MYCOLOGY AND VIROLOGY Floccose
MODULE 1 UNIT 1:
BASIC CONCEPTS IN MYCOLOGY
DEFINITION OF TERMS
Anamorph The asexual form of a fungus
Annellide A specialized conidiogenous
cell from which a succession
of spores is produced and
which has a column of apical
scars at its tip
Assimilation Ability of a fungus to use a
specific carbon or nitrogen
source for growth.
Aseptate Without cross-walls or septa.
Conidiogenesis The process of conidium
formation. Two basic
methods, thallic [in which an
existing hyphal cell is
converted into one or more
conidia] and blastic [in which
conidia are produced as a
result of some form of
budding process]
Conidium An asexual (mitosis only)
propagule that forms on the
side or the end of the hypha
or conidiophore. It may
consist of one or more cells. It
is always borne externally, ie.,
not enclosed in a saclike
structure such as sporangium.
If a fungus produces two types
of conidia, those that are
small and usually singled cell
are referred to as
microconidia, whereas the
larger macroconidia are
usually segmented into two or
more cells
Dematiaceous Darkly pigmented.
Fermentation Ability of a fungus to utilize a
specific carbohydrate in the
presence of other organic
compounds, resulting in the
production of gas. Acid
production (color change) may
simply indicate that the
carbohydrate has been
assimilated. All carbohydrates
fermented by a fungus are
also assimilated, but many
compounds that are
assimilated are not necessarily
fermented
Colonies with a cotton-like
texture.
Fusiform Term used to describe spores
with a spindle-like shape
Glabrous Colonies with wax-like texture
Heterothallic A self-sterile fungus; sexual
reproduction cannot take
place unless two compatible
mating strains are present
Homothallic A self-compatible fungus;
sexual reproduction can take
place within an individual
strain
Hyaline Colorless, transparent,
transluscent
Hypha (pl., hyphae) One of the individual filaments
that make up the mycelium of
a fungus.
Mold A filamentous fungus
Moniliaceous Hyaline or lightly colored
Mycelium A mass of branching filaments
which make up the vegetative
growth of a fungus
Oligokaryotic A cell with several nuclei
Pauciseptate Having few septa
Pseudohypha (pl., A chain of yeast cells which
pseudohyphae): have arisen as a result of
budding and have elongated
without becoming detached
from each other, forming a
hypha-like filament. Unlike a
true hypha, the connection
between adjacent
pseudohyphal cells shows a
marked constriction.
Rhizoid A short branching hypha that
resembles a root
Septate having cross-walls or septa
Species complex A monophyletic clade of
species with equivalent clinical
relevance
Sporangiophore A specialized hypha upon
which a sporangium develops
Sympodial Term used to describe the
development of a single
conidium at successive sites
along a lengthening
conidiogenous cell.
Teleomorph The sexual form of a fungus
Thallic One of the two basic forms of
conidiogenesis
Thallus The vegetative growth of a
fungus
Yeast A unicellular, budding fungus
 INTRODUCTION CLASSIFICATION OF THE FUNGI
 There are at least 135,000 named species of fungi with  Eukaryotes are divided into five monophyletic lineages or
an estimation of undescribed species ranges from 1 supergroups as proposed by Adl et.al.
million to more than 10 million, and calculated that 1. SAR (one clade consists of three groups viz.,
about 1,000 to 1,500 new species are described each Stramenopiles, Alveolata, and Rhizaria)
year. 2. Archaeplastida
 From the named species, fewer than 500 have commonly 3. Excavata
been associated with human and zoonotic diseases, and 4. Amoebozoa
no more than 50 causes opportunistic infections. 5. Opisthokonta — True fungi
 Importance of  Parafungi or pseudofungi— organisms that are not fungi
1. Increasing number of ubiquitous environmental sensu strictu (eg., Rhinosporidium seeberi) that share
molds implicated as opportunistic pathogens capable fungal-like morphological features with the true fungi.
of producing serious diseases among debilitated or  Fungi Imperfecti also termed “form-division
immunocompromised hosts. Deuteromycota” — fungi that reproduced asexual type
2. One of the leading causes of nosocomial infections. only (sporogenesis), while perfect fungi are capable of
3. Often mistaken for as bacterial infection that both sexual (teleomorph) and asexual (anamorph) type
produces fatal consequences. of reproduction.
4. Increase morbidity (travel to an endemic area or a
fungus that exists as part of indigenous flora of the BASIC SCHEME OF HIERARCHICAL ORGANIZATION OF THE
local population) KINGDOM FUNGI
5. Aging population

