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MICROORGANISMS

IMPORTANT IN FOOD
MICROBIOLOGY:

Molds, Yeasts and


Yeastlike Fungi,
Bacteria
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MOLDS

GENERAL CHARACTERISTICS OF MOLDS

Molds are multi-cellular, filamentous, branched. Filamentous


fungi whose growth on foods usually is readily recognized by
fuzzy or cottony appearance. The main part of the growth
commonly appears white but may be coloured or dark or
smoky.

Coloured spores are typical of mature mold of some kinds


and give colour to part or all of the growth.

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Identified by physical appearance, colony characteristics,
and reproductive spores.
Thallus: vegetative body, is characteristic of thallophytes,
which lack true roots, stems, and leaves. Body of a mold or
fleshy fungus. Consists of many hyphae.
Hyphae (Sing: Hypha): Long filaments of cells joined together.
Septate hyphae: Cells are divided by cross-walls (septa).
Coenocytic (Aseptate) hyphae: Long, continuous cells that
are not divided by septa.
Hyphae grow by elongating at the tips.
Each part of a hypha is capable of growth.
Vegetative Hypha: Portion that obtains nutrients.
Reproductive or Aerial Hypha: Portion connected with
reproduction.
Mycelium: Large, visible, filamentous mass made up of many
hyphae.
Molds produce sclerotia (singular sclerotium),
which are tightly packed masses of modified
hyphae, often thick-walled, within the mycelium.
Sclerotia are considerably more resistant to heat
and other adverse conditions than is the rest of
the mycelium.

Molds are divided into two groups: septate, with


cross walls dividing the hyphae into cell and
noncoenocytic, septate with the hyphae
apparently consisting of cylinders without cross
walls.

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Characteristics of Fungal Hyphae:
Septate versus Coenocytic
Mycelium: Large, Visible Mass of
Hyphae
Diagram illustrating the ultrastructure of a septate hypha

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The Fungal Wall
Functions :
PROTECTS the underlying protoplasm;
determines and MAINTAINS THE SHAPE of the fungal cell
or hypha;
acts as an INTERFACE between the fungus and its
environment;
acts as a BINDING SITE for some enzymes;
possesses ANTIGENIC properties - which allow interactions
with other organisms.

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Chemical composition of the wall:
POLYMERIC FIBRILS
chitin
cellulose (in the Oomycota)

AMORPHOUS MATRIX COMPONENTS


glucans
proteins
lipids
heteropolymers (mixed polymers) of mannose, galactose,
fucose and xylose

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Reproductive parts or structures
Mold can grow from transplanted piece of mycelium.
Reproduction: asexual spores, some molds from sexual
spores.
perfect, classified as Oomycetes or Zygomycetes if
non septate, or Ascomycetes or Basidiomycetes if
septate.
imperfect, the fungi Imperfecti (typically septate),
which have only asexual spores.

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LIFE CYCLE OF FUNGI
Filamentous fungi can reproduce asexually by fragmentation
of their hyphae.
Fungal spores are formed from aerial hyphae and are
used for both sexual and asexual reproduction.
1. Asexual spores: Formed by the aerial hyphae of one organism. New
organisms are identical to parent.
Conidiospore: Unicellular or multicellular spore that is not
enclosed in a sac.
Chlamydospore: Thick-walled spore formed within a hyphae
segment.
Sporangiospore: Asexual spore formed within a sac (sporangium).
2. Sexual spores: Formed by the fusion of nuclei from two opposite
mating strains of the same species. New organisms are different from
both parents.
Asexual spores
Produced in large numbers and are small, light, and
resistant to drying.
The four principal types of asexual spores are:
1. Conidia (singular conidium)
2. Arthrospores, or oidia (singular oidium)
3. Sporangiospores
4. Chlamydospore
The swollen tip of the sporangiophore, the columella,
usually projects sporangium.
Some molds have conidia in chains, squeezed off one
by
one from a special cell, a sterigma (plural sterigmata)
or 12
phialide, at the tip of the Conidiophore.
Sexual spores
The nonseptate molds (Phycomycetes) that produce
oospores are termed Oomycetes. These molds are
mostly aquatic, group are several important plant
pathogens, downy mildews which cause late
blight of potatoes and buckeye rot of tomatoes.
The Zygomycetes from zygospores by the union of
the tips of two hyphae which often appear similar
and which may come from the same mycelium or
from different mycelia. Oospores and zygospores
are covered by a tough wall and can survive drying
for long periods.

