Detection of <i>Brettanomyces</i> spp. in Red Wines Using Real‐Time PCR

Tofalo, Rosanna; Schirone, Maria; Corsetti, Aldo; Suzzi, Giovanna

doi:10.1111/j.1750-3841.2012.02871.x

Cited by 32 publications

(24 citation statements)

References 41 publications

(50 reference statements)

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“…qPCR can even be multiplexed to detect a number of organisms in one assay (Selma et al, 2009). This technique has been developed to detect and quantify total yeasts (Hierro et al, 2006a), Brettanomyces (Phister and Mills, 2003; Delaherche et al, 2004; Tofalo et al, 2012; Willenburg and Divol, 2012; Vendrame et al, 2014), Hanseniaspora (Hierro et al, 2007; Phister et al, 2007), Saccharomyces (Martorell et al, 2005b; Hierro et al, 2007; Salinas et al, 2009), and Zygosaccharomyces (Rawsthorne and Phister, 2006) in wine and other fermentation processes. The main disadvantage other than cost and personnel training lies in the method’s inability to differentiate viable and non-viable microbes (Ivey and Phister, 2011).…”

Section: Methods To Detect Genetic Polymorphismmentioning

confidence: 99%

Genetic Polymorphism in Wine Yeasts: Mechanisms and Methods for Its Detection

Guillamón

Barrio

2017

Front. Microbiol.

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The processes of yeast selection for using as wine fermentation starters have revealed a great phenotypic diversity both at interspecific and intraspecific level, which is explained by a corresponding genetic variation among different yeast isolates. Thus, the mechanisms involved in promoting these genetic changes are the main engine generating yeast biodiversity. Currently, an important task to understand biodiversity, population structure and evolutionary history of wine yeasts is the study of the molecular mechanisms involved in yeast adaptation to wine fermentation, and on remodeling the genomic features of wine yeast, unconsciously selected since the advent of winemaking. Moreover, the availability of rapid and simple molecular techniques that show genetic polymorphisms at species and strain levels have enabled the study of yeast diversity during wine fermentation. This review will summarize the mechanisms involved in generating genetic polymorphisms in yeasts, the molecular methods used to unveil genetic variation, and the utility of these polymorphisms to differentiate strains, populations, and species in order to infer the evolutionary history and the adaptive evolution of wine yeasts, and to identify their influence on their biotechnological and sensorial properties.

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Section: Methods To Detect Genetic Polymorphismmentioning

confidence: 99%

Genetic Polymorphism in Wine Yeasts: Mechanisms and Methods for Its Detection

Guillamón

Barrio

2017

Front. Microbiol.

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“…This latter result is consistent with those of previous studies using the same primer set and qPCR procedure to detect and quantify B . bruxellensis in wine, where a limit of detection as low as 1 (Phister & Mills, ) and 10 (Tofalo et al., ) cells per reaction were observed.…”

Section: Resultsmentioning

confidence: 98%

“…Following the procedure described by Tofalo et al. (), the standard curve was created by inoculating Brettanomyces cells directly in red wine instead of water to avoid possible differences in the amplification efficiency of the Albanian wine samples and standard dilutions due to red wine PCR inhibitors, such as polyphenols, polysaccharides or tannins. In addition, a good correspondence was observed in the number of B .…”

Section: Resultsmentioning

confidence: 99%

“…The cultivable fraction of the B. bruxellensis population potentially occurring in the 22 wine samples was enumerated by the plate count method on Dekkera/Brettanomyces differential media (DBDM) (Rodrigues, Gonçalves, Pereira‐da‐Silva, Malfeito‐Ferreira, & Loureiro, ) as described by Tofalo, Schirone, Corsetti, and Suzzi (). Ten‐fold serial dilutions of the wine samples were prepared in sterile peptone water (peptone 1 g/L); 100 μL of each undiluted wine sample and serial dilutions were plated in duplicate and incubated at 25 °C for 10 days.…”

Section: Methodsmentioning

confidence: 99%

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Brettanomyces Spoilage in Albanian Wines Assessed by Culture‐Dependent and Culture‐Independent Methods

Lloha

Peçuli

Basha

et al. 2019

Journal of Food Science

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In the Albanian winemaking industry, there is little awareness of the potential detrimental effect of Brettanomyces in wines. The aim of this study was to detect and quantify Brettanomyces cells in 22 Albanian bottled wines, representing all the viticultural areas of Albania. A combined approach, including culture‐dependent (viable plate counting) and culture‐independent (qPCR) methods, was applied. Spoilage indicators (ethylphenols and total and volatile acidity), as well as the primary factors known to influence the growth of Brettanomyces in wine (pH, SO2, and ethanol concentration), were also investigated. Brettanomyces was detected in only five (one Merlot, four Sheshi i Zi) out of 22 samples analyzed using viable counting, with loads ranging from 1.30 ± 0.03 log CFU/mL to 3.99 ± 0.00 log CFU/mL, whereas it was never detected in the Kallmet samples. When qPCR was applied, Brettanomyces cells were detected and quantified in all of the samples with a generally low load ranging from 0.47 ± 0.13 to 3.99 ± 0.01 log cells/mL. As a general trend, the loads of spoilage by this yeast were low (≤1.92 log cells/mL), with the exception of five samples that were also positive by plate counting. A positive correlation between the growth of this spoilage yeast on Dekkera/Brettanomyces differential media and its detection at high levels by qPCR was observed. A significant positive correlation between Brettanomyces and the concentration of ethylphenols and volatile acidity was also found. In summary, the results of this study demonstrated the low incidence of Brettanomyces spoilage yeasts in Albanian red wines. Practical Application The awareness of Brettanomyces spoilage in the Albanian winemaking industry is very low. This study represents the first contribution to understand the extent of this spoilage yeast in Albanian autochthonous cultivars, which tend to have high economic value, to ensure product quality and safety. qPCR is confirmed to be a very sensitive method to rapidly detect Brettanomyces spoilage in wine samples.

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“…Arcore, Italy), according to the manufacturer's instructions but with some modifications as reported in Tofalo et al . ().…”

Section: Methodsmentioning

confidence: 97%

Characterization of specialized flocculent yeasts to improve sparkling wine fermentation

et al. 2016

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Detection of Brettanomyces spp. in Red Wines Using Real‐Time PCR

Abstract: Brettanomyces cells were detected using a qPCR method, optimized in this study, which allows to obtain results quickly.

Cited by 32 publications

References 41 publications

Genetic Polymorphism in Wine Yeasts: Mechanisms and Methods for Its Detection

Genetic Polymorphism in Wine Yeasts: Mechanisms and Methods for Its Detection

Brettanomyces Spoilage in Albanian Wines Assessed by Culture‐Dependent and Culture‐Independent Methods

Characterization of specialized flocculent yeasts to improve sparkling wine fermentation

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