PSEUDOMONAS

Dr.T.V.Rao MD

DR.T.V.RAO MD

PSEUDOMONAS
A large group of aerobic, non sporing gram negative bacteria motile by polar flagella Found I nature water, soil, other moist environments Some of them are pathogenic to plants

Creation of new genera such as Burkholderia. Stenotrophomnonas

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GENERAL CHARACTERISTICS
- Widely distributed in soil and water - Gram negative rods - Aerobic - Motile - Produce water-soluble pigments Opportunistic pathogens
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MORPHOLOGY
They are slender gram negative bacillus, 1.5 3 microbes x 0.5 microns Monoflgellar ? Non capsulated but many strains have mucoid slime layer

Isolates from Cystic fibrosis patients have abundance of extracellular polysaccharides composed of alginate polymers
Escape the defence mechanisms by loose capsule in which micro colonies of bacillus are enmeshed and protected from host defences.

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P. aeruginosa
Forms round colonies with a fluorescent greenish color, sweet odor, and b-hemolysis. Pyocyanin- nonfluorescent bluish pigment; pyoverdin- fluorescent greenish pigment; pyorubin, and pyomelanin Some strains have a prominent capsule (alginate). Identification of P. aeruginosa is usually based on oxidase test and its colonial morphology: b-hemolysis, the presence of characteristic pigments and sweet odor, and growth at 42 oC.

CULTURAL CHARACTERS
Obligate aerobe, but grow anaerobically if nitrate is available Growth occurs at wide range of temperatures 6-42 c the optimum being 37 c Growth on ordinary media producing large opaque irregular colonies with distinctive musty mawkish or earthy smell.

Iridescent patches with metallic sheen are seen in cultures on nutrient agar.
In broth forms dense turbidity with surface pellicle.
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Characteristics of Pseudomonas aeruginosa Motile (by single or multiple polar flagella) gram-negative rods Obligate (strict) aerobes (most strains) Oxidase (usually) and catalase positive Nonfermentative chemoheterotrophic respiratory metabolism Minimal nutritional reqts.; Many organic compounds used as C and N sources, but only a few carbohydrates by oxidative metabolism Glucose used oxidatively Lactose negative on MacConkeys agar
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PIGMENT PRODUCTION
Some strains produce diffusible pigments: Pyocyanin (blue); fluorescein (yellow); pyorubin (red)

P. aeruginosa produces characteristic grape-like odor and blue-green pus & colonies
Broad antibiotic resistance
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BIOCHEMICAL REACTIONS
Oxidative and Non fermentative Glucose is utilized oxidatively Indole, MR and VP and H2 S tests are negative Catalase, Oxidase, and Arginine tests are positive
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TYPING AND IMPORTANCE

Important cause of Hospital Infections Important for epidemiological purpose Serotyping

Bacteriocins typing
Pyocyanin Aeruginosin typing Restriction endonuclease typing with pulsed gel electrophoresis
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RESISTANCE
Killed at 55oc in on 1 hour High resistance to chemical agents Resistance to quaternary ammonium compounds.Chlorxylenol Resistant to Hexchlorophenes Grows also in antiseptic bottles

Dettol as cetrimide as selective medium

Sensitive to acids silver salts, beta glutaraldehyde
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WHAT ANTIBIOTICS TO USE

Aminoglycosides Gentamycin, Amikacin, Cephalosporins Cefotaxime. Ceftazidime. Ofloxacin, Piperacillin, ticarcillin Local application, colistin, polymyxin

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PATHOGENICITY
Blue pus Causing the nosocomial infection Suppurative otitis

Localised and generalised infections

Urinary tract infection after catheterization Iatrogenic meningitis Post tracheostomy pulmonary infections
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Pathogenesis and Immunity

P. aeruginosa

This organism is widely distributed in nature and is commonly present in moist environments in hospitals. It is pathogenic only when introduced into areas devoid of normal defenses, e.g., 1. Disruption of mucous membrane and skin. 2. Usage of intravenous or urinary catheters. 3. Neutropenia (as in cancer therapy).

P. aeruginosa
Pathogenesis

Antigenic structure, enzymes, and toxins

Pili and nonpilus adhesins. Capsule (alginate, glycocalyx): seen in cultures from patients with cystic fibrosis. LPS- endotoxin, multiple immunotypes. Pyocyanin: catalyzes production of toxic forms of oxygen that cause tissue damage. It also induces IL-8 production. Pyoverdin: a siderophore.

Proteases
Serine protease, metalloprotease and alkaline protease cause tissue damage and help bacteria spread. Phospholipase C: a hemolysin Exotoxin A: causes tissue necrosis and is lethal for animals (disrupts protein synthesis); immunosuppressive.

Exoenzyme S and T: cytotoxic to host cells.

PATHOGENESIS AND IMMUNITY

P. aeruginosa can infect almost any external site or organ.
P. aeruginosa is invasive and toxigenic. It attaches to and colonizes the mucous membrane or skin, invade locally, and produces systemic diseases and septicemia.
P. aeruginosa is resistant to many antibiotics. It becomes dominant when more susceptible bacteria of the normal flora are suppressed.

