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Clostridium

This document discusses the bacteria Clostridium. Key points include: - Clostridium is a gram-positive, spore-forming, obligate anaerobe that can cause diseases like gas gangrene (C. perfringens) and tetanus (C. tetani). - C. perfringens produces alpha and theta toxins which cause cell and tissue damage leading to gas gangrene. C. tetani produces a neurotoxin causing painful muscle spasms. - Identification involves gram stain of samples from infected tissues showing characteristic morphology and culture techniques to confirm species. Proper treatment involves antibiotics, surgery, and antitoxins.

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118 views44 pages

Clostridium

This document discusses the bacteria Clostridium. Key points include: - Clostridium is a gram-positive, spore-forming, obligate anaerobe that can cause diseases like gas gangrene (C. perfringens) and tetanus (C. tetani). - C. perfringens produces alpha and theta toxins which cause cell and tissue damage leading to gas gangrene. C. tetani produces a neurotoxin causing painful muscle spasms. - Identification involves gram stain of samples from infected tissues showing characteristic morphology and culture techniques to confirm species. Proper treatment involves antibiotics, surgery, and antitoxins.

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majoragarwal1195
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CLOSTRIDIUM

MEDICALLY IMPORTANT
BACTERIA: SO FAR..
 Gm + ve cocci  Gm negative bacilli

Staphylococci Enterobacteriaceae
Streptococci Pseudomonas
Pneumococci
 Gm positive bacilli
 Gm Neg Cocci Clostridium (obligate anaerobe)
Neisseria Corynebacterium
Bacillus (obligate aerobe)
CLOSTRIDIUM
 Obligate anaerobe
 Gram positive bacilli
 Spore forming- bulging
 Name derived from kloster- Spindle shaped
CLOSTRIDIUM
 Mostly sparophytes
 Can be normally found in human/animal intestines
 Diseases
 Gas gangrene -Clostridium perfringens

 Tetanus- Clostridium tetani

 Food poisoning- Clostridium botulinum

 Pseudomembrane colitis- Clostridium difficile


MORPHOLOGY
 Highly pleomorphic
 3-8 µ -0.4-1.2 µ
 Gram variability
 Motile with peritrichate flagella (Except Clostridium perfringens and
Clostridium tetani Type IV))
 C.perfringens/C. butyricum capsulated
SPORES
Generally subterminal except in-
CULTURE
 Exacting anaerobes to aerotolerant
 Anaerobic media- reducing substances
 Unsaturated fatty acids
 Ascorbic acid
 Glutathione
 Cysteine
 Thioglycolic acid
 Metallic iron
 Alkaline glucose

 Robertson cooked meat media


RESISTANCE
 Vegetative cells similar to other bacteria
 Spores variable resistance
 Resistant to phenolic disinfectants
 Glutaraldehyde very effective
PATHOGENECITY
 Invasive powers limited
 Exotoxin production
 C.perfringens can also be invasive
CLOSTRIDIUM PERFRINGENS
 C.welchii, Bacillus aerogenes capsulatus. B.phlegmonis emphysematosae

 Plump GPB

 Capsulated, non motile


 Subterminal bulging spores

 Invasive as well as toxigenic

 Commensal of large intestine of humans and animals

 Most important cause of gas gangrene also causes food poisoning,


necrotising enteritis
CULTURAL
CHARACTERISTICS
 Anaerobe but can grow in microaerophilic conditions
 Ph- 5.5- 8
 Optimum 45 o C
 Saccharolytic
 Stormy clot reaction in Litmus milk
VIRULENCE FACTORS
 C.perfringens classified into 5 strains A- E based on major toxin
production
 Major toxins- Alpha, beta, epsilon and iota
 Minor toxins- eight (γ , 𝛿 , ƛ , 𝜅 , θ , μ etc)
 Enterotoxins

 Alpha (⍺ )
 All strains maximally by Type A
 Responsible for gas gangrene in humans
 Lethal ,dermonecrotic, haemolytic
 Phospholipidase- Lecithinase C splits lecitihine to phosphoryl choline and
diglyceride
NAGLER REACTION
 Media- 6 % agar , 5 % Filde’s digest of sheep blood and 20% human
serum/ 5% egg yolk
 ½plate- antitoxin
 Opacity surrounding the streak line due to lecithinase activity of alfa
toxin
 No opacity on side with the antitoxin
PATHOGENECITY
 Anaerobic wound infections-
 Simple wound contamination- Delayed wound healing
 Anaerobic cellulitis- Fascial planes- Gas Abscess
 Anaerobic myositis- Gas gangrene
PATHOGENECITY
 Food poisoning
 Heat labile enterotoxin
 Cold /warmed meat dish
 Incubation period 8- 24 hours self limiting

 Gangrenous appendicitis
 Necrotising enteritis
 Brain abscesses
 Panophthalmitis
GAS GANGRENE
 Malignant edema or Clostridial myonecrosis
 Rapidly spreading, edematous myonecrosis –Usually in association with
severe wounds with extensive muscle damage contaminated with
pathogenic clostridia
 War
 RTA
 Organisms associated- polymicrobial
 C.perfringens- 60 %
 C.novyi and C.septicum -20-40%
 C.histolyticum
GAS GANGRENE
 Conditions favourable for clostridial multiplication
 Low O2 tension
 Crush injuries /arterial disruption-Anoxia
 Extravasation-Reduces blood flow
 Decreased Eh and pH
 Extravasated Haemoglobin and myoglobin reduced
 Anaerobic reduction of pyruvate to lactate – Further fall in Eh
GAS GANGRENE
 Clostridial multiplication – Toxin production ( ⍺ and θ)

