YEASTS IN MILK AND DAIRY PRODUCTS 2761
Yak see Dairy Animals: Yak.
YEASTS IN MILK AND DAIRY PRODUCTS
H Seiler, Research Centre for Milk and Food,
Technical University of Munich, Germany
Copyright 2002, Elsevier Science Ltd. All Rights Reserved
Introduction
In organic matter that contains carbohydrates yeasts
can grow within a broad range of pH; thus they play
an important role in foodstuffs. In most instances,
growth of yeasts is undesirable in milk and dairy
products, because these microorganisms harbour a
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high risk of spoilage. In fermented dairy products
with low pH the frequently occurring organisms of
the bacterial families Bacillaceae, Enterobacteriaceae
and Pseudomonadaceae cannot proliferate, and there-
fore these are not responsible for the spoilage of such
foods. Acid dairy products with the addition of fruit
mixes, honey, cereal preparations, chocolate, etc. are
at most risk. These foods contain, in addition to
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lactose and small amounts of galactose, considerable
amounts of fructose and sucrose, therefore all yeast
species and not only the lactose-utilizing ones ®nd in
them excellent conditions for growth and fermenta-
tion, and consequently for the spoilage of the product.
On the other hand, yeasts are used in manufacture
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of some dairy products. This mostly pertains to
surface-ripened soft cheeses and semi-hard cheeses. In
traditional fermented milk products such as ke®r and
kumys, yeasts are used for the generation of speci®c
aroma ingredients, e.g. ethanol and carbon dioxide.
Yeasts are also used for some technological processes.
Nowadays, bakers' yeast, alcohols and biomass can
be produced from sweet whey.
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Raw and Market Milk
During milking, yeasts are introduced into the
milk mostly from above the ¯oors, litter, feed and
air, less frequently from a mastitic udder or the
milking machine. The total yeast count in raw milk
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is negligible at 101±103 cfu mlÿ1. The species most
often found are Candida intermedia, Can. para-
psilosis, Cryptococcus curvatus, Debaryomyces
hansenii, Galactomyces geotrichum, Issatchenkia
orientalis, Kluyveromyces marxianus, K. lactis (both
sometimes referred to as K. marxianus subsp.
marxianus, K. marxianus subsp. lactis, K. marx-
ianus subsp. bulgaricus, Can. kefyr), Pichia farinosa,
Pic. fermentans, Pic. membranaefaciens, Pic. anom-
ala, Trichosporon beigelii and Yarrowia lipolytica
(Table 1). There is a signi®cant relationship be-
tween udder health, and general hygiene during
milking and the yeast count in raw milk (see Liquid
Milk Products: Pasteurized Milk. Pasteurization
of Liquid Milk Products: Principles, Public Health
Aspects).
Even after homogenization at 55±65  C and heat
treatment at pasteurization conditions, milk can be
recontaminated with yeast and can develop mouldi-
ness, mustiness, and yeast odour. In spoilt milk,
amino acids and peptides can stimulate the growth
of starter cultures. During fermentation, this can
lead to overacidi®cation or to an undesirable
population displacement. Possibly, inhibitory sub-
stances would also be produced. The cultured
products can have defects such as `soft curd',
`separating and stripping', `¯avour defect' or
`lacking aroma'.
During pasteurization of milk, vegetative cells of
yeasts, blastospores, chlamydospores of Can. albicans
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2762 YEASTS IN MILK AND DAIRY PRODUCTS

Table 1 Yeast species frequently isolated from milk and dairy products

Species a Habitat/lactose utilization b,c

Bullera variabilis de

m
Candida atmosphaerica de
Candida auringiensis -/Laf
Candida boidinii f, y
Candida butyri b
Candida catenulata c, de, m, mm
Candida diddensiae b, cb, de

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Candida etchellsii cb, y
Candida ethanolica de, f, y
Candida fennica -/Laf
Candida glabrata f, y
Candida glaebosa m
Candida haemulonii f, y
Candida halophila -/Laf
Candida intermedia c, cb, cm, de, f, y/Laf
Candida lactis-condensi c, cm, y
Candida magnoliae f, y
Candida mannitofaciens -/Laf

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Candida parapsilosis b, c, cb, de, f, mm, y
Candida peltata mm
Candida rugosa b, c, cb, de, ds, k, m
Candida sake b, c, de, f, m, y
Candida santamariae f, de
Candida savonica f, de
Candida sorbophila de
Candida tenuis cb, k, mm/Laf
Candida tropicalis
Candida versatilis
ott b, nb, c, cb, f, k, m, y
c, cb, de, k, m/Laf
Candida vini b, c, ds
Candida zeylanoides c, cb, ds
Citeromyces matritensis (Candida globosa) cm, f, m
Clavispora lusitaniae (Candida lusitaniae) c, cb, de, f, mm, y
Cryptococcus albidus/curvatus/humicolus/laurentiae c, cb, de, f, m, y/Laf
Debaryomyces hansenii (Candida famata) b, nb, c, cb, de, f, k, m, r, y/Laf
Dekkera anomala (Brettanomyces anomalus) c, k/Laf
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Dekkera bruxellensis/custersiana (Brettanomyces spp.) k, m, y

