The Good, the Bad, and the Ugly:
Tales of Mold-Ripened Cheese
SISTER NOLLA MARCELLINO, O.S.B.,1 and DAVID R. BENSON2
1
Abbey of Regina Laudis, Bethlehem, CT 06751; 2Department of Molecular and Cell Biology,
ABSTRACT The history of cheese manufacture is a natural
history in which animals, microorganisms, and the environment
interact to yield human food. Part of the fascination with cheese,
both scientically and culturally, stems from its ability to assume
amazingly diverse avors as a result of seemingly small details in
preparation. In this review, we trace the roots of cheesemaking
and its development by a variety of human cultures over
centuries. Traditional cheesemakers observed empirically that
certain environments and processes produced the best cheeses,
unwittingly selecting for microorganisms with the best
biochemical properties for developing desirable aromas and
textures. The focus of this review is on the role of fungi in cheese
ripening, with a particular emphasis on the yeast-like fungus
Geotrichum candidum. Conditions that encourage the growth of
problematic fungi such as Mucor and Scopulariopsis as well as
Arachnida (cheese mites), and how such contaminants might be
avoided, are discussed. Bethlehem cheese, a pressed, uncooked,
semihard, Saint-Nectaire-type cheese manufactured in the
United Sates without commercial strains of bacteria or fungi, was
used as a model for the study of stable microbial succession
during ripening in a natural environment. The appearance of
fungi during a 60-day ripening period was documented using
light and scanning electron microscopy, and it was shown to be
remarkably reproducible and parallel to the course of ripening of
authentic Saint-Nectaire cheese in the Auvergne region of
France. Geotrichum candidum, Mucor, and Trichothecium
roseum predominate the microbiotas of both cheese types.
Geotrichum in particular was shown to have high diversity in
dierent traditional cheese ripening environments, suggesting
that traditional manufacturing techniques selected for particular
fungi. This and other studies suggest that strain diversity arises in
relation to the lore and history of the regions from which these
types of cheeses arose.
The history of cheese manufacture is a natural history
in which animals, microorganisms, and the environment
interact to yield human food. Part of the fascination with
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University of Connecticut, Storrs, CT 06269-3125
cheese both scientically and culturally stems from its
ability to assume amazingly diverse avors as a result of
seemingly small details in preparation. These details
have been discovered empirically and independently by a
variety of human populations and, in many cases, have
been propagated over hundreds of years.
Cheeses have been made probably as long as mam-
mals have stood still long enough to be milked. In
principle, cheese can be made from any type of mam-
malian milk. In practice, of course, traditional herding
animals are far more effectively milked than, say, moose,
although moose cheese can be found in Sweden today.
Unfortunately, the price of moose cheese, not to mention
the scarcity of cooperative moose, dooms such enter-
prises to a niche market. The more common mammals
whose milk is used to produce cheese include cows,
sheep, goats, and, to a lesser extent, bison, camels,
horses, yaks, and llamas, depending on where one lives
in the world.
Words associated with cheesemaking have equally
ancient roots. The ancient Greeks called the wicker
cheese basket used for draining whey from the curds a
formos, which became forma to the Romans, from
which the modern Italian formaggio and French
fromage were derived (1). The English cheese is
Received: 13 April 2011, Accepted: 2 September 2011,
Published: 31 October 2013
Editor: Catherine W. Donnelly, University of Vermont, Burlington, VT
Citation: Marcellino N, and Benson DR. 2013. The good, the bad, and
the ugly: tales of mold-ripened cheese. Microbiol Spectrum 1(1):
CM-0005-2012. doi:10.1128/microbiolspec.CM-0005-2012.
Correspondence: Sister Nolla Marcellino, O.S.B., mnoella@
sbcglobal.net
2013 American Society for Microbiology. All rights reserved.
1
Marcellino and Benson
derived from the West Germanic reconstruct *kasjus via
the Old English cyse and Latin caseus. Their roots are in
the proto-Indo-European reconstruct *kwat-, to ferment
or become sour.
Human survival has a link to fermentation, the pro-
cess by which yeasts and bacteria degrade organic
compounds to by-products such as lactic acid and al-
cohol and transform perishable foods into preserved
products. In the case of cheese, the nal fermented
product is safe, durable, and, due to a reduced volume,
transportable, a feature upon which nomadic cultures
depend (2). Historically, cheese was often reserved for
use as a source of protein and fat during periods of low
milk production due to lactation cycles of animals (3) or
changes in climatic conditions (4).
Cheesemaking emerged as a by-product of dairying.
Domesticated animals played a role in Neolithic (4000
BCE) farming communities worldwide, but there has
been much conjecture as to whether they were used only
as sources of meat or were also exploited for secondary
products such as milk (57). Clear evidence of
cheesemaking is found in southern Mesopotamia, an
area long recognized as the cradle of civilization,
where great centers of religion and culture emerged in
the late fourth millennium BCE (8). Mesopotamia is the
Greek word for between the rivers, and it was here
along the banks of the Tigris and the Euphrates in the
Fertile Crescent that many believe that dairying
progressed to actual cheesemaking (9), as evidenced by
depictions of milking and processing of milk products
(10). Eighteen to twenty varieties of cheese have been
documented in the ancient Near East, including a-
vored, sweetened, sharp, crushed, and white (11).
Evidence for dairying in other ancient cultures has
been found in temperate Europe and in Mediterranean
regions, in the form of pottery. Shards of ceramic sieves
have been found at Linear Pottery sites distributed
across temperate Europe from the Ukraine to France and
from Hungary to the Baltic Sea (12, 13). The sieves were
probably used to drain whey from curds and suggest
that dairying took place in temperate Europe as early
as ca. 5400 BCE, the radiocarbon date for the Linear
Pottery culture in Hungary (14). More direct evidence
for prehistoric dairying has been obtained through the
eld of biomolecular archaeology, which has allowed
scientists to move beyond speculation based only on
reconstructed pottery vessels. Copley et al. (5) analyzed
fat residues from pottery from prehistoric sites using
fatty acid composition, double-bond positions, tri-
acylglycerol distribution, and 13 enrichment values
to evaluate the origins of the fats in relationship to
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domesticated animals. Of the 237 pot shards analyzed,
22% yielded dairy fat, indicating that dairying was in-
deed an important aspect of farming in the British Iron
Age. In other work, Craig et al. (15, 16) characterized
lipids and proteins adsorbed to late Bronze Age and Iron
Age ceramics from the island of South Uist in the Outer
Hebrides. The authors used gas chromatography-mass
spectrometry and gas chromatography combustion iso-
tope ratio mass spectrometry to identify the source of
lipids on the pottery. Twenty-ve of 39 shards tested
showed adsorbed lipids with a high abundance of satu-
rated fatty acids, especially 18:0, indicating that the
lipids were of animal origin. The ratio of 13C18:0 to
13C16:0 values further indicated that the animals were
ruminants. They also developed an immunological de-
tection method, the digestion-and-capture immunoas-
say, to extract and characterize the 2,500-year-old
proteins adsorbed to the ceramics. Their nding of bo-
vine -casein adsorbed to pottery shards was correlated
with the large number of neonatal cattle remains at the
site, evidence that calves were culled to sustain a dairy
economy. Maintaining lactating cattle with only a few
calves present in a herd implies that the cows were
milked and a large milk supply was transformed into
dairy products that could be stored and preserved. The
authors note the example of bog butter that has been
preserved and found in other sites where there is also
evidence of calf culling. Bog butter, a whitish, grey waxy
substance dating back to 400 BCE to 500 CE, has been
found in bogs in Ireland and Scotland. During the Iron
Age, butter was preserved in two-piece wooden kegs
buried in the anaerobic and cool conditions of the bogs
(17). Cronin et al. (18) analyzed the fatty acid content of
prehistoric bog butter in comparison to present-day Irish
fresh butter using gas-liquid chromatography and gas
chromatography-mass spectrometry. They found that
fatty acids are present in modern butter in the form of
triglycerides, while free fatty acids, in particular free
long-chain saturated fatty acids, predominated in the
bog butter due to hydrolysis. Proteolysis in the butters
was also studied using size exclusion high-performance
liquid chromatography and ion-exchange chromatog-
raphy. Not surprisingly, the bog butter consisted pri-
marily of small peptides (<0.5 kDa) and free amino acids
with little or no intact proteins, in comparison to mod-
ern butter, in which caseins and whey proteins, ranging
in size from 14 to 30 kDa, remain. The authors con-
cluded that proteolysis in the bog butter was probably
due to microbial activity in the bog environment.
In ancient Greece and Rome, cheese was an essential
and popular food. Fresh milk was never imbibed; in fact,
 The Good, the Bad, and the Ugly: Tales of Mold-Ripened Cheese
the barbarians were called, somewhat derisively, milk-
drinkers (19). The Scythians, the horse-riding tribes of
the Eurasian steppes, were called mare-milkers by
Homer (ca. 1184 BCE) and consumed fermented mares
milk and cheese called hippake (20). Because of the
abundance of olive oil in the Mediterranean, butter was
not used. The Greek geographer Strabo (66 BCE25 CE)
viewed the mountain people of the Iberian Peninsula
with distaste because they lived on goat meat and acorn
bread, drank beer rather than wine, and used butter
instead of olive oil (21). Varro (11628 BCE), in his
treatise on Roman farm management, wrote, Cheese
made of cows milk is the most agreeable to the taste, but
the most difcult to digest: next, that of ewes milk,
while the least agreeable in taste, but the most easily
digested, is that of goats milk (22). Columella (4 BCE
ca. 70 CE) wrote of all aspects of cheesemaking in his
work on agriculture, Rei Rusticae (23). The principles he
enunciated in the 1st century could be applied today. He
described whey as the acid liquid which should be
pressed out of the cheese as quickly as possible (23).
Today, we know that excess whey that contains lactose
in pressed cheeses can cause postacidication" of the
curd, resulting in bitterness (24). Homer (ca. 1184 BCE)
was familiar with aspects of cheesemaking, as evidenced
by the detail with which he describes the scene in the
cave of the cyclops Polyphemos in his epic poem the
Odyssey. Upon entering the cave, Odysseus notices
vessels swimming with whey and willow baskets
that were heavy with cheese (25) which he and his men
eat while awaiting their host. However, the cyclops, a
giant dairy farmer (26), is also a savage monster
who despises the laws of hospitality to the point that he
devours guests in his own home, if indeed one can call
his cave a home (27). While his prisoners cower, the
cyclops performs his dairy chores twice a day: he milks
his sheep, adds g juice to form a curd (g juice contains
the protease cin, which acts to coagulate the milk), and
drains the curd in willow baskets. Odysseus and his
surviving men eventually escape by blinding the cyclops
and riding out of the cave underneath the bellies of
his rams.
Given the coincidence between dairying, cheesemaking,
and civilization, it is not surprising that Kamber and
Terzi regard cheese as one of the primary symbols of
mankinds passage into civilization (28), and tech-
niques for its production, unique to each culture, were
handed down from generation to generation as a cultural
inheritance (29). In modern times, the establishment of
the European Economic Community has prompted a
new self-awareness of each countrys regional products
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in Europe that reect the cultural and environmental
characteristics of particular locales (30). In the developed
world, there has been a trend towards decentralizing
food production and a renewed interest in local foods
that are identied with particular rural areas and eth-
nicity, to revitalize agriculture in economically depressed
or depopulated regions (31).
