mechanical properties, higher permeability and effective microbial spoilage inhibition. The Layer-by-Layer (LbL) formulation was superior compared with monolayer formulations in preservation of fresh-cut melon texture and slightly reduced fruit weight loss. Both LbL and blended composite coatings did not obstruct fruit gas exchange and did not cause accumulation of off flavor volatiles. (3) The best combination of process variables corresponded to proper washing of the whole melon, dipping treatment of the cut pieces in hexanal, active coating with hexanal and packaging in OPP40 (Fig. 19). Under these conditions the shelf-life reached 16 days, compared to less than one week in the control. (4) A Time-Temperature Indicator based on physiological activity of lactic acid bacteria was developed and successfully tested with rocket leaves storage; its presentation on the packaging was proposed.
In Task 4.9 the work performed by AGRONOMIA as first step defined the specifications to fabricate a prototype of Passive Refrigeration System (PRS) container that could be used in the open field for collection of baby leaves or whole salads, for refrigerated transport, for short storage during processing, and for final distribution, maintaining throughout the chain the temperature between 0 and 4 °C. The seconf step was to test the PRS performance in the presence of product. The device confirmed its capacity to cool the product. The baby rocket leaves were found more prone to low temperature damage than the crisphead lettuce. The eutectic plate preparation temperature had a major effect on the efficacy of the PRS system (Fig. 20). With eutectic plate frozen at -15°C (completely frozen) the temperature in the PRS box dropped below the conventional forced-air transport conditions that might cause damage to rocket at pre-processing stage and to crisphead lettuce during shelf life. Eutectic plate chilled at 0°C (therefore not frozed) did not allow reaching appropriate temperatures, so the product encountered microbiological problems during storage, reducing the shelf-life in both commodities. Eutectic plate frozen at -4°C (partially frozen) allowed reaching appropriate temperatures in the PRS box comparable with the conventional forced-air refrigeration so that the products preserved similar or improved organoleptic and microbiological quality compared to traditional method. The limited quality improvement by PRS container compared with the traditional standard forced-air system was due to the brief time of transport (max 18-20 hours). Using PRS containers for longer distribution distances would maximize their benefits.
S&T results on demonstration activities (WP5)
Demonstration activities were aimed to prove the technical viability of the new technologies developed in the project, thus boosting at industrial level the adoption of new expertise and innovative tools able to ensure safety and quality of ready-to-eat fresh produce throughout the whole chain. The industrial implementation of research findings was clearly stated and planned including process optimization and management procedures. The expected interactions among partners inside and outside the consortium were widely respected, thus giving a multidisciplinary approach (Fig. 21). The activity was related to the results obtained within the first four WPs of the project, by interfacing diagnostic kit (WP1), process control (WP2), decision support (WP3), innovative processes and management system implementations (WP4). Therefore, WP5 was articulated in four tasks, from Task 5.1 to Task 5.4. In particular, in Task 5.1 involving UNIFG, AUA, and UMIL with AGRONOMIA and Nuvifruits, as partners and the Institute Zooprofilattico per la Puglia e Basilicata (ISZPB) as public Health Institution. The demonstration activity was aimed to give to the SMEs the opportunity to innovate their quality management. The minimal instrument and tools were tested for making as easier as possible the use of the protein antibody arrays for quality evaluation and microbial contamination. The demonstration was focused on the bacteria detection (Listeria monocytogenes and E. coli) using an ELISA based method developed in the WP1. This method compared with the standard procedures allowed gaining detection time less than 24 h, while the standard methods take 4-5 days. The implementation of the developed diagnostic kits allowed checking the quality of dubious fresh-cut vegetables, before the transporation of the final product, in order to guarantee the consumers safety and quality of the products. In addition the application of a quantitative Real-Time-PCR (Q-rtPCR) method associated to MPN for pathogen detection and enumeration in fresh-cut products was also demonstrated with the cooperation of ISZPB. Task 5.2 was aimed to assess the efficacy of process control by the involvement of UNIFG, CU and UCP with AGRONOMIA, Nuvifruits and am additional company external to the project. Quality attributes of rocket leaves and melons were monitored during distribution by temperature data loggers, so to have a thermal history of the products. Starting from the external attributes, internal attributes were predicted by applying the models, and then verified through the application of standard destructive methodologies. Analysis methods for remote collection of volatiles were also demonstrated. The evaluation of volatile profiles associated with deterioration under different storage conditions in relation to appearance, progression of degradation in terms of growth of spoilage organisms, nutritional and organoleptic quality was demonstrated with the objective to complement and expand the destructive diagnostic methods. The VOC demonstration used TD-GC-TOF-MS for assessing quality of fresh-cut rocket salad and melon at the start of processing and during shelf life of the processed products. Task 5.3 related to demonstration of decision support