INTRODUCTION TO CLINICAL MYCOLOGY  Reproduce or multiply

o Asexually by budding or fission

GENERAL CHARACTERISTICS OF FUNGI
 A bud is sometimes called a blastospore or
 Eukaryotic; possess a true nucleus with a nuclear membrane blastoconidium (daughter cell)
and mitochondria  Budding is a mitotic process (asexual
process)
 Achlorophyllous
 It involves maturation of the bud to an
 Most of them are saprophytic
independent daughter cell
 Most are obligate aerobes that grow best at neutral pH
o Sexually by production of ascospores or
 Moisture is necessary for growth, but some forms survive in
basidiospores
dry conditions for extended lengths of time
 Steps in yeast budding:
 Thick cell walls composed of:
1. Lysis of the yeast cell wall (parent cell wall) so that a
o chitin, mannans, and sometimes cellulose
blastospore can form
o these properties allow the absorbing of specialized
2. Asthe blastospore enlarges, the nucleus of the
dyes used in identification
parent cell undergoes mitosis
 Reproduce sexually and/or asexually
3. One of the nucleus is passed into the blastospore
 More than 100,000 species of fungi are human pathogens
4. Septum forms and daughter cell breaks free through
 Exhibit great variation in size (from 3 microns to more or less 3 the process of fission or cytokinesis
feet), morphology, mode of reproduction, and methods of
dissemination
 Fungi are ubiquitous
 They are even found in sea water and in the arctic ice
 There is no escaping fungi  During fission, two cells of equal size are formed
 These cells continue to grow from the tips of the cell and
divide only after a medial fission is formed

Molds

 Filamentous fungi that are multicellular in structure

YEASTS VERSUS MOLDS  Macroscopically, most have a fluffy, fuzzy
or woolly or powdery appearance
Yeasts because of the formation of mycelia
 Single vegetative cells that  Mycelia (singular mycelium) are a mass of
typically form in culture media hyphae
as:  Hyphae are long, slender, branching
o smooth, moist cylindrical tubes and are considered the
o opaque or pasty basic structural units of molds
o creamy, bacterial-like  Aerial hyphae
colony without aerial o Extend above the
hyphae surface of the colony
 All yeasts are fungi and are responsible for
 Unicellular organism the fuzzy appearance
o Support the reproductive structures (fruiting bodies)
 Identification is based
that produce conidia
primarily on biochemical
testing  Vegetative hyphae
 Yeast produces catalase o Extend downward into the medium to absorb
nutrients
o Water exchanging portion
 Produce fluffy, wooly, or powdery colonies
on culture media

  Microscopic appearance often aids in the identification of  Sparsely septate hyphae
molds
o Different forms of hyphae
 Antler
 Racquet
 Rhizoids
 Pectinate
 Spiral

 Aseptate - absence of septa, has historically been used to

Different forms of hyphae describe the hyphae of Zygomycetes
 Microscopic examination of hyphae associated with
 Antler hyphae have swollen, branching tips Zygomycetes often reveals occasional septations; therefore,
that resemble moose antlers these hyphae are more correctly termed sparsely septate as
opposed to aseptate

Hyaline vs. Phaeoid Hyphae

 Racquet hyphae contain enlarged, club-shaped areas  Another characteristic useful in ID is pigmentation
 narrower on one end and wider-round on the  Hyaline (moniliaceous) hyphae
opposite end o non-pigmented or lightly pigmented

o darkly pigmented because of the presence of
 Pectinate hyphae are comb- melanin in the cell wall
like structures of hyphae  Depending on the amount of melanin present, the hyphae will
appear pale to dark brown or almost black

 Spiral hyphae are tightly

coiled hyphae

 Rhizoids are root-like DIMORPHISM AND POLYMORPHISM

structures Dimorphism
 Refers to the ability of some fungi to exist in two forms,
dependent on growth conditions
 Dimorphic fungi include a mold phase and a yeast or spherule
phase
YEASTS VERSUS MOLDS  Yeast or tissue state is seen in vivo or when the organism is
grown at 37oC
 Microscopic appearance often aids in the identification of  Mold phase is seen when the organism is grown at room
molds temperature (22oC to 25oC) in ambient air conditions
Septate vs. Sparsely Septate Hyphae  Examples of thermally dimorphic fungal species associated with
human disease:
 Frequently, when fungal hyphae are being described, they are o Blastomyces dermatitidis – blastomycosis
referred to as septate or sparsely septate o Coccidioides immitis – coccidiomycosis or
 Septate hyphae show frequently cross-walls occurring coccidioidomycosis
perpendicularly to the outer walls of the hyphae o Histoplasma capsulatum var. capsulatum –
 Sparsely septate hyphae have few cross-walls at irregular histoplasmosis
intervals o Paracoccidioides brasiliensis –
 Septate hyphae paracoccidioidomycosis
o Sporothrix schenckii – sporotrichosis (rose
gardener’s disease)
o Talaromyces marneffei – previously known as
Penicillium marneffei
Polymorphism Asexual Reproduction Through Asexual Spores

