• BLOOD AGAR

• COMPOSITION

• USES

• PRINCIPLE
OF HE MOLY S I S

• T Y P E S OF HE MOLY S I S
BLOOD AGAR (BA)
• ENRICHED MEDIUM USED TO CULTURE THOSE BACTERIA OR MICROBES THAT
DO NOT GROW EASILY(CALLED “FASTIDIOUS” )AS THEY DEMAND A SPECIAL,
NUTRITIONAL ENVIRONMENT AS COMPARED TO THE ROUTINE BACTERIA.
COMPOSITION OF BLOOD AGAR
• 0.5% Peptone
• 0.3% beef extract/yeast extract
• 1.5% agar
• 0.5% NaCl
• Distilled water
(Since Blood Agar is made from Nutrient Agar, above is the composition of Nutrient Agar)
• 5% Sheep Blood
• pH should be from 7.2 to 7.6 (7.4)
USES
• . Blood Agar is used to grow pathogens which are more difficult to grow such
as Haemophilus influenzae, Streptococcus pneumoniae and Neisseria species.
• Used to detect and differentiate haemolytic bacteria
P R I N C I P L E O F H E M O LY S I S
• Blood agar is a solid growth medium which has RBC'S in it .The medium is used to
detect different bacteria on basis of their enzymes which break apart RBC's in
different ways ,the degree to which RBC'S are hemolysed helps in distinguishing
different bacteria
 TYPES OF
H E M O LY S I S • alpha (α) hemolysis
• beta (β) hemolysis
• gamma (γ) hemolysis
• alpha prime or wide zone alpha
hemolysis.

To know the type of hemolysis, the blood agar plate must be held up to a
light source and observed with the light coming from behind (transmitted
light).
ALPHA H E M O LY S I S

• Partial lysis of the RBC

• produce a greenish-grey or brownish discoloration around the colony.
• Alpha hemolysis is due to the reduction of RBC hemoglobin to methemoglobin in
the medium surrounding the colony.
• Streptococcus pneumoniae ,streptococci Viridans
 B E TA - H E M O LY S I S

• complete lysis of RBCs

• colorless zone surrounding and under the colony.
• Main examples- Group A beta-hemolytic streptococci-
Streptococcus pyogenes and Group B Streptococcus agalactiace
• beta-hemolytic colonies are better observed when plates are
incubated in increased Co2 concentration.
• Staphylococcus aureus, Listeria monocytogenes, and Bacillus cereus.
G A M M A O R N O N - H E M O LY S I S

• NO HEMOLYSIS OF RBC'S
• NO CHANGE IN THE MEDIUM UNDER AND SURROUNDING THE COLONIES
• ENTEROCOCCUS FAECALIS , STAPHYLOCOCCUS SAPROPHYTICUS AND
STAPHYLOCOCCUS EPIDERMIDIS(CONS)
ALPHA PRIME OR WIDE ZONE
A L P H A H E M O LY S I S

• A small zone of intact erythrocytes immediately adjacent to bacterial colony

• With a zone of complete red-cell hemolysis surrounding the zone of intact
erythrocytes.
• This type of hemolysis may be confused with β-hemolysis
• The most reliable method of determining wide-zone alpha-hemolysis is the use
pour plates prepared with rabbit blood in heart infusion agar base
• Example-some strains of streptococcus causing mastitis.
TA R G E T H E M O LY S I S

• Clostridium perfringens - characteristic “double zone” hemolysis also known

as target hemolysis.
• resulting from a narrow zone of complete haemolysis caused by theta-toxin and a
much wider zone of incomplete haemolysis due to alpha toxin.