Dipodascus capitatus (Geotrichum capitatum) c, mm
Filobasidiella neoformans (Cryptococcus neoformans) mm
Filobasidium ¯oriforme de
Galactomyces geotrichum (Geotrichum candidum) nb, c, cb, de, k, m, y
Geotrichum capitatum/fragrans/klebahnii nb, c, cb, de, m, y
Hanseniaspora uvarum (Kloeckera apiculata) de, f, m, y
Hanseniaspora vineae (Kloeckera africana) de, f, m, y
Hyphopichia burtonii (Candida chodati) c, m, y
Issatchenkia occidentalis (Candida sorbosa) nb, c, cb, de, f, k, m, y
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Issatchenkia orientalis (Candida krusei) nb, c, cb, de, f, k, m, y

Kloeckera lindneri f, y
Kluyveromyces cellobiovorus -/Laf
Kluyveromyces lactis (Candida sphaerica) b, nb, c, cb, de, f, k, m, y/Laf
Kluyveromyces marxianus (Candida kefyr) b, nb, c, cb, de, f, k, m, y/Laf
Leucosporidium scottii de
Metschnikowia pulcherrima (Candida pulcherrima) f, y
Metschnikowia bicuspidata/reukauf®i (Candida spp.) f, y
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Pichia angusta c, f, m, mm, y

Pichia anomala (Candida pelliculosa) nb, c, cm, de, f, m, mm, y
Pichia cactophila b, f, y
Pichia etchellsii b
Pichia fabianii (Candida fabianii) f, de
Pichia farinosa m, de
Pichia fermentans (Candida lambica) nb, c, k, m, y
Pichia guilliermondii (Candida guilliermondii) b, nb, c, de, f, y

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 YEASTS IN MILK AND DAIRY PRODUCTS 2763

Table 1 Continued

Species a Habitat/lactose utilization b,c

Pichia jadinii (Candida utilis) nb, c, cb, f, mm

m
Pichia kluyveri (Candida eremophila) nb
Pichia membranaefaciens (Candida valida) c, f, k, y
Pichia norvegensis (Candida norvegensis) b, de, y
Pichia pini f, y
Pichia pseudocactophila c, de, y
Pichia sorbitophila -/Laf

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Pichia triangularis (Candida polymorpha) c, cb
Rhodotorula glutinis/graminis/minuta/mucilaginosa b, c, cb, de, f, m, mm, y/Laf
Saccharomyces cerevisiae (Candida robusta) nb, c, cb, de, f, k, m, y
Saccharomyces dairensis/kluyveri nb, k
Saccharomyces exiguus (Candida holmii) nb, k, y
Saccharomyces servazzii k
Saccharomyces unisporus c, k
Saccharomyces turicensis k
Saccharomycodes ludwigii y
Sporobolomyces roseus/salmonicolor f, de
Sterigmatomyces halophilus cb

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Torulaspora delbrueckii (Candida colliculosa) nb, c, cb, de, f, k, m, y
Trichosporon beigelii/cutaneum c, cb, de, ds, m, y
Trichosporon asteroides/ovoides/pullulans b, nb, c
Williopsis californica/saturnus c, y
Yarrowia lipolytica (Candida lipolytica) b, c, cb, de, k, y
Zygoascus hellenicus (Candida hellenica) mm/Laf
Zygosaccharomyces rouxii c, f, y
Zygosaccharomyces bailii/bisporus c, f, y
Zygosaccharomyces ¯orentinus/mellis
ott k
a
In parenthesis, imperfect state.
b
Often found in: b, butter; nb, natural buttermilk; c, cheese; cb, cheese brine; cm, condensed milk; de, dairy
environment; ds, dairy sewage; f, fruit mix for fruit supplemented yoghurt and quark; k, ke®r; m, milk; mm, mastitis
milk; r, rennet; y, yoghurt; Laf, lactose assimilation and fermentation.
c
Bold print, very frequently found.
Data compiled from Barnett JA, Payne RW and Yarrow D (1990) Yeasts: Characteristics and Identi®cation.
Cambridge: Cambridge University Press; and other sources.
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and other species, teliospores, ballistospores, basidio-
spores, endospores, ascospores of ascosporogenous
yeasts, and arthrospores of the Endomycetaceae
(e.g. Gal. geotrichum) are destroyed. For the most heat-
resistant yeast, Zygosaccharomyces bailii, the follow-
ing inactivation values were determined: vegetative
cells: D1min ˆ 56  C, z ˆ 4  C; ascospores: D1min-
ˆ 64 C, z ˆ 3 C (see Sterilization of Milk and Other
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Gram-negative organisms. These bacteria, and not
yeasts, are responsible for the shelf-life limit of 7±14
days of pasteurized milk.
Various milk products are heat-treated at
90±110  C, for a short time. Under such conditions,
all bacteria as well as the ascospores from thermo-
philic moulds are destroyed, but not the spores of
bacilli and clostridia. These can cause microbio-

Products). Under low water activity (aw) conditions,
for example when yeast cells are pressed in porous
gaskets, the inactivation from heating may be insuf-
®cient. Areas in the processing plant that pose a risk
of recontamination are heat exchangers, cooling
water, ®lling equipment, air and packaging materials.
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logical problems. Yeasts can only contribute to or
cause spoilage in case of recontamination. Under
high-temperature pasteurization (e.g. 125  C for
5 s) and sterilization (e.g. 109±115  C for 20±40
min) conditions, all microorganisms except for
some heat-resistant bacterial endospores are in-

