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MIO Method

MIO medium is used to differentiate Enterobacteriaceae by testing for motility, ornithine decarboxylase activity, and indole production in a single tube. It contains nutrients, dextrose, bromocresol purple pH indicator, and low concentration agar. Motility is shown by diffuse growth away from the inoculation line. Ornithine decarboxylase activity alkalinizes the medium from acid pH caused by dextrose fermentation, turning it purple. Lack of the enzyme leaves it yellow. Indole production is detected by formation of red color in reagent layered on top of the medium.

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Maryam Adel
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249 views9 pages

MIO Method

MIO medium is used to differentiate Enterobacteriaceae by testing for motility, ornithine decarboxylase activity, and indole production in a single tube. It contains nutrients, dextrose, bromocresol purple pH indicator, and low concentration agar. Motility is shown by diffuse growth away from the inoculation line. Ornithine decarboxylase activity alkalinizes the medium from acid pH caused by dextrose fermentation, turning it purple. Lack of the enzyme leaves it yellow. Indole production is detected by formation of red color in reagent layered on top of the medium.

Uploaded by

Maryam Adel
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© © All Rights Reserved
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MOTILITY

INDOLE
Ornithine
(MIO)
medium
FORMULA
Gram weight per liter: 31.0gm/L
Pancreatic Digest of Gelatin 10.0gm
Pancreatic Digest of Casein 10.0gm
L-Ornithine 5.0gm
Yeast Extract 3.0gm
Dextrose 1.0gm ( all enterobacteriacae ferment dextrose)
Bromcresol Purple (indicator) 0.02gm
Agar 2.0gm
MIO Medium (Motility-Indole-Ornithine) is a semisolid medium used for the
differentiation of the Enterobacteriaceae group by motility, ornithine
decarboxylase activity and indole production (3 tests in one tube ).
Principles of the Procedure

 If the organisms possesses ornithine decarboxylase, it will be


activated in an acid environment created by the initial fermentation of
dextrose.(Stabbing center of the medium by inoculating needle )
 Once the amino acid is decarboxylated, diamine putescine is
produced. The result is an alkalinization of the medium, which turns
it purple.
 Organisms without the the enzyme, will remain acidic due to the
fermentation, resulting in a yellow color in the medium.

 Bromocresol purple is a pH indicator to indicate decarboxylase


activity; the low concentration of Bacteriological agar is for motility.
•Bacterial motility is evident by a
diffuse zone of growth extending out
from the line of inoculation. Some
organisms grow throughout the
entire medium, whereas others show
small areas or nodules that grow out
from the line of inoculation.
•The non-motile bacteria will only
grow in the soft agar tube  and only
the area where they are inoculated.

 .(Stabbing center of the medium by inoculating


needle )
A positive indole test is indicated by the formation of a
red color in the reagent layer on top of the agar deep
within seconds of adding the reagent.
(inoculation top of media by inoculating loop)
+Motility
-indole
+ornithine

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