FAMILY: BACILLACEAE

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 Clostridiumspores 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 Lipoprotein 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.