Clinical Bacteriology

Lecture 16

By FAWAD MAHMOOD
M.Phil. Medical Laboratory Sciences

1 05/19/2021
 THE GENUS
CLOSTRIDIUM
 Genus
Clostridium

There are four medically important species

1. Clostridium perfringens.the cause of gas gangrene

2. Clostridium botulinum, the cause of botulism.

3. Clostridium difficile, the cause of toxic enterocolitis.

4. Clostridium tetani, the cause of tetanus.

3 05/19/2021
 Characteristic of Clostridium genus
 Gram +

 Spore-forming bacteria

 Rods

 All clostridia are anaerobic

 Large, spore-forming, G+ bacilli

 Nature habitat is the soil or the intestinal tract of animals and humans

 Spores are highly resistant

 Cause disease primarily through the production of numerous exotoxins.

4 05/19/2021
 The species Clostridium
tetani

5 05/19/2021
 Characteristic of Clostridium tetani

 Grampositive, straight and slender rods with rounded ends.

 Round terminal spores are formed after 2-4 days of incubation.

 C. tetani is flagellated and motile.

 It has numerous peritrichous flagella.

 Capsules are not formed.

6 05/19/2021
 Characteristic of Clostridium tetani

 C. tetani requires strict anaerobic conditions.

 They weakly hemolyse on blood agar.

 They do not form acids from sugars.

 Aminoacids serve as main sources of energy

 The 10 types of C. tetani can be distinquished by specific flagellar (H) antigens.

 Antigenic types I and III most often cause tetanus in humans

7 05/19/2021
 Characteristic of Clostridium tetani

The spores of tetani

 The spores are heat-stable.

 The spores of some strains are resistant to boiling in water for up to 3 hours.

 They are killed by autoclaving at 121 C for 15 minutes.

8 05/19/2021
 Characteristic of Clostridium tetani
 C. tetani produces an oxygen-labile hemolysin -
tetanolysin. This toxin has only a negligible significance
for the pathogenesis of tetanus.

 The most important product of C. tetani is neurotoxic

exotoxin – tetanospasmin.

 The tetanospasmin production appears to be under control

of plasmid gene.

9 05/19/2021
 Transmission
 Spores are widespread in soil

 The portal of entry is usually a wound site (e.g., where a nail penetrates
the foot)

 The spores can also be introduced during “skin-popping,” a technique

used by drug addicts to inject drugs into the skin.

10 05/19/2021
 Pathogenesis
 Pathogenesis Tetanus toxin (tetanospasmin) is produced by bacteria at the

wound site.

 This polypeptide toxin

 This toxin spreads through tissue spaces into the lymphatic and vascular systems

 It enters the nervous system at the neuromuscular junctions and migrates into

the central nervous system

11 05/19/2021
 Pathogenesis
 where it binds to ganglioside receptors and blocks release of inhibitory

mediators (e.g., glycine and γ-aminobutyric acid [GABA]) at spinal synapses.

 stops nerve impulse to muscles

 spastic paralysis

 severe muscle contractions and spasms

 can be fatal

12 05/19/2021
 Tetanospasmin

13 05/19/2021
 Symptoms of tetanus
 Muscular spasms
 Inability to open the mouth properly

 Difficulty in mastication due to rigidity of masticatory muscles.

 High temperature.

 The patient cannot open his mouth, this effect is named as trismus.

 Facial spasm.

 In severe cases, spasms of the back muscles

 The patients are fully conscious, and pain may be very intensive.

 death

 Incubation: approximatelly 5 to 15 days

 Neonatal Tetanus

 Affects newborns without passive

immunity from their mother
 Usually due to infected umbilical
stump
 Rare in U.S., but prevalent in
underdeveloped countries

Neonatal Tetanus ID# 6374 (1995). Source: Center for Disease

Control--Public Health Image Library
 Laboratory Diagnosis
There is no microbiologic or serologic diagnosis.
Organisms are rarely isolated from the wound site.

16 05/19/2021
 Treatment
 Tetanus immune globulin (tetanus antitoxin) is used to neutralize the toxin.

 The role of antibiotics is uncertain.

 If antibiotics are used, either metronidazole or penicillin G can be given.

 An adequate airway must be maintained and respiratory support given.

17 05/19/2021
 Prevention
The best way to prevent Tetanus is by vaccination
Virtually 100% efficacy rate in properly immunized
people

18 05/19/2021
 Clostridium botulinum

19 05/19/2021
 Transmission
Spores, widespread in soil
Contaminate vegetables and meats.
germinate in the anaerobic environment.

20 05/19/2021
 Clostridium botulinum
 Grampositive bacillis

 C. botulinum is a strictly anaerobic

 It is motile with peritrichous flagella.

 Its spores are oval and subterminal.

 It is a widely distributed saprophyte occuring in soil, vegetables,

fruits, and others.

 its ability to produce a potent neurotoxin in food,

 Resistance of its spores to inactivation

21 05/19/2021
 Pathogenesis

1. Botulinum toxin can be ingested or absorbed through contact with

mucous membranes or through break in skin.

2. Botulinal toxin prevents release of neuro-transmitter acetylcholine,

resulting in inhibition of muscle movement

3. inhibits nerve impulses

4. Paralysis

5. Respiratory and cardiac failure

 Clinical Findings
weakness
 Paralysis,
 Diplopia,
 Dysphagia,
Respiratory muscle failure,
 No fever is present

23 05/19/2021
 Two special clinical forms occur
 (1) wound botulism:

 in which spores contaminate a wound, germinate, and produce toxin at the site

 (2) infant botulism, in which the organisms grow in the gut and produce the

toxin there. Ingestion of honey containing the organism is implicated in

transmission of infant botulism. Affected infants develop weakness or paralysis

and may need respiratory support

 but usually recover spontaneously.

24 05/19/2021
 Laboratory Diagnosis
 The organism is usually not cultured.

 Botulinum toxin is demonstrable in uneaten food and the patient’s

serum by mouse protection tests.

 Mice are inoculated with a sample of the clinical specimen and will

die unless protected by antitoxin.

 Enzyme-linked immunoassay (EIA)

 Polymerase-chain reaction (PCR)

25 05/19/2021
Safe Work Practices

 Personal protective equipment

Gloves

Gown

Eye protection

Facial shield

Biological safety cabinet