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Università Cattolica del Sacro Cuore PIACENZA |
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(OTA) Management in Grapes and Wine
Contract n. QLK1-CT-2001-01761
Risk Assessment and Integrated Ochratoxin A (OTA) Management in Grapes and Wine |
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| Summary year 3 (72 kB) | ||
| III Annuncement (PDF 1,2 MB) | ||
| Program (PDF 760 KB) | ||
| WINE OCHRA-RISK | QLK1-CT-2001-01761 | D E S C R I P T I O N |
| Background |
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| The world-wide occurrence of OTA contamination of raw agricultural products has been amply documented in a variety of plant products and recently OTA has been detected also in wine and grape juice. Fungi responsible for OTA presence were studied especially on cereals where Penicillium verrucosum and A. ochraceus are considered the main producers. | ||||||||||||||
| Few studies carried out in EU countries on grape have shown that there is a great variability and a significant morphological and toxicological difference among Aspergillus and Penicillium species occurring on grapes; species associated with cereals seem absent. | ||||||||||||||
| Surveys carried out on wine showed a higher OTA concentration in red versus white and rosé wines. A correlation with the latitude of the production region was also demonstrated: the further south the provenance, the more frequent the occurrence and the greater the concentration of OTA. | ||||||||||||||
| Wine contribution to mean daily OTA intake cannot be considered negligible: wine alone could supply the diet of an average drinker with an OTA amount equal or even superior to the TDI recommended by the Scientific Committee on Food of the European Commission. | ||||||||||||||
Over-all objective |
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| Assessment of risk of ochratoxin A (OTA) in grape and wine in Europe and protection of the consumer's health by decreasing the amount of toxin with the aid of integrated management of production and processing. | ||||||||||||||
Expected achievements |
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| Description of work |
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| Six European countries and Israel will be included in the project. Twelve groups will contribute to the study, 10 are expert researchers in the field of viticulture, fungal ecology/physiology/ taxonomy, food microbiology, analytical chemistry, diagnostics, food technology, oenology, epidemiology and statistics, while the last 2 guarantee the connection between scientific team and the final users. | ||||||||||||||
| The workplan is divided into five major tasks of which the first two consist of data acquisition to define the CCP and factors able to influence them. The third one defines preventive and corrective actions for vineyards and wine clubs, whose effect can be efficiently controlled (Task 4). Task 5, inclusive of all the others, defines the real risk for OTA presence in grape and wine and gives recommendations for the integrated management of grape production and processing aiming at reducing OTA risk as much as possible. | ||||||||||||||
Expected results |
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| Objectives: |
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aim of this project is to assess the occurrence of ochratoxin A (OTA)
in grape and wine and to reduce the intake of OTA from grapes and wine.
It is pursued using the HACCP approach, by identifying the key elements
for OTA in grape and wine, and providing tools for preventive and
corrective action, according to an integrated approach. The research steps, planned for the second year, consists of collecting data on:
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Results and Milestones: |
| Trans-European populations of OTA producing fungi in grapes were
isolated following a common protocol; they were identified and
characterised for OTA production. Aspergillus section Nigri were dominant as potential OTA producing fungi in the vineyard; in particular, A. carbonarius seems confirmed as the target pathogen, since especially because most of its strains were able to produce large amounts of the toxin. Initial results regarding ecological studies, based on different species belonging to Aspergillus section Nigri, suggest that A. carbonarius is able to grow over the water activity (aw) range from 0.88 to 0.99 aw at 25 and 35 °C, but not at 15 °C. There is a statistically significant effect of aw x strain, and also aw x temperature. There seems to be a difference in growth between strains of A. carbonarius from the same country. The aw range for all members of the Aspergillus section Nigri were similar at steady-state temperatures. Optimal growth of all species/strains were around 0.98 aw, and decreased progressively down to 0.90-0.88 aw. Molecular studies are