Final Report Summary - FISHFERMPLUS (Added-value bioprocessing for Fish raw materials via low-cost Fermentation technologies delivering a PLUS in the sustainable production, consumer safety and quality of highly sensitive fish products)
Executive Summary:
In fish filet production almost one third of the high-value catch is lost, often sold as cheap animal feed. Aim of the FishFermPlus project has been to develop a process enabling more sustainable handling of scarce aquatic resources and a more economically efficient use for fish manufacturers. This is also of ecological and ethical interest in times of overfishing and of high protein demand of a growing world population.
Therefore, a bioprocess has been developed which allows the utilization of high-quality by-products from filet production. The process results in products with improved sensory characteristics on the one hand and increased safety and shelf-life on the other hand.
In order to achieve the envisaged objectives, the project consortium conducted an extensive literature review and identified by additional stakeholder consultations relevant market needs. The conclusion was that shelf-life prolongation is the most important goal for fish processors to increase quality and added value of their products.
In the following, suitable raw materials and cultures were tested to establish an effective bioprocess. In addition, optimal fermentation parameters were identified, which guarantee the production of value-added products produced in an economically efficient manner.
The fermented products were further processed, in order to obtain stable, marketable products. The stabilization steps included texturing processes, addition of antioxidants, salting as well as drying and heating treatments.
The stabilized intermediate products were finally incorporated in product applications. These comprised convenience products like spreads, burgers or filled puff pastry. However, it was shown that the products can also be used as ingredients e.g. as pizza toppings.
Safety assessment of the flexibly applicable intermediate product has shown that during the final heat treatment non-spore forming pathogens are efficiently killed, while spore-forming bacteria cannot grow during storage. Thus, with regard to food safety, the fermented and stabilized product was stable at room temperature (22.5 °C) during the observed storage time of 32 days.During chilled storage (5 °C) no pathogenic growth could be observed during 42 days. According to predictions including variability in product characteristics, contamination with Listeriaafter heating could lead to growth from 1 CFU/g to 100 CFU/g (critical limit) at 5 °C in 190 days with a lag time of 77 days. Considering the possibility of no lag time of Listeria and even larger variability in product characteristics, a safe shelf-life of 2 months was recommended.
Models have been developed which can be used to predict pH, lactic acid concentration as well as pathogenic growth during fermentation based on fermentation parameters. Models for heat inactivation are helpful to calculate effectiveness of different heating parameters with regard to reduction of pathogens. Further models are available to predict growth of Listeria during storage.
Sensory and chemical tests have shown that no off-flavours from fat oxidation occurred and that furthermore peroxide value (PV) and thiobarbituric acid reactive substance (TBARS) values did not increase during storage of the fermented and vacuum packed salmon product, suggesting that chemical changes during storage up to more than one month may not be the limiting factor for distribution of the product.
Pilot trials were performed in the facilities of the SME partners. Trials have indicated that upscaling of the process is realizable in an industrial setting. Furthermore, reproducible results compared to laboratory results were obtained. Thus, it is expected that further upscaling to industrial scale will be possible without remarkable challenges. Shelf-life tests of developed spreads have proven good product quality for at least two weeks.
During demonstration events the developed products were presented to potential customers. Furthermore, the opinion of the broad public was collected during a consumer test. A very positive feedback was received at these tastings which gives a promising indication for the products’ market potential.
During the post project phase further optimization with regard to final upscaling and industrialization of the developed process will be performed by the SME partners. Expected time to market of the products is one year after project end.
Project Context and Objectives:
The European fishing industry is one of the largest in the world. However, it faces strong competition from Asia and South America, with India and Russia catching up quickly. Despite a production of more than 5 Million tonnes of fish each year, the internal fish market tends to be not well-balanced: exports were made competition- and price-driven at a lower average price unit than imports for high-price raw materials.
This context shows clearly the need to create value in the internal market by serving for top quality demands, and by a clear distinction from cheap production markets. Such an approach requires real added-value with a tangible plus in terms of fish and seafood quality, safety and convenience. A mere price-/production unit-driven approach will not succeed for the European fish industry as the Union cannot compete with the production areas, high input aquaculture systems and coastlines of e.g. China, India and Russia, and has itself also committed to a more sustainable fish production with restrictions in catch and fleet quotas. Innovative alternative approaches are demanded since more than 350,000 jobs in the European fish industry are at risk, which are typically found in small- and medium-sized companies (SMEs) and account for almost 9% of the total European food industry, a major and top-priority pillar of European and rural economies.
Hence, addressing market mechanisms and consumer interests successfully will become a must to cope with future developments: Consumers were bombarded with news about foodrelated health crises around Europe which left them feeling insecure about the quality and safety of the foods they purchase. Changing lifestyles, work patterns, rising disposal income have resulted in more nutritional and health conscious consumers demanding top quality foods at utmost convenience for their use and storage. But also price remains an important driver in many European markets; and longer shelf- life is a key factor for high convenience and flexible supplies and has thus become indispensable.
Fish and seafood products can cater for many of these trends, but also face severe challenges from the latest developments: On the one hand, fish accounts for a “healthy food” fitting well into a balanced diet and delivering omega-3-fatty acids and other nutrients, which are considered to fight degenerative, age- or lifestyle-related diseases (Alzheimer’s, CVD etc.). On the other hand, it is a highly sensitive and perishable food, which requires accurate cold chain management and hygiene measures against food-borne diseases. It may even evoke fears of contaminants or other risk factors, such as sea pollutants, antibiotics etc. Analytical regimes and hygiene management systems are well developed and a number of technical singular solutions (e.g. packaging, preservatives, irradiation, canning, curing etc.) exist, but not all of them are evident to the consumer or well-appreciated as they do not match their demand for high-quality minimal processed foods, natural processing and ingredients, or waste minimisation. Instead guidance from simple symbols and reliable quality promises are sought-after as it can be observed from the MSC seal indicating sustainable production systems, or the Fresenius tick indicating compositional quality.
Next to the key challenge of the difficult product and market positioning to reach consumers, each fish processing company also faces very tangible day-to-day challenges arising from the operational and economic view-point of the business, which are again linked to a number of societal objectives and sustainable development as mainly the resource-efficiency in the fish processing is concerned. In the light of recent reports which point at the earlier (than previously estimated) arrival of the "fish dependence day", where Europe has nominally consumed all its own fish and needs to bring in stocks from elsewhere, it seems absurd that currently 30% of the initial catching are discarded and lost for human consumption along the total fish supply chain. Main factors accounting for this loss are frequently marketing requirements (portion sizes, trimmings, no visible mechanical damage of the fish flesh) towards a specific, intended use, and not food quality or safety concerns. The loss of resources and food wastage even continues beyond this figure in retail and consumer homes. Due to the short shelf-life and fear of food-borne diseases, restrictively set best-before-dates, and no checks of the actual food quality combined with inadequate stocking, many products are simply thrown away. The latter context may be basically out of the direct control of a fish producer, but affects his economic viability, if the shelf-life of his products is found to be too little to suit the retail and consumer needs.
Gaining a competitive edge in this context therefore means (a) to contribute against food wastage and towards sustainable development, (b) to fully utilise own resources optimally and (c) to provide real added-value products, which are distinct on the market in terms of food safety, quality and convenience/shelf-life. The latter cannot be achieved via chemical additives or extreme processing as done so and targeted in the past, because this will not achieve the crucial consumer acceptance and buying result on the market. The utilisation of available raw materials as suggested in (a) and (b) requires perspectives in addition to low-cost valorisation (e.g. minced sales, pressing).
The SME beneficiaries of the FishFermPlus project strongly believe in cost-efficient, simple, physical processing, such as fermentation, to deliver solutions against these needs. They see the potential in this approach to be inspired by cutting-edge biotechnological knowledge, but to deliver down-to-earth implementation solutions, which are broadly usable and clearly market driven. Hence, a balanced consortium of technology-driven SMEs supporting the food and fishindustry and of SME fish producers have gathered to outsource research activities to developand design such a process, but to remain with the full development power over the concertedprogram of activities in a Research for SMEs project.
The overall objective of the FishFermPlus project was the development of a bioprocess which generates new properties by treatment of fish raw material and by-products of fish filet production. In this way, an added value has been created, by improved sensory characteristics as well as by enhanced safety and shelf-life. In the end, additional economic value will be created for fish-processing companies by valorisation of underutilized material and food wastage can be reduced.
In order to succeed with this overall mission, the FishFermPlus project needed to fulfil a number of technical and operational objectives:
(1) A clear definition and specifications of the actual new process design had to be achieved by a bottom-up approach for the compilation of detailed requirements. These had to be evaluated positively by the end-users included in the FishFermPlus consortium and also in the frame of confidential in-depth consultations with representatives from the fish (processing) industry. Despite having gained insight into technological needs, opportunities, cost restraints and actual real business capacities by various methods, such as market reviews, online surveys, literature reviews and in persona consultations, a conclusive blueprint for the development was elaborated.
(2) The identification of suitable strains, cultures, substrates and other ingredient components to promote desirable microbial growth and the production of target metabolites was most crucial for the whole project success. For this purpose a highly efficient screening method was implemented to maintain development speed and result-orientation in the identification process, the selection and exploration of suitable combinations, as well as the latter optimisation.
(3) Once a viable and efficient microbiological system had been developed for the fermentation of fish substrates, finding an optimal balance for the stabilisation of the FishFermPlus ingredients was required in order to arrive at industrially applicable products. Under this objective, the need for sensitive and gentle product processing (to maintain functionality, sensory properties and overall shelf-life of the ingredients) had to be addressed as well as the final costs and operability of the process.
(4) The next crucial operational objective revolved around the detailed exploration of the targeted ingredients/compounds/intermediate products and an in-depth investigation of their effect on the manufacturing of new fish and seafood products. It was envisaged to arrive at a minimum number of 3 added-value application processes, which will have significant economic impact for the end-user SMEs. To this end, it was foreseen to achieve and to document the development and validation of re-texturized products to be used as spreads, fillings, toppings or bratwurst-like sausages. The performance of the ingredients/compounds/intermediate products and the actual characteristics in terms of antimicrobial, flavour-/texture-improving activity or prolongation of shelf-life needed to be verified and validated at various development stages.
(5) As market-oriented project, FishFermPlus further aimed at contributing to underpinning quality assurance measures by evaluation of models for product changes based on data obtained in pilot validations. Such models will be an important building block towards the future marketing of the new FishFermPlus products and could potentially be used in terms of technology transfer to new customer companies. A specific task has been dedicated for assessing all data and performing safety assessments by applying relevant modelling approaches.
(6) Physically, FishFermPlus aims at delivering a bioprocessing system to the market. Hence, crucial technological and operative objectives concern also the integration of potential batch sizes, desirable production times, scheduling and logistics, energy costs etc. in order to enable the latter broad implementation and exploitation. It is expected that a latter investment in the bioprocess by future customer companies should not exceed € 30.000 in order to promote the up-take. Next to the measurable cost factor, the functionality of the system will also be fully assessed by testing and validation of the FishFermPlus technology in real environments with products of market concern and value, which will deliver documentation for the FishFermPlus added-value beyond the scope of scientific small-scale trials. The scope of the project enfolded technological requirements and benefits, as well as considering socioeconomic factors.
(7) The final operational objective relates to the necessary pre-requisites for realising the targeted impact and return-on-investment by a broad support of the up-take of the FishFermPlus results by SMEs and a wider audience. The successful accomplishment was and will be supported by carrying out a comprehensive series of demonstration and training activities which will prove the viability and outline the economic potential due to real product quality advantages.
Project Results:
WP 1 - Stakeholder Targeting and Definition of Industrial Specifications
Task 1.1 European-wide market review and in-depth consultations
A short questionnaire was developed by DTU in order to identify a representative opinion of the fish sector regarding the FishFermPlus objectives of shelf-life prolongation and sensory improvement of fish products. The questionnaire was available online and was translated by DTU into French and by TTZ into Spanish, in order to improve the response of the Mediterranean fish sector, which is not represented in the project consortium. In addition to addressing concrete companies, also associations were contacted by DTU and TTZ as multipliers which were asked to forward the questionnaires to their members. In total, 170 Spanish, 45 French as well as 393 further European stakeholders were contacted and asked to answer the questionnaire. The response rate was quite poor, only 15 questionnaires were filled. As experienced by the beneficiaries in other projects, it is generally hard to convince companies to take part in such a survey, due to the fact that they are too busy with their daily business and are afraid of revealing confidential business information.
