LABORATORY EVALUATION (Macroscopic) Flashcards
Media used for the cultivation of asporogenous yeasts
Acetate ascospore agar
This formulation is a better sporulation medium than sodium acetate
potassium acetate
stain used in acetate ascospore agar
Kinyoun carbol-fuchsin acid-fast stain
What species is seen in acetate ascospore agar?
Saccharomyces cerevisiae
Medio used for the isolation of Cryptococcus spp. esp. C. neoformans and C. gattii which is unique in that they produce the enzyme phenol oxidase
Birdseed agar (syn. Niger seed agar)
true/false
Breakdown of substrate (Guizotia abyssinica seed or niger seeds) produces MELANIN which is absorbed into the yeast wall and imparts a tan to brown pigmentation of the colonies
true
reagent used in birdseed agar
chloramphenicol and creatinine
a selective agent that inhibits bacteria and some fungi
chloramphenicol
Enhances melanization of some strains of C. neoformans
creatinine
mediu used for the isolation and differentiation of candida spp.
Bismuth sulfite-glucose-glycine yeast (BiGGY) agar
true/false
Candida spp. reduce the bismuth sulfide (also acts as an inhibitor of bacterial growth) to bismuth sulfite which results in pigmentation
false
Candida spp. reduce the bismuth sulfite (also acts as an inhibitor of bacterial growth) to bismuth sulfide which results in pigmentation
nutritive bases of BiGGY
Peptone
Glucose
Yeast Extract
C. albicans vs C. tropicalis
in terms of color of colonies
C. albicans: Brown to balck colonies with no pigment and no sheen
C. tropicalis: Dark brown colonies with black centers, black pigment diffusion, and sheen
medium used for the cultivation and isolation of all fungi
Brain heart infusion agar (fungal formulation)
true/false
BHI contains 5% sheep red cell
false
10% kase
reagent used in BHI
chloramphenicol
gentamicin
cycloheximide
Inhibits overgrowth of saprophytic fungi
cycloheximide
An adaptation of birdseed agar.
Caffeic agar
what species is detected in caffeic agar?
C. neoformans, brown in color
medium used to distinguish C. neoformans from C. gattii
Canavanine-glycine-bromthymol blue agar
C. gattii vs C. neoformans C. neoformans var. neoformans
in terms of serotypes
C. gattii: serotype B and C
C. neoformans: serotype A
C. neoformans var. Neoformans: Serotype D
C. gattii vs C. neoformans C. neoformans var. neoformans
in terms of colonial color
C. gattii: cobalt blue
C. neoformans and C. neoformans var. Neoformans: greenish yellow
medium used for the isolation of clinically important yeasts
CHROMagar (BD BBL; BD Diagnostics Systems)
reagent used in CHROMagar
PEPTONE
GLUCOSE
CHLORAMPHENICOL
species detected in CHROMagar
C. krusei
C. glabrata
true/false
CHROMagar is more sensitive than SDA and helpful in identifying mixed cultures of yeasts, and it may enhance the rapid assimilation of trehalose by C. glabrata.
true
true/false
The medium is available with or without fluconazole, providing additional selection of fluconazole resistant viz., C. krusei
true
medium used in the presumptive identification of Cryptococcus, Trichosporon, Rhodotorula spp
Christensen’s urea agar
It uses urea hydrolysis which facilitates the separation of certain dermatophytes viz., T. mentagrophytes and T. rubrum
christensen’s urea agar
reagent used in christensen’s urea agar
UREA
phenol red
used for the cultivation and differentiation of T. mentagrophytes from T. rubrum on the basis of pigment production
Cornmeal agar with 1% dextrose
Used for the differentiation of Candida species on the basis of morphological characteristics.
Cornmeal agar with Tween 80
Used as the surfactant which is specifically incorporated in lieu of dextrose for the demonstration of pseudohyphal, chlamydospores, and arthrospores formation
Tween 80
Method used in for chlamydospore production wherein it is obtained by subsurface inoculation, or by placing a cover slip over the yeast inoculum, creating a microaerophilic environment
Dalmau method
used for the differentiation of Aspergillus spp
Czapek-Dox agar
These are the sole carbon and nitrogen sources in Czapek-Dox agar
Sucrose and sodium nitrate
Any bacteria or fungi that can use sodium nitrate as a nitrogen source can grow on this medium.
Czapek-Dox agar
For the recovery, selection, and differentiation of dermatophytes from keratinous specimens
Dermatophyte test medium (DTM
reagents used in DTM
Cycloheximide
Chloramphenicol
Phenol red indicator
true/false
Medium is red and turns yellow with growth of dermatophyte.
false!
baliktad and color beh
For the isolation and growth of lipo dependent Malassezia spp
Leeming and Notman medium
composition of Leeming and Notman medium
Ox bile
Glycerol monostearate
Glycerol
Tween 80
Cow’s milk (whole fat)
Medium may serve as an alternative to SDA because not all species can grow in this medium
Leeming and Notman medium
These are species of malassezia that cannot grow in Leeming and Notman medium
M. globosa, M. restricta, M. obtusa
For the isolation of fungi from contaminated specimen.