Generally, in pasteurized milk and pH-neutral
dairy products recontamination with yeasts is of little
importance to spoilage. Acinetobacter spp. and many
Gram-positive bacteria including spores of Bacillus
spp. and Clostridium spp. survive pasteurization.
Moreover, because the ®lling operation is not sterile,
the milk is most frequently recontaminated with
activated. Nevertheless, it has been observed that
the occurrence of spoilt milk is connected with
the presence of heat-stable proteases and lipases,
which originate from high numbers of micro-
organisms present in milk before heating (see
Microorganisms Associated with Milk). However,
these enzymes come from Pseudomonas strains,

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2764 YEASTS IN MILK AND DAIRY PRODUCTS
not from yeasts. Enzymes produced by yeasts
are completely inactivated during these heat
treatments.
Butter and Natural Buttermilk
Butter made in a dairy plant and stored at a low
temperature seldom undergoes microbial spoilage
within its shelf-life. The ®ne dispersion of water, the
low pH value (in cultured butter) and the low solu-
bility of oxygen in fat hinder the growth of micro-
organisms. The product surface is exposed to a limited
hygienic risk. Yeasts can grow there and damage
the butter, for instance by lipolysis or discoloration.
Improper production conditions or unsterile storage
can facilitate spoilage caused by yeasts. Home-made
butter contains many serum droplets with lactose
as a convenient nutrient for microorganisms; its
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hygienic status leaves much to be desired. Here yeasts
can reach high counts. Addition of NaCl reduces the
risk of spoilage.
Natural buttermilk should at most contain
200 cfu mlÿ1 yeast cells. A large number of micro-
biological defects can be traced to yeasts, since they
can still grow at pH <3.5 and at 0  C. The spoilage
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is caused by proteolysis, lipolysis and carbohydrate
fermentation with the formation of gas. Overacidi-
®cation as well as consistency and sensory problems
can arise; packages can blow. The yeast species
most often found in industrially produced buttermilk
are Deb. hansenii, Can. tropicalis, Galactomyces
spp., Geotrichum spp., Issatchenkia spp., Kluyver-
omyces spp., Pic. anomala, Pic. kluyveri, Saccharo-
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myces cerevisiae and Torulaspora delbrueckii. In
buttermilk made from traditional Jordanian butter
Sac. cerevisiae, K. marxianus, Pic. kluyveri, Gal.
geotrichum, Iss. occidentalis, Ts. delbrueckii, Can.
tropicalis and Tsp. ovoides were the most prevalent
yeast species.
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Ice Cream and Frozen Yoghurt
Milk products marketed in the frozen state cannot be
spoiled by yeasts. The growth rate and metabolic
activity at temperatures <0  C are irrelevant. How-
ever, the ®nished product should not be more than
very minimally contaminated; the product cannot
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be permitted to thaw out in between.
A health risk from yeasts can then exist when
facultative pathogens, for example Can. albicans and
Can. dubliniensis, as well as Filobasidiella neofor-
mans subsp. neoformans, Fil. neoformans subsp.
bacillisporus and Fil. neoformans subsp. gattii,
appear in food or on food-contaminated equipment.
The opportunistic pathogenic yeast species Can.
parapsilosis, Can. tropicalis, Can. haemulonii,
Cryptococcus spp., Deb. hansenii, Iss. orientalis, Pic.
guilliermondii, Rhodotorula spp., Sporobolomyces
spp. and Tsp. beigelii among others, have been
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known to cause infections in susceptible individuals
such as infants, seniors, immune-compromised in-
dividuals, persons with AIDS, diabetics, alcoholics,
and pregnant women. In young children, oral thrush
and nappy rash are not unheard of; allergies can also
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be involved. Immune-suppressed individuals can
suffer from a serious mycosis of the organs. An attack
by Filobasidiella (teleomorph of Cryptococcus) leads
to the dreaded Cryptococcus mycosis of the brain,
lungs, bone marrow, kidneys, respiratory tract,
digestive tract, eyes, skin, central nervous system and
nails. However, these yeasts are ubiquitously dis-
tributed. The species Can. albicans and Fil. neofor-
mans, with the highest potential risks, are very
seldom found in milk and dairy products. Quite a few
of the other known species occur frequently in food,
but in contrast to the clinical isolates, they are not
very virulent and are not resistant to antibiotics.
In food, potentially dangerous concentrations of
toxins are not produced. Opportunistic pathogenic
yeasts are generally found in milk from cows with
mastitis.
Cultured Milk Products
Cultured milk products (fermented milk, sour cream,
yoghurt, drinking yoghurt, cottage cheese, cream
cheese, etc.) should be free of yeasts. These foods are
ideal media for the propagation of yeasts, since they
exhibit the low pH of 4±6, optimal for yeast growth.
The decimal doubling time in fruit yoghurt for Sac.
cerevisiae without shaking is D30 C ˆ 5 h, D20 C ˆ
10 h, D10 C ˆ 62 h, D4 C ˆ 84 h and for Gal.
geotrichum is D30 C ˆ 6 h, D20 C ˆ 12 h, D10 C ˆ
96 h, D4 C ˆ 7 days.
Due to the acidic environment (pH 3.8±4.5),
there is limited competition from bacteria in yoghurt.
Most of those that can still grow alongside the yeasts
and moulds are the lactic acid bacteria (LAB, i.e.
