Lab #13 Carbohydrate Utilization
Anabolism
All tests on Practical Exam –require explanation of YOUR results
TERMS
1. Anabolism-Dehydration ________________
A + B
Lab #13 Carbohydrate Utilization
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Catabolism, amylase, oxidative, fermentative
2.
Catabolism- Decomposition/hydrolysis, releases energy
AB
A + B
A. Amylase
B. Exoenzyme vs. endoenzyme
C. Oxidative
D. Fermentative
Lab #13 Carbohydrate Utilization
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Starch Plate
Starch Plate:
1.
Starch + beef extract + agar
2.
Amylase (_______enzyme) hydrolyzes
3.
Growth check
4.
Results
A. + for amylase if ____________________________
under/around colony
B. - if __________-starch still present
C. QC
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OF-Glucose
OF-Glucose
1. Glucose, peptone, indicators, & low amount agar
(semi-solid)
2. Growth check
3. Oxidative vs. fermentative vs. no glucose utilization.
Requires 2 tubes:
Demo set up
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OF-Glucose Tube Examples-Avail in Atlas
How would the tubes below be interpreted? Why?
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OF-Glucose, continued
A. Without oil: ______________________________
B. W/ oil: ___________________________________
C. Results:
i. Both tubes color change to yellow: ____________________
ii. Tube w/o oil only turns yellow (top or all): _____________
iii. No change: _______________________________________
4. Motility (+ or -)
5. Gas
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Flow Chart Available in Atlas
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TSI Slants
TSI-Triple Sugar Iron
1. Glucose, lactose, sucrose, peptone, Fe, indicators
2. Slant vs. butt
Demo & discuss set up, see p.13-4
3. Results MUST be read at 24 hours:
A. Bubbles/Cracks: ____________________
B. Black: _____________________ when S removed from
amino acids reacts w/Fe in slant
C. Carb utilization: Change from _______________________
(compare to unused tube)
i. See lab manual.
D. Record: Slant/Butt H2S +/- If gas present, _________
butt symbol.
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TSI Diagrams in Lab Procedure
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TSI Tube Diagrams
How would the tubes below be interpreted? Why?
Lab #13 Carbohydrate Utilization
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TSI Flow Chart Available
in Atlas
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TSI Problems & Special Situations
1. Some bacteria can’t utilize any of the sugars.
A. How does TSI support their growth?
i.
_______________ used as nutrient
B. What color does their tube turn? Why?
i.
Slant _____________ than at start – _________________,
releasing ___________ products & indicator goes
_____________________________.
2. Organisms that utilize glucose will cause the both the slant & butt to
turn yellow, even if it can’t use lactose & sucrose. If the slant is not
read at 24 hours, the slant will turn back to red. Why?
A. ________________ of glucose present. After glucose gone,
organism must ________________ to continue growth.
B. ______________metabolism releases __________ products &
_____________ again. Indicator ____________________.
Lab #13 Carbohydrate Utilization
Biochemical Tests: Indole Spot Test
Indole
1. Tests for presence of __________________________
ability of bacteria to ______________________________
2. Test from ____________________ agar
A. Positive (+): _________________color in 1 -3 minutes
B. Negative (-): _________________ change
Note:
•We will use a different reagent.
•Same reaction but, positive
color =_________________
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Biochemical Tests: Oxidase Spot Test
Oxidase
1. Tests for presence of ________________________________
2. Positive results indicate bacteria can use _____for energy
using an ____________________________
3. Test from nutrient agar
A. Positive (+): ______________color in 10-30 seconds
B. Negative (-): ___________________________________
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DrySlide Oxidase Test
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