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Analysis of grapes and the first stages of the vinification process in wine contamination with Brettanomyces bruxellensis

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Abstract

Brettanomyces bruxellensis is a major cause of wine spoilage due to the production of ethyl phenols, and it has become a major worldwide oenological concern in recent years. The most critical factor in volatile phenol production is the presence of microorganisms responsible for biosynthesis. In this work, carried out during three consecutive harvests, grapes and the first step in grape processing (stemming–crushing) have been evaluated as the origin of wine contamination by these spoilage yeasts. Results showed that there was nil or minimal presence of Brettanomyces yeasts in grapes and on the stemmer, in levels that the method was not able to detect. This shows that the main contamination of wines by this microorganism occurs in later stages of the vinification process and/or during storage. The contamination of many wines with Brettanomyces, either via the fruit or from the winery environment, during the early stages of vinification and before the start of aging, was confirmed by analyzing 100 recently made red wines, in which this yeast was detected in a high percentage of wines (27 %). However, the level of Brettanomyces yeasts found in the samples was low, with values which would not be sufficient to cause organoleptic defects. Consequently, this study confirms that many wines are still tainted by Brettanomyces when the winemaking phase comes to an end. It was also found that wines with problems during fermentation had a higher level of Brettanomyces. This is why it is essential to ensure strict controls during fermentation processes and conservation to prevent wine becoming spoiled or tainted.

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References

  1. Deák T, Beuchat LR (1993) Use of indirect conductimetry for predicting growth of food spoilage yeasts under various environmental conditions. J Ind Microbiol 12:301–308

    Article  Google Scholar 

  2. Thomas DS (1993) Yeast as spoilage organisms in beverages. In: Rose AH, Harrison JS (eds) The yeast vol 5: yeast technology, 2nd edn. Academic Press, London

    Google Scholar 

  3. Loureiro V, Malfeito-Ferreira M (2006) Food Spoilage Microorganisms. In: Blackburn C (ed) Spoilage activities of Dekkera/Brettanomyces spp. Woodhead Publishing Limited, Cambridge

    Google Scholar 

  4. Chatonnet P, Dubourdieu D, Boidron JN, Pons M (1992) The origin of ethylphenols in wines. J Sci Food Agric 60:165–178

    Article  CAS  Google Scholar 

  5. Chatonnet P, Dubordieu D, Boidron J (1993) Synthesis of volatile phenols by Saccharomyces cerevisiae in wines. J Sci Food Agric 62:101–202

    Article  Google Scholar 

  6. Loureiro V, Malfeito-Ferreira M (2003) Spoilage yeasts in the wine industry. Int J Food Microbiol 86(1–2):23–50

    Article  CAS  Google Scholar 

  7. Gerbaux V, Naudin R, Meurgues O, Monamy C (2000) Etude des phénols volatils dans les vins de Pinot noir en Bourgogne. Bull del´OIV 73(835–836):581–599

    CAS  Google Scholar 

  8. Wedral D, Shewfelt R, Frank J (2010) The challenge of Brettanomyces in wine. LWT Food Sci Technol 43:1474–1479

    Article  CAS  Google Scholar 

  9. Suárez R, Suárez-Lepe JA, Morata A, Calderón F (2007) The production of ethylphenols in wine by yeasts of the genera Brettanomyces and Dekkera. A review. Food Chem 102:10–21

    Article  Google Scholar 

  10. Silva P, Cardoso H, Gerós H (2004) Studies on the wine spoilage capacity of the Brettanomyces/Dekkera spp. Am J Enol Vitic 55(1):65–72

    CAS  Google Scholar 

  11. Mortimer R, Polsinelli M (1999) On the origin of wine yeast. Res Microbiol 150:199–204

    Article  CAS  Google Scholar 

  12. Renouf V, Lonvaud-Funel A (2007) Development of an enrichment medium to detect Dekkera/Brettanomyces bruxellensis, a spoilage wine yeast, on the surface of grape berries. Microbiol Res 162:154–167

    Article  CAS  Google Scholar 

  13. Ocón E, Gutiérrez AR, Garijo P, López R, Santamaría P (2010) Presence of non-Saccharomyces yeasts in cellar equipments. Food Microbiol 27:1023–1027

