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Bacillus
Bacillus anthracis
Bacillus cereus
Bacillus subtilis
stearothermophilus
> Anthrax = primarily disease of herbivorous animals (sheep & cattle, horses,
hogs & goats)
> A gram (+), aerobic, spore forming bacillus
> First isolated by Robert Koch in 1877
brackish water
contaminant
> seen in bacteremias & eye
antibiotics
sterilization procedures
> In smears from blood & tissues of infected animal = organism usually 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 cant 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
> 37C = 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 hr 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 120C 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:
a.)
anthrax
b.) 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
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:
A. 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
bluishblack 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
B. Pulmonary infection = known as wool-sorters 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
C. 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
Treatment:
> Clindamycin = DOC
> Also susceptible to: Aminoglycosides, Vancomycin, Tetracycline
can be given
Prevention:
> Active immunization = to herbivorous animals preventing anthrax
> For the protection of humans in high risk work situations, a non-living vaccine
consisting of aluminum hydroxide-adsorbed supernatant material from
& Erythromycin
Clostridium
> Gram (+) spore-forming bacilli
> Most species are obligate anaerobes
> Pathogenic species produce soluble toxins that are extremely potent
> Present in soil & GIT of humans & animals
> The pathogenic clostridia can be divided into 4 major groups, according to the types of disease they
produce:
Enterotoxigenic
Histotoxic clostridia
clostridia
Clostridium tetani
Clostridium botulinum
> Clostridium botulinum, is
> The histotoxic clostridia
> Clostridium tetani, the
the etiologic agent of
characteristically cause a
causative agent of
botulism, w/c results from the
variety of tissue infection,
> The enterotoxigenic
tetanus, causes disease
ingestion of a powerful
usually subsequent to
clostridia produce food
thru a potent exotoxin
exotoxin previously formed by
wounds or other types of
poisoning & more severe
that is produced during
the organism in contaminated
traumatic injury
forms of GI disease
limited growth w/in tissue food
perfringens)
associated Colitis
Clostridium tetani
Clostridium botulinum
> Produces the most potent exotoxin
> Neurotoxin = cause of botulism, a severe
neuroparalytic disease characterized by
sudden onset & swiftness of course,
terminating in profound paralysis & pulmonary
arrest
> 8 serologically distinct botulinum toxins: A,
B, C1 , C2, D, E, F & G
contaminated food
sausage
1. C. perfringens
2. C. novyi
3. C. Septicum
lockjaw)
Clostridium perfringens
> Cultured from 60-90% of cases of clostridial myonecrosis
> 5 types: A to E = separated according to their production of 4
major lethal toxins
> Type A = primarily responsible for disease in humans:
clostridial myonecrosis), less severe wound infections
&
a common form of food poisoning
= found in almost all GIT of animals
Morphology:
> Young cultures = gram(+);
Morphology:
Morphology:
w/rounded end
drumstick appearance
round structure
wounds
to the organism
Cultural Characteristics:
> Cultures on BAP after 24hrs incubation: circular & smooth, 2-4
occurs on BAP
mm in diameter
w/ a delicate filamentous
advancing edge
Cultural Characteristics:
swarming
observed
hemolytic
carbohydrates
+D10
Resistance:
Resistance:
mins.
and D.
contaminated materials
Antigenic Structure:
Antigenic Structure:
Antigenic Structures:
spore antigens
D, E, F & G
> Has 12 different toxins, all are protein in nature & antigenic
antigens
Determinants of Pathogenicity:
clostridial
myonecrosis
> Toxin is a lecithinase C (or phospholipase C) w/c splits lecithin
= neurotoxin (tetanospasmin)
multiplication of
(opisthotonus)
but toxin
reaching the colon is slowly absorbed--lymphatic
system----bloodstream---functional
disturbance of the peripheral nervous
system---inhibition of the release of
acetylcholine----toxin acts on the myoneural
junction to produce complete paralysis of the
cholinergic nerve fibers at the point of release
of the acetylcholine
Clinical Infection:
> Trauma associated with deep & lacerated or crush wounds of
muscle & w/ vascular damage of major vessels & capillary beds
is one of the important factor predisposing to clostridial
myonecrosis
> The basis for the requirement of trauma w/ ischemic or
necrotic areas is the anaerobic nature of the organism, w/c
require a reduced oxygen tension & oxidation-reduction
potential for growth
Pathogenesis:
> In areas of reduced O2 tension the pyruvate of muscle is
incompletely oxidized & lactic acid accumulates, causing a drop
in pH, activates endogenous proteolytic enzymes resulting in
Pathogenesis:
tissue necrosis
> Multiplication of the organisms is accompanied by the
oxidation-
organism
organism
Clinical Manifestations:
> 3 categories of increasing levels of severity of wound
infection:
1. Simple wound contamination
2. Anaerobic cellulitis
3. Clostridial myonecrosis
_______________
1. Simple Wound Contamination:
> One or more histotoxic clostridia may be present w/o
pathologic process
> Clostridia present may be non-toxigenic
2. Anaerobic Cellulitis:
> More serious form of wound infection
Clinical Manifestations:
Clinical Manifestations:
intact muscle
lockjaw
prognosis
3. Clostridial Myonecrosis:
after.
lower extremities
early complaints
fractures
diarrhea
Uterine Infection:
w/:
spasm to tetanospasms,
> The disease progresses rapidly & has a high mortality rate
Clostridial Septicemia:
of acute abdomen
Clinical Manifestations:
convulsions
stiffness
death
Immunity:
> Adequate immunization of
pregnant patients
= important for passive immunity
for the newborn
Laboratory Identification:
> Clinical material should be
transported containing CO2.
> Inoculated immediately on both
solid media & anaerobic culture
media such as chopped meat &
incubated under anaerobic
condition
> Rapid motility of the organism is
also helpful in the isolation
> Final proof of the isolation of a
toxin production when injected into
mice & its neutralization in mice
previously inoculated with antitoxin
Laboratory Diagnosis:
> Early diagnosis must be made on clinical grounds
Laboratory diagnosis:
Laboratory Diagnosis:
> Direct smear & gram stain of material deep w/in the wound =
Laboratory Diagnosis:
procedures
extensive
the organism
specimen.
Treatment:
1. Simple wound
= removal of necrotic tissue & by cleansing
= admin. of antibiotics rarely required
2. Anaerobic cellulitis
= opening the involved area, removing all necrotic
Treatment:
antibiotics
3. Clostridial myonecrosis
be given
tetanospasm
apparatus
Treatment:
inactivated
respiratory complications
renal systems
spread of infection
visual stimuli
acetylcholine
release
Prevention:
> Early & adequate wound debridement
> Adequate cleansing, removal of necrotic tissue
> Administration of prophylactic antibiotic (Penicillin)
= reduces the risk of anaerobic infection
> Food Poisoning:
Prevention:
Prevention:
complete
(DPT)
spores
perfringens