The analysis of the collected responses results in the following prioritized list of needs and expectations:
1) Antimicrobial properties of a fermented ingredient are important (High importance: 86 % of responses) to obtain a shelf-life extension of 10-50%.
2) Texture improving properties of a fermented ingredient are important (High importance: 57 %) to reduce liquid loss and to facilitate slicing (and reduce slicing losses).
3) Flavour enhancing properties of a fermented ingredient are important (High importance: 43 %) primarily to reduce off-odour and off-flavour development.
4) A fermented ingredient is interesting even if it has to be declared e.g. as ‘protective culture’ (71.4%).
The exact results of the market review can be found in D1.1.
In addition to addressing the European industry, extended questionnaires were sent to the SME partners ISI, QFP, RAUCH, and BG. These were accompanied by personal consultations from DTU and TTZ in order to obtain more information about the SMEs’ interests regarding specific raw material and properties they would be interested in. This information was also used for identification of processing and application specifications in tasks 1.4 and 1.5.
In addition to the results from the market review, the interviews with the project partners revealed that for low-price products, like those of BG, shelf-life extension is of most importance. Whereas for high-price products and especially for products targeted to the catering and gastronomy sector, which do not need a long shelf-life, the sensory properties are more relevant.
Task 1.2 Literature research
An intense literature review of latest scientific findings in relation to fish fermentation was performed by DTU. As discussed with the whole consortium at the kick-off meeting, this was mainly focused on the traditional Asian market, since it is the largest market for fermented fish products worldwide. The report contains information about types of fermented fish, microorganisms detected in fermented fish as well as their properties, texture and flavour compounds in fermented fish, shelf-life and food safety issues of fermented fish, biopreservation methods as well as commercial cultures. However, hardly any studies could be found with regard to controlled, standardized fermentation, which underlines the high innovative approach of the FishFermPlus project.
The whole literature review can be found in D1.2.
Task 1.3 National and international standards and legislation requirements
TTZ supported by DTU compiled an overview of legislations and standards of relevance for the FishFermPlus project. Support came from the partners RAUCH and BG who reported about requirements in their productions. The report contains aspects about conditions for using living cultures in food, microbiological criteria in food, especially concerning listeria and histamine, bacteriocins in food, labelling requirements, novel food regulations as well as the Good Manufacturing Practices.
This comprises EFSA’s QPS-list of cultures considered as safe, the inventory of microorganisms with a documented history of use in food published by the International Dairy Federation and the European Food and Feed Cultures Association, Commission Regulation (EC) No 1441/2007 on microbiological criteria for foodstuffs containing limits for L. monocytogenes and histamines in fish, labelling requirements according to Directive 2000/13/EC as well as current Good Manufacturing Practices (GMP) published by the US Food and Drug Administration with special focus on microbial food enzyme production.
The results are presented together with the literature review in D1.2.
As demanded in the DoW, fish sector associations were contacted by DTU and TTZ in the frame of task 1.2. However, for the legislation report, the aspects to consider were focussed on the work to be performed and developments planned during the project rather than on the general standards implemented in industry.
Task 1.4 FishFermPlus processing requirements & specifications and Task 1.5 FishFermPlus application and end-user requirements & specifications
As described in D1.3 SME partners found it difficult to define exact processing and application specifications, since especially the pricing depends on the real outcome and benefits achieved during the practical trials. The final type of the fermented products is not clear yet either, as final stabilization methods still need to be tested. Thus, concrete threshold characteristics like pH and aw-values are hard to define at the current stage of the project.
However, the SME partners ISI, QFP, RAUCH, and BG supported by the RTDs TTZ and DTU worked out possible strategies that could lead to products of interest. The concept comprises two approaches. One attempt is tailored for high-quality raw material like filets as well as bits and pieces and aims primarily at the ripening and creation of new sensory characteristics. On the other hand, low-value fish mince will be fermented to achieve mainly shelf-life and safety improving properties. Both approaches are described in D1.3 and are the basis for practical trials in WP2.
WP2 - Development of the fermentation process
Task 2.1 Experimental investigation design and overall development approach
For development of the fermentation design numerous trials were conducted (D2.2). For the fermentation repository it proved successful to ferment in plastic bags with controlled vacuum. For blue marlin a vacuum of 70 % was identified as optimal parameter whereas salmon mince was fermented at 25 % vacuum.
Selection of fermentation substrates, starter cultures as well as fermentation parameters is described below.
Control parameters to compare successful fermentation and reproducibility of the process were pH of the raw material, inoculation cell count as well as pH, LAB cell count, and smell after fermentation.
Task 2.2 Substrate selection
Minced salmon was fixed as substrate for fermentation since it is the main by-product from filet production at BG’s production process.
Since tuna proved unsuitable for fermentation in first trials, further fish species of high relevance in the business of RAUCH were selected, namely wild Alaska salmon and blue marlin. Finally, blue marlin was selected due to better sensory results during fermentation.
Task 2.3 Strain identification and culture development
Selection of strains to be tested was done jointly by TTZ, DTU, QFP, and ISI according to results from the literature review in WP1 as well as based on experience by ISI and QFP. Only food-grade cultures were selected. In total, 26bacteria for acidification as well as seven yeasts for flavour development were tested.
Strains were provided by ISI or bought from the German Collection of Microorganisms and Cell Cultures (DSMZ) and Chr. Hansen. All cultures were included in TTZ’s strain collection and were cultivated by TTZ.
In addition to single strains also fermented food products were used as fermentates in order to test their influence on flavour production. The following products were used:
- Sauerkraut juice
- Bread drink
- Kombucha
- Water kefir
- Natural yoghurt
Identification of final starter cultures is described under task 2.5.
Task 2.4 Final selection of processes and raw materials
For the blue marlin bits and pieces, L. diacetylactis was identified as most effective flavour producing strain. The developed butter flavour was clearly recognizable and evaluated by RAUCH as interesting new product characteristic that will add value to the bits and pieces and create a unique selling proposition of later products. Combination with 0.25 % ascorbic acid was most effective in intensifying the butter flavour. Further parameters defined include 2 % glucose, 0.005 % booster, 20 h fermentation time, fermentation in evacuated bags with a vacuum of 70 % and a fermentation temperature of 25 °C.
For the salmon mince, L. diacetylactis could produce a pleasant flavour as well. Flavour characteristics could be improved by 10 minutes cooking prior fermentation and by addition of 0.25 % citric acid. Addition of 2 % glucose, 0.005 % booster, 20 h fermentation time, fermentation in evacuated bags with a vacuum of 25 % and a fermentation temperature of 25 °C proved effective also in this case.
Task 2.5 Process optimisation and definition of parameters for transfer
During optimization trials different strains of L. diacetylactis were investigated. Thereby, different results were achieved for blue marlin and salmon mince with regard to sensory characteristics and intensity of the produced butter flavour. Therefore, two individual starter cultures were selected, DSM 20384 for blue marlin as well as DSM 20729 for salmon mince. During stabilization trials in WP3 these were produced as freeze-dried cultures for easier handling during upscaling trials and use in an industrial environment in WP3 and WP5.
Furthermore, for reproducibility of pleasant sensory results it was decided to ferment with L. diacetylactis alone and not in combination with other strains as indicated in D2.2.
During reproducibility and validation trials the impact of raw material quality became evident. Problems with blue marlin quality on the world market had a negative influence of sensory results after fermentation. Therefore, the optimization development became delayed with regard to original project schedule. For later commercialization it is highly recommended to ensure raw material quality for reproducible fermentation results. Further aspects that led to a delay in fermentation optimization due to insufficient reproducibility results included the need for better standardization of processing steps and their adaptation by both RTD partners.
For salmon mince it was obvious that the material was highly perishable due to its large surface and the high oil content with regard both to microbial spoilage and fat oxidation. Optimization and standardization of the raw material quality could be achieved by direct evacuation and freezing after production.
Calculated inoculation concentration was slightly increased from 106 cfu/g to 3x106 cfu/g, since trials had shown that actual cell count in the product was always a bit lower.
WP3 - Downstream Processing & Equipment Design
Task 3.1 Selection and optimization of stabilization technologies
The original objective was to identify stabilization processes for the fermented products in order to obtain stable and functional fermentates for later compound development and use of these in different product applications.
However, drying tests in WP3 have shown that due to the high oil content of the salmon mince from BG it was very challenging to obtain a powdery product. Even with high percentage (up to 80 %) of the carrier substance maltodextrin the result was not satisfying. Salmon mince is the major material occurring in high amounts at BG and there is no other material of interest for them to replace the salmon mince. Therefore, the partners agreed not to continue the approach of drying and compound development, since the development of further procedures would be too time- and resource-consuming. Furthermore, the resulting process was likely to be too sophisticated and expensive to be feasible and efficient for later industrial implementation. Also the oil removal was not favoured, since it would mean an additional processing step. At the same time, the aim of utilizing the material completely would not be reached.
Therefore, the original goal of drying for further use as fermentates had to be modified as part of the DoW amendment n°1. While the developed starter cultures are available to be offered as fermentates, main new task was the stabilization of the fermented products for further development to intermediate products and related product applications. Thereby, the restructuring of the bits and pieces as well as of the salmon mince became the main focus together with avoidance of fat oxidation.
Since trials revealed that fermented blue marlin pieces were too instable to glue them with economically efficient concentrations of transglutaminase. Therefore, it was decided to ferment whole filets which can be sold as high quality products, which even increased value on the one side. On the other side, bits and pieces should be fermented as cubes which can be sold e.g. as salad toppings.
Drying trials were started with whole filets. Tests were performed directly in RAUCH’s facilities in order to get realistic results with their equipment and conditions. Furthermore, this was more efficient with regard to later process integration and scale-up. Identification of optimal parameters was mostly based on RAUCH’s experience in other product applications. Trials revealed that drying of products without addition of smoke was more promising, since sensory evaluation indicated that the subtle fermentation flavour was completely covered by the smoke aroma. The selected processing parameters comprised a ripening step with salt and additional spices for 48 h at 2-4 °C followed by a drying step for 7.5 h at 26 °C at a humidity of 30 %.
For salmon mince a process was established that transferred the inhomogeneous mass into a smooth, spreadable product. Therefore, an emulsification process was developed that included addition of oil, water as well as a mixture of stabilizers.
Since BG expressed also interest in a sliceable texture, further trials were done with modified ingredient concentrations. However, the result showed that due to the denaturation of proteins during the cooking process prior fermentation, the bonding of proteins was hampered. For maximum benefit of BG and later utilization of their salmon mince, further trials were conducted with unfermented mince. With the uncooked material a sliceable bologna-like texture could be achieved with the developed stabilizing agents. This will offer advantages for different applications, although without the special benefits targeted at by fermentation like improved safety and shelf-life.
Another challenge of the salmon mince resulting from the high fat content was the product’s susceptibility to fat oxidation and related unpleasant sensory rancidity notes. QFP has developed a compound made of grinded rosemary which is known for its antioxidant capacity.
Task 3.2 Process integration and scale-up
In preparation for the pilot validation, first up-scaling tests for the salmon mince fermentation developed in WP2 were conducted at TTZ’s facilities. Heating in a heating reactor with stirrer was performed with 7 kg of salmon mince raw material. For mixing of fermentation ingredients a cutter was used with the same capacity. Incubation was done in 1 kg bags.
Based on developments from task 3.1 and WP4, upscaling trials for salmon mince stabilization and product application were performed as well. A batch size of 5 kg was processed in a HydroGrind allowing high shear forces with rotations of 90 Hz. The resulting products showed a more homogenous emulsion compared to lab trials by QFP as well as stable properties after freezing for several weeks and subsequent defrosting.
