3. MUST to KNOW in Mycology and Virology
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MEDTECH...
Description
MUST TO KNOW IN MYCOLOGY
M. canis
Fluoresces on Wood’s lamp Agent of dandruff
Mycosel
Medium for dermatophytes Chloramphenicol: Chloramphenicol: inhibit bacteria Cycloheximide: inhibit fungal contaminants
Czapek’s medium
Aspergillus Aspergillus
Germ tube
Forms at 35’C (yeasts)
Candida
Gram (+), oval cells w/ or w/o buds and pseudomycelia
SDA
General culture media
Basic structure of fungi
Hyphae Spore
Cornmeal agar
Chlamydospore
Media w/ olive oil
M. furfur
Hair baiting test
T. mentagrophytes
Trichophyton
Infects hair, skin, and nails
T. rubrum
Red color Teardrop conidia
T. schoenleinii
Favic chandelier (antler)
Coenocytic (aseptate)
Phycomycetes/Zygomycetes
Ascospores
S. cerevisiae
C. neoformans
(+) Nigerseed/Birdseed Nigerseed/Birdseed (+) Urease (+) India ink (+) Phenoloxidase Phenoloxidase
Arthrospores
“TACGeo” Trichosporon beigelii Aureobasidium Aureobasidium Coccidioides Geotrichum candidum
Blastospores
Yeast
10% KOH
Direct microscopic Clearing
LPCB
Lactic acid: preservative Phenol: killing agent Cotton blue: stains the chitin (cell wall) Commonly used stain (fungal culture)
Swab
Unsuitable for fungal culture Suitable for bacteria and viruses only
H. capsulatum
Wright’s/Giemsa stain
Aspergillus Aspergillus
Dichotomous
lec.mt 04 |Page | 96
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C. albicans
Thrush Diaper rash Moniliasis In tissue: mold/yeast form
Dermatophytes
Macroconidia and microconidia
Sputum
Specimen for systemic mycoses
CHO assimilation
ID of yeast Free of CHO
BHIA
Dimorphic fungi
C. immitis
High risk to lab personnel
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 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: capsulatum: the only fungi seen in blood smear
Periodic Acid Schiff
Glycogen: purple No glycogen: pink
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Rice medium
(+) M. canis (-) M. (-) M. audouinii
Staib’s nigerseed agar
C. neoformans (black 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 Aspergillus
Fungicidal agents
Target: Ergosterol -Amphotericin B: systemic fungi (most common) -Nystatin -Azole (Fluconazole): fungistatic -Griseofulvin (IV): dermatophytes dermatophytes
AST methods
Broth microdilution method E-test method
Genus-spp ID (molds)
Microscopic exam
Superficial Mycoses
Non invasive No immune response response from the host MOT: MOT: person-to-person (garment) - M. M. furfur -P. hortai -T. beigelii -P. -P. werneckii we rneckii
M. furfur
Ptyriasis versicolor (An-An) Hypo/Hyperpigmentation Hypo/Hyperpigmentation of skin KOH: budding yeast cells and hyphae PAS: “Spaghetti and meatballs” Lipophilic: require lipids for growth (SDA + olive oil)
Piedra agents
Hair
P. P. hortai
Black piedra Dematiaceous Ascospores (sexual)
T. beigelii
White piedra Cream colored Arthrospores (asexual)
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Dermatophytes
Keratinophilic Tinea or ringworm Endothrix: -T. tonsurans: dandruff (Tinea capitis) -T. violaceum Ectothrix: - M. M. gypseum -M. canis -T. verrucosum
Tinea barbae
Beard
Tinea capitis
Scalp
Tinea corporis
Body
Tinea cruris
Groin
Tinea pedis
Foot (Athlete’s foot)
Tinea unguium
Nails
T. rubrum
Red pigment Microconidia: tear-drop shaped (side) Macroconidia: pencil-shaped Colony: Fluffy white w/ red color reverse
T. mentagrophytes
Microconidia: grape-like (cluster) (+) Hair penetration test: V-shaped (+) Urease
T. tonsurans
Thiamine-requiring Black dot tinea capitis Microconidia: Balloon-shaped Colony: Creamy white w/ crater-like depression
T. schoenleinii
Require thiamine and inositol Cause Favus: chronic hair infection Favic chandelier hyphae
T. verrucosum
Microconidia: clavate/pyriform Macroconidia: rat-tail/string-bean-shaped rat-tail/string-bean-shaped
