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Traditional foods and beverages from South America: Microbial communities and production strategies

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Abstract

Spontaneous fermentation has been used for several thousands of years as an affordable source to preserve and enhance the quality of foods and beverages. Many fermented products are prepared in different parts of South America using raw materials such as milk, cassava, maize, cacao, coffee, grape, sugar cane, banana, and others. These traditional products can be used to make cachaça, wine, pisco, sour cassava starch, cheese, coffee, chocolate, vinegar and chichas which are important beverages and fermented foods consumed in South America. These fermented products have peculiar nutritional and sensory properties that derive from the biochemical transformations of specific raw materials. The diversity of the products varies according to the geographical area, cultural preference, fermentation techniques, local customs, and religious beliefs. Most traditional fermented foods and beverages of South America are produced on a minor scale using traditional recipes. In general, fermentations are conducted by complex microbial communities with the occurrence of yeasts, lactic and acetic acid bacteria and molds. Different microorganism species developed during fermentation, and their dynamics and frequencies of appearance determine a big fraction of the taste and flavour characteristics of these traditional foods and beverages. However, the microbial ecology of the fermentation processes is poorly studied in South America, and only in the last years several studies dealing with this subject are being published. The knowledge about the microbial ecology of food and beverage ecosystems is essential to understand the production process. In this chapter, we will discuss the production technologies and the microorganisms involved in the fermentation processes of several traditional foods and beverages produced in South America.
In: Industrial Fermentation: … ISBN: 978-1-60876-550-8
Editors: Jürgen Krause and Oswald Fleischer ©2009 Nova Science Publishers, Inc.
Chapter 3
TRADITIONAL FOODS AND BEVERAGES FROM
SOUTH AMERICA: MICROBIAL COMMUNITIES AND
PRODUCTION STRATEGIES
Fátima C. O. Gomes1, Inayara C. A. Lacerda2, Diego Libkind3,
Christian A. Lopes4, Javier Carvajal5 and Carlos A. Rosa2
1Centro Federal de Educação Tecnológica de Minas Gerais, MG, Brazil
2Universidade Federal de Minas Gerais, MG, Brazil
3Environment and Biodiversity Research Institute (INIBIOMA), Bariloche, Argentina
4Instituto Multidisciplinario de Investigación y Desarrollo de la Patagonia Norte (IDEPA,
CONICET-Universidad Nacional del Comahue), Neuquén, Argentina
5Pontificia Universidad Católica del Ecuador, Quito, Ecuador
SUMMARY
Spontaneous fermentation has been used for several thousands of years as an
affordable source to preserve and enhance the quality of foods and beverages. Many
fermented products are prepared in different parts of South America using raw materials
such as milk, cassava, maize, cacao, coffee, grape, sugar cane, banana, and others. These
traditional products can be used to make cachaça, wine, pisco, sour cassava starch,
cheese, coffee, chocolate, vinegar and chichas which are important beverages and
fermented foods consumed in South America. These fermented products have peculiar
nutritional and sensory properties that derive from the biochemical transformations of
specific raw materials. The diversity of the products varies according to the geographical
area, cultural preference, fermentation techniques, local customs, and religious beliefs.
Most traditional fermented foods and beverages of South America are produced on a
minor scale using traditional recipes. In general, fermentations are conducted by complex
microbial communities with the occurrence of yeasts, lactic and acetic acid bacteria and
molds. Different microorganism species developed during fermentation, and their
dynamics and frequencies of appearance determine a big fraction of the taste and flavour
characteristics of these traditional foods and beverages. However, the microbial ecology
of the fermentation processes is poorly studied in South America, and only in the last
Fátima C. O. Gomes, Inayara C. A. Lacerda, Diego Libkind, et al.
years several studies dealing with this subject are being published. The knowledge about
the microbial ecology of food and beverage ecosystems is essential to understand the
production process. In this chapter, we will discuss the production technologies and the
microorganisms involved in the fermentation processes of several traditional foods and
beverages produced in South America.
1. GENERAL CONSIDERATIONS
The production of fermented foods and beverages is one of the oldest technologies of
food processing kwon by humanity. Since the first civilizations methods for fermentation of
milk, meat and vegetables have been described, and the first records are dated of 6,000 BC
(Fox, 1999). These processes occurred by spontaneous fermentation without any
knowledgement on the role of the microorganisms. Scientific advances in the century XIX
showed the role of the microorganisms during the fermentations, and at the end of this
century, several starter cultures for different fermented foods were isolated, and these cultures
could be used for large scale production (Caplice and Fitzgerald, 1999). According to Aidoo
et al. (2005), traditional or indigenous fermented foods are those popular products that since
early history have formed an integral part of the diet and that can be prepared in the
household or in cottage industry using relatively simple techniques and equipment.
The beverages and foods traditionally fermented from South America are products
obtained by the modification of different raw materials from vegetables or animals, with the
active participation of microorganisms or enzymes, improving the flavor of these products.
Yeasts are involved in the production of several kinds of traditional fermented foods and
beverages (Haard et al., 1999; Jespersen, 2003; Aidoo et al., 2006). Lactic acid bacteria are
widely used for the preparation of fermented foods, such as foods based on starch (cassava,
maize and rice) and dairy products, which are of great importance for the countries of South
America. There is a wide diversity of lactic acid bacteria and yeasts within a variety of food-
related ecosystems (Damelin et al., 1995; Lacerda et al. 2005), and different microbial
populations are responsible for the fermentation of these substrates with the formation of
several fermented products.
Food fermentation has historical, philosophical, archaeological and religious significance
(Echeverría and Muñoz, 1988; Steinkraus, 1997; Ray et al., 2009). Most of the fermented
foods have evolved with time and are food substrates that are invaded or overgrown by edible
micro-organisms whose enzymes, particularly amylases, proteases and lipases hydrolyses the
polysaccharides, proteins and lipids to products with flavours, aromas and textures pleasant
and attractive to the human consumers (Ray and Sivakumar, 2009). Fermentation plays at
least five roles in food processing: enrichment of human dietary through development of a
wide diversity of flavours, aromas and textures in foods; preservation of substantial amounts
of foods through lactic acid, alcoholic, acetic acid, alkaline and high salt fermentation;
enrichment of food substrates biologically with vitamins, proteins, essential amino acids and
fatty acids; detoxification during food fermentation processing and decrease in cooking times
and fuel requirements (Steinkraus, 2002).
A vast range of traditional fermented products are produced in South America, where,
along with lactic acid bacteria, a diversity of yeast species makes important contributions.
Lactobacillus plantarum and other lactic acid bacteria and yeasts have been shown to be
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Traditional Foods and Beverages from South America: …
prevalent microorganisms associated with cassava fermentation (Figueroa et al., 1995;
Carvalho et al., 1999, ben Omar et al., 2000; Ampe et al., 2001, Lacerda et al., 2005). Coffee
beans and cocoa beans (chocolate) undergo natural, indigenous fermentation during the
primary stages of their processing, at which time the growth and activities of a diversity of
Hanseniaspora, Candida, Pichia Issatchenkia, Kluyveromyces, and Saccharomyces species
have been reported. Essentially, these yeasts assist in the degradation of bean pulp and
contribute to the production of chocolate flavor precursors (Schwan and Wheals, 2003, 2004).
Several traditional cheeses are produced in South America, and lactic acid bacteria, primarily
Lactococcus lactis, are responsible for the fermentation process while yeasts influence the
aging process (Borelli et al., 2006). Grapes have been used as the main raw material in the
production of wine. However, a number of researchers have identified other suitable fruits for
wine production. Over the years, fruit wine has been prepared from several different fruits,
such as cajá, banana, pupunha, mango, acerola, and cocoa (Whasley et al., 2009). Sugarcane
juice is used to prepare cachaça, the most traditional Brazilian distilled beverage. This
beverage is mostly produced by spontaneous fermentation, in which Saccharomyces
cerevisiae is the prevalent species (Gomes et al. 2007). Chicha is a beverage consumed by
Andean indigenous populations (Hastorf and Johannessen, 1993), and its flavor is similar to
that of cider. Champús, masato, cauim, and traditional beers are other popular beverages in
some countries of South America. In this chapter, we will provide an overview of these
traditional products, showing some aspects of the technological process and of the microbial
communities involved in the fermentation of these beverages.
2. TRADITIONAL BEVERAGES FROM SOUTH AMERICA
Traditional beverages from South America have been used as stimulants in traditional
medicine as well in religious ceremonies for many years. However, the introduction of
industrial beverages has decreased the use of these traditional products (Wacher-Rodarte,
1995). A great diversity of traditional beverages is produced in South America, but studies on
the microbial populations associated with the fermentative processes of these products are
rare.
The beverages possess peculiar nutritional and sensory properties derived from the
fermentation of specific raw materials (Campbell-Platt, 1994). The preparation of many
indigenous or traditional fermented beverages is still an art in the home, in villages, and in
small-scale industries. In general, natural fermentation is carried out by yeasts, lactic acid
bacteria, and fungi, sometimes forming complex microbiota that function cooperatively
(Blandino et al., 2003). Several traditional beverages will be discussed below, such as Chicha,
a starch-fermented beverage; masato, a beverage obtained from cassava roots; beer; cachaça,
the most popular Brazilian distilled beverage, produced from fresh sugar cane juice; champus,
a popular cereal-based, low alcoholic beverage; caiuins, a beverage produced from cassava
fermentation by Brazilian Indians; and the wine produced in some regions of South America.
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Fátima C. O. Gomes, Inayara C. A. Lacerda, Diego Libkind, et al.
2.1. Chicha
Chicha, or corn beer, is the most important fermented beverage studied in South America.
Chicha is a clear, yellowish, sparkling beverage that was consumed by indigenous Andean
populations for hundreds, maybe even thousands, of years (Hastorf and Johannessen, 1993;
Jennings et al., 2005). Its flavor resembles that of cider. The traditional production process of
chicha is unique, because the amylase from saliva is used to convert starch into fermentable
sugars.
Chicha is a generic name that encompasses a wide variety of fermented alcoholic
beverages. These beverages are made from different kind of raw materials, most commonly
from corn. The sugar source for the fermentation process is sometimes obtained from other
vegetables, such as cassava, mishqui (the sweet juice from Agave americana), cane sugar, and
a wide variety of mixtures, including rice, oat flour, or corn starch. It is believed that the
name of this beverage comes from the word “chichab”, from the original language spoken in
the current Panama territory and meaning corn (Diccionario de la Real Academia Española de
la Lengua on line http://buscon.rae.es/draeI/SrvltConsulta?TIPO_BUS=3andLEMA=chicha).
Others believe that this word could also be a deformation of the name “Chibcha”, a
civilization that settled in Colombia and Panama.
Prior to the Inca’s conquest, an etnia existed in Southern Bolivia called the “Chichas”.
Currently, the Nor Chicha and Sud Chicha provinces in Bolivia resemble the name of the
ancient etnia (E. C. Arispe, personal communication). In any case, the word chicha in the
Ecuadorian territory was introduced by Spanish conquerors. The local word used for this
beverage, after the Inca invasion in the current Ecuadorian territories was asua, from the
Quichua language (M. C Molestina, personal communication). Archaeological findings
indicative of chicha production in a number of Andean locations are very common, and often
the findings are discovered in areas where administrative centers of Andean peoples existed
(Hayashida, 2008).
The first register of chicha in Ecuador is probably from the year 200 A.C., when the
Hipia people occupied the territory between the Quito and Nariño region in Colombia. This
archaeological finding demonstrates that chicha was being produced long before the Incas
invaded the territories of the current Republic of Ecuador (Molestina, 2006). The plateau of
Quito was a commercial exchanging zone used by different human groups from a wide range
of geographical regions. Archaeological evidence demonstrates that Quito was already
inhabited in 100 A.C., and probably even earlier (S. Moreno, personal communication).
Later, during the Inca conquests, some etnias originating from distant regions of the Inca
Empire (Tahuantinsuyo) were implanted, providing a mixture of costumes and different
techniques for the production of chicha. Finally, Spaniards arrived along with their culture
and fermentation technology around 1534 (Echeverría and Muñoz, 1988). Thus, Quito
constituted a center in which different fermentation technologies and different fermenting
yeast species converged and coexisted throughout history (S. Moreno, personal
communication)
2.1.1. Manufacturing
Archaeological findings show that approximately 1800 to 1360 years ago, the Andean
people from the “Hipia” culture settled in the slopes of the Pichincha volcano, close to Quito,
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Traditional Foods and Beverages from South America: …
Ecuador. These people discovered that germinated grains of corn are suitable sources of sugar
for the production of sweet broths which constitute the basis of the stimulating beverages.
Most likely by accident, corn kernels were germinated and then dried by the sun. In this way,
germination was stopped as the endosperm of the kernels was modified by enzymes produced
during germination. The result of this germination and drying processes was the so-called
jora”, or corn malt. Jora became the most important raw material for the manufacturing of
chicha due to its ability to produce sweet worts after a period of boiling.
