Introduction To Medical Mycology

Mycology MD 3110 Semester 2
Introduction to Mycology
By
Prof. Dr. Afreen Banu
Lecture out line
I- What is Fungi??
⮚ Is any member of the group of eukaryotic organisms that includes
unicellular microorganisms such as yeasts and molds, as well as
multicellular fungi that produce familiar fruiting forms known as
mushrooms.
⮚ These organisms are classified as a kingdom, Fungi, which is

separate from the other life kingdoms of plants, animals, protists,
and bacteria.
Mycology Semester1
1- General Characters
⮚ Fungi studies are called mycology
⮚ Most fungi like molds & mushrooms are
multicellular, but yeast are unicellular
Mycology MD3111 Semester 2
1- General Characters
o Filaments of fungi are called hyphae.
o The cell walls contain chitin.
o The MYCELIUM is a mat of hyphae visible to
the unaided eye ( bread mold)
o Some hyphae may divided by cross sections
called septa
2- Sexual Reproduction
3- Asexual Reproduction
Vegetative cell
Mycology Semester 2
Reproduction
❖When environmental conditions are favorable,

asexual reproduction occurs rapidly.
❖When unfavorable conditions stress the

organism, sexual reproduction occurs, and the
offspring have an increased like hood that they
will be better suited for the environment.
II- Classification and Properties of Fungi
Depend up on Characterization
Phylum Common Name Characteristics Examples

Zygomycota Bread molds Display conjugation Rhizopus & other
bread molds
Ascomycota Sac fungi Produce asci & Neurospora,

ascospores during Penicillium,
sexual reproduction Saccharomyces, and
other yeast
Basidiomycota Club fungi Produce basidia & Amanita & other

basidiospores mushrooms
Deuteromycota Fungi Imperfecti Sexual stage Soil organisms,
nonexistent or various human
unknown, hence pathogens
“imperfect”
1- Deuteromycota
DEUTEROMYCETES
“Fungi imperfecti”
• Fungi that DO NOT or NOT KNOWN produce sexual

spore.
• Production of septate mycelium and/or yeasts
• Fungi producing the antibiotic penicillin and those that
cause athlete's foot and yeast infections are imperfect
fungi
2 ZYGOMYCOTA
FUNGAL CLASSIFICATION: ZYGOMYCOTA
• Mostly aseptate
• Asexual spores: sporangiospore
• Sexual reproduction: zygospore
3 BASIDIOMYCOTA
FUNGAL CLASSIFICATION: BASIDIOMYCOTA
“Club Fungi”
• Hyphae septate (hyphae are dikaryotic)

– Easily distinguish by presence of clamp connections over the
septa, spores produced externally
• Sexual reproduction: Basidiospores
4 ASCOMYCOTA
FUNGAL CLASSIFICATION: ASCOMYCOTA
• Hyphae septate, fungi produced

within a sac called an ascus.
• Asexual reproduction: Conidia

• Sexual reproduction: Ascospores
(Eight in ascus)
Importance of Fungi
✔ Fungi decompose dead plant and animal matter.
✔ Fungi in Humans-causes allergy.
✔ Fungi in industry.
✔ Fungi as Plant pathogens.
✔ Fungi as Human and Animal Pathogens
III- Human diseases caused by Fungus
⮚ Fungal diseases (mycoses) are usually divided in 5 groups according
to the level of infected tissue & mode of entry into the host.
Fungal diseases Infected area
Superficial mycoses Occurs mainly in the tropics
& include Black piedra,
White piedra, Tinea versicolor
1
Cutaneous mycoses Those of the outer layer of the skin, generally called
ringworms, tineas or dermatomycoses
2
Subcutaneous mycoses Below the skin,
example chromomycosis
3
Systemic mycoses Acquired by the inhalation of spores from soil in
which the mold phase of the fungus resides-
4 Histoplasmosis
Opportunistic mycoses Candidiadis-Mycosis caused by dimorphic fungus,

Candida albicans
5
IV- Laboratory Diagnosis
1. Direct Microscopic examination:
❖Skin scrapings, Sputum, lung biopsy.
❖Mount it with KOH (Potassium hydroxide)
2. Culture of the organism in media.
3. Serologic test:
❖Test for the presence of antibodies in patient
serum/spinal fluid.
4. DNA probe test by PCR.
❖Can identify colonies growing at early stage
Microscopic Morphology
• Definitive means of
identification
• Evaluate:
– Shape
– Method of production
– Arrangement of
conidia/spores
– Size and color of hyphae
(1) Specimen Collection
• Specimen types.
• Collect from area most likely infected.
• Use sterile technique.
• Keep specimen moist.
• Label container properly.
• Transport right away.
• Process right away.
(2) Direct Examination
• Provides preliminary report
• Guides MD in treatment of patient
• Observe yeast phase of dimorphic
• Gives clues to its causative agent
• Inoculate special media
• May require more than one direct
examination method
(3)- Staining and Direct Examination
• Saline wet mount.
• Lactophenol Cotton Blue (LPCB) wet mount.
• 10% KOH preparation.
• Gram stain.
• Acid fast stain.
• India ink stain.
Lactophenol Cotton Blue (LPCB)
Principle
• Phenol: kills any live organisms;
• Lactic acid : It preserves fungal structures
• Cotton blue : It stains the chitin in the
fungal
cell walls.
Direct Examination
• Calcofluor white stain (CWS).
• Gomori Methenamine Silver stain (GMSS).

