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Microbiology Lab exercise 14 -Fermentation

front 1

Fermentation produces

back 1

acids
alcohols
gas

front 2

True or false. Nutrient agar there is no differentiation.

back 2

True

front 3

Carbohydrate fermentation procedure

back 3

Obtain 4 of each carbohydrate fermentation tubes:
glucose
sucrose
lactose

Obtain one each of the following: E. coli, E. aerogenes, A. faecalis, P. vulgaris
inoculate each set with each bacteria

front 4

Why is the durham tube upside down?

back 4

to detect gas production

front 5

carbohydrate fermentation media:

back 5

broths
uninoculated color: red
contains:
one peptone
durham tube
phenol red (pH indicator)

front 6

Phenol red is a pH indicator

back 6

yellow- positive for sugar fermentation pH lower
red- no change pH stayed same
pink/magenta- it couldn't metabolize the sugar and use for food so it metabolized the protein pH higher

front 7

The carbo. tube is full and no bubbles

back 7

positive for acid fermentation

front 8

Carbo tube has bubbles in durham tube

back 8

positive for gas fermentation

front 9

carbo tube has no change in color or gas production

back 9

negative for fermentation

front 10

Carbo tube is orange

back 10

Intermediate zone- (-) for fermentation

front 11

True or false. Bacteria can ferment a variety of carbohydrates through different pathways. No single bacterial species is capable of them all.

back 11

true

front 12

_______ it typically the starting point of carbohydrate fermentation.

back 12

pyruvic acid

front 13

True or false. Acids and/or gases may be produced as products of fermentation.

back 13

true

front 14

Fermentation ability depends on ______ and the ______ it produces.

back 14

bacteria
enzymes

front 15

Carbohydrate fermentation theory
positive for acid production

back 15

pH decreases
phenol red becomes yellow and turns media yellow

front 16

If positive for gas production in carbohydrate fermentation

back 16

media in durham tube is displaced
bubble forms within durham tube

front 17

Carbohydrate fermentation results:
yellow broth bubble in durham tube

back 17

positive for fermentation with gas and acid products

front 18

Carbohydrate fermentation results:
yellow broth no bubble in tube

back 18

positive for fermentation with acid products

front 19

Carbohydrate fermentation results:
red broth no bubble in tubes

back 19

no fermentation- no acid or gas production

front 20

Carbohydrate fermentation results:
pink broth no bubble in tube

back 20

protein catabolism: alkaline end product
no acid or gas

front 21

Carbohydrate fermentation results:
orange broth

back 21

negative for fermentation no gas or acid

front 22

MRVP results

back 22

red is positive for both tests

front 23

MRVP is _____ test and they are _____ and ______.

back 23

2
methol red
vogues proskauer

front 24

The methol red test is testing for?

back 24

mixed stable acids fermentation
it overcomes the buffer and changes the pH

front 25

The VP test is for?

back 25

to check for the production of acetoin

front 26

Methyl red test you add __ drops to the MRVP broth

back 26

5

front 27

In the methyl red test if the broth remains red than it is _____________.

back 27

positive for the production of mixed stable acids

front 28

The MRVP test contains:

back 28

glucose
protein
phosphate buffer (resists pH change)

front 29

In the methyl red test it is used to detect large amounts of ________.

back 29

mixed stable acids
acids are stable and overcome buffer
cause pH change

front 30

True or false. In the methyl red test the only true indication of a positive fermentation for mixed stable acids is ______.

back 30

red color

front 31

The VP test you add
12 drops of reagent 1 which is _________.
And 2-3 drops of reagent 2 which is __________.

back 31

a-napthol
KOH

front 32

In VP test after adding reagents put paraffin cover and shake vigorously and let stand for 30 mins
Results:

back 32

Light brown tea color (-) for acetoin production
Red or red band at top is positive for acetoin production

front 33

In the VP test it is the fermentation of ______ that produces acetoin.

back 33

glucose

front 34

Citrate test is done in a _______ and you ______ to inoculate the tube.

back 34

slant-green (solid) and you zig zag up to inoculate it

front 35

Citrate media is:

back 35

slant-solid media/green
One carbon source- sodium citrate (carbon)
One nutrient source monoammonium (nutrient)
Bromthymol blue- pH indicator
dipotassium phosphate
magnesium sulfate (cofactor for enzymes)
agar

front 36

The positive result of citrate is

back 36

green-----> blue

front 37

For the citrate test the bacteria need both ____ and ______.

back 37

carbon and energy

front 38

The citrate test tests the bacteria's ability to

back 38

obtain both carbon and energy

front 39

The citrate test allows differentiation between different members of Enterobacteriacea

back 39

enteric bacteria

front 40

If the citrate test is positive bacteria

back 40

produce citrate lyase
make ammonia from ammonium
increase the pH of agar
media turns a deep Prussian blue

front 41

Citrate test results:
blue even a small amount

back 41

positive
citrate is utilized

front 42

Citrate test results:
No color change; growth

back 42

citrate is utilized; positive

front 43

Citrate test results:
no color no growth

back 43

citrate not utilized
negative citrate test

front 44

KOH is

back 44

potassium hydroxide

front 45

a-napthol is used with KOH to detect

back 45

acetoin