The whole cooking procedure was carried out in vessels made out of clay and probably
“cured” with cornstarch. The curing of vessels was very important for the maintenance of the
liquid contents of the receptacle; otherwise, if not cured correctly pores in the material could
lead to permeability, eliminating liquid retention. Thus, curing was a way to cap pores using a
material that, once hydrated and cooked, became sticky (M. C. Molestina, personal
communication).
Fermentation took place inside a special kind of clay vessel called a pondo”. This
fermenter has a particular shape, with a restricted mouth to avoid contamination of the
ferment ting material inside and an acute-shaped bottom for placement within a hole
excavated in the soil. Thus, the pondo is protected from the outer temperature, thus
maintaining stable thermal conditions during the fermentation process (S. Moreno, personal
communication).
There are a number of different ways to make chicha. As this is not a written technology
but the tradition has been passed from one generation to the next for hundreds, potentially
even thousands of years, many different techniques can be found, although all of them utilize
the transformation of starch into sugar followed by fermentation of the resulting sweet wort
(Echeverría and Muñoz, 1988).
The biochemical transformation of cornstarch is presumably one of the most amazing
techniques developed by Andean peoples a long time ago. As previously stated, germination
of the grain is a strategy used to attain sugars from starch in the production of “chicha de
jora”. However, Indians frequently used to chew the corn to produce a soft mass rich in
amylase enzymes from saliva. Figure 1 shows a diagram of the chicha production and the
most common variations. Moreover, in some cases the addition of hallucinogenic flowers,
such as Datura sp., flavoring wild fruits, or herbs, has been reported by chroniclers from the
XVI Century (Burgos, 1977).
Crossbred “chicheros”, living near the town of Riobamba during recent years (1950s to
1970s), used to employ human stools, bones, and other organic wastes, to activate the
fermentation process in order to obtain a stronger chicha. This practice is condemned for its
terrible health and mental consequences to consumers. This pernicious kind of chicha was
used by “chicheros” as a domination tool and a power instrument over the Indians (Burgos,
1977).
Why use Stools for Fermentation?
The use of stools in the fermentation process remains unexplained, but it is probably the
result of a primitive utilitarian practice that became a sort of ceremonial cult practice in
fermentation. Currently, in Cochabamba, Bolivia, the addition of human feces to the chicha
fermenter vessel remains a common practice, with the goal of obtaining a stronger alcohol
intoxication effect (E. Claros, personal communication).
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Fátima C. O. Gomes, Inayara C. A. Lacerda, Diego Libkind, et al.
Candida species present in stools perform alcoholic fermentation, and this is a plausible
reason for its use, as Indians considered stool addition as a useful practice to enhance ethanol
levels in chicha. In our understanding, the need to achieve a superior level of fermentable
wort sugars by the addition of human intestinal microflora may be due to the shortage or lack
of environmental wild Saccharomyces sensu stricto species in the ancient environment of
Quito, as well as a deficient starch conversion during the mashing or chewing of malted or
unmalted corn.
Saccharomyces paradoxus, the natural parent species of the Saccharomyces sensu stricto
group (which includes S. cerevisiae), are found chiefly in oak exudates and other trees
exudates (Kurtzman and Fell, 1998). The distribution of oak trees in South America is
restricted to Colombia. No oak populations have been found in Ecuador (Gentry, 1996).
These ecological aspects of the Saccharomyces sensu stricto species led us to the
hypothesis that these fermenting yeast species could be rare or even nonexistent in the ancient
environment of Quito. If this hypothesis is demonstrated using molecular and microbiological
techniques, a cultural practice could be explained by an ecological fact, considering the
Bioarchaeological approach.
Figure 1. The process of brewing chicha is very similar to the beer production process. It is considered
as the indigenous Andean beer.
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2.1.2. Ancient Yeasts from Ancestral Chicha
As pointed out previously, pondos used for the fermentation of chicha were made of clay
and the pores were obstructed with hydrated starch. This provided microorganisms with the
opportunity to proliferate in the microhabitats offered by the clay material. In this way, starter
cultures could be saved, and continuous batch fermentations were possible without the
addition of new inoculums. Moreover, yeasts had the possibility to survive in a dormant state
for hundreds or even thousands of years within the microhabitats provided by the pores of the
fermentation vessels. In recent investigations carried out at the CLQCA (Colección de
Levaduras Quito Católica), researchers “resuscitated” yeast strains that were immobilized for
centuries within the pores of clay fermentation vessels found in Indian tombs in the La
Florida necropolis (M. C. Molestina, personal communication).
The methods used to recover and reconstruct the cell walls and cell membranes prior to
plating yeast isolates on Petri dishes were developed for isolation from clay fermentation
vessels. Underlying these techniques is the nascent Bioarchaeology that pursues an
understanding of the ancient microflora and its implications for human beings by using
archaeological rests as sources of ancient microorganisms (Figure 2). Using the resuscitation
method, it was possible to retrieve and isolate dozens of different yeasts, bacteria, and molds.
Yeast isolates that were identified and characterized are cited below. Astonishingly,
Saccharomyces species were absent in the tested vessels. The following yeast species were
found in chicha vessels from the Hipia culture (100 to 640 A.C.) in Quito: Clavispora
luisitaniae, Cryptococcus saitoi, Rhodotorula mucilaginosa, Rhodotorula sp. nov., Pichia
guillermondi, Cr. diffluens, Candida parapsilosis, and C. tropicalis.
Figure 2. Sampling of ancient Indian bowls.
2.1.3. Yeast Dispersion Or Domestication?
It is hard to determine whether chicha yeasts were domesticated or were simply dispersed
by insects or other vectors from the environment to the fermenting vessels. To our
understanding, a combination of both events probably occurred. It is well known that
traditional Indian fermenting kettles pass from one generation to the next. These kettles are
appreciated as especific and unique tools for chicha fermentation because of the quality,
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Fátima C. O. Gomes, Inayara C. A. Lacerda, Diego Libkind, et al.
flavor, and ethanol level attained therein. Microflora immobilized within the kettles carry out
successful and robust fermentation (M. C. Molestina, personal communication).
Microbiological and microsatellite DNA studies of chicha yeast strains collected in
ancient fermentation vessels from 600 to 1200 A.C. show that one particular Pichia
guilliermondii strain is consistently present. This finding suggests a possible strain
domestication. Nevertheless, further studies must be performed to determine the role of this
and other species found in archaeological chicha vessels. It is possible that the inoculum of
fermenting yeasts was shared from one fermenting vessel to the next for a long time. Studies
of S. cerevisiae present in isolates from Asia, Europe, and Africa suggest that wild
Saccharomyces ancestors experienced at least two episodes of domestication—in ancient
China and Africa—throughout history (Fay and Benavides, 2005). Moreover, domestication
of industrial wine yeasts has been studied by Querol et al. (2003), who define domestication
of wine yeasts as the adaptation of yeasts strains to the environmental conditions faced during
the fermentation stages of wine. In this manner, starter cultures could be selected and
dispersed among peoples from distant regions in the same way that S. cerevisiae strains were
selected by Chinese, Sumerians, Babylonians, and Egyptians thousands of years ago
(Hornsey, 2003).
A very remarkable issue is the apparent total absence of Saccharomyces species in
ancient chicha. To the best of our understanding, this fact could suggest that non-
Saccharomyces species fermented corn worts in the distant past. This idea could lead us to the
supposition that Indians were domesticating yeasts other than Saccharomyces species. What
is clear is the fact that wine and beer fermentors sampled in regions of Quito belonging to the
period after the Spanish conquest possess S. cerevisiae, as expected.
2.1.4. Chichas in Patagonia
Aboriginal communities in Andean Patagonia (Argentina and Chile) used to prepare
fermented beverages from several raw sources, including cereals and fruits. The chicha made
from corn was probably the most common form of this beverage in South America, and this
type of chicha was also prepared in Andean Patagonia. The most important aboriginal group
inhabiting the temperate forests of Argentina and Chile was the Mapuche (Mösbach, 1992;
Donoso and Lara, 1996), who have been characterized as a typical gatherer community. They
also used wild fruits, such as strawberries (Fragaria chiloensis), maqui(Aristotelia
chilensis),calafate (Berberis spp.), and others, to produce fermented beverages (Pardo,
2004; Pardo and Pizarro, 2005).
An interesting case was the chicha made from the seeds of the Pehuen tree (Araucaria
araucana), from which the Mapuche´s communities prepared the fermented beverage called
Mudai. Mudai preparation shows similarities with that of other chichas derived from corn.
This beverage has been associated mainly with religious ceremonies, although in such cases
the fermentation in generally avoided and the fresh juice obtained from Pehuen seeds, water,
and sugar is consumed immediately prior to the fermentation process (Pardo and Pizarro,
2005).
No works involving the microbial biota present during Mudai fermentation have been
published until now, probably due to the difficulty that imply the sampling process in the
original communities. However, preliminary results were obtained by fermenting the juice
obtained from Pehuen seeds prepared in the traditional way in sterile flasks under laboratory
conditions (Lopes et al., unpublished results). In that work, the fermentation process was
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Traditional Foods and Beverages from South America: …
monitored by measuring the system weight loss due to the liberation of CO2 produced by
yeast metabolism, according to the methodology described in Lopes et al. (2007a) for wine
yeast evaluation. The complete fermentation occurred in 20 days. Yeast sampling in complete
medium was carried out during the process, evidencing maximum population densities of
1.5x108 cfu mL-1. A very poor morphological diversity was observed among the detected
yeast colonies. Molecular identification of these colonies by using the methodology described
by Esteve-Zarzoso et al. (1999), ITS1-5.8S-ITS2 PCR-RFLP and sequence analysis
(Kurtzman and Robnet, 1998), revealed the presence of only two yeast species at the
beginning of fermentation: Hanseniaspora uvarum and Saccharomyces cerevisiae in
percentages of 80% and 20% of the total biomass respectively. In subsequent stages of
fermentation, the yeast biota corresponded exclusively to S. cerevisiae in all analyzed
fermentations. As it was mentioned previously, the apiculate yeast H. uvarum and other
related species of Hanseniaspora has long been associated with fermentations of different
sugar-rich raw materials including grapes (Nisiotou and Nychas, 2007; Barata et al., 2008),
apples (Suárez Valles et al., 2007; Morrisey et al., 2004), oranges (Las Heras et al., 2003),
cocoa beans (Nielsen et al., 2007), etc. Intraspecific analysis of the S. cerevisiae isolates from
Mudai fermentations by means of mtDNA-RFLP (Querol et al., 1992; Lopes et al., 2002)
evidenced a single restriction pattern. This kind of biological homogeneity is frequently
observed in inoculated fermentations in which the yeast starter selected culture dominate the
fermentation process, but it is not expected in natural processes (Querol et al., 1992; Lopes et
al., 2007b). Traditional production of this beverage does not involve the use of commercial
yeasts; however, the environment in which this product is elaborated is in permanent contact
with commercial yeasts used in bread making. MtDNA-RFLP analysis of commercial bakery
yeasts showed the same molecular pattern detected in Mudai fermentations, evidencing a
cross-contamination of yeasts in these traditional fermented products. The use of commercial
yeasts by Mapuche communities had been previously reported; however, this is the first
evidence from an ecological molecular point of view (Lopes et al., unpublished results).
Mudai fermentation samples obtained from different communities are being currently
analyzed in order to find indigenous Saccharomyces isolates.
Another interesting case is the chicha of the Mapuche community that was not prepared
from a cereal, fruit, or root, but was derived from the stromata of a fungus of the genus
Cyttaria (Ascomycota, Leotiomycetes, Cyttariales). Cyttaria fungi are obligate parasites of
several tree species of the genus Nothofagus (figure 3), endemic to Patagonia (Gamundi,
1971). The presence of the fungus produces a typical globose tumor or canker on the tree
trunk and branches (Gamundi, 1971). In the spring, yellowish globose ascostromata (up to
11cm in diameter) appear on these tumors, and these ascostromata are rich in sugars and
polyols (de Lederkremer and Cirelli, 1988). Several Cyttaria species exist in Patagonia and
differ in size, color, texture, and the Nothofagus trees that they parasitize. These fungi are
generally known as Llao-llao in Argentina, while in Chile they are called digueñe, pinatra, or
curacucha. The Mapuche tribe, whose traditional territory is coincident with the Nothofagus
forests in Patagonia, collected and consumed Cyttaria stromata (Pardo and Pizarro, 2005). C.
espinosae, C. hariotii, C. darwinii, and C. berteroi were probably more commonly used than
other fungal species (Schmeda-Hirschmann et al., 1999; Ladio and Lozada, 2000) because of
their higher sugar content and superior palatability. The stromata were considered as fruits of
the host trees and were not only eaten fresh, but were also fermented to make an alcoholic
beverage (Mösbach, 1992). The production of Cyttaria chicha can be considered as exclusive
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Fátima C. O. Gomes, Inayara C. A. Lacerda, Diego Libkind, et al.
to the Mapuche people given the restricted distribution of these fungi. Thus, Mapuche´s had
the distinctive tradition of preparing chicha from a fungus, a practice rarely or most likely
never found in other regions of the world. Unfortunately, little is known about the social
traditions or rituals associated with the preparation and consumption of Cyttaria chicha in the
pre-Colombian period. Today, to the best of our knowledge, the tradition of preparing
Cyttaria chicha has been lost, although the stromata are still consumed fresh, for example in
salads (Tacon et al., 2006; Ladio and Lozada, 2000). Cyttaria chicha could have been
obtained by squeezing fresh stromata and collecting the resulting juice, which fermented
spontaneously, or simply by leaving the entire stromata in cold boiled water for a few days.
