This test is used to identify the bacteria (such as Klebsiella pneumonia) which are able to
hydrolyze esculin (a glycoside).

v Requirements:-

1.      Slant containing esculin

2.      24 hour broth culture

 v Procedure:-

1.      Inoculate the medium with one drop of 24 hour

2.      broth culture

3.      Examine the slants

v Observations:-

1.      Blackening of slant: Klebsiella pneumonia. (Positive test)

2.      No blackening of slant: Negative test

When observed for fluorescence under Wood’s lamp, or positive test, loss of fluorescence is
seen under Wood’s lamp. In the case of negative test there was no loss of flfluorescence.

v Quality control:-

1.       Use following microorganisms to confirm the reliability of reagents:

2.       Positive control: Klebsiella pneumonia

Negative test: Shigella flexneri