Lactobacillus, Lactococcus, Weissella and Leuco-
nostoc spp.), streptococci, enterococci, bi®dobacteria,
propionic acid bacteria, acetic acid bacteria, Micro-
coccus kristinae and Zymomonas mobilis. The
enterobacteria, for example, die off quickly in yoghurt.
Spores of bacilli and clostridia cannot germinate.
For the acid-tolerant anaerobic Pediococcus spp.,
Pectinatus cerevisiiphilus and Megasphaera cerevi-
siae, this environment contains too much oxygen.
Therefore, next to contamination with moulds,
contamination with yeasts is the largest microbial
problem in these types of products. The annual
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economic losses to the dairy industry are substantial.
Fruit-containing fermented milk products spoil
quickly, owing to the high fructose and sucrose con-
tent of the fruit preparations, which considerably
promote yeast growth and fermentation. A collapse
(with nonfermenting yeasts) or swelling (with fer-
menting yeasts) of the cups, a change in texture,
product discoloration, off-¯avours, off-tastes or
visible microbial colonies on the product surface are
the evidence of this spoilage.
The fruit preparations are delivered to the dairy in
large containers. As a rule, even negligible contam-
ination with yeasts in these containers can lead to
immense losses. An entire day's yoghurt production
can be affected. These products cannot be offered
for sale. In Germany, for example, such contam-
inated products are declared as hazardous waste
and consequently have to be discharged on a special
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waste dump. The risk of damage can be reduced
by ®lling temperature of the fruit mix of <15  C,
consistent chilled storage of the container, the
avoidance of a stepwise emptying of the container,
a high sugar concentration in the fruit preparation
and prompt processing. Large dairies increasingly
produce the fruit preparations themselves. This in-
creases the microbiological safety considerably, since
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the process of portioning and transport are elim-
inated. The fruit preparation is pumped directly
from the cooking boiler or tubular heat exchanger
through a cooler into the storage tank, from which
the portions for the fruit yoghurt are directly drawn.
In the preparation of heat-treated yoghurt, the
mixture of fruit preparation and cultured milk is
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heated to 70±80  C in a scraped-surface or tubular
heat exchanger (immediately before ®lling into the
cups). This type of yoghurt has a shelf-life of 1 to
5 months longer than yoghurt with viable LAB
cultures. This product generally cannot undergo
changes due to the activity of LAB and fruit mix
contaminants such as other acid-tolerant bacteria as
well as yeasts and moulds.
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With the exception of the `white mould' Galacto-
myces geotrichum (anamorphic state ˆ Geotrichum
candidum; formerly Oidium lactis or Oospora lactis),
there are no fungi found that are limited to only the
fruit preparations or to the yoghurt portion. The
xerophilic species Zygosaccharomyces spp., Citer-
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omyces matritensis, Can. versatilis, Pic. etchellsii, Pic.
ciferrii and Pic. sorbitophila are considerably more
common in fruit preparations than in pure milk
products. On the other hand, the typical cheese yeast
Deb. hansenii is found more often in milk than in the
fruit mix. The fermented milk portion is occasionally
sweetened with sucrose syrup. In such cases, it is
not possible to distinguish between yeast species
YEASTS IN MILK AND DAIRY PRODUCTS 2765
originating from the production environment of the
dairy and that of the fruit mix processor.
In the dairy plant environment (¯oors, walls,
equipment) the yeast species identi®ed most com-
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monly are Deb. hansenii, Clavispora lusitaniae,
Rhodotorula spp., Cryptococcus spp., Can. inter-
media, Can. parapsilosis, Can. sorbophila, K. marx-
ianus, Ya. lipolytica, Issatchenkia spp., Trichosporon
spp. and Gal. geotrichum. The dominant yeast
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species from fruit preparations and contaminated
fruit-containing cultured milk products are Sac. cer-
evisiae, Pic. anomala, Pic. fabianii, Pic. membranae-
faciens, Hs. vineae, Hs. uvarum, Deb. hansenii, Can.
parapsilosis, Can. tropicalis, Can. intermedia, Ts.
delbrueckii and Cs. lusitaniae. The moulds Mucor
spp. and Aureobasidium spp. as well as the un-
pigmented algae Prototheca spp. show yeastlike
growth at submerged cultivation, hence a risk exists
of mistaking them for yeasts. Moreover, Mucor spp.
and Prototheca spp. produce large quantities of
carbon dioxide and are opportunistic pathogens.
Prototheca spp. have a physiological reaction pattern
similar to that of Sac. cerevisiae. A yeast-killing toxin
designated mycocin HMK (Hansenula mrakii killer
strain) produced by Williopsis mrakii (reclassi®ed)
was reported to have potential applications in con-
trolling yoghurt spoilage caused by yeasts (see Fer-
mented Milks: Yoghurt Types and Manufacture).
pH-Neutral Fruit-Containing
Milk Products
In pH-neutral milk products, e.g. milk rice, semolina
pudding or milk pudding, with additions based on
fruit, cocoa, nuts, vanilla husks, vitamin mixes or
cereals, yeasts are (next to bacilli and moulds) the
most common spoilage organisms. All the yeast
species that can be found in the added preparations,
in the dairy-based component, and in the dairy