Figure 3. Pictures of Cyttaria harioti stromata on Nothofagus dombeyi tumors that were infected via
insects (left). C. hariotii stromata of different maturation stages (right).
Cyttaria stromata undergo a process of maturation that may take 10 to 12 days,
culminating in a spontaneous fermentation process analogous to that observed for grapes,
mainly in those species with higher sugar content. A few studies have reported the presence
of yeasts that naturally associate with Cyttaria stromata and, altogether, reveal the existence
of a complex diversity of species (Ruffini and van Broock, 1997; Brizzio and van Broock,
1998; Ulloa, 2007; Libkind et al., 2007). In C. hariotii stromata, yeasts of all maturation
stages were found, although they varied in number and yeast diversity. A gradual increase in
yeast counts from immature to mature fruiting bodies was observed. Mature stromata yields
were normally greater than 9x104 CFU(colony-forming units)/g (wet weight), while
immature stromata and those in the initial stages of maturation ranged between 900 and 2,000
CFU/g, respectively (Libkind et al., 2007). The proportion of pigmented (red to orange
colonies) yeasts in C. hariotii gradually declined with maturation. In late maturation stages,
only 1 to 5% of the colonies on the isolation plates were pigmented, and most of the yeasts
present were fermentative ascomycetes of the genera Candida and Saccharomyces (Ruffini
and van Broock, 1997; Libkind et al., 2007). Other fermentative ascomycetes associated with
Cyttaria were assigned to the genera Hanseniaspora, Zygosaccharomyces, Pichia,
Kregervanrija, and Torulaspora (Ulloa, 2007; unpublished results), whereas a few
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fermentative basidiomycetes such as Xanthophyllomyces dendrorhous (Libkind et al., 2008)
and Mrakia spp. (unpublished results), were also found. Despite the great diversity of
fermenting yeast species in Cyttaria, the main producersof fermentation, either in the forest or
during chicha preparation, are most likely Saccharomyces species. Ruffini et al. (1998)
carried out spontaneous fermentations of surface-sterilized Cyttaria stromata in the laboratory
and found that Saccharomyces spp. were the major fermenting agents. When mature, Cyttaria
stromata often contain larvae of insects (i.e., Mycetophialidae; Gamundi and de Lederkremer,
1989), which almost certainly act as vectors of many of the yeast species that are later
involved in the fermentation process. Preliminary results indicate that Saccharomyces yeasts,
known as psichrophylic (i.e., S. uvarum and/or S. bayanus), may be involved in spontaneous
Cyttaria fermentation (Ulloa, 2007; unpublished results). This is in agreement with the low
annual mean temperature (ca. 8ºC) of the Patagonian region.
2.2. Masato
Masato is an important beverage in the diet of indigenous populations of the Amazonian
region of Peru. This product is obtained from cassava roots, which are cooked and crushed to
prepare a mass. The fermentation is spontaneous and lasts between 72 and 96 h at room
temperature. In some communities of Peruvian Amazonia, masato is prepared using
traditional recipes (with the saliva of children or old women) for different objectives, such as
religious, political, and commercial ceremonies (Sotero et al., 1996). This type of beverage is
also produced in Moçambique (Africa) and is called masata.
2.3. Beer
Beer is a fermented wort produced from cereals. In this sense, beer is not only what we
currently perceive as a foamy, sparkling, and frequently transparent beverage, but also
includes a wide range of fermented beverages, from chicha to sake, all of which are made
from cereals as the main source of sugar (Berger and Duboë-Laurence, 2005; Hornsey I.,
2003).Throughout history, nomads discovered that fermentation caused to their liquid food
(made of grains, roots, or fruits) to affect their bodies and minds. Anthropologists share the
idea that alcoholic beverages were very important to these human populations, and evidenced
by cereal production destined for the manufacture of beer in very early civilizations. In
Sumer, beer making reached a high grade of sophistication 5000 years ago. This civilization
allotted a large proportion of their cereal crops to beer production. This production was
important not only for ceremonies and cults, but also to pay salaries Their production were
addressed not only to ceremonies and cults, but also to pay salaries.
The art of brewing was dispersed to Babylon, Egypt, and then to Europe, where the
Vikings were conspicuous brewers and warriors. In the Middle Ages, Catholic monks
achieved a number of advancements in brewing technology. As a result, a number of beer
recipes were created, as well as many different kinds of brews that are currently available,
some of which have attained very significant commercial success, chiefly in Germany and
Belgium (Berger and Duboë-Laurence, 2005). Europe was the center for the development of
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Fátima C. O. Gomes, Inayara C. A. Lacerda, Diego Libkind, et al.
the beer and brewing culture and technology, mainly current Belgium, Germany, the Czech
Republic, Denmark, and the United Kingdom, where most of the technical developments and
recipe diversification took place (Berger and Duboë-Laurence, 2005)
The ingredients necessary to make beer are:
1. A source of starch, amino acids, and peptides, such as barley, wheat, rice, oat, corn,
or other cereals, for conversion into fermenting and body enhancing sugars. Most of
the kernels should be malted (germinated, dried, and toasted) in order to produce the
enzymes necessary to break down cell walls (β-glucans), proteins, and chiefly starch.
It is well known that modern beers are made not only from malted cereals, but also
from unmalted grains, flours, and even sugar from cane. These raw materials known
as adjuncts are used to replace a fraction of the malt, providing “drinkability” and
lowering the production costs of industrial beer.
2. A flavoring, preserving, and aromatic ingredient such as hops, which has been used
for its stimulating and preserving capabilities as well as for the bitter and aromatic
characteristic it provides to the brewing wort. The presence of the so-called alpha
acids and essential oils are responsible for the flavor and aroma. Hops were
introduced to beer makers in the 13th Century (Berger and Duboë-Laurence, 2005).
3. Water is the most abundant ingredient in beer. To produce one liter of beer, it is
necessary to use about 8 to 10 liters of water; not only as an ingredient, but also for
different operations, such as cleaning, cooling, and heating. The quality of water lies
in its chemical composition, namely, the presence of diluted salts. In this sense, water
could be classified as hard and soft, each of which is used for different kinds of
beers.
4. Yeast is one of the most important ingredients in beer making, as this microorganism
is not only responsible for the production of alcohol and carbon dioxide, but also a
number of compounds such as esters, aldehydes, and alcohols that give different
kinds of beers their special characteristics. The most important yeast species for beer
making is S. cerevisiae, with a wide variety of strains adapted to the fermentation
process in a no less wide variety of beers. Other species can produce fermentation in
some artisanal beers, such as Dekkera bruxellensis, Dekkera lambicus, and
Schyzosaccharomyces pombe. These non-traditional brewing yeasts ferment special
brews in Europe and Africa (Briggs et al., 2004).
2.3.1. Manufacturing
Brewing production is based on technology dealing with chemical and biochemical
transformations. To produce beer it is necessary to attain a mixture of grains (frequently malt
with other unmalted cereals) and water in proportions that vary from 2 L/kg of grain to 5
L/kg. Mashing takes place in a kettle in which enzymatic transformation converts the
macromolecules (starch and protein) into shorter chains or simple monomers. The mashing
temperature frequently varies from 45 to 70 °C to allow the proteases and amylases to act on
their corresponding substrates. The mashing process takes place for approximately two to
three hours.
Once the saccharification is complete, a filtration step is performed so that clean wort is
obtained. Husks and other solids are recycled as animal feed and, more recently, are used as a
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Traditional Foods and Beverages from South America: …
feasible lignocellulosic material with a wide range of biotechnological applications (Briggs et
al., 2004).
Next, the wort is boiled. During this part of the process, three objectives are achieved:
first, the wort acquires chemical stability while high molecular weight proteins precipitate;
second, hops are added, triggering the heat-mediated extraction of compounds to confer
flavor and aroma; finally, the wort is concentrated and caramelized to adjust the sugar content
and color.
Fermentation and maturation represent the longest period of beer production. During this
step, sanitized kettles contain the wort that was previously inoculated with pure yeast.
Fermentation is divided into two phases: principal fermentation, when most of the
fermentable sugars are transformed into ethanol and carbon dioxide, and secondary
fermentation tightly linked with maturation, which can take as much as the 60% of the beer
production time. An intricate conjunct of the biochemical reactions transforms the wort, a
sweet-bitter liquid, into a foamy, tasty beer.
2.3.2. The First Beer In America
Quito was founded in 1534 by the Spanish conquerors. One of the first constructions built
by the Europeans was the San Francisco Convent (Figure 4), regented by the Franciscan
Order (S. Moreno, personal communication). Even though most of the monks were from
Spain, one was born in Malines, Belgium. Fray Jodoco Ricke was not only a religious man,
but he was also a brewer. Thus, he introduced wheat and barley to America. At the same time,
he imported a brewery directly from Europe whose fermentation vessels were made of oak
wood and whose brew house was made of copper. In 1566, the first brewery in America
initiated the production of top fermenting beer, which represented all beers produced at that
time around the world. Franciscans were probably using European brewing yeast strains,
which could be transported in vessels containing starter beer.
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Fátima C. O. Gomes, Inayara C. A. Lacerda, Diego Libkind, et al.
Figure 4. Mash tun in San Francisco Old Brewery, Ecuador.
Although yeasts were not considered as microorganisms, the ancient brewers knew that
the properties generated by the cream as a “by product” of fermentation was fundamental to
trigger the transformation of sweet wort into beer. Therefore, it is very plausible that the
correct brewing yeast was carried from Europe to America. The San Francisco Convent
brewery terminated operations in the 1970s, after more than 400 years of continuous
production in the same fermenting vessels. It has since been restored and the equipment is
shown to special guests (Papazian, 1992). As part of a survey on the history of fermentation
in Quito, researchers of the CLQCA were able to resuscitate yeasts from the old brewery.
Samples were taken from the wooden barrels, at different depths from the surface of the
wood, by scratching the insides of the barrels to recover immobilized dormant yeasts.
The Saccharomyces cerevisiae isolates recovered from these barrels are of special
interest for their biological, historical, and cultural importance. Further studies on mtDNA
restriction profiles will provide novel data about the yeasts used in the past by monks to make
the oldest beer in America. Similar discoveries could be carried out for fermentors worldwide
to provide new data to researchers that study human history via yeast dispersion and
domestication (Mc Govern, 2004; Fay and Benavides, 2005).
2.4. Cachaça
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Traditional Foods and Beverages from South America: …
Cachaça is the most traditional alcoholic beverage in Brazil, and it is made by the
distillation of fermented fresh sugarcane juice. It was first made from the waste of sugar
extraction from sugar cane, when Brazil began producing sugar in 1534. The first raw
material used to make cachaça was the foam formed during the concentration of cane juice
removed from the heating kettles. After spontaneous fermentation, this material was first
distilled in small alembics (pot stills) made from clay by the Portuguese between 1534 and
1550 using their traditional knowledge of aguardente or bagaceira production from grape
pomace. The production of cachaça was first intended for the slaves, but the growing
approval of the new beverage stimulated producers to make more and to start fermenting the
cane juice itself. By the beginning of the 17th century, cachaça and sugar were already
commercialized all over the northwest Brazilian coast, and they were bartered for slaves by
individuals on the Brazilian ships traveling to Africa (Carvalho and Silva, 1988).
Until the end of the Second World War, cachaça continued to be a product made by a
large number of small producers who planted sugar cane, produced cachaça, and traded their
own products. In the postwar period, this small-scale production began to face competition
from large, continuous distillation plants and new bottling companies with their own
commercial brands (Lima, 1983). Now, with an estimated production of about 1.3 billion
liters per year, Brazilian cachaça is the third highest consumed distilled liquor in the world.
Therefore, its consumption, which was initially essentially local, is now influenced by larger
factories as this beverage reaches more and more new markets around the world (Faria et al.,
2003).
The old regulations concerning sugar cane spirits were finally consolidated by
government decree 2314 (Brasil, 1997), which defines the spirits made from fermented sugar
cane juice by the terms cachaça, caninha, or aguardente de cana – and by decree 4072, which
reserved the term cachaça only for the sugar cane spirit made in the Brazilian territory (Brasil,
2002). According to Brazilian legislation, cachaça is a distilled alcoholic beverage comprising
38 to 48% alcohol (v/v) at 20oC, produced by distilling fermented sugar cane juice, to which
no more than 6 grams of sugar may be added per liter for taste correction. When the amount
of sugar added is 6 to 30 grams per liter, the beverage is called ‘sweetened cachaça’.
