CLOSTRIDIUM

 Clostridia are gram positive, endospore forming, anaerobic

rods
 They are commonly found in the soil or gastrointestinal tract
of animals
 Produce ENDOSPORES
 Catalase negative, oxidase negative, non-capsulated
 They produce three major types of disease
 Histotoxic
 Enterotoxic
 Neurotoxic
Where do they live?
 Their distribution varies with the species involved but may
be found in soil, freshwater and marine sediments.
 Some appear to be common inhabitants of the intestinal
tract of man and other animals. For some species, the
intestinal canal is the main habitat and their presence in soil
is due to faecal contamination.
 They persist in ecological niches with a suitably low
oxidation-reduction potential, and can survive adverse
environments as SPORES.
How well do they survive?
 As vegetative cells they are no more resistant to heat than any
other vegetative organism.
 In the sporing stage all members have a pronounced but variable
resistance to heat. Spores of C.botulinum, for example, withstand
boiling for 3 to 4 hours. C.novyi spores are less resistant than
C.botulinum. Boiling for less than 5 minutes destroys C.perfringens
spores. It is the presence of these resistant spores that makes
autoclaving an essential part of sterilization of instruments for
surgery.
 All Clostridia are sensitive to PENICILLIN although their sensitivity
to other antimicrobial agents is NOT predictable.
How do they cause disease?
 Clostridia produce a large variety of different EXOTOXINS,
which have a wide range of properties.
 Exotoxin production may be encoded on genomic DNA, but
more frequently is associated with plasmids or phages.
 Clostridial diseases can be divided into three broad
categories:
 HISTOTOXIC –toxin in tissues (muscle, udder, liver)
 ENTEROTOXIC –toxin in blood but absorbed from intestines
 NEUROTOXIC –toxin in nerves
HISTOTOXIC CLOSTRIDIA
 GAS GANGRENE/ MYONECROSIS/ MALIGNANT EDEMA

 The name “gas gangrene” is due to the production of gas by the

Clostridia (mainly hydrogen and nitrogen), leading to the classical
“crepitus” (feels like “bubble wrap” when you touch the skin)
 The organisms then multiply, produce their toxins and enzymes.
The lesion spreads by continued growth of the bacteria in
conditions created by toxin activity ie. necrosis resulting from
reduced blood supply and reduced tissue oxidation - reduction
potential which follows oedema.
 The pathology of gas gangrene due to C.perfringens is probably
the result of alpha toxin but other proteases, eg. DNases,
collagenases, may be involved.
 PATHOGENESIS
Spores enter wound often secondary to trauma  If
conditions are right germination of spore takes place 
EXOTOXINS especially alpha, cause muscle necrosis
allowing more replication of bacteria  Factors such as
collagenase and gas cause further muscle damage 
Exotoxins and tissue products are absorbed into circulation
causing widespread damage to endothelial and blood cells,
liver and muscle  DEATH
 BLACKLEG
 The name 'blackleg' is specifically reserved for
emphysematous myonecrosis due to C.chauvoei, mainly
in cattle (but sometimes sheep) and frequently in the
absence of apparent penetrating wounds
 INFECTIOUS NECROTIC HEPATITIS (BLACK DISEASE)

