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Questions and Answers
What must be done before opening the autoclave to avoid explosion?
What must be done before opening the autoclave to avoid explosion?
- Increase the temperature rapidly
- Wait for pressure and temperature to lower (correct)
- Open the autoclave immediately
- Remove all contents from the autoclave
At what temperature should the agar be cooled to before dispensing?
At what temperature should the agar be cooled to before dispensing?
- 45 to 50°C (correct)
- 10 to 15°C
- 30 to 35°C
- 60 to 65°C
Which agar medium requires the use of an alcohol lamp for disinfection?
Which agar medium requires the use of an alcohol lamp for disinfection?
- Brain-heart infusion agar
- Littman oxgall agar
- Mycosel / mycobiotic agar
- Sabouraud Dextrose Agar (correct)
How much agar is required to prepare 1000mL of Sabouraud Dextrose Agar?
How much agar is required to prepare 1000mL of Sabouraud Dextrose Agar?
Which of the following media is specifically designed for Candida?
Which of the following media is specifically designed for Candida?
When preparing agar, what is the first step to take?
When preparing agar, what is the first step to take?
What type of agar medium is BHI agar classified as?
What type of agar medium is BHI agar classified as?
What is a crucial practice when working in a mycology laboratory?
What is a crucial practice when working in a mycology laboratory?
What is the purpose of using Periodic Acid-Schiff (PAS) stain?
What is the purpose of using Periodic Acid-Schiff (PAS) stain?
Which technique is NOT mentioned as a type of culture method?
Which technique is NOT mentioned as a type of culture method?
What should be done prior to using Wood’s lamp on a patient?
What should be done prior to using Wood’s lamp on a patient?
How far should Wood’s lamp be placed from the infected skin area?
How far should Wood’s lamp be placed from the infected skin area?
What is the recommended incubation period for most fungi detection?
What is the recommended incubation period for most fungi detection?
Which of the following is NOT a characteristic of the biphasic culture system?
Which of the following is NOT a characteristic of the biphasic culture system?
In what conditions should Wood’s lamp be used for optimal results?
In what conditions should Wood’s lamp be used for optimal results?
Which of the following is included in the final diagnosis when assessing skin, hair, and nails?
Which of the following is included in the final diagnosis when assessing skin, hair, and nails?
What temperature should Sabouraud's dextrose agar be incubated at for culture maintenance?
What temperature should Sabouraud's dextrose agar be incubated at for culture maintenance?
Which staining method is NOT mentioned for examining tissue sections?
Which staining method is NOT mentioned for examining tissue sections?
How long should cultures be maintained for Brain Heart Infusion Agar (BHIA)?
How long should cultures be maintained for Brain Heart Infusion Agar (BHIA)?
What agent is used as a mucolytic for viscous specimens?
What agent is used as a mucolytic for viscous specimens?
What is the initial step to process urine specimens for culture?
What is the initial step to process urine specimens for culture?
For direct microscopy examination of sediment, which solution is used to prepare the smear?
For direct microscopy examination of sediment, which solution is used to prepare the smear?
When inoculating viscous specimens, which type of swab is used?
When inoculating viscous specimens, which type of swab is used?
Which of the following media is NOT mentioned for culture maintenance?
Which of the following media is NOT mentioned for culture maintenance?
Which type of tinea infection is located on the scalp?
Which type of tinea infection is located on the scalp?
What type of medium is typically used for the initial isolation of pathogens and saprobes?
What type of medium is typically used for the initial isolation of pathogens and saprobes?
Which fungal infection is commonly associated with the groin area?
Which fungal infection is commonly associated with the groin area?
Which of the following types of tinea is known to affect the nails?
Which of the following types of tinea is known to affect the nails?
What is the main characteristic of dimorphic fungi when grown on SDA at 22°C?
What is the main characteristic of dimorphic fungi when grown on SDA at 22°C?
What is the growth rate for slow growers?
What is the growth rate for slow growers?
Which colony texture is characterized by irregular strands?
Which colony texture is characterized by irregular strands?
