Final Report Summary - WHEYSAN (Natural whey-based disinfection formula: a solution for the sanitizing of whole and fresh-cut fruits and vegetables)
Executive Summary:
The market sales of fruits and vegetables (F&V) have grown rapidly in the recent decades as a result of the different changes in consumer attitudes. Extending the shelf-life of F&V products has become an important demand of the industry and consumers. Chlorine solutions have been widely used in the industry to retain the quality of F&V. However, the possible formation of carcinogenic chlorinated compounds in water has called into question the use of chlorine in food processing plants. Therefore, there is a real need to find new alternatives for the preservation of fruits and vegetables.
Whey is a by-product of the cheese industry. It contains a high level of minerals, bacteriocins and other low molecular bio-active peptides. Its high content of lactose makes it a high contaminant waste which needs to be processed before disposal. Big volumes of whey are generated and end up in rivers and seas without any type of treatment, causing terrible damage to the ecosystems of the affected areas. So, to reduce the high contaminant nature of whey it needs to be processed. Thus, being compliant with the European legislation regarding waste management.
Consequently, the aim of this project is focused in two major working lines: 1) To examine the influence of novel preservation methods based on whey, on the shelf-life of fruits and vegetables and 2) To process whey to achieve a profitable by-product with sanitizing properties.
For this purpose, a detailed literature review has been developed. The sanitizing of F&V for quality retention and microbial control has been studied, and the different analytical methods to evaluate the physicochemical, sensorial and microbiological activity and antioxidant parameters of the F&V have been described.
WHEYSAN presents new technologies for the preservation of whole and fresh-cut F&V, and for the processing of whey to achieve a profitable by-product. These technologies include the production of a sanitizing and antioxidant formula derived from whey, using the most appropriate treatments and the most appropriate efficient application techniques.
During the first nine months of work, RTD performers, DIT and CITA, have focused their activity in the production of the antioxidant and sanitizing formula out of whey derivatives. Several whey samples and derivatives from four different kinds of cheese whey have been analyzed in order to adapt the in-house methods to the analysis of the radical scavenging and antimicrobial activity.
During the first experiments, different types of whey were collected from the partner HFC. They were pasteurized using different combinations of time and temperature and a protocol of ultrafiltration and dialysis of whey was optimized. In total 16 different combinations of whey types and treatments were obtained and brought to the analytical lab for a further physico-chemical and microbial analysis.
The cheese whey had high antioxidant activity in terms of DPPH, FRAP, and ORAC; strong antimicrobial activity both in disk-diffusion method and broth-dilution. The whey treatment had positive impacts on the shelf life of a range of F&V tested. General acceptability, texture and colour were improved compared to control (water) and standard (chlorine) methods. Whey treatment also benefited the nutritional property of F&V applied at pilot and industrial scales.
Project Context and Objectives:
The purpose of the project WHEYSAN is to provide a solution for two major issues that currently affect the competitiveness of the participant SMEs: the management of whey and postharvest disinfection of fruits and vegetables (F&V). The whey is the residual liquid fraction that is obtained during the production of cheese. Its raised content of lactose makes it a high contaminant waste which needs to be processed before disposal. Whey and its derivatives have shown promising perspectives as natural preservatives to be used for disinfection of F&V as an alternative to chlorine, the most widely used disinfectant.
Postharvest disinfection of F&V is crucial for extending the market time of the produce. In recent years, a growing interest in the application of chlorine-free washing treatments for F&V based on natural preservatives has been observed because of the associated health and environmental risks. In particular, chlorine application is associated with the generation of carcinogenic halogenated disinfection by-products and the use of large amounts of water for the disinfection of F&V which suppose important economic and environmental implications. For these reasons, chlorine application is already forbidden in some European countries such as Germany, Netherlands, Denmark, Switzerland and Belgium.
Furthermore, the market sales of fruits and vegetables have grown rapidly in the recent decades as a result of the different changes in consumer attitudes. Extending the shelf-life of fruit and vegetable products has become an important demand of the industry and consumers.
WHEYSAN solution can provide SMEs in charge of commercializing the resultant sanitizer with an opportunity because of lower associated risks for human health and environment of the innovative formula.
The project WHEYSAN aims to develop application technologies for the processing of whey to achieve a profitable by-product, a new sanitizer, that improve the decontamination of whole and fresh-cut F&V and for the processing of whey to achieve a profitable by-product with sanitizing properties.
For this purpose, the whey produced by the dairy SME has been treated under different processing conditions and the efficacy of the whey derivatives as sanitizing agents for F&V have been assessed.
In order to make real the industrial exploitation of the new disinfection strategy, a commercial formula containing the whey derivatives was developed. For that, laboratory results have been scaled-up and the most efficient conditions for disinfection of whole and fresh-cut F&V have been defined at pilot and industrial scales.
