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241 Cards in this Set
- Front
- Back
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what are yeasts, appearance
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unicellular, budding, sexual or asexual reproduction, colonies slimy or mucoid, much like bacteria
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what are molds, appearance
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filamentous, long chains of cells called hyphae, mass of hyphae called mycelium, molds are downy, fluffy, cottony
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what is a mycelium
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mass of hyphae
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what are conidia
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spores
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in dimorphic species, what temp are mycelial forms around
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<or=30 C
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in dimorphic species, what temp are yeast forms or yeast-like forms around
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>35C
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name dimorphic fungi (7)
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Histoplasma capsulatum
Blastomyces dermatidis Coccidiodes immitis Paracoccidioides brasiliensis Sporothrix schenckii Black molds Penicillium marneffei |
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fungal
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tinea versicolor- example of Superficial or Cutaneous Mycoses
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fungal
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chromoblastomycoses: example of subcutaneous Mycoses
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histoplasmosis
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for review
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dimorphic
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histoplasmosis
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dimorphic fungus
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histoplasmosis on gms
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where endemic: histoplasmosis
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ohio river valley
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where found: histoplasmosis
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Bird and bat droppings in soil, caves, ohio river valley
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dimorphic appearance of histoplasmsois and life course in human
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thermally dimorphic: in the environment it grows as a brownish mycelium
inhaled microconidia convert into yeasts in alveolar macrophages go through lymphatics - walled off and calcified in immunocompetent host; in immunocompromised have disseminated dz |
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frequent agar plate for use of evaluating fungi
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Sabouraud's dextrose agar or Sab glucose agar
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what fungus
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aspergillus
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what special containers are needed for fungi
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basically anything
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what is the value of adding chloramphenicol to SAB plate
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inhibits bacterial growth and helps isolate slower growing fungi
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what kind of plate might this be
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SAB
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Selective SAB with choro and cycloheximide alternative names
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Mycosel/Mycobiotic
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use of Mycosel/Mycobiotic plates
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dermatophytes
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use of Selective SAB with choro and cycloheximide
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dermatophytes
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Blood brain heart infusion agar with or without
antibiotics |
useful for looking for systemic fungi
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why add cyclohexamide to an agar
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suppresses saprophytic molds (molds that live on decaying organic matter) to allow for growth of slower growing systemic molds
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****what does cyclohexamide also suppress (besides its intended saprophytic molds) (5)
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Trichosporon beigelii
Candida tropicalis Cryptococcus neoformans Yeast of Blastomyces Yeast of Histoplasma "cyclone TraCCYY" missed the boat |
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what kind of plate is this, what's it looking for
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systemic fungi
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old fashioned way to identify that a fungus truly dimorphic (now done molecularly)
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- usually transferring mold to yeast (but could go the other way)
plate from SAB to blood infusion and v/v and go into cold, rm temp, back and forth for weeks to convert |
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what is the basic flow of how to identify a fungus
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1. Direct preparation
2. Inoculate media/ incubate at 30*C for 4 wk – best to grow mycelial phase of dimorphics 3. Observe for yeast or mold growth 4. Do proper tests for identification a. Mold – observe microscopically using Lactophenol cotton blue b. Yeast – do manual or automated chemical tests |
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what are the three direct preparation steps to ID a fungus
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Gram stain
Calcofluor white stain Impression smears – GMS or PAS |
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why do we Inoculate media and incubate at 30*C for 4 wk –
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best to grow mycelial phase of dimorphics
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when evaluating molds and doing final step of ID after isolation, what do you use
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Mold – observe microscopically using Lactophenol
cotton blue |
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what is a lactophenol cotton blue
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phenol, which will kill any live organisms
lactic acid which preserves fungal structures cotton blue which stains the chitin in the fungal cell walls. |
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what kind of test is this
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Calcofluor white - designed to replace KOH, special product that enables yeast or hyphae to fluoresce
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when is a KOH test performed
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to distinguish between dermatophytes and Candida albicans symptoms (vs. no fungi)
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what are the three categories of dermatophytes
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Trichophyton (found in skin, nail, and hair infections), Epidermophyton (skin and nail infections), and Microsporum (skin and hair infections)
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what kind of test is this and what is it designed to look for