Results confirmed those achieved at lab scale with regard to growth of starter culture and pH decrease. Thus, first scale-up trials indicated an easy upscalable process.
Products produced during the upscaling trials were used for the first external demonstration event at the project’s Seafood Expo stand (see WP6) as well as for safety assessment trials by DTU in WP4 and 5.
During the planning of the pilot trials, safety assessment by DTU revealed that the fermented blue marlin is not sufficiently safe. The development of an additional process for stabilization of blue marlin was not feasible within the remaining time. Therefore, it was decided not to perform pilot validation of blue marlin. Instead, RAUCH expressed the interest to test salmon mince processing in their plant as well.
More details are described in D3.1.
Task 3.3 Pilot validation design and building
After successful first upscaling tests, next step was the transfer of the whole process to pilot scale in the plants of the producer SMEs RAUCH and BG. As described above, it was decided not to perform pilot validation with blue marlin, as initially planned. Therefore, pilot validation of the salmon process was planned both for RAUCH’s and BG’s facilities. Pilot validation design was the basis for trials to be performed in WP5.
Due to extensive discussions between the SMEs and TTZ it was guaranteed that all chosen equipment was appropriate for a promising conduction of pilot trials and reproducibility of lab-scale results. Due to sufficient actual testing environment, it was not necessary to assemble any additional equipment. Additionally, no specific adaptions of available equipment were needed.
Aim of the pilot trials was to validate that the results achieved in realistic industrial surrounding are in accordance with laboratory results. Therefore, an analysis protocol was developed for recording pH values, cell count, heating profiles as well as smell.
More information about the selected equipment is given in D3.2.
WP4 - Development of Product Applications
Task 4.1 FishFermPlus intermediate products development
Stabilized salmon mince developed in WP3 were used as basis for further development of the intermediate product. As a result, a fish flavour ingredient has been developed containing a smoke substance for support of antimicrobial activity and spices that harmonize fish the aroma from the fermented salmon and the rosemary flavour from the Antiox applied during stabilization. A process was established for combined application of stabilization and flavour ingredients. The resulting product can be used either directly as a spread or as an ingredient in other product applications. All market needs identified in WP1 including texture improving properties, flavour enhancing properties, and a fermentate for application in fish processing could be met. The last aspect of shelf-life extension is described below. In addition, price calculation revealed that the developed product can be marketed at a profitable and competitive price.
Task 4.2 Product application development and validation
The intermediate product developed in task 4.1 was further developed for several product applications. Since RAUCH is selling in the premium catering sector focussing mainly on unique products of high quality, while BG distributes products in a lower price sector, different product application were developed with both partners.
For BG main interest is to sell the intermediate product as ingredient for convenience products like salads, pizza, puff pastry etc. For using it as slices on pizza or cubes on salads a sliceable texture was desired by BG. As described in D3.1 this could not be achieved completely. However, trials performed by QFP in cooperation with TTZ and BG revealed that the texture was solid enough for the mentioned applications. In addition, several convenience applications were tested and proven feasible like fish hamburgers, fish nuggets, and salmon filled pastry. The intermediate product could be shown to be stable in texture during baking and frying.
Due to the challenges that occurred during texturing of blue marlin pieces (see D3.1) it was decided to ferment whole blue marlin filets, in order to allow applications for RAUCH’s desired raw material and to be able to present products at the already planned Seafood Expo. RAUCH developed three types of seasoning, namely natural, sesame, and pastrami, which matched and did not mask the fermentation flavour.
Due to the results from safety assessment, that revealed that fermented blue marlin is not safe enough, RAUCH decided to develop product applications for the salmon mince as well. Since RAUCH delivers products mainly to the catering and gastronomy sector, they developed a convenience product e.g. for bakery to-go products. In total, they developed four spreads containing next to the FishFermPlus product, smoked salmon cream, mayonnaise and partly roe. These products were used for pilot production and consumer evaluation in WP5.
Safety assessment of intermediate salmon product
Microbial safety of the fermented salmon mince product was assessed. The fermentation process was well characterized including growth of added starter culture (lactic acid bacteria), change of pH and formation of lactic acid during fermentation.
Challenge tests were performed for three key steps including fermentation, heating to improve product texture and final chilled storage. Growth and survival of seven pathogenic bacteria, including Bacillus cereus, non-proteolytic Clostridium botulinum, E. coli, Listeria monocytogenes, Salmonella, Staphylococcus aureus and Yersinia enterocolitica were assessed. Concentrations of pathogenic bacteria increased 2 to 4 log CFU/g during fermentation at 25°C except for non-proteolytic Cl. botulinum where 1 to 2 log-reductions was observed. Starter lactic acid bacteria reduced but did not prevented growth of pathogenic bacteria during fermentation at 25°C. Heating of fermented salmon mince reduced concentrations of the studied non-spore-forming pathogenic bacteria by more than 6 log CFU/g and of spore-formers by 1 to 2 log CFU/g. This heating step was important for safety of the product (3.3 in the Appendix). None of the evaluated pathogenic bacteria were able to grow in the fermented product at 5 °C but at 22.5 °C, representing room temperature, growth was exclusively observed for L. monocytogenes, Salmonella and St. aureus. The fermented and textured salmon mince product was found to be sufficiently preserved to allow distribution under chilled and room temperature conditions when the product was heat treated in the final and sealed packaging used for distribution. If the product was processed including repacking after the heat treatment then contamination and subsequent growth of L. monocytogenes, St. aureus and Salmonella were identified as risk with respect to safety of the fermented salmon mince product. Observed growth or survival data were compared with predictions from available models and in general these models had poor performance. New models were developed for growth of lactic acid bacteria and related lactic acid formation and pH reduction during fermentation as well as for growth of L. monocytogenes during storage of the final product. These models were used to evaluate the effect of variability in product characteristics on safe shelf-life of the fermented salmon mince product. By using L. monocytogenes as safety indicator bacteria, and evaluation of variability in product characteristics of 6 samples from different batches, the safe shelf-life of the salmon intermediate product could be determined as 76 days at 5 °C from assistance of the model.
Safety assessment of blue marlin during fermentation
Challenge tests for microbiological safety of fermented blue marlin meat were performed. Growth and survival of three pathogenic bacteria, including L. monocytogenes, Salmonella and St. aureus were assessed. The methods used for challenge testing of fermented blue marlin meat were similar to those used for fermented salmon mince.
The fermentation process at 25°C resulted in unacceptable growth of all three investigated pathogenic bacteria of 2.5 to 4.0 log CFU/g. Starter lactic acid bacteria (LAB) reduced but did not prevented growth of the pathogenic bacteria during fermentation at 25°C. Growth and interaction for starter LAB and both Listeria monocytogenes and Staphylococcus aureus seemed to correspond to the Jameson effect. The observed growth/survival of starter LAB and the three pathogenic bacteria was compared with predictions by models developed for the salmon intermediate product. Models for the salmon intermediate product provided relatively precise predictions for growth of L. monocytogenes and St. aureus during fermentation when a competition factor of γ=1 was used, but growth of Salmonella was underestimated by ca. 1 log CFU/g. Unlike the salmon intermediate product, there was no obvious inactivation of pathogenic bacteria after the LAB starter culture reached its maximum population density. This was probably due to a higher pH of fermented blue marlin meat of above 5 compared to a pH of the fermented salmon mince below 5.
As there is no post-fermentation heat-treatment of the fermented blue marlin meat to inactivate potential pathogenic bacteria, the fermented blue marlin meat was assessed to have an insufficient safety margin to be used as a product or as an ingredient in other products processed without a post-fermentation heat-treatment.
Shelf-life tests of spreads
Since the intermediate product can be used for manifold product applications, the shelf-life of the final products is very much dependent on the further ingredients and the included processing steps. Therefore, main focus of the project work was on the safety aspects of the intermediate product. However, in order to get an indication for growth of spoilage organisms, shelf-life tests were exemplary carried out with the four different spreads produced during the pilot trials (Product 1: cold smoked; Product 2: cold smoked + salmon roe; Product 3: hot smoked; Product 4: hot smoked + trout roe).
Shelf-life studies were carried out with aerobically stored samples and in parallel with MAP-packaged (70% N2, 30% CO2) samples. These samples of each 2 x 100 g were stored at +7°C (± 0.5°) for 14 days.
At day 7, day 11 and day 14 following cell counts were determined:
a. Total cell count according ISO 4833
b. Enterobacteriaceae according ISO 21528
c. Lactic acid bacteria according ISO 15214
At all sampling points, no visible spoilage occurred. Basically, all cell counts were below levels which would indicate that the products are spoiled. After 14 days, all total cell counts were in a range between 1 x 105 cfu/g and 5 x 105 cfu/g, except of product 1 which had even slightly lower cell counts. No growth of any Gram-negative bacteria belonging to Enterobacteriaceae was determined. The cell counts on MRS (Cell densities of lactic acid bacteria) were almost in the same range as the total cell counts. This indicates that the spoilage flora is mostly dominated by lactic acid bacteria.
Based on these results it can be stated that a minimal shelf-life of at least 14 days conditions can be achieved under cooling, which allows a good marketing of the products.
More information as well as pictures of the products are documented in D4.2 and D5.1.
WP5, Pilot Validation and Quality Assurance
Task 5.1 Pilot testing of the FishFermPlus fermentation process
Pilot testing of the fermentation process developed in WP2 was performed at the plants of RAUCH and BG based on the pilot validation design developed in WP3. TTZ conducted the trials together with the SMEs and explained all relevant issues. This way, the SMEs got a clearer picture of practical aspects of the developed process. According to the developed sampling plan, samples were taken and analysed by TTZ. Results confirmed for both pilot trials findings from lab results. Thus, performed pilot trials indicate that the developed process is robust in an industrial surrounding, up-scalable, and reproducible.
Task 5.2 Pilot testing of the FishFermPlus product applications
Pilot testing of the product applications was performed in parallel with pilot testing of the fermentation process. Fermented products were directly used for pilot testing of the stabilization and product application steps. Due to some challenges that occurred during pilot testing at BG (see D5.1) the second process part was realized only at RAUCH. Ingredients developed in WP3 and WP4 were delivered by QFP. Results were identical with those achieved during lab trials as well. Produced products were used for development of heat inactivation models at DTU as well as for sensory evaluation at TTZ.
More detailed information about the pilot trials can be found in D5.1.
Task 5.3 Validation of shelf-life properties and consumer acceptance
Product applications from RAUCH were evaluated at TTZ in an objective and a subjective way. On the one hand, an expert panel described attributes and their intensities of the four different spreads. On the other hand, the same products were rated by consumers for their subjective liking. Correlations could be used to determine reasons for consumers’ preferences. All spreads were evaluated very positively and up to 90 % of the testers confirmed a buying intention. Thus, the results were a proof of successful process and product development and motivated RAUCH to follow the approach of spread development for later commercialization.
For later declaration BG and RAUCH desired the complete nutrient profile of the stabilized salmon mince. These will help for marketing activities e.g. by mentioning the good nutritional value due to unsaturated fatty acids and protein. According to food regulation, the declaration “contains omega-3-fatty acids” needs to be accompanied by the exact content of these fatty acids, which were part of the analysis. Furthermore, a product can be labelled as protein source if the energy from protein is at least 12 % of the products’ total energy value. For the salmon product the energy percentage from protein is 15 %.
For a more detailed knowledge about the safety and stability of the salmon intermediate product some chemical analyses were performed.
Low concentrations of putrescine, spermidine, spermine and tyramine were observed in salmon mince after fermentation, and other biogenic amines were below the limit of detection. The fermented salmon mince was mixed with different additives as described before and then heat treated. The concentrations of biogenic amines in the intermediate product were far below the concentrations that may cause potential health problems for human beings. Therefore, the final salmon intermediate product was assessed as safe with respect to biogenic amines.