 Lecitihinases- Cell membrane damage and capillary damage


 Haemolysis and haemoglobinuria
 Collagenases and hyaluronidases
 Abundant gas
GAS GANGRENE
 Clinical presentation
 Incubation period varied average 10-48 hours
 Thin watery wound discharge – Becomes serosanguinous and profuse
 Crepitant
 Profound toxemia and prostration
LABORATORY DIAGNOSIS
 Clinical
 Laboratory- to confirm clinical diagnosis and species identification
 D/D
 Streptococcal myositis- Gram stain – Numerous pus cells and Gram +ve
cocci
SPECIMENS
 Smears from the muscle at the edge of the affected area
 Exudates from depths of the wound
 Necrotic tissue and muscle fragments
 Blood culture
 Swabs- not to be taken

 Specimen inoculated in RCM and taken to laboratory


GRAM STAIN
 Absence of neutrophils
 Polymicrobial-
 C. perfringens - Large numbers of regularly shaped bacilli without spores
 C. septicum - Boat or leaf shaped pleomorphic bacilli with “citron bodies”
 C.novyi – Large bacilli with oval subterminal spores
CULTURE
 Aerobic and anaerobic culture
 Blood agar- target hemolysis
 Inner narrow zone of complete hemolysis- θ toxin
 Wider zone of incomplete hemolysis- ⍺ toxin)

 Serum/ egg yolk agar -Nagler reaction


REVERSE CAMP TEST
 To differentiate Clostridium perfringens from
other Clostridium species
 Group B Streptococcus streaked in centre
 Test organism streaked perpendicular to it
 ⍺ toxin produced by C perfringens interacts with
CAMP factor and produce synergistic hemolysis
 “Arrowhead” hemolysis
TREATMENT
 Surgery

 Hyperbaric O 2

 Antibiotics – Metronidazole
 Broad spectrum cover
 Passive immunization Polyvalent antitoxin
CLOSTRIDIUM BOTULINUM
 Botulism
 Saprophyte
 Gram +ve 5 x 1 µ
 Subterminal, oval, bulging spores
CULTURAL
CHARACTERISITICS
 Strict anaerobe
 Optimum 35 o C
 Heat resistant spores
 Classification- Eight serotypes (A- G)
TOXIN
 Powerful neurotoxin- Botulinum toxin
 Produced intracellularly (not secreted)
 Liberated on cell death
 MW- 70,000
 LD for mice 0.000,000,033 mg, Humans 1-2 µg
 Pressure cook/ boil for 20 mins – Toxin destroyed
 Action- Blocks release of Acetyl choline at synapses and neuromuscular
junctions
 Flaccid paralysis
TOXIN
 Diplopia, dysphagia and dysarthria
 Symmetric descending paralysis
 Death by respiratory paralysis
 Small quantity injected – Muscles atrophy –Recover after 2- 4 months
restore transmission
BOTULISM
 Ingestion of preformed toxin
 Human infections A, B , E and F
 Canned food – Bubbles on opening
 Vomiting , thirst, constipation, difficulty in swallowing , coma and
delirium.
 Descending symmetric flaccid paralysis
 No sensory deficit
 Respiratory muscle paralysis- death
BOTULISM
 Wound botulism- Wound infection
 Infant botulism
 Wild honey
 Below 6 months of age
 Constipation, weakness floppy movements loss of head control
LABORATORY DIAGNOSIS
 Bacillus/ toxin in food / feces
 Food macerated in sterile saline and filtrate inoculated into mice
 Typing by specific antitoxin
 Retrospective – Antitoxin in serum
 Treatment - Antitoxin
CLOSTRIDIUM TETANI
 GPB

 Terminal spherical spores- drum stick


appearance

 Tetanus- skeletal muscle spasm and


autonomic nervous system disturbance

 Present in soil and intestines of humans and


animals
TOXINS

Produce exotoxin- Tetanospasmin


Heat labile, 150 KDa peptide
Plasmid coded
Neurotoxin- binds to receptors on motor nerve terminals
Prevent presynaptic release of inhibitory neurotransmitters (Glycine
and GABA)
Cause spastic muscle contraction
MODE OF TRANSMISSION
 Injuries
 Surgery without proper asepsis
 Unhygenic practices during abortions/ delivery
TETANUS INTOXICATION
Entry wound
Spread
blood stream
moves up nerves
Disease
Incubation: 6-10 days
Muscles of face and jaw affected first- Lock jaw
Painful muscle spasm
Descending spastic paralysis
cardiac failure, respiratory failure
LOCKJAW
TETANUS
Trismus
Risus sardonicus- spastic contraction of facial
muscles
Opisthotonos- generalized spastic contraction of
extensor muscles
Localized -- favourable prognosis
Prevention: toxoid, 3% formaldehyde
LAB DIAGNOSIS
 Specimen- excised tissue bits from necrotic wound
 Gram stain- GPB with drum stick appearance
 Culture- produce swarming growth in blood agar with polymyxin B
PREVENTION
 Active Immunization
 Tetanus toxoid
 Given under national immunization schedule- 3 doses at 6,10 and 14 weeks
and 2 booster doses at 16-24 weeks and 5 years

 Prevention of tetanus after injury


 Surgical toilet with immunization

 Passive Immunization
 Tetanus immunoglobulin given as treatment of choice for tetanus

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