lectures 8-11-bacillus anthracis; b. cereus, clostridium perfringens, cl. tetani; cl. botulinum; cl....
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بسم الله الرحمن الرحيمبسم الله الرحمن الرحيم
FAMILY: BACILLIACEAEFAMILY: BACILLIACEAEProf. Khalifa Sifaw GhengheshProf. Khalifa Sifaw Ghenghesh
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1. GENUS: 1. GENUS: BACILLUSBACILLUS
• Gram +ve bacilli
• Aerobic
• Spore-Forming
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i. i. Bacillus anthracisBacillus anthracis• >> Anthrax.
• Large, Square - ended Rods, Arranged in Chains.
• Non-Motile.• Spores:• Capsule: – Purple Stained >> McFadyan's Method
(Polychrome Methylene Blue).
• Colonies on BA: "Medusa Head Appearance"
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Bacillus anthracisBacillus anthracis
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An electron micrograph of spores from An electron micrograph of spores from the Sterne strain ofthe Sterne strain of Bacillus anthracisBacillus anthracis
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Bacillus anthracisBacillus anthracis McFaydean capsule McFaydean capsule stain, grown at 35stain, grown at 35ooC, in defibrinated C, in defibrinated
horse blood.horse blood.
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DISEASE:• In Animals: >> Septicaemia.
• In Humans: i. Cutaneous Anthrax > Malignant pustule
ii. Pulmonary Anthrax (Wool-Sorter'sDisease).
iii. Gastrointestinal Anthrax.
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Cutaneous AnthraxCutaneous Anthrax
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Anthrax lesion on the skin of the Anthrax lesion on the skin of the forearm caused by forearm caused by Bacillus anthracisBacillus anthracis
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PATHOGENESIS• Capsule > Invasiveness– D-glutamic acid
• Exotoxin (Plasmid mediated)i. Protective Factor (Antigen).
ii. Oedema Factor.
iii. Lethal Factor.
Blocks the Adenyl Cyclase Pathway > Increases vascular Permeability > Shock
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LABORATORY DIAGNOSIS:• Specimens obtained from:
a malignant pustule, sputum, blood.
- Gram stain + fluorescent-antibody stain.
- Motility
- Capsule formation: Sodium bicarbonate +CO2
- String-of-pearls reaction:
- Mouse test:
- API
>> Demonstration of Abs to the organism:
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Bicarbonate agar and blood agar Bicarbonate agar and blood agar plate cultures of plate cultures of Bacillus anthracisBacillus anthracis
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Negative encapsulation: Blood agar and Negative encapsulation: Blood agar and bicarbonate agar plate cultures of bicarbonate agar plate cultures of
Bacillus cereusBacillus cereus
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• TREATMENT– Penicillin, Ciprofloxacin
• IMMUNIZATION–Animals > Live spore vaccine
(Sterne strain)
–Workers at Risk of Exposure >
Anthrax Vaccine Absorbed (AVA) >> “Alum precipitated toxoid”
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ii. ii. Bacillus cereusBacillus cereus
• Food Poisoning.
• Clinical Syndromes:
i. Severe Nausea &Vomiting.
ii. Abdominal Cramps & Diarrhoea.
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PATHOGENICITY:>> Due to an Enterotoxin.
• Also Causes Disease in Patients with Underlying Disease.
• TREATMENT:
>> Tetracycline, Erythromycin.
• iii. B. subtilis:
• iv. B. stearothermophilus.
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2. GENUS: 2. GENUS: CLOSTRIDIUMCLOSTRIDIUM
• Gram +ve bacilli
• Anaerobic,
• Spore Forming
- Spores:
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Ink Stain of Sporulating Ink Stain of Sporulating ClostridiumClostridium--spores appear clear, vegetative cells darkspores appear clear, vegetative cells dark
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i.i. Clostridium perfringens Clostridium perfringens
• Nonmotile
• Spores Not Produced in Ordinary Media.
• Aerotolerant Anaerobe.
• 5 Types: A - E
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Gram stain of Gram stain of Clostridium perfringensClostridium perfringens
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Exudate smear of Exudate smear of Clostridium perfringensClostridium perfringens
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Tissue smear of Tissue smear of Clostridium perfringensClostridium perfringens
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DISEASE:
• Clostridial Myonecrosis.
• Less Severe Wound Infections.
• Food Poisoning.
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Patient with gas gangrenePatient with gas gangrene
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LABORATORY IDENTIFICATION
• In Chopped Meat - Glucose Medium:
• On BA:
• On Egg Yolk Agar: >> Precipitation (Opalescence).
• Milk Media: Stormy Formation.
• Nagler Reacrion:
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Blood agar plate with Blood agar plate with Cl. perfringensCl. perfringens characteristic double zone of hemolysischaracteristic double zone of hemolysis
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PATHOGENICITY & CLINICAL INFECTION-Toxin: Acts on Lecithin-Containing Lipo-
protein Complexes in the Cell Membrane.
• Predisposing Factors:i. Trauma with Deep and Lacerated or Crush
Wounds of Muscle Etc.
ii. Require a Reduced Oxygen Tension and
Reduced Oxidation Reduction Potential
for Growth.
