Definition

• Mykos (mushroom)
• Study of fungi which include molds
and yeasts
 General Characteristics
• Eukaryotic organisms
• Exist as parasites or saprophytes
(feed on organic substance or decay)

• Reproduction
• Asexual
• Sexual
 Nutritional Requirements
• Chemoheterotrophs
– Obtain their nutrients by absorbing
chemicals found in the environment
(organic compounds)

– Fungi secrete enzymes that degrade

organic substrates into soluble
nutrients
 Answer the following:
• Bacterial cultures are incubated at _________
• Fungal cultures, on the other hand, are
incubated at room temperature

• Bacteria culture medium have pH _____ while

fungal culture medium have acidic pH
 Answer the following:
• Bacteria, if placed on a hypotonic solution will
__________ while fungi are resistant to
osmotic pressure.

• Bacteria can be (depending on oxygen

requirement) _________ or ____________
– Yeast are facultative anaerobes
– Molds are aerobic
 NOTE:

• Bacterial growth greatly

depends on the availability of
nitrogen and moisture
 Nutritional Requirements
Fungi Differ From Bacteria

• Fungi grow best at room temperature

– Bacteria prefer to grow at 37 degree Celsius

• Fungi grow best with a pH of 5

– Bacteria grow best at pH 6.5-7.0
 Nutritional Requirements
Fungi Differ From Bacteria

• Almost all molds are aerobic; most yeasts

are facultative anaerobes (does not need
oxygen but can survive in its presence)
– Bacteria can be anaerobe or aerobe

• Most fungi are more resistant to osmotic

pressure than bacteria
 Nutritional Requirements
Fungi Differ From Bacteria
• Fungi can grow on substances with a very low
moisture content while bacteria cant.
• Fungi require less nitrogen than bacteria for
growth
• Fungi are often capable of metabolizing
complex carbohydrates that most bacteria
cannot use for nutrients
– Ex. Lignin (a component of wood)
 Harmful Effects
• Cause human disease in 3 general ways
– (1) Fungous allergies

– (2) Mycotoxicoses – disease cause by the toxin of a

fungus
• Aflatoxin which can cause hepatocellular carcinoma
– (3) Mycoses – infection of human body caused by
fungi
 Beneficial Effects
Medical and Economic Importance

• Yeasts (Saccharomyces) in preparation of

bread
• Preparation of vaccines (Hepatitis B vaccine)
• Sources of drugs
– Penicillin
– Cephalosporin
Bread
 Preparation of Hepatitis B
(Saccharomyces cerevisiae)
 Beneficial Effects
Medical and Economic Importance

• Higher fungi, mostly basidiomycetes

may be eaten directly as mushrooms

• Saprophytic fungi in soil produce

degradative enzymes essential for the
biologic recycling of organic matter
 Morphology

• Yeast – unicellular, grows best at

37 degree Celsius

• Molds – multicellular; grows best at

room temperature
 Dimorphism
• The ability of some fungi to grow in 2
forms under different environmental
conditions
• Grow as
– Yeast form at 37 C
– Mold form at room temperature
 Yeast
• Unicellular growth of
fungi
• Spherical or ellipsoidal
(3 – 15 micra)

• Most reproduce by
budding (forming
blastoconidia); few by
binary fission
 Yeast
• Most reproduce by
budding (forming
blastoconidia); few by
binary fission
– Buds fail to detach
producing elongated yeast
cells (pseudohyphae)
Pseudohyphae
 Hyphae versus pseudohyphae
• Hyphae have no
constrictions

• Hyphae is seen in
molds

• Pseudohhyphae is
demonstrated in
yeast form
 Yeast

• Colonies are pasty,

opaque, 0.3 – 5 mm in
diameter
• Most are cream-colored;
few species produce
pigments
 Molds
• Produce multicellular,
filamentous, irregular & dry
colonies

• Consist of branching
cylindrical tubules with
diameter from 2–10 micra,
called hyphae
 Molds

