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BASIC MYCOLOGY

MICROBIOLOGY LECTURE SERIES

LUZ GREGORIA VELASCO. MD

FUNGI
Eukaryotic; most are nonmotile and possess a rigid wall
Nonphotosynthetic
<400 species are medically important

<50 species cause >90% of the fungal infections of humans and other animals
Reside in nature and essential in breaking down and recycling organic matter
Contribute to the production of food and spirits (cheese, bread, beer)

Provide useful bioactive secondary metabolites (antibiotics- penicillin;

immunosuppressive drugs cyclosporine)


Phytopathogens

FUNGI

Each fungal cell has at least one nucleus with a nuclear membrane, ER,

mitochondria and secretory apparatus


Most are obligate or facultative aerobes
Chemotrophic

MYCOSES

Fungal infections
Most pathogenic fungi are exogenous

Highest incidence: candidiasis, dermatophytosis

MAJOR MYCOSES
CATEGORY
Superficial

Cutaneous

MYCOSIS

CAUSATIVE FUNGAL
AGENTS

Pityriasis versicolor
Tinia nigra
White piedra
Black piedra
Dermatophytosis

Malassezia species
Hortaea werneckii
Trichosporon sp.
Piedraia hortae
Microsporum species,
Trichophyton species,
Epidermophyton floccosum
Candida albicans and other
Candida species

Candidiasis of skin,
mucosa or nails

MAJOR MYCOSES
CATEGORY
Endemic
(primary,
systemic)

MYCOSIS
Sporotrichosis
Chromoblastomycosis
Mycetoma

Phaeohyphomycosis

CAUSATIVE FUNGAL
AGENTS

Sporothrix schenckii
Phialophora verrucosa
Fonsecaea pedrosoi and others
Pseudallescheria boydii,
Modurella mycetomatis, and
others
Exophiala, Bipolaris, Exserohilum,
and other dematiaceous
molds

MAJOR MYCOSES
CATEGORY
Opportunistic

MYCOSIS
Systemic candidiasis
Cryptococcosis
Aspergillosis
Hyalohyphomycosis

CAUSATIVE FUNGAL
AGENTS

Candida albicans and other


Candida species
Cryptococcus neoformans and
Cryptococcus gattil
Aspergillus fumigatus and other
Aspergillus species
Species of Fusarium,
Paecilomyces,Trichosporon, and
other hyaline molds

MAJOR MYCOSES
CATEGORY
Opportunistic

MYCOSIS
Phaeohyphomycosis

Mucormycosis
(zygomycosis)

CAUSATIVE FUNGAL
AGENTS

Chladophialophora bantiana,
species of Alternaria,
Cladosporium, Bipolaris,
Exserohilum and other
dematiaceous molds
Species of Rhizopus,
Lichtheimia, Cunninghamella, and
other zygomycetes

MAJOR MYCOSES
CATEGORY
Opportunistic

MYCOSIS
Pneumocystis pneumonia
Penicillosis

CAUSATIVE FUNGAL
AGENTS

Pneumocystis jiroveci
Penicillum marneffei

GENERAL PROPERTIES AND CLASSIFICATION OF


FUNGI
Fungi grow in two basic forms:

1. YEASTS- grow as single cells that reproduce by asexual budding


2. MOLDS- production of multicellular filamentous colonies
branching cylindric tubules (hyphae) 2-10 mm
mycelium- mass of intertwined hyphae that accumulates during
active growth

some hyphae are divided into cells by cross-walls (septa),


typically form at regular intervals during hyphal growth

GENERAL PROPERTIES AND CLASSIFICATION OF


FUNGI

A - coenocytic hypha
B - septate hypha
C - septum

GENERAL PROPERTIES AND CLASSIFICATION OF


FUNGI
Vegetative or substrate hyphae penetrate the surrounding

medium, anchor the colony and absorb the nutrients


Aerial hyphae project above the surface of the mycelium; usually

bear the reproductive structures of the mold


When a mold is isolated from a clinical specimen, its growth rate,

macroscopic appearance and microscopic morphology are usually


sufficient to determine its genus and species
Most helpful phenotypic features: ontogeny and morphology of the

asexual reproductive spores or conidia

HYPHAE
Coenocytic /aseptate vs.
Septate
Hyaline vs.
dematiaceous
Fine vs. broad

dichotomously
branching
septate hyphae

nonseptate hyphae

GENERAL PROPERTIES AND CLASSIFICATION OF


FUNGI

CONIDIA asexual reproductive spores (MITOSPORES) produced either

from the transformation of a vegetative yeast or hyphal cell or from a


specialized conidiogenous cell, which may be simple or complex and elaborate
May be formed on specialized hyphae (conidiophores)
MICROCONIDIA small
MACROCONIDIA large or multicellular

Microconidia
Singly or in clusters

Macroconidia

GENERAL PROPERTIES AND CLASSIFICATION OF


FUNGI
YEASTS single cells, usually spherical to ellipsoid in shape, 3-15 mm
Most reproduce by budding
Some produce buds that characteristically fail to detach and become elongated

