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Clostridium
Gram-positive . obligate
anaerobes capable of
producing endospores which
protect them in harmful
environment . Individual cells
are rod shaped.
The spores are usually wider
than the rods, and are located
terminally or sub terminally.
Most clostridia are motile by
peritrichous flagella.while
others have a capsule like
Clostridium.perfringens
Characteristics
2

Clostridium consists of around 100 species that
include common free-living bacteria as well as
important pathogens There are five main
species
responsible for disease in
humans.
C. perfringens: gas gangrene; food poisoning
C. tetani: tetanus
C. botulinum: botulism
C. difficile: pseudomembranous colitis
C.Sordellii : can cause a fatal infection in exceptionally rare
cases after medical abortions
The shape an position of spores varies in different
species and is useful the identification of
Clostridia
*Central in Cl.bifermentans
*Sub terminal in Cl.perfringens
*Oval or terminal in Cl.tertium
*Spherical and terminal giving
drum stick appearance in Cl.tetani
4
 C. perfringens is a
relatively large Gram-
positive short fat
bacilli with blunt ends.
It is capsulate and
non-motile. Anaerobic.
It grows quickly on
laboratory media on
blood agar ( B –
Haemolytic )
5
 Liquid medium for
cultivation cooked
meat broth
 Thiglyclolate broth
 CMB contain
unsaturated fatty
acids which take up
oxygen
6
Clostridium. perfringens
Culture & identification
7

It distinguish between different
species of bacteria.
The lactose (milk
sugar), litmus (pH indicator),
and casein(milk protein)
contained within the medium can
all be metabolized by different
types of bacteria.

Milk is the first substrate used to
maintain bacteria, this test allows
for accurate depiction of bacterial
types. The addition of litmus,
other than explaining the pH type,
acts as an oxidation-reduction
indicator. The test itself tells
whether the bacterium can
ferment lactose, reduce litmus,
form clots, form gas,
 This test is done to detect the lecithinase
activity
 The M.O is streaked on the medium containing egg
yolk (contains lecithin)
 The plate is incubated anaerobically at 37 C for 24 h
 Colonies of Cl. perfringens are surrounded by zones
of turbidity due to lecithinase activity and the effect is
specifically inhibited if Cl. perfringens antiserum
containing antitoxin is present on the medium
 `
9
C. perfringens Nagler Reaction
NOTE: Lecithinase (α-toxin; phospholipase) hydrolyzes
phospholipids in egg-yolk agar around streak on right.
Antibody against α-toxin inhibits activity around left streak.
10
Positive Nagler Reaction
Procedure of Nagler Reaction
12
Anaerobic Jar
College Of Dentistry - Mosul University 13
 The organisms
associated with gas
gangrene attack soft
tissues by producing
toxins and aggressins,
and some strains of
the bacteria produce
enterotoxins and
cause food poisoning
14
 If there are pieces of necrotic tissue in
the wound, small pieces should be
transferred aseptically into a sterile
screw-capped bottle and examined
immediately by microscopy and culture.
Specimens of exudate should be taken
from the deeper areas of the wound
where the infection seems to be most
pronounced.
15
 Virulence factors
 toxins –
 alpha toxin – causes RBC rupture,
edema and tissue destruction
 Enterotoxin
 collagenase
 Hyaluronidase
 DNase
16
Clostridium tetani
Anaerobic bacteria of the genus species Clostridium it is gram
positive, slender bacillus and it has spherical terminal spores
giving drum stick appearance
It is non capsulated & motile with peritrichus flagella
It produces a potent biological toxin, tetanospasmin, and is the
causative agent of tetanus a disease characterized by painful
muscular spasms that can lead to respiratory failure and, in up
to 40% of cases, death.
 An infectious disease caused by
contamination of wounds from the
bacteria Clostridium tetani, or the
spores they produce that live in the soil,
and animal feces
 Infection follows when spores
 become activated and develop
 into gram-positive bacteria that multiply
 and produce a very powerful toxin
(tetanospasmin) that affects the
muscles.
 Tetanus spores are found throughout the
environment, usually in soil, dust, and animal waste.
 Tetanus is acquired through contact with the
environment; it is not transmitted from person to
person.
 The usual locations for the bacteria to enter the
body:
 Puncture wounds (such as those caused by rusty
nails, splinters, or insect bites.)
 Burns, any break in the skin, and IV drug access sites are
also potential entryways for the bacteria.

1. It inhibits the release of acetylcholine thus it
interferes with neuromuscular transmission.


2. Inhibition of postsynaptic spinal neurons by
blocking the release of an inhibiting mediator
 Gram +ve stains grow on blood agar media
aerobically
 Inoculation of culture in to 2 mice one is protected
with anti-toxin and the other is unprotected (dies with
typical tetanic spasms )

"C. diff", is a species
positive
-
Gram
of
of the genus
bacteria
that
Clostridium
causes diarrhea and
other intestinal
disease when
competing bacteria
are wiped out by
antibiotics.