• Hyphae grow to form a filamentous mass

of intertwining strands called a
mycelium
 Mold growth
• Long filaments growing at apex branching. (sing.
Hypha): Mold
• Apical growth
–Hyphae branch to form mycelium
»Mycelium (plural mycelia)

www.myhome.cortes.com
MYCELIUM
www.rhodes.edu/biology/hill/hill/ResearchBackground.html
 Hyphae: 2 portions

• Vegetative portion or thallus,

– grows in or on a substrate and absorbs
water and nutrients
• Reproductive or aerial portion,
– project above the surface of the agar
medium and contains the reproductive
structures such as spores
 Hyphae

• Septate if with cross-

walls which divide the
hyphae into uninucleate
cell-like units
 Hyphae
• Nonseptate or
coenocytic if cross-walls
are absent and appear as
long continuous cells with
many nuclei
– Zygomycetes: Mucor,
Rhizopus, Absidia
Zygomycetes (Coenocytic hyphae)
 Hyphae

• Hyaline if fungal
structures are
colorless

• Highly refractile
 Hyphae
• Dematiaceous if fungi
produce melanin-like
pigments are dark-colored
• The term dematiaceous applies
to fungi that are dark in color.
• Many fungi causing human or
animal infections belonging to
this group are pale brown, dark
brown or black
 Hyphae
• Dematiaceous if fungi
produce melanin-like
pigments are dark-colored
• Agents causing
Chromoblastomycosis
– Phialophora
– Exophiala
– Curvularia
– Alternaria

– Brown CAPE
 Subcellular Structures
Capsule

• Composed predominantly of
polysaccharide

• Capsule of Cryptococcus neoformans has

antiphagocytic properties and is
associated with virulence
– Fungal meningitis among
immunocompromised patients
India ink staining
 India Ink Wet Mount

• Used to identify the capsule of Cryptococcus

neoformans
• CSF is directly examined by adding one drop
of India ink

• Capsule appear as a clear halo against a dark

background (negative staining)
White capsule against a black
background
 Cell Wall
• Comprise 15-30 % of the dry weight
of fungus
• Generally thicker in yeasts than in
molds
• Appears highly refractile under light
microscope
 Matching type
• 1. Bacteria • A. Cellulose
• 2. Plants • B. Peptidoglycan
• 3. Fungi • C. No cell wall
• 4. Humans • D. Chitin
Bacterial cell wall
Yeast cell wall
 Cell Wall
• 80% or more is carbohydrate
– Major component is chitin (N-acetylglucosamine)
– Varying amounts of glucan, cellulose and mannan

• The remaining components are protein and

glycoprotein
 Cell Wall
• Fungal cell wall is poorly stained with
routine H and E
• Fungal cell wall stains
– Periodic acid Schiff
– Methenamine silver stain
– Calcofluor white is a fluorescent stain
– Gram stain is useful for Candida
Gram stain - Candida
Methenamine silver stain - black
Periodic acid schiff - red
 Fluorescence Test

• Calcofluor white stain

–Calcofluor binds to chitin in cell
wall giving a brilliant
fluorescence
 Cell Membrane
• Bilayered membrane composed of
several phospholipids
• Contain sterols which are essential for
the viability of fungi
– Principal fungal sterols are ergosterol and
zymosterol
Human cell membrane
 Cytoplasmic Contents
• Nuclei, mitochondria, ER, storage vacuoles
containing hydrolytic enzymes, ions,
metabolites such as amino acids
Laboratory Identification of
Fungi
 Laboratory Methods

• Microscopic Methods
• Cultivation
• Biochemical tests
• Serologic Test
 Microscopic Methods
• Wet Mounts
–KOH
–Lactophenol cotton blue (LPCB)
–India ink
• Fluorescence Test
• Staining
10% Potassium hydroxide
 KOH (10%) Wet Mount
• Direct preparation from clinical specimen

• OH acts as clearing agent

– Dissolves keratin
– Eliminates debris

• One drop of KOH is added to specimen on a slide

– Allow specimen to clear for 20 minutes or warm slide
gently
• Disadvantage: poor contrast
Lactophenol Cotton Blue
 LPCB Wet Mount