involved Unifg, UCP, ARO, LMET and DARe with Nuvifruits, AGRONOMIA and EFI as industrial partners. Models to assess the benefits of MAP on fresh-cut fruit and vegetables and models to predict the barrier properties of packaging developed in WP3 were validated by using melons from Nuvifruits and rocket salad from AGRONOMIA. In both cases, different steps were carried out and in each of them the main quality parameters were assessed. As regard to model-driven control of spoilage and off-flavor in RTE fresh-cut fruit products, the efficacy of micro-perforated active modified atmosphere (MAMA) packaging for preservation of fresh-cut melons was demonstrated with success at the commercial fresh-cut facility of EFI. The activity of LMET was aimed to determine single cell lag phase durations of pathogenic organisms, e.g. Escherichia coli and Listeria monocytogenes, and to apply statistical methods to distributions of single cell lag times in order to develop models that more accurately predict lag phase in fresh-cut products. The developed models were demonstrated to be suitable tools for decision-making in the food industry. The application of a multi attribute model for investment decisional support through an user-friendly software, developed in Task 5.3 was carried out by direct demonstration through DARe personnel and indirect demonstration through an online manual. In this activity industrial partners within QUAFETY and other SMEs were involved. The main result was the application of the software in investment decision routine, use of the software for analyzing farms issue beyond financial aspects for investments and inclusion of QUAFETY know-how into farms. The activity of Task 5.4 was aimed to demonstrate the innovative processes developed by UNIFG, AUA, INSAD and ARO to the industrial sector by involving EFI and AGRONOMIA as project partners and Farm De Ceglia as external company. The efficacy of studied solutions to increase quality and safety of fresh-cut products was demonstrated. Particularly, benefits of soilless cultivation protocol applied on leafy vegetables were underlined, (raw material with high quality and safety, with higher shelf-life, with a better esthetic value and with a better nutritional value), thus validating under real farm-scale conditions the results obtained in the experimental-scale. The effectiveness of developed waste-water sanitization system as chlorine-alternative method was assessed in the company AGRONOMIA. The efficacy of UV treatment of fresh-cut melons microbial load was demonstrated at commercial fresh-cut facility of EFI. The partners involved were ARO and EFI. The effectiveness of biofilm inhibitors and biofilm removing agents on Listeria monocytogenes was demonstrated by UNIFG. Demonstration was carried out by using biocide on L. monocytogenes able to form biofilm on industrial surfaces, to give a protocol related to “the best practices” to be followed in order to reduce biofilm formation during fresh-cut production and risk of biofilm harbouring L. monocytogenes. The partner INSAD was involved in demonstration of hot water treatment applied to rocket to prolong the shelf life. In the same task the suitability of developed active and intelligent packaging applied to rocket salad was assessed based on quality and safety criteria.
S&T results on Evaluation of economic viability and sustainability of the applied methodologies (WP6)
WP6 focused on testing the hypotheses of ready to eat fruits and vegetables as substitutes for conventional whole products, and quality attributes deriving from innovative production technologies, within those produced by QUAFETY research, in order to understand consumer preferences and draw consideration on the consequent consumer acceptability and measuring consumer perception towards these attributes. In addition, consumers’ acceptability measures were used for setting up the cost-benefit analysis of the innovations produced by QUAFETY. In this WP all partners were involved.
This WP was divided into three tasks. Task 6.1 (first year activities) includes all the activities leading to the estimation fresh cut mathematical functions of consumer preferences, allowing the prediction of consumption behavior towards RTE fruit and vegetables at different levels of price/quality/convenience. This result allows inferring on consumer preferences, consumer perception and provide information to orient applied research and investors choices. Task 6.2 (second year activities) consisted into estimating willingness to pay for different quality and convenience levels. Finally, Task 6.3 (third year of activities) focused on the assessment of the impact on costs and revenues of the new technologies coming out from QUAFETY project.
In order to depict the quality attributes enhanced by the innovations produced within the project, from a consumption perspective, in depth interviews have been carried out within the Task 6.1. QUAFETY Partners have been involved in a survey in which specific and in-depth questions have been asked in order to contextualize the outcomes of their project research in a future real product (concept), in which those innovations are embedded into it, and perceived by consumers. Once answers from research partners have been collected, data have been analyzed and discussed in a focus group environment among researchers of D.A.Re. The discussion, aimed to translate research products into quality attributes, labels, price and shelf life variations, generated the list of attributes and levels to be analyzed in order to build a mathematical function that predicts consumer choices towards RTE products.
Having the attributes, an experimental design was built in order to propose a limited number of alternative products to consumers during the subsequent survey. The alternatives represented “real products” together with “conceptual products” from which consumers were asked to choose.