 Polymorphic fungi have both yeast and mold forms in the same  Spores are produced asexually from the mycelium
culture  Asexual reproduction give rise to an anamorph or imperfect
 This characteristic occurs despite growth conditions and is best fungi or fungi imperfecti
observed in Exophiala spp., in which the yeast phase is typically  Asexual reproduction involves only mitosis with nuclear and
observed initially, followed by the mold phase as the colony cytoplasmic division
ages o carried out by specialized fruiting structures known as
conidiogenous cells (cells that produce conidia)
REPRODUCTION o Conidia contains all the genetic material necessary
Spores to create a new fungal colony
 Formation of conidia or conidiospores is at the tips or sides of
Functions primarily for reproduction hyphae following mitosis
 Conidia(sing. conidium)
 Functionally similar to the seeds of higher plants
o Asexual spores
 Formed from aerial mycelium in different ways depending on produced either
the species singly or multiply
 The manner in which spores are produced is important in the in long chains or
identification of fungi clusters by
 There are two types of spores: specialized
o Sexual spores (sexual reproduction) vegetative
o Asexual spores (asexual reproduction) hyphae known as
conidiophores
Fungi produce spores also for survival
o Conidiophores
 Resistant to adverse conditions and germinate when conditions may branched into secondary segments, known as
for growth are favorable phialides, which then produce the conidia.
 Thallospores
Sexual Reproduction Through Sexual Spores o Derived from cells of thallus or body of fungus
 Fungi that exhibit a sexual reproduction give rise to a telemorph Some spores develop directly from vegetative hyphae
or perfect fungi
 This requires the joining of two compatible nuclei, so that  Arthroconidia (Arthrospores)
fertilization or nuclear fission can occur followed by meiosis o Formed directly from hyphae by fragmentation at
points of septation
Types of specialized spores o When mature they appear as jointed, rectangular or
barrel-shaped, thick-walled cells
 Ascospore
o May occur naturally by the action of wind, soil
o Spore contained in a sac-like structure called ascus
movement, or insects
 Basidiospore
o Adjacent – example is Geotrichum – arthroconidia are
o Spore formed at the end
referred to as oidia due to their flat ends
of club-shaped structures
o Alternate – example is Coccidioides immitis
called basidium

