M O LEDX A MA

I NN DO N YO FE A S T
ATI

CULTURES
Experiments performed

Isolation of yeast and molds from

three food samples

Wet mount method

Slide culture method

Yeast inoculation into sugar broth

Ascospore staining
 2 3
1 They are highly
They are utilized in
Both of them considered one various
belong to the of the main industries for
kingdom fungi, reasons for their beneficial
though they spoilage and properties and
have different decomposition wide range of
characters and of organic products that
uses. matter. can be
synthesized from
them.
 Mold vs YEAST

More than 100,000 About 1,500

different species different species
present present

Multicellular Mostly
unicellular

Comprises of Mostly consist of

hyphal filaments pseudohyphae
and spores

Exhibits both Could exhibit

sexual or asexual asexual
reproduction reproduction and
sexual reproduction
Hyphae - branches
or filaments
which
collectively
known as mycelium

Pseudohyphae –
product of an
incomplete
budding where the
cells elongate
but remain
attached after
division.

Spores - organs
of sexual and
asexual
reproduction
Hyphae - branches
or filaments
which
collectively
known as mycelium

NON-SEPTATE
SEPTATE
( C OPpEsreNoudO
dohyphae –
CYTIC)
uct of an
incomplete
budding where the
cells elongate
but remain
attached after
division.

Spores - organs
Branching: lateral or apical
of sexual and
asexual
reproduction
 Hyphae - branches
or filaments
which
collectively
known as mycelium

NON-SEPTATE
SEPTATE
(COENOCYTIC)

Spores - organs
Branching: lateral or apical
of sexual and
asexual
reproduction
Spores - organs
of sexual and
asexual
reproduction
 Spores - organs
of sexual and
asexual
reproduction

conidia sporangiospores arthospores ascospores

 Sporangium

Spores Stolon

Rhizoids
Columella

Sporangiophore

Other structures
 Food
safety
SIGNIFICANCE OF MOLD AND YEAST IN FOOD MICROBIOLOGY

Food
production
 Food
safety
SIGNIFICANCE OF MOLD AND YEAST IN FOOD MICROBIOLOGY

Food
production
Food Inflict hazard to
3
safety
humans due to
toxin production,
allergic reaction,
and infection

Thrive in plants,
Cause 4 which produce
1 decomposition
deterioration
and
of
products with high
sugar content, low
foods water activity,
and low pH

Grow and invade Breed in food

2 crops and
generally, any 5 processing
environment when
type of food under poor sanitation
favorable practices are
condition present
 Food Inflict hazard to
3
safety
humans due to
toxin production,
allergic reaction,
and infection

Thrive in plants,
Cause 4 which produce
1 decomposition
deterioration
and
of
products with high
sugar content, low
foods water activity,
and low pH

Food
2
Grow and invade
crops and
5
Breed in food
processing

production
generally, any environment when
type of food under poor sanitation
favorable practices are
condition present
 Food
safety

Food
production
 Main component in the
1 production of alcoholic
beverages, bread and fermented
products

Provide a large variety of

2 industrial products such as
antibiotics, enzymes,
vitamins,
and organic acids

Food
production
WIDELY USED MOLD AND YEAST
IN FOOD INDUSTRY
YEAST
S. cerevisae – also known as brewer’s yeast
S. boulardii – used as probiotic
D. hansenii – reduce mycotoxin contamination in
foods

MOLDS
A. oryzae – used in production of koji, miso,
soybean paste and soy sauce
R. oligosporus – used in production of bean cake
or bean cheese (sufu) and tempeh
P. roqueforti – used in cheese making
t
Objectives
O F T H E E X E R C I S E

1 Isolate yeast and mold colonies

in a given food sample.