In Brazil, Minas Gerais state is the main center of production of traditional cachaça
(“alembic cachaça”); however, the beverage is produced in all regions of Brazilian. Most of
the distilleries produce cachaça by spontaneous fermentation, conducted by the action of the
microbiota found in the sugar cane juice (Figures 5 and 6). These microorganisms can also be
associated with the equipment (mill and vats) or can be introduced by insects that visit the
production area (Morais et al., 1997). In addition to spontaneous fermentation, some
producers use bakery yeasts or dry yeasts from the industrial ethanol production as starters of
the fermentative process (Rosa et al., 2009). The initial process in the traditional natural
starter ferment is conducted in a vat and can last 5 to 30 days. Some additional materials can
add to the sugar cane juice, such as powdered corn and rice, citric fruits, soy, and others.
When the natural ferment is ready to use, fresh sugar cane juice diluted to 14 to 26oBrix is
added to the vat to initiate the fermentative cycles. Each fermentative cycle lasts between 20
and 24 hours. This batch fermentation is carried out at room temperature, which, depending
of the Brazilian region, can vary between 15 and 40oC. At the end of each fermentative cycle,
the sugar cane wine is distilled in copper alembics and fresh sugar cane juice is added to start
a new fermentation. The same starter ferment can be used for the entire production season
(between May to November). Commercial yeast grown for ethanol production or baking are
15
Fátima C. O. Gomes, Inayara C. A. Lacerda, Diego Libkind, et al.
utilized for cachaça fermentation in larger industrial distilleries. Industrial distilleries utilize
centrifugation to carry out cell removal and recycling, although some do not resort to this
process, and the yeasts take an average of four hours to spontaneously flocculate to the
bottom of the vats. Traditional fermentation is conducted in smaller distilleries that have a
maximum production capacity of 1,000 liters of cachaça per day. These traditional distilleries
utilize a natural starter culture prepared by spontaneous fermentation, and they do not use
centrifugation for cell recycling.
The sugar cane juice is fermented by a complex microbial community. For each addition
of fresh sugar cane juice and dilution water, new populations of yeast and bacteria are
introduced into the fermentation environment. These microorganisms multiply and modify
the must characteristics, influencing the frequency of occurrence of the microbial populations
during the fermentation. Saccharomyces cerevisiae is the most prevalent yeast species present
during the fermentative cycle, but minor populations of non-Saccharomyces yeasts are
frequently found (Morais et al., 1997; Pataro et al., 2000; Guerra et al., 2001; Schwan et al.,
2001; Gomes et al., 2007; Marini et al., 2009). During fermentation, the yeast cells are
subjected to a great number of adverse conditions, such as osmotic stress due to the high
concentration of sugars in the sugar cane juice; low or high environmental temperatures; and
the high concentration of alcohol (8% v/v) at the end of the fermentative cycle. These
different stresses affect the survival of the yeast strains during fermentation, and temperatures
above 35oC and a high alcohol concentration are indicated as stress factors that influence
yeast fermentation. These stressful conditions have a strong selective pressure on the
microbiota responsible for the fermentation (Guerra et al., 2001).
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Traditional Foods and Beverages from South America: …
Figure 5. Sugar can mill (A), cleaning tanks of sugar cane juice (b), Preparation of the starter natural
ferment and fermentations vats for traditional Cachaça production.
Lactic acid bacteria are regarded as contaminants of cachaça fermentation. They can also
deteriorate harvested sugar cane, thereby reducing the sugar content (Gallo, 1992). These
contaminant bacteria compete with yeast for sugar substrates and micronutrients and produce
end products, such as lactic and acetic acids, that inhibit yeast growth and metabolism, which
reduces the ethanol yield of fermentation. Contaminants can arise from tankage, heat
exchange, raw materials, active dry yeast or other yeast preparations, poorly stored backset,
recycled, or continuously propagated yeast slurries used as inocula, inadequate cleaning-in-
place procedures, and inadequate yeast-conditioning (propagation) times and/or temperatures
(Narendranath and Brey, 2009). The occurrence of 1 X 107 lactobacilli per mL of the initial
mash results in an approximately 1% v/v reduction in the final amount of ethanol produced by
the yeast, depending on the strain of contaminant bacteria (Narendranath et al., 1997). Lactic
acid bacteria are able to grow rapidly and survive under ethanol production conditions. While
most bacterial species do not survive in the presence of over 5% v/v ethanol in the medium,
some species of lactobacilli are able to grow even in the presence of more than 15% v/v
ethanol present in substrates (Narendranath and Brey, 2009).
Figure 6. Copper alembic (A), fermentation must (B), fermentation vats (C), and tuns (D) for the
storage of Cachaças.
The use of selected S. cerevisiae strains constitutes an alternative way to more efficiently
control the cachaça fermentation process (Rosa et al., 2009). These yeast strains could
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Fátima C. O. Gomes, Inayara C. A. Lacerda, Diego Libkind, et al.
improve the sensorial attributes of traditional cachaças, which are very important for the
expansion to new markets of this beverage (Gomes et al., 2007; Bernardi et al., 2008).
2.5. Champús
Champus is a popular cereal-based low alcoholic beverage with a sweet and sour taste
that is widely consumed in rural and urban areas of Colombia and in some other South
American countries, including Ecuador and Peru (Osorio-Cadavid et al., 2008). The
production of champu´s utilizes different cereals, such as wheat, rye, and maize or their
combinations together with other ingredients such as pineapple or ‘‘lulo’’ (Solanum
quitoense Lam.), ‘‘panela’’ (sugar cane) syrup, clove, cinnamon, and/or orange tree leaves.
The popularity of this drink is due to its pleasant taste and flavor, as well as its nutritional
properties (Osorio-Cadavid et al., 2008).
According to Osorio-Cadavid et al. (2008), Colombian Champu´s is produced by boiling
corn kernels for many hours in order to soften them; then, after cooling the grains at room
temperature, fruits are added together with ‘‘panela’’ syrup (4–6%) and the other ingredients.
The beverage is stored at about 12–15oC and consumed within 24–48 h. During this time,
fermentation occurs and a beverage with a low alcoholic content (2.5–4.2% alcohol content
and pH values between 3.5 and 4.0) is obtained, depending on the quantities of fruit added.
Yeast species associated with Colombian Champu´s fermentation were studied by
Osorio-Cadavid et al. (2008), and include S. cerevisiae, Issatchenkia orientalis, Pichia
fermentans, P. kluyveri var. kluyveri, Zygosaccharomyces fermentati, Torulospora
delbrueckii, Galactomyces geotrichum, and Hanseniaspora spp. Selected strains of these
yeasts could provide a useful contribution to the improvement of flavor and to product
acceptability. However, the role of lactic acid bacteria associated with this traditional
fermentation needs to be determined.
2.6. Cauim
Many of the fermented foods used by Brazilian Indians, however, are unknown outside of
the region in which they are produced. The Tapirap´e people of the Tapi’ita˜wa tribe make
nonalcoholic beverages, which are fermented from cassava and other substrates using a
variety of techniques and processes, resulting in a beverage namedcauim’ (Melatti, 1983;
Almeida et al., 2007). Several substrates are used by the Tapirap´e to produce ‘cauim’,
including rice, cassava, corn, peanuts, cottonseed, ‘bacaba’, murity palm, banana, pumpkin,
seeds of wild banana, and other suitable fruits. The production of these fermented products is
based on old empirical knowledge, which is transferred from generation to generation
(Melatti, 1983; Almeida et al., 2007). The Tapirapé Indians make nonalcoholic beverages, for
which the fermentation process is initiated from ingredients other than cassava substrates
using different techniques and processes, resulting in a beverage named ‘cauim’ (kawí)
(Baldus, 1970; Almeida et al., 2007). To prepare ‘cauim’, beverage cassava roots are allowed
to ferment for three to five days in running water to soften the skin. The cassava tubers are
peeled, cut in small pieces, and sun dried. The dried pieces are then grated into flour. This
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Traditional Foods and Beverages from South America: …
flour is mixed with water and cooked for about 2 h and then cooled at room temperature.
When the porridge is cold, an inoculum is added to initiate the fermentative process, which
usually takes 24 to 48 h. The inoculum is obtained from a fluid resulting from chewing the
sweet potato (performed by female Indians) and the saliva of women who have been chewing
sweet potato is used to prepare ‘cauim’ (Almeida et al., 2007). The saliva inoculum is
generally used to initiate fermentation regardless of the substrate used. The saliva–sweet
potato mixture is added slowly to the substrate for beverage production, and this permits the
growth and multiplication of microorganisms (Schwan, 2004; Almeida et al., 2007). Almeida
et al. (2007) reported that during the fermentation of cauim, lactic acid bacteria increase for
the duration of the process to become the dominant microorganisms in the ferment.
Lactobacillus pentosus, L. plantarum, Corynebacterium xerosis, C. amylocolatum, C.
vitarumen, Bacillus cereus, B. licheniformis, B. circulans, and Paenibacillus macerans were
found. The species L. pentosus and L. plantarum were the dominant bacteria and were present
during all periods of fermentation. According to these authors, no hygiene control or proper
technology are associated with the cauim produced by Tapirapé Amerindians, and the
identification of the microbiota associated with this fermentation may be used to improve the
quality of the final product as well as to define a starter culture to replace the traditional
inoculum.
2.7. Wine
Wine is the product obtained from the alcoholic fermentation of fresh grape juice.
According to historians, the first reports about wine in human history refer to the Caucasus
and Mesopotamia in 6000 BC. Wine was later mentioned in Egypt, Phoenicia, Greece, and
Crete. Romans spread winemaking all around the Mediterranean, where it has been produced
since 500 BC. In the sixteenth century, European conquerors introduced the vine into the New
World, particularly in the recently colonized territory of Mexico (Pretorius, 2000). Because of
the success of this new culture, vineyards were also introduced into Peru and more recently
into other South American countries, particularly Chile and Argentina. Today, the most
important wine regions in South America are the central areas of these two countries.
As previously mentioned in this chapter, the yeast genera Saccharomyces have been
related to all alcoholic fermentation processes, and wine fermentation is no exception. The
yeast species Saccharomyces cerevisiae in particular and, to a lesser extent, Saccharomyces
uvarum have long been considered the key microorganisms in this process. Therefore, the
most modern, industrialized winemaking processes use selected cultures (starter cultures) of
these yeast species (particularly S. cerevisiae) to drive the process. Consequently, wines
obtained using selected grapes and yeasts always exhibit the same reproducible physical-
chemical and aromatic characteristics. Today, more than 200 S. cerevisiae starter cultures are
being produced as active dry yeasts (ADY) for the commercial wine market.
Novel ecological research has revealed that the microbiology of wine production extends
beyond the activity of this single species and involves the contribution of a great variety of
microorganisms, including numerous yeast species, filamentous fungi, lactic and acetic acid
bacteria, and others (Fleet, 2003). Moreover, the use of modern molecular methods for yeast
strain characterization, such as restriction fragment length polymorphism of the mitochondrial
DNA (mtDNA-RFLP), pulsed-field gel electrophoresis (PFGE), random amplification of
polymorphic DNA (RAPD), PCR amplification of delta elements, and microsatellite typing,
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Fátima C. O. Gomes, Inayara C. A. Lacerda, Diego Libkind, et al.
have been applied to wine strain characterization and revealed an extraordinarily high genetic
variability between wild isolates of S. cerevisiae present in wine fermentations (Querol et al.,
1992; Vezinhet et al., 1992; Baleiras Couto et al., 1996; de Barros Lopes et al., 1996;
Hennequin et al., 2001; Pérez et al., 2001; Lopes et al., 2002). Based on the detected
ecological complexity of wine fermentation processes, several yeast mixed cultures have been
proposed as starter cultures (Fleet, 2008); however, the use of these products are still
restricted, mainly because of the difficulty associated with ADY production for non-
Saccharomyces yeasts.
The basic steps in winemaking can be summarized as: (i) grape crushing and extraction
of the juice, (ii) alcoholic fermentation by yeasts, (iii) optional malolactic fermentation by
lactic acid bacteria (this fermentation can take place simultaneously or after alcoholic
fermentation), (iv) storage and aging, and (v) packaging and selling. As previously mentioned
for alcoholic fermentation, selected cultures of lactic acid bacteria (Oenococcus oeni) are also
available on the market for conducted malolactic fermentations.
Both spontaneous—based on the fermentation of grape juice via the indigenous yeast
microbiota naturally present on grapes or winery surfaces—and conducted winemaking
processes—in which selected starter yeast cultures are inoculated into the must—are evident
in different wineries in South America. Inoculated wine fermentations are the preferential
choice for most industries because of the reproducibility and homogeneity of the final
product. As most yeast starters have been isolated and selected in old world winemaking
areas, the yeast diversity found in these types of fermentations is not representative of the real
diversity that might be detected in spontaneous or natural processes in South American
countries (Lopes et al., 2007b; Mercado et al., 2007). Nevertheless, the most interesting
feature of wine fermentations in South America is the fact that spontaneous fermentations are
still carried out by several wineries in different winemaking areas, particularly as the selected
methodology in small familial cellars. This fact, as well as the geographic distance from the
big wine centers of the world where inoculated fermentations are the habitual methodology
for wine fermentations, makes South America an interesting habitat for studying the diversity
of yeasts associated with this complex ecological process (Martínez et al., 2007).
