‫بسم اهلل الرحمن الرحيم‬

FAMILY: BACILLIACEAE
Prof. Khalifa Sifaw Ghenghesh
 1. GENUS: BACILLUS

• Gram +ve bacilli

• Aerobic
• Spore-Forming
 i. Bacillus 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"
Bacillus anthracis
An electron micrograph of spores from
the Sterne strain of Bacillus anthracis
Bacillus anthracis McFaydean capsule
stain, grown at 35oC, in defibrinated
horse blood.
DISEASE:
• In Animals: >> Septicaemia.

• In Humans:
i. Cutaneous Anthrax > Malignant pustule
ii. Pulmonary Anthrax (Wool-Sorter'sDisease).
iii. Gastrointestinal Anthrax.
Cutaneous Anthrax
 Anthrax lesion on the skin of the
forearm caused by Bacillus anthracis
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
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:
 Bicarbonate agar and blood agar
plate cultures of Bacillus anthracis
Negative encapsulation: Blood agar and
bicarbonate agar plate cultures of
Bacillus cereus
• TREATMENT
– Penicillin, Ciprofloxacin

• IMMUNIZATION
– Animals > Live spore vaccine
(Sterne strain)
– Workers at Risk of Exposure >
Anthrax Vaccine Absorbed (AVA) >>
“Alum precipitated toxoid”
 ii. Bacillus cereus

• Food Poisoning.

• Clinical Syndromes:
i. Severe Nausea &Vomiting.
ii. Abdominal Cramps & Diarrhoea.
PATHOGENICITY:
>> Due to an Enterotoxin.
• Also Causes Disease in Patients with
Underlying Disease.

• TREATMENT:
>> Tetracycline, Erythromycin.

• iii. B. subtilis:
• iv. B. stearothermophilus.
 2. GENUS: CLOSTRIDIUM

• Gram +ve bacilli

• Anaerobic,

• Spore Forming
- Spores:
 Ink Stain of Sporulating Clostridium-
spores appear clear, vegetative cells dark
 i. Clostridium perfringens

• Nonmotile

• Spores Not Produced in Ordinary

Media.

• Aerotolerant Anaerobe.

• 5 Types: A - E
Gram stain of Clostridium perfringens
 Exudate smear of
Clostridium perfringens
 Tissue smear of
Clostridium perfringens
DISEASE:

• Clostridial Myonecrosis.

• Less Severe Wound Infections.

• Food Poisoning.
Patient with gas gangrene
LABORATORY IDENTIFICATION
• In Chopped Meat - Glucose Medium:
• On BA:
• On Egg Yolk Agar:
>> Precipitation (Opalescence).

• Milk Media: Stormy Formation.

• Nagler Reacrion:
 Blood agar plate with Cl. perfringens
characteristic double zone of hemolysis
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.
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. Direct Smear and Gram Stain of Material

from Deep Within the Wound.

ii. Culture:
Tissue Aspirates or Deep Swabs Taken
from Affected Muscle.
TREATMENT:
• Clostridium Myonecrosis:
i. Surgical Removal of All Infected and
Necrotic Tissue.
ii. Antibiotic and Antitoxin Therapy.
iii. Adminstration of Hyperbaric Oxygen.

• Food Poisoning:
 Clostridia That May Be Associated
with Gas Gangrene:

• Cl. perfringens Type A

• Cl. septicum
• Cl. novyi Type A
• Cl. histolyticum
• Cl. Sordellii
Human case of malignant edema
caused by Cl. septicum
 ii. Clostridium tetani

• > Tetanus.
• > Terminal Spores with Drumstick
Appearance.
• > Obligate Anaerobe.
Clostridium tetani
Gram Positive Rods
Clostridium tetani
VIRULENCE FACTORS:
• Tetanus Toxin (Tetanospasmin) >
Neurotoxin.
i. An Intercellular Toxin Released by
Cellular Autolysis.
ii. Inhibits the Release of Inhibitory
Transmitters.
iii. Toxoid.
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:
Drawing of a Soldier dying of
Tetanus (Opisthotonos)
 A patient presented with facial tetany.
Note the contraction of the masseter and
neck muscles
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.
PREVENTION:
> Prompt and Adequate Cleaning of
Wounds.

i. Active Immunity.
ii. Passive Immunity.
 iii. Clostridium botulinum

• > Botulism.
• > Gram +ve, Spore Forming Bacilli.
• > Strict Anaerobe.
Gram Stain of Cl. botulinum,
Characteristic Long Rods
 A photomicrograph of
Clostridium botulinum type A
Blood Agar Plate with C. botulinum
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).
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
TREATMENT & PREVENTION
Important: Specific Treatment Should
Begin as Quick as Possible.

>Polyvalent Antitoxin >>> Immediately.

>Physiological Support >>> ICU.
>NEVER Use a Swollen or Defective Can.
 iv. Clostridium difficile

• Antibiotic Associated Colitis.

• Produce Two Major Protein Toxins
(A &B).
• Risk Factors:
– Antibiotic Exposure.
– Old Age.
Clostridium difficile
Scanning electron micrograph of
Clostridium difficle
Intestinal Smear- Close Association
of Cl. difficile with Neutrophils
• 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.