Clostridial diseases

Anthrax

Clostridial diseases

Clostridium spp.

Gram positive rods

Obligate anaerobes
Endospore forming

Clostridium spp.

Endospores

Resistant to:

Heat
Disinfectants

Clostridium spp.
Soil (found on just about all
environmental surfaces)
Fecal matter of humans and
animals
Aquatic sediments

Clostridium spp. (obligate

anaerobes)

Rapidly killed by highly reactive

oxygen radicals

Singlet oxygen
Superoxide free radicals

The Clostridia genus

The clostridia produce a wide variety of
extracellular enzymes to degrade large
biological molecules in the environment
into fermentable components
Hence, the clostridia play an important
role in nature in biodegradation and the
carbon cycle
In anaerobic clostridial infections, these
enzymes play a role in invasion and
pathology

Grown in the laboratory

under anaerobic conditions

Who are the Clostridias?

Most of the clostridia are saprophytes

but a few are pathogenic for humans
Those that are pathogens have
primarily a saprophytic existence in
nature and, in a sense, are opportunistic
pathogens
Clostridium tetani and Clostridium
botulinum produce the most potent
biological toxins known to affect humans

Clostridium perfringens
C. perfringens produces a huge array of invasins
and exotoxins, causes wound and surgical
infections that lead to gas gangrene, in
addition to severe uterine infections.
Clostridial hemolysins and extracellular enzymes
such as proteases, lipases, collagenase and
hyaluronidase, contribute to the invasive process
C. perfringens also produces an enterotoxin and
is an important cause of food poisoning.
Usually C. perfringens and C. botulinum is
encountered in improperly sterilized (canned)
foods in which endospores have germinated

Clostridium difficile
Pseudomembranous colitis in humans
is caused by overgrowth ofClostridium
difficile in the colon, usually after the
normal flora has been disturbed by
antimicrobial chemotherapy.
C. difficile produces two toxins: Toxin A is
referred to as an enterotoxin because it
causes fluid accumulation in the bowel
Toxin B is an extremely lethal (cytopathic)
toxin.

Clostridium tetani

Clostridium tetani is the causative agent of

tetanus.
The organism is found in soil, especially heavilymanured soils, and in the intestinal tracts and feces
of various animals.
Carrier rates in humans vary from 0 to 25%, and the
organism is thought to be a transient member of the
flora whose presence depends upon ingestion.
The organism produces terminal spores within a
swollen sporangium giving it a distinctive drumstick
appearance.
Although the bacterium has a typical Gram-positive
cell wall, it may stain Gram-negative or Gramvariable, especially in older cells.

Neutralization of oxygen radicals

Superoxide Dismutase
(SOD)
Catalase
Clostridium spp. lacks these
enzymes

Clostridium spp.
C. tetani
C. botulinum
C. perfringens
C. difficile

Clostridium tetani
Drumstick appearance

Clostridial diseases

Tetanus
Botulism
Gas gangrene
Foodborne gastroenteritis
Pseudomembrane colitis

TETANUS
Localized infection, caused
by a C.tetani toxin
General muscle spasmsdue to neurotoxin

TETANUS (cont.)
Trauma (often trivial)
Chronic skin ulcers
Umbilical cord (clay matter
ritual in Africa)
Abortion (unsanitary
abortions)

TETANUS (cont.)

300,000/year world wide

100 cases/year in the USA (24
fatal)
70% unvaccinated, or
incomplete vaccination regime
Mostly 60 years and older

TETANUS (cont.)

Spores in wound
Infection and/or tissue
necrosis
Low O2 tension
Spore germination

TETANOSPASMIN

Blocks the muscle

relaxation pathway
Death results from spasms
of respiratory muscles
Released from dead
bacterial cells

TETANOSPASMIN (cont.)