Legislation to guarantee the territorial authenticity
and vintage of a product rst arose in France, where, in
1411, King Charles VI granted the people of Roquefort-
sur-Soulzon a monopoly on Roquefort-style cheese and
in 1666 the court of Toulouse applied a judicial text to
Roquefort cheese, making it illegal for other villages to
assert that they produced the cheese. These declarations
simply recognized a cheese type that had been produced
since at least Roman times. Subsequent legislation
created the appellation dorigine contrle (AOC) to
protect traditional products that arose within the
boundaries of a rural community and its specic envi-
ronmental context (32). Many European artisanal
cheeses are now marked with the seal of the protected
designation of origin (PDO), a denomination established
by the European Union to recognize and protect agri-
cultural products whose quality or characteristics are
essentially or exclusively due to a particular geographi-
cal environment with its inherent natural and human
factors and the production, processing and preparation
of which take place in the dened geographical area
(33). As early as 500 CE, Cassiodorus (480575 CE),
Praetorian Praefect of the Ostrogothic king Theodoric,
evoked terroir when he described the banquets in Rome
which included cheeses made from the milk of herds
grazing in the lush pastures and forests on Mount Sila in
southern Italy:
and we praised the wines of Bruttii and the cheese of the
district around Mount Sila. The cheese, which retains in its
pores the milk which has been collected there, recalls by its
taste the fragrant herbs upon which the cattle have fed; by
its texture it reminds us of the softness of oil, from which it
differs in colour by its snowy whiteness. Having been carefully
pressed into a wide cask and hardened therein, it retains per-
manently the beautiful round shape which has thus been given
to it.
Variae Epistolae, Book XII (34)
The concept of a geography of taste (31) has now
taken hold on both sides of the Atlantic. Reacting to the
globalized food market and suspicious of industrial food
processing techniques, consumers increasingly demand
to know the source of their food and are seeking a more
direct link to the producers (35). Hence, sustainable
3
Marcellino and Benson
agriculture, which includes the concept of reversing the
trend of disappearing farmland, is on the rise.
CHEESE RIPENING
Cheese is milk that has grown upit is preeminently the food
of manthe older it grows the more manly it becomes, and in
the last stages of senility it almost requires a room to itself.
The Epicures Companion, Edward Bunyan (18721939) (36)
Throughout the centuries, a blend of empirical obser-
vation and isolated cultural development has helped
sculpt the diversity of cheeses we know today. Flavor
diversity is created by the biochemical activities of
microorganisms degrading the curd and providing
avors and aromas absent from the initially bland young
cheese (37, 38, 39). Pasteurizing the milk used for
cheesemaking allows for more uniformity of the nished
product and improves food safety but removes much of
the indigenous microbiota of milk that imparts distinc-
tive avors (40). The large-scale industrialization of
cheesemaking has caused a loss of traditional techniques
and ripening venues, including natural caves where di-
verse populations of fungi and bacteria have developed
sometimes over centuries. In response, some micro-
biologists have been isolating and characterizing the
members of microbial populations originating in tradi-
tional cheeses unique to specic geographical regions
(39, 40, 41, 42).
In one such project, 4,379 isolates of wild-type lactic
acid bacteria were characterized from 35 products, in-
cluding 24 artisanal cheeses made from cow, sheep,
goat, and buffalo milk using traditional techniques in
southern Europe. Not surprisingly, tremendous vari-
ability in acid and exopolysaccharide production, pro-
teolytic activity, and bacteriocins of the isolates was
discovered. A conclusion was that each cheese was an
ecosystem in itself (43). Cheesemakers for centuries
have exploited such extraordinary diversity in the cheese
cave or cellar where a cheese is aged.
Even with this diversity, cheese microorganisms tend
to follow a similar script during ripening. Three primary
biochemical processes take place: fermentation of milk
sugar (lactose) to lactic acid; hydrolysis of lipids to fatty
acids; and breakdown of casein to peptides, amino acids,
and ammonia. Amino acids and fatty acid derivatives
are precursors to avor compounds. Their metabolism
plays a key role in determining the aroma of a cheese
(44). These processes lead to the desirable, and some-
times undesirable, sensory qualities of avor, texture,
and appearance (45), and they are mainly due to the
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enzymes of the bacteria and fungi growing in and on the
cheese that use various components of milk for their
own nutritional requirements.
The normal microorganisms involved in cheese rip-
ening include lactic acid bacteria naturally present in the
milk and/or added in commercial starter cultures. Lactic
acid bacteria, the live cultures with which most people
who eat yogurt are familiar, quickly catabolize lactose to
lactic acid and lesser amounts of acetic acid in the critical
step of lowering the pH of the curd during cheese pro-
duction. Other microorganisms play a more or less im-
portant role in ripening depending on the type of cheese.
They include nonstarter indigenous lactic acid bacteria,
other bacteria such as the aerobic surface-dwelling
Brevibacterium linens, yeasts, and lamentous fungi
(46). The microbial populations that develop on the
surface of cheese can be quite complex depending on
where they are ripened and how they were prepared.
Some cheeses have traditionally been aged in jars and
earthenware vessels buried in sand and soil (28), sealed
pits called fosse dug in volcanic rock (47, 48), and/or in
caveseach environment bringing the cheese in prox-
imity to soil and diverse types of microorganisms (49,
50). Members of soil fungal genera such as Alternaria,
Chrysosporium, Geotrichum, Mucor, and Penicillium
are commonly found on cheese ripened in cave en-
vironments (51, 52).
Scott observed that Fermentation organisms are
ambient in the environment, whether humans make use
of them or not (53). And indeed humans have. If a
cheesemaker observed empirically that certain con-
ditions produced the good cheeses, he or she tried to
reproduce or maintain the environment that by default
selected for strains with the best biochemical properties.
Afneur Pierre Androut says it this way: Our pre-
decessors thought with reason that the natural agents in
the environment conditioned the personality of cheeses
and marked them with the indelible sign of vintage and
territory (54).
THE STRAIN YOU LOVE MAY BE YOUR OWN
A case study for how novice cheesemakers can come to
terms with the cheesemaking process, indigenous
microorganisms, and the terroir of a region may be told
through the experiences of one of us (N.M.), who
established a small artisanal cheesemaking facility at the
Benedictine Abbey of Regina Laudis in Litcheld
County, CT.
When one thinks of dairying in New England, Con-
necticut is not the rst state to come to mind.
 The Good, the Bad, and the Ugly: Tales of Mold-Ripened Cheese
Connecticuts farmland is disappearing faster than that
of any other state; approximately 9,000 acres of farm-
land are lost to development every year.
As an aside, we note that the loss of farmland is at odds
with Connecticut history. Contributing to the westward
expansion of agriculture in the United States, The
Connecticut Yankee brought a cheese hoop with him and
wherever he went made cheese (55). The Statistical
Account of the Town of Bethlehem for 1812 by the
Reverend Azil Backus reports that there are commonly
sent to market 7,000 lb. of butter and 30,000 lb. of
cheese (56). The rst cheese factory in America was
built by Lewis Mills Norton in 1844 in Goshen, another
town in Litcheld County known for its lush farmland
and pastures. The pineapple cheese produced there
earned its name from the pineapple-shaped wooden
mold into which Norton pressed a soft Cheddar-type
cheese (57). By the middle of the 19th century, Litcheld
County was making nearly 3 million pounds of cheese a
year. Pineapple cheese was rst transported to the port
at New Haven for distribution, but by 1884 the demand
for the cheese was so great that the plant was eventually
moved to Attica, NY, for better access to the railroads.
From this period, cheesemaking in Connecticut began its
decline.
Cheesemaking returned in earnest at Regina Laudis in
1977 when a third-generation cheesemaker from
Czallier in the Auvergne, France, came as a visitor. She
brought with her the traditional methods of making a
Saint-Nectaire-type cheese that she had learned from her
grandmother. After 2 years and many disasters, a con-
sistently good cheese was developed, which the abbey
named Bethlehem cheese in deference to the AOC des-
ignation of Saint-Nectaire. Bethlehem cheese is a pressed,
uncooked, semihard, mold-ripened cheese made from the
fresh milk of the abbeys Dutch Belted cows, a heritage
breed whose milk, high in protein, fat, and total solids, is
excellent for cheesemaking. The milk is not pasteurized,
and commercial cultures of starter or fungi are not added
during production or ripening.
Traditionally, the cheeses ripen on straw mats for 60
days in the cellar of a housethe cave. During this typical sporangiospores in spherical, black sporangia.
time, the appearance of fungi growing naturally on the
cheese rinds is remarkably reproducible and predictable
and parallels the course of ripening of authentic Saint-
Nectaire cheese in the Auvergne (58).
Fungal genera from Bethlehem cheese and raw milk
Saint-Nectaire cheeses from the Auvergne region of
France were found to be similar and included Geotrichum
candidum, Mucor, and Trichothecium roseum. The con-
clusion was that similar means of preparation lead to
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similar microbial successions during ripening. Bethlehem
cheese thus was used as a model for the study of stable
microbial succession during ripening in a natural envi-
ronment (58).
Microbial succession on the ripening cheese over 60
days was monitored by scanning electron microscopy
and light microscopy; the latter used stained parafn
sections to view cross-sections of the rind at various
stages (58). When the cheese is removed from the cheese
press, it has a rubbery consistency, very little avor, and
no discernible rind; indeed, no surface microorganisms
were visible by scanning electron microscopy. In the
curd, however, pockets of gram-positive cocci and yeasts
had developed, indicating that bacterial growth and
fermentation begin within the curd soon after formation.
The morphology of all cells within an individual pocket
was the same, suggesting derivation from single cells.
At 48 h, the cheese surface is covered with a lawn of
budding yeast (Fig. 1A). The number of yeast CFU on
the nascent rind increases at least 40-fold by day 4 and
stabilizes from day 10 to the end of ripening. The
number of lactic acid bacteria on the rind and in the curd
remains fairly constant throughout ripening. The sharp
increase of yeasts by day 4 is probably due to their
ability to use some of the lactic acid produced by the
lactic acid bacteria, leading to an increase in the pH of
the rind. In many cheeses, this deacidication by yeast
prepares the surface for the growth of acid-sensitive
aerobic bacteria such as Brevibacterium linens (59).
Strains of Streptococcus cremoris are the most abundant
lactic acid bacteria within Bethlehem cheese, probably
derived from the raw milk used in production. In con-
trast to the case with the rind, yeast counts remain low
and fairly stable within the curd throughout the ripening
process. Strains of Debaryomyces and Torulopsis were
identied, with the latter predominating in the interior of
the cheese. Torulopsis ferments lactose to the neutral
ethanol that is important in low-acidity cheeses such as
Saint-Nectaire (60, 61).
By day 6 of ripening, the surface of Bethlehem cheese is
covered by the whitish-gray Mucor, identied by its
Fig. 1B shows Mucor sporangia and hyphae on the sur-
face of the cheese. By day 9, the sporangiospores release
from the sporangia and the cheese takes on a grayish-
brown velvety coat composed of mycelia and spores from
Mucor entwined with Geotrichum candidum and drying
of the cheese curd surface. G. candidum begins to dom-
inate around day 4 of ripening and covers the cheese with
a white powdery coat. G. candidum plays an important
role in the ripening of soft cheeses such as Camembert
5
Marcellino and Benson
FIGURE 1 Scanning electron micrographs of Bethlehem
cheese during ripening. (A) Micrograph of cheese at day 2 of
ripening showing multilateral budding yeasts embedded
within the cheese surface. Buds and bud scars are clearly
visible. Bar, 1 m. (B) Micrograph of the cheese surface at day 6
showing sporangia and hyphae of the zygomycete Mucor. Bar,
10 m. (C) At day 59, the V shapes of coryneform bacteria can
be distinguished at a high magnication beneath a mass of
fungal debris on the cheese rind. Bar, 1 m. Panels A and B are
reproduced with permission from reference 58. doi:10.1128/
microbiolspec.CM-0005-2012.f1.
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and semihard cheeses such as Saint-Nectaire and Re-
blochon. Its lipases and proteases release fatty acids and
amino acids and peptides involved in avor develop-
ment, and its aminopeptidases reduce bitterness im-
parted by low-molecular-weight peptides in cheese (62).
While the growth of Mucor is typical on traditional
Saint-Nectaire and Tomme de Savoie cheeses (38, 63), it
can be a disaster on pte molle-type cheeses such as Brie
and Camembert.