Most common cause of nosocomial diarrhea.

Rate and severity of C. difficile-associated diarrhea
(CDAD) increasing.

Clostridium difficile is a bacterium that can cause
symptoms ranging from diarrhea to life-threatening
inflammation of the colon. Illness from C. difficile
most commonly affects older adults in hospitals or
in long term care facilities and typically occurs after
use of antibiotic medication

characteristic
features:

Clostridia are
anaerobic, spore-
forming rods (bacilli).

C. difficile bacteria can be found throughout the
environment — in soil, air, water, and human
and animal feces. A small number of healthy
people naturally carry the bacteria in their large
intestine. But C. difficile is most common in
hospitals and other health care facilities, where
a much higher percentage of people carry the
bacteria.
Reservoir
Infectious Agent
C.difficile
Means of
Transmission
Portal of entry
Susceptible Host
Chainof infection
Bowel and
Contaminated
environment
Contact
transmission from
contaminated
hands,
equipment or the
environment
Faecal/Oral
>65 years
History of antibiotic use
Recent received
healthcare
Underlying conditions
Abdominal surgery
Weakened immunity

Disruption of normal
colonic flora

Colonisation with C.
difficile

Production of toxin A
+/- B

Mucosal injury and
inflammation

Toxigenic strains
produce 2 major
toxins:

toxin A
(enterotoxin)

toxin B (cytotoxin)

Neutralised by C.
sordellii antitoxin

Watery diarrhea three or more times a day for two
or more days

Mild abdominal cramping and tenderness

Watery diarrhea 10 to 15 times a day

Abdominal cramping and pain, which may be
severe

Fever

Blood or pus in the stool

Nausea

Dehydration

Loss of appetite

Weight loss

Pseudomembraneous colitis

Perforation of the colon

Sepsis

Death
The Specimen
Fresh is best (test within 2 hours)
Liquid or loose, not solid
If unable to test within 2 hours, refrigerate
at 4 C for up to 3 days
Freeze at -70 C (not -20 C) if testing will
be delayed
Specimen quality will influence test results

Endoscopy
(pseudomembranous
colitis)

Culture

Cell culture cytotoxin
test

EIA toxin test

PCR toxin gene
detection
preventation

Note : alcohol does not kill C. difficile spores,
use of soap and water is more efficacious

Gram positive

Obligate anaerobic bacillus

Spores

Resistant to heat, light, drying and radiation

Specific conditions for germination

Anaerobic conditions

Warmth (10-50oC)

Mild alkalinity

Ingestion

Organism

Spores

Neurotoxin

Wound contamination

Inhalation

Person-to-person not documented
Three forms
Foodborne
Wound
Infant
All forms fatal and a medical emergency
Incubation period: 12-36 hours

The symptoms of botulism are similar to
those of Guillain-Barré syndrome, stroke,
and myasthenia gravis.

As a result, botulism is probably
substantially under-diagnosed.

Serum electrolytes, renal and liver function
tests, complete blood tests, urinalysis, and
electrocardiograms will all be normal unless
secondary complications occur.

The incubation period varies according to
the mode of transmission, rate of absorption
of the toxin, and the total amount and type
of toxin.

Foodborne botulism usually takes 24-36
hours to manifest itself.

Wound botulism often takes 3 or more days
to appear.

Inhalation botulism has occurred very rarely,
but incubation times may range from several
hours to perhaps days, again depending
upon the type and amount of toxin inhaled.

All four types of botulism result in symmetric
descending flaccid paralysis of motor and
autonomic nerves always beginning with the
cranial nerves. These symptoms are
preceded by constipation in cases of infant
botulism.

Symptoms include:

Double or blurred vision

Drooping eyelids

Dry mouth

Difficulty Swallowing

Muscle weakness

If left untreated symptoms may expand to
include paralysis of respiratory muscles as
well as the arms and legs.

Asphyxiation due to respiratory paralysis is
the most common cause of death in
botulism cases.

Proper food preparation is one of the most
effective ways to limit the risk of exposure to
botulism toxin.

Boiling food or water for ten minutes can
eliminate some strains of Clostridium botulinum
as well as neutralize the toxin as well.
However, this will not assure 100% elimination.

Limiting growth of Clostridium botulinum and
the production of botulism toxin is an
alternative to their outright destruction.