in progress to characterize the ochratoxigenic species occurring on grapes in Europe. The preliminary data of AFLP analysis performed on 300 strains among Aspergillus section Nigri representatives of identified species and sampled territory showed a clear pattern's peaks for the three species main occurring on grapes (A. japonicus, A. carbonarius and A. niger aggregate). Sequencing variable DNA regions (ITS1/ITS2, 28S and calmodulin) of 30 strains belonging the three groups showed specific regions useful for designing specific primers for A. carbonarius species and A. japonicus/aculeatus groups. Also RFLP analysis is in progress. RFLP and AFLP analysis performed on A. niger aggregate are able to separate A. niger aggregate in two groups (N and T). All OTA producers strains belong to pattern N. Studies carried out during wine making have underlined that during maceration OTA content in must increases while a reduction was observed after malolactic fermentation. Reduction of the amount of OTA during wine making can be obtained by the addition of products containing charcoal. The addition of these adsorbents to red wine up to 20 g/hl does not modify substantially the colour, but high amounts of adsorbent (≥ 50 g/hl), even with the combination of oenological tannins, results in significant changes in the phenolic components of red wine. Interesting results were also obtained with the addition of bacteria, in particular Lactobacillus plantarum, or enzymes (papain and acid protease P3), which can degrade OTA in red must. Storage of table grapes did not increase the number of OTA producing fungi that could be isolated from the berries; in fact, ochratoxigenic fungi associated with grape decreased either after storage for 7 days or for 30 days at 0°C. Protocols for field sampling, testing isolates for OTA production, and OTA analysis in grapes and wine were prepared and validated. A sampling protocol was developed for collection of bunches in the field, in order to estimate OTA content in vineyards. A data base is in progress with information about vineyards characteristics and farming methods. It is based on data collected using a questionnaire developed by the partners. Maps are also available with all places sampled georeferenced. Distribution maps were prepared for black aspergilli. |
Benefits and Beneficiaries: |
| Ochratoxin
A (OTA) is a toxin, considered to be a genotoxic carcinogen, and
therefore contaminated foods pose considerable risk to the consumer.
Exposure to OTA should be reduced as much as possible. The aim of this multidisciplinary project is to assess the occurrence and reduce the intake of OTA from grapes and wine. Rapid detection methods for monitoring OTA (ELISA) and for the fungi producing OTA (ELISA and PCR) will be considered and confirmed by HPLC and morphotaxonomy. The project characterizes the entire food chain, from the field to the final processed product. Extension workers, and producers involved in wine production will also be included to guarantee feedback of results obtained and dissemination of results to final end-users. |
Future Actions (if applicable): |
| Field
sampling and characterisation of strains collected from grape will
continue as well as the collection of data about farming methods and
climate in order to determinate CCPs and identify all the possible
preventive and corrective actions during the wine-making process. This
will lead to the establishment of a monitoring system able to confirm
the assessed risk of fungal and OTA presence. All data collected in field and in ecological trials in vitro will allow the development of a DSS. |
Risk Assessment and Integrated Ochratoxin A (OTA) Management in Grapes and Wine |
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| WINE OCHRA-RISK | QLK1-CT-2001-01761 | P A R T N E R S |
| Co-ordinator | |
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Paola BATTILANI Catholic University Piacenza, Faculty of Agriculture, Italy tel: +39/0523/599254 fax: +39/0523/599256 |
| Partners | |
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Javier CABAÑES Facultat de Veterinaria, Universitat Autonoma de Barcelona, Spain tel: +34/9358/11749 - 34 fax: +34/9358/12006 |
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Zofia LAWRENCE CABI Bioscience, United Kingdom tel: +44/1491/829061 fax: +44/1491/829100 |
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Armando VENANCIO Departamento Engenharia Biologica, Universidade do Minho, Portugal tel: +351/253/604413 fax: +351/253/678986 |
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Giusy MULE' Istituto di Scienze delle Produzioni Alimentari (ISPA), CNR, Italy tel: +39/080/5912830 fax: +39/080/5486063 |
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Eleftherios TJAMOS Agricultural University of Athens, Faculty of Crop Sciences and Production, Greece tel: +30/152/94505 fax: +30/152/94509 |