Finally the survey was conducted with a total of 1,462 interviews across some EU Countries. Cooperation among partners has been fundamental for the implementation of the survey and translation/interpretation in local language, given that fresh cut market has its own specificities in each country. More specifically, about 500 consumers were interviewed in Italy, about 250 in Spain, about 250 in Greece and about 500 in UK. Consumers were selected randomly at the exit of the shopping mall across the entire area of the countries (Italy, Greece) or while shopping online (Spain, UK). However, the sample was stratified by age and gender in order to increase the level of representativeness.
Once data were collected an electronic database was input with all the information. Sample descriptive statistics was prepared and the preference predictive model was set up in order to understand consumer perception of different attributes, and to forecast purchase probability. In order to select the best model, several alternative models were tested competitively.
Moreover, cross-country comparison was conducted in order to depict the main differences in habit, preferences and attitudes toward consumption.
In the Task 6.2 the predictive preference model has been tested and improved in order to calculate the best price at which consumers show the strongest preferences. The hypotheses in the WP test whether quality attributes deriving from newer production technologies have impact on consumer acceptability, hence their willingness to pay towards each novel product attribute (Fig. 23). Knowing the impact on consumer willingness to pay of each innovation helped hypothesizing prioritization of innovation based on profitability of the product differentiation obtained by QUAFETY research. As a consequence, the willingness to pay was modelled and calculated, based on consumer survey data and preferences analysis. With WTP measures, based on empirical models of consumer choice prediction, and set up on survey data, consumer response to the quality attributes deriving from new technologies could be measured in order to evaluate the impact on the consumer price responses towards the innovations analyzed in the project. The analysis of the willingness to pay values was also focused on acceptability measures, in order to be sure that selected attributes did not represent an attended attribute, hence did not orient preferences and choices. Finally, cluster analysis has been conducted in order to better identify eventual marketing directions. We found that shelf-life extension did not generate positive consumers’ reactions as well positive willingness to pay, as opposed to safety enhancement and additional information to consumers.
Activity of the Task 6.3 focused on the cost-benefit analysis, which took into consideration results of willingness to pay calculation, together with the results of a questionnaire to researchers, which involved the collaboration of all project members, in order to calculate cost information relative to QUAFETY innovations and measures the level of profitability for each innovation produced within the project. Cooperation has been fundamental for the individualization of the impact on costs of each innovation produced. Therefore, the activities aimed to estimate the impact of innovations on final company profit and furnish economic viability measures to assess the mere business validity of the innovations produced within QUAFETY research. Results have been considered starting from the perspective of two extreme types of representative companies, from cost structure point of view, and consider different price perspective in the medium long run. For instance, there are innovations affecting positively the reduction of spoilage costs, but an increase in depreciation, because they need more fixed investments. Other innovations increase costs relative to personnel training but increase revenues, etc. Meanwhile, when analysing the implementation of innovations, fixed investments are also taken into consideration. Fixed investments, in fact, require an initial very high cost, with a consequent yearly depreciation. The cost-benefit analysis considers also how eventual increase in profits pays this investment back. As a result, many of the investments proposed by the QUAFETY innovations did not get high returns of substantial increases in the net present value of the company. As a consequence, fixed investments are not always desirable. In those cases, researchers should consider the provision of third party service, which will not need the implementation of fixed investments in the company, but will only increase variable costs of a given amount.
Finally, DARe results, obtained with the full collaboration of all partners, allowed to state that not all innovations impacted positively on the willingness to pay and not all innovations generated a gain in total profits. There are innovations that do not impact price positively nor reduce costs, making production more efficient. In those cases, the probability that companies will adopt the innovation is noticeably reduced, while other innovations could effectively contribute to a profit increase, and be adopted for convenience reasons. Thus, the indubitably positive impact of knowledge and quality of research does not always get positive response in terms of company convenience. However, obtained results provided a critical approach to look at industrial scale-up and implementation of research results for profit oriented reasons. Nonetheless, the revision of some of the innovations, with a managerial perspective, could lead to higher level of convenience.
S&T results on Safety & Quality Management System (WP7)
WP 7 was organized into three tasks to develop assessment tools for quality and safety of the SME of the fresh cut sector, to apply and analyse the collected data and finally to propose potential interventions for improvement in the participating SMEs. To achieve these objectives all partners actively participated to the above tasks. The main results of the WP7 were the development of a diagnostic instrument for the assessment of the management systems applied in fresh cut sector related to food quality and safety which include a questionnaire and a safety assessment plan (SAP). The questionnaire consists of five sections. The first includes background information for the SMEs, the second and third information on the risk factors, the fourth related to the QMS/FSMS applied in SMEs, if any, and the last one includes questions for good agricultural practices during pre harvest, prerequisite programmes and processing control points. The responses from the application of the questionnaire followed the process described in Fig. 24. Thus, applying principal component analysis significant components were extracted to provide a score for QMS/FSMS.
