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14.

Isolations and identification


of bacteria
Isolation techniques
 direct inoculation
 streaking
 enrichment cultures
identification and classification
 morphological characteristics
 cultural characteristics
biochemical tests
 carbohydrate dissimilation:
 sugar fermentation
 starch hydrolysis
 citrate utilization
 MRVP tests
 proteolytic tests:
 gelatin hydrolysis
 test for hydrogen sulfide
 indole test
 Fermentation and proteolysis of milk(action
on litmus milk) :
 acid clot
 stormy clot
 sweet clot
 peptonization
 Miscellaneous tests
 urease test
 catalase test
 coagulase test
 nitrate reduction
 oxidase test
Rapid identification systems [(eg: API
system)(Analytab products Inc.)]:
stiff plastic strip containing 20 ind. Microtubes, each
has a depression.
Each tube contains a dehydrated medium at the
bottom.
The dried media is reconstituted into aq form when
the inoculum of the test org is added.
The strip is then incubated
results are read by colour change using
charts/computer assisted identification
serological tests: studies blood serum and
immune responses that are evident in serum
eg: slide agglutination test: unknown bac drop into
several slides; different known antiserum is added to
each sample; bac agglutinate when mixed with
antibody (positive test)
eg: ELISA
eg: Western blotting
phage typing: determine which phage a bac is
susceptible to. Phages are bacterial viruses that can
cause lysis of the bacterial cells they infect. Food
associated infections can be traced by phage typing.
Eg: a)plate covered by bac
 b) a drop of different phage is placed on the bac.
 C) If phage infect the bac., the bac will lyse, clearings
appear.
nuclei acid hybridization: the procedure
assumes that if two species are similar or related, a
major portion of their nucleic acid sequence will also
be similar.
The process measures the ability of DNA strands
from one org. to hybridize with the DNA strands of
another org.
eg: southern blotting.
numerical taxonomy: a system that uses
overall degrees of similarity and large # of
characteristics to determine the taxonomic position of
an org.; allows org. of unknown affiliation to be
identified as members of established taxa; compared
by computer.