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Amnon LICHTER Department of Postharvest Science, ARO The Volcani Center, Israel tel: +972/3/9683684 fax: +972/3/9683622 |
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Naresh MAGAN Cranfield Biotechnology Centre, Cranfield University, United Kingdom tel: +44/0123/4754339 fax: +44/123/4750907 |
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Vicente SANCHIS Food Technology Department, Universitat de Lleida, Spain tel: +34/973/702535 fax: +34/973/702596 |
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Ahmed LEBRIHI Ecole Nationale Supérieure Agronomique de Toulouse, France tel: +33/05/62/193944 fax: +33/05/62/193901 |
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Giordano ZINZANI CAVIRO, Società Cooperativa s.r.l., Italy tel: +39/543/775519 fax: +39/543/720353 |
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Santiago MINGUEZ Jefe del Servicio de Viticultura i Enologia, Institut Catalá de la Vinya i el Vi, Spain tel: +34/93/8900211 fax: +34/93/8900354 |
| EC Contact | |
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Achim BOENKE Scientific Officer, European Commission tel: +32/2/2960756 fax: +32/2/2964322 |
Risk Assessment and Integrated Ochratoxin A (OTA) Management in Grapes and Wine |
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| WINE OCHRA-RISK | QLK1-CT-2001-01761 | R O L E O F P A R T N E R S |
| Role and contribution of Partners |
| Most of the partners will contribute to WP1 and WP2 (P1, P2, P4, P6, P7, P9, P10). They all have the competence to manage fungal isolation, characterisation for ochratoxin production and identification at genera level, so as to prepare a suitable questionnaire for collecting field data. |
| All of the partners will be supported for Aspergillus and Penicillium identification at species level by an Authority in this field, P3. Besides, the work of the groups will be improved by P2 and P5, which are experts in molecular techniques, and they will manage genetic characterisation to descibe population genetics and to obtain rapid detection techniques. |
| WP3 is specific for Ecology and it will be managed by P8 and P9 which are expert in this issue, so as in modeling data obtained in these experiments to describe fungal behaviour in different environmental conditions. They will be partially supported by P6, as expert in biological control, and by P7, as expert in table grape. They will also have exchanges with P1 who will use data from this WP as input in others, especially WP9. |
| WP5 is the application in field of results obtained in WP3: prevenctive actions to reduce OTA content in grape. It will be guided by P6, but field trials will also be carried out in other countries with the help of the subcontractor of P7, expert in field trials, and P11 and P12, which represent the connection between scientific team and the final users. |
| P4, expert in winemaking, is responsible of WP4, where corrective actions to reduce OTA content in grape after harvesting will be studied. He will cooperate with P1, which also include an expert in winemaking, and with P7, expert in table grape. They will also manage WP6, were the results obtained will be applied during vinification, also supported by P10, expert in winemaking from a different country, P11 and P12 which represent the connection with the final users also for winemaking. |
| P1 is expert in application of statistics to phytopathological data and it will manage WP8 and WP9, which concern the assessment of risk for OTA presence in grape and wine and the elaboration of a Decision Support System (DSS) for the integrated management of OTA in the grape food chain. As just defined in the general comments, the contribute of all the partners will be fundamental. |
| P1 will also be responsible for WP7, in which sampling protocols for monitoring OTA producing fungi and OTA will be prepared, taking into account Critical Control Points (CCP). All the partners will take part in this WP with suggestions, but especially P9, as responsible of WP2, where CCP will be defined, and P5, as expert in analytical methods, will support P1. A more active role will be supported by P11 and P12 in checking sampling protocols. |
Risk Assessment and Integrated Ochratoxin A (OTA) Management in Grapes and Wine |
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| WINE OCHRA-RISK | QLK1-CT-2001-01761 | L I N K S |
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European Commission |
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| European Commission - Belgium www.cec.eu.int |
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Relevant links on mycotoxins |
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Risk Assessment and Integrated Ochratoxin A (OTA) Management in Grapes and Wine |
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| WINE OCHRA-RISK | QLK1-CT-2001-01761 | L I T E R A T U R E |
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