Nina Ian John “G” Rachel Mark Jocelle Edo Gienah Jho Kath Aynz Je Glad Nickie Ricobear

Teacher Dadang Niňa Arlene Vivs Paul F. Rico F. Ren Mai Revs Mavis Jepay Yana Mayi Serge Hung Tope Ag Bien

S3 L22: Introduction to Mycology by Dra. Madrid DDeecceem

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MYCOLOGY FUNGAL MORPHOLOGY

is the study of fungi and their multiple functions in nature
yeast
FUNGUS yeast-like
eukaryotic – a true nucleus mold (hyphae)
do not contain chlorophyll dimorphic fungi
have cell walls
produce filamentous structures Yeasts
produce spores
unicellular organism
FUNGAL CELL WALL COMPOSITION round or oval fungi that bud or pinch off
growth resembles bacteria
 Structural Components may be dry, party, or slimy
chitin microfibrils [β(1-4)-linked polymer of N-acetylglucosamine] may be gram-positive
chitosan in Zygomycota [β(1-4)-linked polymer of glucosamine] much larger than bacteria – bacteria are 0.1-0.2 microns; yeasts are 3-20
β-linked glucans microns (the size of PMNs, RBCs, etc.)
 Gel-like Components
mannoproteins (form matrix throughout wall) Hyphae (Mycelium)

Other Cell Wall Components multicellular filamentous structures

antigenic glycoproteins, agglutinans, adhesions – on cell wall surface cylindrical, branching filaments composed of a tubular cell wall filled with
melanins – dark brown to black pigments (confer resistance to enzyme cytoplasm and organelles
lysis, confer mechanical strength and protect cells from UV light, solar most fungal hyphae are 2-10 µm diameter
radiation, and desiccation) a mass of hyphae is called mycelia
sporopollenin – aromatic polymer found in spore walls of some fungi pattern of branching of mycelia aids diagnosis
(confers properties similar to melanin) “hyphae”, “mycelium”, “molds” – interchangeable
plasma membrane – semi-permeable
Molds (Hyphal Fungi)
TAXONOMY
seen microscopically as filaments which are variable in length but usually
Kingdom Characteristic Example constant in diameter
Monera Prokaryocyte Bacteria
Actinomyces Hyphae
Protista Eukaryocyte Protozoa 1. septate – filaments have crosswalls or septae
Fungi Eukaryocyte Fungi 2. aseptate – filaments lack crosswalls; also called coenocytic
Plants Eukaryocyte Plants 3. vegetative – growth within the surface of the medium (agar)
Moss 4. aerial filaments that extend above the surface (stick up into the air)
Animals Eukaryocyte Arthropods
Mammals Septa
Man
Regular crosswalls formed in hyphae
Hyphae with septa are septate, those lacking septa Except to delimit
MEDICAL MYCOLOGY reproductive structures and aging hyphae are called aseptate or
coenocytic
Four types of mycotic diseases:
Primary septa are formed as a process of hyphal extension and generally
have a septal pore, which allows for cytoplasmic and organelle movement
1. Hypersensitivity
Secondary or adventitious septa are imperforate, formed to wall off
an allergic reaction to molds and spores
ageing parts of the mycelium
indoor air pollution
Dimorphic Fungi (Fungal Dimorphism)
2. Mycotoxicoses
poisoning of man and animals by feeds and food products  Yeast (parasitic or pathogenic form)
contaminated by fungi which produce toxins from the grain seen in human tissue, exudates, or if cultured in an incubator at
substrate 37OC
 Mycelium (saprophytic or mold form)
3. Mycetismus
form observed in nature, or if incubated at 25OC
the ingestion of preformed toxin
conversion to the yeast form essential for pathogenicity
mushroom poisoning
FUNGAL REPRODUCTION
4. Infection
 Sexual Reproduction
used for scientific classification

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 rarely seen in tissue respiratory tract
not usually studied in the laboratory gastrointestinal tract
probably not infectious urinary tract

 Asexual Reproduction Colonization

these are the forms we study in the lab Multiplication of an organism at a given site without harm to the host
asexual reproductive forms are conidia
sometimes called spores Infection
usually round but may be rectangular Invasion and multiplication of organisms and body tissues resulting in
similar in size to yeasts local cellular injury (respiratory tract, mouth, eye, skin, urogenital tract,
anus)
Types of Conidia
Special Stains for Fungi
1. blastoconidia
cells reproductive by budding 1. Giemsa stain
daughter cells pinch off from the mother cell nucleus blue with clear or pink cytoplasm
e.g. Candida, Cryptococcus 2. GMS (Gomori’s Methenamine Silver) or Silver Stain
note: if blastoconidia continue to elongate and fail to pinch off, they yeasts and hyphae are brown black
are called pseudohyphae (yeast-like) the counterstain is usually green
stains the cell wall only
2. arthroconidia 3. PAS (Periodic Acid Schiff)
hyphae break or fragment at separation points forming small pieces stains cell walls pink
that give rise to new hypha counterstain is blue or green
e.g. Coccidioides, Ceotrichum
DIAGNOSIS
3. Chlamydoconidia
round and thick-walled conidia that form directly within or at the ends of 1. Wet Mount
hyphae; they can then fragment or break off to form new hyphae 2. Skin test
e.g. Candida albicans, dermatophytes 3. Serology
4. Fluorescent antibody
Other Properties of Fungi 5. Biopsy and histopathology
they resemble mammalian cells 6. Culture
cell wall has chitin glucosamine, hexose, hexosamine, protein etc.
Direct Microscopic Observation
special strains may be required to see them
1. 10% KOH (KOH wet mount) Skin
Why are we seeing more mycoses today?
scrapings suspected to contain
more compromised patients
dermatophytes or pus from a lesion
transplant patients can be mounted in KOH on a slide
anti-cancer drugs and examine directly under the
invasive catheters (bladder, CNS, vascular) microscope.
use of broad-spectrum antibiotics
the host has changed, not the fungi 2. Gentle heat

