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House Microbiology > Laboratory Diagnosis Of Fungi > Flashcards

Laboratory Diagnosis Of Fungi Flashcards

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1
Q

Collection of specimens:

Skin specimens - clean with _________ to remove dirt, oil and surface saprophytes.

Nails - cleaned with ________. It is Usually ______; need to be _______ before inoculating to media.

Hair - obtained from _______________ ; hair can be obtained by ______,______ , or with a _______. A ________ can be helpful in locating infected areas.

Body fluids - normal sterile collection procedures.

A

70% alcohol

70% alcohol ; clipped; finely minced

edge of infected area of scalp; plucking, brushing; sticky tape; Wood’s lamp

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2
Q

Preparation of specimens for transport:

Hair & nails sent in a _________ , inside proper container.

Other specimens are usually sent _____ or ______

A

dry envelope

frozen; on dry ice.

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3
Q

Packaging of specimens - must meet _____________.

Cultures must be on _____ media (not ____).

Inside labeling information: ______,_____,_______
Outside labeling - _______________

A

biohazard regulations

tubed; plates

patient ID, specimen source, suspected organism.

WARNING: POTENTIAL PATHOGEN

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4
Q

Processing of specimen to recover fungus:
Body fluids -

CSF - should be ______ ; examine _____ _____scopically, then ________.

Pleural fluid, sputum, and bronchial aspiration - specimen must be _______ , as _____ would overgrow pathogens such as ________. The Specimens may be refrigerated up to _________

Gastric washings - same as for ______ fluids.

A

centrifuged; sediment; micro; inoculate media

fresh; saprophytes; H. capsulatum

2 hours.

pleural

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5
Q

Direct examination of specimens:

Direct exam is required on any biological material sent to lab for fungus culture.

Examine for ____,______,_____,____,____

A

spores, hyphae, mycelial elements, budding yeast, mycotic granules.

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6
Q

Direct examination of specimens:

Wet mount - good for ______; examination is done in _____ environment, so that ________ is minimal.

KOH - done on _____,_____,_____,_____,_____ specimens. KOH _____________ so fungal elements may be seen.

A

yeast

natural; loss of fragile structure

skin scrapings, hail, nails, sputum, vaginal

clears tissue cells

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7
Q

Processing of specimen to recover fungus:
Body fluids - (continued)

Genito-urinary specimens - _______ specimen preferred; should be _____ and _______ on media.

Blood/bone marrow - _________ to _____ and _____

Wound abscess or drainage - culture _____ if actinomycosis is suspected.

Tissue specimens - examine for _____,______, or ______; mince _____cally, inoculate on media.

A

first morning ; centrifuged & inoculated

inoculate to BHI broth and BHI slant.

anaerobically

pus, caseous material or granules

asepti

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8
Q

Staining

______________________ - very popular for quick evaluation of fungal structures; stains chitin in cell walls of fungi.

____________________ - stains polysaccharide in the cell wall of fungi.

_____________________ Stain - silver nitrate outlines fungi in black due to the silver precipitating on the fungi cell wall.

A

Lactophenol Cotton Blue

Periodic Acid - Schiff Stain (PAS)

Gomori Methenamine Silver

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9
Q

With Periodic Acid - Schiff Stain (PAS), Fungi stain ______ with ____ nuclei.

A

pink-red

blue

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10
Q

Staining
Stains used: (continued)

___________ Stain - extremely specific method of detecting fungi in tissues or fluids.
_________ Stain - good for initial differentiation of dimorphic fungi. Works well on sputum smears.

A

Fluorescent Antibody

Papanicolaou

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11
Q

In Gridley Stain - Hyphae and yeast stain _______ or ____ . Tissues stain ________ and background is ______.

A

dark blue or rose

deep blue ; yellow

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12
Q

Mayer Mucicarmine Stain - will stain capsules of Cryptococcus neoformans _________

A

deep rose.

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13
Q

Gram Stain - generally fungi are gram _______; but ______ and _____ are gram variable.

A

positive

Actinomyces and Nocardia

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14
Q

Staining
Stains used: (continued)

Acid-Fast Stain - used to differentiate the __________ from other ________

Giemsa Stain - used for _____ and ______ specimens.

India Ink - demonstrates the _______ of ________ in _____ specimens.

A

acid-fast Nocardia; aerobic Actinomyces.

blood and bone marrow

capsule of Cryptococcus neoformans; CSF

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15
Q

Fungal Culturing:
Culture
______ media is used rather than _____ media.

A

Tubed

plated

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16
Q

Fungal Culturing:
Culture
Tubed media is used rather than plated media.

Why

Less chance for _______
Less chance for ______.
Easier _______.

A

spore release into the environment.

dehydration

storage

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17
Q

The agar in a tube is inoculated in a ________.

Preliminary identification is based on __________ on various media.

A

straight line

differential growth patterns

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18
Q

Fungi culture media

__________ agar - classic medium, recommended for most studies.

A

Sabouraud’s dextrose

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19
Q

Sabouraud’s dextrose agar with ______ which inhibits bacterial growth

A

chloramphenicol

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20
Q

______ agar - a commercially produced agar containing chloramphenicol to inhibit bacterial growth, and _______ to inhibit saprophytic fungi and some yeasts (including C. neoformans).

A

Mycosel

cycloheximide

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21
Q

________ and _______ are opportunistic pathogens. Cycloheximide will prevent their growth. Bacteria-like fungi (such as _________ ) are inhibited by chloramphenicol.