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Life Cycle of a Zygomycete: Black Bread Mold (Rhizopus)
Reproduces Asexually and Sexually
Reproductive Structures of Zygomycete (Rhizopus)
Sporangia (asexual) and Zygospore (sexual)
Cultural characteristics
Some molds are loose and fluffy, others are
compact. Some look velvety on the upper surface,
some dry and powdery, and others wet or
gelatinous. Definite zones of growth in the thallus
distinguish some molds e.g., Aspergillus niger.

Pigments in the mycelium, red, purple, yellow, Gray,


black, brown.

Pigments of masses of asexual spores: green, blue


green, yellow, orange, pink, lavender, brown, gray,
black.
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Physiological Characteristics
Moisture requirements
Below 14 to 15 percent total moisture in flour or some dried fruits
will prevent or greatly delay mold growth.
Temperature requirements
Most molds would be considered mesophilic i.e., able to grow well
at ordinary temperatures. The optimal temperature for most molds
is around 25 to 30 C, some at 35 to 37 C or above, e.g., Aspergillus
spp.

Psychrotropic i.e., grow fairly well at temperature of refrigeration, and some


can grow slowly at temperatures below freezing. Growth has been reported
at as low as -5 to -10 C.

Thermophilic i.e., they have a high optimal temperatures.

Oxygen and pH requirements


Molds are aerobic. Most molds can grow over a wide range of
hydrogen-ion concentration (pH 2 to 8.5), majority are favored by
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and acid pH.


Food requirements
Molds in general can utilize many kinds of foods,
ranging from simple to complex. Molds possess a
variety of hydrolytic enzymes, some are grown
for their amylases, pectinases, proteinases, and
lipases.
Inhibitors
Compounds inhibitory to other organisms are
produced by some molds, such as penicillin from
Penicillium chrysogenum and clavacin from
Aspergillus clavatus. P. griseofulvin
griseofulvin
Cephalosporium cephalosporia
Certain chemical compounds are mycostatic, 18
The synthesis of toxins such as:
a.Ergot alkaloids- these are produced by
Claviceps
purpurea and cause an alpha adrenergic
blockade.
b. Psychotropic agents - these include
psilocybin,
psilocin and lysergic acid diethylamide
(LSD).
c.Aflatoxins - these are carcinogens
produced by
Aspergillus flavus when growing on grain. 19
Penicillium chrysogenum

Aspergillus clavatus 20
Classification and identification of molds
Kingdom: Myceteae, belong to the Eumycetes, or true fungi.
No roots, stems, or leaves and are devoid of chlorophyll.

Criteria are used for differentiation and


identification of molds:
1. Hyphae septate or nonseptate
2. Mycelium clear or dark (smoky)
3. Mycelium colored of colorless
4. Whether sexual spores are produced and the type:
zygospores, oospores or ascospores
5. Type of asexual spores: sporangiospores, conidia, or
arthrospores (oidia)
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6. Characteristic of the spore head
a.sporangia: size, color, shape and location
b.spore heads bearing conidia: single conidia, chains,
budding conidia, or masses; shape and arrangement of
sterigmata or phialides; gumming together of conidia.
7. Appearance of sporangiophores or conidiophores: simple
or branched.
8. Microscopic appearance of the asexual spores,
especially of conidia: shapes, size, color, smooth or
rough; one-, two-, or many celled
9. Presence of special structures (or spores): stolon,
rhizoids, foot cells, apophysis, chlamydospores, sclerotia.

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Molds of industrial importance
Mucor
M. racemosus, M. rouxii is used in the amylo process for the
saccharification of starch, and help ripen some cheeses (e.g., Gammelost)
and are used in making certai oriental foods.

Zygorrhynchus
These soil molds are similar to Mucor except that the zygospore suspensors
are markedly unequal in size.

Rhizopus
R. stolonifer (bread mold), involved in the spoilage of many foods: berries,
fruits, vegetables, bread.

Absidia
Similar to Rhizopus, except that sporangia are small and pearshaped.

