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