Now manufactured under the name ―Botox‖

Experimentally used for treating migraine
headaches, chronic low back pain, stroke,
cerebral palsy, and dystonias (neurologic
diseases involving abnormal muscle posture
and tension)

Frequent injections allows an individual to
develop antibodies

Studies carried out to determine feasibility of
other strains of BoNT

BoNT B manufactured for treatment of
cervical dystonia in 2000 as ―Myobloc‖
Clostridium (1).pdf

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Clostridium (1).pdf

  • 1. Clostridium Gram-positive . obligate anaerobes capable of producing endospores which protect them in harmful environment . Individual cells are rod shaped. The spores are usually wider than the rods, and are located terminally or sub terminally. Most clostridia are motile by peritrichous flagella.while others have a capsule like Clostridium.perfringens Characteristics 2
  • 2.  Clostridium consists of around 100 species that include common free-living bacteria as well as important pathogens There are five main species responsible for disease in humans. C. perfringens: gas gangrene; food poisoning C. tetani: tetanus C. botulinum: botulism C. difficile: pseudomembranous colitis C.Sordellii : can cause a fatal infection in exceptionally rare cases after medical abortions
  • 3. The shape an position of spores varies in different species and is useful the identification of Clostridia *Central in Cl.bifermentans *Sub terminal in Cl.perfringens *Oval or terminal in Cl.tertium *Spherical and terminal giving drum stick appearance in Cl.tetani 4
  • 4.  C. perfringens is a relatively large Gram- positive short fat bacilli with blunt ends. It is capsulate and non-motile. Anaerobic. It grows quickly on laboratory media on blood agar ( B – Haemolytic ) 5
  • 5.  Liquid medium for cultivation cooked meat broth  Thiglyclolate broth  CMB contain unsaturated fatty acids which take up oxygen 6
  • 7.  It distinguish between different species of bacteria. The lactose (milk sugar), litmus (pH indicator), and casein(milk protein) contained within the medium can all be metabolized by different types of bacteria.  Milk is the first substrate used to maintain bacteria, this test allows for accurate depiction of bacterial types. The addition of litmus, other than explaining the pH type, acts as an oxidation-reduction indicator. The test itself tells whether the bacterium can ferment lactose, reduce litmus, form clots, form gas,
  • 8.  This test is done to detect the lecithinase activity  The M.O is streaked on the medium containing egg yolk (contains lecithin)  The plate is incubated anaerobically at 37 C for 24 h  Colonies of Cl. perfringens are surrounded by zones of turbidity due to lecithinase activity and the effect is specifically inhibited if Cl. perfringens antiserum containing antitoxin is present on the medium  ` 9
  • 9. C. perfringens Nagler Reaction NOTE: Lecithinase (α-toxin; phospholipase) hydrolyzes phospholipids in egg-yolk agar around streak on right. Antibody against α-toxin inhibits activity around left streak. 10
  • 12. College Of Dentistry - Mosul University 13
  • 13.  The organisms associated with gas gangrene attack soft tissues by producing toxins and aggressins, and some strains of the bacteria produce enterotoxins and cause food poisoning 14
  • 14.  If there are pieces of necrotic tissue in the wound, small pieces should be transferred aseptically into a sterile screw-capped bottle and examined immediately by microscopy and culture. Specimens of exudate should be taken from the deeper areas of the wound where the infection seems to be most pronounced. 15
  • 15.  Virulence factors  toxins –  alpha toxin – causes RBC rupture, edema and tissue destruction  Enterotoxin  collagenase  Hyaluronidase  DNase 16
  • 16. Clostridium tetani Anaerobic bacteria of the genus species Clostridium it is gram positive, slender bacillus and it has spherical terminal spores giving drum stick appearance It is non capsulated & motile with peritrichus flagella It produces a potent biological toxin, tetanospasmin, and is the causative agent of tetanus a disease characterized by painful muscular spasms that can lead to respiratory failure and, in up to 40% of cases, death.
  • 17.  An infectious disease caused by contamination of wounds from the bacteria Clostridium tetani, or the spores they produce that live in the soil, and animal feces  Infection follows when spores  become activated and develop  into gram-positive bacteria that multiply  and produce a very powerful toxin (tetanospasmin) that affects the muscles.
  • 18.  Tetanus spores are found throughout the environment, usually in soil, dust, and animal waste.  Tetanus is acquired through contact with the environment; it is not transmitted from person to person.
  • 19.  The usual locations for the bacteria to enter the body:  Puncture wounds (such as those caused by rusty nails, splinters, or insect bites.)  Burns, any break in the skin, and IV drug access sites are also potential entryways for the bacteria.
  • 20.  1. It inhibits the release of acetylcholine thus it interferes with neuromuscular transmission.   2. Inhibition of postsynaptic spinal neurons by blocking the release of an inhibiting mediator