The developed SAP was aimed to assess the level of safety along the production-processing chain and the safety level of the products. In this assessment plan, the sampling points were identified, the required number of samples were determined, and the appropriate analytical methods for the sampling and determination of safety and quality level at the different sampling points were indicated. After the application of SAP, the data were collected and analyzed. This SAP will be a key part of the food safety and quality management system that will be used in combination with the questionnaire for the assessment of the overall performance of the QMS/FSMS of the participating SMEs and to draft the recommendations for the improvement of the management systems applied in the SMEs. These will be an integral part of the management systems applied (Fig. 25).
The assessment of the QMS/FSMS revealed a disagreement between the scores from the questionnaire and the scores from the interpretation of the SAP in 2 out of 4 SMEs (Fig. 26). For two SMes very good agreement between the two parts of the diagnostic instrument (questionnaire and SAP) was found: SME C ranked at the “top” level for both parts of the instrument, whereas SME A ranked at the “good” level for both parts of the instrument. On the contrary, disagreement between the two parts of the instrument was observed for the other two SMEs: SME D and SME B were ranked at the “top” level according to the questionnaire and at the “moderate” level according to the interpretation of the SAP. In cases of disagreement between the questionnaire and the SAP, the latter provides more objective evidence of the safety and microbiological quality status of the SME. Very often a bias is inserted in an attempt to ameliorate the real safety status of the SME. In such cases, a detailed auditing will give the opportunity to depict the actual safety status; at the same time training of the personnel is necessary in order to understand the importance of accurately answering in such audits.
The impact on other WPs is defined by incorporation of these tools in the FSMS guide of WP8 and to decide which of the procedures developed in WP1 to WP5 will be useful alternatives to improve the processes used in the fresh cut produce sector. The creation of a FSMS guide is the result of info and data collected by all partners trough the experimental work and demonstration activity in the SMEs. This guide should be a tool for daily use; these guidelines should help fresh-cut industry operators to manage and improve safety and quality of the fresh-cut products. The guide is available in USB-key format for fresh-cut chain operators.
Dissemination (WP8)
The Objective of WP8 was to disseminate project results to potential users (i.e. Food industries, consumers, health institutes, mass media, etc.) by online websites, seminars, training courses, conference attendance, papers, etc. The dissemination activity is the result of a great and continuous cooperation among all Quafety partners.
The objective of Tasks 8.1 and 8.2 consisted in creation of a subsection dedicated to fresh-cut management on all 3 Freshplaza’s websites (English, Spanish, Italian); in this subsection, called Quafety, divulgative articles on fresh-cut produce sector are published to disseminate scientific studies and to create a connection among the play actors of the whole fresh-cut chain. Furthermore, the official website of the project (www.quafety.eu) has been created by Freshplaza, while it is managed by all partners under leader supervision.
For dissemination work, more than 500 articles (Task 8.3) on fresh-cut produce chain were published and are online available at:
1. http://www.freshplaza.com/sector/109/quafety
2. http://www.freshplaza.es/sector/101/Quafety
3. http://www.freshplaza.it/sector/101/Quafety
The dissemination of scientific results, technological news, educational events and training courses of QUAFETY and fresh-cut produce chain will continue also behind the QUAFETY conclusion.
A guide on Food Quality Management System (FQMS) for fresh-cut segment was created and is available in USB-key format (Task 8.4). The guide is a list of 1) good agricultural practices (GAPs), 2) good hygiene practices (GHPs), 3) good post-harvest management practices (GMPs), 4) good processing practices for fresh-cut industry, which are crucial for HACCP (Hazard Analysis and Critical Control Points) system. The guide is the result of info and data collected by all Quafety partners through the experimental work and demonstration activity in the cooperating fresh produce companies.
Freshplaza created ‘QUAFETY group’ on Tacler (www.tacler.com) which is a Fresh Produce (fresh fruit and vegetables) industry web-based community platform. This platform was used as a permanent online network for connecting stakeholders with Quafety scientists (Task 8.5). The permanent interaction between scientific community and industry is in fact crucial for increasing company competitiveness in the global market. Tacler users from all over the world, daily demonstrate their need to enter new markets, to improve their products in terms of quality, safety and added value, to be competitive of economical and environmental sustainability. In Tacler, Freshplaza organized meetings with QUAFETY scientists to create connection with fresh produce chain actors registered to Tacler. This initiative represented the starting point of this new Tacler joint mission, with the idea to continue in the future this type of meetings.
Through dissemination activity, Freshplaza divulgated all info on training and educational courses, conferences and event for moving students, scientists, and fresh-cut chain operators (Task 8.6).
At the project conclusion, Freshplaza organized the final workshop in Rotterdam on 25th March 2015 (Task 8.7). During the workshop all QUAFETY results were showed and a CD-Rom with the most important project documents was created to distribute to the participants.