Littman oxgall agar
the selective agents inhibiting bacteria in littman oxgall agar
Crystal violet and streptomycin
This restricts the spreading of fungal colonies.
oxgall
For the isolation of dermatophytes but also for the isolation of other pathogenic fungi from specimens contaminated with saprophytic fungi and bacteria
Mycobiotic or Mycosel agar
Stimulate conidium production by fungi and stimulates pigment production in some dermatophytes
Potato Dextrose
used with the slide culture technique to view morphological characteristics
Potato Dextrose
Incorporation of this in Potato Dextrose medium lowers the pH, thereby inhibiting bacterial growth.
tartaric acid
Used for the selective cultivation of yeasts, molds and aciduric bacteria
Sabouraud dextrose agar (SDA)
reagents in SDA
Pancreatic digest of casein, peptic digest of animal tissue, dextrose at 4% conc. Buffered to a ph of 5.6
- Chloramphenicol
- Cycloheximide
- Gentamicin
- Ciprofloxacin
- Penicillin and (or) Streptomycin
What did Emmons do to modify SDA?
modified the original formulation by reducing the dextrose conc’n. To 2% and adjusting the pH nearly to neutrality 6.9 to 7.0
Petri dish vs Test tube
in terms of surface area
Petri dish: large (7,500 mm2)
Test tube: small (1,500 mm2)
Petri dish vs Test tube
in terms of oxygen supply
Petri dish: good
Test tube: poor
Petri dish vs Test tube
in terms of rate of drying
Petri dish: relatively fast
Test tube: relatively slow
Petri dish vs Test tube
in terms of security of closure
Petri dish: poor (lid is easily displaced)
Test tube: good
Petri dish vs Test tube
in terms of probability of dissemination
Petri dish: relatively large
Test tube: relatively small
Petri dish vs Test tube
in terms of detection of mixed culture
Petri dish: relatively easy
Test tube: relatively difficult
incubation of fungal culture of Mold forms, most common temperature for incubation
25 C to 30 C
incubation of fungal culture of Opportunistic and dimorphic organisms
30 C
incubation of fungal culture of Conversion to yeast phase for dimorphic fungi
25 C to 37 C
growth rate: <5 days
rapid growers
growth rate: 8-10 days
intermediate growers
growth rate: >11 days
slow growers
texture appears leather-like, or waxy little mycelium, seems to merge with the aga
Glabrous
texture Resembles plush or suede, short aerial hyphae of equal length
Velvety
texture Resembles colonies of other Staphylococcus spp. (formerly CoNS), “bacteria like” but
more dry and dull (waxy-pasty). No aerial mycelium, with a delicate fringe around the
colonies in BAP
Yeast like
texture Wooly or “Floccose”, large quantities of long aerial hyphae that becomes entangled and
may fill the entire petri dish
Cottony
texture Powdery due to heavy conidiation or sporulation, has even hyphae and abundant
conidia
Granular
topography: Presence of radial groves from the center of the culture toward the rim
Rugose
topography: Random folds (long, short, parallel at right angles or combination)
folded
topography: Have central depression (concavity) surrounded by raised edges. Resembles S.
pneumoniae colonies
Crateriform
topography: Have many warts or rough knobs on the surface
verrucose
topography: Brain-like convolutions
cerebriform
methods used in microscopic evaluation of growth in culture
tease mount
dalmau method
slide culture
scotch tape method
Method of preservation of culture where conidia and spores form a fresh culture are washed off in sterile water and placed in labelled vials
Storage in water
Method of preservation of culture: Labelling glass tubes with a screw-capped and placed in a freezer (-70oC)
Freezing
Method of preservation of culture: Layering an entire slant with mineral oil, capping the tube tightly and storing at room
temperature
Mineral oil overlays
Method of preservation of culture: Freeze drying with the use of special equipment (lyophilizer)
Lyophilization
methods in Immunologic identification of fungi
Immunodiffusion (ID)
Countercurrent immunoelectrophoresis (CIE)
Enzyme-linked immunosorbent assay (ELISA)
Complement-fixation test (CFTs)
Fluorescent-enzyme immunoassay (FEIA)
Found in the blood of patients with invasive fungal infections, and its ability to activate factor G of
the horse-shoe crab coagulation pathways that allows it to be measured and quantified.
1,3-β-D-glucan
this method of immunologic identification has the ability to detect organisms that are present in small numbers of that cannot be cultured.
Nucleic acid testing
this method of immunologic identification offers the potential to identify pathogens in biopsy samples that have been formalin-fixed and even wax-embedded
Nucleic acid testing
this method of immunologic identification offers the potential to identify pathogens in biopsy samples that have been formalin-fixed and even wax-embedded
Nucleic acid testing
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