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Spoilage Microorganisms: Yeasts

Elizabeth Crawford
Dept. of Food Analysis & Nutrition
Institute of Chemical Technology, Prague, Czech Republic

Industrial and Food Microbiology Course


1. April 2014

General Considerations for Yeasts

Foods are considered habitats for


microorganisms
Most susceptible foods/beverages for yeast
spoilage have:
Low pH (5.0 or lower) which restricts the growth
of competing bacteria
High sugar & organic acid content (easily
metabolized carbon sources)

Compared with bacteria and molds, yeasts


play a minor role in food spoilage
Handbook of Food Spoilage Yeasts, Second Edition Tibor Deak (CRC Press 2007;
Print ISBN: 978-1-4200-4493-5; eBook ISBN: 978-1-4200-4494-2)

Growth Requirements for Yeasts

What defines a Spoilage Yeast?

Yeasts responsible for undesirable changes to


the sensory quality of the food.
Unwanted changes in the flavor, aroma and taste
of the final products.

In fermented alcoholic beverages, any yeast


changing the sensorial characteristics can be
regarded as a spoilage yeast.

Food technologists define asyeasts that spoil


a food product despite following GMP
standards.
* Loureiro V., Malfeito-Ferreira M. (Review Paper) Spoilage yeasts in the wine industry.
Int. J. of Food Microbiology 2003(86) 23-50.

Commodities Susceptible to Yeast Spoilage

Fresh and processed fruits


Fruit juices and soft drinks
Vegetables
Fermented alcoholic beverages
Beer and wines
Diary products
Milk, cheeses and fermented milk

Major Spoilage Yeasts in Foods & Beverages

Loureiro V., Malfeito-Ferreira M. (Review Paper) Spoilage yeasts in the wine industry.
Int. J. of Food Microbiology 2003(86) 23-50.

(Fresh) Fruits

Processed Fruits

High moisture content

Yeast associations are

(high aW)
pH range of 3-5
High concentration of
soluable carbohydrates

Overall a very nice


source for yeast growth

directly reflective of
harvesting and handling
practices
Contamination during
growing season, injuries
during harvesting and
handling

Yeasts in Fruits

Fruit Juices

Soft Drinks

Low pH
Cause of spoilage not
often from the
Low N2 and O2 content

ingredients, but most


often originates from the
environment for most
microorganisms, but
manufacturing process
amenable for yeast
Critical points of
growth
contamination are:
Fruit juices are higher in pumps, holding tanks,
nitrogenous compounds bottle washers and
& vitamins than soft
bottling lines
drinks, therefore they are
more susceptible to
yeast spoilage
Generally an adverse

Yeasts in Fruit Juices & Soft Drinks

Preventative Steps Against Yeast Spoilage

Chemical preservation
Addition of sulfur dioxide, sorbic acid, benzoic
acid, acetic acid

Pasteurization
Freezing
Concentration (lowering of aW)
Irradiation

Vegetables
Increased incidence of yeast spoilage in these
commodities due to storage/packaging in plastics,
minimal processing and stronger consumer demands
for ready-to-eat vegetables.
Spoilage is caused most frequently by Saccharomyces
cerevisiae.
Tomatoes are exceptional, in that yeasts represented
nearly 17% of fungal isolates from ripe, damaged and
decayed tomatoes.
Ready-to-eat vegetable salads were P. fermentans, P.
membranifaciens and unidentified Candida spp. and
Trichosporon spp.

Beer
Wild Yeasts - unwanted yeasts that enter into the beer
during fermentation
Two kinds - Saccharomyces and non-Saccharomyces genera
Origin: Brewery environment and pitching yeast

Phenolic off-flavor - coming from some strains of S.


cerevisiae wild yeasts that contain an enzyme that
decarboxyates wort phenolic acids

Strains (Pichia and Candida species) producing zymocins


(killer toxins) could completely eliminate pitching yeasts
causing fermentation to end

Film forming: responsible yeasts P. membranifaciens, P.


fluxuum and P. anomala.

