BACILLUS

DR MAHFUJA BEGAM
Assistant Professor
Department of Microbiology. Pabna
Medical college ,Pabna.

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 TLO
• Classification of gram positive bacilli
• Morphology, culture characteristics, pathogenesis of
the disease , clinical and laboratory diagnosis of:
– Bacillus
– Clostridium

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CLASSIFICATION OF GRAM POSITIVE
BACILLI

SPORE FORMING NON-SPORE FORMING

Corynebacterium
Listeria
Gardnerella
Lactobacilli
Actinomyeces

Aerobic Anaerobic
Bacillus Clostridium

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 Important properties of Bacillus
• Gram positive bacilli, spore forming and aerobic

• Uniquitous saprophytes excepts B. anthacis &B.

cereus.

• Motile with peritrichous flagella (except-B.anthracis)

• Haemolysis common (except B.anthracis)

• Spore can withstand dry heat-140ᵒc for 3 hrs,

Persist for years in dry environment.

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Gram positive spore forming Bacilli

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 Bacillus
Classification of Bacillus:
A)Pathogenic-
• Bacillus anthracis: Zoonotic disease transmitted from
infected animal
• Bacillus cereus: Food poisoning, caused by spore
contaminating rice survived in steaming
• B)Non pathogenic-
• Bacillus subtilis
• Bacillus stearothermophilus
• Bacillus.pumilus

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 Antigenic structure

• Capsular antigen
• Cell wall antigen
• Somatic antigen-Protective antigen

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 Virulence factor of B.anthracis
1. anthrax toxin: edema factor and lethal factor(A–B
subunit;B, or binding, subunit is protective antigen)
-LF+PA form lethal toxin: cause of death in infected
cells
-EF+PA forms edema toxin: responsible for cell and
tissue edema(an adenylate cyclase that causes an
increase in the intracellular concentration of cyclic
AMP. This causes an outpouring of fluid from the cell
into the extracellular space, which manifests as
edema.
2. Capsule-antiphagocytic
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 Anthrax(Bacillus anthracis)
• Cutaneous anthrax’(95%): Painless ulcer (malignant pustule)

• Pulmonary anthrax(5%): Wool sorter’s disease which can

leads to more severe complication like hemorrhagic
mediastinitis

• Gastrointestinal anthrax: Vomiting abdominal pain, bloody

diarrhea

• Lab diagnosis:
– Microscopy: Gram positive bacilli
– PCR: For rapid diagnosis

• Antibiotic: Ciprofloxacin, Doxicycline

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TRANSMISSION-B. anthracis
• entry of spores through
injured skin( spores on
animal products, such as
hides, bristles, and wool -
cutaneous anthrax)

• by inhalation of spores
into the lung (inhalation
anthrax)
• Ingestion of
contaminated meat
(Gastrointestinal anthrax )
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 Pathogenesis of Pulmonary anthrax: Wool
sorter’s
• In inhalation anthrax (woolsorters’ disease), the spores from
the dust of wool, hair, or hides are inhaled; phagocytosed in
the lungs

• and transported by the lymphatic drainage to the

mediastinal lymph nodes, where germination occurs.

• followed by toxin production and the development of

hemorrhagic mediastinitis and sepsis, which are usually
rapidly fatal.

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 Pathogenesis of Cutaneous anthrax(malignant pustule)

• Generally occurs on exposed surfaces of the arms or hands

followed in frequency by the face and neck.

• Spores germinate in the tissue at the site of entry, and growth

of the vegetative organisms

• Secretion of anthrax toxin and formation of a gelatinous

edema and congestion.

• Bacilli spread via lymphatics to the bloodstream, and they

multiply freely in the blood and tissues.

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 Pathogenesis of Cutaneous anthrax cont’
• A pruritic papule develops 1–7 days after entry of the organisms
or spores through a scratch.

• Initially, it resembles an insect bite. The papule rapidly changes

into a vesicle or small ring of vesicles that coalesce, and a
necrotic ulcer develops. The lesions typically are 1–3 cm in
diameter and have a characteristic central black eschar. Marked
edema occurs.

• Lymphangitis, lymphadenopathy, and systemic signs and

symptoms of fever, malaise, and headache may occur.

• After 7–10 days, the eschar is fully developed and dries,

loosens, and separates; healing is by granulation and leaves a
scar. 14
Malignant pustule

Wool sorter’s disease

Box-car like Gram

positive rods in chains

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• I.P-in inhalation anthrax may be as long as 6 week
• Early clinical manifestations are associated with marked
hemorrhagic necrosis and edema of the mediastinum.