Total volatile nitrogen (TVN), peroxide value (PV) and thiobarbituric acid reactive substance (TBARS) values of the salmon intermediate product stored at different temperatures were generally low. There were no significant differences (P>0.05) of TVN and TBARS values between frozen samples and samples storage at 5°C or 22.5°C, but storage temperatures reduced PV values of the samples significantly (P<0.05). The EU critical limit for TVN in salmon is 35 mg-N/100g (EC 2074/2005). The observed values of 14.7-16.8 mg-N/100g were much lower than the critical limit and in fact similar to concentrations typically observed for fresh fish. The fermented salmon intermediate product, therefore, did not show signs of microbiological spoilage. This was supported by sensory evaluation of the product where spoilage off-odours were not observed at the end of storage. There are no EU critical limits for PV and TBARS values corresponding to spoilage of seafood. The fact that PV and TBARS values did not increase during storage of the fermented and vacuum packed salmon product suggest that chemical changes during storage up to more than one month may not be the limiting factor for distribution of the product.
More details about the analyses can be found in D4.2 D5.1 and D5.2.
Task 5.4 Assessment of product safety & development of quality assurance methods
Safety assessment and shelf-life determination of the salmon intermediate product and the developed spreads, respectively, were performed as part of both WP4 and WP5. The work performed is described under task 4.2 and 5.3.
WP6 - Demonstration
Task 6.1 Planning of a demonstration event
As already mentioned in the DoW, the SMEs mentioned that organizing a demonstration event at their facilities would impact their daily business too much. Furthermore, they don’t have the capacities to host large events. Thus, it was decided to organize the external demonstration events alongside relevant exhibitions, where anyway a large amount of potentially interested parties is present.
For the internal demonstration events, SMEs agreed to combine them with the regular project meetings, in order to use time and resources as efficient as possible.
Task 6.2 Execution of the demonstration event
The first external demonstration event took place on 22-24 September 2014 at Seafood Expo Southern Europe in Barcelona. The SMEs identified this exhibition as relevant with regard to present exhibitors and visitors. Furthermore, the requirement from the DoW was met to hold one event in a country with a relevant fish market not represented in the project consortium. For maximum impact an own stand was organized where first pilot products were offered for tastings.
Since the SMEs highly appreciated this way of organizing an external demonstration event but no other relevant exhibition took place during the runtime of the project, it was decided to organize the second event at the Gastro Vision which is the most important sales exhibition for RAUCH and takes place only one month after project end.
The first internal demonstration event was held alongside the 4th project meeting. All parameters and processing steps had been fixed and were explained in detail to the SME partners. This allowed them to get a clearer picture of all potentials and requirements for implementation of the processes in their facilities. Discussions were a good basis for work to be done in WP3 and WP5 with regard to process integration and scale-up as well as pilot testing, respectively. Additionally, first product applications were tasted.
The second internal session was organized as part of the final project meeting. All project results were summarized and conclusions relevant for commercialization of process and products were given. Moreover, the general advantages of the FF+ process with regard to sustainability and economic aspects were highlighted.
Task 6.3 Demonstration monitoring, evaluation, and conclusions
At the Seafood Expo numerous visitors made use of the chance to taste the developed pilot products. Their feedback comprised mainly the nice sour note that was recognizable in the blue marlin products. This sourness is unusual for conventional fish products but was mentioned as very pleasant characteristic. Thus, the aim of creating new flavours by fermentation was positively received. For the salmon mince product the creamy texture was appreciated and also the flavor was well rated.
Due to the success of the first external demonstration event, the SMEs decided to organize the second event in the frame of an exhibition as well. The Gastro Vision as most important sales exhibition of RAUCH was identified as most relevant venue and was selected even though it will take place only in the month after project end. However, RAUCH will use the opportunity to present the newly developed spreads successfully tested in the consumer test to potential customers of the high quality sector. The feedback from this event will provide a more detailed information about market interest in and potential of FishFermPlus products and support further product optimization and identification of exploitation strategies during the post project period.
The internal demonstration events were also evaluated as very helpful by the SME partners. To see all relevant results and findings summarized in a condensed form helped them to get a clearer picture of the exact process requirements and opportunities. In this way, they could give a detailed feedback on their needs, expectations, and plans for later commercialization. Thus, an efficient collaboration and exchange between RTD and SME partners could be realized in addition to regular communication during development.
Conclusions for further commercialization and marketing activities are included in the PUDK (D8.4).
More details about the demonstration events are given in D6.1.
Potential Impact:
Delivering cost-efficient means for the processing and a larger valorization of yet underexploitedraw-materials to the European fish industry, and subsequently providing access toground-breaking physical processing and natural ingredients to increase shelf-life and safetyof fish and seafood products as proposed in the FishFermPlus approach will create withouta doubt a leap in competitiveness against other fish-exporting countries. Countless handicraftsto medium-scale SMEs from across the EU-27 working in fish and seafood products for b2b,retail, gastronomy, and end consumer will benefit from a direct economic impact, in addition toother SMEs providing respective equipment or compounds.
The benefits of the FishFermPlus results towards industry and innovative products create a win- /win- situation throughout the fish value chain which has considerable impact on consumers and society as well. Societal responsibilities are being addressed by a significant reduction in waste burdens – both in processing and retail – and by immense support given to the sustainable use of aquatic resources whilst delivering safe and high-quality convenience products to the European consumers.
The unique combination of addressing of key societal objectives next to the direct economic impact on the SME project beneficiaries as well as European SME competitiveness leads the consortium to believing that a broad exploitation of the FishFermPlus approach throughout the many countries with access to the sea or aquaculture farming and high activity in fish processing can be predicted. This will have an immense effect on safe-guarding and creating employment. To date, an exact figure cannot be forecasted on a reasonable basis other than multiplying the desirable market penetration with the number of fish-processors and an average additional staff head-count of 1-5 new employees; but it can be estimated to reach more than 1.000 upon market success.
A European-wide up-take of the proposed FishFermPlus technology will also reach consumers and society massively by its impact on improving the quality of life, health and safety: Fish and seafood account as a nutritious, healthy food on the one hand for its richness in omega-3 fatty acids (DHA, EPA) and for being a good source of high-quality protein in a well-balanced combination with vitamins and minerals. Increased fish consumption is thus frequently discussed in the light of preventing cardiovascular disease, inflammatory diseases or even Alzheimer’s. On the other hand, the safety of fish and seafood is equally challenged and questions as these foods are very sensitive and highly perishable. On this down-side, they can account for food poisoning by the accumulation and formation of undesirable substances (e.g. biogenic amines, residual contaminants) or the out- break of various food-borne diseases due to microbial spoilage associated mainly with mismanagement in the cold chain and or other sources of contamination when hygienic precautions are handled with too much levity. The FishFermPlus ingredients and product applications will be assessed against potential contents of undesirable components and will impact on the overall safety by competing and interacting directly with the growth of the product-specific microbial flora and/or specific contaminating strains which constitute main microbial risks (e.g. Listeria sp.) in fish and seafood. Thereby the product safety as well as the individual shelf-life will be improved. In addition to bringing additional safety and higher convenience with longer storage times and higher flexibility in the consumption, it is envisaged to use the FishFermPlus project approach also for a second type of fermentation strategy. It will target to add to the taste and texture properties of fish or even other savoury foods since these sensory criteria are very crucial to consumer acceptance. Frequently they are also the reason why the consumption of fish foods is declined or only reluctantly followed to fulfil the minimum nutrition recommendations. With taste and texture enhanced products, light users of fish and seafood may increase their consumption to reach preferable nutrition goals rather than a set minimum, whereas heavy users will appreciate the additional variety of high-quality foods found on the market. All types of consumers will further benefit from a new natural processing and preservation method, which is also in line with currently fast-growing consumer demands.
Finally, the concept of the FishFermPlus project seizes its role in society and environmental aspects in keeping with sustainable development for the fish industry and reducing the burden of waste discarding. This is achieved firstly by making use of high-quality, under-utilised raw materials (e.g. from fillet production or other trimmings, from mere optical failure from mechanical damage/strain) for the fermentation process. Currently more than 30% of the harvested fish and seafood gets classified as a by-product and thrown away during the processing stages of the fish value chain, not taking into account food products that remain uneaten, but get thrown away in the package due to the indicated end of shelf-life, especially in supermarkets where legal requirements have to be fulfilled in terms of only distributing safe foods. The FishFermPlus results address this context from two angles: (1) It is made possible for European fish processors to act and produce more resource- efficient, which contributes significantly to saving the aquatic resources. (2) Less fish products will be discarded after production/sales, because of the prolonged shelf-life and improved logistics. Whilst raising such issues and the respective company profiles of SMEs that seek proactive solutions for a better resource-management and waste reduction, it can also be envisaged that the general consumer awareness will be finally attracted towards socially responsible fish production and consumption then which is the ultimate goal for a number of current EC policies in the area of fishery and fish production. In this respect, again, not only the social and environmental aspects have to be considered, but also the policy objectives of the Seventh Framework Programme with regard to SMEs und their role in contributing to the ERA. With little R&D expenditure in this specific field of the agro- food sector, FishFermPlus will assist the EC in raising substantially the innovation level in the fish industry in keeping with the Lisbon Strategy.
Essential for a successful market uptake of products and processes developed within the FishFermPlus project is a well-thought plan for the dissemination of project results and for the exploitation thereof. The aim is to advertise the special characteristics and advantages of the developed products and processes and to encourage the uptake into current customers’ product portfolio as well as to broaden the customer range. In this way, it is ensured that SMEs’ investment into the project will pay off and that the project results contribute to enhancing their competitiveness.
In addition to commercial benefits for the fish industry, the project also aims to extend the scientific state-of-the-art knowledge with regard to fermentation of fish raw material and related impact on sensory and safety characteristics. The knowledge can be used to develop manifold new products with added value beyond the aims and applications of the FishFermPlus project.
Therefore, a series of demonstration and dissemination activities has been conducted during the lifetime of the project and isintended for the post project period. The target group comprises actors of fish manufacturing industry, fish suppliers, and agencies for fish industry (resellers), food retailers, food wholesalers, convenience food producers, high class gastronomy and mass caterers, food associations, food-related magazines as well as the scientific community. In order to sufficiently reach the envisaged target group and spread project information effectively, a diverse and versatile strategy has to be followed. The relevant parties were addressed and will continuously be contacted via personal communication and consultation, preparation of flyers providing information about the project consortium and the activities in the project as well as product properties, project website with information about the consortium and project objectives as well as news and online surveys, articles in relevant magazines, press releases and announcements on partners’ websites, presentations and distribution of information at exhibitions, trade fairs, and congresses related to fish production, food-industry in general, and biotechnology, production of a video clip promoting the project results as well as demonstration events.
Exploitable foreground generated within the project covers five main aspects, namely the pre-competitive FishFermPlus fermentation process, the FishFermPlus starter cultures/fermentates, the FishFermPlus stabilization ingredients, the fermented and stabilized FishFermPlus intermediate product, and the FishFermPlus pilot product applications.
Project beneficiaries BG Salmon and Die Räucherei will invest in further optimization of process flow and economic efficiency as well as in further product application development. Developed products will be offered to different food sectors. Envisaged time to market is one year. Developed starter cultures (fermentates) and stabilization ingredients are ready for the market and will be exclusively delivered by project beneficiary Quality Food & Products to BG Salmon and Die Räucherei during one year after the project, in order to guarantee a competitive advantage until the products are launched on the market. Afterwards they will be freely available on the market. Project beneficiary ISI Food Protection will use the knowledge generated within the project for further strategic research including transfer of the process to other products and applications.
List of Websites:
Website address: www.fishfermplus.eu
Project Consortium:
ISI Food Protection (Coordinator) www.isifoodprotection.com
Dieter Elsser-Gravesen deg@isifoodprotection.com
Quality Food & Products www.quality-food-products.de
Jürgen Oltmer info@quality-food-products.de
Die Räucherei www.dieraeucherei.de
Hans-Joachim Kunkel j.kunkel@die-raeucherei.com
BG Production www.bgsalmon.dk
Gisle Olsen go@bgsalmon.dk
ttz Bremerhaven www.ttz-bremerhaven.de
Annika Gering agering@ttz-bremerhaven.de
Technical University of Denmark www.food.dtu.dk
Paw Dalgaard pada@food.dtu.dk
In fish filet production almost one third of the high-value catch is lost, often sold as cheap animal feed. Aim of the FishFermPlus project has been to develop a process enabling more sustainable handling of scarce aquatic resources and a more economically efficient use for fish manufacturers. This is also of ecological and ethical interest in times of overfishing and of high protein demand of a growing world population.