BRIEF HISTORICAL BACKGROUND

 35,000 BC: Fungi were used as an antiseptic and
anesthesia due to the “magical & spiritual” properties.
 Middle Ages: People were convinced that association
with fungi will entail the formation of diseases. People
believed that fungi were “the work of the devil”.
 Renaissance period: Fungi were plants with no fruit nor
seed.
 18th Century: Birth of the 1st mycological studies. KINGDOM FUNGI
Pier Anton Micheli— founder of modern mycological Seven phyla that constitute the true fungi
studies. Mycology was separated from botany. 1. Ascomycota Classified as the
 Mid-20th Century: Fungi were recognized as potential 2. Basidiomycota subkingdom Dikarya

causative agents of diseases that are usually fatal in 3. Blastocladiomycota

nature. 4. Chytridiomycota
5. Glomeromycota (formerly Zygomycota)
TAXONOMY AND NOMENCLATURE OF FUNGI  Consists of four subphyla incertae sedis.
 Taxonomy (Gk.: taxis = arrangement; anomia = method):  Subphylum Mucormycotina accommodates
branch of science that deals with systemic biological Mucorales that includes the genera Lichtheimia
classification of all living organisms and divided into three (formerly Absidia), Mucor, Rhizomucor, and
disciplines viz., classification, nomenclature, and Rhizopus.
identification.  Subphylum Entomophthoromycotina has been
 The scientific names of fungi are subject to the created for the Entomophthorales that includes the
International Code of Nomenclature (ICN) that was genera Basidiobolus and Conidiobolus — agents of
adopted in 2011. subcutaneous infections.
 In 2011, a breakthrough in the development of “One 6. Microsporidia
Fungus, One Name” concept recommended the 7. Neocallimastigomycota
discontinuation of the dual nomenclature system for
fungi with anamorphic and teleomorphic forms. Effective Phylum Basidiomycota
January 1, 2013 the system of permitting separate  Most members are septate, filamentous thallus, but
anamorph and teleomorph names ended; and assigned some are atypical yeasts.
priority to the oldest genus (species) name  Basidiospores (haploid) — produced via sexual
independently of whether the organism was originally reproduction of a generative cell (basidium).
described as the anamorph or the teleomorph.
 The most prominent are the basidiomycetous yeast with 3. Hypocreales: contains the teleomorphs of the
anamorphic stages belonging to the genera: anamorphic genus Fusarium.
1. Cryptococcus 4. Capnodiales
 Polyphyletic, belong to subphylum 5. Chaetothyriales Belonging to class
Eurotiomycetes or
Agaricomycotina, class Tremellomycetes, and 6. Microascales Sordariomycetes
order Tremellales and(or) Filobasidiales 7. Pleosporales Teleomorphs of melanized
fungi
2. Malassezia 8. Ophiostomatales
3. Trichosporon
 Asexual reproduction is variable, some produce MORPHOLOGICAL CHARACTERISTICS OF THE FUNGI
spores like those of the Ascomytoca. Multicellular fungi
 Most filamentous basidiomycetes are wood-  Hypha: basic structural unit consists of a chain of
rotting fungi or obligate plant pathogens. multinucleate, tubular, filament-like cells.
 Most multicellular fungi in their vegetative state, consists
Phylum Ascomycota of a mass of branching hyphae called mycelium or
 Contains ~50% of all named fungal species and accounts thallus.
for ~80% of fungi of medical importance.  Each hypha has a rigid cell wall and increases in length as