Thamnidium
T. elegans is found on meat in chilling storage, causing whiskers on the
meat. 23
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Aspergillus
Many are involved in the spoilage of foods, some are
useful in the preparation of certain foods. A. glaucus
and
A. repens as an important species, often involved in
food
spoilage. A. niger, important in foods. A. flavus-oryzae,
oriental foods.
Penicillium
P. expansum, the blue green spored mold, caused soft
rots of fruits.
P. digitatum, with olive-or yellowish green conidia,
causing a soft rot of citrus fruits.
P. italicum (blue contact mol), blue green conidia,
rotting
citrus fruits.
P. camemberti, grayish conidia, ripening of Camembert25
Mucor
Zygospore

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Sporangiospores Sporangium

Columella

Sprorangiophore

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Absidia

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Sporangiospores
sporangium

columnella

sporangiophore
Absidia

node

Rhizoid in between
sporangiophores

stolon

Rhizopus
Rhizopus -bread30
Trichothecium
T. roseum, pink, grows on wood, paper,
fruits (apples and peaches) and
vegetables (cucumbers and
cantaloupes). The clusters of two-celled
conidia at the ends of short, erect
conidiophores.

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Geotrichum (Oospora or Oidium)

Genus, yeastlike fungi (some writers)


and molds (others). Species: white,
red, yellowish or orange. Growth
appearing first as a firm, feltlike mass
that later becomes soft and creamy.
G. candidum (Oospora lactis) or diary
mold, white to cream. The hyphae
are septate and in common species
are dichotomously branched. The
asexsual spores are arthrospores
(oidia), which may appear rectangular33
Oval arthrospores (aerial)

Cylindrical arthrospores (submerged)

Geotrichum
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Neurospora
(Monilia)

Under genus
Neurospora.
Neurospora
(Monilia) sitophila,
important in food,
red bread mold
because its pink,
loose textured
growth often
occurs on bread.35
Sporotrichum
Among the
saprophytic
species is S.
carnis, found
growing on
chilled meats,
where it
causes white
spot.
36
Botrytis

B. cinerea,
important in
food. It cause a
disease of
grapes but may
grow
saprophytically
on many food.
37
Cephalosporium
C. acremonium is a common
species. 38
Trichoderma
T. viride, common
species.
The mature mold plant is
bright green because the
balls of green conidia are
glued together, and tufts
of white hyphae (sterile)
stick up well above the 39
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Scopulariopsis

S. brevicaulis, common species.


This genus may be confused with Penicillium, for
both have brush like penicilli and chains of spores
cut off from the sterigmata, the conidia of
Scopulariopsis are never green. The spores are
distinctive in microscopic appearance and are not
green, commonly yellowish-brown; they are lemon
shaped, thick-walled, spiny, and pointed at one end,
with a thick ring at the opposite end. Colonies are
brownish and cottony.

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Pullularia

P. pullulans, common
species. Ovate,
hyaline conidia
(blastospores or
buds from
preexisting
cells) borne as
lateral buds on all
parts of the
mycelium.
Colonies are pale 43
Cladosporium

C. herbarum,
leading species.
Dark molds cause
black spot on a
number of foods,
on cellar walls.
Colonies of C.
herbarum are
restricted in
growth and are
thick, velvety, and 44
olive to gray-
Helminthospo
rium

Species this genus


are for the most
part plant
pathogens. Grow
saprophytically on
vegetable
materials. 45
Alternaria

A.tenuis and A.
brassicae,
common species.
Common
causes of the
spoilage of
foods. A. citri
(rotting citrus
fruits). The mass of
mycelium usually is
dirty 46
gray-green, hyphae
Stemphylium
(common genus)

The conidia are


dark and
multicellular,
fewer cross walls
than those of
Alternaria and
are rounded at
both ends.
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Fusarium
Often grow on foods. The species are
very difficult to identify, appearance
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of growth is variable.
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Endomyces
Yeastlike fungi, forming mycelium and
arthrospores. Some species rot fruits.

Monascus
Colonies of M. purpureus are thin and
spreading and reddish or purple. Found
on diary products and on Chinese red rice
(ang-khak).