Potential Impact:
Project QUAFETY has proposed a complex program of food quality & safety solutions throughout the farm-to-fork supply chain for RTE fresh produce. Adoption of these solutions by SME producers may potentially improve their products and thus strengthen their competitiveness in the market. Furthermore, the innovations introduced by QUAFETY may improve sustainability of the technological process by substituting the use of hazardous and environment-polluting chemical sanitizers.
As the category of RTE products is clearly consumer-oriented, all solutions proposed by QUAFETY also address the needs of consumers that due to time limits or other circumstances choose not to prepare meals “from scratch”. RTE products are intended for direct consumption and so the acceptance of these products by consumers depends on their trust in measures undertaken by producers to ensure the product safety and quality. An increased level of confidence will go in line with the task of promoting fruit and vegetable consumption in Europe in order to reduce the risk of obesity and chronic diseases (European Parliament resolution 2010/C 8 E/18).
In addition QUAFETY results provided scientific evidences which may be exploited by health authorities (at EU, national, and regional level) in order to evaluate whether further regulation is required in this area of food safety and to support educational campaigns for healthy nutrition.
Another most important impact of the Project is associated to its function as a reference point for specialized and general media when covering quality and safety aspects of RTE fresh produce. As in the last few years fresh-cut products have been the target of a number of media attacks the presence of QUAFETY evidences has contributed to a more equilibrated approach to the topic of food safety.
Finally, last but not least, as a large of information coming out from a R&D project like QUAFETY are basic scientific information (representing major or minor breakthrough in the knowledge on specific topics), it has largely impacted with the scientific community throughout a number of scientific communications either presented in Conferences or published on peer-reviewed Journals.
Main “impacting” aspects of the Project are listed below by workpackage, while a summary of main dissemination numbers will be described thereafter.
Potential impact for WP1
The results obtained on the MPN-QPCR methods for quantification of L. monocytogenes and E. coli O157:H7 from fresh cut vegetables and enrichment free methods can enhance the safety of the whole fresh cut market.
Pathogen prevalence in fresh cut fruits and vegetables is essential for risk assessment protocols. This information along with the measure of genotypic diversity should be taken into consideration when designing intervention strategies aiming at the safety improvement of the fresh cut sector. At the same time information on gene transcription may improve our understanding of the pathogens physiology that may contribute to the development of more efficient tools for their control.
The antibody array containing specific antibodies against the senescence associated proteins and against the human pathogen bacteria allow the evaluation of quality and safety of the produce in fast and reliable way, at any step of the whole distribution chain from the grower to the consumer. The idea is that with a simple sample homogenization the crude protein extract can be used to hybridize the antibody arrays and then with a scanner highlight the presence of protein associated with quality losses or the presence of harmful bacteria (deliverable D1.3 UMIL). This diagnostic kit can enhance the quality and the safety of the whole fresh cut market.
Potential impact for WP2
The results of Task 2.1 provided useful applications at different levels of the research. The photographic scale developed in Deliverable 2.6 provides an objective tool for the evaluation of appearance of rocket and fresh-cut melons, which can be used for the evaluation of raw material both in research and by companies. The degradation kinetics in packaged samples both in isothermal and non-isothermal conditions were obtained, as reported in Deliverable 2.7. The kinetics may help processors to understand the most critical factors for shelf-life and specifically the impact of temperature. As complementary results the objective of an accurate shelf-life prediction, taking also into account the fate of nutritional components, was achieved. Finally the models relating changes of internal nutrients to appearance degradation may be a useful tool for having information on nutritional content, without the use of sophisticated methods and instruments.
Results obtained in Tasks 2.2. and 2.3 allow the definition of product shelf-life concerning their bioactive properties and protective health effects. Also, the studies undertaken in these tasks allowed the development of an expert-system available for the industry to predict the depreciation of the functional properties over time under different processing conditions. (http://www.quafety.eu/index.asp?partner=1&news=171). The “Expert System” program gathers, transfers and gives worldwide access to the knowledge obtained throughout QUAFETY Project on nutritional quality of processed rocket leaves, melons and strawberries as affected by processing (production process; cutting degree) and storage conditions (package type, modified atmosphere and storage temperature). It thus provides the industry with useful information for developing processes and making management decisions on the modulation of nutritional quality and shelf-life extension.
Task 2.4 generated useful baseline data on VOC profiles for melon and rocket (Deliverable 2.11) as well as for strawberries. It also identified potential markers based on VOC analysis of use to the processing industry for assessing processes which affect shelf-life such as mechanical damage and changes in temperature control as well as the quality of incoming material for processing (Deliverable 2.12). Interactions between CU/Markes and UK processing companies indicate a strong interest in taking this forward. The interaction with UCP also linked the VOC markers to sensorial data and analyses of compounds of nutritional significance thus VOCs could also be used to assess changes in processing which may also be affecting nutritional value and or shelf-life. The potential of VOC analyses for detection of contamination with human pathogens would also be of benefit to the industry as a complementary approach to other methods that are able to detect very low levels of contamination more rapidly that the culturing methods currently used.