 The name “Black disease” arises from the darkened

undersurface of the skin of affected animals (especially
 seen after drying of the hide), due to engorgement of
subcutaneous blood vessels.
 Lesions are seen in the liver of sheep (and on rare occasions
in pigs, cattle, horses, dogs, cats). It has been suggested that
the spores of C.novyi are lying dormant in the liver waiting
for the appropriate damage to be inflicted, which allows
them to germinate.
 The most usual trigger in sheep is the wandering of liver
fluke larvae.
 However, neoplasms and other tissue destructive lesions
may result in the multiplication of C.novyi and death
 PATHOGENESIS
Organism are ingested and spores lodge in the liver 
Migrating liver fluke larvae cause local necrosis  Lower
redox potential allows germination and multiplication 
Toxins, especially alpha, enter circulation  Increased
capillary permeability, damage to muscles including heart 
Congestion of subcutaneous vessels  Sudden Death
 Diagnosis Of Histotoxic Clostridial Disease
1. Clinical features and pathology - gas, necrosis, haemorrhage,
toxaemia and death.
2. Visualisation of organisms in tissues - Impression smears should be
made of necrotic muscle, liver, or other affected tissue. Diff Quik
(Giemsa) stains will show large, thick rods that may or may not have
spores. Gram staining will show these as being gram positive, gram
variable or gram negative. On careful inspection, a range of other
organisms may also be seen.
3. Fluorescent antibody (FAb) tests performed on impression smears for
C.chauvoei and C.septicum (which share spore antigens) used to be
available through EMAI but are no longer available.
 Treatment And Control
 If an animal is diagnosed with histotoxic clostridial
disease in time (ie. before it dies!), urgent and
aggressive debridement of the wound should be
implemented.
 It is necessary to remove all the necrotic tissue in which
the organisms are multiplying, and to re-establish
blood supply to the affected area
 Treatment has to be given early to animals and should
include PENICILLIN and ANTITOXIN. If the condition is
left even a few hours without appropriate therapy, the
animal will die before the diagnosis is confirmed.
 As many of these affected animals may simply be
found dead, the only effective management of this
type of disease is preventive VACCINATION programs
2. ENTEROTOXEMIA / ENTEROPATHY
DYSENTRY / ENTERITIS
ENTEROTOXEMIA (PULPY KIDNEY)
 This is a special category of disease that manifests as a
SYSTEMIC (TOXAEMIC) DISEASE - even though the multiplication
of the organisms occurs locally in the intestine it seems to produce
very little damage at that site.
 Disease of sheep and goats found worldwide. It is particularly a
problem with feedlot lambs fed rich rations of grains
(“overeating disease”, feedlot animals) or lambs (3-10 weeks
old) suckling heavily lactating ewes grazed on lush pastures. The
major toxin elaborated by C.perfringens Type D, which is the
most common cause of pulpy kidney in Australia, is the epsilon
toxin.
 PATHOGENESIS
Change in food conditions (rich diet, increases amount of undigested
starch in intestines and reduces peristalsis)  Rapid proliferation of
C.perfringens Type D  Epsilon prototoxin released and activated by
trypsin and chymotrypsin  Large amount of active epsilon toxin
accumulates and increases intestinal permeability  Epsilon toxin is
absorbed into circulation  Epsilon toxin increases capillary
permeability in brain causing oedema, degeneration and necrosis of
neural tissue. Also causes damage to capillaries in Loop of Henle and to
the renal tubules resulting in glycosuria  Lesions in brain stimulate
catecholamine release from adrenal medulla, causing glycogenolysis and
hyperglycaemia  Death with convulsions and hypovolaemia, oedema
of lungs, pericardium and other cavities
 This protoxin of course, is also normally produced in the large
intestine but there is not sufficient trypsin there to make it active.
In the small intestine there are specific receptors on the
enterocytes that facilitate absorption of the toxin into the
circulation.
 Once in the circulation, the toxin causes necrosis of cells and
damage to vascular endothelial cells, resulting in the local
liquefactive necrosis of brain tissue, necrosis of renal cortex
(“pulpy kidney – this is the result of rapid post-mortem autolysis
in th toxin damaged tissue), and perivascular oedema in
meninges and brain
Diagnosis Of Pulpy Kidney
 Visualisation of large numbers of the organisms in the
upper small intestine may be a useful indicator. Collect
samples quickly after death, otherwise organisms from
the large intestines will enter the small intestines and
multiply.
 Identification of the epsilon toxin in the small intestinal
contents (last 1 to 1.5 m of ileum) or serum of affected
individuals is required (counterimmunoelectrophoresis or
ELISA tests used).
 Treatment and Prevention
 There is no specific treatment, as animals are usually
found de
 Routine vaccination with a toxoid (active against the
effects of the epsilon toxin) should be carried out to
ensure that the flock is protected.
NEUROTOXIC
BOTULISM
 neurological disease
 As well as the neurotoxin, C.botulinum possesses a
haemagglutinating factor that has been associated with
vascular thrombus formation. Respiratory failure is the
usual cause of death.