(4) Specimen Processing
• Safety:
⮚Tube media preferred over
plate media.
⮚Work in safety hood.
⮚Wear gloves and lab coat.
⮚Autoclave specimens and
media.
⮚Disinfect work area daily.
(5)-Fungal Culture Process
1. Specimen collection and transportation
2. Direct examination of specimen
3. Selection and inoculation of media
4. Evaluation of fungal growth
5. Serological testing
6. Antifungal susceptibility testing
(6) Media for fungal culture
1. Sabouraud Dextrose Agar (SDA):
2. Potato Dextrose Agar (PDA):
3. Brain Heart Infusion (BHI) with/without 5%

blood and 1% glucose:
Subculture / Identification Media
A. Neutral Sabouraud
Dextrose Agar: (a type of
agar containing peptone for
fungi growth)-SDA
B. Potato Dextrose Agar (PDA):
C. Cornmeal-Tween 80 agar:
D. Niger Seed Agar (Bird Seed
Agar):
E. Tween 80 / Oxgall / caffeic
acid agar:
(pH=5.5-6.5 at 25-30 ºC)
Examination of Culture Growth
1- Potential pathogens:
⮚ Slow growers.
⮚ Growth on Mycosel.
⮚ Color: dull buff, brown and
mousy gray.
⮚ Dimorphic.
2- Growth rate:
⮚ Rapid growers: 1-5 days.
⮚ Intermediate growers: 6-10 days.
⮚ Slow growers: >10 days.
(7)- Serological Diagnosis
⮚ Immunodiffusion: (a technique for detecting or
measuring antibodies and antigens by their
precipitation when diffused together through a
gel or other medium).
⮚ EIA (Enzyme immunoassay).
⮚ Latex agglutination.
V- Antifungal Susceptibility
1- Determine appropriateness:
⮚ Standardization of testing
(Susceptibility or MIC).
⮚ Methods.
⮚ Predictability in vitro and vivo.
2- Antifungal Agents for Treatments:
⮚ Antifungal Susceptibility Test and
MIC: with active antibiotic with no
effect host (Eukaryotic).
⮚ Antifungal antibiotics targets: Cell
Wall and Cell Membrane of Fungi
(Chitin and Ergosterol), only few
antibiotics active from more than 50.
VI)- References
Main Reference:
1. Michael Ford 2015. Medical Microbiology. 2nd revised edition. Oxford University Press.
Oxford, United Kingdom.
2. Jawetz, Melnick and Adelberg's 2015. Medical Microbiology, 27th Edition. LANGE.
3. Levinson, Chin-Hong, Joyce, Nussbaum and Schwartz 2015. Review of Medical
Microbiology and Immunology. Fifteenth Edition. McGraw Hill Education.
4. Karin C. Van Meter and Robert J. Hubert 2016. Microbiology for the Healthcare
Professional. 2nd Revised edition. Elsevier - Health Sciences Division.
5. Patrick Murray, Ken Rosenthal and Michael Pfaller 2016. Medical Microbiology. 8th
Edition. Elsevier.
6. Luis Miguel, de Pablos Torró and Jacob-Lorenzo Morales 2018. Protozoan Parasitism:
From Omics to Prevention and Control. Caister Academic Press.
Additional Reference:
7. Jacquelyn G. Black 2014. Microbiology Principles and Explorations. 9th Edition. Wiley-
Blackwell.
8. Richard A Harvey 2016. Specifications of Lippincott Illustrated Reviews Microbiology, 3rd
Edition SAE. Williams and Wilkins.
9. Karen C. Carroll, Stephen A. Morse, Timothy A. Mietzner and Steve Miller (2016).
Medical Microbiology. 27th Edition. Mc Grow Hill-Lange.
Web links:
⮚http://www.microbiologybook.org/fox/protype.htm
⮚http://www.microbiologyinpictures.com/neisseria%20meningitidis.html
⮚https://www.boundless.com/microbiology/textbooks/boundless-microbiology-textbook/cell-
structure-of-bacteria-archaea-and-eukaryotes.
THANK YOU

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Mycology MD 3110 Semester 2

⮚ These organisms are classified as a kingdom, Fungi, which is

❖When environmental conditions are favorable,

❖When unfavorable conditions stress the

Phylum Common Name Characteristics Examples

Ascomycota Sac fungi Produce asci & Neurospora,

Basidiomycota Club fungi Produce basidia & Amanita & other

• Fungi that DO NOT or NOT KNOWN produce sexual

FUNGAL CLASSIFICATION: ZYGOMYCOTA

• Hyphae septate (hyphae are dikaryotic)

• Hyphae septate, fungi produced

• Asexual reproduction: Conidia

Opportunistic mycoses Candidiadis-Mycosis caused by dimorphic fungus,

• Gomori Methenamine Silver stain (GMSS).

3. Brain Heart Infusion (BHI) with/without 5%

⮚ EIA (Enzyme immunoassay).

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