‫مممممم‬ ‫مممممم‬ ‫مممم‬ ‫ممم‬‫مممممم‬ ‫مممممم‬ ‫مممم‬ ‫ممم‬
FAMILY: BACILLACEAEFAMILY: BACILLACEAE
Prof. Khalifa Sifaw Gh...
1. GENUS:1. GENUS: BACILLUSBACILLUS
• Gram +ve bacilli
• Aerobic
• Spore-Forming
i.i. Bacillus anthracisBacillus anthracis
• >> Anthrax.
• Large, Square - ended Rods, Arranged in
Chains.
• Non-Motile.
• ...
Bacillus anthracisBacillus anthracis
An electron micrograph of spores fromAn electron micrograph of spores from
the Sterne strain ofthe Sterne strain of Bacill...
Bacillus anthracisBacillus anthracis McFaydean capsuleMcFaydean capsule
stain, grown at 35stain, grown at 35oo
C, in defib...
DISEASE:
• In Animals: >> Septicaemia.
• In Humans:
i. Cutaneous Anthrax > Malignant pustule
ii. Pulmonary Anthrax (Wool-S...
Cutaneous AnthraxCutaneous Anthrax
Anthrax lesion on the skin of theAnthrax lesion on the skin of the
forearm caused byforearm caused by Bacillus anthracisBa...
PATHOGENESIS
• Capsule > Invasiveness
– D-glutamic acid
• Exotoxin (Plasmid mediated)
i. Protective Factor (Antigen).
ii. ...
LABORATORY DIAGNOSIS:
• Specimens obtained from:
a malignant pustule, sputum, blood.
- Gram stain + fluorescent-antibody s...
Bicarbonate agar and blood agarBicarbonate agar and blood agar
plate cultures ofplate cultures of Bacillus anthracisBacill...
Negative encapsulation: Blood agar andNegative encapsulation: Blood agar and
bicarbonate agar plate cultures ofbicarbonate...
• TREATMENT
– Penicillin, Ciprofloxacin
• IMMUNIZATION
–Animals > Live spore vaccine
(Sterne strain)
–Workers at Risk of E...
ii.ii. Bacillus cereusBacillus cereus
• Food Poisoning.
• Clinical Syndromes:
i. Severe Nausea &Vomiting.
ii. Abdominal Cr...
PATHOGENICITY:
>> Due to an Enterotoxin.
• Also Causes Disease in Patients with
Underlying Disease.
• TREATMENT:
>> Tetrac...
2. GENUS:2. GENUS: CLOSTRIDIUMCLOSTRIDIUM
• Gram +ve bacilli
• Anaerobic,
• Spore Forming
- Spores:
Ink Stain of SporulatingInk Stain of Sporulating ClostridiumClostridium--
spores appear clear, vegetative cellsspores appe...
i.i. Clostridium perfringensClostridium perfringens
• Nonmotile
• Spores Not Produced in Ordinary
Media.
• Aerotolerant An...
Gram stain ofGram stain of Clostridium perfringensClostridium perfringens
Exudate smear ofExudate smear of
Clostridium perfringensClostridium perfringens
Tissue smear ofTissue smear of
Clostridium perfringensClostridium perfringens
DISEASE:
• Clostridial Myonecrosis.
• Less Severe Wound Infections.
• Food Poisoning.
Patient with gas gangrenePatient with gas gangrene
LABORATORY IDENTIFICATION
• In Chopped Meat - Glucose Medium:
• On BA:
• On Egg Yolk Agar:
>> Precipitation (Opalescence)....
Blood agar plate withBlood agar plate with Cl. perfringensCl. perfringens
characteristic double zone of hemolysischaracter...
PATHOGENICITY & CLINICAL INFECTION
 α-Toxin: Acts on Lecithin-Containing Lipo-
protein Complexes in the Cell Membrane.
• ...
FOOD POISONING:
• Cl. perfringens Type A >> Enterotoxin.
> Acute Abdominal Pain and Diarrhoea.
LABORATORY DIAGNOSIS:
• Important: Diagnosis of Clostridium
Myonecrosis Should Be Rapid and Made on
Clinical Grounds.
i. D...
TREATMENT:
• Clostridium Myonecrosis:
i. Surgical Removal of All Infected and
Necrotic Tissue.
ii. Antibiotic and Antitoxi...
Clostridia That May Be Associated
with Gas Gangrene:
• Cl. perfringens Type A
• Cl. septicum
• Cl. novyi Type A
• Cl. hist...
Human case of malignant edemaHuman case of malignant edema
caused bycaused by Cl. septicumCl. septicum
ii.ii. Clostridium tetaniClostridium tetani
• > Tetanus.
• > Terminal Spores with Drumstick
Appearance.
• > Obligate Anaer...
Clostridium tetaniClostridium tetani
Gram Positive RodsGram Positive Rods
Clostridium tetaniClostridium tetani
VIRULENCE FACTORS:
• Tetanus Toxin (Tetanospasmin) >
Neurotoxin.
i. An Intercellular Toxin Released by
Cellular Autolysis....
