CLOSTRIDIUM

GRAM POSITIVE BACILLI

F

Clostridium are strict anaerobe (but some are aerotolerant)

Clostridium are strict aerobe (t/f)

Clostridium spp.

It is frequently encountered in exogenous anaerobic infections or intoxications

Catalse –

Clostridium is Catalse + or –

1. Collagenase

2. Hyaluronidase

3. Phopholipase

4. Lecithinase

Clostridium

Enzymes that contribute to the virulence:

Clostridium sordelii

only urease (+) clostridia

Clostridium ramosum

Clostridium inoculum

Clostridium perfringens

RIP

Clostridium

All are motile except

Clostridium bifermentans

Clostridium perfringens

BP

Clostridium

All are with swollen spores except

Clostridium perfringens

Clostridium

All are non-capsulated except

Clostridium perfringens

Clostridium septicum

PS

Clostridium

All are lactose (-) except

Clostridium perfringens

Clostridium

All are sucrose (-) except

Clostridium perfringens

Clostridium

All are single hemolytic except

Clostridium tetani

Clostridium

All are saccharolytic except

Clostridium histolyticum

Clostridium tetani

HT

Clostridium

All are dextrose (+) except

1. Peptone-Yeast Glucose Broth (PYG)

2. Egg Yolk Agar

3. Cycloserine Cefoxitin Fructose Agar

4. Transport medium

   PECT

Media for Isolation of Clostridium

Egg Yolk Agar

to detect the activities of Lipase and Lecithinase

Peptone-Yeast Glucose Broth (PYG)

detects volatile fatty acids

Cycloserine Cefoxitin Fructose Agar

for C. difficile detection only

Transport medium

PRAS (Pre-reduced Anaerobically Sterilized) and AMIES

Neurotoxic (brain)

1. C. tetani

2. C. botulinum

Histotoxic (tx)

1. C. bifermentans

2. C. perfringens

3. C. septicum

4. C. novyi

5. C. chauvoei

Enteric

1. C. difficile

Types of Clostridium

Frankel’s / Gas Gangrene Bacillus

Clostridium perfringens / welchii is commonly known as

F

Clostridium perfringens / welchii is anaerobic / encapsulated, non-motile double / alpha-prime hemolysis

Clostridium perfringens / welchii is aerobic / capsulated, non-motile double / alpha-prime hemolysis (t/f)

Boxcar-shaped bacillus

Clostridium perfringens / welchii morphology is

wound contact with soil

Clostridium perfringens / welchii source is

subterminal spores

Clostridium perfringens / welchii is round with what kind of spores

1. Beta-hemolysin & Theta Toxin

2. Alpha-Toxin

   • Lecithinase (Phospholipase)

   • causes Gas Gangrene

Virulence Factor of Clostridium perfringens / welchii

1. Gas Gangrene

2. Food Poisoning

   1. Type A

   2. Type C

3. Necrotic Enteritis

   1. Pig Bel

Diseases of Clostridium perfringens / welchii

Myonecrosis

Diseases of Clostridium perfringens / welchii

Gas Gangrene

Food poisoning

Diseases of Clostridium perfringens / welchii

enterotoxins

1. Type A

2. Type C

Diseases of Clostridium perfringens / welchii

Types of Food poisoning

Type A

Diseases of Clostridium perfringens / welchii

Type of Food poisoning that is mild & self-limiting

Type C

Diseases of Clostridium perfringens / welchii

Type of Food poisoning that is serious but rarely encountered

Type C

Diseases of Clostridium perfringens / welchii

Type of Food poisoning that experiences bloody diarrhea and vomiting

Pig Bel

Diseases of Clostridium perfringens / welchii

Type of Necrotic Enteritis that is a form of bloody diarrhea

1. Chopped Meat Agar

2. Reverse Camp Test

3. BAP

4. Nagler’s Test / Lecithinase

5. Stormy Fermentation of Milk

Laboratory Tests for Identification of Clostridium perfringens / welchii

Chopped Meat Agar

Laboratory Tests for Identification of Clostridium perfringens / welchii

Growth + gas ; anaerobic growth

S. agalactiae

Laboratory Tests for Identification of Clostridium perfringens / welchii

Known for Reverse Camp Test

C. perfringens / welchii

Laboratory Tests for Identification of Clostridium perfringens / welchii

Unknown for Reverse Camp Test

Arrowhead zone of beta-hemolysis

Laboratory Tests for Identification of Clostridium perfringens / welchii

+for Reverse Camp Test

Double zone / Target / Alpha Prime Hemolysis

Laboratory Tests for Identification of Clostridium perfringens / welchii

BAP

Beta-hemolysis (Theta Toxin)