(pseudohyphae chain of elongated yeast cells)


Colonies usually soft, opaque, 1-3 mm in size, cream-colored
Yeast species are identified on the basis of physiologic tests, and a few key

morphologic differences

YEAST CELLS

Budding yeast
cells surrounded
by clear capsules

encapsulated spherical yeast cells

cigar to oval shaped yeast cells

broad based bud

GENERAL PROPERTIES AND CLASSIFICATION OF


FUNGI
Thermally dimorphic (form different structures at different temperatures)
Essential rigid cell wall (determines shape and protects from osmotic

and environmental stress)


Composed largely of carbohydrate layers (long chains of polysaccharides),

glycoproteins and lipids


Sugar polymers: Chitin (b-1,4-linked N- acetylglucosamine), glucans (b-1,3-glucan, b1,6-glucan, mannans (polymers of mannose- a-1,6-mannose)

GENERAL PROPERTIES AND CLASSIFICATION OF


FUNGI
Essential rigid cell wall
During infection, fungal cell walls exert important pathobiologic properties-

surface component mediate attachment to host cells


bind to pattern recognition receptors on host cell membranes
cell wall glucans and other polysaccharides activate the complement cascade and
provoke an inflammatory reaction
release immunodominant antigens that may elicit cellular immune responses and
diagnostic antibodies
dematiaceous brown or black pigment to the fungal colony (melanized cell
walls)

GENERAL PROPERTIES AND CLASSIFICATION OF


FUNGI

Most fungi are obligate aerobes, some are facultative anaerobes, but

none are obligate aerobes


All fungi require a preformed organic source of carbon

Natural habitat of most fungi: environment, except Candida albicans,

which is part of the normal human flora

GENERAL PROPERTIES AND CLASSIFICATION OF


FUNGI
FEATURE

FUNGI

BACTERIA

Diameter

Approximately 4mm (Candida) Approximately 1mm


(Staphylococcus)

Nucleus

Eukaryotic

Prokaryotic

Cytoplasm
Cell Membrane

Mitochondria & ER present


Sterols present

Cell Wall Content

Chitin

Mitochondria & ER absent


Sterols absent (except
Mycoplasma)
Peptidoglycan

GENERAL PROPERTIES AND CLASSIFICATION OF


FUNGI
FEATURE

FUNGI

BACTERIA

Spores

Sexual and asexual spores for Endospores for survival, not for
reproduction
reproduction

Thermal Dimorphism Yes (some)

No

Metabolism

Many do not require organic


carbon, many obligate anaerobes

Require organis carbon, no


obligates anaerobes

CLASSIFICATION ACCORDING TO COLONY TYPE AT


DIFFERENT TEMPERATURES

MONOMORPHIC OR MONOPHASIC produce one type of

colony at both room and incubation temperature


Ex.

Cryptococcus neoformans yeast only


Aspergillus species filamentous only

Candida albicans yeastlike colony

COLONIES PRODUCED BY FUNGI


YEAST COLONIES
soft, pasty type of colony like
yeast produced by fungi which
remains at yeast form at both
4o C and 37oC

COLONIES PRODUCED BY FUNGI


YEASTLIKE COLONIES
soft, pasty type of colony
produced by fungi which have the
ability to form pseudomycelia
(false hypha)
- Pseudohypha consist of
elongated buds remaining attached
to the parent cell despite repeated
division.

COLONIES PRODUCED BY FUNGI


FILAMENTOUS OR MOLD
COLONY
a mold type of colony with an
aerial mycelium that appears
cottony, wooly, powdery or
granular.
- true mycelia are produced by
spores which germinate to form
branching hyphae

CRYPTOCOCCUS NEOFORMANS
ROOM TEMPERATURE
CULTURE

37oC

Off white, mucous colonies on Sabourauds medium after 5 to 10 days

ASPERGILLUS SPP.
ROOM TEMPERATURE
CULTURE

37oC

Green colony after 5 to 10 days on Sabouraud medium. Colorless underside.

CANDIDA ALBICANS
ROOM TEMPERATURE
CULTURE

37oC

White colonies incubated on Sabouraud medium for 5 to 7 days.

CLASSIFICATION ACCORDING TO COLONY TYPE AT


DIFFERENT TEMPERATURES
DIMORPHIC OR DIPHASIC produce a yeast-like colony at 37 C

or incubation temperature (YEAST PHASE) and a filamentous type of


colony at room temperature (MYCELIAL PHASE)

Ex.

Sporothrix schenckii
Histoplasma capsulatum
Blastomyces dermatitidis

SPOROTHRIX SCHENCKII
ROOM TEMP
CULTURE

Dark, greasy-looking culture after 1 to 2


weeks incubation on Sabouraud medium

37oC

White to tan yeast colonies after 1 1 to 3 weeks


incubation on brain heart infusion agar.

HISTOPLASMA CAPSULATUM
ROOM TEMP
CULTURE

White, innocent looking(but dangerous)


colony after 1 to 3 weeks incubation on
Saboraud medium.