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lactophenol cotton blue
to look for molds (great for myelicum) |
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defunct test for fungi
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Exoantigen immunodiffusion tests
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DNA probe test - where are they most useful
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to confirm dimorphics
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what
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blastomycoses
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association Mucicarmine
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crytococcus neoformans - muco polysaccharide capsule
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fungi
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cryptococcus neoformans
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fungus
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cryptococcus neoformans
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are grams stains better for yeast or mycelial forms
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yeast more than mycelial fungi
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where is histoplasmosis capsulatum found
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eastern half us
mexico caves Bat guano and bird droppings (Starlings) |
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what are the sx of histoplasma capsulatum
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Chronic progressive pulmonary,
cutaneous/mucocutaneous, Chronic systemic (RES system) or acute fulminating systemic disease (fatal) |
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what
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hhistoplasma capusulatum variant duboisii
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fungus - spore form
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histoplasma capusulatum
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what is special about the Histoplasma capsulatum var. duboisi
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found in Africa
more cutaneous and bone manifestations |
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what serologic tests might be mildly useful for histoplasma capsulatum
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Complement fixation test - only good if immune system
Immune diffusion – H and M bands |
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if can't culture, how detect histoplasma capsulatum
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Direct antigen detection in urine (eia for ag in urine); Disseminated and chronic pulmonary disease, can work in immunosuppressed too
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how do you culture histoplasma capsulatum
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at 30*C; SLOW!! 2 – 8 weeks
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appearance of histoplasma capsulatum on SAB plate
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White to brown mycelium , cottony
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what
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histoplasma capsulatum
mold phase tuberculated macroconidia that are large and round (8 – 16 uM) and small microconidia (Microconidia are infectious particle) |
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what is a histoplasma capsulatum look alike fungus
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Sepedonium
not dimorphic macroconidia same but no microconidia |
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what
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histoplasma - macro and microconidia
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what does histoplasma capsulatum look like at 35C in lab
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small yeast, round to oval,
always consistent in size and shape (2 -4 uM) narrow neck in budding |
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what
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2-4uM histoplasma, consistent size and shape, narrow neck budding
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histopathology of histoplasmosis (3)
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- caseating and noncaseating granulomatous inflammation
- calcified lesions - Acute disease in the RES system with intracellular budding yeast forms (2 – 4 mM) |
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what
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intracellular yeast forms (2-4um) of histoplasma
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cf histo and cryptococcus in morphology
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histoplasma - consistent in size and shape
cryptococcus all over the place in size |
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does histoplasma capsulatum have a capsule
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not a true capsule like cryptococcus neoformans
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what
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Leishimania - protozoan parasite; kinetoplast, transmitted by sand fly
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what
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leishmania small, round bodies 2–4 μm in diameter with indistinct cytoplasm, a nucleus, and a small, rod-shaped kinetoplast. (contrast to histoplasma which is 2-4 um but without kinetoplast)
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what
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Histoplasma capsulatum var duboisi
8-10 um |
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what should you think of if you see narrow based budding, 8-10um uniformly sized yeast, tuberculate conidia
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Histoplasma capsulatum var duboisi
8-10 um |
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where is blastomycosis dermatitidis endemic
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eastern USA (esp southeastern)
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clinical fx of blastomycosis
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- pulmonary
- skin -bone |
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growth of blastomycosis
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Culture at 30*C
Takes 2 – 3 weeks to grow |
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on plates
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Fluffy white – buff colored prickly
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what
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broad based budding- Large mother yeast cells averaging 8 – 20 uM or larger.Round to oval
Thick, double contoured double refractile cell walls Single broad based buds |
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what
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blastomycosis -note multiple nuclei
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what
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Pear shaped conidia like lollipops
blastomycosis |
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Pear shaped conidia like lollipops
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blastomycosis
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what
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blastomycosis, Large mother yeast cells averaging 8 – 20 uM or larger.Round to oval
Thick, double contoured double refractile cell walls Single broad based buds |
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blastomycosis Look alike fungus --
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Chrysosporium spp.