environment are possible contaminants. The spoilage
organisms are strictly oxidative as well as ferment-
ative yeasts. The usual appearances of spoilage are
blowing or sucking in of containers and a conspicuous
change in the product consistency or in ¯avour and
taste. In most cases of spoilage the fault lies with the
added preparations, owing to the favourable nutrient
conditions (glucose, fructose, sucrose, organic acids),
as well as to the occasional long storage time of
the product containers at 20  C; in such situations,
a considerable increase in yeast count can occur.
In addition, underlaid products are often produced,
which means that ®rst the fruit preparation is ®lled
in the cup and then the hot dairy portion at a tem-
perature of 70±80  C is added on top. No yeasts
appear in the milk portion. Yeasts are also seldom
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2766 YEASTS IN MILK AND DAIRY PRODUCTS
found in toppings made from whipping cream or
vegetable oil foams, since these have been heated and
there is little possibility for yeasts to multiply.
Kefir
Ke®r is a type of fermented milk produced by a mixed
¯ora consisting of yeasts, various lactic acid bacteria
and acetic acid bacteria (see Fermented Milks: Ke®r).
This ¯ora forms nodules known as `ke®r grains'.
Typical ke®r contains ethanol and predominantly
L(‡)-lactic acid and it has effervescence owing to the
presence of CO2. Under German regulations, ke®r
must contain at least 0.05% (w/v) ethanol and CO2
produced by the yeast fermentation. According to
the Swiss food manual, there should be a minimal
yeast count of 105 cfu gÿ1. The International Dairy
Federation standard proposes a minimal yeast count
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of 104 cfu gÿ1, but without being species-speci®c.
Kumys
Kumys (also kumiss, koumyss, koumiss, coomys,
kimiz) is a fermented beverage made from mares'
milk (see Fermented Milks: Asia). Its origins are in
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central Asia. The ¯ora of kumys includes yeasts and
various lactic acid bacteria. Owing to a relatively high
lactose concentration in mares' milk, alcohol content
in kumys can reach approx. 3.5%.
Other Fermented Products
Containing Yeasts
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A Japanese patent describes milk fermentation with
Bi®dobacterium longum together with yeasts under
not strictly anaerobic conditions. The growth of these
fastidious bacteria is positively in¯uenced by the
change in the milk environment brought about by the
yeast growth.
Taxonomically, the yeast-like fungi Galactomyces
spp., Geotrichum spp. and Dipodascus spp. are
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seen to be somewhere between yeasts and moulds.
Different authors will place the genera in one taxo-
nomic order or the other. In Finland, viili is a popular
set-curd milk product. It is consumed pure, or
sweetened, with jam and raisins, or with cereal.
The milk is inoculated with a mesophilic, aroma-
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producing LAB culture and with Gal. geotrichum.
Because of the growth of the strictly oxidative white
mould on the product/air interface, viili has a matte,
velvety, white to light yellowy surface. The Gal.
geotrichum works to develop the ¯avour and also
hinders autooxidation of fats as well as contamina-
tion of the product surface by wild moulds. The
strains used are only those that have minimal lipase
activity. Galactomyces geotrichum and similar
species can produce a nut-like ¯avour. Similar prod-
ucts are the Norwegian tettemelk and Swedish laÊng®l
(see Fermented Milks: Northern Europe. Geotrichum
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candidum).
Leben is a fermented milk product from the Arab
countries, similar to ke®r. This product is made from
fresh milk using a mesophilic LAB and thermophilic
yoghurt culture as well as yeasts. Correspondingly,
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one also ®nds ethanol, acetoin and diacetyl in leben.
Again K. marxianus becomes dominant within the
yeast species. Generally, however, the micro¯ora is
not homogeneous, because leben is mostly home-
made. Coliform bacteria, black and green moulds and
enterococci are also regularly found in this product
(see Fermented Milks: Middle East).
Similar yeast-containing products are yaghurt
(Middle East), dahi and misti dahi (India), acid-
ophilus yeast milk, busa, kuban, kurunga, prohlada
and salomat (Russia), rob (Egypt), omaere (Africa),
skyr (Iceland), samokisselis (Yugoslavia), airan and
arsa (Asia), aker (Tibet), airag, khoormog, tschigan
and umdaa (Mongolia), matzoon (Armenia), brano
milk (Bulgaria), felisowka (Poland), galazyme
(France), cellarmilk (Norway), hooslanka, urda and
zhentitsa (Carpathian Mountains) (see Fermented
Milks: Types and Standards of Identity).
Cheese and Brine
White Cheese
The yeast species most frequently isolated from
white cheeses (quark, Gervais, cottage cheese, cream
cheese) are Gal. geotrichum, K. marxianus, K. lactis,
Pic. membranaefaciens, Pic. guilliermondii, Deb.
hansenii, Tsp. beigelii, Iss. orientalis and Ya. lipo-
lytica. The yeast count thresholds for defects such as
slightly vs. strongly `yeasty, fermenting, fruity, old,
musty, bitter', are 104±105, and 105±106 cfu mlÿ1,
respectively. The effect on sensory pro®le depends
upon the species. The lowest count value to cause
a sensory change was shown by Gal. geotrichum,
followed by Kluyveromyces spp., Pic. membranae-
faciens, Sac. cerevisiae, Deb. hansenii, Iss. orientalis,