 Sexual reproduction leads to the development of haploid a result of apical growth with mitotic cell division.
spores, termed ascospores which are produced in a sac  In a more primitive fungus, the hyphae remain aseptate
like structure called ascus. Ascocarps or ascomata (coenocytic).
contains numerous asci.  In a more advanced groups, the hyphae are divided into
 Asexual reproduction leads to conidiation from a compartments or cells by the development of more or
generative conidiogenous cell. less frequent cross-walls, termed septa.— septate
 Three medically important subphylum: hyphae.
1. Taphrinomycotina contains the genus  Fungi that exist in the form of microscopic multicellular
Pneumocystis formerly classified under kingdom mycelium are commonly referred to as molds.
Protozoa.
2. Saccharomycotina contains the class Unicellular fungi
Saccharomycetes belongs to order  Exists in the form of independent single cells propagate
Saccharomycetales by budding out similar cells from their surface.
3. Pezizomycotina contains two classes:  The bud may become detached from the parent cell or it
Eurotiomycetes and the Sordariomycetes may remain attached and itself produce another bud,
leading to the production of a chain of cells or described
Phylum Ascomycota— Class Saccharomycetes as loosely arrangement of budding cells — yeasts.
 Characterized by vegetative yeast cells which proliferate  Under certain conditions, continued elongation of the
by budding or fission. parent cell before it buds out results in a chain of
 Do NOT produce ascomata, the ascus being formed by elongated cells, termed pseudohypha.
direct transformation of a budding vegetative cell, by  Compare to true hypha, the connection between
“mother-bud” conjugation, or by conjugation between adjacent pseudohyphal cells shows a marked
two independent single cells. constriction.
 Members include the anamorphic stage belonging to
genus Candida containing ~200 anamorphic species and Dimorphic fungi
has teleomorphs in more than 10 genera.  Medically important fungi that change their growth form
1. Clavispora as a part of the process of tissue invasion.
2. Debaryomyces  Dimorphic pathogens change from a multicellular mold
3. Issatchenkia form (25-30°C) in the natural environment to a budding,
4. Kluyveromyces single-celled yeasts form (35-37°C) in tissue under the
5. Pichia influence of temperature— thermal dimorphism.
1. Histoplasma capsulatum
Phylum Ascomycota — Class Eurotiomycetes 2. Blastomyces dermatitidis
 Sexual reproduction leads to the formation of ascomata 3. Coccidioides immitis
containing asci with ascospores. 4. Paracoccidioides brasiliensis
 Contains seven orders that are pathogenic to humans 5. Sporothrix schenckii
viz.:
1. Onygenales: contains teleomorph of the
dermatophytes and numbers of dimorphic system
pathogens ie., H. capsulatum and B. dermatitidis.
2. Eurotiales: includes teleomorphs of anamorphic
genera ie., Aspergillus and Penicillium.
 FUNGAL CYTOLOGY FUNGAL PHYSIOLOGY
Nutritional Type is Chemoheterotrophic (Lacks Chlorophyll)
 Require preformed organic carbon compounds for
nutrition.
 Obtain their nourishment by secreting enzymes into the
external substrate and by absorbing the released
nutrients through their cell wall.
 Capable of growing on dead (saprophytic) and living
organic matter.