• Substernal pain may be prominent, and there is pronounced

mediastinal widening visible on chest radiographs.
• Hemorrhagic pleural effusions follow involvement of the pleura;
cough is secondary to the effects on the trachea.
• Sepsis occurs, and there may be hematogenous spread to the
gastrointestinal tract, causing bowel ulceration, or to the
meninges, causing hemorrhagic meningitis.
The fatality rate in inhalation anthrax is high
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 Laboratory Diagnosis

• Specimen: fluid or pus from a local lesion, blood, pleural fluid,

and CSF in inhalational anthrax associated with sepsis and stool
or other intestinal contents in the case of gastrointestinal
anthrax

• Gram stain: Smears show large, gram-positive rods in chains

• Culture: BA-nonhaemolytic. NA: Colonies are roundgray to

white, tenacious with a rough texture and have a “cut glass”
appearance in transmitted light. Comma-shaped out growths
(Medusa head, “curled hair”) may project from the colony
• Gelatin subculture: Inverted fire tree appearance
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• Serology:
i)Direct fluorescent antibody test to detects
antigens.
ii) enzyme-linked immunosorbent assay (ELISA)
test for antibodies.

• Pronounced mediastinal widening visible on

chest radiographs.
• Polymerase chain reaction (PCR)
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 Prevention
• Ciprofloxacin or doxycycline was used as
prophylaxis in those exposed during the
outbreak
• People at high risk can be immunized with
vaccine containing purified protective antigen

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 Bacillus cereus food poisoning
• Mediated by enterotoxin

• Causes ADP ribosylation of G protein which stimulates

adenylate cyclase leading to increase concentration of cAMP
in enterocytes
• Food poisoning caused by B cereus has two distinct forms,
the emetic type, which is associated with fried rice, milk, and
pasta, and
the diarrheal type, which is associated with meat dishes and
sauces.
• Emetic form is manifested by nausea, vomiting, abdominal
cramps, and occasionally diarrhea and is self-limiting,

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• Diarrheal form has an incubation period of 1–
24 hours and is manifested by profuse
diarrhea with abdominal pain and cramps;
fever

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THANK YOU

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 Clostridium

• Gram positive, anaerobic spore forming bacteria

• Four important species

• Clostridium tetani: Tetanus
• Clostridium perfringens: Food poisoning, gas gangrane
• Clostridium botulinum: Botulism
• Clostridium difficile: Pseudomembranous colitis

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 Tetanus
• Transmission: Contaminated wound, umbilical cord

• Contaminated spore germinates in necrotic tissue with low

blood supply leading to vegetative bacteria

• Toxin (tetanospasmin) produce by the vegetative bacteria

blocks release of inhibitory neurotransmitter (glycin and
GABA)-- increase muscle spasm due to unopposed Ach

• Treatment: Tetanus immunoglobulin

• Prevention: Tetanus toxoid (Inactivated toxin)

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Tetanic spasm

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 Botulism

• Transmission: Ingestion of preformed toxins in canned

vegetables and meat

• Botulinum toxin (A.B and E) is absorbed from the gut and

transmitted through blood to peripheral nerve synapse and
block release of acetylcholine (flaccid type of paralysis)

• Wound botulism: Enter through wound

• Infant botulism: Enter through contaminated milk, honey

• Lab diagnosis: Toxin demonstration from leftover food

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 Clostridium perfringens :Gas gangrene

• Transmission: Wound contaminated by spore or abdominal

trauma in which vegetative bacteria in the colon or vagina
may become a source

• Released alpha toxin (lecithinase) causes damage on the cell

membrane and erythrocyte. Degradative enzyme produce gas
in tissue

• Microscopy: Large gram positive rods

• Culture: Colony shows double zone of hemolysis in blood agar
plate

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Gas gangrene

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 Double zone of hemolysis in BA

Egg yolk

Principle of Nagler’s reaction

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 Clostridium difficile
• Causes antibiotic associated pseudomembranous colitis

• Transmission: Normal colonic flora in 3% of population.

Transmitted by oral route

• Clostidium difficile produce exotoxin A and B that glycosylate

G protein. Toxin B also causes depolymerization of actin
filament leads to cytoskeletal damage and cell death

• Pseudomembrane(yellow-white) in the colonic mucosa

• Lab Dx: Detection of toxin in stool filtrate by ELISA

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Pseudomembrane in colon

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 REFERENCE
• Warren Levinson. Review of Medical Microbiology and
Immunology, 11th edition (2010). Appleton and Lange.

• Geo. F. Brooks, Karen C. Carroll, Janet S. Butel, Stephen A.

Morse, Timothy A. Mietzner. Medical Microbiology, 25th
edition (2010). Appleton & Lange.

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 QUIZZ 1

• Which of the following enzymes related to gas

gangrene

A. Coagulase
B. Catalase
C. Lecithinase
D. Hyaluronidase

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 QUIZZ 2

• Which of the following diseases is zoonotic

A. Tetanus
B. Diphtheria
C. Scarlet fever
D. Anthrax

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 QUIZZ 3

• A 30-year old man developed diarrhea three

days after admission to surgery unit. The
MOST possible causative agent for his illness is

A. Staphylococcus aureus
B. Vibrio cholerae
C. Escherichia coli
D. Clostridium difficile
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 QUIZZ 4

• Tetanic spasm is the result of

A. Blocking of excitatory neurotransmitter

B. Blocking of inhibitory neurotransmitter
C. An non spore forming bacterial infection
D. An aerobic bacterial infection

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Thank you
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