    Article  Google Scholar 

  14. Pretorius IS (2000) Tailoring wine yeast for the new millennium: novel approaches to the ancient art of winemaking. Yeast 16:675–729

    Article  CAS  Google Scholar 

  15. Renouf V, Lonvaud-Funel A, Coulon J (2007) The origin of Brettanomyces bruxellensis in wines: a Review. J Int Sci Vigne Vin 41–3:161–173

    Google Scholar 

  16. Alguacil M, Fidalgo M, Jiménez J, Lozano JI, Neva MA, Perdigones F (1998) Detección de Dekkera/Brettanomyces en instalaciones de vendimia mediante PCR. Alimentos Equipos y Tecnología 10:81–85

    Google Scholar 

  17. Baleiras-Couto MM, Gomes AS, Casal M, Duarte FL (2011) Survey of yeast diversity during bottling processes using restriction analysis of 26S ribosomal DNA (rDNA). Aust J Grape Wine Res 18:39–42

    Article  Google Scholar 

  18. Renouf V, Claisse O, Lonvaud-Funel A (2007) Inventory and monitoring of wine microbial consortia. Appl Microbiol Biotechnol 75:149–164

    Article  CAS  Google Scholar 

  19. Ocón E, Garijo P, Sanz S, Olarte C, López R, Santamaría P, Gutiérrez AR (2013) Screening of yeast mycoflora in winery air samples and their risk of wine contamination. Food Control 34:261–267

    Article  Google Scholar 

  20. Connell L, Stender H, Edwards CG (2002) Rapid detection of Brettanomyces from winery air samples based on peptide nucleic acid analysis. Am J Enol Vitic 53:322–324

    CAS  Google Scholar 

  21. Fugelsang KC, Zoecklein CG (2003) Population dynamics and effects of Brettanomyces bruxellensis strains on Pinot noir (Vitis vinifera L.) wines. Am J Enol Vitic 54:294–300

    CAS  Google Scholar 

  22. Martorell P (2006) Desarrollo y aplicación de sistemas rápidos para la detección, identificación y caracterización de levaduras alterantes en alimentos. Tesis Doctoral. Universidad de Valencia, España

    Google Scholar 

  23. Prakitchaiwattana CJ, Fleet GH, Heard GM (2004) Application and evaluation of denaturing gradient gel electrophoresis to analysis the yeast ecology of wine grapes. FEMS Yeast Res 4:865–877

    Article  CAS  Google Scholar 

  24. Millet V, Lonvaud-Funel A (2000) The viable but non-culturable state of wine microorganisms during storage. Lett Appl Microbiol 30:136–141

    Article  CAS  Google Scholar 

  25. du Toit WJ, Pretorius IS, Lonvaud-Funel A (2005) The effect of sulphur dioxide and oxygen on the viability and culturability of a strain of Acetobacter pasteurianus and a strain of Brettanomyces bruxellensis isolated from wine. J Appl Microbiol 98:862–871

    Article  Google Scholar 

  26. Divol B, Lonvaud-Funel A (2005) Evidence for viable but non culturable yeasts in botrytis-affected wine. J Appl Microbiol 99:85–93

    Article  CAS  Google Scholar 

  27. Ocón E, Garijo P, Santamaría P, López R, Olarte C, Gutiérrez AR, Sanz S (2013) Comparison of culture media for the recovery of airborne yeast in wineries. Lett Appl Microbiol 57:241–248

    Article  Google Scholar 

  28. Barata A, Caldeira J, Botheleiro R, Pagliara D, Malfeito-Ferreira M, Loureiro V (2008) Survival patterns of Dekkera bruxellensis in wines and inhibitory effect of sulphur dioxide. Int J Food Microbiol 121:201–207

    Article  CAS  Google Scholar 

  29. Oelofse A, Pretorius IS, Du Toit M (2008) Significance of Brettanomyces and Dekkera during winemaking: a synoptic review. South Afr J Enol Vitic 29(2):128–144

