Must To Know Mycology
Must To Know Mycology
Must To Know Mycology
Molds Filamentous
Room temperature
Yeasts Round/tissue form
37’C
Chitin Fungal cell wall
Ergosterol Fungal cell membrane
Hyphae Aerial/vegetative/substrate
Spores Reproductive
Sporangiophore Aseptate hyphae
Conidiophore Septate hyphae
Sexual spores Ascospores
Basidiospores
Oospores
Asexual spores Chlamydospores: rounding terminal hyphae or intercalary (within)
Blastospores: budding off (yeasts)
Conidia (macro/micro): dermatophytes
Arthrospores: barrel-shaped, produced from fragmentation of mycelium
10% KOH preparation Skin and hair
Hyphae, yeasts, spores
Dissolves tissue
Calcofluor white Fluorescent dye
Stains the chitin (cell wall)
Green
Giemsa/Wright’s stain H. capsulatum: the only fungi seen in blood smear
Periodic Acid Schiff Glycogen: purple
No glycogen: pink
Gomori silver Black
Dermatophyte test medium SDA + phenol red
Inhibitory mold agar SDA + cycloheximide + gentamicin
Potato dextrose agar Pigment
Cornmeal Tween 80 agar Chlamydospore of C. albicans
Rice medium (+) M. canis
(-) M. audouinii
Staib’s nigerseed agar C. neoformans (black colonies)
Urea agar (+) T. mentagrophytes
(+) C. neoformans
Cottonseed agar B. dermatitidis
Biochemical tests API20C, ID32C
CHROMagar Candida
Germ tube test For unknown yeast
Cornmeal agar Dalmau-Plate
Mannan antigen Candida
Galactomannan Aspergillus
Fungicidal agents Target: Ergosterol
-Amphotericin B: systemic fungi (most common)
-Nystatin
-Azole (Fluconazole): fungistatic
-Griseofulvin (IV): dermatophytes
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AST methods Broth microdilution method
E-test method
Genus-spp ID (molds) Microscopic exam
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(+) Urease
T. tonsurans Thiamine-requiring
Black dot tinea capitis
Microconidia: Balloon-shaped
Colony: Creamy white w/ crater-like depression
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Chromoblastomycosis agents Dematiaceous fungi
Type of sporulation: ID of genus and species
1. Phialophora verrucosa
-Vase-like
-TOS: phialophore
2. Fonsecaea pedrosoi
-Short chain
-w/ 1’, 2’, 3’ conidia
-TOS: Acrotheca
3. Cladosporium carrionii
-Long chain (branching)
-TOS: Cladosporium
Infected tissue: Brown sclerotic bodies (Medlar bodies)
Lesion: Cauliflower-like
Colonies: Dark w/ jet black reverse
R. seeberi Size: 300 μm
Rhinosporidiosis
MOT: swimming
Polypoid masses in nose and pharynx
Tissue form: Sporangium (sac-like structures filled w/ endospores)
L. loboi Lobomycosis
Lesion: Keloid-like subcutaneous nodule involving the extremities
Tissue form: Multiple budding cells in chain (Sampaloc-like/Longganisa-like)
Systemic Mycoses Dimorphic
MOT: inhalation of spores (mold) – infectious
Specimen: Sputum
BSL III = require BSC class II
No person-to-person contact
Colonies: cottony white to brown
Exoantigen test
-B. dermatitidis
-P. brasiliensis
-H. capsulatum
-C. immitis
B. dermatitidis Agent of:
-North American blastomycosis
-Chicago disease
-Gilchrist disease
Pneumonia and skin infections
Mold form: Lollipop in appearance
Yeast form: Single-budding yeast w/ broad based (double centered) – diagnostic
Mickey mouse cap
Culture: Medium w/ cycloheximide
Tissue: KOH
Tx: Amphotericin B
P. brasiliensis Agent of:
-South American Blastomycosis
-Lutz Splendore-Almeida disease
Infects spleen, liver, lymph node, skin, lungs
Mold form: Lollipop in appearance
Yeast form: Multiple budding yeast resembling Mariner’s wheel
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H. capsulatum RES parasite
Agent of Darling’s disease
“Arise of Tutankhamen’s tomb”
