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Exercise No. 24:

Bacillus: Morphology & Cultural Characteristics
A. Direct Detection method
1. Gram Stain
Gram stain of Bacillus anthracis
Smear shows long gram positive bacilli with
square ends arranged in chains
Characteristic shape and arrangement of
Bacillus anthracis likens the microbe to
bamboo rods

Gram stain of Bacillus cereus

Smear shows gram positive bacilli arranged
in chains

2. Spore Stain
Spore stain of Bacillus anthracis
Spores are oval, located centrally to
subterminally, and usually not swollen

B. Cultivation
1. Colony Characteristics
A. Blood Agar Medium
Blood agar plate culture of Bacillus anthracis
Heavy growth of non-hemolytic large, flat and
irregular, 4 to 5 mm in diameter, with slight
undulate margin, granular, coarse colonies

Colony consistency of Bacillus anthracis is

likened to a beaten egg white
When lifted by an inoculating loop, colonies
show a tenacity that allows them to be pulled up
and stay upright.

Dissecting microscopy of Bacillus anthracis

colonies
Shows numerous undulated outgrowths
consisting of long filamentous chains of bacilli
consistent with the characteristic colonial
swirling projections (medusa head appearance)

Blood Agar plate culture of Bacillus cereus

Moderate growth of beta-hemolytic shiny, large,

spreading colonies, may produce grayish to
lavender color

2. Bicarbonate agar
Bicarbonate agar culture of virulent strains of
Bacillus anthracis
Inoculation of Bacillus anthracis to a
bicarbonate agar induces capsule formation
when incubated in a candle jar or CO2 incubator

3. Phenylethyl alcohol agar (PEA)

Growth pattern on PEA differentiates Bacillus anthracis from Bacillus cereus. Bacillus
anthracis is not capable of growing on PEA when incubated at 36-370C while Bacillus
cereus can grow on the said medium