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Bacteriology
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Microbiology
Mycology Helminthology
Protozoalogy
Parasitology
The study of fungi causing
diseases in humans
Objectives
• To impart sufficient basic science of the medically important
fungi to assist you in diagnosing mycotic diseases
• To characterize the following different types of mycoses,
defining the tissue they affect
Superficial
Cutaneous
Subcutaneous
Systemic
Opportunistic
• To differentiate the etiologic agents of these mycosis
• To impart sufficient clinical knowledge
• To raise your index of suspicion for mycotic diseases
• Treatment
Medical Mycology
• Fungi were discovered before bacteria & viruses
• Most fungi cause skin or cosmetic infections
while bacteria & viruses cause fatal diseases
• Clinical Mycology has entered “Golden Age” in
modern medicine due to:
Lacks Contains
• Peptidoglycan • Peptidomannan
• Techoic acids • Glycan (target for
• Lipopolysaccharide new antifungal agents)
Reproductive
Hyphae & conidia
Mycelium
Aerial
(thallus)
hyphae
Vegetative
Surface of hyphae
media
Fungi Groups
On the basis of morphology
2. Yeasts
• Unicellular (rounded or oval)
• Reproduce by budding
• The only example of pathogenic yeast
is Crptococcus neoformans
Fungi Groups
Ascus
Male
Female cell
cell
1. Zygospore 2. Ascospore
Two identical cells Fusion of nuclei of two
form the zygote 4. Oospore
cells in an ascus (sac)
Female cell fertilized
by male cell
Fungi Groups
2. Arthrospores Cubical
Formed within lumen of hyphae Rounded
Size less than the size of hyphae
Can be cubical or rounded
3. Chlamydospores
Produced by swelling from thallus
Fungi : Reproduction
Asexual Spores Spores
disseminate
B. Sporangiospores
Produce sacs filled with spores called sporangium
Hyphae that carry sporangium are called sporangiophores
1. Microconidia
Are unicellular
Fungi : Reproduction
Asexual Spores
C. Exogenous Spores
2. Macroconidia
Are multicellular
Have different shapes
Specimens
Method
• Place the specimen on glass slide
• Add a drop of KOH (20%)
• Place a cover slip
• Gentle heating for 5-10 min (indirect heat)
• Examine under x40 objective
• See the fungal elements : hyphae,
microconidia, macrocondia
Fungi : Lab Diagnosis
I. Direct microscopic Examination
B. Stained Preparation
• A rapid, easy & cheap method
1. Lactophenol cotton blue (LPCB) stain
Place a drop of alcohol on slide
Immerse the specimen in it
Add 1-2 drops of LPCB
Place a coverslip and see under microscope
• Used to see fungal elements in
dermatophyte cultures
Fungi : Lab Diagnosis
I. Direct microscopic Examination
B. Stained Preparation
2. Gram-staining
• For yeasts : stain gram-positive, are much
larger than bacteria
Candida albicans
in Sputum-Gram
Candida albicans-EM
Microsporum
LPCB
Aspergillus
LPCB
Penicillium-EM
Fungi : Lab Diagnosis
c) Biochemical reactions
• like sugar fermentation
• Commercial kits are available
Germ tube test
For identification of Candida albicans
• Place yeasts in serum and incubate at
37oC for 3 hrs
• Most C. albicans will produce tube-like
projections called germ tubes
• Seen under microscope
Fungi : Lab Diagnosis
III. Slide Culture
Is used to:
• See whole morphological details of
fungus especially yeast
• Prevent disturbing the fungal
morphology
• Hyphae & spores remain intact
Fungi : Lab Diagnosis
VI. Serological Tests (Abs Detection)
For diagnosis of deep mycoses
• Precipitation reaction
• Agglutination reaction
• Inert particle agglutination
• Electrophoretic tests
• Complement fixation
• Indirect fluorescent antibody
• Immunosorbent assays
Fungi : Lab Diagnosis
V. Serological Tests (Ags Detection)