Potential impact for WP3
The results of Task 3.1 provided a decision-support information for operators (growers and processors) choosing genetic material for producing high-quality RTE melon products. It will guide breeders in creating new melon genotypes for fresh-cut processing. For consumers, knowing a genetic origin of RTE melon may indicate a chance of off-flavour development during storage.
The kinetics determined in the Task 3.2. allowed the prediction of quality changes of rocket leaves and fresh-cut melon and strawberries during storage. The integration of all these results contributes to the understanding of the effects of storage conditions on overall quality of RTE vegetables and fruits and provide useful information for developing processes. Aiming at the prediction and modulation of nutritional quality and shelf-life, these kinetic parameters and physiology-based models are adequate for the development of tools as predictive softwares for the industry.
As for Task 3.3. models whose parameters can be simply obtained and that can be easily applied to different polymeric film are useful to optimize the packaging and the production process according to the packaged product features. Moreover, the respiration rate modeling is central to design modified atmosphere package intended for fresh-cut fruit and vegetables. Models of WP3 were used to optimize the packaging conditions for fresh-cut melon from Nuvifruits.
The results of Task 3.4. will provide a decision-support information for processors choosing optimal packaging solutions for RTE fruit products. For consumers, the optimized packages will provide products with extended storage life and improved sensory quality.
In Task 3.5 the survival of L. monocytogenes and E. coli after subjection to mild heat treatment and acidic conditions was studied (D3.18). The outcome of this may be of value to processors of fresh-cut foods, since it will provide information to improve processing procedures, thereby increasing safety of the product. In addition, the individual lag times of sublethally heated and acid stressed L. monocytogenes and E. coli at a range of recovery temperatures were estimated and a model was developed. This will provide a valuable tool for the food industry enable decision making and risk assessment, thus contributing to manufacture of safe products through understanding ways of controlling L. monocytogenes and E. coli (D3.19).
The software developed by DARe in Task 3.6. being easy to send to companies, is a effective way to communicate QUAFETY results to companies and increase the probability of further scaling up activities.
Potential impact for WP4
The results of Task 4.1 will be applied by growers and allow them increasing yield and improving quality of leafy vegetables with rational use of water and fertilizers. Furthermore, the new method to control the nitrate level in the vegetables will allow the growers to promote their produce as more consumer-friendly and healthy. It is important that the results have been already tested at a bigger scale at a commercial farm and the conclusions have been confirmed. For consumers, it will provide cleaner and safer foodstuffs with lower nitrate content, better nutritional quality (higher vitamin C) and slower spoilage. The latter improvement will be important also for the retailers. Thus, the development will contribute to the reduction of food losses. For policy makers, it will provide a guideline for permissible level of nitrates in soilless-grown leafy vegetables. The findings of this applied work may provide basis for a more basic research dealing with regulation of plant development and metabolism by mineral nutrition, and for breeding crops better suited to soilless cultivation.
The results of Task 4.2 are primarily important for ensuring microbiological and chemical safety of washed and ready to eat leafy greens. Prevention of foodborne disease outbreaks is equally important for the consumers and for the producers, as well as for the public health authorities. These results suggest the feasibility of substituting hypochlorite with the more effective chlorine dioxide which at low doses can reduce the risk of product contamination with pathogenic bacteria (e.g. Salmonella, E. coli H157:O7, Listeria) and with spoilage bacteria. In addition, it will prevent generation of potentially harmful and carcinogenic decontamination byproducts such as trihalomethanes in the products and in the plant wastes. The ultrafiltration technique permits to reuse the water in the process and to improve process sustainability. As an additional effect the system allows to recover some of the water used for washing, and to put it back on circulation reducing the environmental impact of the whole system being an important result for the producers and for the public and for the authorities. It is especially important that the developer of this technique is a commercial processer so that the technique has been tested at industrial-scale facility.
The results of Task 4.3 are primarily important for ensuring microbiological and chemical safety of fresh-cut melons. Prevention of foodborne disease outbreaks is equally important for the consumers and for the producers, as well as for the public health authorities. Substituting hypochlorite wash with an efficient physical thermal treatment leaving no toxic residues will reduce the risk of product contamination with pathogenic bacteria (e.g. Salmonella, E. coli H157:O7, Listeria) and with spoilage microorganisms (bacteria, yeasts and moulds). On the other hand, the treatment will make the preparation of melons for fresh-cut processing more sustainable and environmentally friendly by reducing water and energy consumption and preventing the release of toxic and carcinogenic byproducts such as trihalomehanes. The reduction of environment pollution with decontamination byproducts is important for fruit processors, authorities and public.
