MYCO L2 W/O TABLES Flashcards by Cardenas Xaira Andrea

respiratory tract secretions (enumerate)

sputum, induced sputum,
bronchial washings, bronchoalveolar lavage, and tracheal
aspirations

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mL required to inoculate each medium of respite tract secretions

0.5mL

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why are Antibacterial antibiotics should be included in the battery of media used in respiratory secretions

to ensure optimal recovery of fungi and prevent
overgrowth by contaminants

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(respiratory secretions) should be included in at least one of the
culture media to prevent overgrowth by rapidly growingmolds

Cycloheximide

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CSF Collected for culture should be filtered through a

0.45-μm membrane filter attached to a sterile syringe or
centrifuged

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(CSF) Cultures should be examined

daily

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(CSF) media used for the recovery should contain

no antibacterial or antifungal agents

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CSF specimens should be processed promptly but if not possible should be

kept af RT or in incubator at 30C

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Specimen recommended for disseminated fungal infections

Blood

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Fungal blood cultures commonly used

biphasic BHIA-
broth and lysis-centrifugation system

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(blood) suggested incubation time

21 days

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(blood) most fungi are detected within the first 4 days of incubation except for

H.capsulatum

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(BLOOD)H.capsulatum require approximately (days of incubation)

10-14 days

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optimal temperature for fungal blood cultures

30C

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usually submitted specimen for dermatophyte culture

hair,skin,and nail scrapings

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(hair ,skin,and nail scrapings) may be obtained by scraping thr skin or nails with a scalpel or blade or microscope slide

Lesions

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(hair,skin and nails) infected hairs are removed by

plucking them with forceps

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(hair ,skin, and nails) how to sotre specimens

should not be refrigerated

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(hair,skin,nails)specimens are plated in a

Mycobiotic or Mycosel agar

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(hair,skin,nails)Cultures shoukd be incubated for how many days before being reported as negative

minimum of 21 days at 30C

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vaginal samples should be transported to the laboratory within - of collection using culture transport swabs

24 hours

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(vaginal) swabs should be kept- in sterile tune for a wet preparation

moist

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vaginal culutres should be screened for yeasts and incubated at

30c for 7 days

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(urine) should be added to media to ensure recovery of fungi

antibacterial agents

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(tissue,bone marrow,and sterile body fluids) cultures should be incubated at

30C for 21 days

Recommended battery of media for optimal recovery of fungi

Media with and without cycloheximide Media with and without an antibacterial agent

culture dishes should comtain - and placing them in a humidified incubator

at least 40 mL of agar

cultures should be incubated at RT, or

30°C, for 21 to 30 days and examined at least three times weekly

addition if this to media traditionally has been advocated to inhibit the growth of contaminating molds and bacteria, respectively

0.5 μg/mL of cycloheximide and 16 μg/mL of chloramphenicol

antibacterial agents that gives better result

5 μg/mL of gentamicin and 16 μg/mL of chloramphenicol

temporary mounts

10-20% KOH Lactophenol cotton blue India ink or Nigrosin Calcofluor white stain

Traditionally recommended method

10-20% KOH

KOH preparation is left at to allow clearing of epidermal cells

RT for 5 to 10 minutes

addition of these in KOH enhance visibility of fungal elements

lacto phenol (anioine) blue or calcofluor white stain

specimens in KOH are viewed under what magnification

100x amd 400x

Substance in lactophenol cotton blue that preserves fungi structures

lactic acid

killing agent in lactophenol cotton blue

phenol

stain in lactophenol cotton blue

aniline blue

can be added to 10% KOH or used alone for wet mount preparations of fungi

lactophenol coton blue

used for detection of C.neoformans in CSF through negative staining to democapsule

India Ink or Nigrosin

preferred method in temporary mount

calcofluor white stain

microscopy used in calcofluor white stain

fluorescence microscopy

fluoresce of organisms in calcofluor white stain

lur white or apple green

permanent mounts

periodic-acid schiff stain gram staun (hucker modficationj acid-fast stain (Kinyoun modification )Giemsa and Wright stain,papanicolaou stain ,Methenamine silver stain,Acridine orange,Auramine- rhodamine stain

stain fungal elements well;huphae of yeasts and molds can be readily distinguished

periodic acid schiff stain

carbohydratws present in fungus will stain - in periodic- acid schiff

purplish red

H& E slides may be restained with this method if fungus wall is invid

peripdic-acid schiff stain

all fungi are gram -/+

positive

fungi employs - in gram stain

crystal violet and ammonium oxalate

for detection of nocardia and B.dermatitidis

Acid fast stain

for detection of H.capsulatum and C.neoformans from blood or bone mattow specimens