Once it attaches to
nerves, therapy is usually
ineffective

Advanced case of tetanus

Greek tetanos, to stretch

TREATMENT OF TETANUS

Removal of necrotized
(death) tissue
Antibiotics
Human
immunoglobulins

TETANUS VACCINE

Toxin

Toxoid
Enzyme

-S-SActive

Inactive

TETANUS VACCINATION SCHEDULE

(CDC)

DTaP vaccine (Diptheria,

Tetanus & acellular
pertussis)
2, 4, 6 and 12-18 months
4-6 years

FOODBORNE BOTULISM

A non-infectious food
poisoning
C. botulinum neurotoxin
in contaminated food
34 cases in 1994 (USA)

FOODBORNE BOTULISM (cont.)

BOTULIN TOXIN

Most potent of all natural

toxins
Approx. 0.001 mg human
lethal dose
Causes flaccid paralysis
Death follows cardiac failure

Definitive Diagnosis
only establish with toxin identification:
Isolation of toxigenic cultures and
identification of the involved type C or
D toxin with the aid of serum
neutralization in mice or guinea-pigs.
Toxin detection in clinical samples
collected for laboratory analysis
(intestinal contents)

Type of tests to ID botulism

tests may include a brain scan, spinal fluid
examination, nerve conduction test
(electromyography, or EMG),
The most direct way to confirm the diagnosis is
to demonstrate the botulinum toxin in the
patient's serum or stool by injecting serum or
stool into mice and looking for signs of botulism
The bacteria can also be isolated from the stool
of persons with foodborne and infant botulism
These tests can be performed at some state
health department laboratories and at CDC

BOTULIN TOXIN (cont.)

Home preserves (pH 5 and above)

frequent source of botulism
Not formed in pH below 4.7
Molds may shift pH to above 4.7
Destroyed by boiling 10 min
Treatment of Botulism NONE

BOTULISM PREVENTION
Preservation of foods at pH below
4.7
Salt (brine) and sugar
Nitrites in cured foods (remember
the nitrites/nitrates in hot dogs,
cured ham, processed meats)
Boiling food 10 min

INFANT BOTULISM

Predominant form in the USA

75-100 cases/year in the USA

INFANT BOTULISM (cont.)

Less than 6 months old children
Associated with ingestion of honey
Honey has endospores in it
naturally bees pick up the
endospores from the flowers
Immature intestinal microflora of
infants leads to infant botulism

children may receive medical attention because

of symptoms such as constipation, poor sucking
action, a weak cry, and a general, progressive
muscle weakness.

Infant botulism detection tests!!

The diagnosis is confirmed by the
detection of the organism or its toxin
in the infants stool
Toxin isolation and identification are
accomplished via mouse lethality
testing, with typing (type C or D toxin)
confirmed by neutralization of toxin
by specific sera (antibodies
immunoglobulins)

Where are these endospores

commonly found?

More than 90% of reported cases

(infant botulism) in the USA come
from California, Utah, and southeast
Pennsylvania; this is likely a
consequence of high concentrations
of C. botulinum spores in the soil of
these regions

WOUND BOTULISM

C. botulinum develops in
grossly contaminated wounds
19 cases in 1995
Very common with black tar
heroine users/skin popping

Therapeutic use of botulinum toxin

blepharospasm and strabismus

BOTOX

What is Blepharospasm?
Blepharo means "eyelid". Spasm
means "uncontrolled muscle
contraction".
The term blepharospasm can be
applied to any abnormal blinking or
eyelid tic or twitch resulting from any
cause, ranging from dry eyes to
Tourette's syndrome to tardive
dyskinesia.

What is Strabismus?

Strabismus, also known as crossed or

turned eye, is the medical term used
when the two eyes are not straight. It
occurs in approximately 2% to 4% of
the population.

GAS GANGRENE

GAS GANGRENE

Tissue necrosis (death) from lack

of blood supply
Caused by C. perfringens
Neglected wounds (anaerobic
conditions) provide a suitable
environment for C. perfringens
growth

Gas gangrene

Clostridium perfringens

GAS GANGRENE (cont.)