Therefore, a bioprocess has been developed which allows the utilization of high-quality by-products from filet production. The process results in products with improved sensory characteristics on the one hand and increased safety and shelf-life on the other hand.
In order to achieve the envisaged objectives, the project consortium conducted an extensive literature review and identified by additional stakeholder consultations relevant market needs. The conclusion was that shelf-life prolongation is the most important goal for fish processors to increase quality and added value of their products.
In the following, suitable raw materials and cultures were tested to establish an effective bioprocess. In addition, optimal fermentation parameters were identified, which guarantee the production of value-added products produced in an economically efficient manner.
The fermented products were further processed, in order to obtain stable, marketable products. The stabilization steps included texturing processes, addition of antioxidants, salting as well as drying and heating treatments.
The stabilized intermediate products were finally incorporated in product applications. These comprised convenience products like spreads, burgers or filled puff pastry. However, it was shown that the products can also be used as ingredients e.g. as pizza toppings.
Safety assessment of the flexibly applicable intermediate product has shown that during the final heat treatment non-spore forming pathogens are efficiently killed, while spore-forming bacteria cannot grow during storage. Thus, with regard to food safety, the fermented and stabilized product was stable at room temperature (22.5 °C) during the observed storage time of 32 days.During chilled storage (5 °C) no pathogenic growth could be observed during 42 days. According to predictions including variability in product characteristics, contamination with Listeriaafter heating could lead to growth from 1 CFU/g to 100 CFU/g (critical limit) at 5 °C in 190 days with a lag time of 77 days. Considering the possibility of no lag time of Listeria and even larger variability in product characteristics, a safe shelf-life of 2 months was recommended.
Models have been developed which can be used to predict pH, lactic acid concentration as well as pathogenic growth during fermentation based on fermentation parameters. Models for heat inactivation are helpful to calculate effectiveness of different heating parameters with regard to reduction of pathogens. Further models are available to predict growth of Listeria during storage.
Sensory and chemical tests have shown that no off-flavours from fat oxidation occurred and that furthermore peroxide value (PV) and thiobarbituric acid reactive substance (TBARS) values did not increase during storage of the fermented and vacuum packed salmon product, suggesting that chemical changes during storage up to more than one month may not be the limiting factor for distribution of the product.
Pilot trials were performed in the facilities of the SME partners. Trials have indicated that upscaling of the process is realizable in an industrial setting. Furthermore, reproducible results compared to laboratory results were obtained. Thus, it is expected that further upscaling to industrial scale will be possible without remarkable challenges. Shelf-life tests of developed spreads have proven good product quality for at least two weeks.
During demonstration events the developed products were presented to potential customers. Furthermore, the opinion of the broad public was collected during a consumer test. A very positive feedback was received at these tastings which gives a promising indication for the products’ market potential.
During the post project phase further optimization with regard to final upscaling and industrialization of the developed process will be performed by the SME partners. Expected time to market of the products is one year after project end.
Project Context and Objectives:
The European fishing industry is one of the largest in the world. However, it faces strong competition from Asia and South America, with India and Russia catching up quickly. Despite a production of more than 5 Million tonnes of fish each year, the internal fish market tends to be not well-balanced: exports were made competition- and price-driven at a lower average price unit than imports for high-price raw materials.
This context shows clearly the need to create value in the internal market by serving for top quality demands, and by a clear distinction from cheap production markets. Such an approach requires real added-value with a tangible plus in terms of fish and seafood quality, safety and convenience. A mere price-/production unit-driven approach will not succeed for the European fish industry as the Union cannot compete with the production areas, high input aquaculture systems and coastlines of e.g. China, India and Russia, and has itself also committed to a more sustainable fish production with restrictions in catch and fleet quotas. Innovative alternative approaches are demanded since more than 350,000 jobs in the European fish industry are at risk, which are typically found in small- and medium-sized companies (SMEs) and account for almost 9% of the total European food industry, a major and top-priority pillar of European and rural economies.
Hence, addressing market mechanisms and consumer interests successfully will become a must to cope with future developments: Consumers were bombarded with news about foodrelated health crises around Europe which left them feeling insecure about the quality and safety of the foods they purchase. Changing lifestyles, work patterns, rising disposal income have resulted in more nutritional and health conscious consumers demanding top quality foods at utmost convenience for their use and storage. But also price remains an important driver in many European markets; and longer shelf- life is a key factor for high convenience and flexible supplies and has thus become indispensable.
Fish and seafood products can cater for many of these trends, but also face severe challenges from the latest developments: On the one hand, fish accounts for a “healthy food” fitting well into a balanced diet and delivering omega-3-fatty acids and other nutrients, which are considered to fight degenerative, age- or lifestyle-related diseases (Alzheimer’s, CVD etc.). On the other hand, it is a highly sensitive and perishable food, which requires accurate cold chain management and hygiene measures against food-borne diseases. It may even evoke fears of contaminants or other risk factors, such as sea pollutants, antibiotics etc. Analytical regimes and hygiene management systems are well developed and a number of technical singular solutions (e.g. packaging, preservatives, irradiation, canning, curing etc.) exist, but not all of them are evident to the consumer or well-appreciated as they do not match their demand for high-quality minimal processed foods, natural processing and ingredients, or waste minimisation. Instead guidance from simple symbols and reliable quality promises are sought-after as it can be observed from the MSC seal indicating sustainable production systems, or the Fresenius tick indicating compositional quality.
Next to the key challenge of the difficult product and market positioning to reach consumers, each fish processing company also faces very tangible day-to-day challenges arising from the operational and economic view-point of the business, which are again linked to a number of societal objectives and sustainable development as mainly the resource-efficiency in the fish processing is concerned. In the light of recent reports which point at the earlier (than previously estimated) arrival of the "fish dependence day", where Europe has nominally consumed all its own fish and needs to bring in stocks from elsewhere, it seems absurd that currently 30% of the initial catching are discarded and lost for human consumption along the total fish supply chain. Main factors accounting for this loss are frequently marketing requirements (portion sizes, trimmings, no visible mechanical damage of the fish flesh) towards a specific, intended use, and not food quality or safety concerns. The loss of resources and food wastage even continues beyond this figure in retail and consumer homes. Due to the short shelf-life and fear of food-borne diseases, restrictively set best-before-dates, and no checks of the actual food quality combined with inadequate stocking, many products are simply thrown away. The latter context may be basically out of the direct control of a fish producer, but affects his economic viability, if the shelf-life of his products is found to be too little to suit the retail and consumer needs.
Gaining a competitive edge in this context therefore means (a) to contribute against food wastage and towards sustainable development, (b) to fully utilise own resources optimally and (c) to provide real added-value products, which are distinct on the market in terms of food safety, quality and convenience/shelf-life. The latter cannot be achieved via chemical additives or extreme processing as done so and targeted in the past, because this will not achieve the crucial consumer acceptance and buying result on the market. The utilisation of available raw materials as suggested in (a) and (b) requires perspectives in addition to low-cost valorisation (e.g. minced sales, pressing).
The SME beneficiaries of the FishFermPlus project strongly believe in cost-efficient, simple, physical processing, such as fermentation, to deliver solutions against these needs. They see the potential in this approach to be inspired by cutting-edge biotechnological knowledge, but to deliver down-to-earth implementation solutions, which are broadly usable and clearly market driven. Hence, a balanced consortium of technology-driven SMEs supporting the food and fishindustry and of SME fish producers have gathered to outsource research activities to developand design such a process, but to remain with the full development power over the concertedprogram of activities in a Research for SMEs project.
The overall objective of the FishFermPlus project was the development of a bioprocess which generates new properties by treatment of fish raw material and by-products of fish filet production. In this way, an added value has been created, by improved sensory characteristics as well as by enhanced safety and shelf-life. In the end, additional economic value will be created for fish-processing companies by valorisation of underutilized material and food wastage can be reduced.
In order to succeed with this overall mission, the FishFermPlus project needed to fulfil a number of technical and operational objectives:
(1) A clear definition and specifications of the actual new process design had to be achieved by a bottom-up approach for the compilation of detailed requirements. These had to be evaluated positively by the end-users included in the FishFermPlus consortium and also in the frame of confidential in-depth consultations with representatives from the fish (processing) industry. Despite having gained insight into technological needs, opportunities, cost restraints and actual real business capacities by various methods, such as market reviews, online surveys, literature reviews and in persona consultations, a conclusive blueprint for the development was elaborated.
(2) The identification of suitable strains, cultures, substrates and other ingredient components to promote desirable microbial growth and the production of target metabolites was most crucial for the whole project success. For this purpose a highly efficient screening method was implemented to maintain development speed and result-orientation in the identification process, the selection and exploration of suitable combinations, as well as the latter optimisation.
(3) Once a viable and efficient microbiological system had been developed for the fermentation of fish substrates, finding an optimal balance for the stabilisation of the FishFermPlus ingredients was required in order to arrive at industrially applicable products. Under this objective, the need for sensitive and gentle product processing (to maintain functionality, sensory properties and overall shelf-life of the ingredients) had to be addressed as well as the final costs and operability of the process.
(4) The next crucial operational objective revolved around the detailed exploration of the targeted ingredients/compounds/intermediate products and an in-depth investigation of their effect on the manufacturing of new fish and seafood products. It was envisaged to arrive at a minimum number of 3 added-value application processes, which will have significant economic impact for the end-user SMEs. To this end, it was foreseen to achieve and to document the development and validation of re-texturized products to be used as spreads, fillings, toppings or bratwurst-like sausages. The performance of the ingredients/compounds/intermediate products and the actual characteristics in terms of antimicrobial, flavour-/texture-improving activity or prolongation of shelf-life needed to be verified and validated at various development stages.
(5) As market-oriented project, FishFermPlus further aimed at contributing to underpinning quality assurance measures by evaluation of models for product changes based on data obtained in pilot validations. Such models will be an important building block towards the future marketing of the new FishFermPlus products and could potentially be used in terms of technology transfer to new customer companies. A specific task has been dedicated for assessing all data and performing safety assessments by applying relevant modelling approaches.
(6) Physically, FishFermPlus aims at delivering a bioprocessing system to the market. Hence, crucial technological and operative objectives concern also the integration of potential batch sizes, desirable production times, scheduling and logistics, energy costs etc. in order to enable the latter broad implementation and exploitation. It is expected that a latter investment in the bioprocess by future customer companies should not exceed € 30.000 in order to promote the up-take. Next to the measurable cost factor, the functionality of the system will also be fully assessed by testing and validation of the FishFermPlus technology in real environments with products of market concern and value, which will deliver documentation for the FishFermPlus added-value beyond the scope of scientific small-scale trials. The scope of the project enfolded technological requirements and benefits, as well as considering socioeconomic factors.
(7) The final operational objective relates to the necessary pre-requisites for realising the targeted impact and return-on-investment by a broad support of the up-take of the FishFermPlus results by SMEs and a wider audience. The successful accomplishment was and will be supported by carrying out a comprehensive series of demonstration and training activities which will prove the viability and outline the economic potential due to real product quality advantages.
Project Results:
WP 1 - Stakeholder Targeting and Definition of Industrial Specifications
Task 1.1 European-wide market review and in-depth consultations
A short questionnaire was developed by DTU in order to identify a representative opinion of the fish sector regarding the FishFermPlus objectives of shelf-life prolongation and sensory improvement of fish products. The questionnaire was available online and was translated by DTU into French and by TTZ into Spanish, in order to improve the response of the Mediterranean fish sector, which is not represented in the project consortium. In addition to addressing concrete companies, also associations were contacted by DTU and TTZ as multipliers which were asked to forward the questionnaires to their members. In total, 170 Spanish, 45 French as well as 393 further European stakeholders were contacted and asked to answer the questionnaire. The response rate was quite poor, only 15 questionnaires were filled. As experienced by the beneficiaries in other projects, it is generally hard to convince companies to take part in such a survey, due to the fact that they are too busy with their daily business and are afraid of revealing confidential business information.