The main objectives of the project are:
1. Production of the antioxidant and sanitizing formula out of whey derivatives.
a) Determining the proximate analysis of the different whey samples was studied in the project.
b) Adaptation of in-house methods for analyzing the radical scavenging activity of different whey samples and their derivatives.
c) Adaptation of in-house methods for analyzing the antimicrobial activity of different whey samples and their derivatives.
d) Adaption of in-house methods of analysis and quality evaluation of whole and fresh-cut fruits and vegetables.
e) Determination of the efficacy of whey samples and whey derivatives as sanitizing agent:
i. Determination of the antioxidant activity and relative antioxidant capacity index.
ii. Determination of antimicrobial activity.
f) Carry out the tests of functionality of whey samples with F&V at laboratory scale.
g) Characterization of composition and physicochemical properties of the successful whey samples.
h) Characterization of the whey samples with proven antioxidant and antimicrobial efficacy.
i. Determine the peptide profile of the whey and their derivatives.
ii. Isolation and characterization of antimicrobial compounds.
i) Optimization of the concentration of whey and whey derivatives to extend the shelf-life of whole and minimally processed fruits and vegetables.
j) Characterization of the most appropriate whey treatment giving rise to the most promising by-products to be used for decontamination of fruits and vegetables.
i. Development of the liquid matrix to be used for the delivery of sanitizing agents.
ii. Development of sanitizing formula prototypes for testing their efficacy at pilot and industrial scales.
2. Definition of the most efficient application techniques of the sanitizer at pilot and industrial scale.
a) Determination of the efficacy of whey sanitizing formula at pilot trial with whole and fresh-cut fruits and vegetables.
i. Testing of different application techniques of sanitizing in F&V.
ii. Assessing the quality during the shelf-life of fresh-cut F&V with whey application.
iii. Assessing the quality during the storage of whole F&V with whey application.
b) Compare the activity of whey with other sanitizing agents, overall, with chlorine.
c) Industrial trials of whey as sanitizing agent in whole and fresh-cut fruits and vegetables
i. Check the sanitizer stability over time according to the maximum production capacity.
ii. Evaluate the log reductions of microbial loads that naturally occur in raw F&V.
d) Organoleptic tests of the finished whole and fresh-cut F&V.
e) Implantation of a new wash system for fruit and vegetables, based on whey sanitizing agent.
With the aim of accomplishing these objectives, scientific characterization of WHEYSAN formula was done during the first period of the project. In the second one, antioxidant and antimicrobial activity of whey samples have been determined measured by different techniques (DPPH, FRAP, etc) and efficacy of whey samples as sanitizing agents for F&V has been studied. Moreover, whey samples have been applied to demonstrate project’s results at laboratory pilot and industrial scales. Additionally, project’s results have been disseminated by planning and undertaking different dissemination and training activities directed to both internal and external targets. Tasks relative to the project and consortium management have carried on during second period to ensure and accomplish project’s objectives.
Project Results:
During the first part of the project, various whey samples were supplied by partner HFC and processed (it can be seen in Figure 1 attached) to determine their anti-microbial activity and anti-oxidant capacity.
Moreover, the best laboratory protocols were selected for testing the efficacy of sanitizing products in fruits and vegetables, and they were used for the execution of further work packages. As a result, a Technical Sheet was developed for each product which describes the different analytical methods to evaluate the quality of each product. Subsequently, the most efficient methods reported in the literature for the application of sanitizing agents in different categories of fruits and vegetables at industrial scale were reviewed. Types of washers (immersion and non-immersion washers) and the factors influencing the efficacy of washing (time, sanitation practices, water quality, surfactants and antimicrobials and pathogens penetration) were reviewed. Also, the basics of fogging were explained, where Cold and Thermo Fogging are described.
In order to determine the microbial contribution of the whey samples to the F&V in which they were tested, the counts of mesophillic, psychotropic and lactic acid bacteria in whey samples were quantified by CITA. Then, the initial microbiological load of 18 whey batches, all of them coming from the production of the SME HFC, were analysed. Also, in order to reduce the initial load of the whey, two different pasteurization treatments were studied. Psychotrophic bacteria had the lower counts, whereas, the higher counts were observed in mesophillic aerobic bacteria, and in some cases, in moulds and yeasts.
The antimicrobial efficacy of whey and its derivatives was assessed in laboratory. The capacity was tested according to two different methods, which were developed during this task; disk-diffusion method and broth-dilution method. Alternaria was very sensible to most of the whey samples, although this effect was induced principally by the competition between Alternaria and a white mould (P. candida). Different results were obtained using the Disk-diffusion and Broth-dilution method, although these were complementary results. Both methods provide a clear antimicrobial effect which was different for each batch of whey analyzed.
To avoid a high Initial Microbial Load of Whey samples, transport conditions were improved and heat treatment process was carried out in the in CITA and DIT facilities.
Then, the selection of the best batch of whey was performed based on the best results of antimicrobial activity (both in disk-diffusion method and broth-dilution method), antioxidant activity (in terms of DPPH, FRAP and ORAC), and had the lowest initial microbial load.
The next step was optimizing the concentration of whey samples to extend the shelf-life of F&V and comparing the activity of whey with chlorine. Physio-chemical analysis and microbiological properties have been monitored and specific pathogen and spoilage microorganisms have also been investigated. The whey treatment had positive impacts on the shelf life of a range of whole and minimally processed F&V tested. General acceptability, texture and color were improved compared to control (water) and standard (chlorine) methods.
Regarding the antimicrobial effect of whey, only the treatment by Dipping in chlorinated water bath with 150 ppm NaOCl was better results than the treatment by Dipping in Whey. Results of the treatments by Dipping in water bath with 50 ppm NaOCl, bath with 100 ppm NaOCl and Dipping in Whey were similar.