In contrast to the scarce information concerning microbial yeast diversity in the
previously discussed traditional fermentations, a large number of scientific publications have
focused on the biodiversity associated with grape surfaces, grape must, and winery surfaces in
several winemaking regions of the world, including specific areas in South America.
However, the use of different methodological approaches makes comparison of the results
difficult. These different methodologies comprise the use of complete isolation media
(Pramateftaki et al., 2000; Povhe-Jemec et al., 2001; Sabate et al., 2002; Ganga et al., 2004;
Combina et al., 2005b; Raspor et al., 2002 and 2006; Sturm et al., 2006), selective isolation
media (Hierro et al., 2006; Nisiotou and Nychas, 2007; González et al., 2007; Lopandic et al.,
2008; Barata et al. 2008), or even culture-independent techniques (Prakitchaiwattana et al.,
2004; Di Maro et al., 2007; Renouf et al., 2007). Moreover, different methodological
approaches, including the use of different grape varieties, different temperatures and SO2
levels, vineyards employing different technological treatments, and traditional or molecular
(different types) identification methods, have also been used. It is generally agreed that the
populations of microorganisms on the surfaces of mature grapes is about 103–105 CFU/g,
consisting mostly of yeast and various species of lactic and acetic acid bacteria (Fleet, 1999).
The yeast species H. uvarum, C. stellata/C. zemplinina, and M. pulcherrima and the yeast-
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Traditional Foods and Beverages from South America: …
like Aureobasidium pullulans are particularly abundant on grape surfaces worldwide
(Prakitchaiwattana et al., 2004; Sturm et al., 2006; Raspor et al., 2006; Nisiotou and Nychas,
2007; Renouf et al., 2007). To a lesser extent, yeast species belonging to the genera Pichia
(P. guilliermondii, P. membranifaciens), Cryptococcus (Cr. ater, Cr. laurentii, Cr. albidus,
Cr. magnus), Rhodotorula (R. glutinis, R. mucilaginosa), Candida, Issatchenkia,
Kluyveromyces, Torulaspora, and Zygosaccharomyces are also normal inhabitants of these
surfaces. Populations of the strictly aerobic yeast species, such as those belonging to the
genera Cryptococcus and Rhodotorula as well as to A. pullulans, rapidly decline after grape
crushing. In contrast, fermentative yeast species like H. uvarum, C. stellata/C. zemplinina,
and M. pulcherrima become the most abundant yeast species during the initial stages of
alcoholic fermentation (Pramateftaki et al., 2000; Povhe-Jemec et al., 2001; Raspor et al.,
2002; Sabate et al., 2002; Hierro et al., 2006; González et al., 2007; Lopandic et al., 2008).
These yeast species rapidly consume a high quantity of sugars (more than 200 g/L of a
proportional mix of glucose and fructose), producing a moderate amount of ethanol. Under
these highly selective conditions, S. cerevisiae becomes the predominant species until the end
of fermentation, even when its numbers on the grape surfaces are extremely low. In South
America, the diversity of yeasts and bacteria associated with grapes, wine fermentations, and
winery surfaces have only been described in the last two decades. Nevertheless, the yeast
diversity present on grapes (Combina et al., 2005a) and in spontaneous wine fermentations in
South America (Ganga et al., 2004; Combina et al., 2005b) seems to be similar to the
diversity observed in other winemaking regions of the world.
In inoculated fermentations, the diversity and abundance of non-Saccharomyces wine
species are drastically reduced, as observed in different wineries in Chile (Ganga et al., 2004),
Argentina (Lopes et al., 2007b) and the old world (Xufre et al., 2006; Andorra et al., 2008;
Zott et al., 2008).
Wine yeast diversity studies and selection programs have been carried out in recent years
in different winegrowing areas of South America, including the central area of Chile (Ganga
et al., 2004; Martínez et al., 2004 and 2007), the Argentinean areas of Mendoza (Combina et
al., 2005a and b; Mercado et al., 2007) and North Patagonia (Sangorrín et al., 2001; Lopes et
al., 2002; Rodríguez et al., 2004; Lopes et al., 2007a; Lopes et al., 2007b; Sangorrín et al.,
2007), and the less documented regions of San Juan in Argentina (Toro and Vazquez, 2002),
Uruguay (González Techera et al., 2001), and South Brazil (Silva, 1996; Silva et al., 2006).
All of these studies are very important to protect that indigenous yeast biota, as well as to
develop regional yeast starters that are better adapted to the local ecological characteristics of
these particular winegrowing regions. Previous studies carried out in South America have
evidenced that commercial starter cultures introduced into local vinification processes are
also detected in spontaneous fermentations carried out in the same winery or on the winery
surfaces (Mercado et al., 2007; Martínez et al., 2004 and 2007). These starter cultures reached
even the vineyards and could also be detected in future fermentations (Schuller et al., 2005).
3. FERMENTED FOODS PRODUCED IN SOUTH AMERICA
3.1. Traditional Minas Cheese
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Fátima C. O. Gomes, Inayara C. A. Lacerda, Diego Libkind, et al.
There are several kinds of traditional cheeses produced in South America, such as: Minas
cheese (discussed below): Coalho cheese, made with semi-cooked curd produced in the
northeast of Brazil; Serrano cheese, produced with crude milk in the south of Brazil; Pategrás
cheese, a partially cooked cheese, which optionally can contain small eyes that are well
distributed throughout the cheese, that undergoes a ripening period ranging from 30 to
60 days (Perotti et al. 2009); Corrientes artisanal cheese, which is manufactured by local
farmers on a small scale for decades using raw milk from Argentine Criolla cows and
traditional techniques passed down from generation to generation, is commonly consumed by
most of the northeastern population of Argentina (Vasek, 2008); Reggianito Argentino
cheese, which is made from milk produced by pasture-fed cows. We will discuss the
technological process used for the production of Brazilian Minas cheese as an example of a
very popular traditional cheese consumed in South America.
Minas cheese is a dairy product that is very popular in Brazil (it is the third largest cheese
manufactured, after mussarela and “prato” cheese). Minas Gerais state is the most important
producer of traditional Brazilian cheeses (Fonseca et al., 1995). This is a semi-hard cheese
produced with pasteurized or unpasteurized cow milk and has a pale cream color, yellowish
rind, cylindrical shape, and homogeneous texture (Borelli et al., 2006). There are several
varieties of Minas cheese, and the most famous are the Canastra and Serro cheeses. These
cheeses are produced with raw cow’s milk and have been manufactured in a traditional
empirical manner for more than 200 years. The technology employed for the production of
these cheeses was introduced by Portuguese immigrants in the late eighteenth century,
following the techniques used for the manufacture of Serra da Estrela cheese (Silva and
Castro, 1995; Borelli et al., 2006). The production process involves the use of raw cow’s milk
and natural whey cultures as starters and commercial rennet. The natural starter comprises
indigenous lactic acid bacteria associated with the milk and equipment. This natural starter is
termed “pingo” and is represented by the natural whey that drops from cheese covered with
salt from previous procedures. Cheese making takes place in a room adjacent to the cow barn
where the raw milk is filtered from the milking bucket into the cheese vat (Borelli et al.,
2006). The first step in the manufacture of these cheeses is the addition of calf rennet and a
natural starter culture into the milk. The native microbiota produce acid, and after about 1h
the coagulum is cut and transferred to plastic moulds where the whey is removed by hand
pressing. Salt is then added to cover the cheeses for a period of about 6 to 8 h, and the cheeses
are inverted and salted again for another18 to 20 h. After this period, the salt is removed and
the cheeses are ripened in wooden shelves without temperature and humidity controls. The
ripening period lasts between 3 to 15 days (Borelli et al., 2006; Lima et al., 2008).
The implementation of “Good Manufacturing Practices” in the production of Minas
cheeses has been fundamental for preventing contamination. The use of raw milk with a long
ripening period is generally not recommended because the Brazilian climate and the
conditions involved in milk production are favorable for contamination and the development
of microorganisms (Pauciulli, 1996; Lima et al., 2008). Brazilian legislation (Brasil, 1998)
prohibits the production of cheese from raw milk, except if the ripening time is greater than
60 days. However, this recommendation is not followed by the population in general, and
these cheeses are one of the most appreciated dairy products in Brazil.
Lactic acid bacteria and yeasts are the major components of the natural microbiota
associated with the production of Minas cheese. Species of Lactobacillus, Lactococcus, and
22
Traditional Foods and Beverages from South America: …
Streptococcus, with counts of approximately 8 log c.f.u. ml-1, are present in the natural starter
(“pingo”). Lactobacillus plantarum and Lactococcus lactis are the prevalent species
associated with the natural starter and with cheese ripening. The yeast counts are between 1.7
to 7.9 log c.f.u g-1 in the cheese samples, and the most prevalent species are Debaryomyces
hansenii, Kluyveromcyes lactis, Torulaspora delbrueckii, Kodamaea ohmeri, and
Zygosaccharomyces rouxii (Borelli et al., 2006; Lima et al., 2009). These species, with the
exception of K. ohmeri, have been isolated from different cheeses produced in other countries
(Borelli et al., 2006). However, the role of these microbial species in the manufacture of the
traditional Minas cheese needs to be determined.
3.2. Cassava
Cassava, Manihot esculenta, Crantz is an enlarged root that is indigenous to the Northern
Amazon, South America and is widely cultivated in tropical countries. Cassava tubers are
mainly used for human nutrition as fermented products, and therefore are a staple food and
the main subsistence crop in the tropics. Cassava is an important substrate for many
fermented foods that are widely consumed in Brazil, Asia, and Africa (Ray and Sivakamur,
2009). The cassava roots contain high levels of starch (85%) and minor amounts of proteins
(1.5%), lipids (0.3%), and other nutrients (Schwan, 2004).
The main characteristic of the cassava root is the presence of complex cyanogenic
glucosids (linamarine and lotaustraline) that, in specific situations, can produce free cyanide,
which, in the presence of water, forms cyanidric acid (Cereda, 2005). All parts of the plant,
including the roots, and mainly the leaves, contain these potentially toxic compounds. Several
food products are produced from the cassava, such as cassava floor, sour cassava starch, and
“tucupi”. The world production of cassava from roots in 2004 was approximately 202.7
million tons.
Cassava fermentation processes are adequate for the reduction of its toxic cyanogenic
glucosides for preservation and for improving the flavor and aroma of the product (Holzapfel,
1997). In Brazil, the indigenous knowledge of ethnic persons living in the northern and
eastern regions on the production of fermented food is worthy of documentation. Several
Brazilian tribes (Araweté, Kayapó, Karajá, Javaé and Tapirapé) utilize small-scale
fermentation for root preservation and for the elaboration of cassava fermented dishes and
foods. Some tribes use fermentative processes to produce food and beverages with nutritional
value, as a stimulant, and for medical and mythical purposes (Wagley, 1988). Foods produced
by Brazilian Indians using cassava, such as cassava flour starch (Carvalho et al., 1999;
Lacerda et al., 2005), ‘puba’ flour (de Almeida et al., 1993), and ‘beiju’, contribute
significantly to the diet of the Brazilian population.
Among the artisanal fermented products of Indian Brazilian heritage ispuba flour,
which is obtained after a fermentative process called pubagem”. This process consists of the
immersion of root cassavas in water [in tanks or traditionally in igarapés (branches of small
Amazonian rivers)] with or without peel and their incubation at environmental conditions.
This period can vary from three to seven days, depending on the cassava characteristics,
composition, concentration of the initial microbiota, temperature and water pH, age of the
roots, and intensity of fermentation (Folegatti et al., 2005). During fermentation, in addition
the degradation of cynogenic compounds and the production of aromatic substances,
23
Fátima C. O. Gomes, Inayara C. A. Lacerda, Diego Libkind, et al.
softening of the roots often occur. Bacteria, yeasts, and molds were identified in the
fermentation ofpubá” and found to be responsible for the production of the acetic, butyric,
and lactic acids that are prevalent during the maceration phase of the cassava root. The
maceration begins 48 h after initiation of the fermentation process. During maceration, the
cassava peel is eliminated and the root loses its consistency, allowing its manual crushing in
wood tanks. The fermentation water containing toxic substances is discarded, and the swollen
roots are sieved to remove non-fermented pieces of roots. The cassava dough obtained
following fermentation is washed with water and again sieved to obtain a product that can be
consumed moist, or sun or oven-dried (Almeida, 1992). During fermentation, the microbiota
initially comprises enterobacteria and corynobacteria, which are slowly substituted by lactic
acid bacteria, sporulated bacteria, and fungi (Candida, Saccharomyces, Aspergillus, and
Penicillium species). The end of fermentation is an empiric process based on the experience
of the producer, who determines visually or by tact the degree of root swelling.
Other traditional cassava fermentation techniques widely used in Latin America,
especially in Colombia and Brazil, provide products obtained by the fermentation of moist
starch extracted from the roo. This product is known as “almidon agrio” in Colombia and
“polvilho azedo” in Brazil (Dufuor et al., 1995; Lacerda et al., 2005). The cassava starch is
extracted by washing, peeling, and grating the roots, and then by placing the paste in
abundant water to release the starch granules and separating them from the fibers and soluble
components (Lacerda et al., 2005). This cassava starch is fermented in tanks for between 20
and 70 days. The process is a semi-solid fermentation, with the formation of compact starch
blocks due to water evaporation during fermentation. The fermented starch is sun dried
following the fermentation process. The starch flour is used for the production of fried goods,
traditional cheese breads in Brazil, and other baked goods.