Understand the characteristics of

2 yeast and mold cultures based on
the analysis of its morphology
and development and lastly,

Perform the procedures such as

3 microscopy, serial dilution,
slide culture staining and colony
counting
 Mold cultures

M u c o r R h i z o p u s

F u s a r i u m A l t e r n a r i a

P e n i c i l l i u m T r i c h o d e r m a
 Slide culture Wet mount method
method
Useful for giving clear
images of fresh specimens
Focuses on the under the microscope e.g.
microscopic spores of fungi and ferns,
observation of and pollen grains may be
sporulation of a best observed using this
colony technique

Note: Motility needs to be

Allows keen distinguished from Brownian
observation with motion which is due to
minimal amount of molecular bombardment.
disturbance and Brownian motion occurs in
damage of its all microscopic bodies
structure suspended in water and
appears as a random
movement. Motility will be
Suggested method for in the form of cork-screw
characterizing spiraling, movement in a
morphology of fungal given direction, or
 mucor
• White to grayish brown mycelia
• Non-septated
• Sporangiophores, sporangia, and
spores are present
• Apophysis, rhizoid and stolon
are absent
• Sporangiospores are round or
slightly elongated
 Fusarium
• White to pink surface mycelia
• Septated
• Single sterigmata are present in
numerous spores
• Asexual spores, conidia, are
present
microconidia - cylindrical abundant
non-septated
macroconidia - boat-shaped and
septated
 penicillium
• Blue green and velvety mycelia
• Septated
• Simple or branched conidiophores
are visually present
• Round and unbranched conidia are
present
• Flask-shaped phialides carried by
the metulae are present
 rhizopus
• Yellowish brown and cotton-like
mycelia
• Non-septated
• Sporangiophores, rhizoids (root-
like hyphae), and sporangia are
visualized
• Apophysis is absent and columellae
are hemispherical.
• Sporangiospores are ovoid-shaped
and smooth in texture
 alternaria
• Grayish and woolly mycelia
• Septated
• Brown condiophores that are zigzag
in appearance are present
• Simple or branched conidia, which
are ovoid-shaped are present
 trichoderma
• Blue-green or yellow-green patches
of mycelia
• Septated
• Branched conidiophores are present
in pyramid arrangement
• Flask-shaped phialides are
arranged in clusters
• Ellipsoidal-shaped conidia are
present
 Medium used
Potato Dextrose Agar (PDA)
PDA can be used for growing clinically
significant yeast and
molds. The nutritionally rich base (potato
infusion) encourages mold sporulation and
pigment production.

Able to work with acid or antibiotics to

inhibit bacterial growth (Chloramphenicol)
 Medium used
Nutrient Sugar Broths (Glucose, Sucrose and
Lactose)
Peptone – used as an organic nitrogen source
for microorganisms

Beef Extract – recommended for use in

bacteriological examination of water, milk,
and other materials
- relied upon for biochemical studies,
particularly fermentation reactions because
of its independence from fermentable
substances

Sugars – source of carbon and energy

 Medium used Ascospore
Lactophenol staining
Specialized for fungal staining
and wet mounting molds Malachite green is
used for staining
to identify the
presence of
Lactic acid - preserves the
ascospores in yeast
fungal structure
species.

Phenol – used as disinfectant

Malachite green dye
is water-
soluble and does
Cotton blue – stains the
not adhere well to
structure particularly chitin
the cell wall
and cellulose
making it easily
rinsed.

In most cases,
safranin serves as
 yeast isolates
G L U C O S E L A C T O S E S U C R O S E

R h o d o t o r u l a - , tubid r o m y c e- s, slightly turbid

D e b- ,atubid

- , tubid - , clear - , clear

S . p o m b e - , tubid - , Pclear
i c h a - , clear

+ , tubid - , clear + , turbid

S . c e r e v i s i a e - , tubid - , slightly turbid + , turbid

+ / - o f p e l l i c l e
f o r m a t i o n
Pellicle formation
Pellicle is the ropy-like or web-like layer on
1 top of the nutrient medium. It has uneven
appearance and can contain bubbles.

Biofilm - community of microorganisms that are

2 connected to one another through the use of an
extracellular matrix (material outside of the
cell)

The purpose of this formation is so the

3 cells can undergo oxidative metabolism in
order to access the little oxygen present
in the atmosphere of the fermenter.
Yeast characterization
1 Since yeast isolates usually appear as circular
in shape, it is quite hard to characterize among
species.

Assimilation test allows the identification of

2 yeast isolates that are slow metabolizers. Plus,
it determines the capability of the organism to
use a particular substrate.

Other methods of characterization include:

3 Auxanographic method, Rapid Screening Test,
Chromogenic media, Automated method,
PCR-based assays
M O LEDX A MA
I NN DO N YO FE A S T
ATI

CULTURES