After that, a review of possible adjuvants that enhance the thermal fogging of aqueous solutions have been carried out as well as the testing of different application techniques at pilot scale was performed. The equipment used for Cold-fogging was AEROTURBEX which was a very fine atomization without high temperature. The results were excellent after treatment indoor use. The equipment used for Thermo-fogging was AEROBRUMER for working at low temperature (100 0C) and DIESEL machine for working at high temperature (260-3000C).The main difference between them was the drop size of the fog (AEROBRUMER generates higher drop size than DIESEL machine). (Figure 2, 3&4 attached).
Several samples of whole and fresh-cut F&V were treated with the best formula prototype and with the best application method. Afterwards, the changes on physicochemical and microbial counts were assessed during the storage time of the whole and fresh-cut F&V.
AGF carried out the different application techniques using their equipment and their Know how on the facilities of CITA which carried out the analytical studies to assess the influence of whey in their shelf-life. For whole products, on day 0, most of the treatments tested microbial reductions were obtained. In the best case, the chlorine treatment achieves a reduction of 1 logarithmic unit. However, the counts are equalized with increasing storage time. For Fresh-cut products inoculated with E. coli and Salmonella, all treatments were effective and able to reduce E. coli and Salmonella counts over the shelf-life of the products.
For testing the sanitizing efficacy of whey at industrial scale, the three whey-based formulas developed were tested on different representative types of whole F&V: apple, onion, carrot, tomato, lettuce and cabbage (Figure 5&6 attached). For fresh-cut F&V were used: lettuce, broccoli, carrot and apple.
The results showed for most of whey products no undesirable aroma detected during the shelf-life of the product in any of the treatments. For General Acceptability, whey by cold-fogging was the best treatment for fresh-cut broccoli and carrot, whey by dipping was the best treatment for fresh-cut apple and control treatment for fresh-cut lettuce. The Whey by cold-fogging treatment has had a highly positive effect on the cut color of fresh-cut broccoli.
The main expected final results for the whole project are included below. They have been successfully achieved.
• Result 1: Development of whey-based raw material with sanitizing properties.
Whey is a by-product of Cheese production and is waste for the SME dairies and cheese processors. It is costly to dispose of and due to high BOD is normally sent to farmers for animal feed. This Project addresses both the environmental impact of this waste source and the development of a natural sanitizer. Cheese whey has good antioxidant activity so when applied to food products preserves shelf-life and promotes its nutritional value. Moreover, cheese whey has good antimicrobial activity which makes it suitable as a healthy natural alternative preservative. Furthermore, cheese whey treatment has a positive impact on the shelf life and nutritional value of Fruit and vegetables (both whole and minimally processed).
The best batch of whey has been selected. It got the best results of antimicrobial activity (both in disk-diffusion method and broth-dilution method), antioxidant activity (in terms of DPPH, FRAP and ORAC), and had the lowest initial microbial load.
• Result 2: Development of whey based commercial formula.
It represents the main breakthrough of the project. Full characteristics of whey formula have been defined in order to undertake commercial application.
The final product formulation was produced through the addition of the whey-based sanitizing formula in to a liquid matrix specifically suited for commercial application. Three different prototypes were developed and their efficacy was tested in whole and fresh-cut F&V at pilot and industrial scales.
• Result 3: Definition of the industrial protocol for whey processing.
At the end of the project a protocol for whey processing has been reported. HFC could implement in its facilities the process for whey treatment or HFC could license these rights in order to being exploited by other companies that actually treat whey for other applications.
In addition to development of the most efficient whey processing protocol, it is readily scaled up to Industrial scale after the testing of different application techniques at pilot scale.
• Result 4: Industrial application of the whey-based formula for disinfection of whole F&V.
Several samples of whole F&V have been treated with the best formula prototype and with the best application method. Industrial applications have been carried out by partners CITA and AGF at CATEC’s facilities. Used techniques are: Bath; Cold-fogging and Thermo-fogging.
• Result 5: Industrial application of the whey-based formula for disinfection of fresh-cuts F&V.
Several samples of fresh-cuts F&V have been treated with the best formula prototype and with the best application method. Industrial applications have been carried out by partners CITA and AGF at CITA’s facilities with fruits delivered by CITRUS.
Potential Impact:
The main result of the project WHEYSAN is to present new technologies for the decontamination of whole and fresh-cut fruits and vegetables (F&V), as well as for the processing of whey to achieve a profitable by-product with sanitizing properties for the disinfection processes.
Thus, the main innovation of the project WHEYSAN is based on the development of a sanitizing and antioxidant formula out of whey, which will allow a more efficient and sustainable decontamination of F&V than the current synthetic and chemical-based washing treatments.
Whey and its derivatives have shown promising perspectives as natural preservatives to be used for disinfection of F&V as an alternative to current ones used. The sanitizing agents used up to now are chemical-based: chlorine, chlorine dioxide, organic acids, hydrogen peroxide, calcium-based solutions, ozone and electrolyzed water. Using whey derivatives appears to be less polluting and more effective.