The results of Task 4.4 are primarily important for ensuring microbiological safety of food industry in general and fresh-cut industry in particular. Listeria is a highly dangerous potentially lethal pathogen so that prevention of its outbreaks is equally important for the consumers, for the producers and for the public health authorities. Biofilms of Listeria result in cross-contamination of the produce with this deadly pathogen. They are very hard to eradicate if not treated on time and can result in closure of a factory by health authorities. The work resulted in preparation and dissemination of recommendations to the industry for practical measures on preventing the buildup of Listeria biofilms on the surfaces of fresh-cut facility. These recommendations are important also for policy makers for ensuring public health.
The results of Task 4.5 will be of primary importance for fruit processors due to the increasing efficacy of cutting and peeling operations. It will improve work conditions and the safety of the technological process by preventing manual operations with sharp knives under worker-unfriendly conditions (low temperature, decontamination chemicals). Exclusion of a human factor as a potential source of contamination will improve the product microbiological safety. On the other hand, substitution of manual operations with automatic ones will allow running them under strict conditions e.g. under UV light and low temperature further reducing the chances of product microbial contamination with spoilage or pathogenic microorganisms. The latter factor is important for processors, consumers and health authorities. The work has resulted in submitting a provisional patent application that has already attracted the interest of machine-building industry that is another important stakeholder in the fresh-cut technology.
The results of Task 4.6 will improve quality and reduce spoilage of ready-to-eat products, in particular shredded Chinese cabbage. These results are important for vegetable processors, retailers and consumers. Controlling the product spoilage will contribute to the global task of reducing food losses. For researchers the findings of this applied work may provide basis for a more basic research dealing with regulation of senescence and metabolism in plant tissues by physical factors such as temperature.
The results of Task 4.7 are primarily important for ensuring microbiological and chemical safety of ready-to-eat leafy greens and fresh-cut fruits e.g. melons. Prevention of foodborne disease outbreaks is equally important for the consumers and for the producers, as well as for the public health authorities. Strengthening antimicrobial measures by additional hurdle of physical intervention (UV light) immediately before packaging will reduce the risk of product contamination with pathogenic bacteria (e.g. Salmonella, E. coli H157:O7, Listeria) and with spoilage microorganisms (bacteria, yeasts and moulds). The latter factor may be important for both retailers and consumers and contribute to the global task of reducing food losses. For researchers, the findings of this applied work may provide basis for a more basic research dealing with regulation of metabolism in plant tissues and modulation of host-pathogen interactions by physical factors such as UV light.
The results of Task 4.8 are important for ensuring microbiological safety and preservation of quality of ready-to-eat fruit and vegetable products (leafy greens and fruit) and therefore are of special importance for the processors. Prevention of foodborne disease outbreaks is important for the consumers and for the producers, as well as for the public health authorities. The development of active antimicrobial packaging and coating will reduce the risk of product contamination with pathogenic and with spoilage microorganisms (bacteria, yeasts and moulds). The latter factor may be important for both retailers and consumers and contribute to the global task of reducing food losses. The novel Time-Temperature indicator may be of interest for retailers, consumers and decision makers providing a more accurate instrument to evaluate the goodness of the product on the shelves. It will prevent both unnecessary rejections and wastage, on one hand, and keeping the unmarketable products on the shelves, on the other.
The results of Task 4.9 will allow the maintenance of cool conditions throughout the supply chain, from the field and to the final consumer and therefore will be of interest for all stakeholders: growers, processors, distributors, retailers and the consumers. The containers developed in this task will improve quality preservation and reduce proliferation of microorganisms on leafy greens eventually controlling their spoilage. This will contribute to the global task of reducing food losses. Controlling microbial proliferation will prevent the potential pathogens of reaching an infective dose and this will improve the produce safety reducing the risk of disease outbreaks. It is especially important that the developer of the container is a commercial processer so that the appliance has been tested under industrial conditions.
Potential impact for WP5
WP5 represents the most relevant WP in regard to the impact of main results of QUAFETY project. In fact in this WP a number among the most promising results from previous WPs were tried at a larger scale and often their application was demonstrated at industrial sites, including processors internal to the Consortium, and a number of external entities, including private producers and processors, and public health authorities. Apart from being a necessary step toward potential implementation of the innovations into the “real world” the other important thing is that participants in this Project were the first to whom results were disseminated, and in the most effective way. In this way results dissemination/exploitation started within the Consortium and this in turn have enriched the same results with feedbacks deriving from this first phase of “internal” dissemination. In addition it is worth noting that in sectors that are disjointed as the agricultural and the food system, the introduction of a systemic approach for food quality and safety is of strict relevance. Furthermore, WP5 highlights the importance of relations between the different components of the system. The cohesion between the different organizational objectives is achieved only by acting with a system able to ensure the integration of the activities carried out by different actors.
