IDENTIFICATION

OF FUNGUS
Dr. Azimah Abdul Wahab
azimahaw@unikl.edu.my
INTRODUCTION
• Based on transmission fungal diseases are basically
classified into THREE types:
I. AIR BORNE DISEASES (E.g: blastomycosis,
cryptococcosis)
II. DIRECT CONTACT DISEASES (E.g: superficial or
cutaneous mycosis - tinea)
III. OPPORTUNISTIC DISEASES (E.g: Aspergillosis,
candidiasis)
 Mycoses
• Mycoses is fungal infections.
• A fungal or fungus-like disease occurring in animal/human
with compromised immune system
• Inhalation of fungal spores is transmission of the disease.
• Examples:
1) Candidiasis
2) Aspergillosis
3) Cryptococcosis
4) Histoplasmosis
5) Blastomycosis
6) Dermatophytosis

• Multiple types of Mycoses :

❑ Superficial Mycoses
❑ Cutaneous Mycoses
❑ Systemic Mycoses
Candidiasis

➢ Caused by Candida sp. (E.g: Candida

albicans)
➢ Has Ovoid shape or spherical
budding cells.
➢ Cultures are done on Sabouraud’s
Dextrose agar.
➢ Dimorphic (growing as yeast cells
and filamentous hyphae cells) -
Grow predominantly in yeast
phase.
➢ Normal flora - in mouth,
gastrointestinal tract, vagina
Microscopic
appearance
of Candida
sp.
Prominent infection with candida spp.
Aspergillus sp.
➢ Caused by Aspergillus sp.
➢ Eg: Aspergillus fumigatus,
Aspergillus niger
➢ Is a group of moulds
➢ Common in the home,
including bedding
➢ Only a few of these moulds
can cause illness in humans
and animals
Pathogenesis

➢ This mold produces abundant small

conidia that are easily aerosolized
➢ Following inhalation of these conidia,
atopic individuals often develop severe
allergic reactions to the conidial agents
➢ In immunocompromised patients, the
conidia may germinate to produce
hyphae that invade the lungs and other
tissues
CRYPTOCOCOSIS
❑Caused by Cryptococcus
sp.
❑It grow in culture as yeast
❑Find in soil
❑Surrounds by variables
thickness of polysaccharide
capsule
Pathogenesis
 Histoplasmosis
• The fungus lives in the environment,
particularly in soil that contains large
amounts of bird or bat droppings.
• People can get histoplasmosis after
breathing in the microscopic fungal
spores from the air
• most people who breathe in the spores
don’t get sick, those who do may have a
fever, cough, and fatigue
Blastomycosis
• Lives in the environment,
particularly in moist soil and in
decomposing matter
• Transmission – breathing in the
fungal spores
• Some people will develop
symptoms like fever and cough
Dermatophytoses
• Extend deeper into
epidermis, also nails and
hairs.
• Caused by
keratinophilics fungi,
which is known as
Dermatophytes.
• Dermatophytes - Digest
keratin by their keratinases
• Examples: Ringworm and
tinea
SPECIMENS
• Skin scrapping
• Nail
• Hair
• Sputum
• Blood
• CSF
• Tissue Biopsy
 Microscopy
• KOH preparation – It is used as a
primary screening tool.
➢ To dissolve tissue material,
leaving the alkali-resistant fungi
intact.
➢ It helps to visualize fungal
elements but may not
necessarily identify species of
the fungi.
• Recommended concentrations to be
used:
➢ 10% (skin, hair)
➢ 40% (nail)
Result of KOH
Microscopy
• Gram’s stain –
➢ to identify
yeast, or yeast-
like fungus
Microscopy
• Negative stain –
➢ Use india ink or nigrosin
➢ to identify Cryptococcus
sp.
 Microscopy
• Lactophenol Cotton Blue (LPCB) – identification
of the fungal cell walls.
• The solution is clear and blue in color and it is
made up of a combination of three main reagents:
• Phenol: It acts as a disinfectant by killing
any living organisms
• Lactic acid: To preserve the fungal
structures
• Cotton blue: To stain or give color to the
chitin on the fungal cell wall and other fungal
structures
• The stain will give the fungi a blue-colored
appearance of the fungal spores and structures,
such as hyphae.
Histoplasma sp. Dermatophytes Aspergillus sp. Penicillium sp.
 Culture

Bird seed agar Dermatophyte Test

SDA agar CHROM agar
(isolation of Medium (DTM)
(isolation of (identification of
Cryptococcus (to differentiate
fungus) Candida sp.)
neoforman) dermatophytes from
other fungi)
Culture
Condition
Culture
Identification
 Culture
Identification
• Microscopic appearance
➢ Morphology
➢ Staining (E.g: LPCB)
➢ Type of hyphae
(septate/ aseptate)
➢ Type of sporulation
(conidia/ sporangia)
Culture
Identification
 Immunological
Methods

• Antibody detection – ELISA,

Agglutination Test
• Antigen detection – Latex
agglutination test
• Immunohistochemistry
• Metabolite test – Gas liquid
chromatography
• PCR