Ya. lipolytica and Sac. exiguus. A separator-curd
which had been initially contaminated with
100 cfu gÿ1, showed the ®rst signs of sensory
defects after 5±7 days at 10  C; after 10 days, it was
spoiled. Therefore, a good product should have
<100 cfu gÿ1 contaminants. The generation time of
yeasts in curd at 2, 4, 6 and 10  C is approximately
100, 50, 20 and 10 h, respectively. Yeasts have a direct
correlation with the quality of white cheese, and
their absence is an important indicator of good
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manufacturing practice (GMP). With the presence of
yeasts the shelf-life of products at 10±6  C is limited
to 10±15 days.
Soft, Semi-Hard and Hard Cheese
Positive aspects Yeasts are important in the devel-
opment of surface-ripened cheese (see Cheese: Smear-
Ripened Cheeses). It is commonly known that there
exists a synergistic relationship between yeasts, Brevi-
bacterium linens, Microbacterium lacticum, micro-
cocci and LAB. The survival of lactobacilli is
enhanced by the presence of yeasts. The low pH res-
ulting from the metabolism of the LAB is raised
through lactate utilization by yeasts as well as the
formation of alkaline products through proteolysis,
so that aerobic, acid-sensitive bacteria such as Brev.
linens and micrococci can develop. For the growth
ob
of this bacterial ¯ora, vitamins and amino acids are
provided by yeasts. At the same time, yeasts are a
protective culture against undesired enterobacteria,
Clostridium tyrobutyricum, Staphylococcus aureus
or wild moulds. Through the development of gas
in Gorgonzola and other cheeses, an open doughy
structure is promoted. Aromatic components are
formed through lipolysis and proteolysis. Some yeast
ott
species or single strains that hydrolyse speci®c
casein fractions, have a positive effect on the growth
of LAB and Penicillium roqueforti in blue-veined
cheeses. Aminopeptidases and carboxypeptidases
from Gal. geotrichum are essential contributors to
the breakdown of bitter peptides. Extracellular and
intracellular peptidases with varying pH optima are
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found, respectively, in Ya. lipolytica and Can. cate-
nulata, and in Tsp. beigelii and Deb. hansenii. The
concentration of soluble nitrogen increases; glutam-
ate and aspartate as well as tryptophan, leucine,
methionine, phenylalanine and other amino acids
are deaminated. Sensorially important alcohols and
aldehydes are produced by utilization of amino
acids and carbohydrates. The intracellular enzymes
w.
are present after yeast autolysis, that is, in the
late stages of ripening. In cheese ripening studies
done with aseptic curd slurries, Deb. hansenii pro-
duced an acidic, fruity and cheesy aroma. Other
cheese isolates did not reveal similar tendencies.
The species Ya. lipolytica, Tsp. beigelii and Gal.
ww
geotrichum had a positive in¯uence on the aroma
as a result of the breakdown of proteins and pep-
tides. The species Cs. lusitaniae, Pic. jadinii and
Williopsis californica showed weak proteolytic activ-
ity, but caused an increase in pH owing to
their ability to utilize lactate. The strongest lipo-
lytic and proteolytic activity was demonstrated by
Ya. lipolytica. Debaromyces hansenii and other yeast
YEASTS IN MILK AND DAIRY PRODUCTS 2767
species are able to assimilate cadaverine, histamine,
putrescine and tyramine.
Negative aspects Strong yeast growth on cheese
m
can lead to defects in aroma and ¯avour (yeasty,
mouldy, putrid, overripe, alcoholic, musty, fer-
mented, earthy, spicy, ammonia, pungent, rancid,
sweet and gassy). Sometimes, Pic. jadinii causes
blowing in young cheese. The rind can become
.co
slimy and even semi-¯uid owing to Cryptococcus
spp. activity, because in this genus an extracellular
starchlike capsule is produced. The yeast species
Pic. anomala reduced the growth of P. roqueforti.
Some DOPA-positive strains of Deb. hansenii and
Ya. lipolytica with high levels of metabolic activity
produce an undesirable brownish pigment. With
acid curd cheeses, an overdominance of Gal. geo-
trichum is an indicator of spoilage; Ya. lipolytica
was found to generate the strongest odour, from
overripe to putrid; while Tsp. beigelii was judged
to produce a rancid odour. In mixed cultures, Iss.
orientalis inhibited growth of Lactococcus lactis,
whereas Lc. lactis stimulated growth of Iss. orien-
talis. In pickled cheese brine, acid-consuming
yeasts increased the pH of the brine to a level
that enabled the development of St. aureus. This
indicates the need to monitor yeast contamination
in cheeses preserved by a combination of acid and
high salt content. In Brick cheese, a low temperature
during ripening, high salt content or drying of the
cheese surface suppresses yeast growth, thereby
prolonging the ripening process. Interactions among
the various LAB and yeast organisms take place in
the overall product formation. Because of this, one
must specify the yeast count, species and strains
favourable to the respective cheese type. In some
countries, the antifungal agents natamycin (pimar-
icin), propionate or sorbate are used to inhibit the
growth of yeasts on the surfaces of hard and semi-
hard cheeses.
Starter cultures Yeasts, together with conidia of
P. candidum, can be added as a starter culture
directly to the milk vat (for white surface-ripened
cheeses such as Camembert and Brie), or sprayed or
brushed onto the rind together with the red smear
¯ora (red cheese, acid curd cheese, Croute Mixte
cheese). Culture suppliers offer preparations with
Deb. hansenii and Pic. jadinii for the production of