Sclerotinia
Cause rots of vegetables and fruits,
present in the conidial stage. The lemon-
shaped conidia are in chains, with plug
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ECONOMIC IMPORTANCE OF FUNGI
25-50% of harvested fruits and vegetables are damaged by
fungi.
Fungal infections of plants are commonly called rots, rusts,
blights, wilts, and smuts.
Phytophthora infestans: Caused great potato famine in mid-1800s.
Over 1 million people died from starvation in Ireland. Many
immigrated to the U.S.

Beneficial fungi:
Candida oleophila: Prevents fungal growth on harvested fruits.
Saccharomyces cerevisiae: Used to make bread and wine.
Genetically engineered yeast strains are used to make proteins
(Hepatitis B vaccine).
Taxomyces: Produces anticancer drug taxol.
Trichoderma: Produces cellulase. Used to make fruit juice.
Yeast
and
Yeastslike
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Yeasts maybe useful or harmful in
foods.

Yeast fermentations are involved


in the
manufacture of foods such as
bread, beer, wines,
vinegar, and surfaced ripened
cheese, and yeasts
are grown for enzymes and for
food. 53
General Characteristics
of Yeast
Morphological
Caracteristics
They are determined by
microscopic examination.

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Yeast - Major Characteristics

Unicellular Fungi
Eukaryotic
Facultative anaerobes
Capable of forming
colonies on solid culture
media (see pictures on the
right).
Occur worldwide
Over 1,500 species
described
www.furryhelpers.com/SJSU/Micro20/Lab5
-Yeast.ppt
Yeast - Reproduction
They reproduce either asexually (most common) or sexually.
Asexual reproduction is through budding or binary fission.
Sexual reproduction (if any) results in the formation of the appropriate spore
structure.

Fission
Budding

Saccharomyces cerevisiae Spores


Schizosaccharomyces octosporus
Yeast Significance
Food Industry
Fermentation of bread, beer, and wine. E.g. Saccharomyces cerevisiae (also
called bakers yeast or sugar yeast) used in baking and fermenting of alcoholic
beverages.

Medical
E.g. Candida albicans - common in the human mouth, but can become
pathogenic and cause Candidiasis (oral and/or genital infection).

Biofuel Industry
Production of ethanol for car fuel.
Yeast - Taxonomy
Yeasts are identified primarily by their biochemical
properties.
They are found only in 3 groups of fungi:
Ascomycotina
Examples: Saccharomyces cerevisiae
and Schizosaccharomyces octosporus
Deuteromycotina
Examples: Trigonopsis, Rhodotorula,
and Candida.
Basidiomycotina
The three Deuteromycotina
Examples: Trigonopsis, Rhodotorula, and Candida.

Trigonopsis :
Triangular cell morphology.
Budding yeast with buds arising at the apices.
Rhodotorula :
Orange/red pigmented colony morphology.
Budding yeast.
Candida :
Pseudohyphae formation this is formed by repeated budding process
where the buds do not separate from the parent cell.
This genus is an opportunistic pathogen and can cause a variety of
human infections.
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Reproduction
Most yeasts reproduce
asexually by multilateral
or polar budding, a
process in which some of
the protoplasm bulges
out the cell wall; the
bulge grows in size and
finally walls off as a new 63
64
Sexual reproductions of true yeasts (Ascomycotina)
results in the production of ascospores, the yeast cell
serving as the ascus. The usual number of spores
per ascus and the appearance of the ascospores are
characteristic of the kind of yeast. The ascospores
may differ in colour, in smoothness or roughness of
their walls, and in their shape (round, oval, reniform,
bean or sickle-shaped, saturn or hat-shaped,
hemispherical, angular, fusiform or needle-shaped).

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sexual cycle
Haploids can mate (or fuse)
to form diploid cells.

Two step Process:


1. Conjugation or
cytoplasmic fusion
2. Karyogamy or nuclear
fusion

httpwww.iupui.edu~biomg817contentYeast_
Biology.pdf
66
False yeasts, produce no ascospores
or other sexual spores, belong to the
Fungi Imperfecti. Cell of some yeasts
become chlamydospores by formation
of a thick wall about the cell, ex.
Candida, Rhodotorula and
Cryptococcus.

