bere caso cerveceros espana

This case study has been produced for Fundación Innova by Camila Lee No, under the supervision of Adrián Escardino Malva. Fundación Innova is the management body of Valencia Polytechnic University‘s Science Park: La Ciudad Politécnica de la Innovación, and it produces case studies to show the dynamics of cooperation between the researchers at the Park and their business environment. Fundación Innova would like to thank the Cerveceros de España association and the researcher of the Immunotechnology Group at Valencia Polytechnic University, Dr Ángel Montoya Baides for their collaboration.

Copyright 2005 Fundación Innova. Reproduction of this document is authorised for information and non-profit purposes only.

Case Studies by Fundación Innova: development of an immunodetection system for microbiological control of beer. of 8

DEVELOPMENT OF AN IMMUNODETECTION SYSTEM FOR MICROBIOLOGICAL CONTROL OF BEER

“Generically speaking, Innovation and Development in the Food and Agriculture Industry revolve around two fundamental aspects: the safety and quality of foodstuffs.” “In both cases the main concern is to develop molecular methods of detection, analysis and diagnosis that are fast, highly sensitive and that enable computerised tracking of a wide range of agents that could be harmful in food products.”

Applications of biosensors in the food and agriculture industry Technological vigilance report; Madrid R&D.

The development of new highly specific molecular analysis systems with high sensitivity and low response times is particularly important in the medical industry. However, it has recently taken on increasing importance in environmental areas and is gradually becoming more widespread in food and agriculture companies.

In the coming years, it will be normal to see their use in automatic quality checking systems, health inspections and process control systems in the food and agriculture sector.

This case study deals with the collaboration between the Spanish brewery association Cerveceros de España and Valencia Polytechnic University to develop an analysis system for microbiological control of beer based on the use of biosensors and monoclonal antibodies.

ORIGIN OF THE PROJECT: CERVECEROS DE ESPAÑA Cerveceros de España (http://www.cerveceros.org) is an association that was founded in 1995 to represent Spain’s most important breweries. It originally goes back to 1922, when the “Association of Spanish Breweries” was founded. Its current members are Heineken España, Mahou-San Miguel, Damm, Compañía Cervecera de Canarias, Hijos de Rivera, and La Zaragozana, which account for almost all Spain’s beer production.

Cerveceros de España is an active member of the European Brewers Association and the Confederation of EU Food and Drink Industries, and it is also involved in the European Brewery Convention, the organisation responsible for researching and improving the quality of beer.

THE PROBLEM / OPPORTUNITY IN THE BEER INDUSTRY

Food spoilage due to the action of undesired microorganisms is one of the main reasons for loss of quality and health hazards in the food and agriculture industry. In the case of beer production, lactic acid bacteria (specifically those of the genera Lactobacillus and Pediococcus) are most often responsible for this problem. The effects of this may be noticed as cloudiness, strange flavours, an increased concentration of biogenic amines, increased viscosity, increased acidity, etc.

Fermentation vats at the Cervezas Cruzcampo brewery(Grupo Heineken)


Traditional microbiological control methods in the beer industry are based on the isolation and concentration of microorganisms in culture media until a sufficiently large population is achieved for them to be identified. The main drawbacks of this are its low specificity and the long incubation periods required to separate and concentrate the bacteria before they reach detectable levels (normally 5 to 8 days). The time need to ensure the level of quality and safety of each batch means that companies have to keep a considerable amount of the finished product in stock, with the consequent impact on the structure of their circulating capital.

“This is particularly important during the summer months, the time of year when our members have to keep very large amounts of the product in stock in order to meet the high demand of the market. In a country where the average daily consumption is approximately 12 million litres in summer, keeping six days’ production in stock has an impact of over 80 million euros on the sector’s circulating capital. Our members were therefore clearly interested in reducing this ratio and, at the same time, improving the reliability of the controls.”

Jacobo Olaya (General Manager of Cerveceros de España)

THE ENCOUNTER WITH THE RESEARCH TEAM FROM UPV

In 1999, Cerveceros de España got in contact with Dr Ángel Montoya, of the Immunotechnology Group at Valencia Polytechnic University.

“Cerveceros de España suggested that we develop a detection method that could increase specificity and reduce the detection time of lactic acid bacteria in beer. This project was of great interest to us, as it offered us the chance to give fresh impetus to the research area that we have been working on since 1993 with regard to the use of monoclonal antibodies, together with the Beer and Malt Research Association”.

