CLOSTRIDIUM

G+ve, anaerobic, spore forming bacilli.

Disease gas gangrene, food poisoning & tetanus
Pleomorphic
Pathogenic Cl.welchii & Cl.tetani
Industrially important Cl.acetobutylicum acetone
& butanol.
Motile & peritrichate
Cl.welchii & Cl.tetani non motile
Opt T -37 C, PH 7 7.4
CHARACTERISTIC Cl.perfringes/ Cl.tetani Cl.botulinum
Cl.welchii

General Character Non motile Motile polar flagella Motile peritichate

Capsulated Non capsuated Non capsulated
Spore Spore terminal Spore-subterminal
central/subterminal bulging drum stick bulging spore,
appearance
Cultural Anaerobic/ Obligate anaerobe Strict anaerobe
Characteristics microaerophilic PH 7.4 PH 7.4
PH 5.5 8.0 Temp 37 C Opt Temp 35 C
Opt Temp 37 C/45 Spores 20-25 C in
C alkaline glucose
gelatin media

Growth media Enrichment media Blood agar buffy Ordinary media

Robertson cooked ball Spores - alkaline
meat broth Gelatin deep fir tree glucose gelatin media
Litmus milk-stormy appearance
fermentation , acid Mac conkey agar
clot &gas greenish fluoroscent,
non lactose fermenters
CHARACTERISTIC Cl.perfringes/ Cl.tetani Cl.botulinum
Cl.welchii

Growth media Blood agar-Target Blood agar &

haemolysis, double haemolysis
zone pattern. First Robertson cooked
complete haemolysis meat broth turbidity
theta toxin with gas
Second incomplete
haemolysis alpha
toxin
Biochemical test Ferment sugar acid Do not ferment
& gas
IMV -+- IMViC -++-
H2S produced H2S not produced
Nitrates reduced Nitrates not reduced

Classification Based on toxin Based on serotyping Based on

production, 5 types 1-10 types, type 6 non immunological
A-E classes flagellated difference in toxins, 8
types, A - H
CHARACTERISTIC Cl.perfringes/ Cl.tetani Cl.botulinum
Cl.welchii

Toxins toxin Tetanolysin Exotoxin

Phospholipase(lecithin haemolysin pathogenicity
ase C) produced by all Tetanospasmin Produced
strain, abundantly by neurotoxin intracellularly
type A. Unknown toxin non appears on medium
Toxemia & gas spasmogenic toxin autolysis
gangrene Tetanolysin O2 & Non toxic(initially)
diglyceride heat labile, leucotoxin activated by trypsin
Lecithin Mg2+,Cs2+ Tetanospasmin Neurotoxin acts
highly toxic, tetanus slowly
phosphorylcholine Blocks release of
toxin lethal, acetyl choline at
dermonecrotic & synapses/neuromuscul
hemolytic ar junctions
Dysphagia, respiratory
paralyses, death due to
respiratory failure
CHARACTERISTIC Cl.perfringes/ Cl.tetani Cl.botulinum
Cl.welchii

Naglers reaction:
Media 6%
agar+5% fildes
peptic digest of sheep
blood + 20% human
serum

Plate divided into 2

halves

One half added with

specific antisera

Organism inoculated
into both halves

Opacity produced in
the half without
antisera due to
lecithinase enzyme
CHARACT Cl.perfringes/ Cl.welchii Cl.tetani Cl.botulinum
ERISTIC