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what
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Chrysosporium spp. - not dimorphic, grows in 3-4 days (not weeks) but supposed to be look alike to blastomycosis, grows at 30 and 37
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what
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blastomycosis - pear shaped, lollipop, at 30 C
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fx of blastomycosis on histopathology
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Mixed pyogenic and granulomatousinflammation
Microabscesses Broad based budding yeast cells |
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what
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blastomycosis - rather obscure but notice thick double contoured wall, with "retraction" internal to that. in this the center is somewhat degraded
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where is coccidiodes immitis found
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Found in SW USA, Mexico – areas known as the
Sonoran life zone (cooler part of desert) – desert sands |
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how get coccidiodies
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inhalation of desert sand
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alternative names for coccidiodes
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Valley fever, desert rheumatism, valley bumps
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clinical types of infections: coccidiodes
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Six types of infections:
1) asymptomatic 2-3) pulmonary - focal or diffuse 4) cutaneous 5)meningitis 6)disseminated |
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those at risk for disseminated coccidiodes (3 groups)
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Higher incidence of disseminated in HIV
darker skinned ethnic groups pregnancy |
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growth of coccidiodes
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Culture at 30*C; Takes 2 – 3 days to grow
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appearance on SAB plate: coccidiodes
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White wooly mold
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what on SAB
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blastomycosis dermatidis
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word of caution about coccidiodes
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Very infectious to laboratory personnel
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description of coccidiodes hyphal form
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Septated hyphae with chains of thick walled
barrel shaped arthroconidia with dead cells in between |
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what
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coccidiodes immitis (cause of cocciomycosis)
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Which fungal stain binds to cellulose and chitin present in fungal cell walls, causing fluorescence under ultraviolet light
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calcofluor white
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correct ratio of India ink:CSF that should be placed on a slide
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1:1
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prganism that demonstrates budding yeast cells with wide capsules in an India ink preparation of spinal fluid is probably _____
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cryptococcus neoformans
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best test for cryptococcus neoformans in CSF
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Latex bead antigen agglutination test of the CSF
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formation of germ tubes is s/o
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candida albicans
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what is birdseed agar for
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Cryptococcus neoformans is the only yeast that produces brown colonies (due to melanin production) on birdseed agar
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what
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cryptococcus neoformans - brown colonies on birdseed agar
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phenol oxidase positive
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cryptococcus
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what fungal organism can be associated with catheter-related sepsis
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Malassezia furfur
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fx of Malassezia furfur
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requires lipids from olive oil for growth, and is a small yeast with a wide bud
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what agar is useful at detecting mixed yeast populations
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Chromogenic agar (best at 35-37)
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what candida species doesn't produce germ tubes (good neg ctl for germ tube studies
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Candida tropicalis
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look alike to coccidiodes
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Look alike fungus = Malbranchea species
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what
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White wooly mold
coccidiodes |
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interesting point about dimorphism in coccidiodes
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not dimorphic in lab (same at 30 as 37) but in humans, dimorphic
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histopathology of coccidiodes
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Thick walled spherules (10 – 80 uM) with
endospores All stages of development and fragmented spherules Granulomatous inflammation with caseation |
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two dz that you need to distinguish from coccidiodes
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Rhinosporidium seeberi spherules – larger – 100 – 300 uM in size
Myospherulosis other things make spherules |
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what
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coccidiodes spherules with endospores
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what
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coccidiodes
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what
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coccidiodes
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what
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coccidiodes
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what
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rhinosporidium (bigger) 100 – 300 vs. 10 – 80 uM of coccidiodes- also usually found in oral/nasal (not in lung or deep tissue like coccidiodes)
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what is myospherulosis
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fungi-like spherules composed of erythrocytes damaged by endogenous and exogenous fat
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what
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myospherulosis
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what is another name for paracoccidioses brasiliensis
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south american blastomycosis
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sx of paracoccidiodes
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Pulmonary – primary disease
Can disseminate – muco-cutaneous, lymphatics, CNS, adrenals, GI Hallmark – facial involvement - nasal |
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paracoccidiodes culture
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30C kinda looks like blastomycosis but won't have to worry about that
Culture @ 37*C Slow (3- 4 weeks) waxy coral-like yeast ***Large yeast (10 – 20 uM – up to 60uM) with multiple daughter buds “Mariner’s wheel” |
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what
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mariner's wheel - multiple budding yeast
paracoccidiodes |
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histopathology of paracoccidiodes
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Mixed granulomatous and pyogenic inflammation
Paracoccidiodomas as old coin lesions With polarized light – see maltese cross formation Observe Mariner’s wheel yeast cell – multiply budding large yeast |
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what
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madura foot - known as mycetoma
three sx Swollen lesions usu on extremities Draining sinuses Sulfur granules traumatic implantation (from soil) |
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98% of mycetomas are caused by what
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bacteria! gram + filamentous
Nocardia: brasiliensis and N. asteroides Streptomyces spp Actinomadura Actinomyces israelii (anaerobic bacteria) |
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what
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nocardia
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partial acid fast stain - bug and chemicals of stain
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nocardia
Positive Carbol Fuchsin (like usual AFB) but decolorizes with H2So4 (instead of HCL like most AFB) Potassium permangenate (?) |
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what stains will nocardia be negative for that you might think to try
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its a partial acid fast organism and thus is negative with HCL-using stains like ziehl-nielson, kenyon or rhdamine-rhodamine (?)