By day 60, patches of pink mold can be seen high-
lighting the grayish coat left from the growth of Mucor.
This pink fungus has been termed the ower of the
molds and is valued by makers of Saint-Nectaire cheese.
Microscopically, the two-celled conidiospores that de-
velop from the tip of the vertical conidiophore are di-
agnostic of Trichothecium roseum (Pers.) Link 1809.
Identication T. roseum is conrmed by isolation of the
fungus and observation of conidiospore development as
occurring in short chains in basipetal succession from the
conidiophore using light microscopy (64). The contri-
bution of T. roseum to the taste of cheeses like Bethlehem
or Saint-Nectaire has not been studied, although the
fungus is known for its lipolytic activity (65).
By day 59 of ripening, a more complex microbial
community has settled in and stabilized on the surface of
the cheeses, including lamentous fungi and coryneform
bacteria that are rst evident by day 20 of ripening.
Species of Brevibacterium and Arthrobacter spp. can be
isolated from Bethlehem cheese based on cell morphol-
ogy and colony color. Fig. 1C shows the distinctive V
shape of coryneform bacteria that have developed be-
neath the fungal debris on the cheese surface.
Brevibacterium linens, the bacterium that predominates
in the mixed culture on the surface of smear-ripened
cheeses such as poisses, Mont dOr, Limburger, and
Taleggio, gives cheeses a characteristic reddish-orange
color (66). B. linens produces volatile sulfur compounds
from the degradation of L-methionine, which contributes
to the unique, sometimes pungent aroma of this class of
cheeses and, by the way, to foot odor (67).
The ripening of Bethlehem cheese is similar in pro-
gression to that of other mold-ripened cheeses, with
some of the same organisms involved, including the
lactic acid bacteria, Geotrichum candidum, and various
yeasts. The difference in taste and consistency is due to
the differences in microbial communities, which, in turn,
are selected for by differences in handling of the curd as
it is being prepared. The selection often occurs at the
level of salting of the cheese. Inhibition of G. candidum
can occur at 1.0 to 2.0% salt and of Mucor at 2.0 to
3.0%. Penicillium species are the most salt tolerant of
 The Good, the Bad, and the Ugly: Tales of Mold-Ripened Cheese
the three genera, growing well at 5.0% salt. Saint-
Nectaire-style cheeses are dry salted, and after 24 h in
the cheese press, the residual salt is rinsed off, allowing
for the growth of Geotrichum candidum, followed by
Mucor. In the production of the Spanish blue-veined
cheese Cabrales, during which no starter or fungal
cultures are added, the drained cheeses are covered with
a layer of coarse salt which is not washed off and is kept
at room temperature for 10 to 15 days. The cheeses are
subsequently placed in natural caves, where the salt
content and high humidity in the environment encour-
age the growth of Penicillium roqueforti (68). Hence, the
cheesemaker directs the microbial population to differ-
ent outcomes.
FRENCH LESSONS IN BIODIVERSITY
The fact that the same types of fungi are found on tra-
ditional Saint-Nectaire from France and Bethlehem
cheese from the United States indicates that the critical
microorganisms are present naturally in the environ-
ment and that the traditional methods of preparation
and ripening determine the nished product. At the
same time, the genetic and biochemical diversity of
Geotrichum populations in the milk and curd and on
cheeses ripened in traditional caves of France in six
major cheesemaking regions was found to be immense
(38). Saint-Nectaire, Reblochon, Mont dOr, and Tami
were looked at with regard to strain diversity in relation
to the lore and history of the regions from which they
arose. The conclusion from that study indicated that a
tremendous amount of biochemical and genetic diversity
existed within the Geotrichum candidum clade. Further,
such diversity can partly be explained by the empirical
selection of cheesemaking techniques that favored the
development of the best strains in local contexts. A
corollary is that manufacturing practices aimed at pro-
ducing a uniform cheese type would necessarily lead to
the loss of specic local strains.
Geotrichum candidum: The Good
The loss of diversity has implications for cheesemakers.
Among the good, the bad, and the ugly fungi,
Geotrichum candidum stands as one of the best and thus
receives particular attention in this review, but it should
be noted that other microbial groups would paint a
similar picture.
G. candidum contributes to the ripening of nearly all
surface mold-ripened cheeses: pte molle such as Brie
and Camembert, semihard cheeses which include Saint-
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Nectaire, Tomme de Savoie, and Reblochon, and the class
of smear cheeses to which Livarot, Limburger, and
Taleggio belong (69, 70). Pottier et al. (71) evaluated the
importance of G. candidum to the French dairy industry,
calculating that G. candidum was present in about
600,000 tons of French cheese, one-third of total pro-
duction. They concluded that in 1 year, French consumers
ate close to 8 kg (18 lb) per person of cheese containing
G. candidum. Fifteen years ago, G. candidum may not
have been commonly discussed in the United States, yet
now at the annual American Cheese Society conference, it
is not unusual to hear artisanal cheesemakers comparing
their strains of G. candidum, or geo. Both wild-type
strains and commercial cultures affect the appearance and
aroma of cheeses. In catalogues, starter strains are
presented according to characteristics that might interest
the cheesemaker: colony color, morphology (yeasty or
moldy), biochemical properties (having known pro-
teolytic or lipolytic activity), and capacity for neutralizing
the curd, which affects the subsequent microbial ecology
of the cheese.
Growth of G. candidum is generally observed on
the cheese surface around the third day of ripening. In
some cheeses, such as Saint-Marcellin, the main role of
G. candidum is in the presentation of the nished
product: the organism covers the cheese surface with a
uniform white velvety layer, sometimes called a jolie robe
(pretty dress) (72). On other cheeses, G. candidum is not
obvious at the end of the ripening period. On ripened
Camembert or Brie, G. candidum is covered by a white
layer of Penicillium camemberti which rst appears
around day 6 of ripening (73, 74). G. candidum is the
only lamentous fungus on Reblochon and, mixed with
Brevibacterium linens, creates an orange-white crust (67,
75). In contrast, G. candidum is part of the more com-
plex microbiotas on the surfaces of Saint-Nectaire and
Tomme de Savoie (62). Figure 2A shows a layer of
G. candidum on the rind of a 10-day-old Tomme de
Savoie that will be covered by a gray layer of Mucor later
in ripening.
Morphologically, G. candidum is described as a
yeast-like fungus because it appears microscopically
with both single-cell stages and laments (72). Figure 2B
shows a light micrograph of a G. candidum strain iso-
lated from a 4-day-old Bethlehem cheese. Two cell types
can be seen: arthrospores and short septate hyphae.
During the process of asexual reproduction, the hyphae
fragment and this process, called disarticulation,
produces thallic-arthric conidia, called arthrospores (76)
the cylindrical cells seen interdispersed among the
hyphae in Fig. 2B. The scanning electron micrograph in
7
Marcellino and Benson
FIGURE 2 The yeast-like fungus Geotrichum candidum. (A)
G. candidum on the rind of a 10-day-old Tomme de Savoie.
(B) Light micrograph of arthrospores and septate hyphae of a
wild-type G. candidum strain isolated from a 4-day-old
Bethlehem cheese. Bar, 10 m. (C) Scanning electron micro-
graph of the surface of a Reblochon ripening shelf showing
cylindrical G. candidum arthrospores and bacteria. Bar, 5 m.
Reproduced with permission from reference 77 (micrograph
by Romain Briandet, UMR-UBHM, Institut National de la Re-
cherche Agronomique et Ecole Nationale Suprieure des In-
dustries Agricoles et Alimentaires, Massy, France). doi:10.1128/
microbiolspec.CM-0005-2012.f2.
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Fig. 2C (77) shows G. candidum arthrospores, roughly
10 m in length, on the surface of a wooden Reblochon
ripening shelf.
Yeast-like strains are generally characterized by
cream-colored colonies, very similar to yeast colonies on
agar medium. On a microscopic level, these strains pro-
duce many arthrospores and very few vegetative hyphae
(62). They generally have an optimal growth tempera-
ture of 22 to 25C, show acidifying activity, and are
weakly proteolytic (78, 79). More lamentous strains are
characterized by white colonies, more or less furry or
felt-like, which have vegetative hyphae predominating
with very little sporulation (62). These fungus-like strains
grow best at 25 to 30C, have alkalizing activity, and are
highly proteolytic.
Classication of G. candidum may not necessarily
interest cheesemakers, yet the commercial strain biotype
may strongly inuence the texture, cohesiveness, and
thickness of the rind. The taxonomy of Geotrichum
has a complex history and is still evolving. About 100
synonyms have been used for the same taxon since 1809,
when Link designated the genus as Geotrichum, with
G. candidum as the only species (79). Previous names,
such as Oidium lactis, assigned in 1850 when it was
isolated from milk by Fresenius, reveal a historical con-
nection to milk and dairy products (80). Traditional
French cheesemakers to this day sometimes refer to
G. candidum as Oidium. The microorganism was re-
classied as Oospora lactis in 1880 and nally placed in
the genus Geotrichum. G. candidum was included in
Barnetts rst and second editions of Yeasts: Char-
acteristics and Identication and classied as a fungal-
like yeast (81, 82).
Until the 1990s, fungal and yeast classication was
based on observable (phenotypic) characteristics such as
morphology, physiology, and biochemical properties
(83). Together with its distinctive morphology, growth
on D-xylose and lack of growth on cellobiose and
maltose as sole sources of carbon are used in identifying
Geotrichum to the species level (72). Despite its ability to
live on cheese, G. candidum cannot use lactose as a
carbon source, whereas assimilation of lactic acid is
variable (62, 84). Phenotypic variability presents a dif-
cult task for identifying each new strain (85).
Over the past 10 years, advances in molecular
methods for identifying and typing strains have led to
revisions in classication. G. candidum had been clas-
sied as the imperfect form, or anamorph, of the yeast
Galactomyces geotrichum in 1986 by de Hoog et al.
(86). However, taxonomic revisions of lamentous
fungi that reproduce by arthric conidiogenesis, based on
 The Good, the Bad, and the Ugly: Tales of Mold-Ripened Cheese
internal transcribed spacer ribosomal DNA sequences
and nuclear DNA (nDNA)/DNA reassociation data, led
to renaming of the teleomorph (perfect form) Galac-
tomyces candidus (87, 88). Isolates of the anamorph
genus Geotrichum were identied to the species level by
using randomly amplied polymorphic DNA-PCR
(RAPD-PCR) (89). In our investigation into the biodi-
versity of G. candidum in French cheese, we differenti-
ated strains within species using RAPD-PCR (38). Other
approaches have used chromosomal pulsed-eld gel
electrophoresis to correlate genetic data with pheno-
typic characteristics such as morphology (90). The latter
approach showed that the fungus-like strains and those
with intermediate morphology had larger genomes than
yeast-like strains. A standardized protocol has been
proposed for identifying G. candidum that allows
identication and comparison between laboratories
(83). It uses PCR to distinguish between species of
Geotrichum and then differentiates strains within the
species using RAPD-PCR in conjunction with random
amplied microsatellite PCR. The GATA4 primer used
in random amplied microsatellite PCR allowed strains
to be grouped according to the ecological niche from
which they were isolated, such as food, plants, the en-
vironment, and human sources. The protocol can be
used to trace strains to their provenance and facilitates
tracking of strains throughout cheese production and
ripening.
G. candidum strains not only are part of the
microbiota of raw milk but also are found in en-
vironments as diverse as soil, barley, grass, and silage.
The optimum pH for growth is around 5.5, but they can
grow across pH values of 3.5 to 9.0 (91). G. candidum
strains are generally acid-tolerant microorganisms that
can create costly problems for citrus fruit and tomato
industries (92, 93). The organism is part of the natural
microbiota of barley and is used as an inoculum in malt
production to improve quality as well as to inhibit the
growth of Fusarium species. The latter cause the defect
of gushing in beer and can produce mycotoxins (94).