M. canis
Macroconidia: spindle-shaped, echinulate (+) Wood’s lamp
M. gypseum
Geophilic Macroconidia: oblong (ellipsoidal) w/ blunt/rounded blunt/rounded ends (-) Wood’s lamp
M. audouinii
Anthropophilic Tinea capitis (old) (NG) Rice medium
E. floccosum
Macroconidia: club shaped in pairs
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Subcutaneous My Mycoses
MOT: sk skin tr trauma/prick Habitat: soil Biopsy, granules (PAS, H&E) -Sporothrix schenckii -Madura foot agents -Chromoblastomycosis -Chromoblastomycosis agents - Rhinosporidium Rhinosporidium seeberi -Loboa loboi
S. schenckii
Mold form: flowerette/daisy-like flowerette/daisy-like conidia Yeast form: Cigar-shaped (asteroid) bodies Rose Gardener’s disease -Cord-like multiple subcutaneous nodules Colony: White to black
Madura fo foot ag agents
Maduromycosis: my mycetoma P. P. boydii = boydii = most common cause Madurella Madurella Leptosphaeria Leptosphaeria Actinomycetes Tissue form: Granules Lesion: granulomatous granulomatous lesions on foot w/ multiple draining sinus tracts
P. P. boydii
Cleistothecia Teleomorph/sexual form Scedosporium apiospermum: apiospermum: anamorph/asexual form
Chromobla Chromoblastom stomycosi ycosiss agents agents
Dematiac Dematiaceous eous fungi Type of sporulation: ID of genus and species 1. Phialophora 1. Phialophora verrucosa verrucosa -Vase-like -TOS: phialophore 2. Fonsecaea 2. Fonsecaea pedrosoi 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 Cauliflower-like Colonies: Dark w/ jet black reverse r everse
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) (Sampaloc-like/Longganisa-like)
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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. B. dermatitidis -P. brasiliensis -H. capsulatum -C. immitis
B. dermatitidis
Agent of: -North American blastomycosis 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. P. brasiliensis
Agent of: -South American Blastomycosis 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
H. capsulatum
RES parasite Agent of Darling’s disease “Arise of Tutankhamen’s tomb” USA = endemic dimorphic ffungus MOT: MOT: inhalation of spores from guanos (droppings of birds, starlings/chickens, 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 histoplasmosis Grows in ! N2
Sepedonium
Tuberculate Tuberculate macroconidia (resembles H. (resembles H. capsulatum) capsulatum) To differentiate: Sepedonium is monomorphic monomorphic
C. immitis
Agent of: -Desert fever -San Joaquin Valley fever Alkaline soil
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Lab. Diagnosis (Systemic mycoses)
1. Direct examination examination a. Histoplasma a. Histoplasma:: Wright’s/Giemsa b. Blastomyces, b. Blastomyces, Paracoccidioides, Paracoccidioides, Coccidioides Coccidioides - KOH: tissue form - PAS - H&E 2. Cultures a. SDA = RT’ (mold) b. BHIA + blood = 37’C (yeast) (yeast) 3. Immunological tests a. Coccidioidin b. Histoplasmin
Exoantigen Test
Principle: Double Diffusion A = B. = B. dermatitidis HS, HL, F = C. immitis H & M = H. = H. capsulatum 1, 2, 3 = P = P.. brasiliensis
Opportunistic Mycoses
Normal flora (Ex. Candida) Candida) At risk: immunocompromised immunocompromised person -Candida -Cryptococcus -Aspergillus
C. albicans
Normal flora: skin, skin, mucous membrane, membrane, GIT Produce yeast and hyphae in vivo -Germ tube -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:
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Lab. Diagnosis (Candida (Candida))
Vaginal pH: 4.5 Rule out vaginosis/Trichomoniasis vaginosis/Trichomoniasis = alkaline pH Vaginal discharge = 10% KOH (Whiff’s test) Fungal culture
(+) Germ tube test