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what
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nocardia
3 – 5 days Dry Crumbly Musty smell |
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what is a eumycotic mycetoma
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traumatic transplantation
black molds: Pseudallescheria boydii Maduerella Curvularia Exophiala others |
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what
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black mold: black on reverse too
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what
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curvularia
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what is chromoblastomycosis
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Wart like lesions
Sclerotic bodies in tissue Growth of dematiacious fungi ex.Fonsecaea, Phialophora, Cladosporium, Exophiala, and Rhinocladiella |
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madura foot vs. chromoblastomycosis
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look really similar at end stage - chromoblastomycosis is supposed to be more warty
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three patterns of sporulation of dematiceous fungi
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Rhinocladiella – like sporulation
Phialophora like sporulation Cladosporium like sporulation |
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what
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Rhinocladiella – like sporulation
reminds me of pine branches |
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what
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Phialophora like sporulation
looks like blackberries |
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what
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Cladosporium like sporulation
maybe like forsythia? worry about this one chromoblastomycosis to brain |
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what
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sclerotic body, means chromoblastomycosis, copper penny
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diagnostic feature in chromoblastomycosis
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sclerotic body
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diagnostic feature in mycetoma
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sulfur granules
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what
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sclerotic body
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what
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sclerotic body
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name 3 subcutaneous fungal infections
|
mycetoma
chromoblastomycosis sporotrichiosis |
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what
|
sportotrichiosis
Initial skin lesion w/wo ulceration Lympho-cutaneous spread (LN enlargement)– bone – systemic Pulmonary - CNS (inhaled) |
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explain lab growth patterns of sporothrix
|
Dimorphic dematiacious mold
- 30*C rapid growth (3 -5 days) - turns brown to black over time - septate hyphae with conidia in daisy wheel - At 37*C small oval yeast cells, elongated 2 – 5 uM, cigar bodies |
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what
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sporothrix yeast at 37, black mold at 25
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what
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daisy-like arrangement sporothrix
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what
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sporothrix - daisy like
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what does sporothrix look like on histology
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Pyogenic – to – granulomatous inflammation
hard to find yeast in tissue Asteroid body known as Splendore Hoeppli phenomenon |
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what are you likely to get in tissue for sporothrix
|
splendore-hepaly - no yeast in tissue, but IF found, cigar (ovoid, elongated yeast)
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where do you see rhinosporidosis, sx
|
Water and fish disease in India, Brazil, etc
Chronic granulomatous infection of mucous membranes, mass lesions Endospores can be mucicarmine positive remember really big spherulo |
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nasal
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rhinosporidiosis - note larger spherules than coccidiodes
|
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what is unique about candida
|
Endogenous pathogen
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fungus associated with catheter
related infections in children |
Candida parapsilosis
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association with Candida parapsilosis
|
catheter
related infections in children Parapsilosis, plastic, puer/puella |
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most common candida species
|
c. albicans
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what three candida species may be resistant to fluconazole
|
Candida glabrata, kruseii, and tropicalis