G. candidum possesses an array of enzymes with great
potential for biotechnological applications. Its lactate
oxidase has been used to determine lactate levels impli-
cated in heart disease (95). In toxic waste management,
G. candidum is used to decolorize azo dyes in industrial
waste water (96), and its peroxidases have shown high
efciency in decolorizing and biodegrading olive mill
wastewater, a pollutant generated by olive oil extraction
(97). In 2001, G. candidum gained notoriety as the
fungus that eats CDs when its enzymes bored holes
in the aluminum and polycarbonate layers of com-
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pact discs and corrupted the information stored in them
(98).
G. candidum is not an agent of food poisoning and is
generally regarded as safe. No food-borne disease has
been linked to the consumption of products containing
G. candidum, and food technologists handling the
microorganism are not are risk (71). However, some
G. candidum strains can act as opportunistic pathogens,
causing supercial or internal mycosis known as geo-
trichosis (90, 99).
Lipolytic enzymes contributed by milk, starter bac-
teria, and fungi help develop texture and release free
fatty acids, precursors to avor compounds in many
cheeses (100). From a biochemical standpoint, G.
candidums lipases are unique and contribute to avor
and texture development in several classes of cheese
(101, 102). One isoform of G. candidums lipases
exhibits a preference for cis (-9) unsaturated fatty acids
in triglyceride estersfatty acids such as oleic or linoleic
acid with a double bond at the ninth carbon position.
Fresno et al. (103) attribute the high concentration
of oleic acid in Armada goat cheese to the fact that
G. candidum predominates on the cheese during the rst
2 months of ripening. In addition, the lipase has been
used for biotransformations in the oil industry, such as
the synthesis of (S)-atenol and (R)-propanolol through
lipase-catalyzed hydrolysis of the intermediate O-acetyl
esters, and in pharmaceutical production for the
stereoinversions of arylethanols (104, 105).
Of the three main biochemical activities involved in
cheese ripeningproteolysis, glycolysis, and lipolysis
proteolysis is considered the most complex and impor-
tant (106). While proteolysis is a key to avor develop-
ment, it is also responsible for bitterness, a common
defect in cheese (107). Mold-ripened cheeses are espe-
cially prone to bitterness (108), as are cheeses with low
salt concentrations. High salt concentrations in cheese
may promote the aggregation of large peptides, making
them less accessible to degradation by proteinases into
bitter peptides (109).
Aminopeptidases of G. candidum strains have been
correlated with reduced bitterness in Camembert through
the breakdown of low-molecular-weight peptides released
by the proteinases of Penicillium camemberti (74). The
synergistic effects of the two fungi on protein catabolism
in Camembert are summarized in Fig. 3. Proteolysis in
cheese begins when the proteinases (chymosin) in coagu-
lant, milk (plasmin), and starter cultures hydrolyze casein
to high-molecular-weight peptides (Fig. 3A). The high-
molecular-weight peptides are then catabolized to low-
molecular-weight peptides by proteinases of starter
9
Marcellino and Benson

Casein The size of peptides (molecular mass < 6,000 Da), the
nature of the N-terminal amino acids, and the higher
Chymosin mean hydrophobicity of amino acids in a peptide chain
(coagulant) PROTEINASES
are factors implicated in bitter taste (109, 116). Peptides
A containing proline have been associated with bitterness in
(Lactic Acid Bacteria
Plasmin cheese, as have peptides with basic amino acids such as
starter cultures)
(milk)
lysine and arginine in the N-terminal position (117, 118,
119). In one study, a synthetic peptide with arginine in the
N-terminal position was 250 times more bitter than
High Molecular Weight Peptides caffeine (116). A high content of the hydrophobic amino
acid leucine in a peptide also increases bitterness (120).
PROTEINASES increasing Aminopeptidases, in cleaving the N-terminal amino acids
B (Penicillium camemberti) of a hydrophobic peptide, can reduce bitterness (121,
bitterness
122). To that end, aminopeptidases from Aeromonas
caviae and the fungi Rhizopus oryzae and Aspergillus
Low Molecular Weight Peptides sojae have been used to debitter protein hydrolysates (120,
123). The released free amino acids are precursors to a-
vor compounds, and their catabolism is signicant in
AMINOPEPTIDASES
(Geotrichum candidum) mold-ripened cheeses (114). Through reactions such as
reduction of deamination, transamination, and decarboxylation, the
C CARBOXYPEPTIDASES amino acids are converted to alcohols, carbonyls, or short-
bitterness
(Penicillium camemberti, chain hydrocarbons (Fig. 3D) (106). Deamination of
Geotrichum candidum)
glutamate, aspartate, leucine, phenylalanine, and methi-
onine by G. candidum has been reported (124). The
breakdown of sulfur-containing amino acids such as me-
Amino Acids thionine by G. candidum produces desirable garlic or
cabbage aroma notes characteristic of some pte molle
Deamination
and smear cheeses (125, 126). Molimard (127) correlated
D the cabbage aroma of traditional Camembert with the
Decarboxylation concentration of isovaleric acid, a derivative of leucine via
Transamination
Strecker degradation, which has also been found in Swiss
cheese (128). Jollivet et al. grouped eight G. candidum
strains according to their production of methyl ketones,
secondary alcohols, phenol, phenylethanol, and dimethyl
Volatile Aroma Compounds disulde, when grown in milk and cheese media (129). All
FIGURE 3 Schema for proteolysis in Camembert cheese.
strains produced the compound 2-methylpropan-3-ol, an
doi:10.1128/microbiolspec.CM-0005-2012.f3. alcohol previously not identied in cheese.
We studied the extracellular aminopeptidases of
13 wild-type G. candidum strains isolated from
bacteria and secondary microorganisms (110, 111). Camembert, Mont dOr, Reblochon, Saint-Nectaire,
G. candidum presumably does not have to hydrolyze Coulommiers, and Bethlehem cheeses chosen as rep-
caseins into peptides, as these can be provided by the resentatives of major groups in our RAPD-PCR study.
proteolytic activity of other organisms. Boutrou et al. We found that differences in the enzyme activity (the hy-
(112) have pointed out that although G. candidum is ca- drolysis of p-nitroanilide derivatives of lysine, arginine,
pable of using casein and peptone, in cheese its role is leucine, methionine, and alanine) correlated with the
mainly peptidolytic rather than proteolytic. Bitterness complexity of microbial populations characteristic of
in Camembert emerges at this step when the proteinases of these types of cheeses (unpublished data). Three strains
P. camemberti degrade high-molecular-weight peptides to isolated from Saint-Nectaire and 1 from Bethlehem cheese
low-molecular-weight peptides (Fig. 3B) (113). The s1- clearly had the highest activity of the 13 isolates tested.
and -caseins have hydrophobic regions that, when re- The strain with the highest activity of the 13 tested in the
leased by proteinases, increase bitterness (106, 114, 115). release of leucine, alanine, and methionine was the only

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 The Good, the Bad, and the Ugly: Tales of Mold-Ripened Cheese
American strain, isolated from a 45-day-old Bethlehem
cheese. The activity of strains isolated from cheese with
complex fungal populations was in sharp contrast to that
of the 4 strains isolated from Reblochon from two dif-
ferent cheesemaking facilities. Reblochon has a less com-
plex microbiota than Saint-Nectaire. The gray covering
seen on Saint-Nectaire and Bethlehem cheeses is due to the
zygomycete Mucor, strains of which are known to be
highly proteolytic. Indeed, the acid proteinases produced
by Mucor pusillus and Mucor miehei which cleave the
Phe105Met106 peptide bond of -casein have been used
commercially for microbial and genetically engineered
rennet for years due to the shortage of calf rennet (130).
Saint-Nectaire strains may thus be selected for by the ac-
cumulation of peptides released by Mucor proteinases
that are, in turn, metabolized by the aminopeptidase of
G. candidum. In contrast, a strain that was isolated from
Saint-Nectaire curd before Mucor had developed showed
little activity.
Besides having a less complex fungal population, Re-
blochon is dominated by the bacterium Brevibacterium
linens, which is highly proteolytic, possessing both
proteinases and aminopeptidases, and has been used
as an adjunct in cheesemaking to accelerate ripening
(106, 131). The pathways of methionine catabolism
by B. linens have been investigated because the by-
product, methanethiol, gives Reblochon and other
surface-ripened cheeses their characteristic avor (132,
133). Since B. linens has aminopeptidases, strains of
G. candidum with high aminopeptidase activity may not
be selected for in Reblochon.
Not all strains of G. candidum are equal. Some cause
defects in cheese described as greasiness, bitterness, and
a strong yeasty avor (134). On some cheese rinds, the
proteolytic activity of G. candidum causes a wrinkly
appearance that is referred to as peau de crapaud, or
toad skin, by French cheesemakers. This texture is
desirable on some cheeses, such as goat cheeses; but in
excess causes a slippery rind. When the cheese is turned
during ripening, the skin can separate from the cheese
surface, thus ruining the integrity of the rind. To stop
excess growth, the cheesemaker may lower the temper-
ature and/or humidity of the production area and cave.
Due to G. candidums sensitivity to salt (inhibition can
occur at 1.0 to 2.0% salt), using a higher concentration
of salt or changing the salting method from brining to
dry salting may resolve the problem of excessive growth
(108). Attention must also be paid to the amount of
inoculum. Le Bars-Bailly et al. (108) note that one spore
of G. candidum per kilogram of curd introduced during
cheese production can lead to a population of 105
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arthrospores per gram of cheese after 21 days of ripening
at 10C. The threshold for a yeasty taste is between 4
103 and 104 G. candidum spores per gram of curd.
The neutralization of the cheese surface by G.
candidum and by other yeasts prepares the cheese sur-
face for acid-sensitive bacteria, such as Brevibacterium
linens, which follow in the later stages of ripening (67).
B. linens grows best at a neutral pH of 7.0 (135). In
experimental Camembert-type cheeses inoculated with
G. candidum, the pH of the rind reached 7.0 at the end
of ripening, whereas the pH of control cheeses remained
at about 4.8 (112).
The increase in pH is generally attributed to the
catabolism of lactic acid produced by starter bacteria
to carbon dioxide (CO2) and H2O (72, 136). In our
diversity studies, the four wild-type G. candidum strains
isolated from Reblochon were all positive for assimila-
tion of lactate as the sole source of carbon. The four
producers used yogurt as the source of starter culture,
perhaps selecting for strains capable of using lactate as a
source of carbon.
Release of ammonia also plays a role in neutralizing
the cheese environment. Studies of the growth kinetics of
G. candidum on peptone-based medium with and
without lactate showed that G. candidum preferred
peptones as a carbon source, even when lactate was
present in the medium. Since lactate consumption was
signicant only at the end of growth, it was concluded
that peptone was metabolized for cellular biosynthesis
and lactate for cell maintenance (137, 138). An increase
in pH in the curd then results from both the assimilation
of lactate as a carbon and energy source and the release
of ammonia through the metabolism of amino acids.
Ammonia is a signicant part of the aroma compounds
of Camembert (124). In the ripening of soft cheese,
ammonia in the atmosphere can reduce Penicillium
growth, thus lowering the level of proteolysis and bit-
terness (84).
The role played by G. candidum in controlling path-
ogens and contaminants in cheese continues to be
explored. The safety of raw milk mold-ripened soft
cheeses has been called into question due to the vulner-
ability of these cheese types to contamination, particu-
larly by Listeria monocytogenes, due to a high moisture
content and high pH of the rind (139). It has long
been assumed that pathogens will not survive in these
cheeses after a 60-day ripening period at a temperature
of 1.67C (>35F). However, recent studies and epi-
demiologic data have shown that neither a 60-day rip-
ening period nor, surprisingly, pasteurization ensures
the safety of this class of cheeses (140). Dieuleveux et al.