The analysis of the collected responses results in the following prioritized list of needs and expectations:
1) Antimicrobial properties of a fermented ingredient are important (High importance: 86 % of responses) to obtain a shelf-life extension of 10-50%.
2) Texture improving properties of a fermented ingredient are important (High importance: 57 %) to reduce liquid loss and to facilitate slicing (and reduce slicing losses).
3) Flavour enhancing properties of a fermented ingredient are important (High importance: 43 %) primarily to reduce off-odour and off-flavour development.
4) A fermented ingredient is interesting even if it has to be declared e.g. as ‘protective culture’ (71.4%).
The exact results of the market review can be found in D1.1.
In addition to addressing the European industry, extended questionnaires were sent to the SME partners ISI, QFP, RAUCH, and BG. These were accompanied by personal consultations from DTU and TTZ in order to obtain more information about the SMEs’ interests regarding specific raw material and properties they would be interested in. This information was also used for identification of processing and application specifications in tasks 1.4 and 1.5.
In addition to the results from the market review, the interviews with the project partners revealed that for low-price products, like those of BG, shelf-life extension is of most importance. Whereas for high-price products and especially for products targeted to the catering and gastronomy sector, which do not need a long shelf-life, the sensory properties are more relevant.
Task 1.2 Literature research
An intense literature review of latest scientific findings in relation to fish fermentation was performed by DTU. As discussed with the whole consortium at the kick-off meeting, this was mainly focused on the traditional Asian market, since it is the largest market for fermented fish products worldwide. The report contains information about types of fermented fish, microorganisms detected in fermented fish as well as their properties, texture and flavour compounds in fermented fish, shelf-life and food safety issues of fermented fish, biopreservation methods as well as commercial cultures. However, hardly any studies could be found with regard to controlled, standardized fermentation, which underlines the high innovative approach of the FishFermPlus project.
The whole literature review can be found in D1.2.
Task 1.3 National and international standards and legislation requirements
TTZ supported by DTU compiled an overview of legislations and standards of relevance for the FishFermPlus project. Support came from the partners RAUCH and BG who reported about requirements in their productions. The report contains aspects about conditions for using living cultures in food, microbiological criteria in food, especially concerning listeria and histamine, bacteriocins in food, labelling requirements, novel food regulations as well as the Good Manufacturing Practices.
This comprises EFSA’s QPS-list of cultures considered as safe, the inventory of microorganisms with a documented history of use in food published by the International Dairy Federation and the European Food and Feed Cultures Association, Commission Regulation (EC) No 1441/2007 on microbiological criteria for foodstuffs containing limits for L. monocytogenes and histamines in fish, labelling requirements according to Directive 2000/13/EC as well as current Good Manufacturing Practices (GMP) published by the US Food and Drug Administration with special focus on microbial food enzyme production.
The results are presented together with the literature review in D1.2.
As demanded in the DoW, fish sector associations were contacted by DTU and TTZ in the frame of task 1.2. However, for the legislation report, the aspects to consider were focussed on the work to be performed and developments planned during the project rather than on the general standards implemented in industry.
Task 1.4 FishFermPlus processing requirements & specifications and Task 1.5 FishFermPlus application and end-user requirements & specifications
As described in D1.3 SME partners found it difficult to define exact processing and application specifications, since especially the pricing depends on the real outcome and benefits achieved during the practical trials. The final type of the fermented products is not clear yet either, as final stabilization methods still need to be tested. Thus, concrete threshold characteristics like pH and aw-values are hard to define at the current stage of the project.
However, the SME partners ISI, QFP, RAUCH, and BG supported by the RTDs TTZ and DTU worked out possible strategies that could lead to products of interest. The concept comprises two approaches. One attempt is tailored for high-quality raw material like filets as well as bits and pieces and aims primarily at the ripening and creation of new sensory characteristics. On the other hand, low-value fish mince will be fermented to achieve mainly shelf-life and safety improving properties. Both approaches are described in D1.3 and are the basis for practical trials in WP2.
WP2 - Development of the fermentation process
Task 2.1 Experimental investigation design and overall development approach
For development of the fermentation design numerous trials were conducted (D2.2). For the fermentation repository it proved successful to ferment in plastic bags with controlled vacuum. For blue marlin a vacuum of 70 % was identified as optimal parameter whereas salmon mince was fermented at 25 % vacuum.
Selection of fermentation substrates, starter cultures as well as fermentation parameters is described below.
Control parameters to compare successful fermentation and reproducibility of the process were pH of the raw material, inoculation cell count as well as pH, LAB cell count, and smell after fermentation.
Task 2.2 Substrate selection
Minced salmon was fixed as substrate for fermentation since it is the main by-product from filet production at BG’s production process.
Since tuna proved unsuitable for fermentation in first trials, further fish species of high relevance in the business of RAUCH were selected, namely wild Alaska salmon and blue marlin. Finally, blue marlin was selected due to better sensory results during fermentation.
Task 2.3 Strain identification and culture development
Selection of strains to be tested was done jointly by TTZ, DTU, QFP, and ISI according to results from the literature review in WP1 as well as based on experience by ISI and QFP. Only food-grade cultures were selected. In total, 26bacteria for acidification as well as seven yeasts for flavour development were tested.
Strains were provided by ISI or bought from the German Collection of Microorganisms and Cell Cultures (DSMZ) and Chr. Hansen. All cultures were included in TTZ’s strain collection and were cultivated by TTZ.
In addition to single strains also fermented food products were used as fermentates in order to test their influence on flavour production. The following products were used:
- Sauerkraut juice
- Bread drink
- Kombucha
- Water kefir
- Natural yoghurt
Identification of final starter cultures is described under task 2.5.
Task 2.4 Final selection of processes and raw materials
For the blue marlin bits and pieces, L. diacetylactis was identified as most effective flavour producing strain. The developed butter flavour was clearly recognizable and evaluated by RAUCH as interesting new product characteristic that will add value to the bits and pieces and create a unique selling proposition of later products. Combination with 0.25 % ascorbic acid was most effective in intensifying the butter flavour. Further parameters defined include 2 % glucose, 0.005 % booster, 20 h fermentation time, fermentation in evacuated bags with a vacuum of 70 % and a fermentation temperature of 25 °C.
For the salmon mince, L. diacetylactis could produce a pleasant flavour as well. Flavour characteristics could be improved by 10 minutes cooking prior fermentation and by addition of 0.25 % citric acid. Addition of 2 % glucose, 0.005 % booster, 20 h fermentation time, fermentation in evacuated bags with a vacuum of 25 % and a fermentation temperature of 25 °C proved effective also in this case.
Task 2.5 Process optimisation and definition of parameters for transfer
During optimization trials different strains of L. diacetylactis were investigated. Thereby, different results were achieved for blue marlin and salmon mince with regard to sensory characteristics and intensity of the produced butter flavour. Therefore, two individual starter cultures were selected, DSM 20384 for blue marlin as well as DSM 20729 for salmon mince. During stabilization trials in WP3 these were produced as freeze-dried cultures for easier handling during upscaling trials and use in an industrial environment in WP3 and WP5.
Furthermore, for reproducibility of pleasant sensory results it was decided to ferment with L. diacetylactis alone and not in combination with other strains as indicated in D2.2.
During reproducibility and validation trials the impact of raw material quality became evident. Problems with blue marlin quality on the world market had a negative influence of sensory results after fermentation. Therefore, the optimization development became delayed with regard to original project schedule. For later commercialization it is highly recommended to ensure raw material quality for reproducible fermentation results. Further aspects that led to a delay in fermentation optimization due to insufficient reproducibility results included the need for better standardization of processing steps and their adaptation by both RTD partners.
For salmon mince it was obvious that the material was highly perishable due to its large surface and the high oil content with regard both to microbial spoilage and fat oxidation. Optimization and standardization of the raw material quality could be achieved by direct evacuation and freezing after production.
Calculated inoculation concentration was slightly increased from 106 cfu/g to 3x106 cfu/g, since trials had shown that actual cell count in the product was always a bit lower.
WP3 - Downstream Processing & Equipment Design
Task 3.1 Selection and optimization of stabilization technologies
The original objective was to identify stabilization processes for the fermented products in order to obtain stable and functional fermentates for later compound development and use of these in different product applications.
However, drying tests in WP3 have shown that due to the high oil content of the salmon mince from BG it was very challenging to obtain a powdery product. Even with high percentage (up to 80 %) of the carrier substance maltodextrin the result was not satisfying. Salmon mince is the major material occurring in high amounts at BG and there is no other material of interest for them to replace the salmon mince. Therefore, the partners agreed not to continue the approach of drying and compound development, since the development of further procedures would be too time- and resource-consuming. Furthermore, the resulting process was likely to be too sophisticated and expensive to be feasible and efficient for later industrial implementation. Also the oil removal was not favoured, since it would mean an additional processing step. At the same time, the aim of utilizing the material completely would not be reached.
Therefore, the original goal of drying for further use as fermentates had to be modified as part of the DoW amendment n°1. While the developed starter cultures are available to be offered as fermentates, main new task was the stabilization of the fermented products for further development to intermediate products and related product applications. Thereby, the restructuring of the bits and pieces as well as of the salmon mince became the main focus together with avoidance of fat oxidation.
Since trials revealed that fermented blue marlin pieces were too instable to glue them with economically efficient concentrations of transglutaminase. Therefore, it was decided to ferment whole filets which can be sold as high quality products, which even increased value on the one side. On the other side, bits and pieces should be fermented as cubes which can be sold e.g. as salad toppings.
Drying trials were started with whole filets. Tests were performed directly in RAUCH’s facilities in order to get realistic results with their equipment and conditions. Furthermore, this was more efficient with regard to later process integration and scale-up. Identification of optimal parameters was mostly based on RAUCH’s experience in other product applications. Trials revealed that drying of products without addition of smoke was more promising, since sensory evaluation indicated that the subtle fermentation flavour was completely covered by the smoke aroma. The selected processing parameters comprised a ripening step with salt and additional spices for 48 h at 2-4 °C followed by a drying step for 7.5 h at 26 °C at a humidity of 30 %.
For salmon mince a process was established that transferred the inhomogeneous mass into a smooth, spreadable product. Therefore, an emulsification process was developed that included addition of oil, water as well as a mixture of stabilizers.
Since BG expressed also interest in a sliceable texture, further trials were done with modified ingredient concentrations. However, the result showed that due to the denaturation of proteins during the cooking process prior fermentation, the bonding of proteins was hampered. For maximum benefit of BG and later utilization of their salmon mince, further trials were conducted with unfermented mince. With the uncooked material a sliceable bologna-like texture could be achieved with the developed stabilizing agents. This will offer advantages for different applications, although without the special benefits targeted at by fermentation like improved safety and shelf-life.
Another challenge of the salmon mince resulting from the high fat content was the product’s susceptibility to fat oxidation and related unpleasant sensory rancidity notes. QFP has developed a compound made of grinded rosemary which is known for its antioxidant capacity.
Task 3.2 Process integration and scale-up
In preparation for the pilot validation, first up-scaling tests for the salmon mince fermentation developed in WP2 were conducted at TTZ’s facilities. Heating in a heating reactor with stirrer was performed with 7 kg of salmon mince raw material. For mixing of fermentation ingredients a cutter was used with the same capacity. Incubation was done in 1 kg bags.
Based on developments from task 3.1 and WP4, upscaling trials for salmon mince stabilization and product application were performed as well. A batch size of 5 kg was processed in a HydroGrind allowing high shear forces with rotations of 90 Hz. The resulting products showed a more homogenous emulsion compared to lab trials by QFP as well as stable properties after freezing for several weeks and subsequent defrosting.