Fermentation for sour cassava starch production occurs via the action of lactic acid
bacteria, the prevalent microorganisms associated with this process (Figueroa et al., 1995;
Lacerda et al., 2005). The beginning of fermentation is characterized by a drastic drop in the
concentration of dissolved oxygen provoked by amylolitic aerobic bacteria that are able to
consume oxygen and produce gases, such as carbon dioxide and hydrogen (Cereda and
Bonassi, 1985; Camargo et al., 1988). The low availability of oxygen and the products
resulting from the starch hydrolase stimulate acidogenic microbial fermentative pathways,
causing an increase in acid production (Figueiroa and Chuzel, 1991) and oscillations in the
sugar concentrations (Cereda and Lima, 1985). Lacerda et al. (2005) showed that
Lactobacillus plantarum and Lactobacillus fermentum were the prevalent lactic acid bacteria,
at numbers between 6.0 and 9.0 log cfu g-1, during starch fermentation in two factories in
Brazil. Lactobacillus perolans and L. brevis represented minor fractions of the population.
Galactomyces geotrichum, Issatchenkia sp., and Candida ethanolica were the prevalent
yeasts, at 5.0 log cfu g-1.The native microbiota is responsible for the production of the
characteristic flavor of the sour cassava starch.
3.3. Cocoa
Probably originating in Mesoamerica, chocolate or cacao had previously been used as a
food, beverage, and as medicine for over 2,000 years (Schwan and Wheals, 2004). Cocoa is a
very important ingredient in a number of foods, such as cakes, biscuits, child-foods, ice-
24
Traditional Foods and Beverages from South America: …
creams, and sweets. Cocoa beans, originating as seeds in the fruit pods of the tree Theobroma
cacao, are the source of cocoa powder and come from Africa and Central and South America
(Sánchez-Hervás et al., 2008). The principal varieties are Criollo, now rarely grown due to its
disease susceptibility, Forastero from the Amazonas region, and a hybrid, Trinitario, with the
latter two forming most of the “bulk” market. The Arriba type, possessing a “fine” flavor, is
grown in Ecuador (Schwan and Wheals, 2004). The annual world production of cacao is
approximately 2.5M tons.
The ancient Mayan language, century XVI, had two words, Kab (juice) and kaj (bitter),
which together form the Word kabkaj. The beverage prepared with this juice of Kabkaj was
called kabkajatl, since the suffix atl means liquid or water. The Spanish conquerors, due to
pronunciation problems, added the syllable hu from the Indian word, and the word
transformed into kabkajuatl. Later, the orthography of South America people modified the
name to cacauatl (Oetterer et al., 2006). Cacauatl was a beverage prepared without sugar and
consumed cold. The Spanish introduced milk and sugar and began to consume it warm. From
this originated the word chacauhaa (chacau means warm and haa means cold). This word
became mixed with the ancient word cacauatl, giving rise to the word chocolate. The word
chocolate is used for the industrialized final product, which possesses the taste and smell
typical of the product.
Each fruit pod contains 30–40 beans embedded in a mucilaginous pulp. Raw cocoa has
an astringent, unpleasant taste and flavor and has to be fermented, dried, and roasted to obtain
the characteristic flavor and taste of cocoa (Nielsen et al., 2007). Cocoa fermentation is one of
the stages during post-harvest processing that ultimately governs the product quality.
Fermentation remains empirical and does not give rise to beans of consistent quality, which
obliges processors to make continuously changes to their formulations (Gálvez et al., 2007).
The fermentation of cocoa is a spontaneous microbiological process. The interior of the
unopened pods are considered to be sterile or almost sterile, but following opening of the
pods the cocoa beans are contaminated with a variety of microorganisms originating from,
e.g., workers’ hands, containers for transport, knives, pod surfaces, and other sources (Nielsen
et al., 2007). Fermentation helps to break down the mucilaginous pulp surrounding the beans
and causes cotyledon death. It also helps to trigger biochemical changes inside the beans that
contribute to reducing the bitterness and astringency, as well as to the development of flavor
precursors (Gálvez et al., 2007). Several different fermentation systems are used around the
world, with heap and box fermentations being the most widely used (Nielsen et al., 2007).
Following harvest, the pods are broken open and the beans are piled on and covered with
plantain leaves. Aeration of the heaps by turning them every 24–72 h during fermentation is
considered beneficial for the quality of the end product, although this strategy is not utilized
by all farmers (Baker et al., 1994; Nielsen et al., 2007).
Early on in cocoa fermentation, several species of yeasts proliferate, leading to the
production of ethanol and the secretion of pectinolytic enzymes. This is followed by a phase
during which bacteria appear, principally lactic-acid bacteria and acetic-acid bacteria,
followed by growth of aerobic spore-forming bacteria. Finally, some fungi may appear on the
surface of the fermentation (Schwan and Wheals, 2004). During the first phase of
fermentation, the yeast possess an intense metabolism that is favored by the acidity of the
environment, the richness of fermentable carbohydrates, and the low oxygen content of the
mass (Lehrian and Patterson, 1984; Schwan and Wheals, 2004). Yeast fermentation
metabolism very quickly leads to consumption of all the simple sugars, giving rise to ethanol
25
Fátima C. O. Gomes, Inayara C. A. Lacerda, Diego Libkind, et al.
and carbon dioxide. The pectinolytic activities of the yeasts causes degradation of the pulp,
increasing the aeration of the fermenting mass, and the assimilation of citric acid by yeasts
and lactic acid bacteria causes the pH to rise (Thompson et al., 2001). This in turn favors the
growth of aerobic acetic acid bacteria. These bacteria metabolize the ethanol initially formed
by the yeasts to acetic acid through an exothermal process. Ethanol and acetic acid diffuse
into the beans. This, in combination with the heat produced by the activities of the acetic acid
bacteria, leads to bean death, killing the embryo, and induces biochemical changes leading to
well-fermented cocoa beans (Thompson et al., 2001). Due to the activity of the acetic acid
bacteria, the temperature of the fermenting mass increases to 45–50°C, thereby limiting the
growth of many microorganisms. Increased aeration, an increased pH value (3.5 to 5.0) of the
cocoa pulp, and a rise in temperature to about 45◦C in the cocoa mass during later stages of
fermentation are associated with the development of aerobic spore-forming bacteria of the
genus Bacillus. Many Bacillus spp. are thermotolerant, and others grow well at elevated
temperatures. Aerobic spore-forming bacteria produce a variety of chemical compounds
under fermentative conditions. These may contribute to the acidity and perhaps sometimes to
the off-flavors associated with fermented cocoa beans (Nielsen et al., 2007).
The traditional drying of fermented cocoa beans is carried out in the sun (Oetterer et al.,
2006). During the drying process, the enzymes present in the bean promote the chemical
reaction of aging, producing the characteristic flavor and color of chocolate. The temperature
of drying is important for the final quality of the cocoa beans. The best temperatures range is
between 35 and 40oC, which are optimal temperatures for enzyme activity (Oetterer et al.,
2006). However, to improve the quality of the processed beans, more research is needed on
pectinase production by yeasts, better depulping, fermenter design, and the use of starter
cultures for cocoa bean fermentation (Schwan and Wheals, 2004).
3.4. Coffee
Brazil is the largest producer of coffee (4.2Mtons), followed by Paraguay, Venezuela,
Colombia, Indonesia, Ethiopia, India, Mexico, and another 40 countries (Silva et al., 2008).
Two coffee species dominate the world market: Coffea arabica (arabica) and C. canephora
(robusta). Arabica and Robusta coffees account for 76.4% and 23.6% of production
worldwide, respectively (Coltro et al., 2006; Silva et al., 2008). The amounts of chemical
compounds in the coffee beans undergo variations during the development and maturation of
the fruit, until they reach the correct levels characteristic of the ripe coffee.
Coffee’s commercial quality is associated with a set of factors that involve physical-
chemical and sensorial aspects, which, in turn, depend on the products’ drying and storage
conditions (Afonso Júnior, 2001; Nobre, 2005). Several techniques are used to dry coffee
cherries, mainly on the ground. During the drying process, variations in the structure of the
beans (color, aspect, defects, bouquet, and flavor) can occur, affecting the quality of the
beverage. External factors, such as temperature, relative humidity, and mechanical damage
can alter the structure of the membranes, leading to a loss of their organization and selectivity
(Amorim et al., 1978) and reducing the coffee’s quality potential. Corrêa et al. (1994), Sfredo
et al. (2005), and Souza (2000) investigated coffee drying and concluded that in order to
obtain a good quality product and a soft beverage, the drying temperature must be maintained
around 40º C. However, to preserve the coffee’s initial qualities, correct drying is not enough.
26
Traditional Foods and Beverages from South America: …
Adequate storage is also important. To store coffee for longer periods of time and maintain its
initial chemical, physical, and sensorial characteristics, the temperature and relative humidity
of the environment must be monitored. Afonso Júnior (2001) observed a decrease in the
composition of reducing sugars in coffee beans and grains as the storage time in higher
relative humidity conditions increased.
Coffee cherries are processed using one of the two methods (Schwan and Wheals, 2003).
In Colombia, Central America, and Hawaii, the ‘wet’ method is used for Arabica coffee. In
the wet method, hand-picking of mature cherries is followed by mechanical depulping and
then fermentation for approximately 24–48h to remove the mucilage layer. The dry process,
which results in so-called unwashed or natural coffee, is the oldest and simplest method used
to process coffee. The dry process is often used in countries where rainfall is scarce and long
periods of sunshine are available to dry the coffee properly. The dry method is used for about
95% of the Arabica coffee produced in Brazil, most coffee produced in Ethiopia, Haiti,
Indonesia, and Paraguay, and some Arabica produced in India and Ecuador (Silva et al.,
2008). This method involves the fermentation of whole fruit and usually produces coffee that
is heavy in body, sweet, smooth, and complex. The coffee fruits are spread on the ground
(earth, platforms, concrete, or asphalt) in layers approximately 10cm thick, heaped at night,
and respread each day (Schwan and Wheals, 2003). Over the course of 10–25 days of sun
drying, natural microbial fermentation occurs that can influence the final quality of the
product (Schwan and Wheals, 2003; Silva et al., 2000). The fermentation of pectinaceous
sugars produces ethanol and acetic, lactic, butyric, and higher carboxylicacids. The formation
of butyric and propionic acids from bacterial fermentation causes a loss of quality due to the
diffusion of the acids into the fruit cherries (Amorim and Amorim, 1977). Bacteria, yeasts,
and filamentous fungi have previously been reported during fermentation by the ‘wet’ method
(Masoud and Kaltoft, 2006; Avallone et al., 2001; Silva et al., 2008), but only one
comprehensive study of dry processing has been published (Silva et al., 2008). The
microbiota involved in ‘dry’ processing are much more varied and complex than those found
during wet fermentation, although the actual role of each group of microorganisms during
coffee fermentation by natural processing remains unknown. The microbial succession and
consortium of bacteria, yeast, and filamentous fungi and their metabolites during natural
coffee fermentation remain to be studied. An understanding of microbial dynamics during
natural fermentation should enable more rapid fermentation and better quality of the final
product. The constant demand of the consumer market for high-quality coffees has led to the
need to understand the total microbial environment of dry or naturally processed coffee, and
to determine the organoleptic characteristics of the final beverage (Silva et al., 2008).
4. CONCLUDING REMARKS
Many traditional fermented beverages and foods from South America are finding a
market throughout of the world; however, some of these items are characterized by only local
production and exhibit several technological problems. The lack of knowledge regarding the
microbial communities associated with fermentation is a key problem for improving the
manufacture of these products. Some products, such as wine, cachaça, and traditional cheeses,
have been studied in more detail in recent years, and they possess greater flavor quality than
27
Fátima C. O. Gomes, Inayara C. A. Lacerda, Diego Libkind, et al.
those products fermented using ancient techniques. Research to elucidate new starter
microbial cultures and new technologies for the production of these products are imparting
sensorial sophistication to these traditional products. The market expansion of these products
is dependent on the improvement of the technological strategies used to produce these
traditional fermented foods and beverages.
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... Other theories suggest that the name is derived from the word Chibcha, a civilization that populated Colombia and Panama, or relate the word chicha to Chichas, an ethnicity present in southern Bolivia before the establishment of the Incas. 1 Chicha is a clear, yellow, and frothy beverage present in the Andean region and in low-lying regions of Ecuador, Peru, Bolivia, Colombia, Brazil, and Argentina. 2 This traditional beverage is prepared mainly from maize, but currently, the name is considered generic and refers to a variety of beverages, fermented or not, prepared from various other materials, such as cassava, beans (such as rice, oats, and quinoa) and fruits (such as bananas). ...