11
Marcellino and Benson

(141) have shown that G. candidum can inhibit the

growth of L. monocytogenes. The inhibitors were iden-
tied as D-3-phenyllactic acid and D-3-indollactic acid;
these compounds were stable over a wide pH range and
could be heated at 120C for 20 min. D-3-Phenyllactic
acid altered the cell wall of an antibiotic-resistant strain of
L. monocytogenes, producing depressions and holes that
were visualized with scanning electron microscopy (142,
241). G. candidum also inhibits the growth of the
zygomycete Mucor, whose aerial hyphae (cat hair) are
responsible for the cheese defect poil de chat (cat hair).
The inhibition is strain and culture dependent (143).
G. candidum may inhibit the sporulation of certain
strains of the mycotoxin producer Aspergillus avus
(144) and undesirable species of Penicillium such as P.
commune and P. caseifulvum, which can overtake fungal
starters (145). More research is needed on the potential of
G. candidum for microbial competition.
The results of our own studies of G. candidum sug-
gest that cheese technology and the microbial ecology of
a cheese rind select for populations of G. candidum
strains with different biochemical properties. The
variables in cheese technology and ripeningsalting,
humidity, temperature, and materials in the cave such as
wood or rye strawselected for strains with unique
characteristics. The fact that a strain from Bethlehem
cheese, a Saint-Nectaire-type cheese made in the United
States, would have biochemical properties similar to
those of strains from Saint-Nectaire in Auvergne,
France, tends to conrm the hypothesis that technology
breeds the strains.

The Price of Innumerable Failures: The Ugly

One will never know the patience and groping it took to put
into place the recipes for the fabrication of cheeses. It is the
principal merit of the humble farmers and silent monks to have
succeeded at the price of innumerable failures to rene and
perfect the assortment of cheeses of which we are justiably
very proud.
Les Fromages, Pierre Androut (54)
Ask anyone who has tried making cheese and he or she
will probably tell a tale of innumerable failures before
and sometimes after success is achieved. The natural
succession of microorganisms (and sometimes fauna) on
cheese during ripening has its benets and risks. A
novice afneur may wonder if the pile of dust he saw on
a cheese or shelf an hour earlier really had moved to
another location. This observation would mark his rst
encounter with cheese mites, the microscopic Arachnida
Acarus farris and Tyrophagus neiswanderi (146)
FIGURE 4 Challenges for the cheesemaker. (A) Scanning
electron micrograph of a cheese mite. Left unchecked, cheese
mites can lead to a 25% reduction in the weight of a ripened
cheese. Photograph by William R. McManus, generously pro-
vided and used with permission of William R. McManus and
Donald J. McMahon, Utah State University. (B) Spoilage of a
cheese by contamination and invasion of the rind by the fungal
genus Scopulariopsis. (C) Scanning electron micrograph of
the zygomycete Mucor showing sporangia among collapsed
hyphae. When the fragile outer membrane, the peridium
(arrowhead), of the sporangium breaks open, hundreds to
thousands of spores can be released, contaminating a cheese
cave overnight. Bar, 10 m. doi:10.1128/microbiolspec.CM-
0005-2012.f4.

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 The Good, the Bad, and the Ugly: Tales of Mold-Ripened Cheese
(Fig. 4A). In the Auvergne, cheesemakers call mites
artisans, and in Haute-Savoie, they call them cirons.
Cheesemakers may notice that mites are abundant after a
humid season, following heavy fungal growth on the
rind. They have asked, Are the mites eating the cheese or
the fungi? Tyrophagus mites isolated from the Spanish
mold-ripened cheese Cabrales are indeed fungivores
(147). Tyrophagus putrescentiae commonly infest
foods with high fat and protein contents (148), which
would certainly include cheese. The compounds cis- and
trans-octa-1,5-dien-3-ols produced by Trichothecium
roseum have been found to be strong attractants for the
mite (149). There is a relationship between mites and
fungi from which both may benet. Fungal spores can be
transported to new substrates, and mites can promote a
compensatory growth of fungal mycelium as new
surfaces are created for the aerobic fungi (150). The
mites choice of species of fungi upon which to feed
depends upon their capacity to digest the fungus. Cell
walls of fungal mycelium are composed of chitin and
chitosan, but in experiments on the digestibility of fungi
by various mite species, chitin passed through the di-
gestive tracts of mites undigested. Trehalose in the my-
celium, however, was digested by the mites, indicating
that the mites possess the digestive enzyme trehalase
(150). Of the mite species tested, trehalase activity was
highest in the Tyrophagus.
The presence of mites affects the appearance of the
rind and certainly the taste of the cheese. According to
Snchez-Ramos et al., Mites feed and reproduce on
cheese surfaces producing an accumulation of dead
mites, faeces, exuvia, eggs and bits of food, which ap-
pear as a light brown dust that can reach a depth of 2 cm
or more in heavy infestations (146). This may not
appeal to everyones tastes, yet Tyrophagus casei mites
are deliberately introduced into the German cheese
Altenburger (151). The very hard cheese Mimolette from
Lille, France, which can be aged for 2 years, develops its
distinctive avor with the help of mites. Yet on the whole
mites are a nuisance, especially for Cheddar makers, and
their removal is time-consuming, expensive, and often
frustrating.
At 25C and a relative humidity of 80 to 90%, one
generation of fungivore mites takes 10 days. Adult mites
can live for about 2 to 5 months, and Acarus farris, the
predominant species in Cabrales cheese, can reach a
population density of 260 mobile mites/cm2. It should
come as no surprise, then, that these artisans carving and
eating their way through the cheese surface, if left un-
checked in a cave, can be responsible for a loss of up to
25% by weight of the nal product (146).
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Not only do cheese mites cause economic loss, but
also their presence can cause enteritis, diarrhea, and
damage to the urinary tract (148). They are the source
of allergens implicated in asthma, rhinitis, and atopic
dermatitis. The storage mites Acarus siro, A. farris,
and Tyrophagus putrescentiae can cause occupational
allergies for farmers and cheesemakers (152), and the
afneur who spends long hours without a mask in a
mite-infested cave is vulnerable to a condition known as
cheesemakers lung caused by inhaling mites and their
dust (153). Mites can be the vectors of microorganisms
(154), and some have even suggested that hay mites may
be a self-sustaining reservoir for the prion which causes
bovine spongiform encephalopathy, or mad-cow disease
(155).
Control of mites continues to be a challenge for
cheesemakers. Generally, conditions in a cave which are
best for cheese ripening, with temperatures ranging from
10 to 15C and relative humidity near 90%, are also
favorable for mite populations (156). In the past, methyl
bromide fumigation and organophosphate insecticides
were used to control mites, but these have been banned in
consideration of food safety (156, 157). Many afneurs
still nd that mechanically removing mites from cheese
by brushing or vacuuming is the best and safest method.
Other measures have been tried, such as washing cheeses
with hydrogen peroxide after vacuuming and introduc-
ing ozone into the cave for a few hours a day (158). The
Bleu Mont Dairy Company in Wisconsin has used dia-
tomaceous earth (DE) as a successful material for mite
control. DE does not leave harmful residues, nor is it
toxic to mammals (159). DE injures mites through a
physical mode of action, rather than chemical. The mites
protective cuticle is damaged through abrasion and ad-
sorption of the cuticular waxes to the DE, which leads to
dehydration of the insects body (160).
Other approaches to control include the use of
modied-atmosphere (MA) technology that alters the
natural ratio of the atmospheric gases, oxygen (O2),
nitrogen (N2), and CO2 (157). Mobile stages of larvae,
nymphs, and adults are controlled quickly, whereas
preventing egg hatching is more difcult. One study
reported 100% mortality in the egg stage using 99.5%
CO2 at 85% relative humidity after 6 days of exposure,
while the mobile stages of the mites were killed in 1 day
under the same conditions. Preventing egg hatching
should therefore be the goal of any technology. Mono-
terpenes, including eucalyptol, showed acaricidal activ-
ity against mobile stages of Tyrophagus putrescentiae
through vapor action but adversely affected the avor
and odor of the treated cheese, so this approach was
13
Marcellino and Benson
abandoned (161). Other approaches include altered
ripening temperature and humidity levels; a lower rip-
ening temperature used with Cabrales cheese reduced
mite density (156).
Problems with fauna aside, some fungi are considered
contaminants that lead to defects in appearance and
avor (162) and, on occasion, the loss of entire batches of
cheese. The American mycologist Charles Thom de-
scribed the lamentous fungus Scopulariopsis brevicaulis
growing on Camembert in 1930 while working for the
U.S. Department of Agriculture at the Agricultural Ex-
periment Station in Storrs, CT. He described how
S. brevicaulis attacks Camembert cheese, overgrowing
the Camembert mold, producing an ammoniacal odor
and imparting an ammoniacal taste to the cheese (163).
In 1951 Keilling et al. described what they called a
cheese canker in Emmental, Gruyre, Edam, and
Gouda, which manifests itself during ripening and
storage through the appearance of little holes dug into
the rind of the cheese, containing a white, yellow, or
brown oury substance. They attributed the canker to
yeast and a fungus, Penicillium brevicaulis, a previous
name for Scopulariopsis brevicaulis (164). Unlike most
fungi, which prefer an acidic medium for growth,
Scopulariopsis grows best in alkaline and high-nitrogen
substrates, with an optimum growth pH of 9.0 (163).
Scopulariopsis reportedly is more apt to grow on pas-
teurized rather than raw milk Camembert and Carr de
lEst, because the lactic acid bacterium population in the
pasteurized version is less complex, leading to a weaker
fermentation and lower acidity (165). Scopulariopsis
spp. can contaminate many classes of cheese, with
Scopulariopsis fusca predominating, followed by S.
brevicaulis (108, 134). Samples of the aged pte molle
cheese Pont-lvque collected in four different de-
partments of Normandy showed contamination by the
fungus, as did aged artisanal goat cheeses and pressed
hard cheeses such as raclette and Gruyre (108, 134).
S. fusca appears as powdery brown or mauve spots on a
cheese rind and is a common contaminant along with
Penicillium species (108, 134). Artisanal cheesemakers
recognize only too well the cankers in the rind and the
strong smell of ammonia produced by Scopulariopsis in
their caves. The holes in the rind created by
Scopulariopsis allow it and other fungi to penetrate into
the curd. To add insult to injury, Keilling et al. warned
that mites invade the cavities which they extend con-
siderably deeper (164). Many cheesemakers have ob-
served that Scopulariopsis, besides destroying the
integrity of the rind as shown in Fig. 4B, creates defects in
avor such as bitterness and a musty taste.
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Scopulariopsis species are commonly found associat-
ed with soil, plants, and animal dwellings (166, 167).
They are considered anamorphs of the fungal class
Microascaceae (240) and microscopically can be con-
fused with Penicillium species. The conidia of
Scopulariopsis are oval and form in chains, with a
truncated base attached by a ring to a vial-shaped co-
nidiophore (165). The conidiospores often have rough
surfaces and are hyaline. Though not food-borne path-
ogens, ve species have been associated with human
diseases, and they can be serious emerging pathogens
for immunocompromised individuals (167, 168, 169).
The fungus possesses an array of enzymes that increase
its capacity to grow and ourish within the environment
of a ripened cheese rind. It is highly proteolytic: its
keratinases, which break down keratins, the principal
proteins in hooves, claws, and feathers, are being studied
for use in the eld of bioremediation (170). Its mode of
invasion of a substrate is specialized boring hyphae that
penetrate bers perpendicularly to the surface (171).
Considering that the fungus can penetrate the spines of
hedgehogs and cattle hair, it should not be surprising that
Scopulariopsis species bore their way through cheese
rinds. Since it prefers an alkaline environment, from an
ecological standpoint it follows that Scopulariopsis would
grow late in the ripening process. On a ripened cheese
surface, yeasts and fungi raise the pH through lactate
metabolism and ammonia production via proteolysis (44).
