pseudomonas 02.04.16
TRANSCRIPT
Dr.Namrata
Vibrios requiring a higher concentration of sodium chloride
natural inhabitants of sea water and marine life
V. parahaemolyticus, Vibrio alginolyticus,V. vulnificus - important halophilic vibriosspecies known to cause infection in humans
important cause of seafood-associated gas
troenteritis throughout the world
differs from V. cholerae by having a capsule showing bipolar staining and pleomorphism
it has polar flagella when grown in liquid culture, but shows peritrichous flagella whengrown on solid media.
salt concentration : up to 8%
Growth characters:
Mac:nonlactose- fermenting colonies
Blood agar:beta-hemolytic colonies
TCBS:nonsucrose-fermenting green colonies
oxidase – positive
Catalase - positive
Antigens:
somatic O antigens,
flagellar H antigens,
capsular K antigen.
Serotyping is based on O and K antigen.
Virulence factors:
Thermostable direct hemolysin is the key virulence factor of V. parahaemolyticus.
This is found only in strains that are pathogenic to human beings.
Kanagawa phenomenon
Pathogenic strains of V. parahaemolyticuswhen grown on a special high-salt mannitol medium (Wagatsuma agar) show hemolysison blood agar
Nonpathogenic strains isolated from environment that do not cause any hemolysis
Habitat:
is widely distributed in estuary and marine environments.
Seafood, such as fish, crabs, or oysters, is the main source and reservoir of infection.
The infection is acquired on consumption of contaminated seafood.
Clinical manifestations:
in humans causes gastroenteritis
severity of the condition can vary from mild self-limited diarrhea to an acute illness
nausea,vomiting, abdominal pain, and low-grade fever, which may be present for 3 days
Stool usually does not contain any blood or mucus, but contains cellular exudates
Extraintestinal infections:
wound infections and ear and eye infections
in individuals exposed to contaminated sea water
V. alginolyticus is widely distributed in sea water and seafood.
high salt tolerance and can grow even in the presence of 10% sodium chloride
TCBS agar: large yellow sucrose-fermenting colonies.
shows swarming on the surface of nonselective medium, such as blood agar
associated with infections of superficial wounds exposed to contaminated sea water
has been associated with infections of the
gastrointestinal tract, ear, and eye.
Properties Vibrioparahaemolyticus
Vibrioalginolyticus
Growth in 10% agar - +
Presence of swarming
- +
Fermentation of sucrose
- +
VP test - +
formerly known as L vibrio,or Benekeavulnificus
TCBS: produces green nonsucrose-fermenting
colonies on TCBS medium
differs from V. parahaemolyticus and other Vibrio species by its ability to ferment lactose
Virulence:
more virulent bacterium than V.parahaemolyticus.
capsule
hydrolytic enzymes, such as cytolysins, proteases, and collagenases
resistant to complement- and antibody-mediated lysis
Clinical manifestations:
wound infection and
gastroenteritis rapidly progressing to septicemia
Wound infections:
caused by contaminated sea water
short incubation period of 3–24 hours
wounds are usually present on the fingers, palms, or soles of the feet
rapidly progresses to necrosis, gangrene,ornecrotizing fasciitis
Gastroenteritis:
following the consumption of raw seafood on exposure
rapidly progresses to septicaemia
manifests as multiple hemorrhagic bullae and extensive ecchymosis distributed on the lower extremities
Hypotension , oliguria and noncardiogenicpulmonary edema
V. vulnificus produces life-threatening illness,
immediate and prompt treatment with antibiotics is essential.
Tetracyclines or aminoglycosides are the antibiotics of choice
The condition is prevented by avoidance of raw and undercooked seafood.
PSEUDOMONAS
A large group of aerobic, non sporing
gram negative bacteria motile by polar
flagella
Found in water, soil, other moist
environments
Some of them are pathogenic to plants
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Classification
◦ Based on rRNA – DNA homology groups
Five ribosomal RNA groups
◦ Based on Phenotypic characters
Seven groups – Fluroscent, stutzeri, alcaligens,
pseudomallei, acidovorans, facilis & diminuta
Gram negative bacteria
Size 1.5 – 3 x 0.5 µ m
Actively motile by polar flagella
Produces extracellular polysaccharide k/a
alginate
Obligate aerobe
Grows at a temp range of 6 - 420 C
Produces colonies with distinctive musty, mawkish or earthy smell.