Highly lethal if untreated

C. perfringens ferments muscle proteins
and carbohydrates producing H2 and CO2
Predisposing factors:

Dirt in wound
Long delay before wound care
Induced abortion

GAS GANGRENE (cont.)

Predisposing factors:

Arteriosclerosis
Diabetes

Gas
gangrene
in arm

Bone fracture with

gas in surrounding
muscular tissue

Gas gangrene
in foot

Gas
gangrene
in foot

Gas gangrene in buttocks

TREATMENT

Removal of necrotized (dead) tissue

Amputation

Hyperbaric O2
Antibiotics

Hyperbaric chamber for

gas gangrene treatment

Anthrax

Anthrax
Caused by Bacillus anthracis (aerobic;G+;
endospore former)
commonly found in the soil (South and Central
America, Southern and Eastern Europe, Asia,
Africa, Caribbean and the Middle East)
primary disease of domesticated and wild
animals-particularly herbivores
humans become infected when they come into
contact with diseased animals (flesh, bones,
hides, hair and excrement)

Robert Koch-1876, 1877

isolated and obtained a pure
culture
Louis Pasteur-1881, developed a
vaccine

Robert Koch
Koch was a doctor and he had a detailed
knowledge of the human body something
that Pasteur, as a research scientist lacked.
He was also skilled in experiments, the result
of his work in natural sciences
Qualities that also proved to be important
were his ability to work for long periods of
time and his patience
However, Koch was also difficult to work with
and could not tolerate anyone telling him
that his theories were wrong

Robert Koch

In 1872, Koch became district medical

officer for a rural area near Berlin. He
started to experiment with microbes
in a small laboratory he had built for
himself in his surgery.

Anthrax vaccine--Pasteur

In France at that time many cattle suffered

from anthrax, a serious disease from which
many of them died.
after many experiments Pasteur succeeded
in producing a weakened & harmless culture
of anthrax bacteria
He inoculated cattle & sheep with this giving
them a mild form from which they recovered
When these animals were introduced with
others who had a severe form they remained
unaffected. They were immune.

Gram Stain-Gram
Positive

Robert
Kochs
Photos

Human cases of Anthrax are rare:

1/100,000 risk

Three forms:
Cutaneous-Most common form
acquired through a cut or abrasion of the
skin, which comes into contact with spores
from the soil or a contaminated animal
Inhalation acquired by the inhalation of sporecontaining dust where animal hair or hides
are handled
Intestinal- (Speculated)
consumption of contaminated meat

Cutaneous-Spores germinate, vegetative cells

multiply and a lesion (black=necrotic tissue)
develops at the site of infection
Extreme cases involve bacteria in the
bloodstream which can be fatal (25%)
Inhalation-Symptoms may resemble a common
cold, progressing to abrupt fever and chest pain.
After several days, severe breathing problems
and shock, resulting in death (hemorrhage)
leads to 100% death if left untreated
Intestinal-Inflammation of the intestinal tract,
nausea, loss of appetite, vomiting, severe
diarrhea and death (25-60%)

Cutaneous Anthrax

Treatment:
Antibiotics
Effective if given within 24 hours, or before
the bacteria enter the bloodstream
Penicillin, Tetracyclines, fluoroquinolones
(Cipro)
Vaccine
protective antigen (composed of a fraction
of the toxin)
95% protective
*No evidence of person-to-person transmission

Anthrax Toxin-Symptoms and disease is caused

by a toxin
made up of a protease (protein-digesting
enzyme)
B. anthracis form endospores
Spores may survive in the soil, water and on
surfaces for many years
Destroyed by autoclaving, burning, or
chlorination

Biological Warfare

Any disease-causing organism that is used as a

weapon
Anthrax has particularly useful features to be
used as a weapon
Stable in the environment (endospores)
spores that can be inhaled
once spores are inhaled, vegetative cells
grow and produce lethal toxins