Yeasts in Beer

Wine
Spoilage yeasts originating from the grapes are the
primary source of Dek. (Bret.) bruxellensis leading to
phenolic off-flavors

High concentrations of acetaldehyde can be achieved by


several Candida species and Scodes ludwigii, P. anomala,
and other Pichia species

Hanseniaspora (Kloeckera) species are also responsible


for high levels of acetic acid and its esters.

Common spoilage yeasts in bottled wine include Zygo.


bailii, S. cerevisiae, C. rugosa, P. membranifaciens, and C.
vini.

Preventing Wine Spoilage:


Rapid Screen & Quantification of Off-flavor
Phenolics using Ambient Ionization coupled with
High Resolution MS/MS
Elizabeth Crawford1, Paola Domizio2,3,
Brian Musselman1, C. M. Lucy Joseph2, Linda F. Bisson2,
Bart C. Weimer4 and Richard Jeannotte4,5
1IonSense,

Inc, Saugus, MA, USA


of Viticulture & Enology, Univ. of California-Davis, Davis, CA, USA
3Dipart. di Gestione Sistemi Agrari, Alimentari e Forestali (GESAAF), Univ. degli Studi di Firenze, Italy
4Dept. of Health & Reproduction, School of Veterinary Med, Univ. of California-Davis
5
Facultad de Ciencias, Univ. de Tarapac, Arica, Chile
2Dept.

47. Jahrestagung der Deutschen Gesellschaft fr Massenspektrometrie (DGMS)


03. Mrz 2014 Frankfurt am Main, Deutschland

Brettanomyces
Background:
Budding yeast found widely
distributed in nature
Discovered in beer in 1904
(Claussen), in wine
(Krumbholz &
Tauschanoff,1930) and again
in 1940 (Custers)

Produces a wide array of


aromatic compounds
Wine cellar contamination
was widespread
Brett characters can
compete with varietal
characters for dominance of
wine profile

Brett Wheel http://heysmartbeerdude.files.wordpress.com/2013/04/brett-aroma-wheel.jpeg (Access: 10 June 2013)

When Is It Spoilage?

High concentration, dominating wine profile


Conflict with wine matrix characters
Suppression of varietal character
Enhancement of off-notes
Lactic acid bacteria often found in wines with
Brettanomyces

Recovery Thresholds:
Chatonnet* has defined spoilage as:

>426 ppb of 4-ethylphenol (4-EP) and 4-ethylguaiacol (4-EG)


>620 ppb of 4-EP
50% of tasters can detect 605 ppb in wine or 440 ppb in water
of 4-EP
* Chatonnet, P.; Boidron, J. N.; Dubourdieu, D. Influence des conditions d levage et de sulfitage des vins rouges en
barriques sur leur teneur en acide actique et en thyl-phenols. J. Int. Sci. Vigne Vin. 2003, 27, 277-298.

Slide courtesy of Dr. L. Bisson, Dept. of Viticulture & Enology, Univ. of California-Davis

(GC-MS)

Slide courtesy of Prof. Jana Hajlov, ICT Prague, Czech Republic

DART MS/MS Method: Figures of Merit

Low

High

1
2
3
4
Mean
%CV

50
47.7
58.7
52.1
41.0
49.9
14.9

500
492.2
509.2
N/A
N/A
500.7
2.4

%Bias

-0.3

0.1

4-EP Conc.
(g/L)

Compare: Calculated Levels of 4-EP & 4-EG


Wine
Sample

DART HRAM
MS/MS

GC MS

4-EP
(g/L)

4-EG
(g/L)

4-EP
(g/L)

4-EG
(g/L)

Sample 04

854 *

197

845

203

Sample 05

518

157

563

161

Sample 06

52

ND

129

14

Sample 09

ND

ND

110

13

Sample 14

2774 *

492

2534

433

Yellow = Brettanomyces

* Levels above selected calibration range