In recent years, a growing interest in the application of chlorine-free washing treatments for F&V based on natural preservatives has been observed. Chlorine is the most effective sanitizer among the currently used ones by the F&V industry to ensure food safety. However, there is a trend towards eliminating chlorine from disinfection processes because of the associated health and environmental risks. In particular, it is associated with the generation of carcinogenic halogenated disinfection by-products and the use of large amounts of water for the disinfection of F&V, which suppose important economic and environmental implications. WHEYSAN solution can provide SMEs in charge of commercializing the resultant sanitizer with an opportunity because of lower associated risks for human and environment health of the innovative formula. It also provides a solution to the F&V processors and handlers. Moreover, the use of chlorine for the disinfection of F&V is forbidden in some European countries: Germany, Netherlands, Denmark, Switzerland and Belgium. Thus, new alternatives that are healthier for humans, friendlier with the environment and safer for consumers are required. In some of these countries, the use of big quantities of water is recommended. Nevertheless, the risk of cross-contamination is not removed without adding a sanitizer.
In this context, the main purpose of the project WHEYSAN is to develop a natural and whey-based sanitizer that is suitable for the disinfection process of the SME participants in the industry of whole and minimally-processed F&V. In the meantime, the project provides a solution to the dairy SMEs, which gain the know-how for developing a new value-added whey by-product with sanitizing properties.
The results of WHEYSAN have significant impact on different markets. They are related to the markets where the innovative generated products compete. Main target markets for the products resulting from the WHEYSAN project are: on the one hand, the market of food preservatives and more specifically, the segment of sanitizer for decontamination and disinfection of whole and fresh-cut F&V, and on the other and, the manufacturing sector of dairy products.
In the market of sanitizer for decontamination and disinfection of whole and fresh-cut F&V we can distinguish two target groups: 1) Companies that currently commercialize disinfectants and other products for the cleaning and sanitizing, which could be the intermediaries for the commercialization of the formula (e.g. CITROSOL, TECNIDEX, FOMESA…), and 2) F&V growers and F&V cooperatives, which are the users: cleaning their fruits with the formula obtained from WHEYSAN. In this market, the segment of organic F&V has been identified as relevant due to the sustainable character of the innovative formula.
In the market of dairy products, the target users are companies focused on cheese production because it is where whey would be obtained from in order to develop a new value-added whey by-product with sanitizing properties.
The four SMEs members of the WHEYSAN Consortium are solid and well established companies in their local markets and even in different geographical areas. They work in the target markets of the project. Through this project they are going to be able to access a network of contacts within their own market sector. They will implement project´s results, in terms of cost cuts, enhanced competitive benefits and advantages in the global markets. In general terms, every SME participant in the project depend on each other for the successful exploitation of the project’s results. HFC, ECO and AGF cannot develop the commercial formula on their own; they need cooperation with other partners. At the same time, CATEC will not be able to apply the formula on its products if others do not develop it. HFC will gain a profit from whey and will diversify its businesses through commercialization of whey by-products with sanitizing properties.
AGROFIELD (AGF)
This company is focused on the cleaning/disinfection of agro alimentary products. From the WHEYSAN project they expect to gain access to the exploitation rights of a novel and natural based sanitizing agent with proven efficacy as a disinfectant for F&V and a functional alternative for the replacement of chlorine.
AGF will create a new source of revenue following the commercial application of the new whey-based formula through their current systems for industrial disinfection of F&V.
They will install adequate machinery for the implementation of the product, perform maintenance tasks, make the different applications on the customer´s premises and they will be in charge of the product sales and distribution, mainly in their national market (Spain) and Portugal, where they currently have important clients and contacts.
HIGHLAND FINE CHEESES (HFC)
They are artisan cheese producers of traditional fresh lactic cheese, mould ripened and cheddar. In order to be compliant with the European legislation they need to find a solution for the whey resulting from their production process, therefore they want to produce a value added by-product
They will capture whey and bring to market a value added product to reduce their total waste. Thus, they will gain a profit from whey and will diversify its business through the commercialization of whey by-products with sanitizing properties.
CAMPOTEC Ltd. (CATEC)
CATEC is engaged in the sale of fruits (pomoideae and prunoideae), potatoes, vegetables and minimally-processed horticultural products. Its main goal is the production of high quality agricultural products with the latest technology. They have validated the sanitizing activity of the WHEY based commercial formula within its industrial facilities, for the washing and disinfection of whole and fresh-cut F&V produce. They want to gain access to the exploitation rights of the know-how regarding the best way to apply the whey based sanitizing formula within their disinfection operations.
They will also benefit from an increased market time of their F&V, and from a better consumer acceptance of their produce, treated with natural disinfectants alternative to chlorine.
ECOSOIL (ECO)
In their daily activity they are focused on getting liquid formulas used as natural fertilizers that can provide quality materials and minerals to the organic horticultural products growth. During the project execution, they have developed the liquid sanitizing formula derived from whey. Their aim is to gain access to the exploitation rights of the whey based sanitizing formula and to enter in the EU market of food preservatives. In their activities, they will take into account Health and Environmental policies. Thus, ECO will increase its pipeline through the commercialization of a natural product that is suitable for the disinfection of whole and fresh-cut F&V.
The SME participants, in their search for effective new sainting agents, often operate within areas of regulatory uncertainty, with a fairly limited access to technology as well as a low technical expertise and research capacity. Working in a multidisciplinary team, these problems are solved. Thus, the support of experienced RTDs is essential to acquire the technological know-how for the development of an innovative and sustainable sanitizer.