... In Ecuador, the first reports of chicha production date back to 200 B.C., before the establishment of the Incas in the region. 1 This beverage was of great importance in traditional indigenous cultures, especially in the Incan culture, wherein it was also linked to festive ceremonies. 3 In Ecuador, as in the rest of the Andean region, the most common maize chicha is chicha de jora (Fig. 1). ...
... The non-Saccharomyces species found in our study can be isolated from different substrates worldwide, including those of fermentation processes for beverage production. 1,5,[20][21][22] The origin of the non-Saccharomyces species associated with the chicha samples studied might be explained by the different manufacturing processes for these beverages. Chicha de jora is prepared in different ways, employing a wide variety of raw materials with additional ingredients such as fruits, herbs, spices, brown sugar, and sugar. ...
Article
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Chicha, a type of beer made mainly with maize or cassava, is a traditional fermented beverage of the Andean region. There have only been a few studies on yeasts associated with chicha fermentation, and the species diversity occurring during the production of this beverage is not known. The objective of this study was to determine the biodiversity of yeasts in chicha, and to characterize the Saccharomyces cerevisiae populations associated with the production of chicha de jora, seven-grain chicha, chicha de yuca, and chicha de morocho in Ecuador. The molecular diversity of S. cerevisiae populations was determined by restriction polymorphism mitochondrial DNA (mtDNA) profiles. The beverages were characterized based on their physicochemical parameters. Twenty-six species were identified, and the most prevalent species were S. cerevisiae and Torulaspora delbrueckii. Other yeast species were isolated at low frequencies. Among 121 isolates of S. cerevisiae, 68 different mtDNA molecular profiles were identified. These results showed that chichas are fermented by a high number of different strains of S. cerevisiae. Some other species provided a minor contribution to the fermentation process. The chicha presented generally similar physicochemical parameters to those observed for other traditional fermented beverages, and can be considered as an acid fermented beverage.
... The ancestral strategy to prepare these beverages usually included steps of chewing and/or germination of corn, but currently some modifications have occurred in both the form of preparation and used ingredients. The most important and common steps for preparing Ecuadorian chicha are boiling, cooling and fermentation (Gomes et al., 2009). Sugar, panela (non-centrifugal sugar cane), as well as herbs and spices, can be added (Faria-Oliveira et al., 2015). ...
... Although we found only six S. cerevisiae strains and two T. delbrueckii strains in the analysed Ecuadorian chichas, they showed physiological differences between strains, as reflected in particular stress tolerance profiles. A combination of two events, domestication and dispersion, is probably the origin of chicha yeasts (Gomes et al., 2009). So the variability of our Ecuadorian isolates could be a consequence of them. ...
Article
The potential of yeasts isolated from traditional chichas as starter cultures, either for controlled production of the native beverage or for industrial beer production, has been investigated. Three S. cerevisiae strains and one T. delbrueckii strain isolated from four different Ecuadorian chichas were compared to ale and lager beer strains with respect to fermentation performance, sugar utilisation, phenolic off-flavour production, flocculation and growth at low temperature. Fermentations were performed in 15 °P all-malt wort and in a model chicha substrate at 12 °C and 20 °C. Tall-tube fermentations (1.5 L) were also performed with both substrates to assess yeast performance and beer quality. Among the strains tested, only one Ecuadorian S. cerevisiae strain was able to ferment the wort sugars maltose and maltotriose. Fermentations with all Ecuadorian strains were poor in wort at 12 °C relative to 20 °C, but were similar in model chicha substrate at both temperatures. The aromatic profile was different between species and strains. These results indicate the potential of yeasts derived from traditional Andean fermented beverages for commercial applications. One of the chicha strains demonstrated traits typical of domesticated brewery strains and could be suitable for ale fermentation, while the other strains may have potential for low-alcohol beer or chicha production.
... The ancestral strategy to prepare these beverages usually included steps of chewing and/or germination of corn, but currently some modifications have occurred in both the form of preparation and used ingredients. The most important and common steps for preparing Ecuadorian chicha are boiling, cooling and fermentation (Gomes et al., 2009). Sugar, panela (non-centrifugal sugar cane), as well as herbs and spices, can be added (Faria-Oliveira et al., 2015). ...
... Although we found only six S. cerevisiae strains and two T. delbrueckii strains in the analysed Ecuadorian chichas, they showed physiological differences between strains, as reflected in particular stress tolerance profiles. A combination of two events, domestication and dispersion, is probably the origin of chicha yeasts (Gomes et al., 2009). So the variability of our Ecuadorian isolates could be a consequence of them. ...
Article
Yeasts involved in the spontaneous fermentation of traditional beverages like chicha (indigenous Andean beer) may have the potential to be used as starter cultures to improve the quality and microbiological safety of these products, but also as non-conventional alternatives to other food alcoholic fermentations. In this research, we isolated, identified and characterised yeast strains from four Ecuadorian chichas made by using four different raw materials: rice (RC), oat (OC), grape (GC) and a mixture of seven corn varieties (yamor, YC). Finally, 254 yeast isolates were obtained and identified by molecular methods. Eleven yeast genera and 16 yeast species were identified with relatively few isolates belonging to Saccharomyces cerevisiae (9.1% belonging to 6 strains) and Torulaspora delbrueckii (18.6% belonging to 2 strains). In order to select good candidates for fermentative starter production, different analyses were performed. The results of the stress response tests showed a wide variability between species and strains, and identified some yeasts displaying high stress tolerance, similarly to commercial wine strains. Amylase production was screened as being indicative of the capacity to degrade and ferment starch-rich substrates. A Cryptococcus sp. isolate showed the highest amylase activity. The growth rate and fermentative capacity in molasses medium was measured for three S. cerevisiae, T. delbrueckii and Candida sp. strains as tests for yield and performance in biomass production. Based on their excellent behaviour, three S. cerevisiae strains and one T. delbrueckii strain were selected for further analyses, including dehydration tolerance and invertase activity as additional desired traits for chicha starters. All the S. cerevisiae strains exhibited high invertase activity and one also displayed high resistance to dehydration. The yeasts selected in this study can thus be suitably used as dry starters for the microbiologically controlled production of traditional beverages, and also for other alcoholic fermentations.
... Both yeast species are ubiquitous being found in a wide range of latitudes, including in Antarctica glaciers (data not published). Some registers of R. glutinis are in atmosphere, trees, leaves, grapes, soil, spoiled leather, sea water, water supply of a brewery, sputum of pneumonia patient, exudates, limph nodes, feces, pasteurized beer and a number of other substrates [41]. These two yeast species are not fermentative, which is a trait of basidiomycetous yeast species. ...
... This yeast species has been found in flowers, rotten wood, insects, excrement, rotten vegetal matter, and fungus ( Figure 3a). Candida tropicalis has been reported to grow even at pH so high as 10 [41], which is a remarkable feature in terms of tolerance facing unusually hard environmental conditions. ...
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In terms of the substrates where the yeast species have been found both in mainland and the islands there is a wide range of sources: insects, flowers, leaves, feces, sugar cane mills, fungus, fruit, moss, and a number of samples taken from endemic plants in the Galápagos Islands, including Miconia robinsoniana, Scalesia sp., Opuntia sp. Castela galapageia, etc. This genus is widely represented in the CLQCA, where about 100 isolates were collected from all the regions in mainland and Galápagos. The isolates from Galápagos represent about 60% of the total isolates of this genus in the CLQCA. C. carvajalis [43] was the first yeast species described from Ecuador. This yeast was found in the course of a yeast biodiversity survey in the Amazonia. The substrate sampled was rotten wood and fallen leaf debris, collected around crude oil wells, close to Dayuma town. One Isolate of this species was collected in Santa Cruz Island from Psidium guajava mucilage. The closest relatives of C. carvajalis are C. asparagi, C. fructus, and C. musae. This group of yeasts belongs to the Clavispora clade. In Mainland Ecuador it has not collected any C. asparagi [44] isolate, but in the Galápagos Islands we have one register from Santa Cruz Island where this species was collected from a nitidulid beetle. In this chapter we have developed a new ecological approach by means of a mathematical model which is useful for a better understanding of the adaptability of yeasts as well as the specialization degree of these microorganisms in Ecuadorian ecosystems. The data herein processed will be completed in future expeditions, but constitutes a base for the upcoming ecological studies of the yeasts in the Galápagos Islands and Ecuadorian Mainland. The mathematical model shows an inverse correlation between the “Index of Specialization” (Si) and the “Index of Abundance” (Ia). Moreover, it can be seen that the trend is towards the specialization since 70 out of 104 yeasts species analyzed (c.a. 67%) showed a Si between 0.92 to 0.53, which means that they were isolated from a maximum of three out of seven ecosystems and a maximum of three out of nine substrates; 30 yeast species (c.a. 29%) showed an intermediate Si, between 0.18 and 0.47, meaning that these yeasts species were found in a maximum of six different ecosystems and six different substrates; finally, only four yeast species (c.a. 4%) showed a very low Si, between 0.02 and 0.11, which means that these species were found in up to seven ecosystems and nine substrates analyzed. These four yeast species are considered the more generalist and exhibit the highest adaptability, but represents a minority in the complete pool of yeast species studied. The total number of ecosystems analyzed was seven and the total number of substrates studied were 10. No yeast species were found in all the 10 substrates.
... For example, laboratory analyses (Almeida et al., 2007;Colehour et al., 2014;L. Ramos, 2013;Gomes et al., 2009) revealed that traditional fermented beverages in many cases were not alcoholic or had a low alcohol concentration. This confirms what many chroniclers and anthropologists wrote, that fermented beverages were primarily a food and not a drink meant to inebriate (Barghini, 2018). ...
Article
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In South America’s lowlands, it was believed that fermentation processes for amylaceous substances were performed only with the inoculation of salivary amylase in the mash. In 2004, Henkel identified a unique fermentation process that was not previously known in the Americas: fermentation performed with the inoculation of an amylolytic mold, Rhizopus sp. Amylolytic fermentation is an important way to transform and enrich carbohydrates and is widely used in the eastern and southeastern Asia for the enhancement of beverages and food. To verify if this process was unique, extensive research has been carried out by analyzing reports by missionaries, travelers, and anthropologists to search for hints of a larger diffusion of such processes. This research made it possible to verify that the use of amylolytic molds was widespread throughout the Amazon basin, from Rio Napo and the upper Amazon to Guianas and Orinoco. In addition, it was possible to verify that Rhizopus sp. was not the only mold employed. It is probable that the other molds used were Monascus sp. and Aspergillus sp. This leads us to believe that the fermentation processes in the Amazon basin were likely more varied than previously thought, and are worthy of deeper studies.
... Some indigenous groups in southern South America, particularly the Mapuche, had a tradition of making a fermented beverage called chichi from Cyttaria (Schmeda-Hirschmann et al. 1999;Gomes et al. 2010). This traditional use led researchers to investigate sugar levels and the diversity of yeasts associated with Cyttaria species. ...
... Although the recipes pass from generation to generation, all of them use the conversion of starch into sugar, followed by fermentation of sweet wort. As the production process resembles the brewing process, the traditional chicha, made from maize, can be named as the Andean indigenous beer [67]. ...
Chapter
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Fermentation is one of the oldest forms of food preservation in the world. In South America, most fermented beverages are non-dairy products featuring several other food raw-materials such as cereals, fruits and vegetables. Generally, natural fermentations are carried out by yeast and lactic acid bacteria forming a complex microbiota that acts in cooperation. Yeast has a prominent role in the production of beverages, due to the ability to accumulate high levels of ethanol and to produce highly desirable aroma compounds, but lactic acid bacteria are particularly important in fermentation because they produce desirable acids, flavour compounds and peptides that inhibit the growth of undesirable organisms. Amongst the South America beverages based on cereals and vegetables, could be cited the fermented beverages chicha, caxiri, cauim and champús, and cachaça, a fermented and distilled beverage. Genetic and physiological analyses of S. cerevisiae strains isolated from cachaça have been shown to present interesting traits for beer production, such as flocculation and production of aroma compounds, fundamental to high quality beer. The study of these traditional beverages allows the identification of new microorganism strains displaying enhanced resistance or new flavour and aroma profiles that could lead to applications in several industries and ultimately new products
Article
The development of early civilizations was greatly associated with populations’ ability to exploit natural resources. The development of methods for food preservation was one of the pillars for the economy of early societies. In Ecuador, food fermentation significantly contributed to social advances and fermented foods were considered exclusive to the elite or for religious ceremonies. With the advancement of the scientific research on bioprocesses, together with the implementation of novel sequencing tools for the accurate identification of microorganisms, potential health benefits and the formation of flavor and aroma compounds in fermented foods are progressively being described. This review focuses on describing traditional fermented foods from Ecuador, including cacao and coffee as well as less popular fermented foods. It is important to provide new knowledge associated with nutritional and health benefits of the traditional fermented foods.
