Microbiology (Dr.

De Castro)
Introduction to Mycology

13 December 07

INTRODUCTION TO MYCOLOGY • Poisoning of man and animals by feeds and food

products contaminated by fungi which produce toxins
Mycology from the grain substrate
• The study of fungi and their multiple functions in nature.
3. Mycetismus
FUNGUS • The ingestion of pre-formed toxin mushroom poisoning
4. Infection
• Eukaryotic – a true nucleus
B. MORPHOLOGY OF FUNGI
• Do not contain chlorophyll  achlorphyllous • Yeast
• Have cell walls • Yeast-like
• Produce filamentous structures • Mold (hyphae)
• Produce spores • Dimorphic Fungi
• Grow as saprophytes and decompose dead matters
• 100,000-200,000 species YEASTS
• 300 species are pathogenic in man 1. Unicellular organisms
2. Round or oval fungi that bud or pinch off
FUNGAL CELL WALL COMPOSITION 3. Growth resembles bacteria
− Structural Components 4. May be dry, pasty or slimy
o Chitin microfibrils [β (1-4)-linked polymer of N- 5. May be gram-positive
acetylglucosamine] 6. Much larger than bacteria
o Chitosan in Zygomycota [β (1-4)-linked polymer of • Bacteria are 0.1-0.2 microns
glucosamine] • Yeast are 3-20 microns (the size of PMNs, RBCs, etc
o Β-linked glucans Ex. C. neoformans
− Gel-Like Components
o Mannoproteins (form matrix throughout wall) YEAST-LIKE (PSEUDOHYPHAE) – Candida
− Antigenic glycoproteins, agglutinans, adhesions • Budding yeast with septum. The septum has formed
− on cell wall surface between the daughter bud and the mother cell but
− Melanins separation of the two has not occurred.
− dark brown to black pigments • Candida albicans mother and daughter cells. Cells were
− confer resistance to enzyme lysis, confer grown under conditions that induced hypha formation for 30
mechanical strength and protect cell from UV light, mins. The daughter cell is on the right; the mother cell is on
solar radiation and dessication the left. The daughter cell has not reached a threshold
− Sporopollenin volume and therefore has not yet formed a hypha.
− Aromatic polymer found in spore walls of some • C. albicans cell at 3h. 3 hours after the appearance of the
fungi, confers properties similar to melanin germ tube, the hypha has septa.
− Plasma membrane
• C. albicans hyphal cells at 5h. After 5h in hypha-inducing
− Semi-permeable
medium, many hyphae are evident. Clumping of the hyphae
is also apparent and hyphae are beginning to form hyphae
A. CLASSIFICATION
blastospores, which are now budding cells.
TAXONOMY
HYPHAE (MYCELIUM)
Kingdom Characteristic Examples
• Multi-cellular filamentous structures
Monera Prokaryocyte Bacteria
• Cylindrical, branching filaments composed of tubular cell
Actinomyces
wall filled with cytoplasm and organelles
Protista Eukaryocyte Protozoa
• Most fungal hyphae are 2-10 µm diameter
Fungi Eukaryocyte Fungi
Plants Eukaryocyte Plants • A mass of hypahe is called Mycelia
Moss • Pattern of branching of mycelia aids diagnosis
Animals Eukaryocyte Anthropods • “hypahe”, “mycelium”, “molds” – interchangeable
Mammals
Man Molds (Hyphal Fungi)
1. Seen microscopically as filaments which are variable in
MEDICAL MYCOLOGY length but usually constant in diameter
There are four types of mycotic diseases: 2. On plates look fuzzy or “carpet-like”
1. Hypersensitivity
− An allergic reaction to molds and spores Hyphae
− Indoor pollution 1. Septate – filaments have crosswalls or septate
Ex. Aspegillus
2. Mycotoxicoses
2. Aseptate – filaments lack crosswalls; also called coenocytic
Ex. Mucor