    CAS  Google Scholar 

  30. Rodrígues N, Gonçalves G, Pereira-da-Silva S, Malfeito-Ferreira M, Loureiro V (2001) Development and use of a differential medium to detect yeast of the genera Dekkera/Brettanomyces. J Appl Microbiol 90:588–599

    Article  Google Scholar 

  31. Esteve-Zarzoso B, Belloch C, Uruburu F, Querol A (1999) Identification of yeasts by RFLP analysis of the 5.8S rRNA gene and the two ribosomal internal transcribed spacers. Int J Syst Bacteriol 49:329–337

    Article  CAS  Google Scholar 

  32. White TJ, Bruns T, Lee S, Taylor J (1990) In: Innis MA, Gelfalnd DH, Sninsky JJ, White TT (eds) PCR protocols. A guide to methods and applications. Amplification and direct sequencing of fungi ribosomal RNA genes for phylogenetics. Academic Press, San Diego

    Google Scholar 

  33. Willenburg E, Divol B (2012) Quantitative PCR: an appropriate tool to detect viable but not culturable Brettanomyces bruxellensis in wine. Int J Food Microbiol 160:131–136

    Article  CAS  Google Scholar 

  34. Barata A, Malfeito-Ferreira M, Loureiro V (2012) Changes in sour rotten grape Berry microbiota during ripening and wine fermentation. Int J Food Microbiol 154:152–161

    Article  CAS  Google Scholar 

  35. Jolly NP, Augustyn OPH, Pretorius IS (2003) The occurrence of non-Saccharomyces cerevisiae yeast species over three vintages in four vineyards and grape musts from four production regions of the Western Cape, South Africa. South Afr J Enol Vitic 24(2):35–42

    Google Scholar 

  36. Raspor P, Milek DM, Polanc J, Mozina SS, Cadez N (2006) Yeasts isolated from three varieties of grapes cultivated in different locations of the Dolenjska wine-growing region, Slovenia. Int J Food Microbiol 109:97–102

    Article  CAS  Google Scholar 

  37. Cordero-Bueso G, Arroyo T, Serrano A, Tello J, Aporta I, Vélez MD, Valero E (2011) Influence of grape farming system and vine variety on yeast communities associated with grape berries. Int J Food Microbiol 145:132–139

    Article  Google Scholar 

  38. Dias L, Prereira-da-Silva S, Tavares M, Malfeito-Ferreira M, Loureiro V (2003) Factor affecting the production of 4-ethylphenol by the yeast Dekkera bruxellensis in enological conditions. Food Microbiol 20(4):377–384

    Article  CAS  Google Scholar 

  39. Ocón E, Garijo P, Sanz S, Olarte C, López R, Santamaría P, Gutiérrez AR (2013) Analysis of airborne yeast in one winery over a period of one year. Food Control 30:585–589

    Article  Google Scholar 

  40. Conterno L, Joseph CML, Arvik TJ, Hening-Kling T, Bisson LF (2006) Genetic and physiological characterization of Brettanomyces bruxellensis strains isolated from wines. Am J Enol Vitic 57:139–147

    CAS  Google Scholar 

  41. Morneau AD, Zuehlke JM, Edwards CG (2011) Comparison of media formulations used to selectively cultivate Dekkera/Brettanomyces. Lett Appl Microbiol 53:460–465

    Article  CAS  Google Scholar 

  42. Jensen SL, Arneborg N, Edwards CG (2009) Identification and characterization of Dekkera bruxellensis, Candida pararugosa and Pichia guilliermondii isolated from commercial red wines. Food Microbiol 26:915–921

    Article  CAS  Google Scholar 

  43. Agnolucci M, Vignentini I, Capurso G, Merico A, Tirelli A, Compagno C, Foschino R, Nuti M (2009) Genetic diversity and physiological traits of Brettanomyces bruxellensis strains isolated from Tuscan Sangiovese wines. Int J Food Microbiol 130(3):238–244

    Article  CAS  Google Scholar 

  44. Barata A, Malfeito-Ferreira M, Loureiro V (2012) The microbial ecology of wine grape berries. Int J Food Microbiol 153:243–259