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La región nor-patagónica (principalmente las provincias de Río Negro y Neuquén) cuenta con una gran tradición en la industria de las fermentaciones de diversos productos como son vino, sidra y cerveza. La producción de cerveza en la región es de índole artesanal y en los últimos años ha experimentado un crecimiento sin precedentes que está en línea con lo observado en otras regiones del país y de Sudamérica (ej. Chile y Brasil). El descubrimiento en la Patagonia Andina de una nueva especie de levadura Saccharomyces eubayanus, parental del híbrido inter-específico S. pastorianus (utilizado a nivel mundial en la producción de cerveza Lager), abrió un campo sumamente fértil para la investigación, desarrollo e innovación. Este trabajo busca contribuir al conocimiento sobre la biogeografía de S. eubayanus y la estructura genética de sus poblaciones, así como caracterizar y desarrollar cepas con potencial aplicación a la industria cervecera. Para expandir el conocimiento de la distribución de S. eubayanus en la Patagonia Andina, se aumentó cinco veces el área de estudio, y se incorporaron nuevas especies de árboles nativos y exóticos. A partir del análisis de 563 muestras se detectaron porcentajes muy elevados de incidencia de levaduras (70 %), de levaduras sacaromicéticas (54 %) y en particular de S. eubayanus (31 %) en comparación con datos de otros lugares del mundo. Se obtuvieron e identificaron 175 aislamientos naturales de la Patagonia Argentina y Chile a los que inicialmente se les secuenciaron un marcador molecular mitocondrial (COX2) y dos nucleares (DCR1, intFR) para obtener los primeros datos de la estructura poblacional de la especie en la región. El gen mitocondrial COX2 permitió determinar aislamiento por distancia hacia el sur de la Patagonia Andina y encontrar casos de eventos de evolución reticulada (introgresiones) con su especie simpátrica y hermana S. uvarum. Los marcadores nucleares permitieron detectar dos grandes poblaciones de S. eubayanus: una, denominada PB/Hol, establecida a lo largo de toda la Patagonia Andina y filogenéticamente cercana a los aislamientos Holárticos, la otra restringida al Norte de la Patagonia (PA). El marcador intFR fue útil para la determinación de 5 sub-poblaciones, 2 de PA y 3 PB, una de ellas no descripta previamente y asociada exclusivamente a la región sur de la Patagonia. Se observó una estructuración genética que sugiere la presencia de antiguas barreras al flujo genético en el paralelo 43°, que se correlacionan con fenómenos equivalentes detectados para especies vegetales, en particular Nothofagus. Además se obtuvieron borradores genómicos de 127 nuevos aislamientos de S. eubayanus a los que se los estudió mediante Single Nucleotide Polimorphism (SNPs), lo que permitió no solo verificar las 5 sub-poblaciones, sino que también detectar linajes aún más divergentes, y observar una estructuración filogeográfica inédita para las especies del género. Ésto, así como la gran diversidad y abundancia de S. eubayanus en la Patagonia, refuerzan la hipótesis de que dicha especie es nativa de la región. Dado que hasta el momento solo se contaba con datos limitados de las propiedades fermentativas de la especie y únicamente de la cepa CRUB1568T, se procedió a obtener datos complementarios para dicha cepa, en particular sobre su comportamiento en condiciones industriales de fermentación en altas densidades (High Gravity Brewing, HGB). Además, se hizo uso de la inédita diversidad genética disponible para la especie, y se realizó un relevamiento de las propiedades fermentativas de 60 aislamientos, incluyendo representantes de las distintas sub-poblaciones detectadas. Se seleccionó un representante de cada sub-población, con características fermentativas diferenciales, y se evaluó su desempeño en mosto cervecero. De las cinco sub-poblaciones analizadas, cuatro lograron una fermentación de maltosa casi completa (equivalente a cepas comerciales), aunque ninguna logró metabolizar la maltotriosa teniendo como resultado cervezas con un 65 % de atenuación (siendo 85 % la atenuación promedio para cepas de tipo Lager). Para todas las cepas estudiadas, se detectaron importantes compuestos de sabor y aroma (flavor) pertenecientes al grupo de los fenoles volátiles, como el 4-vinil guiaicol y 4-vinil fenol, que aportan a las cervezas producidas características organolépticas especiales, sólo compartidas con levaduras comerciales para elaborar cervezas belgas y de trigo. En este trabajo se generó un primer acercamiento a la genómica comparativa de S. eubayanus haciendo foco en 33 genes de interés relevantes para la producción de cerveza. Se determinó que la cepa tipo CRUB1568T es una las que posee las mejores propiedades fermentativas de las poblaciones patagónicas relevadas. Sin embargo, características como la baja atenuación, floculación y velocidad de fermentación condicionan y limitan su transferencia al sector productivo. Por esto, a partir de S. eubayanus CRUB1568T se buscó desarrollar cepas con propiedades fermentativas mejoradas y/o diferenciales por dos métodos no generadores de organismos genéticamente modificados (OGM): evolución experimental dirigida e hibridación inter – específica. El primer método permitió la obtención, luego de 430 generaciones, de una cepa (A62) que presentó mejoras significativas respecto de su parental en los parámetros: velocidad de fermentación y atenuación. El segundo método, permitió la obtención de 9 híbridos inter-específicos con el método espora – espora con cepas de levaduras cerveceras de tipo Ale con capacidad esporulante. Lamentablemente las mismas no evidenciaron vigor híbrido en las variables de cinética de fermentación analizadas, aunque se detectaron preliminarmente diferencias organolépticas. Por ello, la cepa evolucionada A62 fue seleccionada para los estudios de escalado posteriores y para un test de preferencia público. El estudio, realizado con 232 participantes, permitió determinar que la cepa evolucionada es diferente organolépticamente de la cepa original (CRUB1568T) y que estas diferencias no reducen la aceptabilidad del público. Se encuentra en trámite la gestión del patentamiento de la cepa evolucionada. El trabajo realizado en el marco de esta tesis es el más exhaustivo del mundo en lo que refiere al aislamiento y caracterización fenotípica, genética y genómica de S. eubayanus. Se aprovechó esta información para seleccionar las cepas salvajes de mayor interés para innovación en la industria cervecera, y empleando técnicas no generadoras de OGM se desarrolló una cepa ampliamente mejorada en velocidad de fermentación y atenuación, con alto potencial de transferencia al sector productivo.
Chapter
A large variety of fermented foods and beverages with traditional and cultural value have been described in the world, including industrial products such as wine, cider, and beer as well as traditional ones. In contrast with the massive scientific information available about the microbiota responsible for winemaking, yeasts responsible for most traditional fermented beverages around the world remain undiscovered. Both industrial and traditional fermentation processes coexist in Patagonia, making this region an ideal scenario for fermentative yeast diversity studies. The most relevant feature of this area is the fact that most traditional processes are produced at low temperature (below 20 °C), which directly affects the microbial diversity. We identified and characterized fermentative yeasts present during industrial fermentations of wine and cider and traditional fermentations (chichas) obtained from wild apples and Araucaria araucana seeds, substrates typically used by aboriginal communities to prepare soft alcoholic beverages. As a general rule, only Saccharomyces cerevisiae strains were obtained from wines and ciders, and they showed a close genetic relationship with European strains of this species. In traditional fermentations, commercial bakery and European wine strains of S. cerevisiae were detected as pure or mixed cultures with Saccharomyces uvarum, a cryotolerant species. This last species was also isolated from A. araucana seeds in Patagonian forests together with Saccharomyces eubayanus, another cryotolerant species of the genus. Genetic information obtained from the analysis of S. uvarum from apple chichas evidenced a closer relationship to industrial (European) strains than to natural (Patagonian) strains of this species. North Patagonia is an interesting scenario to study cryotolerant (S. uvarum and S. eubayanus) yeast diversity studies, and a source of new strains with potential biotechnological interest.
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The objective of the work was to evaluate alterations in the quality of natural and washed coffee under different drying conditions (coffee drying yard, temperature of 40ºC and 60ºC) and storage conditions at 60% of relative humidity, with controlled temperature of 23ºC, at 90 and 180 days. The work was carried out in the Engineering Department and in the Coffee Post-Harvest Technology Pole of the Federal University of Lavras. The manual harvest of the coffee (Coffea arabica L.), Topázio variety, was selective. Part of the coffee was pulped and the other part was processed in the natural form. A portion of each type of coffee was submitted to drying on the yard and two other samples were processed in a mechanical dryer, at temperatures of 40ºC and 60ºC. After drying, the coffee was stored in an air-tight room, in which a stable relative humidity of 60% was maintained with a solution of magnesium nitrate. Quality evaluation, sensorial analyses, electric conductivity and potassium leaching tests, total titrable acidity, fatty acidity and total and reducing sugars determinations were carried out. The results showed that the coffee dried at 60ºC, after 90 days storage, presented the poorest quality. The physical-chemical evaluations of the drying and storage conditions showed that washed coffee presents better quality when compared to the product in its natural form.
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The biological activity and nutritional composition of Chilean collections ofCyttaria berteroi C. darwinii, C. espinosae, C. harioti andC. johowii have been determined. The crude protein, lipid, ash, and carbohydrate content of the samples examined were similar to that of other edible fungi. Amino acid analysis of ChileanCyttaria showed that proteins of all species are deficient in methionine and cysteine and excepting oneC. espinosae collection, all samples proved to be below the WHO values for the essential amino acids valine, isoleucine, leucine, and lysine. The acute oral toxicity test ofC. espinosae in rats showed that doses up to 2.5 g extract/kg body weight (corresponding to 25.7–38.7 g fresh weight/kg) did not produce mortality or macroscopic damage in the organs examined of the test animals. Cyttaria extracts assayed at 50 fig/ml were inactive or moderately active as inhibitors towards the enzymes xanthine oxidase (0-31%) and β-glucuronidase (0-65%), and lacked antifungal and antibacterial effects in a battery of antimicrobial assays. When administered intravenously at 2.5 mg/kg, the water-soluble extract ofCyttaria produced a hypotensive response in rats (-16 to -21%). Furthermore, most of the aqueous extracts ofC. espinosae andC. harioti showed DNA binding activity. The main sterols fromCyttaria espinosae andC. berteroi were identified as dihydrobrassicasterol derivatives. Our study suggests that edibleCyttaria species do not represent an acute toxicity risk for consumers and that their nutritional value is similar to that of other edible, cultivated mushrooms. The preservation ofCyttaria spp. as a food resource is linked to the protection of the temperateNothofagus forests.
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The aim of this study was the survey of killer yeasts in natural environments at the Nahuel Huapi National Park in the Subantartic Forest, Northwestern Patagonia. One hundred and twenty three yeast isolates were analysed. Yeasts associated with nectarine flowers, sugary wild fruits and sporangia of the fungus Cyttaria spp., and glacial-origin lakes freshwater samples were screened in their killing activity against the sensitive yeast tester strain Candida glabrata NCYC 388. The sensitivity to known toxins (K 1 to K 10) was tested against yeast isolates. Three of the 28 cultures isolated from nectar showed killer activity, whereas none was detected in those derived from fruits or freshwater samples. The 38% of all tested strains were sensitive to one or more toxins. The broadest spectrum of sensitivity was observed in cultures from nectar samples, while 57% of the yeasts isolated from fruits were neutral. The latter were also prevalent among yeasts from freshwater samples.
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The dynamics of the wine yeast strains presented in five spontaneous Malvasia wine fermentations have been studied. Samples were analysed for their microbiological characteristics and chemical substances. All 937 isolates were characterized using electrophoretic karyotyping and tested for their killer activity. The non- Saccharomyces population was identified using a combination of PCR-RFLP analysis of the rDNA spacer region and physiological testing. The total yeast population level in the must after sedimentation was 105cfu ml−1and included the following genera:Candida, Metschnikowia, Hanseniaspora, Rhodotorula, Issatchenkia and Debaryomyces. However, Saccharomyces sp. was not detected in fresh must samples plated on YEPD medium. Based on the chromosome length polymorphism among 649 isolates from the subsequent phases of fermentation, 46 different electrophoretic patterns of Saccharomyces cerevisiae were distinguished. The most abundant karyotypes were L1, L4, L12, P6. A sequential substitution of S. cerevisiae strains occurred during the different phases of fermentation. At the slow fermentation rate, karyotype L4was most abundant in almost all fermenters. At the beginning of the tumultuous fermentation phase, the most frequent karyotype became L1followed by karyotype L4. Finally, during the fermentation process, pattern L4was clearly replaced by karyotype L1followed by pattern L12. Despite the same fermentation source (grape must), differences among five spontaneous fermentations were observed. The population dynamics of S. cerevisiae yeasts, especially the dynamics of the major S. cerevisiae strains (L1, L4, and L12) were quite similar in all five fermenters in opposite to the minor strains of S. cerevisiae.
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In Brazil and Colombia, ‘sour starch’ is traditionally obtained by a submerged lactic fermentation of crude cassava starch followed by sun drying. It is used by local bakers to prepare breadlike products which display the same expanded crumb texture as in wheat bread. In this process, suspended starch is settled down and left aside for a few weeks under anaerobic conditions where natural lactic populations develop. Three collections of clones isolated from local fermentations have been identified using the API procedure and further characterized. Most of them belong to different species of Lactobacillus. Many display a ropy phenotype, typical for exopolysaccharide (EPS) excretion. A possible role of these EPS in the special properties of sour starch is discussed.