CLINICAL INFECTION & PATHOGENESIS
• "Tetanus is Generalized in Nature".
i. Unimmunized Rural Population.
ii. In Practice: ...
Drawing of a Soldier dying ofDrawing of a Soldier dying of
Tetanus (Opisthotonos)Tetanus (Opisthotonos)
A patient presented with facial tetany.A patient presented with facial tetany.
Note the contraction of the masseter andNot...
LABORATORY DIAGNOSIS:
• > Diagnosis on Clinical Grounds.
TREATMENT:
• i. Antitoxin.
• ii. Debridement of Wound and Removal...
PREVENTION:
> Prompt and Adequate Cleaning of
Wounds.
i. Active Immunity.
ii. Passive Immunity.
iii.iii. Clostridium botulinumClostridium botulinum
• > Botulism.
• > Gram +ve, Spore Forming Bacilli.
• > Strict Anaerobe.
Gram Stain ofGram Stain of Cl. botulinumCl. botulinum,,
Characteristic Long RodsCharacteristic Long Rods
A photomicrograph ofA photomicrograph of
Clostridium botulinumClostridium botulinum type Atype A
Blood Agar Plate withBlood Agar Plate with C. botulinumC. botulinum
VIRULENCE FACTORS
• Botulinum Toxin >>> Neurotoxin.
–Serologically 8 Toxins >>
A, B, C1, C2, D, E, F & G.
> Affect the Cho...
DISEASE IN HUMANS
1. Food - Borne Botulism:
> Incubation Period: 12-36 Hours to 8 days.
2. Infant Botulism:
LABORATORY DIA...
TREATMENT & PREVENTION
Important: Specific Treatment Should
Begin as Quick as Possible.
>Polyvalent Antitoxin >>> Immediat...
iv.iv. Clostridium difficileClostridium difficile
• Antibiotic Associated Colitis.
• Produce Two Major Protein Toxins
(A &...
Clostridium difficileClostridium difficile
Scanning electron micrograph ofScanning electron micrograph of
Clostridium difficleClostridium difficle
Intestinal Smear- Close AssociationIntestinal Smear- Close Association
ofof Cl. difficileCl. difficile with Neutrophilswit...
• Infection Can Be:
–Endogenous or Exogenous.
• Nosocomial Spread: Due to Spores.
LAB DIAGNOSIS:
1. Demonstration of Cytot...
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Bacillaceae-Lectures 8-11-Bacillus anthracis, B. cereus, Clostridium

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Bacillaceae-Lectures 8-11-Bacillus anthracis, B. cereus, Clostridium

  1. 1. ‫مممممم‬ ‫مممممم‬ ‫مممم‬ ‫ممم‬‫مممممم‬ ‫مممممم‬ ‫مممم‬ ‫ممم‬ FAMILY: BACILLACEAEFAMILY: BACILLACEAE Prof. Khalifa Sifaw GhengheshProf. Khalifa Sifaw Ghenghesh
  2. 2. 1. GENUS:1. GENUS: BACILLUSBACILLUS • Gram +ve bacilli • Aerobic • Spore-Forming
  3. 3. 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"
  4. 4. Bacillus anthracisBacillus anthracis
  5. 5. An electron micrograph of spores fromAn electron micrograph of spores from the Sterne strain ofthe Sterne strain of Bacillus anthracisBacillus anthracis
  6. 6. Bacillus anthracisBacillus anthracis McFaydean capsuleMcFaydean capsule stain, grown at 35stain, grown at 35oo C, in defibrinatedC, in defibrinated horse blood.horse blood.
  7. 7. DISEASE: • In Animals: >> Septicaemia. • In Humans: i. Cutaneous Anthrax > Malignant pustule ii. Pulmonary Anthrax (Wool-Sorter'sDisease). iii. Gastrointestinal Anthrax.
  8. 8. Cutaneous AnthraxCutaneous Anthrax
  9. 9. Anthrax lesion on the skin of theAnthrax lesion on the skin of the forearm caused byforearm caused by Bacillus anthracisBacillus anthracis
  10. 10. 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
  11. 11. 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:
  12. 12. Bicarbonate agar and blood agarBicarbonate agar and blood agar plate cultures ofplate cultures of Bacillus anthracisBacillus anthracis
  13. 13. Negative encapsulation: Blood agar andNegative encapsulation: Blood agar and bicarbonate agar plate cultures ofbicarbonate agar plate cultures of Bacillus cereusBacillus cereus
  14. 14. • TREATMENT – Penicillin, Ciprofloxacin • IMMUNIZATION –Animals > Live spore vaccine (Sterne strain) –Workers at Risk of Exposure > Anthrax Vaccine Absorbed (AVA) >> “Alum precipitated toxoid”
  15. 15. ii.ii. Bacillus cereusBacillus cereus • Food Poisoning. • Clinical Syndromes: i. Severe Nausea &Vomiting. ii. Abdominal Cramps & Diarrhoea.
  16. 16. PATHOGENICITY: >> Due to an Enterotoxin. • Also Causes Disease in Patients with Underlying Disease. • TREATMENT: >> Tetracycline, Erythromycin. • iii. B. subtilis: • iv. B. stearothermophilus.
