Use of propidium monoazide for the enumeration of viable Brettanomyces bruxellensis in wine and beer by quantitative PCR

Vendrame, Marco; Manzano, Marisa; Comi, Giuseppe; Bertrand, J. E.; Iacumin, Lucilla

doi:10.1016/j.fm.2014.03.010

Cited by 24 publications

(19 citation statements)

References 55 publications

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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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“…Our findings are in agreement with those of Andorra et al (2010) who found that 6 μM is the optimum PMA concentration and that higher concentration of PMA will begin to inhibit the DNA amplification of the viable cells. In contrast, Vendrame et al (2014) were able to quantify a target spoilage yeast species by using 100 μM PMA, which is over 16 times greater than our optimum concentration. The discrepancy in optimum concentration between the two studies may be due to the difference in total cell density (~10 8 CFU/mL), length of PMA incubations, incubation temperature, light exposure time (Nkuipou-Kenfack et al, 2013), photo-crosslinking equipment, and the organisms used.…”

Section: Yeastmentioning

confidence: 64%

“…The discrepancy in optimum concentration between the two studies may be due to the difference in total cell density (~10 8 CFU/mL), length of PMA incubations, incubation temperature, light exposure time (Nkuipou-Kenfack et al, 2013), photo-crosslinking equipment, and the organisms used. Vendrame et al (2014) used PMA-qPCR to determine the quantity of the live yeast Brettanomyces bruxellensis in a wide range of densities (10 1 -10 8 CFU/mL) when mixed with the same species of dead yeasts at the PMA concentration of 100 μM. But this experiment was following only one species via the qPCR enumeration method.…”

Section: Yeastmentioning

confidence: 99%

“…Prior to the development of this method, the sole detection of live cells was accomplished by using reverse transcription (RT) to obtain cDNA, which was sequenced and subsequently quantified using qPCR. The use of this method was time consuming and was prone to error due to the many steps involved, the low stability of RNA under harsh conditions (Ivey and Phister, 2011), and the high variability in RNA transcription (Vendrame et al, 2014). The PMA method has been optimized for qPCR with respect to temperature, PMA concentration, and time of incubation (Andorra et al 2010;Nkuipou-Kenfack et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

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The use of propidium monoazide in conjunction with qPCR and Illumina sequencing to identify and quantify live yeasts and bacteria

Tantikachornkiat

Sakakibara

Neuner

et al. 2016

International Journal of Food Microbiology

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“…To overcome this problem, some authors have recently suggested the combined use of propidium monoazide (PMA) treatment with qPCR as a solution to successfully enumerate the B . bruxellensis viable cells in wine and beer (Vendrame et al., ).…”

Section: Resultsmentioning

confidence: 99%

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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Use of propidium monoazide for the enumeration of viable Brettanomyces bruxellensis in wine and beer by quantitative PCR

Cited by 24 publications

References 55 publications

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

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

The use of propidium monoazide in conjunction with qPCR and Illumina sequencing to identify and quantify live yeasts and bacteria

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

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