How do people get mycoses? Skin Testing (Dermal Hypersensitivity)

overgrowth of normal flora (usually Candida) after catheters, May interfere with serologic test causing false positive result.
chemotherapy, or broad spectrum antibiotics Use is limited to:
inhalation of conidia (Histoplasmosis) o Determine cellular defense mechanisms
trauma/implantation (Sporothrix) o Epidemiological studies
contact with plants (Sporothrix) and animals (dermatophytes)
Serology
Establishment of infection w/ a mycotic agent depends on: may be helpful when it is applied to a specific fungal disease
1. inoculum size there are no screening antigens for fungi in general – fungi are poor
2. resistance of the host antigens
the efficacy of serology varies with different fungal infections
Severity of Disease Most serologic tests for fungi measure antibody.
in mycotic infections depend more on the host immune system than on Newer tests to measure antigen are now being developed. Antigen
the virulence of the fungus detection presently available for Cryptococcosis, Histoplasmosis
A clinician must distinguish between: Inflammatory reaction
colonization
transient fungemia 1. Normal host
true infection Pyogenic
Granulomatous
PORTAL OF ENTRY 2. Immunodeficient host
skin Necrosis
hair
nails

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Culture Fluconazole
A definitive diagnosis requires a culture and identification. Itraconazole
Pathogenic fungi are usually grown on Saboraud dextrose agar Voriconazole
(pH ~ 5.6) – isolation media
Plain 3. Griseofulvin - A slow activating drug used for skin and nail infections. It
With antibiotics accumulates in the stratum corneum and prevents hyphal penetration
With cycloheximide through these layers.

Incubation temperature 4. 5-fluorocytosine (5-FC) - interferes with RNA Synthesis

Two cultures are inoculated and incubated separately at 25oC (room
temp) and 37oC (body temp) to reveal dimorphism. 5. Allylamines - Terbinafine (Lamisil)
The cultures are examined macroscopically and microscopically.
They are not considered negative for growth until after 4 weeks of 6. Echinocandins (Caspofungin)
incubation.
Mechanisms of Action
Therapy
Since they are eukaryotic, fungi are biochemically similar to the Polyenes – ergosterol in cell membrane
human host. Thus, it is difficult to develop chemotherapeutic agents Azoles – Interfere with ergosterol synthesis
that will destroy the pathogenic fungus and not harm the patient Griseofulvin – Forms a barrier to fungal growth
A basic tenet of pathology: “A cause of irreversible cell injury is cell 5-FC – Inhibits RNA synthesis
membrane damage.”
In fungal therapy, we attempt to induce cell injury by causing cell Clinical Classification
membrane of the fungus to become permeable.
Problem: Finding an agent that will selectively injure fungal cells 1. Superficial Mycoses
without damaging host cells. Skin, hair and nails
Rarely invade deeper tissues
All eukaryotic cells contain sterols. Dermatophytes
Mammalian cells – cholesterol
Fungal cells – ergosterol 2. Subcutaneous Mycoses
Confined to subcutaneous tissue and rarely spread systemically.
Ergosterol synthesis The causative agents are soil organisms introduced into the
extremities by trauma.
Acetyl-CoA
↓ 3. Systemic Mycoses
Acetoacetyl-CoA Involve skin and deep viscera
↓ May become wildly disseminated
HMG-CoA Predilection for specific organs
↓
Mevalonic Acid 4. Opportunistic Fungi
↓ Ubiquitous saprophytes and occasional pathogens that invade the
Squalene tissues of those who have:
Allylamines ↓ o Predisposing diseases – diabetes, cancer, leukemia, etc.
Squalene-2, 3-epoxide o Predisposing conditions – agammaglobulinemia, steroid or
↓ antibiotic therapy
Lanosterol
Azoles
Morpholines
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Polyenes
Ergosterol

Primary Antifungal Agents

1. Polyene Derivatives
A. Amphotericin B
Mechanism of action:
Amphotericin B binds to sterols
Ergosterol is a constituent of the cell walls of fungi (they are
eukaryotic).
There is a greater avidity for ergosterol than for the cholesterol in
the cell walls of animal and human cell.
The binding to cell walls alters the permeability of the cell wall and
results in leakage of the intracellular contents of the fungus.

B. Nystatin

2. Azoles - there are a few rare serious side effects from Itraconazole and
Fluconazole.
Ketoconazole

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