Brick cheese. For manufacturing acid curd cheeses,
a preparation with `mycoderma' species is offered.
Mycoderma were previously known as the pellicle-
forming fungi Gal. geotrichum, Dipodascus capita-
tus, Geotrichum spp., Tsp. beigelii, Iss. orientalis,
Pic. membranaefaciens and Pic. fermentans. The
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2768 YEASTS IN MILK AND DAIRY PRODUCTS
cultures contain selected strains with a de®ned
ability to metabolize lactate, citrate, acetate (deacidi-
fying activity), as well as galactose and lactose
(growth rate). They should also have proteinase,
peptidase and esterase activity (neutralization, ¯a-
vour, taste), lipase and phospholipase activity
(aroma), be able to produce CO2 (doughy structure),
tolerate anaerobiosis, acid, cold and NaCl, have high
growth rate and have an effect on cheese surface
appearance as well as inhibit wild moulds (see
Cheese: Starter Cultures: Speci®c Properties. Geo-
trichum candidum).
Interior of cheeses In the interior of cheeses made
under strict standards of hygiene, the yeast count
is low, at 101±103 cfu gÿ1. Metabolic products of
importance are not expected in these cases. The
process of diffusion between the rind and the core
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brings about deacidi®cation. In the interior of raw
milk cheeses the counts and the diversity of the
¯ora are higher than in those made from pasteurized
milk. The species identi®ed inside the raw milk
cheese TeÃte de Moine were: Cs. lusitaniae, Deb.
hansenii, Pic. pseudocactophila, K. marxianus,
Rhodotorula mucilaginosa, Deb. capitatus and Pic.
ott
jadinii. Owing to the large differences in the yeast
counts of the surface, middle and core layers of
the cheese, distinct variations in the metabolic
products are observed. For example, in Taleggio
cheese after 35 days of ripening at 3±10  C, the
three layers mentioned above had pH of 6.5, 5.5
and 5.2, respectively. In cheeses which should have
an adequate yeast ¯ora in the interior to soften
alk
the cheese structure through CO2 production, the
yeasts have to be added to the milk vat. Here,
a naturally high yeast count can be established.
The speed of deacidi®cation of the core area will
depend upon the size of the cheese and its dry weight.
Yeast species and counts on cheese surface In the
®rst days of the ripening period the yeast count
w.
on the surface increases rapidly and after 8±10
days reaches a maximum of 106±109 cfu gÿ1 or
108 cfu cmÿ2. This count decreases slightly during
further ripening. The softer the rind, the higher
the initial yeast count, and therefore, the smaller
the decrease in viable count. On soft, semi-hard
ww
and hard cheeses, diverse yeast species are seldom
found, but various species of the ¯ora may dominate.
The predominant species are Deb. hansenii, Tsp.
beigelii, Ya. lipolytica, K. marxianus, Can. zeyla-
noides, Can. catenulata, Ts. delbrueckii and Gal.
geotrichum.
In well-ripened Mozzarella cheese, Sac. cerevisiae
and Kluyveromyces spp. were primarily found; in
cheeses with a yeasty ¯avour defect the species Ya.
lipolytica, Iss. orientalis and Can. parapsilosis
were found in addition to Sac. cerevisiae and Kluy-
veromyces spp. On surface-ripened, acid-coagulated
m
skim milk cheeses, Deb. hansenii and Pic. mem-
branaefaciens were the predominant yeast species.
On Brick cheese, the dominant yeasts were Deb.
hansenii and Gal. geotrichum. Blue-veined cheese
surfaces overwhelmingly had Deb. hansenii, and
.co
on the inside Deb. hansenii, K. marxianus, Sac. cer-
evisiae, Pic. anomala, Tsp. delbrueckii and Gal.
geotrichum. In Harzer cheese, a shift in the ¯ora
was observed. At the beginning, those primarily
found in dry quark were K. marxianus, Tsp. beigelii,
Iss. orientalis and Dekkera anomala, while in the
early stages of ripening Tsp. beigelii and Can. cate-
nulata were predominant. At the end, only Ya. lipo-
lytica could be identi®ed. Kluyveromyces marxianus,
K. lactis and Sac. cerevisiae contribute to the
characteristic open structure of Gorgonzola. In foil-
ripened Raclette model cheese, a combined yeast
starter culture of Deb. hansenii, Pic. jadinii, Ya. li-
polytica and Gal. geotrichum was found to be
advantageous.
In general, different population spectra are ob-
served depending on milk quality, water and salt
content of the cheese, production hygiene, possible
addition of yeast culture, storage temperature, stage
of ripening, competing ¯ora and location of cheese
sampling. Microbial ¯ora is also affected by the
geographic region, manufacturer and range of prod-
ucts made on site, production lot or batch, age of the
saline bath and season of the year, as well as the
methods of isolation, enumeration and identi®cation
of yeasts used.
As a rule, Deb. hansenii is the prevailing species
identi®ed. Its role in the deacidi®cation of cheeses
must be considered essential. This is due to its
favourable metabolic capacity in the speci®c envir-
onment (fermentation of glucose, galactose, etc.;
metabolism of lactose, lactate, citrate, etc.; proteo-
lysis), high rate of sodium expulsion, potassium
uptake potential and its high salt tolerance
(aw minimum 0.85). To protect against plasmo-
lysis caused by NaCl and dehydration, osmotolerant
yeasts produce large quantities of the polyols such
as glycerol, D-arabitol, xylitol, erythritol and manni-
tol. Debaromyces hansenii still shows growth to
0.3 OD after 100 hours incubation in a broth
medium containing yeast extract, malt extract,
glucose and 18% (w/v) NaCl. It is followed by K.
marxianus (15% NaCl), Ts. delbrueckii (14%),
