Lab 1 Diagnostic: Bacteriology Assistant Lecturer Hiba Ahmed

Diagnostic Bacteriology

Before one can proceed to identify a microorganism, the

characteristics of that organism have to be determined in details. The
major characteristics which are observed include the following:

A. Cultural Characteristics :-
In clinical terms include (shape, size, color, elevation and other
characteristics of the colony formed on the culture plate ).
In taxonomy includes :- nutrient requirements for the growth of the
organism and the physical factors such as temperature, pH and the
incubation period . These factors are used to identify certain
pathogenic species but less commonly used in routine procedures.

B. Morphology and Staining :-

This includes :- (microscopic appearance , size of the individual
cells, cell shape, arrangement and staining reaction if differential).
Example:-A gram stained film prepared from a pure culture of
certain microorganism shows the following:
-Small spherical cells "Cocci"
-Arranged in clusters
-Gram positive is violet in color and gram negative is pink color .
Some laboratories which have a little facility could give the report of
a microbiological examination of a clinical specimen just by stating
their morphological characteristics.
 Lab 1 Diagnostic: Bacteriology Assistant Lecturer Hiba Ahmed

C. Biochemical Characteristics :-
In general , biochemical tests are measuring the amount or activity of
a particular enzyme or protein, some important biochemical tests for
diagnosis :-
Coagulase , Catalase , Citrate , DNase , Hippurate , Hydrogen sulfide
production , Lecithinase ,Mixed acid fermentation, Oxidase, Urease ,
((Indol ,methyl red , Voges-Proskauer (VP) and Simmon's citrate test
and IMViC test )) etc.

D. Serological Characteristics :-
Sometimes, to identify a species as E. coli is insufficient, for the
reason that some strains of this organism are non-pathogenic and
others are highly associated with diseases. Serological testing in such
case will identify the exact strain number based on testing against
prepared specific antisera. In-Vivo serological tests (skin tests) are of
great value in the diagnosis of many bacterial, fungal and viral
infections.
 Lab 1 Diagnostic: Bacteriology Assistant Lecturer Hiba Ahmed

E. Other Characteristics :-
To identify some strains of bacteria, one may need to look for other
characteristics than those mentioned above. Phage typing and animal
inoculation are examples of uncommon techniques used in the
identification process.
Phage typing is a method used for detecting single strains of bacteria. It is used to trace
the source of outbreaks of infections. The viruses that infect bacteria are called
bacteriophages ("phages" for short) and some of these can only infect a single strain of
bacteria.

F. Molecular diagnostics :-
Molecular diagnostics is a collection of techniques used to analyses
biological markers in the genome and genetic code , for example
PCR techniques .

G. Automated systems for identification of microorganisms:-

(Vitek system)
There are currently four reagent cards available for the identification
of different organism classes as follows :-
1- GN :- gram negative (ferment and non-ferment bacilli )
2- GP :- gram positive cocci and non-spore forming bacilli
3- YST :- yeast and yeast like organisms
4- BCL :- gram positive spore-forming bacilli .

Types of culture media

 Lab 1 Diagnostic: Bacteriology Assistant Lecturer Hiba Ahmed

Media are of different types on consistency and chemical

composition.
A. On Consistency:
1.Solid Media. For (colony character , bacterial spread , isolation and
pure culture ).
2.Liquid Media. used for ( abandoned growth and activation isolates)
B. On Chemical Composition :
1.Routine Laboratory Media :- These are classified into six types:
(1) Basal media :- for growth ex:- Nutrient broth, nutrient agar and
peptone water .
(2) Enriched media:- usually by adding blood, serum or egg ex:-
( blood agar).
(3) Selective media :- allows the growth of bacteria without the other
group ex :- (MacConkey agar)
(4) Indicator media(differential media):- an indicator is included in
the medium ex:-(blood agar)
(5) Transport media :- used when specimen cannot be cultured soon
after collection ex:- Cary-Blair medium, Amies medium, Stuart
medium.
(6) Storage media. used for storing the bacteria for a long period of
time ex:- Egg saline medium
2. Synthetic Media. These are chemically defined media prepared
from pure chemical substances. It is used in research work.