C. albicans = albicans = (+) Chlamydospores, Chlamydospores, (+) Sucrose, (G) 42’C C. stellatoidea = stellatoidea = (-) Chlamydospores, Chlamydospores, (-) Sucrose, (NG) 42’C G. candidum = candidum = (+) Arthrospores C. dubliniensis = dubliniensis = (NG) 42’C, double conidia
Cryptococcus (Filobasidiella) (Filobasidiella) neoformans
Found in pigeon droppings and soil MOT: inhalation of airborne organism Encapsulated yeast (India ink) Diseases: -Meningitis -Torulosis -Torulosis (Old: Cryptococcosis) Cryptococcosis) -Pneumonia Lab. Diagnosis: -Old: India ink preparation = CSF, pulmonary tissue -New: Latex agglutination (Ag): capsule -Biochemical tests: = (+) Inositol = (+) Urease = (-) NO3 assimilation = yellow = (+) Phenoloxidase Phenoloxidase = makes the colonies black = (+) Phospholipase Phospholipase -(G) Birdseed agar = 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 Aspergillus
Bread mold Septate hyphae Found in nature: soil, plant, air Vesicles: swollen conidiophores
A. flavus
Aflatoxin (toxicoses) = HCC
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Zygomycosis/ Mucormycosis
Agents: Zygomycetes -Rhizopus -Absidia -Mucor MOT: inhalation of airborne conidia Infections: -Rhinocerebral: Rhino-facial-cranial Rhino-facial-cranial -Lungs, GIT, skin Lab. Diagnosis: -Culture w/ SDA
Penicillium
Brush-like conidiophores conidiophores Colony: white to bluish green, yellow/brown
Fusarium
Sickle or canoe shaped, multiseptate macroconidia macroconidia Colony: Cottony white to pink or purple colony
Phaeo haeohy hyp phom homycos ycosis is
Demat ematia iace ceou ouss fun fungi (Exc (Excep eptt Chro Chrom moblast lasto omycos ycosiis agen agents ts)) - Alternaria Alternaria -Bipolaris -Curvularia -Dreschlera -Exophiala
P. P. jiroveci (Old: P (Old: P.. carinii) carinii)
Old: Old: Prot Protoz ozoa oan n cys cystt (pa (para rasi site te)) bec becau ause se it has has no no erg ergos oste tero roll New: Fungi because because of DNA homology Diseases: -#1 cause of pneumonia in AIDS patient -#1 opportunistic infection in AIDS patient Lab. Diagnosis: -Masson-Fontana silver stain -Mucicarmine: stain for spherule
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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/ Hemadsorption
Influenza 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
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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 PTA
Roller drum
Hold cell culture tube
Dmitri Iwanowsky
Father of virology
Nucleic acid genome genome
DNA or RNA
Capsid
Protective protein coat
Envelope
Lipid derived from host cell membrane
Icosahedral
DNA virus (+) Sense RNA virus
Helical
(-) Sense RNA virus
Complex
Poxvirus
EM (inv (inver erte ted d micr micros osco cope pe))
Viral iral morp morpho holo logy gy
LM
Inclusion bodies: -Negri bodies: Rabies -Councilman bodies: Yellow fever
Direct IF
Viral antigen
Eggs
(G) Pock formation on CAM (Chrorioallantoic membrane)
Primary cell
Passed only once Ex. 1’ monkey kidney cells (fluvirus)
Semi-continuous
Passed 50x Ex. HDF/HEK cells
i
li
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Syncytium form
RSV Measles Rubella (giant cells)
Hemadsorption
Influenza Parainfluenza Measles Mumps
Refractile, round cell
Rhinovirus (33’C)
Multiplication cycle
1. Adsorption = attachment 2. Penetration = entry 3. Uncoating = (-) capsid 4. Eclipse (Synthetic) = replication " transcription " translation " viral CHON 5. Assembly = creation of new virus particle 6. Release
DNA Viruses
“HHAPPPy” Herpesvirus Hepadnavirus Adenovirus Poxvirus Papovavirus (Papilloma, Polyoma, Simian Vacuolating Virus) Parvovirus
Rule: DNA viruses
All are dsDNA except: Parvovirus (ssDNA) All are icosahedral except: Poxvirus (complex) All multiply in the nucleus except: Poxvirus (cytoplasm) All are enveloped except: PAP (Naked)
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Herpesviridae