Ya. lipolytica (14%), Pic. farinosa (14%), Can.
versatilis (14%), Sac. unisporus (14%), Can.
zeylanoides (13%), Can. catenulata (13%), Sac.
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cerevisiae (9%; aw minimum 0.94), Gal. geo-
trichum (2%) and Tsp. beigelii (2%). For Deb.
hansenii, the optimum for growth lies between 0%
and 11% NaCl, and the inhibitory concentration of
NaCl is 24%. Distinctly lower are the optimum
concentrations for Cs. lusitaniae (0±6%) and Sac.
cerevisiae (0±2%). The xerotolerant Zygosacchar-
omyces spp. and Citeromyces matritensis have a high
tolerance to low aw values (minimum 0.65 ÿ 0.60)
in an environment with high sugar concentrations,
yet are relatively sensitive to NaCl.
In values of osmotic tolerance reported in the
literature the speci®c test substance is generally
named. For example, for a strain of Zs. rouxii the
following aw values are given: glucose/fructose
0.71, ammonium sulphate 0.82, xylose 0.89,
KCl 0.87, PEG 400 (polyethylene glycol with an
average molecular weight of 400) 0.88, NaCl
ob
0.89, PEG 200 (polyethylene glycol with an average
molecular weight of 200) 0.95. As a rule, different
values are observed depending on whether strains
were cultivated on agar plates (colony forming
capability as the criterion) or in broth (OD as the
criterion), whether gas production was used as
a parameter (fermentation as the criterion), and
whether the ability to bud could be determined
ott
under a microscope (initiation of reproduction as
the criterion). Also of diagnostic importance is
what incubation temperature and time were used, as
well as which factor is limiting their ability to grow
(OD), ferment (amount of gas) or bud (number of
budding cells). These values therefore allow differ-
entiation between various strains of a species on the
alk
basis of differing test conditions and de®ned posit-
ive metabolic activities. Because of this, no general
values can be given. Strains of the osmotolerant to
osmophilic species Can. etchellsii, Sterigmatomyces
halophilus, Pic. triangularis and Can. haloni-
tratophila are more salt tolerant than Deb. hansenii.
These do not grow at low NaCl concentrations
(optimum is 11±13%) and they generally grow more
w.
slowly than Deb. hansenii.
Brine In brines for surface-ripened soft, semi-hard
and hard cheeses with 12% to 22% NaCl, the
dominant species are Deb. hansenii and Can. versa-
tilis. Also worth mentioning are the species Tsp.
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beigelii, Can. parapsilosis, Can. tropicalis, Can.
polymorpha, Can. zeylanoides, Can. rugosa, Can.
intermedia, Can. tenuis, K. marxianus, K. lactis, Cs.
lusitaniae, Iss. orientalis, Pic. jadinii, Geotrichum
spp., Ya. lipolytica, Saccharomyces spp. and Ts.
delbrueckii. Several other species are less frequently
isolated. In brine of Nabulsi cheese, a Jordanian tra-
ditional boiled white cheese usually made from
YEASTS IN MILK AND DAIRY PRODUCTS 2769
sheep's milk and kept in brine with 20% NaCl, the
halotolerant species Deb. hansenii and Can. para-
psilosis and the halophilic species Pic. triangularis,
Pic. etchellsii and Stm. halophilus were found. The
m
yeast counts were between 102 and 106 cfu mlÿ1. For
the determination of halophilic species, one needs an
isolation medium with salt (e.g. 15% NaCl). In brines
of Feta made in Germany and Italy the yeast species
found were Deb. hansenii, Pic. membranaefaciens,
.co
Ya. lipolytica, K. marxianus, Zs. bicuspidata, Can.
magnoliae, Can. zeylanoides, Sac. cerevisiae and Ts.
delbrueckii. The yeast counts were mostly between
105 and 107 cfu mlÿ1. Two products were free of
yeasts because they had been pasteurized. From the
consumer's point of view, packages should be used
up quickly after opening, so that yeasts do not mul-
tiply with the introduction of oxygen and develop
a musty, yeasty, soapy or rancid ¯avour in the
product.
See also: Cheese: Starter Cultures: Specific Properties;
Smear-Ripened Cheeses. Fermented Milks: Types and
Standards of Identity; Northern Europe; Middle East;
Asia; Kefir; Yoghurt Types and Manufacture.
Geotrichum candidum. Kluyveromyces spp. Liquid
Milk Products: Pasteurized Milk. Microorganisms
Associated with Milk. Pasteurization of Liquid Milk
Products: Principles, Public Health Aspects.
Further Reading
Engel G, Einhoff K, Prokopek D and Teuber M (1987)
Beziehungen zwischen Hefenart, Hefenkeimzahl und
hefenbedingten sensorischen Fehlern in Magerquark
nach Zugabe de®nierter Hefenarten. Milk Science
International 42: 428±430.
Fleet GH (1990) Yeasts in dairy products: a review.
Journal of Applied Bacteriology 68: 199±211.
Geiges O and Spillmann H (1994) Yeast ¯ora of
commercial ke®r and ke®r cultures. Proceedings of the
24th International Dairy Congress, Melbourne, p. 403.
Jakobsen M and Narvhus J (1996) Yeasts and their
possible bene®cial and negative effects on the quality
of dairy products. International Dairy Journal 6:
755±768.
Rohm H, Eliskases-Lechner F and Braeuer M (1992)
Diversity of yeasts in selected dairy products. Journal of
Applied Bacteriology 72: 370±376.
Rosi J (1978) The ke®r (grain beverage) microorganisms:
the yeasts (Saccharomyces delbrueckii and Saccharo-
myces cerevisiae). Scienza e Tecnica Lattiero-Casearia
29: 59±67.
Seiler H and KuÈmmerle M (1997) Die Hefen¯ora von
Handelske®r. Deutsche Milchwirtschaft 48: 438±440.
Wyder MT (1998) Identi®cation and Characterization of
the Yeast Flora in Kefyr and Smear-Ripened Cheese:
Contribution of Selected Yeasts to Cheese Ripening.
PhD thesis, ETH, ZuÈrich, Switzerland.
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