  • 21.  Gram +ve stains grow on blood agar media aerobically  Inoculation of culture in to 2 mice one is protected with anti-toxin and the other is unprotected (dies with typical tetanic spasms )
  • 22.  "C. diff", is a species positive - Gram of of the genus bacteria that Clostridium causes diarrhea and other intestinal disease when competing bacteria are wiped out by antibiotics.
  • 23.  Most common cause of nosocomial diarrhea.  Rate and severity of C. difficile-associated diarrhea (CDAD) increasing.  Clostridium difficile is a bacterium that can cause symptoms ranging from diarrhea to life-threatening inflammation of the colon. Illness from C. difficile most commonly affects older adults in hospitals or in long term care facilities and typically occurs after use of antibiotic medication
  • 25.  C. difficile bacteria can be found throughout the environment — in soil, air, water, and human and animal feces. A small number of healthy people naturally carry the bacteria in their large intestine. But C. difficile is most common in hospitals and other health care facilities, where a much higher percentage of people carry the bacteria.
  • 26. Reservoir Infectious Agent C.difficile Means of Transmission Portal of entry Susceptible Host Chainof infection Bowel and Contaminated environment Contact transmission from contaminated hands, equipment or the environment Faecal/Oral >65 years History of antibiotic use Recent received healthcare Underlying conditions Abdominal surgery Weakened immunity
  • 27.  Disruption of normal colonic flora  Colonisation with C. difficile  Production of toxin A +/- B  Mucosal injury and inflammation
  • 28.  Toxigenic strains produce 2 major toxins:  toxin A (enterotoxin)  toxin B (cytotoxin)  Neutralised by C. sordellii antitoxin
  • 29.  Watery diarrhea three or more times a day for two or more days  Mild abdominal cramping and tenderness  Watery diarrhea 10 to 15 times a day  Abdominal cramping and pain, which may be severe  Fever  Blood or pus in the stool  Nausea  Dehydration  Loss of appetite  Weight loss
  • 30.  Pseudomembraneous colitis  Perforation of the colon  Sepsis  Death
  • 31. The Specimen Fresh is best (test within 2 hours) Liquid or loose, not solid If unable to test within 2 hours, refrigerate at 4 C for up to 3 days Freeze at -70 C (not -20 C) if testing will be delayed Specimen quality will influence test results
  • 34.
  • 35.
  • 36.
  • 37.  Note : alcohol does not kill C. difficile spores, use of soap and water is more efficacious
  • 38.
  • 39.  Gram positive  Obligate anaerobic bacillus  Spores  Resistant to heat, light, drying and radiation  Specific conditions for germination  Anaerobic conditions  Warmth (10-50oC)  Mild alkalinity
  • 41. Three forms Foodborne Wound Infant All forms fatal and a medical emergency Incubation period: 12-36 hours
  • 42.  The symptoms of botulism are similar to those of Guillain-Barré syndrome, stroke, and myasthenia gravis.  As a result, botulism is probably substantially under-diagnosed.  Serum electrolytes, renal and liver function tests, complete blood tests, urinalysis, and electrocardiograms will all be normal unless secondary complications occur.
  • 43.  The incubation period varies according to the mode of transmission, rate of absorption of the toxin, and the total amount and type of toxin.  Foodborne botulism usually takes 24-36 hours to manifest itself.  Wound botulism often takes 3 or more days to appear.  Inhalation botulism has occurred very rarely, but incubation times may range from several hours to perhaps days, again depending upon the type and amount of toxin inhaled.
  • 44.  All four types of botulism result in symmetric descending flaccid paralysis of motor and autonomic nerves always beginning with the cranial nerves. These symptoms are preceded by constipation in cases of infant botulism.  Symptoms include:  Double or blurred vision  Drooping eyelids  Dry mouth  Difficulty Swallowing  Muscle weakness
  • 45.  If left untreated symptoms may expand to include paralysis of respiratory muscles as well as the arms and legs.  Asphyxiation due to respiratory paralysis is the most common cause of death in botulism cases.
  • 46.  Proper food preparation is one of the most effective ways to limit the risk of exposure to botulism toxin.  Boiling food or water for ten minutes can eliminate some strains of Clostridium botulinum as well as neutralize the toxin as well. However, this will not assure 100% elimination.  Limiting growth of Clostridium botulinum and the production of botulism toxin is an alternative to their outright destruction.
  • 47.  Now manufactured under the name ―Botox‖  Experimentally used for treating migraine headaches, chronic low back pain, stroke, cerebral palsy, and dystonias (neurologic diseases involving abnormal muscle posture and tension)  Frequent injections allows an individual to develop antibodies  Studies carried out to determine feasibility of other strains of BoNT  BoNT B manufactured for treatment of cervical dystonia in 2000 as ―Myobloc‖