• Cryptococcal antigen in CSF
• Galactomannan in invasive Aspergillosis
• Beta-glucan & D-arabinitol in Candia albicans
Fungi : Lab Diagnosis
IV. Histopathology
• Fungi occur in tissues as one of the following:
1. Yeast Cells
o Intracellular small yeasts like Histoplasma
capsulatum
o May have a large distinguishing capsule as in
Cryptococcus
2. Sporangia
3. Hyphae
o May be brown or colorless
4. Granules
o Are tightly packed hyphae
o Combination of yeast & hyphae as in candida
Fungi : Lab Diagnosis
V. Examination under Wood’s light
• Long-wave UV rays
• When come in contact with mycotic area
of skin or hair produce fluorescent
colours
• Occurs in some mycotic infections only
VI. Fungal Skin Tests
• Has no diagnostic value
• Mainly used for epidemiological studies
• Does not differentiate between active
and past infection
• Observed by induration and swelling
• e.g. Histolasmin test & Trichophytin test
Antifungal drugs
• Fungi closely resemble human cells
• Use the same mechanisms to synthesize
proteins & nucleic acid as higher animals
• Difficulty in selective toxicity
• So relatively few antifungal drugs for
systemic use
• The available systemic drugs are quite
toxic
Antifungal drugs :
Mechanisms of Action
3. Inhibitors of cell 1. Inhibitors of plasma
wall synthesis membrane synthesis
Echinocandins Azoles
Allylamines
2. Inhibitors of plasma
4. Inhibitors of nucleic membrane function
Fungal Cell Polyenes
acid synthesis
Flucytosine
5. Inhibitors of
Cell division
Grisofulvin
Antifungal drugs: Polyenes
• Bind to ergosterols, disrupt the cell membrane, leakage
of the cytoplasm leading to cell death.
1. Nystatin
• Is fungistatic
• Not absorbed from GIT so is used locally only
• Used in treatment of oral & vaginal candidiasis
• Cannot be used in systemic fungal infections
• Has cross-resistance to amphotericin B
2. Amphotericin B
• Is fungicidal
• The most effective for serious fungal infections
• Is used systemically (orally & IV)
• Has toxic effects on nephritic tissues
• Liposomal preparations are less toxic but very
expensive
Antifungal drugs
Azoles
• Interfere with ergosterol synthesis,
leading to defective cell memebrane
A. Imidazoles
• Ketoconazole
• Clotrimazole
• Miconazole
• Mostly used locally to treat yeast, molds &
dermatophytes
• Systemic use is restricted due to
hepatotoxic & antispermatogenesis effects
Antifungal drugs
Azoles
B. Triazoles New Triazoles
• Fluconazole Voriconazole
• Itraconazole Genoconazole
• Have same mode of action like imidazoles
• Are less toxic than imidazoles
• Are used to treat systemic infections
• Fluconazole crosses blood brain barrier and
is used in treatment of cryptococcal
meningitis
Antifungal drugs
Allylamines
• Naftifine
• Terbinafine
• Inhibit an enzyme in the pathway of
ergosterol synthesis
• Are used locally for dermatophytosis
• Terbinafine can be taken orally
Antifungal drugs
Griseofulvin
• Inhibits fungal cell division
• Is fungistatic
• Concentrates in dead keratinized layers of
skin
• Active against dermatophytes only
• Used in skin & nail infections
• Taken orally for months
• Has side effects on stomach
Antifungal drugs
Flucytosine
• Inhibits DNA synthesis
• Effective against yeasts
• Not effective against most molds
• Used in treatment of systemic yeast
infections
Echinocandins
• Caspofungin
• Acts on fungal cell wall
• Used against yeasts & molds
Topical Antifungal Drugs
Polyenes
• Nystatin
Azoles
• Clotrimazole
• Miconazole
• Ketoconazole
Allylamines
• Terbinafine
Antifungal Susceptibility testing
• Practically there was no need for
antifungal susceptibility testing because:
A) Limited number of antifungal drugs
B) Problems associated with antifungal
susceptibility testing