 Blastoconidia (Blastospores)
 Zygospores o Developed as the daughter cell buds off the mother
o Formed by conjugation between two morphologically cell and is pinched off from portions of the mother
identical cells arising from the same hypha cell
 Oospores
o Formed by
heterogeneous
fertilization or
fusion between
to separate,
non-identical
hyphae
 o May elongate to form pseudohyphae which can align FUNGAL CELL
from in an end to end manner, resembling true
hyphae  It is very similar in many ways to the cells found in higher forms
o Pseudohyphae are constricted at the septa and of life
branches develop at the septation  Septate fungi are only septate from a morphological point of
o True hyphae are not constricted at the septa view
 Chlamydoconidia (Chlamydospores)  Physiologically, they are coenocytic because the septa have
o Formed by enlargement of a hyphal cell in pores in them large enough to allow nuclei to pass through
which there is a concentration of protoplasm  One outstanding characteristic is that fungal cells have very
and nutrient material thick cell walls
o Arrangement of chlamydospores:  Fungal cell wall generally contains 5-10% protein
 Terminal – formed at the hyphal tip  In the cell walls of yeasts, the major components are
 Sessile – hyphal sides carbohydrate polymers, such as glucans and mannans, which
 Intercalary – within hyphal strands usually constitute 50-60% of the cell wall
 All typical fungi generally has a very high content of chitin in
the cell wall
 Chitin, made of N-acetylglucosamine, is one of the toughest
 Conidia
biologically produced materials
o Some conidiophores terminate in swollen
vesicles from which short phialides form and in
turn produce radiating chains
o This is a characteristic of Aspergillus TAXONOMY
o Some conidiophores form a branching structure
CLASSIFICATION OF FUNGI
called penicillus, in which each branch
terminates in secondary branches (metulae)  There are over 100,000 named fungal species and an
and phialides, from which chains of conidia estimated 1 to 10 million undiscovered species
are borne  Most etiologic agents of clinical infections are found in
o This type of sporulation is the characteristic four groups of fungi:
of Penicillium o Phylum Glomeromycota
o Some fungi may produce conidia of two sizes: o Phylum Ascomycota
 Microconidia – small, unicellular, o Phylum Basidiomycota
round, elliptical or pyriform (pear- o Phylum Deuteromycota (Fungi Imperfecti)
shaped)
 Macroconidia – large, usually PHYLUM GLOMEROMYCOTA
multiseptate, and club- or spindle-
shaped  The traditional Zygomycota have undergone taxonomic
changes
 The most clinically significant species belong to the phylum
Glomeromycota in the order Mucorales
 Mucorales include the genera Lichtheimia (formerly Absidia),
Mucor, Rhizomucor, and Rhizopus
 Members of the order are rapidly growing organisms normally
found in soil, and are often opportunistic pathogens in
immunocompromised host
 Mucorales generally produce profuse, gray to white, aerial
mycelium characterized by the presence of hyaline, sparsely
septate hyphae
 Sporangiospores  Asexual reproduction is characterized by the presence of
o Some spores are borne in a sac-like structure called a sporangiophores and sporangiospores
sporangium  Sexual reproduction, resulting in the production of
o The sporangium is borne on a tip of a supporting zygospores, occurs in some, but these structures are not
structure called the sporangiophore or aseptate routinely seen in clinical laboratories
hypha
o The spores contained are called sporangiospores PHYLUM ASCOMYCOTA
o Typically seen among Zygomycetes
 Fungi associated with the class Ascomycetes are characterized
by the production of sexual spores called ascospores
 They are usually identified on the basis of characteristic
asexual structures
  Examples include Microsporum spp., Trichophyton spp., and  Specimen placed directly on a microscopic
Pseudallescheria boydii slide
 One to two drops of 10-20% KOH is added
 KOH clears away much of the tissue debris
but does not break down fungal structures
 Add coverslip, press it down, and observe
specimen microscopically
 Appearance of the organism in tissue, which is considered an
important tool in diagnosis of a fungal disease
o Stained Histology Slide
 Most fungi are larger than 3 microns
PHYLUM BASIDIOMYCOTA  This makes fungi much easier to see than
bacteria in stained slides
 Sexual function results in the formation of a club-shaped organ  Most commonly used stains for observing
called basidium, on the surface of which are borne the four fungi in tissue are periodic acid-Schiff (PAS)
meiotic products, basidiospores and Gomori’s methenamine silver (GMS)
 Asexual spores (conidia) are borne externally at the tips of stain
hyphae
 Clinically significant WAYS FUNGI MAY BE OBSERVED IN TISSUE
basidiomycetes are few
Yeast Cells
and the only known
pathogen is Filobasidiella  One of the most common forms in which one sees fungi in
neoformans, the perfect tissue
form (teleomorph) of  Gomori methenamine silver (GMS) stain is particularly
Cryptococcus neoformans valuable for visualizing yeast and other forms of fungi in tissue
var. neoformans  Yeast cells may be intracellular, such as Histoplasma
DEUTEROMYCOTA (FUNGI capsulatum, which is one of the few fungi that can be
IMPERFECTI) classified as intracellular parasite
 In cryptococcosis, most yeast cells have a large distinguishing
 This division contains the largest number of organisms that are capsule; Cryptococcus neoformans apparently is the only
causative agents of mycoses, including cutaneous, pathogenic encapsulated yeast
subcutaneous, and systemic disease  Many yeast cells in tissue exhibit buds
 Fungi which lack  A large neck (broad base) between the mother cell and the
sexual reproduction bud is a diagnostic feature in cases of blastomycosis
 Asexual reproduction  Some have multiple buds, e.g. yeast cells of
is primarily through
conidia
 Organisms in this
group are identified
on the basis of
characteristic asexual reproductive structures
 Examples include Epidermophyton spp., Sporothrix spp., and
Candida spp.

CLINICAL MYCOLOGY

FACTORS IN IDENTIFYING MYCOTIC DISEASES

 In almost all cases, fungal diseases are chronic

o Take months or years to develop
o Major exceptions are primary pulmonary form of
histoplasmosis and coccidiomycosis
 History taken from the patient and clinical picture
 Factors which predispose humans to fungal diseases
o Diabetes mellitus, cancer, alcoholism, AIDS
o Recent surgery
o Administration of immunosuppressive agents and/or
antibiotics
 Appearance of the organism in tissue, which is considered an
important tool in diagnosis of a fungal disease
o Direct examination
 Granules

 Tightly packed masses of hyphae or filaments which are

surrounded by a tough outer rind
 May be grossly visible, 0.5 to 10 mm in diameter
 Microscopically, they contain either fine, delicate filaments or
thick, septate hyphae

Fission Bodies

 Spherical, dematiaceous structures

Sporangia  Neither bud nor produce hyphae
 Division is by splitting down the middle
 Large, sac-like structures
 Some medical mycologists call these structures, sclerotic
which fill with spores as they mature
bodies or Medlar bodies
 Spores contained in these sacs are called sporangiospores or
 Characteristics of the disease known as
endospores
 Some books refer to them as spherules

Hyphae Combination of Yeast Cells and Hyphae

 Usually 5 to 10 microns in diameter  Seen in candidiasis only
 Sparsely septate hyphae (coenocytic or aseptate) versus
septate
 Hyaline versus dematiaceous