Giemsa and Wright stain

cytotechnologist can detect fungal elements using this method

papanicolaou stain

for detection of fungi in hsitologic sections

methanamine silver stain

for itnew versicolor

acridine irange

flourescent of fungak elements in acridine orange

green

flourescent of epithelial cells in acrdine orange

orange

for nocardia

auramine-rhodamine stain

techniques for microscopic examination of fungal growth

Wet miunt,Tease Mount,Scotch or Cellophane tape preparation,Microslide culture

clsi document for antifungal susceptibility testing of yeasts

CLSI Document M27-A3

CLSI Document for antifungal susceptibility testing of moulds

CLSI Document M38-A2

CLSI DOCUMENT A Method for antifungal disk diffusion susceptibility testing of yeasts

Clsi dOcument M44-a2

clsi document a method for sntifungal disk diffusion susceptibility testing of nin-dermatophyte filamentius fungi

Clsi m51-A

angle of bent wire in wet mount

the portion in wet mount is placed in a slide and - is added

drop of lactophenol cotton or aniline blue is added

second needle in tease mount is used for

tease apart the fungus to form a thin kayer

added in tease mount for permanent mounting

10% PVA

Involves application of double-sticky tape or cellophane tape looped back on itself to the surface of the fungal colony

cellophane method

Optimal examination method for preservation of fungal morphology

microslide culture method

special tests

HAIR PERFORATION TEST  EXOANTIGEN TEST  RAPID UREASE TEST  RAPID NITRATE REDUCTASE TEST  LEVODOPA-FERRIC CITRATE TEST  GERM TUBE TEST  CARBOHYDRATE UTLIZATION TEST  COMMERCIAL YEAST IDENTIFICATION system

specimen used in hair perforation or baiting test

prepubertal hair

incubation period in hair perforst

25C for 10-14 days

antibodies developed against particular mycelial antigens will react specifically in a gel immunodiffusion precipitin test. The molds can be identified definitively by an antigen-antibody reaction

exoantigen test

Used in ghe microdiffusuon test of exoantigen

merthiolate extract of mature fungus

this is tested against positive control antiserum of exiantigen test

phenolized agar adjacent to the control antigen well

incubation in exoantigen test

25C

test that use either UREA-R broth or Urea Base

rapid urease test

incubation period in rapid urease test

4 hiurs at 37C

positive result color in Rapid urease test

pink to purple

Based on the ability yeast organism to produce nitrate reductase cell wall is dissociated with benzalkonium chloride to reduce nitrate to nitrite.

rapid nitrate reductase test

incubation period in rapid nitrate reductase test

45C for 10 minutes

reagents added in rapid nitrate reductase test

N-naphthylethylenediamine and sulfanilic acid

positive color in Rapid Nitrate reductase test

red

added in rapid nitrate reductase test if no color appears after 10 minites

pinch of zinc dust

also known as levodopa

ferric citrate test

C. neoformans is known to produce the enzyme phenol oxidase which when made to react with dihydroxyphenylalanine in the presence of ferric citrate forms a dark pigmented compound melanin

Ferric citrate test

incubation un ferric citrate test

37C and examined at 30-minute interval for up to 6 hours

specifically ysed to identify C.albicans

germ tube test

serun germ tube test incubatiin

35C for up to 3 hours

Yeast and yeast-like fungi are inoculated onto a carbohydrate-free medium  Carbohydrate-containing filter paper disks are added and utilization is determined by a color change or growth around the disk

carbohydrate utilization test

commercial yeast Id systems

API-20C yeast system  API Yeast-Ident system  Uni-Yeast Tek system  AutoMicrobic system  Abbott Quantum II system

incubation in yeast morphology test

25-30 for 24-72 hours

Document covers requirements for use of the broth microdilution method  Approved for the testing of Candida spp. and C.neoformans

CLSI Document M27-A3

standard microdilution method for molds that cause invasive and cutaneous infections

CLSI Document M38-A2

uses a dried colorimetric microdilution panel in a microtiter format for antifungal testing.

SENSITIRE YEASTINE