List of Websites:
PUBLIC WEBSITE ADDRESS: http://www.contactica.es
CONTACT DETAILS:
Project Coordinator: Agrofield
Person in charge: Rodrigo Casas Milagro
Telephone: +34 94 145 20 33
Fax: + 34 94 144 88 92
e-mail: agrofield@agrofield.es
The market sales of fruits and vegetables (F&V) have grown rapidly in the recent decades as a result of the different changes in consumer attitudes. Extending the shelf-life of F&V products has become an important demand of the industry and consumers. Chlorine solutions have been widely used in the industry to retain the quality of F&V. However, the possible formation of carcinogenic chlorinated compounds in water has called into question the use of chlorine in food processing plants. Therefore, there is a real need to find new alternatives for the preservation of fruits and vegetables.
Whey is a by-product of the cheese industry. It contains a high level of minerals, bacteriocins and other low molecular bio-active peptides. Its high content of lactose makes it a high contaminant waste which needs to be processed before disposal. Big volumes of whey are generated and end up in rivers and seas without any type of treatment, causing terrible damage to the ecosystems of the affected areas. So, to reduce the high contaminant nature of whey it needs to be processed. Thus, being compliant with the European legislation regarding waste management.
Consequently, the aim of this project is focused in two major working lines: 1) To examine the influence of novel preservation methods based on whey, on the shelf-life of fruits and vegetables and 2) To process whey to achieve a profitable by-product with sanitizing properties.
For this purpose, a detailed literature review has been developed. The sanitizing of F&V for quality retention and microbial control has been studied, and the different analytical methods to evaluate the physicochemical, sensorial and microbiological activity and antioxidant parameters of the F&V have been described.
WHEYSAN presents new technologies for the preservation of whole and fresh-cut F&V, and for the processing of whey to achieve a profitable by-product. These technologies include the production of a sanitizing and antioxidant formula derived from whey, using the most appropriate treatments and the most appropriate efficient application techniques.
During the first nine months of work, RTD performers, DIT and CITA, have focused their activity in the production of the antioxidant and sanitizing formula out of whey derivatives. Several whey samples and derivatives from four different kinds of cheese whey have been analyzed in order to adapt the in-house methods to the analysis of the radical scavenging and antimicrobial activity.
During the first experiments, different types of whey were collected from the partner HFC. They were pasteurized using different combinations of time and temperature and a protocol of ultrafiltration and dialysis of whey was optimized. In total 16 different combinations of whey types and treatments were obtained and brought to the analytical lab for a further physico-chemical and microbial analysis.
The cheese whey had high antioxidant activity in terms of DPPH, FRAP, and ORAC; strong antimicrobial activity both in disk-diffusion method and broth-dilution. The whey treatment had positive impacts on the shelf life of a range of F&V tested. General acceptability, texture and colour were improved compared to control (water) and standard (chlorine) methods. Whey treatment also benefited the nutritional property of F&V applied at pilot and industrial scales.
Project Context and Objectives:
The purpose of the project WHEYSAN is to provide a solution for two major issues that currently affect the competitiveness of the participant SMEs: the management of whey and postharvest disinfection of fruits and vegetables (F&V). The whey is the residual liquid fraction that is obtained during the production of cheese. Its raised content of lactose makes it a high contaminant waste which needs to be processed before disposal. Whey and its derivatives have shown promising perspectives as natural preservatives to be used for disinfection of F&V as an alternative to chlorine, the most widely used disinfectant.
Postharvest disinfection of F&V is crucial for extending the market time of the produce. In recent years, a growing interest in the application of chlorine-free washing treatments for F&V based on natural preservatives has been observed because of the associated health and environmental risks. In particular, chlorine application is associated with the generation of carcinogenic halogenated disinfection by-products and the use of large amounts of water for the disinfection of F&V which suppose important economic and environmental implications. For these reasons, chlorine application is already forbidden in some European countries such as Germany, Netherlands, Denmark, Switzerland and Belgium.
Furthermore, the market sales of fruits and vegetables have grown rapidly in the recent decades as a result of the different changes in consumer attitudes. Extending the shelf-life of fruit and vegetable products has become an important demand of the industry and consumers.
WHEYSAN solution can provide SMEs in charge of commercializing the resultant sanitizer with an opportunity because of lower associated risks for human health and environment of the innovative formula.
The project WHEYSAN aims to develop application technologies for the processing of whey to achieve a profitable by-product, a new sanitizer, that improve the decontamination of whole and fresh-cut F&V and for the processing of whey to achieve a profitable by-product with sanitizing properties.
For this purpose, the whey produced by the dairy SME has been treated under different processing conditions and the efficacy of the whey derivatives as sanitizing agents for F&V have been assessed.
In order to make real the industrial exploitation of the new disinfection strategy, a commercial formula containing the whey derivatives was developed. For that, laboratory results have been scaled-up and the most efficient conditions for disinfection of whole and fresh-cut F&V have been defined at pilot and industrial scales.