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growth characteristics of Candida in lab
|
Rapid growth = 24 – 48 hours
Bacterial like colony – pasty white Budding yeast – oval @ 7 uM in size and form pseudohyphae (look like sausage links) |
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which candida does not form pseudohyphae
|
Candida (Torulopsis) glabrata is @ 4 uM in
size and does NOT form any pseudohyphae (half the size) - tiny yeast |
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tiny yeast (2)
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histo and C. glabrata
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two tests to id C albicans
|
1) germ tube +
2) Produce chlamydospores on cornmeal agar |
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what other candida is germ tube positive and how distinguish from C. albicans
|
Candida stellatoidea also germ tube positive
but C. stell sucrose positive, while C. albicans is sucrose negative |
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which bug(s) is/are this
|
candida albicans (sucrose -) or c. stellatoidea (sucrose+)
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germ tube test - how long incubate
|
4 hours - if longer, get false positives
incub in plasma |
|
what
|
Positive for Chlamydospores = C. albicans
cornmeal agar, resting spore (protective spore) terminal chlamydospore Incubation at 30*C for @ 48 hours – observe microscopically |
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on histopath for Candida glabrata
|
smaller yeast cells and no pseudohyphae - extracellular
(differentiate from histoplasma) |
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histopath for candida
|
Pyogenic to granulomatous
Usually observe yeast cells, pseudohyphae and/or hyphae appearing structures |
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what is tinea versicolor caused by
|
Malassezia furfur
|
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|
common disease with malezzeia furfur
|
Skin: macules, papules, patches,
plaques on chest back and shoulders with either hypo or hyper pigmentation |
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|
what is a bad form of Malessezia furfur
|
Fungemia in neonates on IV lipid feeding in central lines
|
|
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growth characteristic for Malessezia furfur
|
Lipophilic yeast – required for growth
Small budding yeast 2 – 4 uM with collarette add virgin olive oil to it |
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what
|
M. furfur, spaghetti and meatballs
|
|
|
dermatophytes, tineas, ringworms (hair, skin, nails)
|
Microsporum
Epidermophyton Trichophyton |
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how to dx dermatophytes
|
clinical, KOH prep
|
|
what
|
koh, test
-skin scraping -add Koh (dissolve skin, nail, hair) +/- heat - coverslip and see |
|
what
|
microsporum canis
@ 30*C, one week note: white with yellow edge creeping out |
|
what
|
microsporum canis
Microsporum canis - dog/cat ringworm Tuberculate macroconidia – few micro +hyaline hyphae |
|
what
|
Microsporum gypseum colony @ 30*C at one
week only beige ? |
|
what
|
Microsporium gypseum - soil
|
|
what
|
Trychophyton rubrum colony – red diffusible pigment
on REVERSE |
|
what
|
numerous MICROconidia - lots of shapes, pencil shaped Macroconidia
|
|
|
main cause tinea capitis ring worm in children
|
Trichophyton tonsurans -
|
|
what
|
Epidermophyton floccosum - khaki green color colonies
|
|
what
|
Epidermophyton floccosum
Beaver tail spores – no microconidia |
|
|
where find cryptococcus neoformans
|
Ubiquitous saprophytic yeast assoc. with pigeon, chicken or turkey droppings
inhale it Disease – CNS, blood - can disseminate (meningitis) |
|
|
who is most associated with getting cryptococcus neoformans
|
immunocompromised host - HIV
|
|
|
cryptococcus growth characteristics in lab
|
Irregular sized (2 – 20uM) yeast cells**** round
Polysaccharide capsule*** India ink – negative staining method*** Cryptococcal antigen test – Ag in CSF and blood Daughter cells attached by narrow thread Great variability in size and shape Grows well on all agars – mucoid colonies due to capsule formation Urease + Inositol + |
|
|
what population is crypto not in
|
kids
|
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two chemical tests to know for cryptococcus neoformans
|
Urease + Inositol +
|
|
|
only yeast inositol+
|
crytococcus
|
|
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source of cryptococcus
|
poor pigeon
|
|
phase contrast who
|
round, variably sized
cryptococcus neoformans |
|
what
|
cryptococcus
Mucoid beige |
|
what
|
cryptococcus
|
|
what test, what bug
|
india ink, negative (no) staining of capsule on cryptococcus, can get false haloes on lymphs so ideally get buddng cell
|
|
what
|
cryptococcus on india ink
|
|
what
|
slant where pink is positive for urease, yellow is negative
cryptococcus is urease + |
|
what
|
birdseed agar, chocolate brown colonies = cryptococcus
the background white on this are candida the only yeast to turn brown on birdseed agar is cryptococcus |
|
|
opportunistic fungal pathogens
|
Filamentous molds, septate, usu branching
Hyalohyphomycetes – hyaline (clear) Phaeohyphomycetes – black or dematiaceous Saprophytes in soil, plants, nature |
|
|
most common Hyaline Fungi
|
hundreds
