Litmus Milk

Done by:
Alkaser40
 Purpose

Litmus milk is a milk-based medium used to distinguish between different

species of bacteria. The lactose (milk sugar), litmus (pH indicator), and casein
(milk protein) contained within the medium can all be metabolized by different
types of bacteria.
Since milk is usually the first substrate used to maintain bacteria, this test
allows for accurate depiction of bacterial types. The addition of litmus, other
than explaining the pH type, acts as an oxidation-reduction indicator. The test
itself tells whether the bacterium can ferment lactose, reduce litmus, form clots,
form gas, or start peptonization.
 What to observe?
• If lactose is fermented, the solution should turn pink. If gas is
produced during fermentation, you may be able to see bubble or
cracks in the milky medium (but this is often difficult to observe).
• If lactose is not fermented and proteins are instead used for energy,
the solution will become alkaline and more blue.
• Casein protein may be digested. This will coagulate the milk to form
a curd (a solid).
• The casein may be metabolized all the way down to individual
amino acids. This process, called peptonization, results in a clear
(not milky) liquid that is usually brown in color.
• The litmus may be reduced and become colorless. The culture will
then look milk white.
 Methods

1. Obtain two Litmus Milk broths from the back shelf.

2. Inoculate one broth with cells from your stock unknown culture.
Leave the other broth uninoculated (this will act as a control).
3. Incubate both broths at appropriate temperature (which ever is a
better growth temperature for your organism).
4. Observe your cultures over the next several days. Changes should
be visible in five to seven days.
 Results
Test name obsevation Diagram
after a week
Acid reduction Pink color

Alkaline Blue color

Control No change
 Results
Test name obsevation Diagram
after a week
Reduction Pink Top, White
Bottom,
somewhat solid
clot Milky color

Peptonization Blue top and

white bottom