BACILLUS

BACILLUS ANTHRACIS

Gram posiotive bacilli nonbranching aerobic corynebacterium anaerobic clostridia aerobic branching anaerobic

bacillus

listeria

gardenerella

lactobacillus

Spore forming gram positive bacilli Aerobic anthrax anthracoid Anaerobic clostridia

BACILLUS GROUP

They are aerobes, gram positive and spore forming rods. The most important pathogenic member in this genus is Bacillus anthracis Most of the other members are saprophytes in water, soil and air. They are collectively termed anthracoid. Of these, B. cereus and B subtilis may act as opportunistic pathogens in debilitated or immunocompromised persons.

Bacillus anthracis

Morphology:

Gram positive rectangular large organisms arranged in chains.

polychrome methylene blue the capsule appears as pink rim around

They are capsulated in vivo;

when stained with

the blue bacillus

They form spores in vitro. The spores are central, ovoid and are not stained with gram.

Bacillus anthracis Gram stain. 1500X The cells have characteristic squared ends. The endospores are ellipsoidal shaped and located centrally in the sporangium. The spores are highly refractile to light and resistant to staining

Culture characters:

Aerobes grow on nutrient agar at 37C A 24 colony on agar is large opaque disc with a rough granular surface and irregular fimbriate edge ( medusa head colony). It does not hemolyse blood.

Antigenic structure and virulence:

A poypeptide capsule made up exclusively of Dglutamic acid exhibits antiphagocytic activity but does not stimulate protective antibody. The exotoxin is a heat labile protein composed of three components referred to as

Protective antigen (PA) and lethal factor (LF) , which is responsible for most of the
toxicity, and

edema factor (EF). Together they are cytolytic for macrophages , tissues and increase vascular permeability causing edema and shock

Pathogenesis

Anthrax is primarily a disease of farm animal's e.g. cattle and sheep. Man is infected by coming in contact with infected animals or their dead bodies. Humans are infected by spores on animal products such as hides, bristles, and wool. Individuals more liable to get the infection are farmers, butchers and veterinarians.

Cutaneous anthrax (malignant pustule).

The organism enters through skin abrasions or cuts and germinates.

The vegetative cells multiply locally and the host responds with an acute inflammatory response characterized by an influx of polymorphonuclear leukocytes. Phagocytosis by polymorphonuclear leukocytes is inhibited by the capsule and the organisms continue to survive and multiply.

The organisms release exotoxin locally and begin to invade adjacent tissue rapidly producing extensive damage. Within 2-5 days, this process manifests as malignant pustule , painless necrotic ulcer with black crust &local edema. The lesion heals very slowly after the eschar separates. Less commonly, the disease progress with massive local edema, toxemia and bacteremia. It is a fatal outcome if untreated

Over the course of a few days, cutaneous anthrax

develops into a sore with a coal-black center

Pulmonary anthrax (wool sorters disease)

The disease is acquired by inhalation of vegetative cells in droplet nuclei from a patient with respiratory disease or by inhalation of spores from infected animals or animal products or following laboratory accident. When the spores are introduce they germinate into vegetative cells in the trachea , bronchi and/or lung.

Cont.

Vegetative cells undergo multiplaction in these organs with the host response, and exotoxin release as described for the cutaneous form of the disease. Within 1-5 days, pulmonary necrosis and respiratory distress, septicemia, and meningitis occur which is most often and rapidly fatal despite therapy. Today it is the form we would expect from the dissemination of a spore aerosol in biologic

warfare

Diagnosis:

Pathologic samples e.g. exudates from skin lesions, blood & CSF examined as follows:

Smears are stained with gram

Anthrax can be identified in dried smear by immunofluorescence staining techniques. This is a conclusive specific method of diagnosis. PCR
Cultures on nutrient agar. Colonies that arise are injected into mice or guinea pigs. Serologic test

Treatment:

Penicillin, tetracyclines, are effective in the treatment of cutaneous anthrax only when given early in the course of the disease. The effectiveness of antimicrobial therapy among patients with inhalation anthrax is minimal.

Prevention:

Disposal of animal carcases by burning or by deep burial in lime pits. Autoclaving of animal products. Protective clothing and gloves for handling potentially infected materials. Active immunization of domestic animals with live attenuated vaccines.

Persons with high occupational risk should be immunized with a cell free vaccine induces short lived immunity and requires annual boosters.

PSEUDOMONAS

Pseudomonas aeruginosa is the commonest human pathogen of the pseudomonas group. It is found in soil, sewage and water, some are commensal in the intestine; others cause urinary tract infection, wound infection and otitis media. The organism is highly resistant to antibiotics and causes chronic infections especially in debilitated, old and hospitalized patients with low defense mechanisms.

Morphology, Culture characters & Biochemical reaction

Morphology Gram negative motile bacilli. Culture characters: Aerobe grows on nutrient agar leading to greenish coloration of the medium due to its diffusible exopigment which consists of pyocyanin (blue green) and fluorescein (yellow). Biochemical reaction:

It is oxidase positive. It does no ferment any sugar. show the characteristic greenish colouration of the medium.

Colonies of Pseudomonas aeruginosa growing on an agar plate

Antigenic structure, toxin and enzyme

Pili promote attachment to host epithelial cells. Polysaccharide capsule which responsible for the mucoid colonies seen in culture from patients with cystic fibrosis. Extracellular enzyme including elastases, protease and hemolysin. Exotoxin A which causes tissue necrosis through blocking of protein synthesis. Lipopolysaccharide ( endotoxin).

Pathogeneses:

Skin infections e.g burn sites, wounds pressure sore, ulcer, etc. Urinary tract infection, usually following catheterization or associated with chronic urinary tract infection. Respiratory infection especially in patient with cystic fibrosis. External ear infection. Eye infection (hospital acquired).

Diagnosis of infections:
The pus from the lesions may be greenish in colour. Smears stained by gram show gram negative bacilli among pus cells. Cultures on nutrient agar

Treatment:

Is difficult due to resistance of the organism to cmommon antibiotics. Antibiotic sensitivity tests should be done to find the proper antibiotic. Aminoglycosides e.g. gentamycin or amikin used in combination with penicillin active against Ps. aeruginosa e.g. ticarcillin, mezlocillin or piperacillin are used.

Moraxella

Moraxella is the genus in the family Neisseriaceae

Moraxella catarrhalis

is a gram negative diplococcus. It is a member of the normal flora in human oropharynx, it can cause sinusitis, otitis , bronchitis and pneumonia, particularly in immunocompromised patients. Most clinical isolates produce lactamase.

Moraxella lacunata is a member in this genous that

causes angular conjunctivitis. They appear in smears from conjunctival discharge as gram negative diplococci

Klebsiella

Klebsiella are normal inhabitants of the intestine and respiratory tract.

They are sabrophytes in soil and water. Some are cause disease in man

Morphology: gram negative bacilli, non-motile and capsulated. Cultural characters: they give pink colonies on MacConkey. Colonies are mucoid due to the production of abundant extracellular slim. Biochemical activities:

They ferment glucose, lactose, maltose, mannite and sucrose with production of acid and gas. They are indol negative, M.R negative ,V.P positive and citrate positive.

Diseases

Kl.pneumoniae (Freeidlanders bacillus):

It causes lobar pneumoniae in man. It is highly pathogenic to mice and cause their death within 24hs, when injected intraperitonally. It causes urinary tract infection.

Kl. Rhinoscleromatis causes rhinoscleroma, which is a granulomatous lesion in the nose and throat

Thank you

The anthrax spore and its several protective layers Anthrax can be transmitted by three routes: Inhalation Cutaneous Gastrointestinal