Virns, leu, brim 1 of 4

 Microbiology – Intro to Mycology by Dra De Castro Page 2 of 4

3. Vegetative – growth within the surface of the medium (agar)

4. Aerial – filaments that extend above the surface (stick up HOW DO PEOPLE GET MYCOSES?
into the air) ● Overgrowth of normal flora (usually candida) after
catheters, chemotherapy or broad-spectrum antibiotics
Septa ● Inhalation of conidia (Histoplasmosis)
• Regular cross-walls formed in hyphae. Hyphae with septa ● Trauma / Implantation (Sporothrix)
● Contact with plants (Sporothrix) and animals
are septate, those lacking septa except to delimit
(dermatophytes)
reproductive structures and aging hyphae are called
aseptate or coenocytic.
ESTABLISHMENT OF INFECTION WITH MYCOTIC AGENT
o Primary septa are formed as a process of hyphal
DEPENDS ON:
extension and generally have a septal pore, which
● Inoculum size
allows for cytoplasmicand organelle movement.
● Resistance of the host
o Secondary or adventitious septa are imperforate,
formed to wall off ageing parts of the mycelium. SEVERITY OF DISEASE
Septate hyphae- Aspergillus ; Aseptate hyphae - Mucor  In mycotic infections depends more on the host immune
DIMORPHIC FUNGI (FUNGAL DIMORPHISM) system than on the virulence of the fungus
Yeast
 Clinician must distinguish bet :
(Parasitic or Pathogenic Form)
● Colonization
• Seen in human tissue, exudates, or if cultured in an
● Transient fungemia
incubator at 37 °C. ● True infection
Mycelium
(Saprophytic or Mold Form) PORTAL OF ENTRY
• Form observed in nature or if incubated at 25 °C. ● Skin
• Conversion to the yeast form essential for pathogenicity ● Hair
● Nails
C. REPRODUCTION OF FUNGI ● Respiratory tract
SEXUAL REPRODUCTION ● Gastrointestinal tract
● Used for scientific classification ● Urinary tract
● Rarely seen in tissue
● Not usually studied in the laboratory COLONIZATION
● Probably not infectious ● Multiplication of an organism at a given site without harm to
the host
ASEXUAL REPRODUCTION
● These are the forms we study in the laboratory INFECTION
● Asexual reproductive forms are CONIDIA ● Invasion and multiplication of organisms in body tissue
● Sometimes called SPORES resulting in local cellular injury
● Usually round but may be rectangular
● Similar in size to yeasts D. DIAGNOSIS
SPECIAL STAINS FOR FUNGI
TYPES OF CONIDIA ● Giemsa stain
1. Arthroconidia o Nucleus blue with clear or pink cytoplasm
o Hyphae break or fragment at separation points forming ● Gomori’s Methenamine Silver (GMS) / Silver stain
small pieces that give rise to a new hyphae o Yeasts and hyphae are brown-black, the counterstain is
o E.g. Coccidioides, Geotrichum usually green, stains the cell wall only
2. Blastoconidia ● Periodic Acid Schiff (PAS)
o Cells reproduce by budding o Stains cell walls pink
o Daughter cells pinch off from the mother cell o Counterstain is blue or green
o E.g. Canidia, Cryptococcus I. Wet Mount
o Note: If blastoconidia continue to elongate and fail to II. Skin Test
pinch off, they are called PSEUDOHYPHAE (yeast-like) III. Serology
3. Chlamydoconidia IV. Fluorescent antibody
o Round and thick-walled conidia that form directly within V. Biopsy and Histopathology
or at the ends of hyphae; they can then fragment or VI. Culture
brek off to form new hyphae DIRECT MICROSCOPIC OBSERVATION
o E.g. Candida albicans, dermatophytes ● 10% KOH
● Skin crappings suspected to contain dermatophyets or pus
OTHER PROPERTIES OF FUNGI from a lesion can be mounted in KOH on a slide and
● They resemble mammalian cells examined directly under the microscope
● Cell wall has chitin glucosamine, hexose, hexosamine, ● Gentle heat
protein, etc. SKIN TESTING (DERMAL HYPERSENSITIVITY)
● Special stains may be required to see them ● May interfere with serologic test, causing false positive
result
WHY ARE WE SEEING MORE MYCOSES TODAY? ● Use is limited to:
● More compromised patients o Determine cellular defense mechanisms
● Transplant patients o Epidemiologic studies
● Anti-cancer drugs SEROLOGY
● Invasive catheters (bladder, CNS, vascular) ● May be helpful when it is applied to a specific fungal disease
● Use of broad-spectrum antibiotics
● The host has changed, not the fungi
 Microbiology – Intro to Mycology by Dra De Castro Page 3 of 4