Aspergillus, Penicillium, Fusarium, Paecilomyces, Trichosporon, Geotrichum, Scopulariopsis, Acremonium, |
|
|
aspergillus
|
Hyaline
Septate Branching at 45 degree angle |
|
|
sx/clinical aspergillus
|
Vasoinvasive, thrombosis, infarctions
Supperative inflammation |
|
|
Main look alike for aspergillus
|
Pseudallescheria boydii
|
|
|
most common three aspergillus
|
Aspergillus fumigatus
Aspergillus flavus Aspergillus niger |
|
|
how long for aspergillus spp to grow
|
Grow in 48 – 72 hours
|
|
what
|
aspergillus fumigatus
green carpet |
|
what
|
aspergillus fumagatus
30, green colony, spores are "head up" like a broom |
|
what
|
aspergillus flavus
|
|
what
|
aspergillus flavus
sunburst |
|
what
|
aspergillus flavus
|
|
what
|
aspergillus flavus
|
|
what
|
aspergillus fumagatus - like mop
|
|
what
|
aspergillus niger - salt and pepper
|
|
what
|
aspergillus niger - see black retained on lactophenol cotton blue; head goes all the way around
|
|
what
|
aspergillus
open cavity to have fruiting body fruiting head and dichotamous branching Branching at 45* angle, septate hyphae |
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define dichotamous
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continous
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what
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Fusarium species – Fucshia color
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clinical fx of fusiarium
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Associated with kerititis and disseminated
Infection in BMT |
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what population should you associated with fusarium
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BMT population
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what
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Fusarium
|
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what
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Fusarium
banana boat |
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what fungus is associated
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fusarium - keratitis
moisture - humid |
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what
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penicillium
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what
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bony hand, broom
penicillium (not aspergillus - more like mop) |
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can penicillium cause disease
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yes, skin, nail, disseminated
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what
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Red diffusible pigment of P.marneffei
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only species of Penicillium that is almost always pathogenic
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P.marneffei
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two fungi with red diffusable pigment
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P.marneffei and T. rubrum (but Penicillium is green but T. rubrum is white)
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p. marneffei in clinical settings (2)
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- southeast asia, skin infections
- lung, AIDs |
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what
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p. marneffei
|
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what is only penicillium, dimorphic
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p. marneffei yeast like in tissue, hyphal at 30
|
|
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three black molds that can disseminate
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Phaeohyphomycosis
Eymycotic mycetoma Chromoblastomycosis Phaeohyphomycosis |
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what
|
alternaria
|
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what
|
hand grenade
alternaria |
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what
|
bipolaris- bad actor
sinus, or innocuous lesion in leg then can just march on |
|
what
|
bipolaris
|
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what
|
curvularia
|
|
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alternative name for Pseudallescheria boydii
|
Teleomorph Scedosporium apiospermum
|
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what is Pseudallescheria boydii
|
like aspergillus in tissue (not in lab), vasoinvasive
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|
|
three plate pushers
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Absidia, rhizopus and mucor - grow so fast (within 24 hours) can push the lid off the plate
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what
|
zygomyces
90* angle branching, aseptate, ribbon like, SACS weak - so need to mince not grind |
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need to differentiate between absidia and rhizopus
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based off where sac is... still not clear... for review
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diabetic with rhinocerebral involvement
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mucor
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