Bothast et al. noted that S. brevicaulis breaks down more
protein than it uses for synthesis when insufcient car-
bohydrate is present in the substrate (163), which is
probably the case in ripened cheeses, which contain only
trace amounts of residual carbohydrate (172). The fungus
then liberates nitrogen as ammonia from the excess and
utilizes carbon for energy production (163). Atmospheric
conditions in a cheese cave, including concentrations of
CO2, oxygen, and ammonia, determine microbial growth
and enzyme activity (173). In their study of atmospheric
conditions during ripening of Camembert, Leclercq-Perlat
et al. found that at high levels of CO2 and low levels of
oxygen, the yeast-like fungus Geotrichum candidum grew
well, whereas Penicillium roqueforti was negatively af-
fected (174). It would seem that an efcient ventilation
system in a cave which would allow for adjustments in
atmospheric conditions, particularly removal of ammo-
nia, would play a role in controlling Scopulariopsis. A
low-tech natural solution used by some cheesemakers to
slow down Scopulariopsis growth is to wipe the cheese
surface and shelves with vinegar.
Among its other enzymes, S. brevicaulis has an ex-
tracellular chitin deacetylase, which enables it to use
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 The Good, the Bad, and the Ugly: Tales of Mold-Ripened Cheese
chitin as a sole source of carbon (175). This enzyme may
enable S. brevicaulis to feed on fungi of preceding
populations on the cheese rind, using chitin contained in
their cell walls. Other enzymes of Scopulariopsis, such as
xylanases, are of interest to the paper and pulp industry
for their bleaching capacity and the breakdown of lignin
(176). Scopulariopsis is often carried into the ripening
environment on wrapping paper, so such paper should
be stored in a separate area (134, 177).
Scopulariopsis species are halotolerant, halophilic,
and xerophilic; they grow well in salty foods such as
cured sh and smoked bacon (178), in environments of
high salt concentration (179), and in salt-based media
(180). During cheese ripening, water evaporates from the
surface, thus lowering the water activity of the rind. Fox
et al. report that after 10 days of growth in nutrient broth
of pH 6.6 at 25C, Scopulariopsis fusca grew at 14% of
its maximum development at a salt concentration of
20% (water activity, 0.880). At the same time, Penicil-
lium candidum grew at 1.1% and Mucor mucedo and
Geotrichum candidum were completely inhibited (172).
Keilling et al. suggested that on Comt and Gruyre-type
cheeses, meticulous wiping and care of the rind are crit-
ical during the rst 15 days of ripening, when the cheese
rind is still acidic and the desired morge, or smear, is still
developing on the surface. When the morge does not
develop quickly enough, patches of S. brevicaulis can
appear (164). The members of the microbiota compris-
ing the morge, including Brevibacterium linens, are salt
tolerant and from the 15th to 30th day of ripening reach
a maximal growth of 109 to 1010 microorganisms per
cm3 on the rind (181). Adjusting salt concentrations and/
or care of the rind may inhibit undesirable fungi such as
Scopulariopsis and select for desirable species to compete
for nutrients on the cheese surface.
Considering what a widespread problem Scopulariopsis
is for the cheese industry and the lack of dairy-related
research data currently available, cheesemakers would
benet from investigations into modes of its inhibition.
In a recent investigation into antifungal activity by bacte-
ria, eight strains of lactobacilli isolated from vegetables and
one strain of Propionibacterium jensenii of dairy origin
strongly inhibited the growth of Scopulariopsis brevicaulis
(182). It is of interest to note that while many classes of
fungi were inhibited, Geotrichum candidum was not,
which is good news for cheesemakers. Testing of fermen-
tative bacteria that exhibit antimicrobial activity while not
affecting sensorial quality within a cheese environment
could be of interest.
The fungus that is probably the most common cause
of what French cheesemakers call accidents de fromages
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(loosely translated as cheese disasters) is the zygomycete
Mucor. As mentioned earlier, Mucor is part of the nor-
mal microbiota of Saint-Nectaire fermier (farmstead)
cheese and Tomme de Savoie (58, 63). However, when
Mucor appears on a pte molle cheese, it can instill fear
in the heart of a cheesemaker or afneur. A cheese such
as Camembert, meant to be covered with a uniform
white coat of Penicillium camemberti, can become cov-
ered with tufts of grayish-white hairs, a defect known as
poil de chat. Spore germination is the critical step in the
growth of food spoilage fungi because a product is
spoiled as soon as visible hyphae can be observed
(183). At the tip of each sporangiophore in Mucor is a
sporangium that appears as a black or gray ball to the
unaided eye (143). The electron micrograph of the sur-
face of a 9-day-old Bethlehem cheese in Fig. 4C shows
two sporangia among collapsed Mucor hyphae. One can
see the fragile outer membrane, called a peridium, of the
larger sporangium breaking apart, revealing the spores.
One sporangium can liberate thousands of spores (108)
that will spread over the cheese surface and eventually
impart a grayish coat to the cheese. This may explain
why Mucor has earned the name la bte noire (the black
beast) by some traditional cheesemakers. It grows 5 to
10 times faster than Penicillium species (108), giving it a
competitive advantage over Penicillium camemberti.
When Mucor racemosus was grown in potato dex-
trose agar medium at 25C, its hyphal extension rate,
based on the measurement of colony radial growth, was
13.3 mm per day (183).
Mucor is ubiquitous in the environment and, because
of its proteolytic and saccharolytic activities, is respon-
sible for decomposing fruit, meat, and bakery products
(184, 185). Though not a food-borne pathogen,
Mucor can cause mucormycosis, an infection of the
sinuses, brain, or lungs to which immunocompromised
populations are most vulnerable (186). Zygomycetes
have rarely been reported to produce mycotoxins (183).
Mucor racemosus is the most widespread cheese con-
taminant, but other Mucor species associated with cheese
are M. plumbeus, M. hiemalis, M. globosus, M. fuscus,
and M. mucedo (108). A characteristic that distinguishes
Mucor from other genera of the zygomycetes is its ca-
pacity for dimorphism (187), allowing it to adjust its
morphology to changing environmental conditions. In
an anaerobic environment and in the presence of a fer-
mentable sugar, this lamentous fungus can assume a
yeast-like morphology (185). In 1876, Louis Pasteur
described the dimorphic capability of Mucor racemosus,
which was implicated in the fungal contamination of
beer (184). Le Bars-Bailly et al. (108) noted that in a
15
Marcellino and Benson
cheesemaking facility, the lamentous morphology of
Mucor racemosus predominates on the aerated surface of
a cheese, whereas under a cheese, where less oxygen is
available, cells may assume a yeast-like morphology.
Zygomycetes grow poorly under conditions of low
water activity (183). With Mucors preference for high
humidity, the dairy environment presents many oppor-
tunities for contamination to which cheesemakers must
be alert. Its optimum growth conditions include a tem-
perature of 20 to 25C, a pH of 5.0 to 6.0, relative hu-
midity of 90 to 95%, and water activity of >0.95 (188).
The spores of Mucor are described as myxospores due
to the tendency of their cell walls to absorb moisture,
facilitating their adherence to various surfaces. The
spores stick to clothes, cheese molds, utensils, and the
cheesemakers hands, thrive in liquids such as whey and
stagnant water, and are dispersed in moist air (188, 189).
Once contamination has occurred, it is difcult to get
under control, so prevention is the rst goal.
During cheese production, newly formed cheeses are
extremely vulnerable to contamination during the
draining of whey. The high humidity in the draining
room and the high moisture content of the undrained
curd favor Mucor development. The temperature of the
room ideal for drainage (24 to 27C) is also ideal for
germination of Mucor spores (189). Whey contains
abundant lactose (about 70% of dry solids) (172), and
through its -galactosidase activity, Mucor is able to
break down lactose into glucose and galactose (190).
The presence of lactose in the whey allows for growth of
the yeast-like form of Mucor in the anaerobic environ-
ment of the interior of the curd (189). Acidication of
the curd in production enhances the expulsion of whey
and reduces drainage time. If cheese is made with milk
with a high fat content, good drainage may not occur
(188, 189). The use of fresh milk for cheesemaking
minimizes the time that psychotropic bacteria can break
down the casein in milk, providing polypeptides as a
substrate for Mucor (189). It is also recommended that
draining cheeses be covered to avoid airborne spores.
Tormo and Barral suggest that if Mucor growth is visible
on cheese 1 to 2 days after the curd is placed in the
forms, one should suspect that the milk used in pro-
duction was the original source of contamination. When
Mucor appears 3 to 4 days after draining and the entire
cheese is covered with hyphae, then the curd, cheese
forms, and utensils should be suspected; if growth
appears on only one side of the cheese, the atmosphere in
the room is the source (188).
Mucor can be controlled by fungi and bacteria in-
volved in cheese production and ripening. The sporula-
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tion of Mucor can be partially inhibited by good growth
of G. candidum on the rind of a pte molle cheese during
the early stages of ripening. This competitive inhibition is
strain dependent for G. candidum (144, 191). Le Bars-
Bailly et al. observed that since Mucor needs access to
nutritive medium to grow, a previously implanted thick,
uniform layer of G. candidum creates a physical barrier
between Mucor and the cheese substrate (108). As
mentioned earlier, tolerance to salt plays a large a role in
growth and survival in or on cheese. Cheeses of the pte
molle type, such as Camembert and Brie, generally are
salted within the range of 1.5 to 2.0% (wt/wt). The
cheesemaker may have to adjust the degree and method
of salting if a good implantation of G. candidum does not
occur. Lactobacillus coryniformis subsp. coryniformis
Si3 has displayed a broad spectrum of antifungal activity
against Mucor hiemalis as well as Aspergillus and Peni-
cillium species (192), and Brevibacterium linens has
shown antifungal activity via thiol production (143).
Using modied atmospheres for preventing fungal
growth and mycotoxin production to extend the shelf life
of some kinds of food was evaluated (183). Taniwaki et
al. found that the growth of Mucor plumbeus and other
fungi could be reduced in atmospheres containing 20 to
40% CO2 with 1 to 5% O2 (193). Tormo and Barral
report that Mucor is sensitive to ammonium and UV light
(188).
Penicillium roqueforti and Penicillium camemberti,
growing naturally or added as secondary starters, con-
tribute to the avor and unique characteristics of blue-
veined and pte molle cheeses, respectively (52, 91).
However, when they or other species of Penicillium grow
on the wrong type of cheese, they are considered
contaminants that can cause great economic loss (108).
P. roqueforti can contaminate Camembert and other
pte molle cheeses (194) as well as hard cheeses such as
Emmental and Parmesan (195), and P. caseifulvum can
cause yellow spots in blue-veined cheeses (196). Other
Penicillium speciesP. commune, P. solitum, P. palitans,
and P. crustosumare especially problematic because
they are closely related genetically to P. camemberti.
P. camemberti, the widely used starter in Camembert
and Brie, is considered a domesticated species derived
from P. commune (195). The taxon P. commune was rst
introduced by Thom in 1910 because it was the major
source of cheese contamination in the United States (197).
According to Lund et al. (198), it remains the most
widespread spoilage fungus, causing discoloration and
avor defects in cheese. The authors have observed that
one conidium of P. commune on a cheese can develop into
a colony of 10,000 CFU in 4 days. Therefore, great care
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 The Good, the Bad, and the Ugly: Tales of Mold-Ripened Cheese
must be taken to prevent the dispersal of contaminating
conidia in the air and on hands, clothes, and packaging.
Biocontrol of penicillia has also been noted. G. candidum
can inhibit the growth of P. roqueforti in blue-cheese
medium without NaCl, and some strains of P. camemberti
inhibit the growth of P. roqueforti, P. caseifulvum, and
P. commune (196).
TALES OF MOLD-RIPENED CHEESE
Difculties arise when one attempts to describe linkages
between natural and human history of cheesemaking in
the context of its science. Perspectives range from the
small farmer interested in terroirthat elusive property
associated with the specicity of space, including cli-
mate, soil, and peopleand the industrialist who wishes
to control all aspects of a ripening process to control
product consistency. An example of these differing
perspectives plays out in the Auvergne region of France.
