PMID- 21925033
OWN - NLM
STAT- MEDLINE
DCOM- 20111222
LR  - 20191210
IS  - 1095-9998 (Electronic)
IS  - 0740-0020 (Linking)
VI  - 28
IP  - 8
DP  - 2011 Dec
TI  - Analysis and direct quantification of Saccharomyces cerevisiae and Hanseniaspora 
      guilliermondii populations during alcoholic fermentation by fluorescence in situ 
      hybridization, flow cytometry and quantitative PCR.
PG  - 1483-91
LID - 10.1016/j.fm.2011.08.009 [doi]
AB  - Traditionally, it was assumed that non-Saccharomyces (NS) yeasts could only 
      survive in the early stages of alcoholic fermentations. However, recent studies 
      applying culture-independent methods have shown that NS populations persist 
      throughout the fermentation process. The aim of the present work was to analyze 
      and quantify Saccharomyces cerevisiae (Sc) and Hanseniaspora guilliermondii (Hg) 
      populations during alcoholic fermentations by plating and culture-independent 
      methods, such as fluorescence in situ hybridization (FISH) and quantitative PCR 
      (QPCR). Species-specific FISH probes labeled with fluorescein (FITC) were used to 
      directly hybridize Sc and Hg cells from single and mixed cultures that were 
      enumerated by epifluorescence microscopy and flow cytometry. Static and agitated 
      fermentations were performed in synthetic grape juice and cell density as well as 
      sugar consumption and ethanol production were determined throughout 
      fermentations. Cell density values obtained by FISH and QPCR revealed the 
      presence of high populations (10(7)-10(8) cells/ml) of Sc and Hg throughout 
      fermentations. Plate counts of both species did not show significant differences 
      with culture-independent results in pure cultures. However, during mixed 
      fermentations Hg lost its culturability after 4-6 days, while Sc remained 
      culturable (about 10(8) cells/ml) throughout the entire fermentation (up to 10 
      days). The rRNA content of cells during mixed fermentations was also analyzed by 
      flow cytometry in combination with FISH probes. The fluorescence intensity 
      conferred by the species-specific FISH probes was considerably lower for Hg than 
      for Sc. Moreover, the rRNA content of Hg cells, conversely to Sc cells, remained 
      almost unchanged after boiling, which showed that rRNA stability is 
      species-dependent.
CI  - Copyright (c) 2011 Elsevier Ltd. All rights reserved.
FAU - Andorra, Imma
AU  - Andorra I
AD  - LNEG, Unidade Bioenergia, Estrada do Paco do Lumiar 22, 1649-038 Lisboa, 
      Portugal; Universidad Rovira & Virgili, Dept Bioquim & Biotecnol, Fac Enologia, 
      Tarragona 43007, Spain.
FAU - Monteiro, Margarida
AU  - Monteiro M
FAU - Esteve-Zarzoso, Braulio
AU  - Esteve-Zarzoso B
FAU - Albergaria, Helena
AU  - Albergaria H
FAU - Mas, Albert
AU  - Mas A
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20110812
PL  - England
TA  - Food Microbiol
JT  - Food microbiology
JID - 8601127
RN  - 3K9958V90M (Ethanol)
SB  - IM
MH  - Ethanol/*metabolism
MH  - Fermentation
MH  - Flow Cytometry/*methods
MH  - Hanseniaspora/genetics/*growth & development/metabolism
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Polymerase Chain Reaction/*methods
MH  - Saccharomyces cerevisiae/genetics/*growth & development/metabolism
MH  - Vitis/metabolism/microbiology
MH  - Wine/microbiology
EDAT- 2011/09/20 06:00
MHDA- 2011/12/23 06:00
CRDT- 2011/09/20 06:00
PHST- 2010/11/30 00:00 [received]
PHST- 2011/07/29 00:00 [revised]
PHST- 2011/08/07 00:00 [accepted]
PHST- 2011/09/20 06:00 [entrez]
PHST- 2011/09/20 06:00 [pubmed]
PHST- 2011/12/23 06:00 [medline]
AID - S0740-0020(11)00194-8 [pii]
AID - 10.1016/j.fm.2011.08.009 [doi]
PST - ppublish
SO  - Food Microbiol. 2011 Dec;28(8):1483-91. doi: 10.1016/j.fm.2011.08.009. Epub 2011 
      Aug 12.