Cell Wall Motility and Atmospheric Requirements

 Rigid, mostly composed of 90% polysaccharides eg.,  Usually nonmotile except for the phylum
chitin, glucan, chitosan, galactosan, and mannan; 10% Chytridiomycota and species of Rhizophidium.
proteins and glycoproteins in various combinations.  Mostly aerobic, some are facultative anaerobic.
 Chitin — the major carbohydrate consist of repeating
monomers of N-acetyl-glucosamine (NAG) which Lysine Synthesis
provides shape and protection from osmotic lysis  Utilize α-aminodiphate pathway.
(unaffected by some antibiotics).  Precursor for penicillin and lysine, also for the
 Functions: development of antifungal drugs.
1. Barrier between fungal cell and its external
environment. Reproduced by Means of Microscopic Propagules either
2. Binding site for some enzymes Spores or Conidia.
3. Possesses antigenic properties which allow  Conidium (pl. conidia) describes the propagules that
interaction with other organisms result from an asexual process (mitosis only) and
generally short-lived propagules.
Cell Membrane  Most fungi are capable of sexual reproduction (involving
 Composed of structurally arranged phospholipids in two- meiosis, preceded by fusion of the nuclei of two cells).
layered configuration scattered randomly.  Some species are homothallic (self-fertile) and able to
 Ergosterol — major component that regulates solute form sexual structures between different cells within an
intake and secretion (transport system) through selective individual thallus.
permeability and serves as the target of antifungal drugs  Most are heterothallic that do not form their sexual
like Nystatin. structures unless two capable isolates come into contact
 Facilitates biosynthesis of cell wall and capsular material. — once two compatible nuclei have fused, meiosis can
 Protects the cytoplasm. occur leading to the production of sexual Spores.
 In some cases, spores are produced in millions in
Capsule macroscopic “fruiting bodies” such as mushrooms.
 External coating located outside or covering the cell wall
and found only in certain fungi. FUNGI VS. BACTERIA
 Composed of amorphous polysaccharide. Characteristics Fungi Bacteria
 May influence growth of fungus by preventing Polysaccharides (ie.,
dissociation of buds and dispersion of yeast. Chitin, Glucan, Polysaccharide
 Virulence factor Cell wall
Mannan, Galactosan, Peptidoglycan
Chitosan
Cytoplasm No sterols,
 Contains the following organelles: Polysaccharide EXCEPT:
1. Nucleus: bounded by a nuclear membrane, may vary Ergosterol
Peptidoglycan Mycoplasma and
in size, shape, and number. Usually contains one Ureaplasma
nucleolus of mostly RNA. Nucleoid, no
2. Chromosomes: described as linear. Small, bound by
Nucleus nuclear
3. Ribosomes: present at all times for protein nuclear membrane
membrane
synthesis. Chromosome Linear Circular
4. Mitochondrion: generation of energy. Ribosomes 80s 70s
5. Endoplasmic reticulum DAP
6. Vacuoles α-aminodiphate
Lysine synthesis (Diaminopimelate
pathway
pathway)
 IDENTIFICATION OF YEASTS MACROSCOPIC DIFFERENTIATION OF YEASTS AND MOLDS
 Identification relies on a combination of Macroscopic
1. Morphological includes:
a. Color of colonies
b. Size and shape of cells
c. Presence of capsule around the cell
d. Production of hyphae, pseudohyphae,
arthroconidia, and (or) chlamydospores
2. Physiological, and Characteristics:
3. Biochemical characteristics Mold cells Yeast cells
a. Assimilation and fermentation of sugars  Growth can be  As seen in culture, colonies
b. Assimilation of nitrate observed through may appear as white opaque,
 Most commercial tests (eg., ID YST, bioMérieux) are obverse (surface) or pasty to creamy to mucoid
based on sugar assimilation of isolates. reverse (back). (encapsulated) with 0.5 to 3
 DNA sequencing Obverse observation mm in diameter.
 Matrix-assisted laser desorption ionization-time of flight can be white to  Colonies grow at 35-37°C,
mass spectrometry (MALDI-TOF) cream, bright or optimum
light gray to brown,
IDENTIFICATION OF MOLDS reverse may be non-
 Most can be identified after growth in culture. pigmented, to
Recognizable criteria for recognition often differ from the yellow, orange, or
fundamental characteristics as the basic of classification. red
1. Macroscopic characteristics viz. colonial form,
surface color (obverse) pigmentation, reverse CLASSIFICATION OF HYPHAE ACCORDING TO THE PRESENCE
pigmentation, and growth rate. OF SEPTUM
 Factors that influence the colonial form: Septate  Septate hyphae are
1. Culture medium fungus with cross walls.
2. Incubation temperature  From the illustration
3. Age of culture (left), septate hyphae
4. Amount of inoculum that are nearly parallel to
 Sporulation is important in speciation, hence it is one another with
important to select culture conditions which favor dichotomous branching
sporulation. at acute angles —
 Sabouraud’s dextrose agar (SDA) promotes over Aspergillus spp.
production of mycelium which results in loss of Aseptate  From the illustration
sporulation, in such cases, subculture to a low-nutrient (left), a Mucormycosis
medium (stimulate sporulation). with pauciseptate
(coenocytic) with non-
MICROSCOPIC DIFFERENTIATION OF YEASTS AND MOLDS parallel sides.
Microscopic  Aseptate are fungus
without cross walls.
Coenocytic hyphae
results from nuclear
division within a cell
without division in the
Mold Yeast cytoplasm.