Laboratory Tests for Identification of Clostridium perfringens / welchii

In of BAP

Alpha-Hemolysis

Laboratory Tests for Identification of Clostridium perfringens / welchii

Out of BAP

1. Lecithinase

2. alpha toxin

3. phospholipase C

LAP

Laboratory Tests for Identification of Clostridium perfringens / welchii

Nagler’s Test / Lecithinase is due to

McClung / Neomycin Egg Yolk Agar

Laboratory Tests for Identification of Clostridium perfringens / welchii

Nagler’s Test / Lecithinase’s Media is

Opalescence of agar without anti-toxin; inhibition of lecithinase reaction

Laboratory Tests for Identification of Clostridium perfringens / welchii

+McClung / Neomycin Egg Yolk Agar

NO OPALESCENCE on agar with anti-toxin

Laboratory Tests for Identification of Clostridium perfringens / welchii

– McClung / Neomycin Egg Yolk Agar

F

Stormy Fermentation of Milk is + for coagulase casein/clotting of protein + gas

Laboratory Tests for Identification of Clostridium perfringens / welchii

Stormy Fermentation of Milk is – for coagulase casein/clotting of protein + gas (t/f)

PINK

Laboratory Tests for Identification of Clostridium perfringens / welchii

Result of Stormy Fermentation of Milk for Acid

BLUE

Laboratory Tests for Identification of Clostridium perfringens / welchii

Result of Stormy Fermentation of Milk for Alkaline

Clostridium tetani

Iron-Loving Bacteria

3 - 21 days

Clostridium tetani’s incubation period

• Tetanospasmin

• Tetanolysin

Virulence Factors (Exotoxin) of Clostridium tetani

Tetanospasmin

– an exotoxin binds to ganglioside receptors which block the release of NEUROTRANSMITTER & inhibits neurons in CNS;

Tetanospasmin

causes SPASTIC PARALYSIS

Tetanolysin

– causes beta-hemolysin

• Spastic Paralysis

• Tetanus

• Risus Sardonicus / Sardonic Smile

• Opisthotonus

• Tetanus Neonatorum

Diseases Associated of Clostridium tetani

Spastic Paralysis

Diseases Associated of Clostridium tetani

– with little movement

Tetanus

Diseases Associated of Clostridium tetani

– lock jaw / “trismus”

Opisthotonus

Diseases Associated of Clostridium tetani

– arching of the back

Tetanus Neonatorum

Diseases Associated of Clostridium tetani

– contaminated instrument used for newborns

Administration of anti-toxin

Treatment of Clostridium tetani

Tetanus toxoid

Prevention of Clostridium tetani

• Clinical Findings – basis for diagnosis

• Morphology Test

• Non-saccharolytic

Laboratory Test for ID of Clostridium tetani

Clinical Findings

Laboratory Test for ID of Clostridium tetani

– basis for diagnosis

- Terminal oval spore

- Tennis racket, drumstick

- Tack head bacillus

Laboratory Test for ID of Clostridium tetani

Morphology Test - Gram stain shows

Clostridium botulinum

Von Emergen’s / Canned Goods Bacillus

F

Clostridium botulinum is NOT CULTURED

Clostridium botulinum is cultured (t/f)