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Cultural Characteristics
Yeasts are oxidative, fermentative, or
both. The oxidative yeasts may grow as
a film, pellicle, or scum on the surface
of a liquid and then are termed film
yeasts. Fermentative yeasts usually
grow throughout the liquid and produce
carbon dioxide.
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Physiological Characteristics
Most common yeasts grow best with a plentiful
supply of available moisture. Yeasts classified as
ordinary if they do not grow in high
concentrations of solutes (such as sugar or salt),
i.e., in a low aw, and as osmophilic if they do.

Range from 0.88 to 0.94. Specific examples of


minimal aw are 0.94 for a beer yeast, 0.90 for a
yeast from condensed milk and 0.905 for a
bakers yeast. By contrast, aw as low as 0.62 to
0.65 in syrups, some osmophilic yeasts are
stopped at about 0.78 in both salt brine and
sugar syrup. 69
The range of temperature for growth of most yeasts
is, in general, similar to that for molds, around 25 to
30 C and maximum about 35 to 47 C.
The growth of most yeasts is favored by an acid
reaction in the vicinity of pH 4 to 4.5, not grow well
in alkaline medium unless adapted to it.
Yeasts grow best under aerobic conditions, but the
fermentative types can grow anaerobically, although
slowly.
In general, sugars are the best source of energy for
yeasts. The yeasts also aid in the production of
flavors or bouquet in wines.
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Classification and Identification of Yeasts
The true yeasts are in the subdivision Ascomycotina.
The false yeasts are in the subdivision Fungi Imperfecti or
Deuteromycotina.
Several examples, with equivalent names
Diploid Haploid
Perfect (sexual) stage Imperfect (asexual) stage
1. Kluyveromyces marxianus 1. Candida kefyr (syn.
(syn. K.fragillis & S.fragillis) C.pseudotropicalis)

2. K.marxianus var.lactis (syn. 2. Candida sphaerica (syn.


S.lactis) Torulopsis candida)

3. Debaryomyces hansenii 3. Candida famata (syn.


T.candida)

4. Pichia burtonii 4. Candida variabilis (syn. 71


C.fibrae)
The principal bases for the
identification and classification of
genera of yeasts:
1.Whether ascospores are formed.
2.If they are spore-forming:
a.The method of production of ascospores:
1.Produced without conjugation of yeast cells
(partenogenetically)
Spore formation by:
a.conjugation of ascospores.
b.conjugation of small daughter cells.
2.Produced after isogamic conjugation
(conjugating cells appear
similar).
3.Produced by heterogamic conjugation
(conjugating cells differ 72
in appearance).
4.Method of asexual reproduction:
a.Budding.
b.Fission.
c.Combined budding and fission.
d.Arthrospores (oidia).
5.Production of a mycelium, pseudomycelium,
or no mycelium.
6.Growth as a film over surface of a liquid (film
yeasts) or growth
throughout medium.
7.Colour of macroscopic growth.
8.Physiological characteristics:
a.Nitrogen and carbon sources.
b.Vitamin requirements.
c.Oxidative or fermentative: film yeasts are
oxidative; other
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yeasts may be fermentative or fermentative
Yeasts of Industrial Importance
Most yeasts used industrially are in the
genus Saccharomyces. The term wild
yeast is applied to any yeast other than the
one being used or encouraged. Most of the
troublesome wild yeasts are
asporogenous, or false, yeasts.

Genus Schizosaccharomyces
A common species is S.pombe. They have
been found in tropical fruits, molasses,
soil, honey, and elsewhere. 74
Genus Saccharomyces
The leading species, S.cerevisiae, is
employed in
many food industries, with special
strains used for the leavening of
bread, as top yeasts for ale, for wines,
and for production of alcohol, glycerol,
and invertase. Top yeasts are very
active fermenters and grow rapidly at
20 C. The clumping of the cells and
the rapid evolution of CO2 sweep the
cells to the surface, hence the term 75
76
S. cerevisiae var. ellipsoideus is a high-
alcohol-
yielding variety used to produce
industrial alcohol,
wine, and distilled liquors. S. uvarum, a
bottom
yeast, is used in making beer. S. fragilis
and S. lactis, because of their ability to
ferment lactose, important in milk or
milk products.
Genus Zygosaccharomyces
These yeasts are notable for their ability
to grow in high concentrations of sugar 77
and are involved in the spoilage of
Genus Pichia
P. membranaefaciens grows a pellicle on
beers or wines.

Genus Debaryomyces
D. kloeckeri grows on cheese and sugarcane.