Characteristics:
Mold cells Yeast cells
 Compose of the  Cells are mostly spherical to
basic unit of fungus ellipsoidal of 3 to 15 microns
(hyphae). in size. Multiply via asexual
 The septum, reproduction known as
represents the cross budding.
walls (divisions) of a  Stages of budding:
hyphae. 1. Bud emergence
 Mass of hyphae will 2. Protuberance
produce mycelium. 3. Elongation
CLASSIFICATION OF HYPHAE ACCORDING TO MORPHOLOGY Hyaline (ie., Saksenaea  Clear, transparent, and
Pectinate (eg., Microsporum  Multiple projections in a vasiformis — agent of colorless hyphae.
audouinii) hypha resembling an old mucormycosis)
comb hyphae
appearance.

Dematiaceous  Hyphae having structures

that are brown to black,
this is due to the
Racquet (eg., Coccidioides  Hypha with club-shaped
melanotic pigment in the
spp.) cells, the layer end of
cell walls.
one cell being attached
to the smaller end of an
adjacent.

APICAL ELONGATION

Spiral  Hypha forming coiled or

corkscrew like turns.
 Characteristics of
Trichophyton
mentagrophytes.

1 Hyphal wall 7 Nucleus

2 Septum 8 Endoplasmic reticulum
Favic chandelier  Terminal hyphae
3 Mitochondrion 9 Lipid body
branches that are
irregular, broad, and 4 Vacuole 10 Plasma membrane
antler-like in 5 Ergosterol crystal 11 Spitzenkörper
appearance. Especially 6 ribosome 12 Golgi apparatus
characteristics of T.
schöenleinii. Lateral Elongation
Nodular  Round knot-like  Triggered by stems from the sub apical accumulation of
structure formed by wall precursors (presumable vesicles) reaching a critical
intertwined hyphae; concentrations.
seen especially among
dermatophytes. Process of Apical Elongation
I. Cell vesicles move towards the tip of the hyphae
(towards the plasma membrane) where they release
various enzymes and other compounds.
II. Enzymes release by these particles involved in the lysis
and synthesis of the cell wall
III. Following lysis of the cell wall, cell division and growth
Rhizoids  Root like structure along allows for the hyphae to be elongated at the tip.
the vegetative hyphae. IV. Once the new cell has formed, enzymes at the tip start
 Characteristics of certain synthesizing a new cell wall around the new cell to
Glomeromycota protect it. Followed by strengthening of the actin cap.
(formerly Zygomycota) Then disappearance of Spitzenkörper.
(Rhizopus and Absidium). V. Spitzenkörper, play an important role in organizing
(regulating) hyphal growth. Found behind hyphal tip
(apex).
 CLASSIFICATION OF HYPHAE ACCORDING TO HYPHAL  Asexual reproduction
GROWTH o Sporulation and spores are preferred terms used
 Apical elongation leads to the formation of filamentous when there is a merging of nuclear material or
(fuzzy) colonies — mycelium genes combined.
 Types of mycelium: o Asexual spores — spore formed by budding, no
1. Vegetative: buried down, for the water exchange fusion of nuclei takes place in the formation of
and nutrient absorption. spore.
2. Aerial mycelium: projects on the surface of the i. Unicellular
culture medium with reproductive structures o Do not form mycelium, produce
known as spores which can be sexual or asexual. pasty type of colony (yeast).