Hot Cold Phenomenon

toxin in blood agar

Incubate at 37 C

Chilled at 4 C

Visualize haemolysis
Beta (), epsilon (), iota ()
lethal & necrotising
Eta () hemolytic
Delta () hemolytic
Theta () O2 labile hemolysin
Kappa () collagenase
Lambda () proteinase &
gelatinase
Mu () hyaluronidase
Nu () - deoxyribonuclease
CHARACTERISTIC Cl.perfringes/ Cl.tetani Cl.botulinum
Cl.welchii
Pathogenesis Produce no of Cl. tetani spores in Non invasive & non
infections in human subcutaneous wound infectious
Gas Gangrene Action of toxin
Occur through a Germination of spores botulism
wound infection. (6-12 days) foodborne
Characterized by Botulism wound
rapidly spreading Produce toxin infant
edema,myositis,necro Food Borne
sis of tissue with gas Toxin absorbed by Ingestion of toxin A,B
production. motor nerve endings E & rarely F
Leads to multiorgan C & D wild fowl &
failure, coma and Transported thro alpha cattle
death. fibres of motor nerves E fish & sea food
Food Poisoning Cans inflated,
Strain A alpha or Reach CNS bubble on opening
theta toxin Characterized by
Abdominal Toxin fixed to vomiting, thirst,
pain,diarrhoea,vomiti ganglioside grey constipation, difficulty
ng after 8-24 hrs matter of nerve tissues in swallowing,
Self limiting after 24- speaking & breathing
48 hrs. Coma & death
respiratory failure
1-7 days after onset
CHARAC Cl.perfringes/ Cl.welchii Cl.tetani Cl.botulinum
TERISTI
C
Toxin inhibits the synaptic Wound botulism
inhibition in the spinal cord Rare, toxin site of
infection and is absorbed
Affect the terminals that use Symptoms similar to food
glycine as neurotransmitter
borne botulism
Abolish the spinal inhibition Infant botulism
control Spores ingested
established in gut &
Spread of controlled produce toxin
impulse throughout CNS Infants below 6 months
Constipation, poor
Stimulate simultaneous feeding, lethrgy,
contraction of agonistic &
antagonists
weakness, pooled oral
secretion, weak or altered
Muscular rigidity & spasm cry, loss of head control
Honey likely food
Lockjaw, pain, stiffness of bacillus enters gut.
neck
CHARACTER Cl.perfringes/ Cl.welchii Cl.tetani Cl.botulinum
ISTICl
Gangrenous appendicitis
Strain A and D (toxin)
Cause intestine obstruction &
peritonitis
Necrotising enteritis
Strain C
Toxin haemolysis
Contaminated pork
Fatal necrotising jejunitis
Bilary tract infection
Acute emphysematous
cholecystitis
Post chole cystectomy septicemis
Brain abscess
Meningitis
Spreads in CSF
Pan ophthalmitis
Penetration and infection in eye
Thoracic infection
Organism penetrates and infects
from chest cavity to thorax
Common in battlefields
Urogenital infection
Infection in urinary tract
Occurs through surgical
procedure
Septicemia is common
CHARACTERISTIC Cl.perfringes/ Cl.tetani Cl.botulinum
Cl.welchii

Lab diagnosis
a. Necrotic tissue in the a.Wound exudate/ necrotic Suspected food, feces
Specimen wound tissue
b. Suspected food b. Wound swabs
c. Feces c. Infected muscle fragment
d. CSF
e. Urine
Microscopy G +Ve bacilli G +Ve bacilli with spores as G +Ve sporing bacilli
drum stick appearance

Culturing
1.Specimen Robertsons Aerobic & anaerobic Food macerated with sterile
cooked meat broth culture methods saline
Inoculated at 37C for 4-6 6% agar in horse blood agar
hrs or heated blood agar with Filtrate inoculated in
Subcultured in blood agar antitoxin mice/guinea pig
intraperitoneally

Control protected by
polyvalent antitoxin
CHARACTERISTIC Cl.perfringes/ Cl.tetani Cl.botulinum
Cl.welchii
Prophylaxis Wounds cleaned with Three methods Antitoxin intramuscularly
antiseptics povidone- 1.Surgical attention Active immunisation
iodine Remove foreign effective
bodies,necrotic tissue and Injection of AlSO4 adsorbed
blood clot to avoid toxoid given at interval of
anaerobic condition 10 weeks followed by
Antiseptic povidone iodine booster dose
applied Combination of guanidine
2.Antibiotic prophylaxis HCl & germine
Long acting penicillin, monoacetate administered
erythromycin 500mg,
neomycin local application
3.Immunization
Passive immunization
1500 IU of antitetanus
serum subcutaneously
Human antitetanus
immunoglobulin to avoid
hypersensitivity
Active immunization
DPT-triple vaccine in 3
doses with interval of 4-6
weeks between first two
injection