Potential impact and main dissemination of WP6
WP6 focused on testing the hypotheses of ready to eat fruits and vegetables as substitutes for conventional whole products, and quality attributes deriving from innovative production technologies, within those produced by QUAFETY research, in order to understand consumer preferences and draw consideration on the consequent consumer acceptability and measuring consumer perception towards these attributes. In addition, consumer acceptability measures were used for setting up the cost-benefit analysis of the innovations produced by QUAFETY. Thanks to these activities DARe staff were in touch with a series of companies which showed deep interest into the economic evaluation of the innovations. They most common statement has been “usually we implement innovations and make things work out, with such approach we have a prevision of what would be the impact on our cost structure and price potentiality”. The main impact of this activity is related to the possibility to use this approach to enhance communication between scientists and stakeholder, and to further develop this tool to drive innovation processes.
Potential impact and main dissemination WP7
The WP7 results are addressed to four basic groups of users:
1. Stakeholders from the food sector as end-users of final application including: food industry, consultancy services, retail business
2. Public food safety authorities
3. Researchers in the field of food science, agribusiness and horticulture
4. ICT companies for the development of new applications for agriculture and food sector, mainly using implemented tools, encompassing: SME developers of ICT for agriculture platform and technology producers.
The food sector is of strategic importance for European society and economy. Due to its complexity, food operators have to manage many different and heterogeneous sources of information. The tool will help fresh cut food sector to collect, storage, share and analyse data related to quality and food safety. The potential impact of application the specific tools developed in WP7 that is, the questionnaire and the safety assessment plan (SAP), as an integral part of the quality management and food safety management system in a SME is of great importance. These could be used as a self-assessment tool in the SMEs and as an internal audit tool, providing specific quality tools to the fresh cut produce sector for their reviewing managerial processes. Results obtained by the SAP will determine the actual hygiene and safety level of the SMEs and will validate the best practice score that will be assigned to SME after interviewing. The socio-economic impact is that SMEs will improve their quality and food safety level via intervention in conjunction with the procedures developed in the project and incorporated in the FSMS guide. The societal implication is evident by offering fresh produce of a better quality and safety to the final consumer.
Main dissemination numbers
Dissemination was one of the strong points of Project. QUAFETY activities and results have been presented at various level in a very hign number of Countries, both in Europe (Fig. 28) and outside (Fig. 29). With reference to the previously described results in WP8, following a synthesis of main dissemination numbers are reported according to the type of stakeholder target (R&D, industry and authorities, media and consumers).
Dissemination activity targeted for R&D. Up to date a total of 24 peer-reviewed scientific papers have been published within journals, edited books, and conference proceedings; of these, 15 proceeded from activities in WP1, 4 from activities in WP2, 1 from those of WP3, and 4 from those of WP4. A prudential estimation considers at least 20 more papers being published within the next 12-18 months mainly from WPs 2, 3, and 4. In addition a total of 13 dissertation Thesis have been produced within the Project, mainly at AUA and UNIFG.
Participation of QUAFETY scientist in international scientific conferences allowed the presentation of 48 oral contributions (11 of which as invited keynote) and a total of 44 posters.
Finally, “special QUAFETY sessions” have been organized within 4 major scientific conferences in Italy, Portugal, and Poland.
Dissemination activity targeted for industry and authorities. A very large effort was made in order to diffuse the outcomes from the Project to the produce industry, with particular emphasis to fresh-cut produce companies. With this objective a number of strategies were used. More than 500 articles published in the technical and popular press and websites, mainly in English, Spanish, and Italian. In addition results were illustrated in about 55 oral presentations in industry meetings.
QUAFETY scientists participated as instructors in industry targeted educational events in Turkey and in UK while 4 specific workshops were organized for a public audience in Italy, Greece and the Netherlands, and within the popular international exhibition Fruit Logistica in Germany.
In some of these events informative material was freely distributed to participants in form of flyers, USB pendrives, and CD-ROM.
Additional dissemination activities were performed through the world wide web with the project website (Fig. 30) and the fresh-produce industry interactive platform Tacler (www.tacler.com).
Dissemination activity targeted for media and general public. Besides the QUAFETY website, a Youtube channel was available to download video documents produced within the Project. In addition UNIFG Facebook page hosted all kind of news related to the Project for about 1,300 contacts.
Finally a number of interviews were released by QUAFETY scientists to different media type, both web-based and traditional, at local, national, and international level.
List of Websites:
Project website
www.quafety.eu
Project Coordinator (Prof. Giancarlo Colelli)
giancarlo.colelli@unifg.it