  17. 17. 2. GENUS:2. GENUS: CLOSTRIDIUMCLOSTRIDIUM • Gram +ve bacilli • Anaerobic, • Spore Forming - Spores:
  18. 18. Ink Stain of SporulatingInk Stain of Sporulating ClostridiumClostridium-- spores appear clear, vegetative cellsspores appear clear, vegetative cells darkdark
  19. 19. i.i. Clostridium perfringensClostridium perfringens • Nonmotile • Spores Not Produced in Ordinary Media. • Aerotolerant Anaerobe. • 5 Types: A - E
  20. 20. Gram stain ofGram stain of Clostridium perfringensClostridium perfringens
  21. 21. Exudate smear ofExudate smear of Clostridium perfringensClostridium perfringens
  22. 22. Tissue smear ofTissue smear of Clostridium perfringensClostridium perfringens
  23. 23. DISEASE: • Clostridial Myonecrosis. • Less Severe Wound Infections. • Food Poisoning.
  24. 24. Patient with gas gangrenePatient with gas gangrene
  25. 25. LABORATORY IDENTIFICATION • In Chopped Meat - Glucose Medium: • On BA: • On Egg Yolk Agar: >> Precipitation (Opalescence). • Milk Media: Stormy Formation. • Nagler Reacrion:
  26. 26. Blood agar plate withBlood agar plate with Cl. perfringensCl. perfringens characteristic double zone of hemolysischaracteristic double zone of hemolysis
  27. 27. PATHOGENICITY & CLINICAL INFECTION  α-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.
  28. 28. FOOD POISONING: • Cl. perfringens Type A >> Enterotoxin. > Acute Abdominal Pain and Diarrhoea.
  29. 29. 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.
  30. 30. TREATMENT: • Clostridium Myonecrosis: i. Surgical Removal of All Infected and Necrotic Tissue. ii. Antibiotic and Antitoxin Therapy. iii. Adminstration of Hyperbaric Oxygen. • Food Poisoning:
  31. 31. Clostridia That May Be Associated with Gas Gangrene: • Cl. perfringens Type A • Cl. septicum • Cl. novyi Type A • Cl. histolyticum • Cl. Sordellii
  32. 32. Human case of malignant edemaHuman case of malignant edema caused bycaused by Cl. septicumCl. septicum
  33. 33. ii.ii. Clostridium tetaniClostridium tetani • > Tetanus. • > Terminal Spores with Drumstick Appearance. • > Obligate Anaerobe.
  34. 34. Clostridium tetaniClostridium tetani Gram Positive RodsGram Positive Rods
  35. 35. Clostridium tetaniClostridium tetani
  36. 36. VIRULENCE FACTORS: • Tetanus Toxin (Tetanospasmin) > Neurotoxin. i. An Intercellular Toxin Released by Cellular Autolysis. ii. Inhibits the Release of Inhibitory Transmitters. iii. Toxoid.
  37. 37. 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:
  38. 38. Drawing of a Soldier dying ofDrawing of a Soldier dying of Tetanus (Opisthotonos)Tetanus (Opisthotonos)
  39. 39. A patient presented with facial tetany.A patient presented with facial tetany. Note the contraction of the masseter andNote the contraction of the masseter and neck musclesneck muscles
  40. 40. 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.
  41. 41. PREVENTION: > Prompt and Adequate Cleaning of Wounds. i. Active Immunity. ii. Passive Immunity.
  42. 42. iii.iii. Clostridium botulinumClostridium botulinum • > Botulism. • > Gram +ve, Spore Forming Bacilli. • > Strict Anaerobe.
  43. 43. Gram Stain ofGram Stain of Cl. botulinumCl. botulinum,, Characteristic Long RodsCharacteristic Long Rods
  44. 44. A photomicrograph ofA photomicrograph of Clostridium botulinumClostridium botulinum type Atype A
  45. 45. Blood Agar Plate withBlood Agar Plate with C. botulinumC. botulinum
  46. 46. 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).
  47. 47. DISEASE IN HUMANS 1. Food - Borne Botulism: > Incubation Period: 12-36 Hours to 8 days. 2. Infant Botulism: LABORATORY DIAGNOSIS i. Diagnosis Made Clinically. ii. Detection of Organism or Its Toxin in the Suspected Food iii. Samples of Stool or Vomit
  48. 48. TREATMENT & PREVENTION Important: Specific Treatment Should Begin as Quick as Possible. >Polyvalent Antitoxin >>> Immediately. >Physiological Support >>> ICU. >NEVER Use a Swollen or Defective Can.
  49. 49. iv.iv. Clostridium difficileClostridium difficile • Antibiotic Associated Colitis. • Produce Two Major Protein Toxins (A &B). • Risk Factors: –Antibiotic Exposure. –Old Age.
  50. 50. Clostridium difficileClostridium difficile
  51. 51. Scanning electron micrograph ofScanning electron micrograph of Clostridium difficleClostridium difficle
  52. 52. Intestinal Smear- Close AssociationIntestinal Smear- Close Association ofof Cl. difficileCl. difficile with Neutrophilswith Neutrophils
  53. 53. • 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.

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