• Lactic acid preserve the fungal structures

• Phenol is a killing agent
• Cotton blue color fungal structure

• Add one drop of LPCB to specimen on a

slide
• Advantage: impart a blue color to cell wall
 Fluorescence Test

• Wood’s lamp (UV light)

– Infected hair and skin will
fluoresce when examined in
the dark
 Cultivation
Requirements
• Culture media must include sources
of
– Nitrogen, Nitrate, Amino acids, Carbon,
Vitamins and minerals
– Antimicrobial supplements
• Cyclohexamide: inhibit contaminating
saprophytic fungi
• Chloramphenicol: inhibit bacterial growth
 Culture Media

• Primary isolation media

• Differential test media
 Primary Isolation Media
• Sabouraud dextrose agar (SDA)
– for primary isolation of saprophytic and
pathogenic fungi

–SDA-CC for recovery of pathogenic

fungi
• bacteria and saprophytic fungi are
inhibited
 Primary Isolation Media
• Brain heart infusion media
(BHI)
–Useful in the isolation of pathogenic
fungi from sterile specimens
–Can be supplemented with blood,
and antimicrobials
 Reproduction

• Asexual
– Yeast multiply by binary fission, budding,
pseudohyphae
– Molds reproduce asexually by fragmentation
of hyphae, or producing spores
– Spore - a reproductive particle, usually a single
cell, released by a fungus, alga, or plant that
may germinate into another fungi
Sexual
 HYPHAE

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 FUNGAL SPORES

11/26/2019 104
 Asexual Reproduction
Types of Spores
• Conidia
– Chlamydospore
– Blastospore/Blastoconidia
– Arthrospores
– Macro- and microconidia
– Phialides
– Sporangiospore
 Asexual Spores: Conidia
• Chlamydospore
– Thick-walled, resistant,
resting spores

– Produced by rounding up
and enlargement of hyphal
segments
– Candida albicans
 Asexual Spores: Conidia

• Blastoconidia
– Develop as daughter cell buds off from
parent cell and is pinched off
– Candida

• Pseudohyphae
– Candida
 Yeast
• Most reproduce by
budding (forming
blastoconidia); few by
binary fission
– Buds fail to detach
producing elongated
yeast cells
(pseudohyphae)
Pseudohyphae
 Asexual Spores: Conidia
• Arthroconidia
– Formed by
fragmentation of the
septate hyphae into
single rectangular or
barrel-shaped thick-
walled spores
– Coccidioides immitis
 Asexual Spores:
Macroconidia and Microconidia
• Macroconidia
– Large, multiseptate, club,
oval, spindle-shaped
– Cell wall is smooth or
echinulate
• Microcnidia
– Small, unicellular
– Round, elliptical. pyriform,
tear-shaped
 Asexual Spores:
Macroconidia and Microconidia
• Macroconidia
– Large, multiseptate, club,
oval, spindle-shaped
– Cell wall is smooth or
echinulate
• Microconidia
– Small, unicellular
– Round, elliptical. pyriform,
tear-shaped
 Macroconidia and Microconidia
Dermatophytes

• Microsporum
• Trichophyton
• Epidermophyton

– Dermatophytes are fungi that require keratin for

growth. These fungi can cause superficial
infections of the skin, hair, and nails.
 Dermatophytes
• The genus Trichophyton is capable of invading
the hair, skin and nails (All three so tri).

• The genus Epidermophyton involves the skin

and nails only (Not Hair).

• The genus Microsporum involves only the hair

and the skin (Not Nail).
• Microsporum
– Large
multicellular
spindle shaped
macroconidia;
microconidia
rarely produced
• Trichophyton
– Predominant forms
are microconidia;
macroconidia
uncommon
– Do not fluoresce on
Wood’s lamp
• Epidermophyton
– Club-shaped
Macroconidia only;
microconidia not
produced
 Primary Isolation Media
• Dermatophyte test medium
– Recovery of dermatophytes from hair, skin
and nails
– Useful as a screening medium
– Dermatophytes produce alkaline metabolites
which raise the pH and change the color of
medium into RED
 Asexual Reproduction
Types of Spores
• Conidia
– Chlamydospore
– Blastospore
– Arthrospores
– Macro- and microconidia
– Phialides
– Sporangiospore
 Asexual Spores:
Phialides (Secondary Branches)
• Aspergillus
• Penicillium
• Phialophora
• Annellids (Exophiala)