The Auvergne is a volcanic region in the Massif
Central dominated by the Chane des Puys, a series of
volcanic cones, domes, and craters west of Clermont-
Ferrand (199). On the route from Montaigut-le-Blanc to
Champeix in Puy-de-Dme, the arched openings to the
caves of Saint-Julien where wine was aged before the
powdery mildew (Oidium tuckeri) and phylloxera
epidemics of grapevines, can be seen from a distance.
Some of those caves belong to the Bellonte family.
The elusive link of cheese to its environment is im-
mediately apparent as one enters the Bellonte caves and
sees hundreds of Saint-Nectaire cheeses laid out on rye
straw on the oor. The owner, Alphonse Bellonte, speaks
passionately about the patrimony of the Auvergne. He is
ercely attached to his land and cheese. The pride and
spirit of resistance expressed in his words are common to
the people of the Auvergne. The region is named for the
Celtic tribe Arverni, best known for their chieftain
Vercingtorix, who led his troops to victory over Julius
Caesars forces at Gergovia in 52 BCE (200). Although
he was later defeated and taken in chains to Rome, he
remains a symbol of heroism to the region (201).
Saint-Nectaire is an uncooked semihard surface
mold-ripened cheese that has been made since at least
the 17th century in two departments, southwestern Puy-
de-Dme and northern Cantal. It is a PDO cheese, and
the 69 communes of origin are characterized by the
geographical and geological diversity found in the Sancy
Mountains, the plateaus of Czallier and Artense, and
the valleys known as Pays coups (42, 202). The cheese
was named after Henri de Senecterre (15731662), who
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is said to have introduced it to the court of King Louis
XIV (203). The Bellonte family has made Saint-Nectaire
for eight generations in the town of Farges, 3 km from
Saint-Nectaire, a village located on the top of Mont
Cornadore and renowned for its Romanesque church
and ancient Roman thermal baths. Farges was built in
the volcanic crater of the Massif du Sancy, the oldest
mountain in the Auvergne, and is part of the Regional
Park of the Auvergne Volcanoes.
The decline in population and number of farms in
Farges since 1870 reects Alphonses sentiment: Mod-
ern agriculture has made French countryside a desert. In
1870, there were 90 inhabitants and 22 farms; in 1945,
46 inhabitants and 10 farms; in 1995, 23 inhabitants and
one 474-ha farm, that of the Bellonte family. During the
summer, cows graze in mountain pastures. The herd is
composed of Montbliardes, a breed originating in the
Jura region of France. In the past, milk of native breeds of
the Auvergne, such as Ferrandaise and Salers, was re-
quired for making Saint-Nectaire with the AOC desig-
nation (204). The Salers, a red cow whose ancestors were
depicted in cave art near the town of Salers in Cantal in
the Haute-Auvergne, is considered one of the oldest of all
European breeds (http://www.ansi.okstate.edu/breeds/
cattle/salers/index.htm). This dual-purpose cow is well
adapted to the rugged terrain and climate of the high
mountains. These cows are often pictured on labels for
Saint-Nectaire cheese (203), yet many farmers no longer
milk them because a calf must be tethered to the mother
during milking (205). This requirement makes for a
fairly chaotic pastoral scene not necessarily practical for
dairy operations today.
The Bellonte Montbliarde cows are fed only hay
and grain, never silage. Silage is not allowed in the pro-
duction of most AOC cheeses since it may contaminate
milk with spores from Clostridium species, particularly
C. tyrobutyricum and C. butyricum. These strains pro-
duce carbon dioxide and hydrogen gases during the bu-
tyric acid fermentation that cause openings, splitting, and
blowing defects in cheese, particularly in large cheeses
such as the Swiss types (206, 207). Silage can also be a
source of mycotoxin-producing fungi and the food-
borne pathogen Listeria monocytogenes when silage has
undergone deterioration aerobically (208, 209, 210).
The Bellonte farm makes Saint-Nectaire fermier cheese
using raw milk produced only on their farm. The produc-
tion of Saint-Nectaire laitier (industrial Saint-Nectaire)
using pasteurized milk collected from many producers be-
gan in the region in 1964, and at the time it was feared that
farmstead production would be threatened (211). How-
ever, Saint-Nectaire fermier is alive and well: of the 13,676
17
Marcellino and Benson
metric tons of Saint-Nectaire produced in 2005, 6,194
metric tons were fermier and 7,482 metric tons were laitier
(http://www.fromages-de-terroirs.com/fromage-detail.
php3?id_article=1634&lang=en). The Bellontes make 45
metric tons of Saint-Nectaire fermier annually.
The bulk of Saint-Nectaire produced by the Bellontes
is aged in the caves of Saint-Julien that were dug from
tuff (porous volcanic rock) in 1813; some also is aged in
caves in Farges dating to medieval times (742814 CE).
The medieval caves were dug from stone, called pierre de
Farges, that is very porous and retains moisture, thus
providing an excellent natural environment for the
growth of fungi. Until the 1890s, the caves of Saint-Julien
were used to store wine, but by then the effects of phyl-
loxera (Daktulosphaira vitifoliae), the grapevine louse
that destroyed the vines of France, had reached the
Auvergne. As Gale describes it, the entire wine-
growing world was not only affected but transformed
agriculturally, economically and sociallyby the plague.
Effects of the disaster rippled out from the vineyards into
their embedding cultures, invoking such large-scale
consequences as rural depopulation and massive
emigration (212). Until that time, the Auvergne had
been the third-largest winemaking region of France, but
it would never again regain that stature.
Many of the caves in the countryside of Saint-Julien
are abandoned, but others are used to store root vege-
tables or ripen cheese. In compliance with regulations of
the European Union, the cheeses are now ripened on
plastic mats on wooden shelves and not directly on rye
straw on the cave oor (213). In many Saint-Nectaire
caves, the cheesemaker slips stalks of rye straw between
the mats and shelves or into the plastic crates of aging
cheeses. This modication may be seen as resisting
the regulations, or as Vollet et al. suggest, the cheese
is presented on a bed of rye straw as a marketing tool
for tourists, to accent terroir (203). However, rye straw,
or paille de seigle, cannot be viewed apart from the
Celtic roots of the Auvergne and the history of the cheese.
The name Czallier has a Celtic origin; the French
term for the region is pays du seigle, or country of rye
(214). Rye grain was used to make bread in communal
ovens in mountain hamlets and rye straw was placed
under the ripening cheeses. As Greliche has noted, Rye
straw was so important that it gave its name to the
cheese that was produced locally called le gleo which is
considered as the ancestor of St. Nectaire. The Celtic
words gleo, glahou, and glui signify the rye straw on
which the cheese was ripened (213). During the Middle
Ages, peasants paid their lords glo cheese as a form
of currency.
18
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The cheesemakers of the Auvergne have observed for
centuries that rye straw placed under ripening Saint-
Nectaire seems to produce the best cheeses. From a mi-
crobiological standpoint, the use of rye straw is a factor
in fungal succession on the cheese rind: namely, it
encourages the growth of Trichothecium roseum, the
ower of the molds. T. roseum is a saprophyte as well
as a plant pathogen (215). The fungus is associated with
wheat, especially during seasons of high precipitation
(216), and is among the fungi that can use wheat gluten
as a substrate in the fermentation of minchin, a Chinese
cereal-based food (2). T. roseum can overwinter in soil
or crop debris, while under conditions of high humidity,
such as one would nd in a cheese cave, it grows well
and sporulates (217). Proteases of T. roseum may war-
rant investigation to clarify the role of T. roseum in the
microbial ecology of cheese ripening.
Some French cheeses have emerged bearing a mix of
natural and political history that includes universal
truths about the struggles and ingenuity of small farms
and partly explains the passion with which traditions are
held. Reblochon, for example, is a cheese that originated
in the 14th century in the Haute-Savoie region in the
Chane des Aravis, a mountain range of the pre-Alps
(218). It received its AOC appellation in 1958 and is
among the rst French cheeses to do so. The story behind
its inception is often recounted with pride by the French.
Reblochon is derived from blocher, which in Savoyard
dialect literally means to pinch a cows teat. According
to the peasants hand-milking a cow, the sound of a
stream of milk hitting a pail is similar to ploutch, which
developed into the verb blocher (http://projetbabel.org/
forum/viewtopic.php?t=9246). Re-blocher means to
milk again, and a cow which holds back her milk and
needs to be milked a second time is called une vache
reblochnieuse (219). In the 13th century, farmers were
taxed according to the quantity of milk produced by their
herds, so they began the practice of not milking the cows
out completely on the day of the tax collectors visit.
After the tax collector departed, the cows were milked
again. To protect the ruse, the small quantity of rich milk
obtained from the second milking was quickly
transformed into cheese. There is a play on words in
Savoyard between reblocher (to milk again) and rablasse
or rablache (a fraudulent action), and thus Reblochon
came to be known as the cheese of fraud or deception
(220). Fresh raw whole milk is still required for pro-
ducing AOC Reblochon today, and the cheese is made
immediately after milking, twice a day. Reblochon
fermier (with a green identication label) is made on
a farm with milk produced on that farm only,
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 The Good, the Bad, and the Ugly: Tales of Mold-Ripened Cheese
whereas Reblochon laitier (with a red label) is made in a
dairy from milk collected from many farms. In both
cases, the cheese is made from milk of three breeds:
Abondance and Tarentaise, both native to Haute-Savoie,
and Montbliarde. Reblochon is considered a semihard
cheese, but Mariani (75) notes that it could be classi-
ed technologically as a transition between pte molle
(moist) cheese, to which it is similar at the beginning of
ripening, and a pressed cheese at the end of ripening.
It is washed twice during the 4- to 6-week ripening
period, encouraging the growth of the coryneform
Brevibacterium linens and Geotrichum candidum, which
is the only lamentous fungus on Reblochon (221).
On Reblochon, Mucor is considered a contaminant.
Brtschi et al. (221) suggested that contamination of
Reblochon with Mucor may be less common than in
pte molle types because the curd is pressed, thus
shortening the draining period, when the cheeses are
most vulnerable to contamination. Washing the rind
during ripening may also reduce the risk. However,
Reblochon is not immune to contamination, and some
stories illustrate the vigor with which Mucor can foment
disaster. One cheesemaker in Haute-Savoie recounted a
time in the 1950s when on a Wednesday she returned
from her cheese cellar and exclaimed to her husband,
How beautiful the Reblochons are! The ripened
Reblochons were ready to go to market on Saturday. But
by Friday morning, the cheeses were covered with poil
de chat (Mucor), and in the evening, they were com-
pletely covered with black sporangia. The dairy con-
sultant from La Roche-sur-Foron did not nd Mucor in
their milk and concluded that such a drastic contami-
nation could only be due to a mauvais sortsomeone
had put a spell on their cheese. He suggested that they
contact the priest to perform an exorcism. The outcome
of the exorcism is unknown. The competition between
Mucor and Geotrichum can be unpredictable and may
be one-sided in new caves that have not had time to
establish appropriate microbial populations. For that
reason, newly established Reblochon caves are some-
times watered down and seeded with Geotrichum
candidum (75).