Genus Hanseniaspora
These lemon-shaped (apiculate) yeasts grow
in fruit juices. Nadsonia yeasts are large and
lemon-shaped.
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False Yeasts (Fungi
Imperfecti)
Genus Torulopsis
T. sphaerica ferments lactose and may
spoil milk products. Other species can
spoil sweetened condensed milk, fruits-
juice concentrates, and acid foods.

Genus Candida
Many form films and can spoil foods high
in acid and salt. C. utilis is grown for food
and feed. C. krusei has been grown with
dairy starter cultures to maintain the 79
Genus Brettanomyces
They are involved in the late fermentation of Belgian
lambic beer and English beers. They also are found in
French wines. B. bruxellansis and B. lambicus are typical
species.
Genus Kloeckera
K. apiculata is common on fruits and flowers and in the
soil.
Genus Trichosporon
They grow best at low temperatures and are found in
breweries and on chilled beef. T. pullulans is a common
species.
Genus Rhodotorula
These red, pink, or yellow yeasts may cause
discolourations on foods, e.g., coloured spots on meats
80
or pink areas in sauerkraut.
Groups of Yeasts
Film yeasts, in the genera Pichia, Hansenula,
Debaryomyces, Candida, and Trichosporon, grow on the
surface of acid
products such as sauerkraut and pickles, oxidize the
organic acids, and enable less acid tolerant organisms to
continue the spoilage.
Saccharomycodes, Hanseniaspora, Nadsonia, and
Kloeckera, are considered objectionable in wine
fermentations because they give off-flavors, low yields of
alcohol, and high volatile acid.
The most salt tolerant of the film yeasts are species of
Debaryomyces, which grow on curing brines and on
meats and cucumbers in them. S. rouxii can grow as a
film on brine, considered of great importance in the
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productions of alcohol and flavor.
Bacteria

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Bacterias life
Bacteria are all around us. Given good growing
conditions,
a bacterium grows slightly in size or length,
a new cell wall grows through the center forming two
daughter cells, each with the same genetic material
as the parent cell.
If the environment is optimum, the two daughter
cells may divide into four in 20 minutes. 1, 2, 4, 8, 16,
32, 64...
Microbial Growth
refers to the growth of
a population (or an
increase in the
number of cells),
not to an increase in
the size of the
individual cell.
Cell division leads to
the growth of cells in
the population.
Two Types of Asexual Reproduction in
Microbes:
1.)Binary Fission
2.) Budding
1.)Binary Fission
Bacterial reproduction occurs through fission, a primitive
form of cell division that does not employ a spindle fiber
apparatus.
A spindle fiber apparatus made of protein filaments is
responsible for moving the chromosomes around during
cell division (mitosis & meiosis) in most eukaryotic cells.
Bacteria do not have these structures.
The bacterial cell doubles in size and replicates its
chromosome.
Following DNA replication, the two chromosomes attach
to separate sites on the plasma membrane, and the cell
wall is laid down between them, producing two daughter
cells.
2.) Budding

A few bacteria and some eukaryotes (including


yeasts) may also replicate by budding,
forming a bubble-like growth that enlarges and
separates from the parent cell.
Phases of Growth
A microbial lab culture
typically passes through
4 distinct, sequential
phases of growth that
form the standard
bacterial growth curve
1.Lag Phase
2.Log Phase
3.Stationary Phase
4.Death Phase
Phases of Growth
LAG PHASE: Growth is slow at first, while the
"bugs" acclimate to the food and nutrients in
their new habitat.
LOG PHASE: Once the metabolic machinery is
running, they start multiplying exponentially,
doubling in number every few minutes.
STATIONARY PHASE: As more and more bugs
are competing for dwindling food and nutrients,
booming growth stops and the number of
bacteria stabilizes.
DEATH PHASE: Toxic waste products build up,
food is depleted and the bugs begin to die.
At 11:00, we place a single bacterium in a
bottle. It's so small you'd need a microscope to
see it.
In one minute it grows to twice its original size
and divides in half, reproducing itself, so at
11:01 there are two bacteria in the bottle.
At the end of five minutes, there are 32 bacteria where there used
to be just one ... but even all together they're still so small they can't
be seen without a microscope.
The bacteria keep doubling their numbers
every minute,
until 12:00, when the bottle fills up.
When the bottle has half full of bacteria?
That's right! Since the bottle filled up at 12:00, it must have been half
full just a minute before.

