Blood Agar

• Used for cultivation of fastidious organisms and

studying haemolytic reactions.
• It provides improved and enhanced haemolysis.
• Addition of blood makes the medium more nutritious
by providing additional growth factors required by
fastidious organisms.
• Haemolytic reactions depend on the animal blood
used
 Composition

Ingredients Grams/Litre
Casein enzymic hydrolysate 14.000
Peptic digest of animal tissue 4.500
Yeast extract 4.500
Sodium chloride 5.000
Agar 12.500
Sheep Blood 5.000
Final pH [at 25° C ] 7.3±2
• Casein enzymic hydrolysate and yeast extract provide nitrogen, carbon, amino
acids and vitamins.

• Peptic digest of animal tissue is the nitrogen source.

• Sodium chloride maintains the osmotic balance.

 Principle

 Haemolysins are exotoxins produced by bacteria that lyse red blood

cells.
 The haemolytic reaction can be visualized on blood agar plates.
 On blood agar plates colonies of haemolytic bacteria may be
surrounded by clear, colourless zone where the red blood cells have
been lysed and the haemoglobin destroyed to a colourless compound.
 There are three types of hemolysis, designated alpha, beta and gamma
 Alpha hemolysis is a greenish
discoloration that surrounds a
bacterial colony growing on the
agar.
 This type of hemolysis
represents a partial
decomposition of the
hemoglobin of the red blood
cells.
 Alpha hemolysis is characteristic
of Streptococcus pneumonia.
 Beta hemolysis represents
a complete breakdown of the
hemoglobin of the red blood
cells in the vicinity of a
bacterial colony.
 There is a clearing of the
agar around a colony.
 Beta hemolysis is
characteristic of
Streptococcus pyogenes and
some strains of
Staphylococcus aureus.
 Gamma hemolysis is a lack
of hemolysis in the area
around a bacterial colony.
 A blood agar plate
displaying gamma
hemolysis actually appears
brownish.
 This is a normal reaction
of the blood to the growth
conditions used (37° C in
the presence of carbon
dioxide).
 Gamma hemolysis is a
characteristic
of Enterococcus faecalis.
 Limitations

 Sheep blood gives best results for Group A Streptococci.

 But sheep blood fails to support growth of Haemophilus
haemolyticus since sheep blood is deficient in pyridine
nucleotides.
 However when horse blood is used H. haemolyticus
colonies produce haemolysis and mimic Streptococcus
pyogenes .
 References

 Hi Media Labs
 https://www.encyclopedia.com/science/encyclopedi
as-almanacs-transcripts-and-maps/blood-agar-
hemolysis-and-hemolytic-reactions