USA = endemic dimorphic ffungus
MOT: inhalation of spores from guanos (droppings of birds, starlings/chickens, and
bats)
Mold form: Tuberculate macroconidia
Yeast form: Yeast cells intracellular in macrophages
Culture: Medium w/ cycloheximide (BM, sputum, tissue)
H. duboisii Double cell
Figure of 8
Causes African histoplasmosis
Grows in N2
Sepedonium Tuberculate macroconidia (resembles H. capsulatum)
To differentiate: Sepedonium is monomorphic
C. immitis Agent of:
-Desert fever
-San Joaquin Valley fever
Alkaline soil
High humidity
Lab. acquired infection
Mold form: Barrel-shaped arthroconidia (200μm)
Yeast form: spherule w/ endospores
Culture: medium w/ cycloheximide (30’C)
Not processed ---(send)---> Reference lab
Lab. Diagnosis 1. Direct examination
(Systemic mycoses) a. Histoplasma: Wright’s/Giemsa
b. Blastomyces, Paracoccidioides, Coccidioides
- KOH: tissue form
- PAS
- H&E
2. Cultures
a. SDA = RT’ (mold)
b. BHIA + blood = 37’C (yeast)
3. Immunological tests
a. Coccidioidin
b. Histoplasmin
Exoantigen Test Principle: Double Diffusion
A = B. dermatitidis
HS, HL, F = C. immitis
H & M = H. capsulatum
1, 2, 3 = P. brasiliensis
Opportunistic Mycoses Normal flora (Ex. Candida)
At risk: immunocompromised person
-Candida
-Cryptococcus
-Aspergillus
C. albicans Normal flora: skin, mucous membrane, GIT
Produce yeast and hyphae in vivo
-Germ tube
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-Chlamydospore
-Blastoconidia
-Pseudohyphae
-True hyphae
(+) Sucrose
EMB: feathering colonies
Diseases:
-Thrush
-Diaper rash
-Candidiasis (Old: Moniliasis): cutaneous, oropharyngeal (white patches),
esophagitis, cheilitis, onchomycosis, vulvovaginitis, invasive infection (CNS, blood)
Predisposing factors:
-Prolong antibiotic use especially broad spectrum = disturb pH
-Pregnancy
-DM
-Malnutrition
Lab. Diagnosis:
1. (G) SDA
2. Screening test: Germ tube test
= Organisms + Serum ---(35’C for 2-3 hours)---> (+) Germ tube
3. Confirmatory test: Chlamydospore Cornmeal agar
= C. albicans Inoculate: CMA ---(RT’ for 48-72 hours)---> (+) Chlamydospore
Other Candida spp C. tropicalis = infection on immunocompromised host
C. glabrata = 2nd common spp
C. krusei = hematological disorders
C. parapsilosis = endocarditis
Lab. Diagnosis (Candida) Vaginal pH: 4.5
Rule out vaginosis/Trichomoniasis = alkaline pH
Vaginal discharge = 10% KOH (Whiff’s test)
Fungal culture
(+) Germ tube test C. albicans = (+) Chlamydospores, (+) Sucrose, (G) 42’C
C. stellatoidea = (-) Chlamydospores, (-) Sucrose, (NG) 42’C
G. candidum = (+) Arthrospores
C. dubliniensis = (NG) 42’C, double conidia
Cryptococcus (Filobasidiella) Found in pigeon droppings and soil
neoformans MOT: inhalation of airborne organism
Encapsulated yeast (India ink)
Diseases:
-Meningitis
-Torulosis (Old: Cryptococcosis)
-Pneumonia
Lab. Diagnosis:
-Old: India ink preparation = CSF, pulmonary tissue
-New: Latex agglutination (Ag): capsule
-Biochemical tests:
= (+) Inositol
= (+) Urease
= (-) NO3 assimilation = yellow
= (+) Phenoloxidase = makes the colonies black
= (+) Phospholipase
-(G) Birdseed agar
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= Assimilate creatinine
= Colonies: Yeastlike, mucoid, cream to brown
-Culture: SDA w/o cycloheximide
Tx: Amphotericin B, Fluconazole
Rhodotorula Resemble C. neoformans
Encapsulated
To differentiate: Rhodotorula = pink colonies, (+) Urease, (+) NO3 assimilation
Aspergillus Bread mold
Septate hyphae
Found in nature: soil, plant, air
Vesicles: swollen conidiophores
A. flavus Aflatoxin (toxicoses) = HCC
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MUST TO KNOW IN VIROLOGY
Ether-resistant Naked virus