Dr Ángel Montoya (Head of the research team at the Immunotechnology Group)

The Immunotechnology Group began in 1990 at Valencia Polytechnic University and its research focuses on the field of new molecular analysis systems and, in particular, biosensors. The group is based on the knowledge of professionals with a great deal of experience and international renown in the field of monoclonal antibodies, the development of methods of immunological analysis and the development of biosensors.

UPV has been carrying out research into monoclonal antibodies since 1989. In that time the Immunotechnology Group has developed over 200 antibodies and has conducted research projects into biosensors in collaboration with the food and agriculture industry, among other sectors, thus contributing to UPV’s work in general by providing solutions for this industrial sector (see Appendix 2: Technological Trends in the Food and Agriculture Industry: Research areas developed at UPV).

THE RESEARCH PROJECT

The first research project conducted at UPV on the application of monoclonal antibodies in the beer industry began in 1993, within the framework of a Collaboration Agreement signed with the Beer and Malt Research Association (see Appendix 3: Standard Agreements and Contracts for establishing relations between UPV and companies and institutions). During the following years, the research continued through agreements with various

Beer packaging line

Researcher of the Immuno-technology Group at UPV.


different companies, and in 1999 it was given a final boost when a two-year Agreement was signed with the Cerveceros de España association.

“Right from the start, our main concern has been to direct our research towards achieving tools that respond to the real working environment and dynamics of brewery laboratories. We therefore decided to work together with their researchers and created a joint programme of work that turned out to be very rewarding and productive.”

Dr Ángel Montoya (Head of the research team of the Immunotechnology Group at UPV)

The project was carried out by a mixed team comprising staff from the companies within the Cerveceros de España association and researchers of the Immunotechnology Group. The brewery groups provided around 20 strains of the most characteristic bacteria that they were most interested in recognising. UPV, for its part, developed and optimised the immunological tests and the immunosensors, which were then validated by the companies.

The total cost of the research was 318,000 €, approximately 40% of which was covered by public grants (see Appendix 4: Grants for corporate R&D, management bodies, grant schemes and financing).

This collaboration led to the development of a method for identifying lactic acid bacteria in beer, based on the use of monoclonal antibodies associated to luminescent markers. This identification method reduces by 3/4 the time needed for the microbiological control of beer.

Although this technology involves purchasing specific equipment and materials with a high market value, its use is perfectly viable; the cost of producing the antibodies is about 1000 € / mg, and approximately 10,000 tests can be conducted with every milligram, meaning that there is a unit cost of 7 € per test.

OTHER APPLICATIONS OF RAPID MOLECULAR ANALYSIS SYSTEMS, PARTICULARLY BIOSENSORS, IN THE FOOD AND AGRICULTURE INDUSTRY

One of the most strategically important areas of innovation for the European food and agriculture industry is improving the safety and quality of foodstuffs. Both the EU’s 7th framework programme and the 2004-2007 National Science and Technology Plan therefore give priority to the development of molecular systems of detection, analysis and diagnosis that are fast, sensitive, specific and that can be easily incorporated into production chains.

These molecular analysis systems are known as biosensors. The technology for designing and producing biosensors has undergone substantial advances in recent years, owing mainly to investments in companies in the biohealth, pharmaceutical and military sectors, but it has now reached a stage where it can gradually spread into other sectors such as the environmental and food and agriculture sectors.

A biosensor is a compact analysis device that consists of a biological (nucleic acid, enzyme, antibody, receptor, tissue, cell) or biomimetic recognition element linked to a transduction system that makes it possible to process the signal produced by the interaction between the recognition element and the target molecule.

Analysis of lactic acid bacteria using the system developed: left: an uncontaminated sample, right: a contaminated sample.

Diagram of the detection system developed


In the food and agriculture industry it is of particular interest in food composition analysis (characterisation and verification of its composition, determination of the degree of freshness), food safety (detection of contaminant compounds, allergens, antinutrients, toxins and pathogenic microorganisms) and continuous process control (see Appendix 1: Applications of biosensors in the food and agriculture industry).


APPENDIX 1 APPLICATIONS OF BIOSENSORS IN THE FOOD AND AGRICULTURE INDUSTRY

• Detection of L-amino acids in milk and fruit juice.

• Detection of ethanol in beer, wine and other alcoholic drinks.

• Detection of glycerol in wine.