Results confirmed those achieved at lab scale with regard to growth of starter culture and pH decrease. Thus, first scale-up trials indicated an easy upscalable process.
Products produced during the upscaling trials were used for the first external demonstration event at the project’s Seafood Expo stand (see WP6) as well as for safety assessment trials by DTU in WP4 and 5.
During the planning of the pilot trials, safety assessment by DTU revealed that the fermented blue marlin is not sufficiently safe. The development of an additional process for stabilization of blue marlin was not feasible within the remaining time. Therefore, it was decided not to perform pilot validation of blue marlin. Instead, RAUCH expressed the interest to test salmon mince processing in their plant as well.
More details are described in D3.1.
Task 3.3 Pilot validation design and building
After successful first upscaling tests, next step was the transfer of the whole process to pilot scale in the plants of the producer SMEs RAUCH and BG. As described above, it was decided not to perform pilot validation with blue marlin, as initially planned. Therefore, pilot validation of the salmon process was planned both for RAUCH’s and BG’s facilities. Pilot validation design was the basis for trials to be performed in WP5.
Due to extensive discussions between the SMEs and TTZ it was guaranteed that all chosen equipment was appropriate for a promising conduction of pilot trials and reproducibility of lab-scale results. Due to sufficient actual testing environment, it was not necessary to assemble any additional equipment. Additionally, no specific adaptions of available equipment were needed.
Aim of the pilot trials was to validate that the results achieved in realistic industrial surrounding are in accordance with laboratory results. Therefore, an analysis protocol was developed for recording pH values, cell count, heating profiles as well as smell.
More information about the selected equipment is given in D3.2.
WP4 - Development of Product Applications
Task 4.1 FishFermPlus intermediate products development
Stabilized salmon mince developed in WP3 were used as basis for further development of the intermediate product. As a result, a fish flavour ingredient has been developed containing a smoke substance for support of antimicrobial activity and spices that harmonize fish the aroma from the fermented salmon and the rosemary flavour from the Antiox applied during stabilization. A process was established for combined application of stabilization and flavour ingredients. The resulting product can be used either directly as a spread or as an ingredient in other product applications. All market needs identified in WP1 including texture improving properties, flavour enhancing properties, and a fermentate for application in fish processing could be met. The last aspect of shelf-life extension is described below. In addition, price calculation revealed that the developed product can be marketed at a profitable and competitive price.
Task 4.2 Product application development and validation
The intermediate product developed in task 4.1 was further developed for several product applications. Since RAUCH is selling in the premium catering sector focussing mainly on unique products of high quality, while BG distributes products in a lower price sector, different product application were developed with both partners.
For BG main interest is to sell the intermediate product as ingredient for convenience products like salads, pizza, puff pastry etc. For using it as slices on pizza or cubes on salads a sliceable texture was desired by BG. As described in D3.1 this could not be achieved completely. However, trials performed by QFP in cooperation with TTZ and BG revealed that the texture was solid enough for the mentioned applications. In addition, several convenience applications were tested and proven feasible like fish hamburgers, fish nuggets, and salmon filled pastry. The intermediate product could be shown to be stable in texture during baking and frying.
Due to the challenges that occurred during texturing of blue marlin pieces (see D3.1) it was decided to ferment whole blue marlin filets, in order to allow applications for RAUCH’s desired raw material and to be able to present products at the already planned Seafood Expo. RAUCH developed three types of seasoning, namely natural, sesame, and pastrami, which matched and did not mask the fermentation flavour.
Due to the results from safety assessment, that revealed that fermented blue marlin is not safe enough, RAUCH decided to develop product applications for the salmon mince as well. Since RAUCH delivers products mainly to the catering and gastronomy sector, they developed a convenience product e.g. for bakery to-go products. In total, they developed four spreads containing next to the FishFermPlus product, smoked salmon cream, mayonnaise and partly roe. These products were used for pilot production and consumer evaluation in WP5.
Safety assessment of intermediate salmon product
Microbial safety of the fermented salmon mince product was assessed. The fermentation process was well characterized including growth of added starter culture (lactic acid bacteria), change of pH and formation of lactic acid during fermentation.
Challenge tests were performed for three key steps including fermentation, heating to improve product texture and final chilled storage. Growth and survival of seven pathogenic bacteria, including Bacillus cereus, non-proteolytic Clostridium botulinum, E. coli, Listeria monocytogenes, Salmonella, Staphylococcus aureus and Yersinia enterocolitica were assessed. Concentrations of pathogenic bacteria increased 2 to 4 log CFU/g during fermentation at 25°C except for non-proteolytic Cl. botulinum where 1 to 2 log-reductions was observed. Starter lactic acid bacteria reduced but did not prevented growth of pathogenic bacteria during fermentation at 25°C. Heating of fermented salmon mince reduced concentrations of the studied non-spore-forming pathogenic bacteria by more than 6 log CFU/g and of spore-formers by 1 to 2 log CFU/g. This heating step was important for safety of the product (3.3 in the Appendix). None of the evaluated pathogenic bacteria were able to grow in the fermented product at 5 °C but at 22.5 °C, representing room temperature, growth was exclusively observed for L. monocytogenes, Salmonella and St. aureus. The fermented and textured salmon mince product was found to be sufficiently preserved to allow distribution under chilled and room temperature conditions when the product was heat treated in the final and sealed packaging used for distribution. If the product was processed including repacking after the heat treatment then contamination and subsequent growth of L. monocytogenes, St. aureus and Salmonella were identified as risk with respect to safety of the fermented salmon mince product. Observed growth or survival data were compared with predictions from available models and in general these models had poor performance. New models were developed for growth of lactic acid bacteria and related lactic acid formation and pH reduction during fermentation as well as for growth of L. monocytogenes during storage of the final product. These models were used to evaluate the effect of variability in product characteristics on safe shelf-life of the fermented salmon mince product. By using L. monocytogenes as safety indicator bacteria, and evaluation of variability in product characteristics of 6 samples from different batches, the safe shelf-life of the salmon intermediate product could be determined as 76 days at 5 °C from assistance of the model.
Safety assessment of blue marlin during fermentation
Challenge tests for microbiological safety of fermented blue marlin meat were performed. Growth and survival of three pathogenic bacteria, including L. monocytogenes, Salmonella and St. aureus were assessed. The methods used for challenge testing of fermented blue marlin meat were similar to those used for fermented salmon mince.
The fermentation process at 25°C resulted in unacceptable growth of all three investigated pathogenic bacteria of 2.5 to 4.0 log CFU/g. Starter lactic acid bacteria (LAB) reduced but did not prevented growth of the pathogenic bacteria during fermentation at 25°C. Growth and interaction for starter LAB and both Listeria monocytogenes and Staphylococcus aureus seemed to correspond to the Jameson effect. The observed growth/survival of starter LAB and the three pathogenic bacteria was compared with predictions by models developed for the salmon intermediate product. Models for the salmon intermediate product provided relatively precise predictions for growth of L. monocytogenes and St. aureus during fermentation when a competition factor of γ=1 was used, but growth of Salmonella was underestimated by ca. 1 log CFU/g. Unlike the salmon intermediate product, there was no obvious inactivation of pathogenic bacteria after the LAB starter culture reached its maximum population density. This was probably due to a higher pH of fermented blue marlin meat of above 5 compared to a pH of the fermented salmon mince below 5.
As there is no post-fermentation heat-treatment of the fermented blue marlin meat to inactivate potential pathogenic bacteria, the fermented blue marlin meat was assessed to have an insufficient safety margin to be used as a product or as an ingredient in other products processed without a post-fermentation heat-treatment.
Shelf-life tests of spreads
Since the intermediate product can be used for manifold product applications, the shelf-life of the final products is very much dependent on the further ingredients and the included processing steps. Therefore, main focus of the project work was on the safety aspects of the intermediate product. However, in order to get an indication for growth of spoilage organisms, shelf-life tests were exemplary carried out with the four different spreads produced during the pilot trials (Product 1: cold smoked; Product 2: cold smoked + salmon roe; Product 3: hot smoked; Product 4: hot smoked + trout roe).
Shelf-life studies were carried out with aerobically stored samples and in parallel with MAP-packaged (70% N2, 30% CO2) samples. These samples of each 2 x 100 g were stored at +7°C (± 0.5°) for 14 days.
At day 7, day 11 and day 14 following cell counts were determined:
a. Total cell count according ISO 4833
b. Enterobacteriaceae according ISO 21528
c. Lactic acid bacteria according ISO 15214
At all sampling points, no visible spoilage occurred. Basically, all cell counts were below levels which would indicate that the products are spoiled. After 14 days, all total cell counts were in a range between 1 x 105 cfu/g and 5 x 105 cfu/g, except of product 1 which had even slightly lower cell counts. No growth of any Gram-negative bacteria belonging to Enterobacteriaceae was determined. The cell counts on MRS (Cell densities of lactic acid bacteria) were almost in the same range as the total cell counts. This indicates that the spoilage flora is mostly dominated by lactic acid bacteria.
Based on these results it can be stated that a minimal shelf-life of at least 14 days conditions can be achieved under cooling, which allows a good marketing of the products.
More information as well as pictures of the products are documented in D4.2 and D5.1.
WP5, Pilot Validation and Quality Assurance
Task 5.1 Pilot testing of the FishFermPlus fermentation process
Pilot testing of the fermentation process developed in WP2 was performed at the plants of RAUCH and BG based on the pilot validation design developed in WP3. TTZ conducted the trials together with the SMEs and explained all relevant issues. This way, the SMEs got a clearer picture of practical aspects of the developed process. According to the developed sampling plan, samples were taken and analysed by TTZ. Results confirmed for both pilot trials findings from lab results. Thus, performed pilot trials indicate that the developed process is robust in an industrial surrounding, up-scalable, and reproducible.
Task 5.2 Pilot testing of the FishFermPlus product applications
Pilot testing of the product applications was performed in parallel with pilot testing of the fermentation process. Fermented products were directly used for pilot testing of the stabilization and product application steps. Due to some challenges that occurred during pilot testing at BG (see D5.1) the second process part was realized only at RAUCH. Ingredients developed in WP3 and WP4 were delivered by QFP. Results were identical with those achieved during lab trials as well. Produced products were used for development of heat inactivation models at DTU as well as for sensory evaluation at TTZ.
More detailed information about the pilot trials can be found in D5.1.
Task 5.3 Validation of shelf-life properties and consumer acceptance
Product applications from RAUCH were evaluated at TTZ in an objective and a subjective way. On the one hand, an expert panel described attributes and their intensities of the four different spreads. On the other hand, the same products were rated by consumers for their subjective liking. Correlations could be used to determine reasons for consumers’ preferences. All spreads were evaluated very positively and up to 90 % of the testers confirmed a buying intention. Thus, the results were a proof of successful process and product development and motivated RAUCH to follow the approach of spread development for later commercialization.
For later declaration BG and RAUCH desired the complete nutrient profile of the stabilized salmon mince. These will help for marketing activities e.g. by mentioning the good nutritional value due to unsaturated fatty acids and protein. According to food regulation, the declaration “contains omega-3-fatty acids” needs to be accompanied by the exact content of these fatty acids, which were part of the analysis. Furthermore, a product can be labelled as protein source if the energy from protein is at least 12 % of the products’ total energy value. For the salmon product the energy percentage from protein is 15 %.
For a more detailed knowledge about the safety and stability of the salmon intermediate product some chemical analyses were performed.
Low concentrations of putrescine, spermidine, spermine and tyramine were observed in salmon mince after fermentation, and other biogenic amines were below the limit of detection. The fermented salmon mince was mixed with different additives as described before and then heat treated. The concentrations of biogenic amines in the intermediate product were far below the concentrations that may cause potential health problems for human beings. Therefore, the final salmon intermediate product was assessed as safe with respect to biogenic amines.
