BACILLUS

Sporogenous, rod shaped bacteria are classified

into 2 genera- aerobic Bacillus and anaerobic
Clostridia.
Genus Bacillus- aerobic bacilli forming heat
resistant spores. Gram positive, Motile with
peritrichous flagella( exception B.anthracis).
Spores are ubiquitous- soil, water, dust and air.
Commonest contaminant of culture media.
Major pathogenic sps - B.anthracis – causative
agent of anthrax.
B.cereus- causes- foodborne gastroenteritis.
 Bacillus anthracis
Morphology:
One of the largest pathogenic bacterium.
In tissues- found singly, in pairs or in short chains-
entire chain being surrounded by a capsule.
Capsule is polypeptide in nature – composed of
polymer of d- glutamic acid.
Capsules are not formed under ordinary
conditions of culture- but only if media contain
added bicarbonate or are incubated under 10-
25%CO2. or if grown in media containing serum,
albumin, charcoal, starch.
In cultures bacilli are arranged to end in long
chains. The ends of the bacilli are truncated or
often concave and swollen- ‘Bamboo stick’
appearance.

Spores are formed in culture, dead animal's

tissue or in soil but never in the animals body
during life.
Sporulation occurs under unfavourable
conditions and is encouraged by distilled
water, 2 % NaCl or growth in oxalated agar.
Oxygen is required for sporulation not for
germination.
Sporulation is inhibited by calcium chloride.
Gram positive & nonacid fast, Nonmotile .
When blood films containing anthrax bacilli are
stained with Polychrome methylene blue for few
seconds – examine under microscope-
Amorphous purplish material noticed around the
bacilli- this is the capsular material- this is
characteristic of anthrax bacilli- this is called –
M’Fadyean’s rection- Presumptive diagnosis of
anthrax in animals.
Cultural Characteristics:
Aerobe and facultative anerobe.
Opt Temp- 35-37⁰C. Opt temp for sporulation- 25-
30⁰C.
Good growth occurs on ordinary media.
On solid media- irregularly round, raised, dull ,
opaque, greyish white colonies – with frosted glass
appearance.
Under Low power microscope- edge og colony-
composed of long interlacing chains of bacilli-
resembling locks of matted hair- Medusa head
appearance.
Virulent capsulated strains form rough colonies;
Avirulent strains form smooth colonies.
On gelatin stab culture – characteristic ‘inverted fir tree
appearance’ is seen-due to slow liquefaction
commencing from the top.
On BA- Colonies are nonhemolytic : some strains
produce a narrow zone of hemolysis.
In broth – growth occurs as floccular deposit with little
or no turbidity.
When B.anthracis grown on solid media containing 0.05-
0.50 units of pencillin/ml – in 3-6 hrs the cells become –
large , spherical & occur in chains on the surface of agar-
resembling string of pearls – called as ‘string of pearls
reaction’- differentiate anthrax bacilli from B.cereus.
Selective media – PLET medium- consisting of
polymyxin, lysozyme, ethylene diamine tetra
acetic acid(EDTA) and thallous acetate – added to
heart infusion agar.
Biochemical Reactions:
Glucose, maltose and sucrose fermented with
acid but no gas.
Nitrates reduced to nitrites. Catalase positive.
Pathogenicity:
2 virulence factors identified- Capsular
polypeptide and anthrax toxin- each one coded by
separate plasmid.
ANTHRAX:
Anthrax is a zoonosis. Animals are infected by
ingestion of spores present in soil.
Infected animal shed – large number of bacilli
from mouth, nose & rectum – sporulate in soil-
and remain as source of infection.
Disease is of 3 types:
Cutaneous anthrax
Pulmonary anthrax
Intestinal anthrax
All types leading to fatal septicemia or meningitis.
Cutaneous anthrax
Entry of infection through skin.
Face , neck, hands, arms and back are the usual
sites.
Lesion starts as a papule 1-3 days after infection-
and becomes vesicular –containing fluid which may
be clear or blood stained.
Whole area is congested and edematous and
several satellite lesions filled with serum or
yellow fluid are arranged round a central necrotic
lesion – which is covered by a black eschar.
The lesion is called a Malignant Pustule.
Name anthrax – means coal – comes from black
colour of eschar.
The disease is common in dock workers carrying
loads of hides and skins on their bare backs and
hence known as – Hide porter’s Disease.
Cutaneous anthrax resolves spontaneously but in
some untreated cases – patients may develop fatal
septicemia or meningitis.
Pulmonary anthrax
Called the wool sorter’s disease – bcoz common in
dock workers in wool factories – due to inhalation of
dust from infected wool.
Hemorrhagic pneumonia with high fatality rate.
Hemorrhagic meningitis may occur as complication.
Intestinal anthrax
Rare and occurs mainly in primitive communities- eating
carcasses of animals dying with anthrax.
Violent enteritis with bloody diarrrhea. High fatality
rate.
Human anthrax- industrial or non industrial( agricultural)
 Industrial – found in workers in industries such as meat
packing or wool industries.
Non industrial – is occupational disease- veterinarians,
butchers and farmers.
Cutaneous anthrax – caused by shaving brushes made
with animal hair.
Laboratory Diagnosis
Anthrax diagnosed by – microscopy, culture, animal
inoculation and serological demonstration of anthrax
antigen in infected tissues.
Acute & convalescent phase sera – antibodies
demonstration by – gel diffusion, complement
fixation, antigen coated tanned red cell agglutination
& ELISA.
Animal suspected to die of anthrax – autopsy not
permissible – split of blood leads to contamination of
soil.
An ear lobe cut off from the carcasses – sent to lab
Swabs soaked in blood or blood smears may be sent.
Presumptive diagnosis – Gram staining &
M’Fadyeans reaction.
Immunofluorescent microscopy – confirm
identification.
Animal inoculation - bacillus isolated from
contaminated tissues by applying them over the
shaven skin of guinea pig – able to penetrate
through minute abrasions and produce fatal
infections.
Culture – on NA, BA, PLET – colony identified by
morphological, biochemical tests.
Further confirmation – PCR for anthrax bacillus
with specific chromosomal markers can be done.
Prophylaxis
Improvement of factory hygiene and proper
sterilization of animal products likes hides and
wool.
Carcasses of animals suspected to have died of
anthrax – buried deep in quicklime or cremated to
prevent soil contamination.
Treatment
Antibiotics have no effect on the toxin once it is
formed.
Doxycycline, ciprofloxacin are used.