What does the elevation of a mold colony refer to?
What does the elevation of a mold colony refer to?
Which shape describes colonies that have a raised or bulging center?
Which shape describes colonies that have a raised or bulging center?
How long must fungi cultures be maintained before disposal?
How long must fungi cultures be maintained before disposal?
What does opacity of a colony indicate?
What does opacity of a colony indicate?
Which term describes colonies that are colored on the surface but have a different pigmentation underneath?
Which term describes colonies that are colored on the surface but have a different pigmentation underneath?
What characteristic of a colony helps microbiologists identify it?
What characteristic of a colony helps microbiologists identify it?
Which type of colony has a smooth margin?
Which type of colony has a smooth margin?
What type of colony is described as having a concave or 'innie' center?
What type of colony is described as having a concave or 'innie' center?
What is the characteristic of colonies that are opaque?
What is the characteristic of colonies that are opaque?
Which colony surface texture resembles cotton and has irregularities?
Which colony surface texture resembles cotton and has irregularities?
Which characteristic does not vary among species of the same genus?
Which characteristic does not vary among species of the same genus?
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Study Notes
Culture Media Used in Mycology Laboratory
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Various culture media for fungal growth:
- Birdseed Agar
- Brain-Heart Infusion Agar
- Canavanine-Glycine-Bromthymol Blue Agar
- Chromogenic Agar for Candida
- Cornmeal Agar
- Littman Oxgall Agar
- Mycosel / Mycobiotic Agar
- Potato Dextrose Agar/Broth
- Rice Grains Medium
- Sabouraud Dextrose Agar/Broth
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Autoclave Procedure:
- Start timing when pressure and temperature reach desired levels.
- Allow pressure and temperature to lower before opening to prevent explosion.
-
Preparation of Sabouraud Dextrose Agar (SDA):
- Weigh 65 grams of agar per 1000 mL of purified water, or 32.5 grams per 500 mL.
- Heat to boiling for dissolution.
Wood's Lamp Usage
- Clean skin to remove products before use.
- Wait 60 seconds after turning on.
- Use in a dark room, 10-30 cm from the skin.
- Patient should close their eyes during examination.
Primary Fungal Culture Medium
-
Expected growth outcomes at 22°C on SDA:
- Initial isolation of pathogens and saprobes.
- Dimorphic fungi may show mycelial phase.
-
Inhibitory Medium:
- SDA with antibiotics inhibits saprobes for accurate isolation.
Tissue Biopsies and Direct Microscopy
-
Essential stains for biopsy examination:
- Hematoxylin and Eosin (H&E)
- Grocott's Methenamine Silver (GMS)
- Periodic Acid-Schiff (PAS)
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Examination of cultured samples must occur after 4 weeks.
Urine and Respiratory Specimens
- For urine: Centrifuge sediment, prepare a direct smear in KOH.
- For respiratory specimens: Use Dacron swabs and N-acetyl-L-cysteine for inoculation.
Fungal Growth Rates
- Rapid growers: Appear within 1-3 days.
- Intermediate growers: Appear within 5-9 days.
- Slow growers: Appear up to 2 weeks.
Mold Colony Morphology
-
Colony characteristics aid in identifying species:
- Variability in color, shape, and growth form.
-
Elevation and Margin Types:
- Raised, flat, umbilicate, and umbonate elevations.
- Entire, undulate, and lobate margins.
Colonies' Surface Texture and Opacity
- Texture descriptions: Smooth, glistening, rough, wrinkled, cottony, velvety, granular.
- Opacity assessments: Transparent, translucent, opaque.
- Color: Surface and reverse pigmentation indicating type and health of the colony.
Common Fungal Locations and Types
- Tinea infections:
- Capitis: Scalp
- Faciei: Face
- Barbae: Beard
- Corporis: Body
- Manuum: Hands
- Ungulum: Nails
- Cruris: Groin
- Pedis: Feet
Identification Techniques
- Biphasic culture systems utilize both agar and broth for growth assessment.
- Culture incubations generally maintained for a minimum of 4 weeks before disposal.
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