– A phialide/sterigma is an elongated and flask

shaped (where it gets its name) projection
rising from the vesicle in certain fungal groups
 Aspergillus

• Branching
septate hyphae
which bears
terminal
conidiophore

*
*
 Aspergillus
• Conidiophore
expands to large
inverted flask
shaped vesicle

*
 Aspergillus
• Vesicle is covered
with phialides on
upper half of the
vesicle

*
 Aspergillus
• Phialides contain
rough walled
conidia/conidiospor
es

*
*
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*
 Penicillium
• Hyphae are septated
with brush-like
conidiophore

• Conidiophores exhibit
branching to form
phialides/metulae

*
 Penicillium
• Brush-like
conidiophores
that give rise to
phialides from
which chain of
conidia arise
*
*
*
 Phialophora
• Flask-shaped
phialides with
cup-shaped
collarettes and
clusters of
conidia at the
end
 Annellids (Exophiala)

• Conidiophores are
long and cylindrical
(annellophores) with
tapered tip and
ringed by clusters of
conidia
 Asexual Reproduction
Types of Spores
• Conidia
– Chlamydospore
– Blastospore
– Arthrospores
– Macro- and microconidia
– Phialides
– Sporangiospore
 Asexual Spores:
Sporangiospores
• Asexual spores
contained in a sac-like
or sporangium
• Unique among fungi
with Nonseptated or
coenocytic hyphae
– Zygomycetes
– Rhizopus, Mucor
Absidia
 Mucor
• Nonseptated
hyphae with no
rhizoids
• The
sporangiophores
arise singly with
sac called
sporangium

*
*
 Mucor
• Sporangium
contains
sporangiospores

*
*
*
 Rhizopus
• Nonseptated hyphae
• With Rhizoids
• Sporangiophores
• Sporangium

*
*
 Asexual Reproduction
Types of Spores
• Conidia
– Chlamydospore
– Blastospore
– Arthrospores
– Macro- and microconidia
– Phialides
– Sporangiospore
 Sexual Reproduction
• Requires formation of specialized
structures so that fertilization or nuclear
fusion can occur

• Meiosis occurs with reduction division of 2

fertile cells, followed by merging of the
cells and nuclear fusion
 Sexual Reproduction
• A fungal sexual spore results from sexual
reproduction,
• 3 phases
– Meiosis – diploid nucleus become haploid
– Plasmogamy – fusion of cytoplasm resulting
to one organism with two nuclei
– Karyogamy – fusion of nuclei to form back
diploid state
• Plasmogamy
– A haploid nucleus of a donor cell (+) penetrates
the cytoplasm of a recipient cell (-)

• Karyogamy
– The (+) and (-) nuclei fuse to form a diploid
zygote nucleus

• Meiosis
– The diploid nucleus gives rise to haploid nuclei
(sexual spores)
 Types of Sexual Spores
• Ascospores
– Contained in a sac-like ascus
Note
 Types of Sexual Spores
• Zygospores
– Fusion of 2 identical cells arising from the
same hypha
 Zygomycota
• Aseptate hyphae
• Asexual spores: sporangiospores
• Sexual spores: zygospores
• Clinically important members:
– Rhizopus
– Mucor
– Absidia
 Types of Sexual Spores

• Oospores
– Fusion of cells from 2
separate non-identical
hyphae
 Types of Sexual Spores

• Basidiospores
– Contained in a club-
shaped basidium
• Teleomorphs
– The sexual stage of a fungus

• Anamorphs
– The asexual stage of a fungus
 Dimorphic Fungi
• Can be molds or yeasts

• Sporothrix schenckii
• Coccidioides immitis
• Histoplasma capsulatum
• Blastomyces dermatitidis
• Paracoccidioides braziliense