1’ and latency infection Alpha = HSV-1, HSV-2, VZV Beta = CMV, HHV-6, HHV-7 Gamma = EBV, HHV-8
HSV-1
1’ infection: Gingivostomatitis Latent infection -Herpes labialis -Fever blister -Cold sore -Keratitis Common cause of viral encephalitis Latency site: Trigeminal ganglion
HSV-2
Genital herpes Neonatal herpes herpes Aseptic meningitis Cervical cancer (HPV) Latency site: Sacral ganglion
Lab. Diagno Diagnosis sis (HSV (HSV-1 & 2)
1. Tzanck Tzanck smear: smear: multinu multinuclea cleated ted giant giant cells cells w/ cowdry cowdry inclusio inclusion n 2. Cell culture: most diagnostic -CPE: 1-5 days -Virus ID: IFT 3. PCR
VZV
1’ infection: Varicella/Chicken pox Recurrent/Reactivation: Recurrent/Reactivation: Zoster/Shingles Zoster/Shingles Latency site: Dorsal root of ganglia Association in Reye’s syndrome
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HHV-8
Kaposi’s sarcoma (HIV)
Polyomavirus
BK virus = renal disease JC virus = PML: Progressive Multifocal Leukoencephalopathy Leukoencephalopathy
Papillomavirus
Tropism for squamous epithelial cells = Warts (fingers, sole, face) Cervical, squamous cell, vulvar, penile cancer Condylomata 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 gastroenteritis
Hepadnavirus
Acute and chronic hepatitis B #1 Blood borne disease HCC Co-infection/superinfection Co-infection/superinfection w/ HDV Lab. Diagnosis: -ELISA -PCR -Markers: a. HBsAg b. HBeAg c. anti-HBc d. anti-HBs e. anti-HBe
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Picornaviridase
Acid Resistance: -Enterovirus: acid-resistant -Rhinovirus: acid-stable Site to multiply: -Enterovirus: GIT -Rhinovirus: Nasal Aseptic meningitis
Poliovirus
Poliomyelitis MOT: MOT: fecal-oral, respiratory droplets Anterior horn cells of the spinal cord Vaccines: 1. Salk vaccine = killed/inactivated killed/inactivated virus (injected) 2. Sabin vaccine = live attenuated virus (oral)
Coxsackie A
Herpangina
Coxsackie B
Pleurodynia
ECHOvirus
Enteric Cytopathic Human Orphan virus Aseptic meningitis
HAV
Enterovirus 72
Rhinovirus
Acid-labile (33’C) Common cause of common colds
Orthomyxoviridae
Hemagglutinin (H) and Neuraminidase (N) Antigenic changes: a. Shift: genetic reassortment (pandemic) b. Drift: point mutation mutation (epidemic) (epidemic)
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Parainfluenza virus
Croup: Laryngotracheobronchitis Specimen: NPA/W NPA/W Culture: PMK, LLC-MK2 ID: Hemadsorption, IF, EIA Tx: Aerosolized ribavirin No vaccine
Mumps virus
Parotitis, orchitis, aseptic meningitis (+) in saliva and urine = IFA, EIA, HAI Culture: PMK, HEK, Embryonated egg
Measles (Rubeola) virus/ Morbillivirus
Koplik spots 2nd infection: SSPE (Subacute sclerosing panencephalitis) panencephalitis) Specimen: NPS, urine Culture: PMK (multinucleated cells) Serology: (+) IgM Ab’s
RSV
#1 Bronchiolitis in children Specimen: NPS " DFA, EIA Culture: PMK (syncytia), HDFC Tx: Ribavirin
Henipavirus
Nipah virus: Encephalitis Encephalitis = Pig " Man Hendra virus: Respiratory disease in horses
Togaviridae
Arbovirus Alphavirus Rubivirus
Arbovirus
Arthropod borne (mosquito) Includes:
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Rhabdoviridae
1. Rabies virus (Lyssavirus) -Bullet-shaped -Cause encephalitis encephalitis Lab. Diagnosis: Negri bodies (dog brain) = Seller’s stain 2. Vesicular stomatitis -Cattle, horse, pig
Filoviridae
Filamentous 1. Marburg-Ebola virus 2. Hemorrhagic fever
Reoviridae
1. Rotavirus = Wagon wheel like (EM) 2. Orbivirus = Colorado tick fever
Coronavirus
Club-shaped SARS agent; common colds Gastroenteritis Lab. Diagnosis: EM, IF, EIA
Calicivirus
Norwalk-like virus virus
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Prion-caused diseases (Slow disease)
1. Scrapie = sheep/goat 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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