The main objectives of the project are:
1. Production of the antioxidant and sanitizing formula out of whey derivatives.
a) Determining the proximate analysis of the different whey samples was studied in the project.
b) Adaptation of in-house methods for analyzing the radical scavenging activity of different whey samples and their derivatives.
c) Adaptation of in-house methods for analyzing the antimicrobial activity of different whey samples and their derivatives.
d) Adaption of in-house methods of analysis and quality evaluation of whole and fresh-cut fruits and vegetables.
e) Determination of the efficacy of whey samples and whey derivatives as sanitizing agent:
i. Determination of the antioxidant activity and relative antioxidant capacity index.
ii. Determination of antimicrobial activity.
f) Carry out the tests of functionality of whey samples with F&V at laboratory scale.
g) Characterization of composition and physicochemical properties of the successful whey samples.
h) Characterization of the whey samples with proven antioxidant and antimicrobial efficacy.
i. Determine the peptide profile of the whey and their derivatives.
ii. Isolation and characterization of antimicrobial compounds.
i) Optimization of the concentration of whey and whey derivatives to extend the shelf-life of whole and minimally processed fruits and vegetables.
j) Characterization of the most appropriate whey treatment giving rise to the most promising by-products to be used for decontamination of fruits and vegetables.
i. Development of the liquid matrix to be used for the delivery of sanitizing agents.
ii. Development of sanitizing formula prototypes for testing their efficacy at pilot and industrial scales.
2. Definition of the most efficient application techniques of the sanitizer at pilot and industrial scale.
a) Determination of the efficacy of whey sanitizing formula at pilot trial with whole and fresh-cut fruits and vegetables.
i. Testing of different application techniques of sanitizing in F&V.
ii. Assessing the quality during the shelf-life of fresh-cut F&V with whey application.
iii. Assessing the quality during the storage of whole F&V with whey application.
b) Compare the activity of whey with other sanitizing agents, overall, with chlorine.
c) Industrial trials of whey as sanitizing agent in whole and fresh-cut fruits and vegetables
i. Check the sanitizer stability over time according to the maximum production capacity.
ii. Evaluate the log reductions of microbial loads that naturally occur in raw F&V.
d) Organoleptic tests of the finished whole and fresh-cut F&V.
e) Implantation of a new wash system for fruit and vegetables, based on whey sanitizing agent.
With the aim of accomplishing these objectives, scientific characterization of WHEYSAN formula was done during the first period of the project. In the second one, antioxidant and antimicrobial activity of whey samples have been determined measured by different techniques (DPPH, FRAP, etc) and efficacy of whey samples as sanitizing agents for F&V has been studied. Moreover, whey samples have been applied to demonstrate project’s results at laboratory pilot and industrial scales. Additionally, project’s results have been disseminated by planning and undertaking different dissemination and training activities directed to both internal and external targets. Tasks relative to the project and consortium management have carried on during second period to ensure and accomplish project’s objectives.
Project Results:
During the first part of the project, various whey samples were supplied by partner HFC and processed (it can be seen in Figure 1 attached) to determine their anti-microbial activity and anti-oxidant capacity.
Moreover, the best laboratory protocols were selected for testing the efficacy of sanitizing products in fruits and vegetables, and they were used for the execution of further work packages. As a result, a Technical Sheet was developed for each product which describes the different analytical methods to evaluate the quality of each product. Subsequently, the most efficient methods reported in the literature for the application of sanitizing agents in different categories of fruits and vegetables at industrial scale were reviewed. Types of washers (immersion and non-immersion washers) and the factors influencing the efficacy of washing (time, sanitation practices, water quality, surfactants and antimicrobials and pathogens penetration) were reviewed. Also, the basics of fogging were explained, where Cold and Thermo Fogging are described.
In order to determine the microbial contribution of the whey samples to the F&V in which they were tested, the counts of mesophillic, psychotropic and lactic acid bacteria in whey samples were quantified by CITA. Then, the initial microbiological load of 18 whey batches, all of them coming from the production of the SME HFC, were analysed. Also, in order to reduce the initial load of the whey, two different pasteurization treatments were studied. Psychotrophic bacteria had the lower counts, whereas, the higher counts were observed in mesophillic aerobic bacteria, and in some cases, in moulds and yeasts.
The antimicrobial efficacy of whey and its derivatives was assessed in laboratory. The capacity was tested according to two different methods, which were developed during this task; disk-diffusion method and broth-dilution method. Alternaria was very sensible to most of the whey samples, although this effect was induced principally by the competition between Alternaria and a white mould (P. candida). Different results were obtained using the Disk-diffusion and Broth-dilution method, although these were complementary results. Both methods provide a clear antimicrobial effect which was different for each batch of whey analyzed.
To avoid a high Initial Microbial Load of Whey samples, transport conditions were improved and heat treatment process was carried out in the in CITA and DIT facilities.
Then, the selection of the best batch of whey was performed based on the best results of antimicrobial activity (both in disk-diffusion method and broth-dilution method), antioxidant activity (in terms of DPPH, FRAP and ORAC), and had the lowest initial microbial load.
The next step was optimizing the concentration of whey samples to extend the shelf-life of F&V and comparing the activity of whey with chlorine. Physio-chemical analysis and microbiological properties have been monitored and specific pathogen and spoilage microorganisms have also been investigated. The whey treatment had positive impacts on the shelf life of a range of whole and minimally processed F&V tested. General acceptability, texture and color were improved compared to control (water) and standard (chlorine) methods.
Regarding the antimicrobial effect of whey, only the treatment by Dipping in chlorinated water bath with 150 ppm NaOCl was better results than the treatment by Dipping in Whey. Results of the treatments by Dipping in water bath with 50 ppm NaOCl, bath with 100 ppm NaOCl and Dipping in Whey were similar.