Total volatile nitrogen (TVN), peroxide value (PV) and thiobarbituric acid reactive substance (TBARS) values of the salmon intermediate product stored at different temperatures were generally low. There were no significant differences (P>0.05) of TVN and TBARS values between frozen samples and samples storage at 5°C or 22.5°C, but storage temperatures reduced PV values of the samples significantly (P<0.05). The EU critical limit for TVN in salmon is 35 mg-N/100g (EC 2074/2005). The observed values of 14.7-16.8 mg-N/100g were much lower than the critical limit and in fact similar to concentrations typically observed for fresh fish. The fermented salmon intermediate product, therefore, did not show signs of microbiological spoilage. This was supported by sensory evaluation of the product where spoilage off-odours were not observed at the end of storage. There are no EU critical limits for PV and TBARS values corresponding to spoilage of seafood. The fact that PV and TBARS values did not increase during storage of the fermented and vacuum packed salmon product suggest that chemical changes during storage up to more than one month may not be the limiting factor for distribution of the product.
More details about the analyses can be found in D4.2 D5.1 and D5.2.
Task 5.4 Assessment of product safety & development of quality assurance methods
Safety assessment and shelf-life determination of the salmon intermediate product and the developed spreads, respectively, were performed as part of both WP4 and WP5. The work performed is described under task 4.2 and 5.3.
WP6 - Demonstration
Task 6.1 Planning of a demonstration event
As already mentioned in the DoW, the SMEs mentioned that organizing a demonstration event at their facilities would impact their daily business too much. Furthermore, they don’t have the capacities to host large events. Thus, it was decided to organize the external demonstration events alongside relevant exhibitions, where anyway a large amount of potentially interested parties is present.
For the internal demonstration events, SMEs agreed to combine them with the regular project meetings, in order to use time and resources as efficient as possible.
Task 6.2 Execution of the demonstration event
The first external demonstration event took place on 22-24 September 2014 at Seafood Expo Southern Europe in Barcelona. The SMEs identified this exhibition as relevant with regard to present exhibitors and visitors. Furthermore, the requirement from the DoW was met to hold one event in a country with a relevant fish market not represented in the project consortium. For maximum impact an own stand was organized where first pilot products were offered for tastings.
Since the SMEs highly appreciated this way of organizing an external demonstration event but no other relevant exhibition took place during the runtime of the project, it was decided to organize the second event at the Gastro Vision which is the most important sales exhibition for RAUCH and takes place only one month after project end.
The first internal demonstration event was held alongside the 4th project meeting. All parameters and processing steps had been fixed and were explained in detail to the SME partners. This allowed them to get a clearer picture of all potentials and requirements for implementation of the processes in their facilities. Discussions were a good basis for work to be done in WP3 and WP5 with regard to process integration and scale-up as well as pilot testing, respectively. Additionally, first product applications were tasted.
The second internal session was organized as part of the final project meeting. All project results were summarized and conclusions relevant for commercialization of process and products were given. Moreover, the general advantages of the FF+ process with regard to sustainability and economic aspects were highlighted.
Task 6.3 Demonstration monitoring, evaluation, and conclusions
At the Seafood Expo numerous visitors made use of the chance to taste the developed pilot products. Their feedback comprised mainly the nice sour note that was recognizable in the blue marlin products. This sourness is unusual for conventional fish products but was mentioned as very pleasant characteristic. Thus, the aim of creating new flavours by fermentation was positively received. For the salmon mince product the creamy texture was appreciated and also the flavor was well rated.
Due to the success of the first external demonstration event, the SMEs decided to organize the second event in the frame of an exhibition as well. The Gastro Vision as most important sales exhibition of RAUCH was identified as most relevant venue and was selected even though it will take place only in the month after project end. However, RAUCH will use the opportunity to present the newly developed spreads successfully tested in the consumer test to potential customers of the high quality sector. The feedback from this event will provide a more detailed information about market interest in and potential of FishFermPlus products and support further product optimization and identification of exploitation strategies during the post project period.
The internal demonstration events were also evaluated as very helpful by the SME partners. To see all relevant results and findings summarized in a condensed form helped them to get a clearer picture of the exact process requirements and opportunities. In this way, they could give a detailed feedback on their needs, expectations, and plans for later commercialization. Thus, an efficient collaboration and exchange between RTD and SME partners could be realized in addition to regular communication during development.
Conclusions for further commercialization and marketing activities are included in the PUDK (D8.4).
More details about the demonstration events are given in D6.1.
Potential Impact:
Delivering cost-efficient means for the processing and a larger valorization of yet underexploitedraw-materials to the European fish industry, and subsequently providing access toground-breaking physical processing and natural ingredients to increase shelf-life and safetyof fish and seafood products as proposed in the FishFermPlus approach will create withouta doubt a leap in competitiveness against other fish-exporting countries. Countless handicraftsto medium-scale SMEs from across the EU-27 working in fish and seafood products for b2b,retail, gastronomy, and end consumer will benefit from a direct economic impact, in addition toother SMEs providing respective equipment or compounds.
The benefits of the FishFermPlus results towards industry and innovative products create a win- /win- situation throughout the fish value chain which has considerable impact on consumers and society as well. Societal responsibilities are being addressed by a significant reduction in waste burdens – both in processing and retail – and by immense support given to the sustainable use of aquatic resources whilst delivering safe and high-quality convenience products to the European consumers.
The unique combination of addressing of key societal objectives next to the direct economic impact on the SME project beneficiaries as well as European SME competitiveness leads the consortium to believing that a broad exploitation of the FishFermPlus approach throughout the many countries with access to the sea or aquaculture farming and high activity in fish processing can be predicted. This will have an immense effect on safe-guarding and creating employment. To date, an exact figure cannot be forecasted on a reasonable basis other than multiplying the desirable market penetration with the number of fish-processors and an average additional staff head-count of 1-5 new employees; but it can be estimated to reach more than 1.000 upon market success.
A European-wide up-take of the proposed FishFermPlus technology will also reach consumers and society massively by its impact on improving the quality of life, health and safety: Fish and seafood account as a nutritious, healthy food on the one hand for its richness in omega-3 fatty acids (DHA, EPA) and for being a good source of high-quality protein in a well-balanced combination with vitamins and minerals. Increased fish consumption is thus frequently discussed in the light of preventing cardiovascular disease, inflammatory diseases or even Alzheimer’s. On the other hand, the safety of fish and seafood is equally challenged and questions as these foods are very sensitive and highly perishable. On this down-side, they can account for food poisoning by the accumulation and formation of undesirable substances (e.g. biogenic amines, residual contaminants) or the out- break of various food-borne diseases due to microbial spoilage associated mainly with mismanagement in the cold chain and or other sources of contamination when hygienic precautions are handled with too much levity. The FishFermPlus ingredients and product applications will be assessed against potential contents of undesirable components and will impact on the overall safety by competing and interacting directly with the growth of the product-specific microbial flora and/or specific contaminating strains which constitute main microbial risks (e.g. Listeria sp.) in fish and seafood. Thereby the product safety as well as the individual shelf-life will be improved. In addition to bringing additional safety and higher convenience with longer storage times and higher flexibility in the consumption, it is envisaged to use the FishFermPlus project approach also for a second type of fermentation strategy. It will target to add to the taste and texture properties of fish or even other savoury foods since these sensory criteria are very crucial to consumer acceptance. Frequently they are also the reason why the consumption of fish foods is declined or only reluctantly followed to fulfil the minimum nutrition recommendations. With taste and texture enhanced products, light users of fish and seafood may increase their consumption to reach preferable nutrition goals rather than a set minimum, whereas heavy users will appreciate the additional variety of high-quality foods found on the market. All types of consumers will further benefit from a new natural processing and preservation method, which is also in line with currently fast-growing consumer demands.
Finally, the concept of the FishFermPlus project seizes its role in society and environmental aspects in keeping with sustainable development for the fish industry and reducing the burden of waste discarding. This is achieved firstly by making use of high-quality, under-utilised raw materials (e.g. from fillet production or other trimmings, from mere optical failure from mechanical damage/strain) for the fermentation process. Currently more than 30% of the harvested fish and seafood gets classified as a by-product and thrown away during the processing stages of the fish value chain, not taking into account food products that remain uneaten, but get thrown away in the package due to the indicated end of shelf-life, especially in supermarkets where legal requirements have to be fulfilled in terms of only distributing safe foods. The FishFermPlus results address this context from two angles: (1) It is made possible for European fish processors to act and produce more resource- efficient, which contributes significantly to saving the aquatic resources. (2) Less fish products will be discarded after production/sales, because of the prolonged shelf-life and improved logistics. Whilst raising such issues and the respective company profiles of SMEs that seek proactive solutions for a better resource-management and waste reduction, it can also be envisaged that the general consumer awareness will be finally attracted towards socially responsible fish production and consumption then which is the ultimate goal for a number of current EC policies in the area of fishery and fish production. In this respect, again, not only the social and environmental aspects have to be considered, but also the policy objectives of the Seventh Framework Programme with regard to SMEs und their role in contributing to the ERA. With little R&D expenditure in this specific field of the agro- food sector, FishFermPlus will assist the EC in raising substantially the innovation level in the fish industry in keeping with the Lisbon Strategy.
Essential for a successful market uptake of products and processes developed within the FishFermPlus project is a well-thought plan for the dissemination of project results and for the exploitation thereof. The aim is to advertise the special characteristics and advantages of the developed products and processes and to encourage the uptake into current customers’ product portfolio as well as to broaden the customer range. In this way, it is ensured that SMEs’ investment into the project will pay off and that the project results contribute to enhancing their competitiveness.
In addition to commercial benefits for the fish industry, the project also aims to extend the scientific state-of-the-art knowledge with regard to fermentation of fish raw material and related impact on sensory and safety characteristics. The knowledge can be used to develop manifold new products with added value beyond the aims and applications of the FishFermPlus project.
Therefore, a series of demonstration and dissemination activities has been conducted during the lifetime of the project and isintended for the post project period. The target group comprises actors of fish manufacturing industry, fish suppliers, and agencies for fish industry (resellers), food retailers, food wholesalers, convenience food producers, high class gastronomy and mass caterers, food associations, food-related magazines as well as the scientific community. In order to sufficiently reach the envisaged target group and spread project information effectively, a diverse and versatile strategy has to be followed. The relevant parties were addressed and will continuously be contacted via personal communication and consultation, preparation of flyers providing information about the project consortium and the activities in the project as well as product properties, project website with information about the consortium and project objectives as well as news and online surveys, articles in relevant magazines, press releases and announcements on partners’ websites, presentations and distribution of information at exhibitions, trade fairs, and congresses related to fish production, food-industry in general, and biotechnology, production of a video clip promoting the project results as well as demonstration events.
Exploitable foreground generated within the project covers five main aspects, namely the pre-competitive FishFermPlus fermentation process, the FishFermPlus starter cultures/fermentates, the FishFermPlus stabilization ingredients, the fermented and stabilized FishFermPlus intermediate product, and the FishFermPlus pilot product applications.
Project beneficiaries BG Salmon and Die Räucherei will invest in further optimization of process flow and economic efficiency as well as in further product application development. Developed products will be offered to different food sectors. Envisaged time to market is one year. Developed starter cultures (fermentates) and stabilization ingredients are ready for the market and will be exclusively delivered by project beneficiary Quality Food & Products to BG Salmon and Die Räucherei during one year after the project, in order to guarantee a competitive advantage until the products are launched on the market. Afterwards they will be freely available on the market. Project beneficiary ISI Food Protection will use the knowledge generated within the project for further strategic research including transfer of the process to other products and applications.
List of Websites:
Website address: www.fishfermplus.eu
Project Consortium:
ISI Food Protection (Coordinator) www.isifoodprotection.com
Dieter Elsser-Gravesen deg@isifoodprotection.com
Quality Food & Products www.quality-food-products.de
Jürgen Oltmer info@quality-food-products.de
Die Räucherei www.dieraeucherei.de
Hans-Joachim Kunkel j.kunkel@die-raeucherei.com
BG Production www.bgsalmon.dk
Gisle Olsen go@bgsalmon.dk
ttz Bremerhaven www.ttz-bremerhaven.de
Annika Gering agering@ttz-bremerhaven.de
Technical University of Denmark www.food.dtu.dk
Paw Dalgaard pada@food.dtu.dk