37oC

White to tan yeast colony after 1 to 3 weeks


incubation on brain heart infusion agar

BLASTOMYCES DERMATITIDIS
ROOM TEMP
CULTURE

Colony after 1 to 3 weeks incubation on


Sabouraud medium. white colony, brown to
tan underside

37oC

Rough, dry, heaped-up yeast colony after 1 to


3 weeks incubation on brain heart infusion
agar

TAXONOMY

PHYLUM GLOMERULOMYCOTA, ORDER MUCORALES

PHYLUM ASCOMYCOTA (Ascomycetes)


65% of the known fungi, 85% of the human pathogens

PHYLUM BASIDIOMYCOTA (Basidiomycetes)

TAXONOMY
PHYLUM GLOMERULOMYCOTA, ORDER MUCORALES

Sexual reproduction results in a ZYGOSPORE; asexual reproduction occurs via

SPORANGIA
Vegetative hyphae sparsely septate

Rhizopus, Lichtheimia, Mucor, Cunninghamella

TAXONOMY
PHYLUM ASCOMYCOTA (Ascomycetes)
Sexual reproduction involves a sac (ascus) in which karyogamy and meiosis

occurs ASCOSPORES; asexual reproduction occurs via CONIDIA


(+) septate hyphae
Yeasts (Saccharomyces, Candida); molds (Coccidioides, Blastomyces, Trichophyton)

PHYLUM BASIDIOMYCOTA (Basidiomycetes)


Sexual reproduction results in dikaryotic hyphae and 4 progeny

BASIDIOSPORES supported by a club-shaped basidium; no asexual spores


Hyphae with complex septa; Mushrooms, Cryptococcus

DIAGNOSIS

Species of a clinical isolate can be identified by molecular or phenotypic

methods (signature DNA sequences, morphology of reproductive


structures, physiologic properties)

SEXUAL SPORES:
1. zygospores single large spores with thick walls
2. ascospores formed in a sac (ascus)

3. basidiospores formed externally on a tip of a pedestal (basidium)


FUNGI IMPERFECTI - Fungi that do not form sexual spores are

imperfect

ASCOSPORES

produced within an
enlarged cell by
nuclear fusion; enclosed
in a sac called an
ASCUS or
ASCOCARP.
Typically, there may be

4-8 spores within an


ascus.

ASCOSPORES

ASCOCARP

ASEXUAL SPORES
arise by the differentiation of spore bearing hyphae without nuclear fission
most fungi of medical interest propagate asexually by forming asexual

spores (conidia) from the sides or ends of specialized structures

1. arthrospores
2. chlamydospores
3. blastospores
4. sporangiospores

ARTHROSPORES fragmentation or segmentation of preformed


mycelium results in the production of rectangular, thick-walled
spores which are uniformly sized
Ex. Geotrichum
Coccidioides immitis

Arthroconidia
formation

(A) produced by the


breaking down of a hyphal
strand(B) into individual
rectangular units.

CHLAMYDOSPORES large, round, thick-walled resting cells in


a septate hypha or at the terminal ends; formed when the
cytoplasm of these hypha become concentrated and enlarge in a
diameter greater than that of the rest of the hypha; may be
intercalary, terminal or lateral.
Ex. Candida albicans

CHLAMYDOSPORES

BLASTOSPORES
formed by a simple
budding process from
cells of the mycelium.
Budding may be in:
1)one cell to one spore
basis
Blastomyces dermatitidis
2) multispore basis
Paracoccidioides braziliensis

SPORANGIOSPORES

produced on a specialized
hypha, inside a round
container known as
SPORANGIUM.
When the sporangium ruptures, the sporangiospores scatter and leave the
thin walled sporangium in place or it may dissolve

GROWTH AND ISOLATION OF FUNGI


Saborauds agar traditional mycological medium
Contains glucose and modified peptone (pH 7.0)
Does not readily support the growth of bacteria

Inhibitory mold agar

MYCOTOXINS
Poisonous substances produced by fungi
Cause acute or chronic intoxication and damage
Produced by mushrooms (Amanita sp.) mycetismus

Aflatoxin Aspergillus flavus

ANTIFUNGAL THERAPY
POLYENES Amphotericin B, Nystatin
Bind to ergosterol in the cell membrane

FLUCYTOSINE pyrimidine analog


AZOLES interfere with the synthesis of ergosterol
ECHINOCANDINS inhibit the synthesis of cell wall b-glucan

GRISEOFULVIN interferes with microtubule assembly


TERBINAFINE allylamine drug; blocks ergosterol synthesis by

inhibiting squalene epoxidase

CONIDIA ACCORDING TO SHAPE

FUSIFORM spindle-shaped
CLAVATE club-shaped

MURIFORM multiseptate, both transverse and longitudinally

CONIDIA ACCORDING TO ARRANGEMENT


SESSILE AND LATERAL - develops directly on the side of

the hypha with no conidiophore or stem


EN GRAPPE - clustered
PEDUNCULATE - develop from the end of a short

conidiophore