Comparison of Brettanomyces yeast presence in young red wines in two consecutive vintages


Two hundred young wines from the D.O.Ca. Rioja (Spain) coming from two consecutive vintages, 100 wines from 2012 and 100 wines from 2013, were analysed and compared with respect to physicochemical composition and Brettanomyces presence. This microorganism was tested using two techniques: qPCR and an odorimetric test. In 2013, the harvest was characterized by late ripening, health problems with the grapes, and difficulties during vinification. Differences between 2012 and 2013 wines were found both for analytical parameters and as for the presence of the spoilage yeast, Brettanomyces, which was much more prevalent in the 2013 vintage. Moreover, the results showed that for some wines in which Brettanomyces was detected by qPCR at concentration higher than 102 cells/ml, no “Brett” character was found in the odorimetric test. This would indicate that the contamination of a wine by this yeast is not the only factor involved in spoilage. The strains of Brettanomyces, their physiological state, and viability, besides the specific composition of wine, could be important in the “Brett” character appearance. In this study, the wines which developed spoilage in the odorimetric test had slightly lower pH and alcoholic strength values, but higher levels of volatile acidity, reducing sugars and density in comparison with those wines than were not affected. Most of these factors are favourable for the development of Brettanomyces.

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  1. 1.

    Agnolucci M, Vigentini I, Capurso G, Merico A, Tirelli A, Compagno C, Foschino R, Nuti M (2009) Genetic diversity and physiological traits of Brettanomyces bruxelliensis strain isolated from Toscan Sangiovese wines. Int J Food Microbiol 3:238–244

    Article  Google Scholar 

  2. 2.

    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 

  3. 3.

    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

    CAS  Article  Google Scholar 

  4. 4.

    Chandra M, Madeira I, Coutinho A, Alvergaria H, Malfeito-Ferreira M (2016) Growth and volatile phenol production by Brettanomyces bruxellensis in different grapevine varieties during fermentation and in finished wine. Eur Food Res Technol 242:487–494

    CAS  Article  Google Scholar 

  5. 5.

    Commission Regulation (ECC) Nº 2676/90 of 17 September 1990 determining community methods for the analysis of wines. Off J Eur Commun L272, 33 of 3 Oct 1990. (

  6. 6.

    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 

  7. 7.

    Coulon J, Perello MC, Lonvaud-Funel A, De Revel G, Renouf V (2010) Brettanomyces bruxellensis evolution and volatile phenols production in red wines during storage in bottles. J Appl Microbiol 108:1450–1458

    CAS  Article  Google Scholar 

  8. 8.

    Couto JA, Barbosa A, Hogg T (2005) A simple cultural method for the presumptive detection of the yeast Brettanomyces/Dekkera in wines. Lett Appl Microbiol 41:505–510

    CAS  Article  Google Scholar 

  9. 9.

    Dias L, Pereira-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 20:377–384

    CAS  Article  Google Scholar 

  10. 10.

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

    CAS  Article  Google Scholar 

  11. 11.

    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 

  12. 12.

    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 

  13. 13.

    Garijo P, González-Arenzana L, López-Alfaro I, Garde-Cerdán T, López R, Santamaría P, Gutiérrez AR (2015) Analysis of grapes and the first stages of vinification process in wine contamination with Brettanomyces bruxellensis. Eur Food Res Technol 240:525–532

    CAS  Article  Google Scholar 

  14. 14.

    Gervaux V, Vincent B, Bertrant A (2002) Influence of maceration temperature and enzymes on the content of volatile phenols in Pinot Noir wines. Am J Enol Vitic 53:131–137

    Google Scholar 

  15. 15.

    Hierro N, Esteve-Zarzoso B, González A, Mas A, Guillamón JM (2006) Real-time quantitative PCR (QPCR) and reverse transcription-QPCR for detection and enumeration of total yeasts in wine. Appl Environ Microbiol 72:7148–7155

    CAS  Article  Google Scholar 

  16. 16.

    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

    CAS  Article  Google Scholar 

  17. 17.

    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

    CAS  Article  Google Scholar 

  18. 18.

    Licker JL, Acree TE, Heníck-Kling T (1998) What is “Brett” (Brettanomyces) flavor? a preliminary investigation. In: Waterhouse AL, Ebeler SE (eds) Chemistry of wine flavor. Americam Chemical Society, Washington

    Google Scholar 

  19. 19.

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

    CAS  Article  Google Scholar 

  20. 20.

    Martorell P, Querol A, Fernández-Espinar MT (2005) Rapid identification and enumeration of Saccharomyces cerevisiae cells in wine by real-time PCR. Appl Environ Microbiol 71:6823–6830

    CAS  Article  Google Scholar 

  21. 21.

    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

  22. 22.

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

    CAS  Article  Google Scholar 

  23. 23.

    Mira de Orduña R (2010) Climate change associated on grape and wine quality and production. Food Res Int 43:1844–1855

    Article  Google Scholar 

  24. 24.

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

    CAS  Google Scholar 

  25. 25.

    Phister TG, Rawsthorne H, Joseph CML, Mills DA (2007) Real-time PCR assay for detection and enumeration of Hanseniaspora species from wine and juice. Am J Enol Vitic 58:229–233

    CAS  Google Scholar 

  26. 26.

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

    CAS  Article  Google Scholar 

  27. 27.

    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 

  28. 28.

    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 100:1208–1219

    CAS  Article  Google Scholar 

  29. 29.

    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. Microbial Res 162:154–167

    CAS  Article  Google Scholar 

  30. 30.

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

    CAS  Google Scholar 

  31. 31.

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

    CAS  Article  Google Scholar 

  32. 32.

    Salinas F, Garrido D, Ganga A, Veliz G, Martínez C (2009) Taqman real-time PCR for the detection and enumeration of Saccharomyces cerevisiae in wine. Food Microbiol 26:328–332

    CAS  Article  Google Scholar 

  33. 33.

    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 

  34. 34.

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

    CAS  Google Scholar 

  35. 35.

    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 

  36. 36.

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

    CAS  Article  Google Scholar 

  37. 37.

    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

    CAS  Article  Google Scholar 

  38. 38.

    Woolfit M, Rozpedowska E, Piskur J, Wolfe KH (2007) Genome survey sequencing of the wine spoilage yeast Dekkera (Brettanomyces) bruxellensis. Eukaryot Cell 6:721–733

    CAS  Article  Google Scholar 

  39. 39.

    Zott K, Claisse O, Lucas P, Coulon J, Lonvaud-Funel A, Masneuf-Pomarede I (2010) Characterization of the yeast ecosystem in grape must and wine using real-time PCR. Food Microbiol 27:559–567

    CAS  Article  Google Scholar 

  40. 40.

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

    CAS  Article  Google Scholar 

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This study has been undertaken with grants from the Government of La Rioja, Spain (Project “PR-08-12”, Project “PR-07-13”), from La Rioja University-Banco Santander (Project PROFAI 13/02), and from INIA (Project RTA 2013-00053-C03-03). Authors would like to thank Victor Llop Santamaría for excellent technical assistance.

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Correspondence to Susana Sanz.

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Garijo, P., Gutiérrez, A.R., López, R. et al. Comparison of Brettanomyces yeast presence in young red wines in two consecutive vintages. Eur Food Res Technol 243, 827–834 (2017).

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  • Red wines
  • Spoilage
  • “Brett” character
  • Harvest conditions