• Detection of cholesterol in butter, lard and egg.

• Detection of citric acid in fruit juice and sports drinks.

• Detection of folic acid in fortified foods.

Determination of the degree of freshness and the shelf life of foodstuffs

• Oils: detection of polyphenols and short-chain fatty acids for evaluation of rancidity.

• Seafood: detection of ornithine and amines to determine freshness.

• Fish: detection of amines, histamine and hypoxanthine to determine freshness.

• Meat: detection of lactic acid to determine freshness.

Determination of the degree of ripeness of fruit

• Detection of glucose, sucrose and isocitrate.

Determination of the degree of wine spoilage

• Detection of 2,4,6 trichloroanisole.

PROCESS CONTROL

• Detection of glucose in fermentation processes.

• Detection of alcohol and glycerol in alcohol fermentation processes.

• Detection of lactic acid in cheese production.

FOOD SAFETY

Detection of products external to foodstuffs that are not synthesised by living organisms

• Detection and quantification of additives (sweeteners and preservatives).

• Detection of drugs.

• Detection and quantification of pesticide and fertiliser residues.

• Detection of other types of organic and heavy metal contaminants.

Detection of compounds in foodstuffs that might cause problems for the organism

• Detection of antinutritional and allergenic compounds.

Detection of biotoxins that might be related to food poisoning

• Detection of toxins of different origins: bacteria, mycotoxins, marine toxins.

Detection of pathogenic microorganisms • Detection of pathogenic bacteria and

enteric parasites.

• Detection of viruses that affect crops and livestock.

QUALITY CONTROL

Verification of food composition

• Detection of glucose in must, wine, fruit juice, soft drinks, honey, milk and yoghurt.

• Detection of fructose in fruit juice, honey, milk, jellies and artificial sweeteners.

• Detection of lactose in milk.

• Detection of lactate in cider and wine.

• Detection of starch in wheat flour.


APPENDIX 2 TECHNOLOGICAL TRENDS IN THE FOOD AND AGRICULTURE INDUSTRY (*) RESEARCH AREAS DEVELOPED AT UPV.

APPENDIX 1

(*) Source: written using the publication: Agroalimentación: tendencias tecnológicas a medio y largo plazo. 2002; Observatorio de Prospectiva Tecnológica Industrial (Food and agriculture: medium- and long-term technological trends. 2002; Industrial Technology Foresight Observatory). Ministry of Science and Technology.

FOOD QUALITY AND SAFETY

Widespread use of sensors in the control of processes

• Development of immunosensors for microbiological control of food products.

• Development of DNA matrices for early detection of pathogenic agents.

• Development of immunochemical sensors for detection of pesticide residues.

Determination of internal parameters by non-destructive technologies

• Use of ultrasound for characterisation of dairy products.

• Use of ultrasound for detection of foreign bodies in fluids.

Traceability and control of processes • Hazard analysis and critical control points

in the Food and Agriculture Industry. • Development of traceability systems and

shelf-life studies for food products. • Bromatology and microbiology of food

products.

PROCESS INNOVATION

Salting • Salting of meat and fish products by

immersion in brine.

• Simultaneous salting and defrosting of meat and fish products.

• Vacuum impregnation, use in salting cheese, fish and meat products.

• Fish desalting. • Production of high quality salted food

products with a low sodium content.

Extraction: • Supercritical fluid extraction technologies.

Drying and dehydration • Osmotic dehydration of fruit and

vegetables. • Cryoprotection of fruit by osmotic

dehydration and vacuum impregnation.

• Drying of food products using combined methods: air-microwave-vacuum.

Membrane processes • Use of nanofiltration, inverse osmosis and

pervaporation in the food industry.

Traditional product innovation

• New Technologies for the production of turron.

Vacuum processing technologies

• Vacuum impregnation and cooking; use in crystallising fruit and in producing jams at low temperatures.

Modelling and simulation

• Prediction and measurement of the physical properties of food products.

• Analysis and Modelling of mass-transfer processes.

PRESERVATION TECHNIQUES

Active packaging

• Development of functional waxes and edible films for fresh fruit.

• Use of zeolites for the elimination of ethylene in packs of fresh-cut produce.

PRODUCT INNOVATION

Functional food products

• Use of Matrix engineering in the development of functional food products.

New products

• Colloidal product technology: emulsions, foams, gels, etc.