In the area of the Haut-Doubs in the region of
Franche-Comt, the Sancey-Richard family has been
making cheese since 1961. At 1,463 m of altitude, the
Fromagrie du Mont dOr of Mtabief is located 10 km
from the Swiss border at the base of Mont dOr, a summit
in the rugged Jura Mountain range. The Jura Mountains
traverse the Franco-Swiss border north of the Alps be-
tween the Rhne and Rhine rivers and were named by the
Celtic tribe the Sequani, who inhabited them in the 4th
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century BCE (222). The Celtic root jor, later Latinized to
juria, means forest, which aptly describes these forest
mountains dominated by forests of Norwegian spruce
(223). It was here near the source of the Doubs River that
the cheese which perhaps best expresses the landscape
from which it arose, Mont dOr, was created. There are
two versions: the French-called Mont dOr or Vacherin
du Haut-Doubs, which is made from raw milk, and the
Swiss version, Vacherin Mont dOr, made from
thermized milk. Mont dOr is a seasonal cheese which
according to French law can be made only from 15 Au-
gust to 31 March (224). Cheesemonger and writer
Patricia Michelson describes well the seasonal quality of
the cheese: The rst true taste of autumn for me comes
when the cheese table in my shop displays Vacherin
Mont dOr. Not for me the bland taste of early Septem-
ber cheeses: I prefer to wait until the end of Octoberor
the weekend when we turn back the clocks in Britain
until the most seductive and sensual of cheeses is avail-
able (225). The timing has its roots in the ancient
rhythms of mountainous regions. In late spring, the fte
de la transhumance or lestive, the day the herds and
ocks pass through the villages on their way to the higher
elevations for the summer, is celebrated with music and
dance (226). Traditionally, the herds came down the
mountains just after 15 August. A cow decorated with a
wreath of owers on her head leading a herd down the
mountain may be the cow that has given the most milk
that summer. Once the cows returned to their winter
quarters and were no longer grazing on lush pastures,
they gave less milk, and thus, Mont dOr, a small cheese
in comparison to other celebrated cheeses of Franche-
Comt such as Gruyre de Comt and Morbier, was
created. It is often called the Christmas cheese because it
is available during the holiday season, at which time it is
in great demand throughout France.
Mont dOr is a creamy crote eurie (owered
crust) cheese that must be made at an altitude of 700 to
1,400 m. At Mtabief, the cheese is produced with the
milk of local herds of Montbliardes, the popular breed
that has been exported from her native Franche-Comt
to many regions in France. The cheese is made in copper
vats, and Patrick Sancey-Richard explained that since
milk contains very little copper, the vats encourage the
growth of microorganisms that require copper for their
growth.
This master cheesemakers observation about the use
of traditional materials provides an example of the em-
pirical knowledge of microbial ecology gleaned through
centuries of hands-on experience, the complexity of
which scientic research is only beginning to elucidate.
19
Marcellino and Benson
European cheesemakers have long considered copper
vats essential to Emmental production. Sieber et al.
speculate that in the evolution of humans use of metal,
the extraction of copper from ores brought an end to the
Stone Age 9,000 years ago. Copper is a prized metal
because it is malleable and, second to silver, the best
conductor of electricity and heat (227). Copper is re-
quired as a micronutrient to serve as a cofactor for
metalloproteins and enzymes involved in microbial me-
tabolism (228, 229). Cow milk contains very little copper
(0.1 to 0.2 ppm) (228). However, the use of copper vats
for cheesemaking brings the copper content of the n-
ished cheese to 7.6 and 16.5 ppm due to the leaching of
copper ions by the casein in milk (230). As copper vats
have been replaced by stainless steel in the production of
Emmental, the addition of CuSO4 salt to the cheese milk
has become standard practice. However, in organic
cheesemaking operations, the supplementation with
copper sulfate is not allowed (231). Likewise, the con-
centration of added copper must be nely tuned, as some
authors have noted that supplementary copper can in-
hibit the growth of starter and adjunct cultures such as
Streptococcus, Lactobacillus, and Propionibacterium
species critical to Emmental production (230). Inves-
tigations into the inhibition of food-borne micro-
organisms by copper are promising. Mato Rodriguez
and Alatossava (231) report that the use of copper vats in
cheese production inhibits the germination, vegetative
growth, and sporulation of strains of Clostridium
tyrobutyricum, the bacterium known to cause the late-
blowing defect in Swiss-type cheeses. Noyce et al. (232)
found that food surfaces composed of a copper alloy
inhibited the growth of the food-borne pathogen
Escherichia coli O157:H7, whereas stainless steel and
aluminum surfaces did not. When they inoculated seven
different types of cast copper alloys with 103 CFU of
E. coli O157:H7, all of the cells were killed in 20 min or
less at 22C. Likewise, the food-borne pathogens Sal-
monella enterica and Campylobacter jejuni were in-
hibited by contact with copper surfaces, while surfaces
comprised of stainless steel and a synthetic polymer
showed no antibacterial activity (229). This research
underscores the complexity of microbial inhibition and
may cause us to reconsider advances in food safety
such as the use of stainless steel in certain food produc-
tion environments.
The use of natural materials can be required for
producing some PDO cheeses. The 2006 regulations for
Mont dOr production stipulate that the cheese must be
surrounded by a strip of spruce bark and placed in a
spruce box (75). The inner layer of spruce bark is cut
20
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into strips, or sangles, by an artisan called a sanglier. The
sangles are rolled into packets of 10 that are dried,
soaked in salt water for 24 to 48 h, boiled for 2 h, and
stored in salt water until use. After the cheese is removed
from the mold, a sangle is placed around the fragile
newly formed cheese to hold it together and secured with
a pine toothpick. During ripening, Penicillium candidum
grows on the bark, presumably because Penicillium is
salt tolerant. The use of salt in preparing the bark may
have selected for Penicillium in years gone by, although
most producers now add a commercial strain. Early in
ripening, G. candidum grows on the cheese surface. G.
candidum and the coryneform bacteria B. linens and
Arthrobacter create a whitish-orange crust characteristic
of Mont dOr (66). At 21 days of ripening, the cheese,
still encircled by the strip of bark, is placed in the spruce
box. The cheese is served in the box, and its creamy pale
yellow interior is often eaten with a spoon. The aroma is
dominated by the smoky avor of terpenes, compounds
contributed by the wood. Among the antimicrobial
terpenes isolated from Mont dOr are p-cymene, limo-
nene, -terpinene, borneol, and terpineol (75).
Several types of cheeses owe their origins, continu-
ance, or development to various abbeys in Europe.
These include common varieties such as Munster (from
the Latin Monasterium), Cheddar, Saint-Nectaire, and
Maroilles. St. Benedict, the founder of Western monas-
ticism (ca. 500 CE) describes the enclosure of the mon-
astery as a workshop in his Rule for monks (233). His
emphasis on manual labor as well as the stability and
rhythm of monastic life created an environment for de-
veloping knowledge based on experimentation. An ex-
ample is the creation of Maroilles cheese, which is
attributed to the Benedictine Abbey of Maroilles in
northern France in 960. A monument in the town bears
the inscription Stile created to commemorate the crea-
tion of Maroilles by the monks of the celebrated Abbey:
9601960. The cheese was rst called craquegnon but
was known in the Middle Ages as the marvel of
Maroilles (234).
The Order of Cistercians of the Strict Observance was
founded in the 11th century and as the name implies was
a reform movement within the Benedictine tradition.
A subsequent even stricter reform movement of the
Cistercian Order took place at the abbey La Grande
Trappe in Normandy in 1664, and thus the monks were
called Trappists. The Cistercians were known for their
technological innovation and for transforming wilder-
ness and malaria-ridden marshy land into farmland. The
monks reintroduced water wheels to Europe, technology
which had been abandoned by the Romans (235). For
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 The Good, the Bad, and the Ugly: Tales of Mold-Ripened Cheese
the sustenance of the monastic community, pilgrims, and
surrounding population, the monks developed food
technologies, including beer brewing and cheesemaking
(236). Cheese was an important part of the monastic diet
because according to Benedicts Rule, eating the esh of
quadrupeds was forbidden except by the inrm (237).
Since monastic meals were eaten in strict silence, the
monks used sign language to communicate serving
needs. The Code of Signs found in records from the
abbey of St. Thomas the Martyr in Dublin, founded in
1177, provides us with evidence that there were
cheesemakers at the monastic table:
Pro signo casei, utramque manum con junge per obliquum,
quasi qui caseum premit.
[For the sign of cheese, join both hands obliquely, as one who
presses cheese.] (238)
Tami cheese is made at the Trappist Abbaye Notre-
Dame de Tami, founded in 1132 in Haute-Savoie in the
Alps at 900 m. Since 1863, the monks have made the
cheese we now know as Tami, but presumably other
varieties were made far into the past before the aban-
donment of the abbey during the French Revolution in
1792 (239). From the archives of 1600 we read that
someone lost the key to the cheese cave.
Tami cheese was trademarked in 1946 and is made
and sold as a means of sustainability for the monastic
community, yet its production involves a social obliga-
tion on the part of the monks. In 1132, the abbey was
founded to provide food for the peasants of the area as
well as travelers passing through the Alps from Geneva
to the Little Saint Bernard Pass. During the 12th century,
the monks helped the peasants clear land for pastures
and roads and aided them in organizing agricultural
granges for large-scale cheese production (239). The
obligation to local farmers is continued today; the
monks use milk from 11 local farms for the production
of Tami. Clary (239) has noted that the supplier-pro-
ducer relationship between the monks and farmers is
maintained through professional contracts yet is built
upon hundreds of years of establishing trust. Many of
these farmers are descendants of those who maintained
the abbey property during and after the French Revo-
lution, and some are descendants of the original farmers
whose land was surrendered for the building of the
original abbey.
Frre Nathanal, the monk in charge of cheese pro-
duction, attended dairy school and worked in the in-
dustry for 9 years before entering monastic life. He calls
cheesemaking a complex alchemy and brings his
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professional and monastic training to cheesemaking.
The monks keep their artisanal cheese local by using
milk from breeds native to the Alps, the Tarentaise and
Abondance, which, when not grazing in alpine pastures
in the summer, eat feed from the region. Frre Nathanal
extends safeguarding the patrimony of the region to the
microbiota. Like other master cheesemakers, Frre
Nathanal wants to make a consistently uniform and
safe product yet does not want to lose the biodiversity of
local microorganisms. Studies have shown that com-
mercial cultures of starter bacteria such as Streptococcus
thermophilus and Lactococcus lactis quickly outnumber
the wild-type species initially present in raw milk (42).
Thus, in collaboration with a commercial culture com-
pany, he isolated bacteria from hay and milk of the re-
gion and developed a collection of starter cultures that
could be alternated in Tami production in case the
starters became infected by bacteriophage. Tami thus is
a cheese that stands at the crossroads of scientic and
traditional methods of cheese fabrication.
Frre Nathanals latest innovation involves the
recycling of whey, a by-product of cheesemaking and a
potential pollutant. Cheese production at the abbey
produces 1,000 m3 of whey annually, and Frre
Nathanal developed and installed a methane-generat-
ing system that transforms the whey into enough energy
to heat water for the domestic needs of the abbey and its
dairy for a year. The Abbey of Tami was awarded the
Rhne-Alpes 2005 Energies of Today prize.
The cheese cave of the abbey, with its arched vaults
and limestone walls, lies beneath the Monastic Choir.
Standing in the cave, Frre Nathanal shared the secret
of knowing when a Tami cheese is ready to be eaten.
One gently taps the surface, and if there is no indentation,
it is ready. One then smells it underneath. If a sent la
vache (it smells like the cow), it is good. In another
context, he compared the smell to a less acceptable by-
product of the cow. Everyone may not be at ease with
Frre Nathanals analogy, yet it reminds us of the
earthiness associated with the origins of cheese. In com-
paring the language used to describe wine consisting
generally of associations with various fruit and owers
to that of cheese, cheesemaker and philosopher Jim
Stillwaggon mused, Cheese on the other hand is less
discreet. If we address frankly what is evoked by cheese I
think it becomes clear why so little is saidusually little
more than sounds of approbation, and global adjectives:
Mmmmmm, good! Interesting! Fantastic! So what does
cheese evoke? Damp dark cellars, molds, mildews and
mushrooms galore, dirty laundry and high school locker
rooms, digestive processes and visceral fermentations,
21
Marcellino and Benson
he-goats which do not remind of Chanel . In sum, cheese
reminds of dubious, even unsavory places, both in nature
and within our own organisms. And yet we love it
(personal communication).
ACKNOWLEDGMENT
The authors declare that they have no commercial afliations,
consultancies, stock or equity interests, or patent-licensing
arrangements that could be considered to pose a conict of in-
terest regarding the manuscript.
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