Bacillus anthracis

• Anthrax = primarily disease of herbivorous animals

(sheep & cattle, horses, hogs & goats)
• A gram (+), aerobic, spore forming bacillus
• First isolated by Robert Koch in 1877
• Humans accidentally encounter the disease in an
agricultural setting, usually dev’t of skin infection that
become generalized

Morphology:
• In smears from blood & tissues of infected animal=
organism usually in singly or in pairs
• Ends appear square, corners so sharp, when in chain,
leaving an oval opening between the organisms
•Encapsulated during growth in infected animal, but
capsules can’t be demonstrated in vitro unless
the organisms are cultured on a bicarbonate
containing medium in the presence of 6% CO2

•Spores are formed in culture, in soil, & in the tissues

& exudates of dead animals but not in the blood
or tissues of living animals
Cultural Characteristics:
• Inoculated on 5% blood agar plates, blood free of
antibiotics= demonstration of characteristic colonial
morphology
• pH 7.0 to 7.4 under aerobic conditions = maximal
growth
• 37°C = optimal temp for maximal growth
• After 24 hrs incubation: large, raised, opaque, grayish
white, plumose colonies, 2-3 mm in diameter, with
irregular, fringelike edge
• No hemolysis is produced
Laboratory identification:
•Virulent strains are the only organisms that produce
rough colonies when grown in the absence of
increased CO2 & mucoid colonies when grown
on sodium bicarbonate medium in an
atmosphere of 5% CO2

•Differential susceptibility of B. anthracis and B.cereus

to penicillin is the basis for the string of pearls
reaction that separates virulent and avirulent B.
anthracis from other aerobic spore formers
• After 3-6 hrs. incubation on surface of solid medium
containing 0.5ug/ml penicillin: cells become large &
spherical & occur in chains, when viewed on agar
surface resemble string of pearls
• Confirmatory Test: inoculation of a suspension of
organisms from an agar plate to the mouse =
death after 2-5 days; organisms recovered from
heart blood

Resistance:
•Due to spores, extremely resistant to adverse
chemical & physical environment
•Temp of 120°C for 15 mins. = inactivate the spores
•Spores remain viable for years in contaminated
pastures & remain a source of infection for long
periods of time
Antigenic Structure:
• 3 antigens:
a. capsular polypeptide = with high molecular wt
consisting exclusively of D-glutamic acid
b. polysaccharide somatic antigen = component of the
cell wall & contains equimolar amounts of N-
acetylglucosamine & D-galactose
c. complex protein toxin

Determinants of Pathogenicity:
• Depends on 2 impt. virulence factors:
a. poly (D-glutamic acid) capsule
b. exotoxin
• Capsules interfere with phagocytosis and impt in early
stages of infection
• Antibodies against the capsular antigen are produced,
but not protective against the disease
• Lethal effects of exotoxin consists of 3 distinct &
serologically active proteins:
a. protective antigen (PA)
b. edema factor (EF)
c. lethal factor (LF)
• Proteins act synergistically to produce systemic
effects of anthrax
• No toxic activity if functions individually
• Toxic effects appear when:
> PA + EF = produces localized edema in the skin of
test animals
>PA + LF = death in test animals
Clinical Infection:

• Soil = ultimate reservoir of anthrax infection

• Terminal stages of the disease=bacilli are shed in
large no. from all orifices of the infected animal
• In animals, anthrax is severe & usually takes the
form of septicemia
• In humans: agricultural cases = result from direct
contact with animals dying of anthrax industrial
cases = result from contact with contaminated
animal products
Pathogenesis:

Humans become infected in one of three ways:

1. Cutaneous route = organisms gain access thru
small abrasions or cuts & multiply locally with a
rather dramatic inflammatory response
2. Inhalation = organisms are inhaled, multiply in the
lung, & are swept to the draining hilar lymph
nodes, where marked hemorrhagic necrosis
may occur
3. Ingestion = rarely, organisms are ingested in
infected meat, with resultant invasion &
ulceration of the GI mucosa
Clinical Manifestations:
• Cutaneous anthrax = 95% of human cases
= begins 2-5 days after infection
as small, papules that develops
w/in a few days into small
vesicles filled w/ dark bluish
black fluid
= rupture of the vesicle reveals a
black eschar at the base w/ a
very prominent inflammatory
ring of reaction around the
eschar (malignant pustule)
= lesions found on the hands,
forearms, or head
• Pulmonary infection = known as wool-sorter’s disease
= in patients who handle raw wool, hides or horsehair
= acquire the disease by inhalation of spore
= symptoms are of respiratory infection
w/ fever, malaise, myalgia &
unproductive cough
= after several days will develop into a
severe infection with marked
respiratory distress & cyanosis---
death occurs after 24 hrs
• GI infection = nausea, vomiting & diarrhea
= occasionally hematemesis
= usually followed with shock and death

• Anthrax infection in humans provides permanent immunity;

second attacks are rare
Laboratory Diagnosis:
• Specimens for culture should be obtained from malignant
pustule, sputum or blood
• Gram stain & fluorescent-antibody stain= useful in
making presumptive diagnosis

Treatment:
• Penicillin = curative for cutaneous anthrax
• Pulmonary anthrax = diagnosis is usually made post-
mortem; same w/ GI anthrax
= if diagnosis is made in sufficient
time, give large doses of
Penicillin IV
= in patients allergic to penicillin, erythromycin or
tetracycline can
be given
Prevention:

• Active immunization = to herbivorous animals

= the only known method of
preventing anthrax

• For the protection of humans in high risk work

situations, a non-living vaccine consisting of
aluminum hydroxide-adsorbed supernatant
material from fermentor cultures of a toxigenic
but encapsulated strain of B. anthracis is used
= requires multiple doses over long periods of
time
 Other Aerobic Spore-
Forming Bacilli
Bacillus cereus:
• Food-borne illness
• Usually motile & not susceptible to penicillin
• Can cause 2 clinical syndromes:
a. short incubation period of 4 hrs
= severe nausea & vomiting & is frequently
mistaken for staphylococcal food
poisoning
= after the ingestion of fried rice
b. longer incubation period of 17 hrs
= abdominal cramping & diarrhea
= confused w/ clostridial food poisoning
Bacillus cereus
• Clindamycin = DOC
• Also susceptible to: aminoglycosides, vancomycin,
tetracycline & erythromycin

Other Bacillus Species:

Bacillus subtilis = present in the air, dust, brackish water
= common laboratory contaminant
= seen in bacteremias & eye infections in
heroin addicts
= usually respond to B-lactam antibiotics

Bacillus stearothermophilus
= spores are used to evaluate the efficacy
of autoclaving & other sterilization
procedures
Bacillus subtilis