ii. Multicellular
FUNGAL REPRODUCTION o Form mycelium, produce
 Sexual reproduction filamentous colony (mold).
o Requires the formation of special clusters for
fertilization and nuclear fission. TYPES OF ASEXUAL REPRODUCTION
o Gametangium — sex organ, gametes (sex cells) and Sporangiospores  Spores contained in
either monoecious or dioecious. sporangia or sacs are
o Involves meiosis produced terminally (at
o Sexual spores are developed as a result of a primary the end).
nuclear fusion with reduction in chromosome  Sporangiophore
number during their formation. (specialized stalk which
o Germinates and forms hyphae and mycelium. bears the sporangia).
 Observed among,
TYPES OF SEXUAL REPRODUCTION Zygomycetes, Rhizopus.
Ascospores (eg., Saccharomyces  2-8 ascospores within
Mitospores (Conidiospores)  Results from the
spp., Ascomycetes) an ascus (sac), the
transformation of a
shape of each aid in
vegetative yeast or
identification of
specialized hypha called
fungus.
conidiophores. Often
referred to as macro-
(multicellular) and
microconidia
Basidiospores (eg.,  2-4 basidiospores on (unicellular).
Basidiomycetes the surface of a club-  Freed by abstraction at
shaped called the point of attachment.
basidium.  Observed among
Ascomycetes and
Deuteromycetes.
Thallospores — produced on a thallus
Blastospores (eg., Candida  Asexual spore formed
Zygospores (eg., Mucor &  When two spp.) from a budding process
Rhizopus) sporangiophores or blowing out along the
sexually fuse to form mycelium or from
a large thick-walled another blastospore —
bodies. blastogenesis
 Zygosporangium  Blastoconidia
contains zygospores
along a nonseptate Arthrospores (eg.,  Formed by
hyphae. Dermatophytes, Coccidioides fragmentation
immitis) (segmentation) or
Oospores (eg., Phythophthora,  Produced from a
phythium, etc.) fusion of two (2) non- disarticulation of hyphae
identical hyphae. (mycelium) which results
to cutting of rectangular
thickwalled spore.
Chlamydospores (eg.,  Formed by budding from
Candida spp.) a pseudohypha
 Thick-walled resting
spores either within
(intercallary) the
segments; hyphal sides
(sessile); or hyphal tip
(terminal).

CLASSIFICATION OF HYPHAE ACCORDING TO THE PRESENCE

OF SEPTUM
Phialospores — highly developed conidiophores
Phialospores (eg.,  Vesicle, enlarged swollen
Aspergillus spp., Penicillium cell often at the end of a
spp.) conidiophore or
sporangiophore and it
bears conidia.
 The conidiospores are
formed usually at the
sides (tips) of hyphae
and are usually in chains.

Site of
Classification Description Examples
infection
Do not
Outer skin Malassezia
Superficial penetrate
layer (hair) furfur
tissues
Do not Trichophyton
Skin, hair,
penetrate species
nails
Cutaneous tissues, but
(keratinize
can cause Microsporum
d area)
inflammation species
Sporothrix
Penetrate schenkii
Connective
tissues,
tissues,
Subcutaneous transmitted Rhinosporidi
Muscles,
via traumatic um seeberi
Bones
inoculation
Loboa loboi
All body Thermal Histoplasma
Systemic
system (dimorphic capsulatum
Aspergillus
Infects
fumigatus
All body immunocom
Opportunistic
system promised
Candida
host
albicans