● There are no screening antigens for fungi in general –fungi

are poor antigens
Squalene
● The efficacy of serology varies with different fungal
Allylamines
infections
Squalene-2,3-Epoxide
FUNGAL SEROLOGY
ANTIBODIES
LATEX AGGLUTINATION IgM Lanosterol
IMMUNO-DIFFUSION IgG Azoles
COMPLEMENT FIXATION IgG Morpholines
Most serology tests for fungi measure antibody. Newer tests to Polyenes
measure antigen are now being developed.
Ergosterol
ANTIGEN DETECTION PRESENTLY AVAILABLE
● Cryptococcosis
● Histoplasmosis
PRIMARY ANTIFUNGALS
DIRECT FLUORESCENT ANTIBODY
1. Polyene derivatives
● Can be applied to:
 Amphotericin B
o Tissue
MOA:
o Culture
 Binds to sterols
BIOPSY AND HISTOPATHOLOGY  Ergosterol is a constituent of the cell walls of the
● Normal host fungi (eukaryotic)
o Pyogenic  There is a greater avidity for ergosterol than for
o Granulomatous cholesterol in the cell walls of animal and human
● Immunodeficient host cell
o Necrosis  The binding to the cell wall alters the permeability
CULTURE of the cell wall and results in the leakage of
● A definitive diagnosis requires a culture and identification intracellular contents of the fungus
● Pathogenic fungi are usually grown on SABOURAUD  Nystatin
DEXTROSE AGAR 2. Azoles (Ketoconazole, Flucanozole, Itraconazole,
● Isolation media Voriconazole)
o Sabouraud Dextrose Agar (pH is approximately 5.6)  There are few rare serious side effects from
o Plain each itraconazole and flucanozole
o With antibiotics 3. Griseofulvin
o With cycloheximide  A slow acting drug used for skin and nail infections.
● Incubation temperature It accumulates in the stratum corneum and prevent
o 37°C: body temperature hyphal penetration through these layers
o 25°C: room temperature 4. Allylamines
 Terbinafine (Lamisil)
o Two cultures are inoculated and incubated separately at 5. 5- fluorcytosine
25°C and 37°C to reveal DIMORPHISM  Interferes with RNA synthesis
o The culture are examined macroscopically and 6. Echinocandins
microscopically  Caspofungin
o They are not considered negative for growth until 4
weeks of incubation MOA
Polyenes Ergosterol in cell membrane
E. TREATMENT Azoles Interfere with ergosterol synthesis
THERAPY Griseofluvin Forms a barrier to fungal growth
Since they are eukaryotic, fungi are biochemically similar to 5-FC Inhibits RNA syntheis
the human host. Thus, it is difficult to develop
chemotherapeutic agents that will destroy the pathogenic F. CLINICAL CLASSIFICATION
fungus and not harm the patient  Cutaneous
A BASIC TENET OF PATHOLOGY  Subcutaneous
A cause of irreversible cell injury is cell membrane damage  Systemic
IN FUNGAL THERAPY  Opportunistic
We attempt to induce cell injury by causing the cell
membrane of the fungus to become permeable Superficial Mycoses
PROBLEM  Skin, hair and nails
Finding an agent that will selectively injure fungal cells  Rarely invade deeper tissue
without damaging host cells  Dermatophytes
ALL EUKARYOTIC CELLS CONTAIN Subcutaneous Mycoses
Mammalian Cells
 Confined to subcutaneous tissue and rarely spread
 Cholesterol systemically. The causative agents are soil
Fungal Cells organisms introduced into the extremities by
 Ergosterol trauma
Systemic Mycoses
Ergosterol Synthesis  Involve skin and deep viscera
Acetyl-CoA
Acetoacetyl-CoA  May become widely disseminated
HMG-CoA  Predilection for specific organs
Mevalonic Acid Opportunistic Fungi
 Microbiology – Intro to Mycology by Dra De Castro Page 4 of 4

 Ubiquitous saprophytes and occasional pathogens

that invade the tissues of those which have:
o Predisposing diseases: diabetes, cancer,
leukemia, etc
o Predisposing conditions: agammaglobinemia,
steroid and antibiotic therapy

Yung 1st week trans bout myco.. antay antayin nio lang ha.. di pa
nman kelngan. Or kelangan nio pa ba? O well, bsta mgpapass
kami.. late late konti. Kasi kakabigay lng ng mga copies.. peace!..
doble doble na trans na ggwins.. whaaa!.. No trans days are so
over.. hay.. kakaiyak.malas namin lately..