 PATHOGENESIS
Toxin is absorbed from its site of entry to the body (usually the
gastrointestinal tract, but possibly also wounds)  Transported
to neurones via the blood stream  Circulating toxin reaches
susceptible nerve endings but can only bind at the
neuromuscular junctions or nerve-nerve junctions (cholinergic
junctions of peripheral nerves)  The toxin acts
presynaptically and blocks acetylcholine release  Flaccid
paralysis, respiratory depression, possible vascular thrombus
formation
The toxin that causes botulism may be acquired in three main ways:
1. FORAGE POISONING (FOOD POISONING)
 The toxin is preformed in food and is subsequently ingested. This
usually involves high protein foods (which are rich in proteases) or
foods contaminated by rotting cadavers (dead rats or cats),
vegetation or other foodstuffs.
 The rotting provides the nutrients and the enzymes, if required, to
enable full activation of the toxin before it is ingested.
 Animals can acquire this form of botulism from improperly stored
silage, silage or hay contaminated by the bodies of small rodents
which are a good source of protein and enzymes, or from carcasses
etc
2. WOUND BOTULISM
 Rare form of intoxication
TOXICOINFECTIOUS BOTULISM
 in foals less than 8 months old - usually 3 - 4 weeks.
The disease is contracted by ingestion of spores from
food such as honey
CLINICAL SIGNS
 In general it is seen as an ascending flaccid paralysis.
 Response to painful stimuli may persist. Ultimately
death is caused by respiratory paralysis and failure.
 Signs of decreased cholinergic function may also be
observed (eg. constipation, urinary retention,
decreased salivation and lacrimation, dilated pupils)
Diagnosis
 A tentative diagnosis can usually be made based on
history and clinical signs.
 Confirmation of the diagnosis of botulism is based on
finding toxin in the serum, faeces, vomitus or samples of
suspected food
Treatment and Prophylaxis
 Antitoxin (if available) – must be given early in disease and
must be of the correct Type eg most cases of C.botulinum in dogs
are type C therefore the equine product with antibodies to types
A,B and E is of no use.
 Supportive therapy - fluids, rest, nutritional support, artificial
ventilation.
 Antibiotics – while metronidazle or penicillin based antibiotics
are given in an attempt to reduce any intestinal population of
C.botulinim, this is done with the understanding that most cases of
botulism are the result of ingestion of preformed toxin.
TETANUS
 Spores of C.tetani are found worldwide in the soil
(especially heavily manured soils) and in the intestine of
man and other animals,
 PATHOGENESIS
Spores are introduced into the animal via deep penetrating wound with
minimal trauma  Germination of the spore occurs, leading to production of
small numbers of Clostridium tetani vegetative bacteria which produce LARGE
volumes of tetanus toxin at the deep wound  Tetanus toxin is transported
within the axons of the peripheral motor and sensory fibres (intra-axonal
retrograde transport) to the CENTRAL NERVOUS SYSTEM  Tetanus toxin has a
high affinity for di and tri- sialogangliosides in cells from all parts of the nervous
system but preferentially the spinal cord. Tetanus toxin acts centrally rather than
locally  The tetanus neurotoxin acts specifically on the inhibitory synapses,
preventing the release of glycine and GABA from the inhibitory interneurones in
the brain and spinal cord. The lack of inhibition leads to overexcitation of
motor neurones that manifests as increased muscle tone, rigidity and spasm
(SPASTIC PARALYSIS)
Diagnosis
 Usually made on clinical grounds alone. Isolation of
organisms is usually extremely difficult or impossible
even if the location of the wound is known (little
multiplication).
 May follow castration, shearing, docking, mulesing,
injections etc. Docking by use of elastic band ligatures
is especially hazardous.
Clinical signs
 Incubation period usually 3–10 days but could be as
long as several months
 advanced disease is characterised by convulsive
contractions of voluntary muscles
Treatment
 If a wound that may be contaminated with tetanus
spores is noticed soon after it has occurred, tetanus can
be prevented by ensuring that an anaerobic
environment does not develop. Debridement of all
necrotic tissue, exposure to oxygen and use of
chemoprophylaxis is important. In practice, the use of
penicillin (both locally within the wound, and
systemically) and injection of antitoxin should be
practiced in any animal
 Antitoxin cannot neutralise the toxin already fixed to
the nervous tissue but is able to neutralize toxin that is
still in the muscle tissue.
 Be mindful that medications may not get to intended
sites when blood supply and tissue penetration is poor
as in areas of necrosis, which is why debridement is
needed if the site of origin can be identified.
 Muscle relaxation may be achieved by use of
diazepam, acetyl promazine or one of the longer
acting barbiturates
 Supportive treatment such as i/v fluids administered if
the animal is unable to eat or drink
Prevention
 At risk animals can be vaccinated with a TOXOID
 Toxoid vaccines : A vaccine made from a toxin
(poison) that has been made harmless but that elicits an
immune response against the toxin. are based on the
toxin produced by certain bacteria (e.g. tetanus or
diphtheria