Growth on Blood agar -
colonies are moist flat, irregular edges,
greenish pigmented, showing zone of
hemolysis
Growth on MacConkey agar –
Non lactose fermenting colonies
Growth on Nutrient agar –
Greenish pigment
Pyocin
Pyorubin
Pyomelanin
Fluroscein
Selective media
Cetrimide agar
Pigment enhancing media
King’s A
King’s B
Cell surface
◦ Alginate
◦ Lipopolysaccharide
◦ Pili
◦ Flagella
Outer membrane
◦ Siderophore receptors
◦ Efflux pumps
Type III secretions
Secreted proteins
◦ Las A
◦ Las B
Iron acquisition
◦ Pyoverdine
◦ Pyochelin
Secreted toxins
◦ Exotoxin A
◦ Leucocidin
◦ Phospholipidases
Quorum sensing
Complex and diverse
Rarely causes infection in healthy individuals
Disruption of cutaneous / mucosal barrier
◦ Burns
◦ Dermatitis
◦ Penetrating trauma
◦ Surgery
◦ Endotracheal intubation
◦ Catheterisation
Immunosuppression
◦ Neutropenia
◦ Hypogamaglobulinemia
◦ Defective CMI
◦ Extremes of age
◦ Diabetes mellitus
◦ Steroids
◦ Cystic fibrosis
◦ AIDS
◦ cancer
Disturbed normal flora
◦ Long term hospital stay
◦ Broad spectrum antibiotics
Host factorsBacterial factors
Attachment
Invasion
Epithilium
Soluble factors
Anaphylotoxins
Neutrophils
Complement
factors
Pili
Flagella
Alginate
LPS
OMP
Type III secretion
LPS
Elastase
Proteases
SiderophoresToxins
Can cause infection anywhere
In adults mostly associated as a nosocomial
pathogen
In paediatric population it mainly causes
infection in cystic fibrosis patients
Superficial infections◦ Burn wound infections
◦ Otitis externa / Malignant otitis externa
◦ Pseudomonas keratitis
Systemic infections
◦ Respiratory tract infections
◦ Bacterimia
◦ Endocarditis
◦ CNS infections
◦ Urinary tract infections
◦ Bone and joint infections
Hospital acquired infections
Pneumonia
◦ Endotracheal tubes
◦ Ventilator tubings
◦ Humidifiers
◦ Nebulizers
Catheter associated infections
◦ UTI
◦ Line associated bacterimias
Iatarogenic meningitis
◦ Post lumbar puncture
◦ Post neurosurgery
Sample collection
◦ Respiratory tract infection
Sputum
Bronchioalveolar lavage
Bronchial washings
◦ CNS infections
CSF
◦ Bacteremia
Blood
◦ Eye infections
Corneal scrapings
Contact lenses
Lens fluid
◦ Ear infection
Ear swabs
◦ Superficial infections
Wound swabs
Microscopy Gram’s staining
Culture
◦ BA
◦ MA
◦ Selective medium
Biochemical reactions
◦ Oxidase
◦ Hugh & Leifson’s OF reactions
Antibiotic sensitivity
Pyocin typing
Molecular methods
Aminoglycosides
Gentamycin, Amikacin, Cephalosporins
Cefotaxime. Ceftazidime. Ofloxacin,
Piperacillin, ticarcillin
Local application, colistin, polymyxin
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Septicaemia Endocarditis Ecthyma
gangrenous Infantile diarrhoea Shanghai fever Disabling eye
infections Survive with
minimal nutrients
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Patients with cystic fibrosis are highly susceptible to pseudomonas lung infections.
cancer
burn patients
immunocompromised..
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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
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Respirators Endotracheal tubes
Can be Infected
All equipment's to be sterilized
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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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Diagnosis of P,aeroginosa infection depends upon isolation and laboratory identification of the bacterium.
blood agar / eosin-methylthionine blue agar
Grams
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.s aeruginosa colonies. Fluorescence is also used to suggest the presence of P. aeruginosa in wounds.
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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.
Thank you