At 11:45 we could just barely see the bacteria, and at 11:30 we still
needed a microscope!
Classification of Bacteria

Until recently classification has done on the basis of such


traits as:
shape
~bacilli: rod-shaped
~cocci: spherical
~spirilla: curved walls
ability to form spores
method of energy production (glycolysis for anaerobes,
cellular
respiration for aerobes
nutritional requirements
reaction to the Gram stain.
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The Gram stain is named after the 19th century Danish
bacteriologist who developed it.

The bacterial cells are first stained with a purple dye called
crystal violet.
Then the preparation is treated with alcohol or acetone.
This washes the stain out of Gram-negative cells.
To see them now requires the use of a counterstain of a
different color (e.g., the pink of safranin).
Bacteria that are not decolorized by the alcohol/acetone
wash are Gram-positive.

Although the Gram stain might seem an arbitrary criterion to


use in bacterial taxonomy, it does, in fact, distinguish
between two fundamentally different kinds of bacterial cell
walls and reflects a natural division among the bacteria.
97
Gram-positive bacteria are encased in a plasma
membrane covered with a thick wall of
peptidoglycan.

Gram-negative bacteria are encased in a triple-


layer. The outermost layer contains
lipopolysaccharide (LPS).
98
Morphological Characteristics
Important in
Food Bacteriology
Microscopic examination to ascertain the shape,
size,
aggregation, structure, and staining reactions of
the bacteria present.

Encapsulation
The presence of capsules or slime may account
for sliminess or ropiness of a food. To increase
the resistance of bacteria to adverse conditions,
such as heat or chemicals. Most capsules are
polysaccharides of dextrin, dextran, or levan.

Formation of Endospores 99

Bacteria of genera Bacillus (aerobic and some


Endospores are formed at an intracellular site,
are very
refractile, and are resistant to heat, ultraviolet
light, and
desiccation.

The complete cycle of vegetative cell through


sporulation on the free spore, possible extended
dormancy, and then germination of the spore
and outgrowth back to the vegetative cell is
extremely complex.

Sporulation usually appears in the late


logarithmic phase of growth, possibly because of
nutrient depletion or product accumulation.
During this transition of vegetative cell to spore,
the spore becomes refractile, there is a massive100
uptake of Ca2+ ions, and synthesis of dipicolinic
Germination is inhibited by sorbic acid at an
acid pH, by
some divalent cations, by starch, and by oleic
and linoleic
acids. The spores fail to germinate, because
unfavorable conditions such as inhibitors in
the environment or lack of essential
nutrients, e.g., amino acids. Delayed
germination of spores from a few days to
many months has been reported, e.g.,
dormancy from a few days to 3 or 4 months
with spores of Bacillus megaterium and for 15
days to 72 months with spores of Clostridium
botulinum.

Dormancy of spores has been defined as


delayed germination under conditions 101
apparently favorable for it.
Cultural Characteristics Important
in Food Bacteriology
Pigmented bacteria cause discolorations on
the surfaces of foods; films may cover the
surfaces of liquids; growth may make surface
slimy; or growth throughout the liquids may
results in undesirable cloudiness or sediment.

Physiological Characteristics
Important in Food Bacteriology
Hydrolysis of complex carbohydrates to
simple ones; hydrolysis of proteins to
polypeptides, amino acids, and ammonia or
amines; and hydrolysis of fats to glycerol and
fatty acids. O-R reactions, which are utilized
102
by the bacteria to obtain energy from foods,
Genera of Bactera Important in
Food
Bacteriology
Genus Acetobacter
Oxidize ethyl alcohol to acetic acid, found on
fruits, vegetables,
souring fruits, and alcoholic beverages. They
are a definite
spoilage problem in alcoholic beverages.
Genus Aeromonas
These are gram-negative rods with an
optimum tempertaure for
growth of 22 to 28 C. Isolated from aquatic
environments.
A.hydrophila can be a human pathogen; it is
also pathogenic to 103
Genus Bacillus
The thermophilic flat sour bacteria that spoil
canned vegetables can produce considerable
amounts of lactic acid from sugar, and sch a
culture, e.g., B. coagulans, employed for the
manufacture of lactic acid. The soil is a
important source of Bacillus species.
Several strains, as recognized by American Type
Culture Collection (ATCC) number, are important
as test organisms in sterility testing.