After that, a review of possible adjuvants that enhance the thermal fogging of aqueous solutions have been carried out as well as the testing of different application techniques at pilot scale was performed. The equipment used for Cold-fogging was AEROTURBEX which was a very fine atomization without high temperature. The results were excellent after treatment indoor use. The equipment used for Thermo-fogging was AEROBRUMER for working at low temperature (100 0C) and DIESEL machine for working at high temperature (260-3000C).The main difference between them was the drop size of the fog (AEROBRUMER generates higher drop size than DIESEL machine). (Figure 2, 3&4 attached).
Several samples of whole and fresh-cut F&V were treated with the best formula prototype and with the best application method. Afterwards, the changes on physicochemical and microbial counts were assessed during the storage time of the whole and fresh-cut F&V.
AGF carried out the different application techniques using their equipment and their Know how on the facilities of CITA which carried out the analytical studies to assess the influence of whey in their shelf-life. For whole products, on day 0, most of the treatments tested microbial reductions were obtained. In the best case, the chlorine treatment achieves a reduction of 1 logarithmic unit. However, the counts are equalized with increasing storage time. For Fresh-cut products inoculated with E. coli and Salmonella, all treatments were effective and able to reduce E. coli and Salmonella counts over the shelf-life of the products.
For testing the sanitizing efficacy of whey at industrial scale, the three whey-based formulas developed were tested on different representative types of whole F&V: apple, onion, carrot, tomato, lettuce and cabbage (Figure 5&6 attached). For fresh-cut F&V were used: lettuce, broccoli, carrot and apple.
The results showed for most of whey products no undesirable aroma detected during the shelf-life of the product in any of the treatments. For General Acceptability, whey by cold-fogging was the best treatment for fresh-cut broccoli and carrot, whey by dipping was the best treatment for fresh-cut apple and control treatment for fresh-cut lettuce. The Whey by cold-fogging treatment has had a highly positive effect on the cut color of fresh-cut broccoli.
The main expected final results for the whole project are included below. They have been successfully achieved.
• Result 1: Development of whey-based raw material with sanitizing properties.
Whey is a by-product of Cheese production and is waste for the SME dairies and cheese processors. It is costly to dispose of and due to high BOD is normally sent to farmers for animal feed. This Project addresses both the environmental impact of this waste source and the development of a natural sanitizer. Cheese whey has good antioxidant activity so when applied to food products preserves shelf-life and promotes its nutritional value. Moreover, cheese whey has good antimicrobial activity which makes it suitable as a healthy natural alternative preservative. Furthermore, cheese whey treatment has a positive impact on the shelf life and nutritional value of Fruit and vegetables (both whole and minimally processed).
The best batch of whey has been selected. It got the best results of antimicrobial activity (both in disk-diffusion method and broth-dilution method), antioxidant activity (in terms of DPPH, FRAP and ORAC), and had the lowest initial microbial load.
• Result 2: Development of whey based commercial formula.
It represents the main breakthrough of the project. Full characteristics of whey formula have been defined in order to undertake commercial application.
The final product formulation was produced through the addition of the whey-based sanitizing formula in to a liquid matrix specifically suited for commercial application. Three different prototypes were developed and their efficacy was tested in whole and fresh-cut F&V at pilot and industrial scales.
• Result 3: Definition of the industrial protocol for whey processing.
At the end of the project a protocol for whey processing has been reported. HFC could implement in its facilities the process for whey treatment or HFC could license these rights in order to being exploited by other companies that actually treat whey for other applications.
In addition to development of the most efficient whey processing protocol, it is readily scaled up to Industrial scale after the testing of different application techniques at pilot scale.
• Result 4: Industrial application of the whey-based formula for disinfection of whole F&V.
Several samples of whole F&V have been treated with the best formula prototype and with the best application method. Industrial applications have been carried out by partners CITA and AGF at CATEC’s facilities. Used techniques are: Bath; Cold-fogging and Thermo-fogging.
• Result 5: Industrial application of the whey-based formula for disinfection of fresh-cuts F&V.
Several samples of fresh-cuts F&V have been treated with the best formula prototype and with the best application method. Industrial applications have been carried out by partners CITA and AGF at CITA’s facilities with fruits delivered by CITRUS.
Potential Impact:
The main result of the project WHEYSAN is to present new technologies for the decontamination of whole and fresh-cut fruits and vegetables (F&V), as well as for the processing of whey to achieve a profitable by-product with sanitizing properties for the disinfection processes.
Thus, the main innovation of the project WHEYSAN is based on the development of a sanitizing and antioxidant formula out of whey, which will allow a more efficient and sustainable decontamination of F&V than the current synthetic and chemical-based washing treatments.
Whey and its derivatives have shown promising perspectives as natural preservatives to be used for disinfection of F&V as an alternative to current ones used. The sanitizing agents used up to now are chemical-based: chlorine, chlorine dioxide, organic acids, hydrogen peroxide, calcium-based solutions, ozone and electrolyzed water. Using whey derivatives appears to be less polluting and more effective.
In recent years, a growing interest in the application of chlorine-free washing treatments for F&V based on natural preservatives has been observed. Chlorine is the most effective sanitizer among the currently used ones by the F&V industry to ensure food safety. However, there is a trend towards eliminating chlorine from disinfection processes because of the associated health and environmental risks. In particular, it is associated with the generation of carcinogenic halogenated disinfection by-products and the use of large amounts of water for the disinfection of F&V, which suppose important economic and environmental implications. WHEYSAN solution can provide SMEs in charge of commercializing the resultant sanitizer with an opportunity because of lower associated risks for human and environment health of the innovative formula. It also provides a solution to the F&V processors and handlers. Moreover, the use of chlorine for the disinfection of F&V is forbidden in some European countries: Germany, Netherlands, Denmark, Switzerland and Belgium. Thus, new alternatives that are healthier for humans, friendlier with the environment and safer for consumers are required. In some of these countries, the use of big quantities of water is recommended. Nevertheless, the risk of cross-contamination is not removed without adding a sanitizer.