Botulinum Toxin

Virulence Factors of Clostridium botulinum

a neurotoxin that blocks the release of ACETYLCHOLINE

Acetylcholine

Virulence Factors of Clostridium botulinum

Botulinum Toxin is a neurotoxin that blocks the release of

Botulinum toxin

Virulence Factors of Clostridium botulinum

the most potent exotoxin

- 7 Types: A, B, C, D, E, F, G

Virulence Factors of Clostridium botulinum

Botulinum Toxins has how many types and what are they

A, B, & E

Virulence Factors of Clostridium botulinum

Type of Botulinum toxins that is associated with human infection

• Flaccid Paralysis

• Food Botulism

• Wound Botulism

• Infant Botulism

• Sudden Infant Death Syndrome (SIDS) / Crib Death

Diseases Associated with Clostridium botulinum

Flaccid Paralysis

Diseases Associated with Clostridium botulinum

no movement

Food Botulism

Diseases Associated with Clostridium botulinum

home-canned goods with preformed toxin

Wound Botulism

Diseases Associated with Clostridium botulinum

spore on wound

Diseases Associated with Clostridium botulinum

spore ingestion via breast via breastfeeding

Infant Botulism

Diseases Associated with Clostridium botulinum

reduced muscle strength (hypotonia)

hypotonia

Diseases Associated with Clostridium botulinum

Infant Botulism causes reduced muscle strength or aka

Infant Botulism

Diseases Associated with Clostridium botulinum

- FLOPPY BABY SYNDROME

• BAP

• + Lipase reaction on Egg Yolk Agar

• – Lecithinase & Indole

• Ferments Glucose

• Oval Subterminal Spores

• Mouse Neutralization Test

Lab Test for ID of Clostridium botulinu

+Lipase reaction on Egg Yolk Agar

Lab Test for ID of Clostridium botulinum

Lipase reaction on Egg Yolk Agar

– Lecithinase & Indole

Lab Test for ID of Clostridium botulinum

Lecithinase & Indole

Glucose

Lab Test for ID of Clostridium botulinum

Ferments -

Oval Subterminal Spores

Lab Test for ID of Clostridium botulinum

Spores

beta-hemolysis

Lab Test for ID of Clostridium botulinum

BAP

Mouse Neutralization Test

Lab Test for ID of Clostridium botulinum

Definitive ID test for C. botulinum

Mouse Neutralization Test

Lab Test for ID of Clostridium botulinum

Detects the presence of neurotoxin in serum, feces, gastric contents, vomitus, or food.

F

Mouse Neutralization Test detects the presence of neurotoxin in serum, feces, gastric contents, vomitus, or food.

Lab Test for ID of Clostridium botulinum

Mouse Neutralization Test detects the presence of enteric toxin in serum, feces, gastric contents, vomitus, or food. (t/f)

Clostridium difficile

Normal flora of the COLON

Clostridium difficile

Major cause of HOSPITAL DIARRHEA

• Toxin A : Enterotoxin

• Toxin B: Cytotoxin

Virulence Factor of Clostridium difficile

Enterotoxin

Virulence Factor of Clostridium difficile

Toxin A

Cytotoxin

Virulence Factor of Clostridium difficile

Toxin B

Antibiotic-Associate Pseudomembranous Enterocolitis

Diseases Associated with Clostridium difficile

- Found in px who have received one or more broad-spectrum antibiotics

Antibiotic-Associate Pseudomembranous Enterocolitis

Diseases Associated with Clostridium difficile

- Produces toxins, leading to diarrhea

diarrhea

Diseases Associated with Clostridium difficile

Antibiotic-Associate Pseudomembranous Enterocolitis produces toxins, leading to -

Antibiotic-Associate Pseudomembranous Enterocolitis

Diseases Associated with Clostridium difficile

- Produces GLUTAMATE DEHYDROGENASE for determination of C. difficile toxin

GLUTAMATE DEHYDROGENASE

Diseases Associated with Clostridium difficile

Antibiotic-Associate Pseudomembranous Enterocolitis produces - for determination of C. difficile toxin

• BAP

• Oval Subterminal Spore

• Direct Detection of toxin from the stool by enzyme immunoassay (EIA)

• Tissue/Cell Culture/ Cytotoxin Assay

• Does not ferment lactose, maltose, xylose but FERMENTS FRUCTOSE

• + Gelatinase

• + Cycloserine Cefoxitin Fructose Agar

• – Lipase, Lecithinase, Indole

Laboratory Test for ID for Clostridium difficile

fluoresces chartreuse; yellow fluorescence; beta-hemolytic

Laboratory Test for ID for Clostridium difficile

BAP