Ether-sensitive Enveloped virus
Acid-labile Rhinovirus
Acid-stable Enterovirus
Infantile diarrhea Rotavirus
Myxoviruses H&N
Hemagglutination/ Influenza
Hemadsorption Parainfluenza
Mumps
Reye’s syndrome Chicken pox
Influenza
Bronchiolitis in children RSV
Hepatitis Yellow fever
HBV
CMV
Reservoir of Togavirus Birds
dsRNA Reovirus
ssDNA Parvovirus
Tzanck smear HSV
VZV
Latent infection HSV
Rubeola virus Measles
Rubella virus German measles
SSPE Measles virus
PML JC virus (polyoma)
Prion Spongiform encephalopathy
Viroid RNA (HDV)
Human Embryonic Fibroblast CMV
cell
PMKC, MDCK, Hen’s egg Influenza
5th disease Parvovirus B19
6th disease HHV-6
Kaposi’s sarcoma HHV-8
Filoviridae Marburg and Ebola
Coronavirus SARS
Influenza virus Bird’s flu = H5:N1
Use cotton swabs Virus
Delay of 3 days 4’C
Delay of 4 days -70’C, not -20’C
CPE Tissue culture
33’C Rhinovirus
EM Negative stain
Gold, silver stain
PTA
Roller drum Hold cell culture tube
Dmitri Iwanowsky Father of virology
Nucleic acid genome DNA or RNA
Capsid Protective protein coat
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Envelope Lipid derived from host cell membrane
Lab. Diagnosis:
1. Tzanck smear: multinucleated giant cell w/ cowdry type A inclusion (HSV, VZV)
2. IFT: method of choice
EBV Infects B cells (CD 21)
(+) Heterophile antibodies
Agent: IM
Burkitt’s lymphoma and nasopharyngeal carcinoma
Lab. Diagnosis:
-Not cultured
1. Heterophil Ab test: monospot
2. EBV specific Ab test: EBVCA IgM, EBNA
3. Hematology: Downey cells
CMV MOT: Transplacental, oral, sexual, blood, tissue transplant
#1 congenital infection
Mononucleosis-like but (-) Heterophile Ab
“Owl’s eye” inclusion (PAP or Giemsa) = urine/tissue
Culture:
-HDF cell = method of choice
HHV-6 Roseola infantum
6th childhood disease: skin rash
Reactivation in AIDS:
-Encephalitis
-Pneumonitis
-Chorioretinitis
HHV-7 No known disease
HHV-8 Kaposi’s sarcoma (HIV)
Polyomavirus BK virus = renal disease
JC virus = PML: Progressive Multifocal Leukoencephalopathy
Papillomavirus Tropism for squamous epithelial cells = Warts (fingers, sole, face)
Cervical, squamous cell, vulvar, penile cancer
Condylomata acuminate = anogenital warts
Adenovirus #1 Viral conjunctivitis
Serotypes:
1.) 3, 4, 7, 21 = acute respiratory disease
2.) 8, 19 = epidemic keratoconjunctivitis
3.) 11, 21 = hemorrhagic cystitis
4.) 40, 41 = infantile gastroenteritis
Hepadnavirus Acute and chronic hepatitis B
#1 Blood borne disease
HCC
Co-infection/superinfection w/ HDV
Lab. Diagnosis:
-ELISA
-PCR
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-Markers:
a. HBsAg
b. HBeAg
c. anti-HBc
d. anti-HBs
e. anti-HBe
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Lab. Diagnosis: EM
Retrovirus Reverse transcriptase: RNA DNA
1. Lentivirus
-HIV: AIDS
2. Oncornavirus
-HTLV-I: Human T cell Leukemia
-HTLV-II: Hairy Cell Leukemia
Lab. Diagnosis:
a. Serology:
-ELISA (screen)
-WB (confirm)
-PCR: HIV-DNA
Arenaviridae Arena-Sandy hemorrhagic fever
1. Lymphocytic choriomeningitis (LCM) virus
2. Junin virus = Argentina
3. Machupo virus = Bolivia
4. Lassa fever virus = Zoonotic (rats)
HAV Picornavirus (Enterovirus 72)
Infectious hepatitis
HBV Hepadnavirus
Serum hepatitis
HCV Flavivirus
Non-A, Non-B/post-transfusion hepatitis
HDV Viroid-like
Require HBV infection
HEV Calicivirus
Water-borne
HGV Flavivirus
Blood-borne hepatitis
Prion-caused diseases (Slow 1. Scrapie = sheep/goat
disease) 2. Kuru
3. Creutzfeldt-Jakob disease (CJD) = man
4. Gerstmann-Straussler Scheinker syndrome (GSS)
5. Bovine spongiform encephalopathy = mad cow disease
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