Clostridium botulinum

and the botulinum toxin:

the most potent toxin
known to mankind
Joseph An
Steven Winans
BIOMI 2600 Microbiology of Infectious Diseases
C. botulinum: An introduction
Gram-positive and anaerobic
bacteria
Endospore-producing capability
(Shukla and Sharma 2005)
Heterogeneous species (Groups
1-4) that are identified by their
collective ability to produce
botulinum neurotoxin (Peck
2009)
Cases are rare but cause serious
paralytic illness (CDC 2014) Adapted from the Textbook of
Bacteriology (top) and the CDC
C. botulinum: Structural Biology and
Environmental Niche
Rod/Bacillus-shaped
Trophic to gastrointestinal tract
Secretion of botulinum toxin into bloodstream
Inactive ovular spore form when under
suboptimal conditions
pH range = 4.6 7.0
low oxygen
Tolerates trace level of oxygen due to antioxidant
enzyme superoxide dismutase (Doyle 200)
Dormant and highly resistant spore
Temperature, UV irradiation, desiccation,
chemical damage resistance

Adapted from Cornell University Department of

 C. botulinum: Phylogeny
Group 3
Group 4

Group 1

Group 2

Adapted from the University of

Wisconsin

Adapted from Peck 2009

C. botulinum: Botulinum Toxin
Deadliest toxin known to mankind
8 antigenically distinguishable forms
(A, B, C1, C2, D, E, F, G) (Nigram &
Nigram 2010)
Light chain (active component) and
heavy chain
Adapted from Tebeda et al.
Released as a response to 2008
germination of endospore
Increased density of spores in a
given area will promote germination
and toxin release
C. botulinum: Botulinum Toxin
(cont.)
Lethal dose = 1.3-2.1 ng/kg in
humans (Ryan & Ray 2010)
Botulinum toxin is endocytosed
at the neuromuscular junction
by neuronal cells (heavy chain-
mediated)
Light chain (zinc-dependent
endopeptidase) enzymatically
cleaves SNARE proteins critical
for vesicle fusion with terminal
bouton Adapted from Tebeda et al.
Prevention of acetylcholine 2008
release (Shukla & Sharma 2008)
C. botulinum:
Economic/Health
Impact
161 laboratory-confirmed + 16 probable cases in US
(2004 alone).
Can be acquired from
Contaminated food (15) + Infant Botulism (128)
Consumption of endospores
Exposed wounds (16)
Colonization of C. botulinum on wound

Morality rate = 5-10% (treated) or 40-50% (untreated)

High economic risk
Food-borne infections may prompt a mass-produced food
product(s) to be withdrawn from markets
Infected meat product, improperly packaged food
products (canned foods)
Many vertebrates and invertebrates susceptible to
botulism
Common in poultry such as chicken, ducks, and waterfowl
(Humeau, Grant, & Poulain 2000)

Adapted from CDC

C. botulinum:
Economic/Health
Impact

Adapted from CDC

C. botulinum: Morbidity
and Host Defense
Ingestion by adults does not always lead to
illness
C. botulinum must be able to successfully
colonize the GI tract
Patients under antibiotic treatment with a
severely compromised microflora may be more
susceptible to C. botulinum infection
Increased susceptibility in infants due to lack of
microflora in infants gastrointestinal tract (Peck
et al. 2010)
Besides nonspecific innate immunity
(microflora, mucosal barrier), no specific Adapted from Bajzer and Seeley 200
host defenses against C. botulinum are
known (Wells and Wilkins)
C. botulinum: Symptoms
Acute and Severe systemic muscle fiber
paralysis (Shukla & Sharma 2008)
Difficulty breathing
Double and blurred vision
Drooping eyelids
Slurred speech
Muscle weakness (including difficulty
breathing)
Can be fatal if paralysis reaches breathing
muscles
Adapted from Fred & van Dijik
Paralysis progressively occurs top-down
Treatment?
Antitoxin (horse-derived anti-botulism toxin
immunoglobulin) that neutralizes action of
botulinum toxin circulating in the blood
(CDC 2012)
C. botulinum: Diagnosis
Spinal fluid analysis helps rule
out possibility of other CNS
disorders
Stool/wound culture test for
C. botulinum
Electromyograhy (20-50 Hz)
Mouse Toxigenicity Assay

Adapted from
http://130.237.83.53/medicald
evices/album/Ch
%206%20Measurement
%20methods%20&
%20values/index.html
C. botulinum: Prevention
Preserved food should be
heated above 248 degrees
Fahrenheit (120 degrees
Celsius) for at least 5 minutes
Discard canned goods that
have bulging lids or a foul
odor
Do not need infants
honey/corn syrup to infants Adapted from reddit.com
less than 1 year old