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CLINICAL PRESENTATIONS
Septicaemia

Endocarditis
Ecthyma gangrenous Infantile diarrhoea Shanghai fever Disabling eye infections Survive with minimal nutrients

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WHO IS MORE SUSCEPTIBLE TO INFECTION

This bacterium is of particular concern to individuals with cystic fibrosis who are highly susceptible to pseudomonas lung infections. Pseudomonas aeruginosa is also of grave concern to cancer and burn patients as well as those people who are immunocompromised. The case fatality rate for individuals infected with Pseudomonas aeruginosa approaches 50 percent.
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PSEUDOMONAS AND CYSTIC FIBROSIS

Pseudomonas aeruginosa is the most frequently encountered lung pathogen in patients with cystic fibrosis (CF). Following initial, often intermittent, episodes of infection, it becomes a permanently established component of the chronically infected lung in more than 80% of patients and confers an adverse prognosis
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INFECTION OF EQUIPMENT'S

Respirators Endotracheal tubes Can be Infected All equipment's to be sterilized

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PSEUDOMONAS AND URINARY TRACT INFECTIONS

Pseudomonal UTIs are usually hospital-acquired and are associated with catheterization, instrumentation, and surgery. These infections can involve the urinary tract through an ascending infection or through bacteriuic spread. In addition, these infections are a frequent source of bacteraemia. No specific characteristics distinguish this type of infection from other forms of UTI.
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TOXINS AND ENZYMES IN PSEUDOMONAS

Toxic extracellular products in culture filtrates Exotoxin A and S Exotoxin A acts as NADase resembling Diphtheria toxin Proteases,elastatese hemolysins and enterotoxin Slime layer and Biofilms

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PSEUDOMONAS AERUGINOSA AN IMPORTANT OPPORTUNISTIC PATHOGEN

Pseudomonas aeruginosa is an opportunistic pathogen, meaning that it exploits some break in the host defences to initiate an infection. In fact, Pseudomonas aeruginosa is the epitome of an opportunistic pathogen of humans. The bacterium almost never infects uncompromised tissues, yet there is hardly any tissue that it cannot infect if the tissue defences are compromised in some manner
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P.AEROGINOSA IS AN OPPORTUNISTIC PATHOGEN

P,aeroginosa is an opportunistic pathogen. It rarely causes disease in healthy persons. In most cases of infection, the integrity of a physical barrier to infection (eg, skin, mucous membrane) is lost or an underlying immune deficiency (eg, neutropenia, immunosuppression) is present. Adding to its pathogenicity, this bacterium has minimal nutritional requirements and can tolerate a wide variety of physical conditions
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PSEUDOMONAS PROMINENT HOSPITAL ACQUIRED INFECTIONS

It causes urinary tract infections, respiratory system infections, dermatitis, soft tissue infections, bacteraemia, bone and joint infections, gastrointestinal infections and a variety of systemic infections, particularly in patients with severe burns and in cancer and AIDS patients who are immunosuppressed.
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DIAGNOSIS OF P.AEROGINOSA INFECTION

Diagnosis of P,aeroginosa infection depends upon isolation and laboratory identification of the bacterium. It grows well on most laboratory media and commonly is isolated on blood agar or eosin-methylthionine blue agar. It is identified on the basis of its Gram morphology, inability to ferment lactose, a positive oxidase reaction, its fruity odour, and its ability to grow at 42C. Fluorescence under ultraviolet light is helpful in early identification of P.s aeruginosa colonies. Fluorescence is also used to suggest the presence of P. aeruginosa in wounds.
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IDENTIFICATION WITH CHROMAGAR

Pseudomonas sp. develop as easily distinguishable blue-green coloured colonies, clearly visible under normal lighting conditions. Other bacterial species are inhibited or give colourless colonies. Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas putida and Pseudomonas fragilis all give typical blue-green colony colouration and can be studied directly by serotyping or biochemical methods.

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LABORATORY IDENTIFICATION OF DIAGNOSIS OF P.AEROGINOSA INFECTIONS

Diagnosis of P. aeruginosa infection depends upon isolation and laboratory identification of the bacterium. It grows well on most laboratory media and commonly is isolated on blood agar or eosin-methylthionine blue agar. It is identified on the basis of its Gram morphology, inability to ferment lactose, a positive oxidase reaction, its fruity odour, and its ability to grow at 42 C. Fluorescence under ultraviolet light is helpful in early identification of P. aeruginosa colonies and may also help identify its presence in wounds.
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TREATING PSEUDOMONAS INFECTIONS Combined antibiotic therapy is generally required to avoid resistance that develops rapidly when single drugs are employed. Avoid using inappropriate broad-spectrum antibiotics, which can suppress the normal flora and permit overgrowth of resistant pseudomonads.
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PSEUDOMONAS AERUGINOSA A RESISTANT PATHOGEN

Pseudomonas aeruginosa is frequently resistant to many commonly used antibiotics. Although many strains are susceptible to gentamicin, tobramycin, colistin, and amikacin, resistant forms have developed. The combination of gentamicin and carbenicillin is frequently used to treat severe Pseudomonas infections. Several types of vaccines are being tested, but none is currently available for general use.
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P. aeruginosa
Prevention and Control Pseudomonas spp. normally inhabit soil, water, and vegetation and can be isolated from the skin, throat, and stool of healthy persons. Spread is mainly via contaminated sterile equipment's and cross-contamination of patients by medical personnel. High risk population: patients receiving broad-spectrum antibiotics, with leukemia, burns, cystic fibrosis, and

immunosuppression.
Methods for control of infection are similar to those for other nosocomial pathogens. Special attention should be paid to sinks, water baths, showers, hot tubs, and other wet areas.

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