B. pumilus (ATCC 27142) is recommended for


determining the suitability of gamma radiation
sterilization.
B. stearothermophilus (ATCC 7953) is
recommended for testing procedures involving 104
steam sterilization.
Genus Brochotrix
These gram-positive rods. The organisms will
not survive heating at 63 C for 5 min. They
can spoil a wide variety of meats and meat
products when they are stored aerobically or
vacuum packed and held refrigerated. B.
thermosphacta is the only species listed.

Genus Clostridium
C. thermosaccharolyticum is saccharolytic
obligate thermophile; this organisms causes
gaseous spoilage of canned vegetables.
Putrefaction of foods often is caused by
mesophilic, proteolytic species, such as
C.lentoputrescens and C. putrefaciens. The
violent disruption of the curd in milk by C.
perfringens or similar species results in a 105

stormy fermentation of milk, and the


Genus Corynebacterium
The diphtheria organism, C. diptheriae,
transported by foods.
C. bovis, with the slender, barred, or clubbed
rods
characteristic of the genus, is common on the
cow`s udder,
found in aseptically drawn milk, a cause of
bovine mastitis.

Genus Desulfotomaculum
D. nigrificans, is responsible for sulfide stinker
spoilage in canned foods. Sulfur sompounds can
serve as the terminal electron acceptor in
respiration and thereby be reduced to hydrogen
sulfide.

Genus Erwinia
This genus are plant pathogens that cause 106
necrosis, galls, wilts, or soft rots in plants and
Genus Escherichia
Found in feces, a predominant
gram-negative rod isolated from the
intestinal tract of warm-blooded
animals and widely distributed in
nature. Some of which can be
pathogenic to humans.

Family Lactobacillceae
Genus Lactobacillus
Growing well at higher
temperatures (37 C or above), L.
bulgaricus, L. helveticus, L. lactis, L. 107
Charcteristics that make the
lactobacilli important in foods:
1.Their ability to ferment sugars with
the
production of considerable
amounts of lactic acid.
2.Production of gas and other volatile
products by
heteroferentative species,
sometimes damage to
the quality of food, L. fermentum
growing in
Swiss cheese or L. hilgardii or L.
trichodes in 108

wines.
Genus Leuconostoc. Important in foods:
1.Production of diacetyl and flovoring
products.
2.Tolerance of salt concentrations.
3.Ability to initiate fermentation in vegetable
products more
rapidly than other lactics.
4.Tolerance of high sugar concentrations (up
to 55 to 60% for
L. mesenteroides).
5.Production of considerable amount carbon
dioxide gas from
sugars and leavening some breads.
6. Heavy slime production in media
containing sucrose.
Genus Salmonella
109
Grow in foods and cause food infections.
Genus Pseudomonas
Pseudomonas can cause food spoilage.
Important in foods:
1.Ability to utilize of noncarbohydrate carbon
compounds for
energy and inability to use most
carbohydrates.
2.Ability to produce a variety of products that
affect flavor
deleteriously.
3.Ability to use simple nitrogenous foods.
4.Ability to synthesize their own growth factors
or vitamins.
5. Proteolytic and lipolytic activity.
6.Aerobic.
7.To grow well at low temperatures.
110
8.Pigment production, e.g., greenish
Genus Staphylococcus
The most important species,
aureus, give yellow to orange
growth. Causes food poisoning.

Genus Streptococcus
The streptococci important
in foods are divided into
four groups: the pyogenic,
viridans, lactics,
enterococcus. 111
Groups of Bacteria Important in Food
Bacteriology
1.Lactic acid-forming bacteria, or lactics
(Leuconostoc,
Lactobacillus, Streptococcus, Pediococcus).
2.Butyric acid-forming bacteria, or butyrics
(genus
Clostridium).
3.Propionic acid-forming bacteria, or propionic
(genus
Propionibacterium).
4.Proteolytic bacteria (B. cereus, Clostridium,
Pseudomonas,
Proteus).
5.Pigmented bacteria (Flavobacterium, yellow112
to orange;