Microbio Pracs Reviewer Bacteriology 2017
Microbio Pracs Reviewer Bacteriology 2017
Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Procedure:
1. Flood smeared area with CRYSTAL VIOLET stain
(initial stain) for 1 minute
2. Pour off stain, wash with gentle flow of water
3. Apply GRAM’s IODINE (mordant) for 1 minute, then
rinse off with water
4. Decolorize using ACETONE-ALCOHOL solution or
95% ETHYL ALCOHOL for 20 seconds
5. Apply SAFRANIN (counter stain) for 30 seconds, then
wash off with water
Report as follows:
Reporting Acid Fast bacilli in Sputum specimen
No AFB in at least 100 0 or (-)
fields
1-9 AFB in 100 fields +n (report actual AFB
count), Suggest repeat
collection
10-99 AFB in 100 fields 1+
1-10 AFB in at least 50 2+
Bacterium Background
fields
>10 AFB/field in at least 3+ Pink Blue
20 fields
Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Differentiated by
hemolysis, mannitol salt
agar growth, and
coagulase test
S. aureus
Deep gold to pale cream
or white colonies
Clear zone of hemolysis
S.epidermidis
Porcelain white colonies
Non-hemolytic
Credits to K. Erguiza SecB 2014 for the picture
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(+) Staphylococcus
(-) Streptococcus
Credits to K. Erguiza SecB 2014 for the picture
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MANNITOL
FERMENTATION Test
1. Streak on the Mannitol
Salt Agar plate
2. Observe color of colonies
▪ Yellow colonies =
fermentation of mannitol
3. Wrap plate and incubate
for 18-24 hours
COAGULASE TEST
I. Slide Method
1. Place 2 drops of sterile saline on slide
2. Pick a colony using loop, mix with the saline
3. Add 1 drop citrated fresh normal plasma
4. CLUMPING = positive!
(+) S. aureus
(-) S. epidermidis
COAGULASE TEST
II. Tube Method
1. Get tube containing 0.5 mL
citrated fresh plasma
2. Suspend colonies on tube
3. Incubate for 37 deg C in water
bath and read results at 4 hours
and 24 hours if still negative
4. COAGULATION (by tilting tube) =
positive!
5. Often, a clot will stick to the
bottom of the tube even if it is
inverted
(+) S. aureus
(-) S. epidermidis Credits to K. Erguiza SecB 2014 for the picture
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Streptococcus pneumoniae
Group B, Alpha hemolytic,
lancet-shaped, susceptible to
optochin in chocolate agar
plate (CAP)
POSITIVE QUELLUNG
REACTION
Viridans streptococci
Alpha hemolytic
Credits to K. Erguiza SecB 2014 for the picture
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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CATALASE TEST
1. Place a colony on a clean glass slide
2. Add a drop of 3% H2O2
3. Observe for bubble formation
OPTOCHIN TEST
Susceptible: Group B Strep i.e. S. pneumoniae
Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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I. NON-SPOREFORMER BACILLI
Corynebacterium diphtheria
Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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EMBA (Eosin
Methylene Blue
Agar)
Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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MacConkey Agar
Lactose fermenter
– large, red colonies
surrounded by
turbid zones
Nonlactose
fermenter –
colorless and
translucent colonies
Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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MacConkey Agar
Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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MacConkey Agar
Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Biochemical
Tests Procedure
Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Acid slant yellow Alkaline slant red Alkaline slant red Blackening of Cracks, bubbles
Acid butt yellow Acid butt yellow Alkaline butt red butt or displacement
of medium
All sugars GLUCOSE No sugar Produce H2S Produce gas
fermented fermented fermented (aerogenic)
Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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E. UREASE TEST
▪ Detects ability to split urea
to ammonia (basic!) and CO2
by the enzyme urease
Indicator: Phenol red
Medium: Urea Broth
▪ Test: Urease Test
E. UREASE TEST
Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Selective for
S. typhi
Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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A. Water Collection
1. Clean the mouth of faucet with 70% EtOH
2. Collect 100 mL of chlorinated (tap water) by
allowing water to flow about 1-5 minutes
3. Alternative: unchlorinated (deep well) water
4. Refrigerate while waiting to transport
5. Transport in ice (4 degrees Celsius)
Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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Computation of MPN
(Most Probable Number)
Sources: Notes from UST-FMS Micro Lab Manual, Pictures from UST-FMS 2017 Sec D & Internet
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