A

Aspergillus and Scopulariopsis

Actinomycetes

22
Q

BHI- ______

A

Brain heart infusion

23
Q

Which is more enriched

BHI or Sab-Dex?

A

Brain heart infusion slant (BHI) is more enriched than Sab-Dex.

24
Q

BHI is used For recovery of H. capsulatum.

T/F

A

T

25
Q

______ and _________ agar are used in slide cultures to induce spore formation, which aids in identification.

A

Potato-dextrose agar (PDA) and Corn-meal

26
Q

Special media

Caffeic Acid Agar - Cryptococcus neoformans will produce ______ resulting in _____ colonies (must protect media from ______).

Birdseed Agar - used to _________ from contaminated cultures.

A

melanin ; black; light

isolate Cryptococcus neoformans

27
Q

Special media

KT Medium & Kelley Agar - used to convert dimorphic fungus ________ from _____ to _____ form.

A

Blastomyces dermatitidis

mycelial to yeast

28
Q

Special media

Modified Converse Liquid Medium (Levine’s) - used to promote ______ production by _________

A

spherule

Coccidioides immitis.

29
Q

Incubation

Fungal growth requirements -

Temperature -____ temperature (__-__C.) for most fungi.

Nocardia sp. and some dimorphic organisms grow best at ___ degrees C.

A

room; 25-30

37

30
Q

Any fungus capable of growing at 37 C, should be considered potentially __________.

A

pathogenic

31
Q

Incubation Atmosphere

True fungi is generally _______ but there are some ______ among the _____ fungi

A

Aerobes

Anaerobes; bacteria- like

32
Q

Incubation time:

Generally, cultures are held for ______

A

4 weeks

33
Q

Some yeasts grow overnight

T/F

A

T

34
Q

Saprophytes are slow growers( several days)

T/F

A

F

Fast growers (several days)

35
Q

Incubation time

Paracoccidioides brasiliensis may require ______ weeks

A

4-5

36
Q

Incubation time

____ weeks are recommended if Histoplasma capsulatum is suspected.

A

10

37
Q

Colony Morphology

Surface topography - some fungal colonies may be ____ growing, covering the _________; others may grow in a ______ manner.

A

free; entire surface of agar

restricted

38
Q

Colony morphology

Surface texture - cottony or wooly (floccose), granular, chalky, velvety, powdery, suede, silky, glabrous (smooth, creamy), or waxy.

T/F

A

T

39
Q

Colony Morphology (continued):
Pigmentation - Fungi may be colorless or brightly colored. Color may be on ______ itself, on its ______ apparatus, on the _____, or _______of the colony (reverse pigmentation).

A

fungus

sporulating

agar; on the bottom

40
Q

Mycelium -
Vegetative mycelium - provides _________.

Aerial mycelium –_________.

A

nutrition

reproductive

41
Q

Biochemical studies - used to ID yeast and yeast- like organisms.

Carbohydrate fermentation -
____________ of a carbohydrate under ———— conditions as determined by _____ and _____ production.

A

Growth and utilization

anaerobic

acid and gas

42
Q

Biochemical studies - used to ID yeast and yeast- like organisms.
Carbohydrate fermentation -

Specimen is inoculated ________.

__________ is the indicator.

Acid production turns _____ to ______.

Gas is detected by ____________ trapped in the fermentation tube.

Observe every _______ for ———-

A

beneath the broth

Bromcresol purple

purple to yellow.

appearance of bubbles

48 hours for 14 days.

43
Q

Biochemical test
Carbohydrate assimilation -

Determines ability to ______________

A

utilize a carbohydrate as sole source of carbon.

44
Q

Biochemical test
Carbohydrate assimilation -

Bromcresol purple indicator turns from _____ to ______

Tubes unchanged (as determined by __________) after _____ days are negative.

A

purple to yellow.

comparing to a blank tube; 10

45
Q

API;_________ for yeast identification

A

Auxocolor panels

46
Q

Biochemical studies - (continued)
Nitrogen assimilation -

Utilizes ____ tubes with differing sources of nitrogen.

___________ is the indicator

Color change of _____ to _____ is positive

A

3

Bromthymol blue

blue to yellow

47
Q

Growth on specific agars:

______________ agar - Urea is hydrolyzed by some yeast to form ammonia (pH increases) which turns media from _______ to _______

________ medium (must protect media from ____) - Production of melanin by Cryptococcus neoformans resulting in black colonies.

A

Christensen’s urea

yellow to dark pink.

Caffeic acid; light

48
Q

Germ tube -_______ and ______ produce germ tubes when incubated in a protein medium.

A

Candidia albicans & Candidia stellatoidea

49
Q

Demonstration of chlamydospores - Yeast is inoculated by jabbing appropriate agar (___________ with _______) and observed every 24 hours for 3 days for chlamydospore production.

A

Cornmeal with tween 80

50
Q

Serology
Hypersensitivity

Skin tests - demonstrates _________ to a fungus.

Serological tests - demonstrates __________ to a fungus; sera should be drawn in pairs (______ and _____).

A

T-cell immunity (cellular)

B-cell immunity (humoral)

acute and convalescent

51
Q

Serology
Hypersensitivity: ( _____ tests and _____ tests)

A

skin

serological

52
Q

Which is faster,

Culture or serology

A

Serology is Faster compared to culture

House Microbiology (93 decks)

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