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FOOD POISONING:
• Cl. perfringens Type A >> Enterotoxin.
> Acute Abdominal Pain and Diarrhoea.
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LABORATORY DIAGNOSIS:• Important: Diagnosis of Clostridium
Myonecrosis Should Be Rapid and Made on Clinical Grounds.
i. Direct Smear and Gram Stain of Material
from Deep Within the Wound.
ii. Culture: Tissue Aspirates or Deep Swabs Taken from Affected Muscle.
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TREATMENT:• Clostridium Myonecrosis:
i. Surgical Removal of All Infected and
Necrotic Tissue.
ii. Antibiotic and Antitoxin Therapy.
iii. Adminstration of Hyperbaric Oxygen.
• Food Poisoning:
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Clostridia That May Be Associated
with Gas Gangrene:
• Cl. perfringens Type A• Cl. septicum• Cl. novyi Type A• Cl. histolyticum• Cl. Sordellii
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Human case of malignant edema Human case of malignant edema caused by caused by Cl. septicumCl. septicum
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ii. ii. Clostridium tetaniClostridium tetani
• > Tetanus.
• > Terminal Spores with Drumstick
Appearance.
• > Obligate Anaerobe.
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Clostridium tetaniClostridium tetani Gram Positive RodsGram Positive Rods
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Clostridium tetaniClostridium tetani
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VIRULENCE FACTORS:• Tetanus Toxin (Tetanospasmin) >
Neurotoxin.
i. An Intercellular Toxin Released by
Cellular Autolysis.
ii. Inhibits the Release of Inhibitory
Transmitters.
iii. Toxoid.
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CLINICAL INFECTION & PATHOGENESIS• "Tetanus is Generalized in Nature".
i. Unimmunized Rural Population.
ii. In Practice: Simple Puncture Wounds >
Nail, Splinter or Thorn.
iii. In Traumatic Wounds > Compound
Fractures, Dental Extractions, Etc.
iv. Tetanus Neonatrum:
v. Postoperative Tetanus:
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Drawing of a Soldier dying of Drawing of a Soldier dying of Tetanus (Opisthotonos)Tetanus (Opisthotonos)
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A patient presented with facial tetany. A patient presented with facial tetany. Note the contraction of the masseter and Note the contraction of the masseter and
neck musclesneck muscles
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LABORATORY DIAGNOSIS:• > Diagnosis on Clinical Grounds.
TREATMENT:• i. Antitoxin.• ii. Debridement of Wound and Removal of • any Foreign Bodies.• iii. Pencillin >>> In Large Doses.• iv. Mild Tetanospasm: >>> Barbiturates.• v. Severe Cases: • >>> Use Curare - Like Agents.• >>> Tracheostomy.• >>> Careful Control of the
Environment.
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PREVENTION:> Prompt and Adequate Cleaning of
Wounds.
i. Active Immunity.
ii. Passive Immunity.
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iii. iii. Clostridium botulinumClostridium botulinum
• > Botulism.
• > Gram +ve, Spore Forming Bacilli.
• > Strict Anaerobe.
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Gram Stain of Gram Stain of Cl. botulinumCl. botulinum, , Characteristic Long RodsCharacteristic Long Rods
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A photomicrograph of A photomicrograph of Clostridium botulinumClostridium botulinum type A type A
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Blood Agar Plate with Blood Agar Plate with C. botulinumC. botulinum
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VIRULENCE FACTORS• Botulinum Toxin >>> Neurotoxin.–Serologically 8 Toxins >>
A, B, C1, C2, D, E, F & G.
> Affect the Cholinergic System > Blocks the Release of Acetylcholine (at Points in Peripheral Nervous System).
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DISEASE IN HUMANS1. Food - Borne Botulism:> Incubation Period: 12-36 Hours to 8 days.
2. Infant Botulism:
LABORATORY DIAGNOSISi. Diagnosis Made Clinically.ii. Detection of Organism or Its Toxin in the Suspected Foodiii. Samples of Stool or Vomit
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TREATMENT & PREVENTIONImportant: Specific Treatment Should
Begin as Quick as Possible.
>Polyvalent Antitoxin >>> Immediately.
>Physiological Support >>> ICU.
>NEVER Use a Swollen or Defective Can.
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iv. iv. Clostridium difficileClostridium difficile
• Antibiotic Associated Colitis.
• Produce Two Major Protein Toxins
(A &B).
• Risk Factors: –Antibiotic Exposure.
–Old Age.
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Clostridium difficileClostridium difficile
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Scanning electron micrograph of Scanning electron micrograph of Clostridium difficleClostridium difficle
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Intestinal Smear- Close Association Intestinal Smear- Close Association of of Cl. difficileCl. difficile with Neutrophils with Neutrophils
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• Infection Can Be: –Endogenous or Exogenous.
• Nosocomial Spread: Due to Spores.
LAB DIAGNOSIS:1. Demonstration of Cytotoxin in Stool.
2. Isolation of the Microorganism.
TREATMENT:–Discontinuing Treatment.
–Vancomycin.