In this context, the main purpose of the project WHEYSAN is to develop a natural and whey-based sanitizer that is suitable for the disinfection process of the SME participants in the industry of whole and minimally-processed F&V. In the meantime, the project provides a solution to the dairy SMEs, which gain the know-how for developing a new value-added whey by-product with sanitizing properties.
The results of WHEYSAN have significant impact on different markets. They are related to the markets where the innovative generated products compete. Main target markets for the products resulting from the WHEYSAN project are: on the one hand, the market of food preservatives and more specifically, the segment of sanitizer for decontamination and disinfection of whole and fresh-cut F&V, and on the other and, the manufacturing sector of dairy products.
In the market of sanitizer for decontamination and disinfection of whole and fresh-cut F&V we can distinguish two target groups: 1) Companies that currently commercialize disinfectants and other products for the cleaning and sanitizing, which could be the intermediaries for the commercialization of the formula (e.g. CITROSOL, TECNIDEX, FOMESA…), and 2) F&V growers and F&V cooperatives, which are the users: cleaning their fruits with the formula obtained from WHEYSAN. In this market, the segment of organic F&V has been identified as relevant due to the sustainable character of the innovative formula.
In the market of dairy products, the target users are companies focused on cheese production because it is where whey would be obtained from in order to develop a new value-added whey by-product with sanitizing properties.
The four SMEs members of the WHEYSAN Consortium are solid and well established companies in their local markets and even in different geographical areas. They work in the target markets of the project. Through this project they are going to be able to access a network of contacts within their own market sector. They will implement project´s results, in terms of cost cuts, enhanced competitive benefits and advantages in the global markets. In general terms, every SME participant in the project depend on each other for the successful exploitation of the project’s results. HFC, ECO and AGF cannot develop the commercial formula on their own; they need cooperation with other partners. At the same time, CATEC will not be able to apply the formula on its products if others do not develop it. HFC will gain a profit from whey and will diversify its businesses through commercialization of whey by-products with sanitizing properties.
AGROFIELD (AGF)
This company is focused on the cleaning/disinfection of agro alimentary products. From the WHEYSAN project they expect to gain access to the exploitation rights of a novel and natural based sanitizing agent with proven efficacy as a disinfectant for F&V and a functional alternative for the replacement of chlorine.
AGF will create a new source of revenue following the commercial application of the new whey-based formula through their current systems for industrial disinfection of F&V.
They will install adequate machinery for the implementation of the product, perform maintenance tasks, make the different applications on the customer´s premises and they will be in charge of the product sales and distribution, mainly in their national market (Spain) and Portugal, where they currently have important clients and contacts.
HIGHLAND FINE CHEESES (HFC)
They are artisan cheese producers of traditional fresh lactic cheese, mould ripened and cheddar. In order to be compliant with the European legislation they need to find a solution for the whey resulting from their production process, therefore they want to produce a value added by-product
They will capture whey and bring to market a value added product to reduce their total waste. Thus, they will gain a profit from whey and will diversify its business through the commercialization of whey by-products with sanitizing properties.
CAMPOTEC Ltd. (CATEC)
CATEC is engaged in the sale of fruits (pomoideae and prunoideae), potatoes, vegetables and minimally-processed horticultural products. Its main goal is the production of high quality agricultural products with the latest technology. They have validated the sanitizing activity of the WHEY based commercial formula within its industrial facilities, for the washing and disinfection of whole and fresh-cut F&V produce. They want to gain access to the exploitation rights of the know-how regarding the best way to apply the whey based sanitizing formula within their disinfection operations.
They will also benefit from an increased market time of their F&V, and from a better consumer acceptance of their produce, treated with natural disinfectants alternative to chlorine.
ECOSOIL (ECO)
In their daily activity they are focused on getting liquid formulas used as natural fertilizers that can provide quality materials and minerals to the organic horticultural products growth. During the project execution, they have developed the liquid sanitizing formula derived from whey. Their aim is to gain access to the exploitation rights of the whey based sanitizing formula and to enter in the EU market of food preservatives. In their activities, they will take into account Health and Environmental policies. Thus, ECO will increase its pipeline through the commercialization of a natural product that is suitable for the disinfection of whole and fresh-cut F&V.
The SME participants, in their search for effective new sainting agents, often operate within areas of regulatory uncertainty, with a fairly limited access to technology as well as a low technical expertise and research capacity. Working in a multidisciplinary team, these problems are solved. Thus, the support of experienced RTDs is essential to acquire the technological know-how for the development of an innovative and sustainable sanitizer.
List of Websites:
PUBLIC WEBSITE ADDRESS: http://www.contactica.es
CONTACT DETAILS:
Project Coordinator: Agrofield
Person in charge: Rodrigo Casas Milagro
Telephone: +34 94 145 20 33
Fax: + 34 94 144 88 92
e-mail: agrofield@agrofield.es
