AFB.2

Question 1

1. HOT METHOD
   - primary stain: carbol fuchsin conc.
   - mordant
   - acid alcohol conc.
   - counterstain
2. COLD METHOD
   - primary stain: carbol fuchsin conc.
   - mordant
   - acid alcohol conc.
   - counterstain

Answer 1

1. ZIEHL-NEELSEN
   - 3% phenol
   - heat
   - 3% hydrochloric acid + 5% ethanol
   - methylene blue
2. KINYOUNS METHOD
   - 9% phenol
   - tergitol
   - 3% sulfuric acid + 95% ethanol
   - methylene blue

Question 2

AFB SEEN (______ magnification)

• No AFB seen =
• Indeterminate (Request again) =
• 1+ =
• 2+ =
• 3+ =
• 4+ =

Answer 2

1000x

• 0
• 1-2 / 300 fields
• 1-9 / 100 fields
• 1-9 / 10 fields
• 1-9 / field
• > 9 / field

Question 3

Culture Media General Formulations

1. used for observing colony morphology, susceptibility testing
   - contains ____, ____, _____, _____,____, _____, _____ (neutralizes inhibitory and toxic effects of FA)
2. w/ malachite green (inhibits gram positive bacteria)
   - contains ____,_____,______
3. support proliferation of small inocula

Answer 3

1. Semisynthetic Agar Media
   - defined salts, vitamins, cofactors, oleic acid, catalase, glycerol, albumin
2. Inspissated Egg Media
   - defined salts, glycerol, complex organic substances
3. Broth Media

Question 4

BIOCHEMICAL RXN
1. Growth Rate
- rapid growers =
- exhibits false-positive growth
2. Pigment Production
- yellow to orange to red in light
- yellow to orange to red in dark & light
- do not develop pigment; not affected by light
3. Niacin Accumulation
-_____ plays important role that corpus during mycobacteria metabolism
- NO CONVERSION of niacin to
- they lack ______
- POSITIVE
- although all spp. produce nicotinic acid, ______ accumulates the largest amount

Answer 4

1. Growth Rate
   - 3-4 days
   - M.flavescens
2. Pigment Production
   - photochromogens
   - scotochromogens
   - nonchromogens
3. Niacin Accumulation
   -niacin (nicotinic acid)
   - niacin ribonucleotide
   - niacin-binding enzyme
   - accumulation of niacin imparted by yellow color
   - M. tuberculosis

Question 5

BIOCHEMICAL RXN
1. Nitrate Reduction
- possesses enzyme
- uses broth = _____ + _______
- or uses _____
- positive
2. Catalase &
Heat Stable
Catalase
- All are Catalase positive but Not all Heat Stable: negative for all ________
- Heat at ____C for ____mins
- reacted with _____
- positive

Answer 5

1. Nitrate Reduction
   - nitroreductase
   - sodium nitrate broth + hydrochloric acid, sulfanilamide, &
   N-l-naphthylethylenediamine
   - commercial filter paper strip
   - pink or red color
2. Catalase & Heat Stable
   - M.tuberculosis complex
   - 68C for 20mins
   - 30% hydrogen peroxide
   - foam bubbles

Question 6

BIOCHEM RXN

Tween 80 Hydrolysis
1. slow-growing scotochromogens & nonphotochromogens produce a _____ that can hydrolyze Tween 80
2. Tween 80 is a detergent ______
3. Tween 80 hydrolyzes into ____ & ______ (whereas pathogenic spp do not)
4. Indicator
5. Differentiates _______ (+) & _______ (-)

Answer 6

1. LIPASE
2. polyoxyethylene sorbitan monooleate
3. oleic acid & poly-oxyethylated sorbitol
4. neutral red
5. M.gordonae (Tween 80 +) from M.scrofulaceum (Tween 80 -)

Question 7

BIOCHEM TEST

1. Arylsulfatase
   - rate at which the enzyme breaks down ______ into _______
   - POSITIVE = _____ (presence of ______)
2. Pyrazinamidase
   - converts this enzyme to _____ & _______
   - when the latter is mixed with ______ it produces red color (+)

Answer 7

1. Arylsulfatase
   - tripotassium phenolphthalein into phenolphthalein
   - pink (sodium bicarbonate)
2. Pyrazinamidase
   - pyrazinoic acid & ammonia
   - ferric sulfate

Question 8

BIOCHEM TEST

1. Urease
   - converts urea to _____ and _____
   - positive
2. Growth in 5% NaCl
   - most mycobacteria cannot grow in 5% sodium chloride except _______
3. Iron Uptake
   - only the rapid growers are able to grow in _______
   - grows in ______
   - positive

Answer 8

1. Urease
   - ammonia & carbon dioxide
   - pink
2. Growth in 5% NaCl
   - M.triviale
3. Iron Uptake
   - 20% ferric citrate
   - egg-based slant
   - rusty brown