• Use of stabilisers and emulsifiers in the formulation of food products.

SUSTAINABILITY AND LIFECYCLE

Waste Evaluation

• Characterisation and evaluation of organic waste for agricultural purposes.

• Characterisation and use of solid waste as building materials.

Recovery

• Treatment of brine from the food and agriculture industry.


APPENDIX 3 STANDARD AGREEMENTS AND CONTRACTS FOR ESTABLISHING RELATIONS BETWEEN

UPV AND COMPANIES AND INSTITUTIONS

1. Framework collaboration agreement: This consists of a declaration of intentions for future collaboration in different fields and forms, it establishes the general terms of cooperation and, in certain cases, the parties agree on a Monitoring and Planning Committee for the activities.

This type of cooperation may include granting the temporary of use of areas and facilities belonging to UPV at the request of the company in order to establish a permanent research activity on Campus for its own private use.

2. Agreement for a Research and Development project: This includes the terms in which a company contracts UPV to carry out an R&D project. It specifies the aims of the project, the work plan, the intermediate milestones to be achieved, the expected results, the necessary resources and the contributions to be made by each party, and it establishes a confidentiality regime and economic conditions. The ownership of the results is negotiated for each case according to the contributions made my each party.

3. Patent and software licence agreements: This includes the terms in which a company or institution acquires the industrial rights of a certain technology owned by UPV. The exclusivity terms, geographical limits and duration of the licence are negotiated for each case according to the plan for use of the results and the economic conditions of the agreement.

4. Technological Support and Advice Agreement: This consists of carrying out technical laboratory work leading to the application of knowledge acquired by researchers at UPV to achieve new products or processes without this work involving a research activity. The structure and terms of this type of agreement are similar to those of the Research and Development agreement.

5. Technical services agreement: This consists of rendering a technical service (measurement, report) or selling a product manufactured directly by the University as a consequence of its R&D activity.

6. Training: Training provided by UPV can consist of attending general courses offered by UPV through the Post-graduate Training Centre or requesting a custom-designed course.

7. Work placements for students in companies: This establishes the conditions in which students of UPV can spend a period in a company on a work placement scheme. The work assigned to the student must have educational value and be of use to the company. The student will receive a grant from the company, without this involving any employment relationship. There must be a tutor in the company and at the University to supervise the work.

8. Recruitment of university graduates: The UPV-FSVE Recruitment Agency is in charge of finding suitable candidates for job offers received by UPV.


APPENDIX 4 GRANTS FOR CORPORATE R&D; MANAGEMENT BODIES, GRANT SCHEMES AND FINANCING

Regional: Institute of Valencian Small and Medium-sized Industries, IMPIVA. http://www.impiva.es

Company support programmes.

National: Ministry of Industry, Trade and Tourism http://www.mityc.es

Programme for the Promotion of Technical Research, PROFIT. Ministry of Education and Science http://www.mec.es.

Grants and Aid for Science and Technology: PETRI programme for the promotion of technical research; Unique and Strategic Projects; R&D Projects.

Torres Quevedo programme for promoting the placement of doctors and technologists in companies.

Centre for Industrial Technological Development http://www.mec.es

Financing of corporate R&D projects. Support for the creation of new technology-based companies NEOTEC

https://www.neotec.cdti.es/

European: European Commission: EU Framework Programme for Research and Technological Development. http://europa.eu.int/comm/research/fp6/pdf/blue_guide_es.pdf

APPENDIX 5 TAX TREATMENT OF R&D

Tax Deductions for R&D are the strongest financing framework for innovation activities in Spanish companies. They can be applied by means of reductions in the total tax payable by companies depending on how much they invest in R&D during each financial year and depending on the characteristics of the activity undertaken with said investment according to a classification of Technological Research, Development and Innovation projects. Companies may decide to present their own declaration justifying their expenditure on R&D, or they may request that an organisation accredited by the Spanish Accreditation Agency (ENAC) issues a reasoned report certifying the expenditure, classifying the project and specifying the type of deduction that is applicable. This type of report is binding. The regulatory framework consists of Legislative Royal Decree 4/2004 of 5 March (RDL 4/2004 de 5 de marzo), which approved the revised text of the Corporate Income Tax Law (Ley del Impuesto sobre sociedades), and Royal Decree 1432/2003 of 21 December (RD 1432/2003 de 21 de diciembre) on the regulation of reasoned reports.

bere caso cerveceros espana

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