    Article  CAS  Google Scholar 

  45. Farina L, Boido E, Carrau F, Dellacassa E (2007) Determination of volatile phenols in red wines by dispersive liquid-liquid microextraction and gas chromatography-mass spectrometry detection. J Chromatogr A 1157:46–50

    Article  CAS  Google Scholar 

  46. Silva LR, Andrade PB, Valentao P, Seabra RM, Trujillo ME, Velázquez E (2005) Analysis of non-coloured phenolics in red wine: effect of Dekkera bruxellensis yeasts. Food Chem 89:185–189

    Article  CAS  Google Scholar 

  47. Vigentini I, Romano A, Compagno C, Merico A, Molinari F, Tirelli A, Foschino R, Volonterio G (2008) Physiological and oenological traits of different Dekkera/Brettanomyces bruxellensis straits under wine-model conditions. FEMS Yeast Res 8:1087–1096

    Article  CAS  Google Scholar 

  48. Renouf V, Lonvaud-Funel A (2005) Effet microbiologique de l´usage de barriques neuves et/ou de barriques anciannes. Rev Fr Oenol 211:10–14

    Google Scholar 

  49. Serpagi V, Remize F, Recorbet G, Gaudot-Dumas E, Sequeira-Le Grand A, Alexandre H (2012) Characterization of the “viable but non culturable” (VBNC) state in the wine spoilage yeast Brettanomyces. Food Microbiol 30:438–447

    Article  Google Scholar 

  50. Laforgue R, Lonvaud-Funel A (2012) Hydroxycinnamic acid decarboxylase activity of Brettanomyces bruxellensis involved in volatile phenol production: relationship with cell viability. Food Microbiol 32:230–234

    Article  CAS  Google Scholar 

  51. Renouf V, Falcou M, Miot-Sertier C, Perello MC, De Revel G, Lonvaud-Funel A (2006) Interactions between Brettanomyces bruxellensis and other yeast species during the initial stages of winemaking. J Appl Microbiol 100(6):1208–1219

    Article  CAS  Google Scholar 

  52. Uscanga MG, Delia ML, Strehaiano P (2003) Brettanomyces bruxellensis: effect of oxygen on growth and acetic acid production. Appl Microbiol Biotechnol 61(2):157–162

    Article  Google Scholar 

  53. Kheir J, Salamech D, Strehaiano P, Brandam C, Lteif R (2013) Impact of volatile phenols and their precursors on wine quality and control measures of Brettanomyces/Dekkera yeasts. Eur Food Res Technol 237:655–671

    Article  CAS  Google Scholar 

  54. Zuehlke JM, Petrova B, Edwards CG (2013) Advances in the control of wine spoilage by Zygosaccharomyces and Dekkera/Brettanomyces. Annu Rev Sci Technol 4:57–78

    Article  CAS  Google Scholar 

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Acknowledgments

This study has been undertaken with a Grant from the Government of La Rioja, Spain (Project “PR-08-12,” Project “PR-07-13”) and from La Rioja University-Banco Santander (Project PROFAI 13/02).

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None.

Compliance with Ethics Requirements

This article does not contain any studies with human or animal subjects.

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Authors and Affiliations

  1. ICVV, Instituto de Ciencias de la Vid y el Vino, Universidad de La Rioja, Gobierno de La Rioja, CSIC, C/Madre de Dios 51, 26006, Logroño, Spain

    P. Garijo, L. González-Arenzana, I. López-Alfaro, T. Garde-Cerdán, R. López, P. Santamaría & A. R. Gutiérrez

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  1. P. Garijo
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  2. L. González-Arenzana
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  3. I. López-Alfaro
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  5. R. López
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  6. P. Santamaría
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  7. A. R. Gutiérrez
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Correspondence to A. R. Gutiérrez.

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Garijo, P., González-Arenzana, L., López-Alfaro, I. et al. Analysis of grapes and the first stages of the vinification process in wine contamination with Brettanomyces bruxellensis . Eur Food Res Technol 240, 525–532 (2015). https://doi.org/10.1007/s00217-014-2351-4

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  • DOI: https://doi.org/10.1007/s00217-014-2351-4

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