Pseudomonads

Ni Nyoman Nami Arthisari

197119004
Pembimbing: Dr. dr. Ida Sri Iswari, Sp.MK, M.Kes
Talaro K.P., Chess B. 2012. Foundation in Microbiology. Ed 8. Pasadena. Mc Graw Hill.
Gram Negative Bateria (bacilli)
Pseudomonads
 Pseudomonads

 members of the genus Pseudomonas and several

Pseudomonas-like organisms, include:
● Gram-negative bacillus or coccobacillus
● Strictly aerobic metabolism
● Motile usually with polar flagellum or polar tuft of flagella
● Oxidase positive (except P. luteolus and P. oryzihabitans)
● Catalase positive
● Usually grows on MAC agar
● Usually an oxidizer of carbohydrates, but some species are
asaccharolytic
● The pseudomonads occur widely in soil, water, plants, and
animals.
 • The classification of pseudomonads is
based on rRNA/DNA homology and
common culture characteristics.
• P aeruginosa is frequently present in
small numbers in the normal intestinal
flora and on the skin of humans, and is
the major pathogen of the group. Other
pseudomonads infrequently cause
disease.
• Some produce biofilm

Cornelissen C.N., Fisher B.D., Harvey R.A. 2001. Lippincott’s Illustrated Reviews: Microbiology. Ed 3. Philadelphia. Lippincott.
Brooks G.L et al. Jawetz, Melnick, & Adelberg’s Medical Microbiology. Ed 27. New York. 2016. Lange.
Pseudomonad
group: habitat and
mode of
transmission

Tille P.M. 2017. Bailey & Scott’s Diagnostic

Microbiology: Bacterial Genetics, Metabolism, and
Structure. Ed 14. Missouri. Elsevier.
 Pseudomonas aeruginosa

● Small, gram-negative rods with a single polar flagellum

● Aerobic and does not ferment lactose (traits that distinguish it from
members of the Enterobacteriaceae).
● Oxidase positive.
● Most commonly isolated in clinical specimens
● It is found in moist environments, pools, hot tubs, catheters, and
humidifiers in hospitals.
● Cultures typically produce a greenish pigment that fluoresces
under ultraviolet light, as well as a distinctive odor.
● Reservoirs for P. aeruginosa include plants, soil, and tap water
● It can cause mild illness in healthy people and severe
infections in
people with weak immune systems.
● The leading cause of nosocomial respiratory tract infections

Talaro K.P., Chess B. 2012. Foundation in Microbiology. Ed 8. Pasadena. Mc Graw Hill.

Brooks G.L et al. Jawetz, Melnick, & Adelberg’s Medical Microbiology. Ed 27. New York. 2016. Lange.
 P. aeruginosa - Morphology

● A straight or slightly curved, Gram-negative bacillus,

arranged singly, in pairs, or in short chains.
● It is motile  a polar flagellum (may possess two or
three polar flagella)
● Pseudomonas spp. is non capsulated.
● Many strains appear mucoid by production of an
abundant of extracellular polysaccharide composed of
alginate polymers. This slime layer forms a loose
capsule or glycocalyx around the bacillus.
● Pseudomonas spp. is non sporing and fimbriated
Cornelissen C.N., Fisher B.D., Harvey R.A. 2001. Lippincott’s Illustrated Reviews: Microbiology. Ed 3. Philadelphia. Lippincott.
Talaro K.P., Chess B. 2012. Foundation in Microbiology. Ed 8. Pasadena. Mc Graw Hill.
Brooks G.L et al. Jawetz, Melnick, & Adelberg’s Medical Microbiology. Ed 27. New York. 2016. Lange.
 Virulence factor
● Virulence factors include

○ Flagella

○ Pili that aid in attachment of the bacterium to host cells.

○ A phagocytosis-resistant slime layer that contributes to the formation of biofilms.

○ Enzymes that degrade host tissues, and exotoxins (exotoxin A) that damage or destroy neutrophils and lymphocytes.

○ The LPS layer of the outer membranes also causes endotoxic shock.
● P aeruginosa produces four type III–secreted toxins that cause cell death or interfere with the host immune response to
infection.

○ Exoenzyme S and Exoenzyme T are bifunctional enzymes with GTPase and ADP-ribosyl transferase activity

○ Exoenzyme U is a phospholipase

○ Exoenzyme Y is an adenylyl cyclase

Talaro K.P., Chess B. 2012. Foundation in Microbiology. Ed 8. Pasadena. Mc Graw Hill.

Brooks G.L et al. Jawetz, Melnick, & Adelberg’s Medical Microbiology. Ed 27. New York. 2016. Lange.
 Clinical manifestation
● P. aeruginosa is a prime example of an opportunistic pathogen, with the most
serious infections seen in patients with severe burns, neoplastic disease, or
cystic fibrosis.

○ In many cases leading to endocarditis, bronchopneumonia or meningitis

(If penetrate respi track).

○ Might cause necrotizing pneumonia in CF

● Hemorrhagic necrosis of skin occurs often in sepsis caused by P aeruginosa;
the lesions, called ecthyma gangrenosum, are surrounded by erythema and
often do not contain pus.
● Otherwise-healthy persons may acquire skin rashes, wound with blue-green
pus, urinary tract infections, or outer ear infections
● Often connected to the use of community hot tubs and swimming
pools (related to the temperature).

Talaro K.P., Chess B. 2012. Foundation in Microbiology. Ed 8. Pasadena. Mc Graw Hill.

Brooks G.L et al. Jawetz, Melnick, & Adelberg’s Medical Microbiology. Ed 27. New York. 2016. Lange.
P. aeruginosa -
Pathogenesis
P. aeruginosa – Pathogenesis of Biofilm
 P. aeruginosa - Clinical diseases

● Ecthyma gangrenosum
● Wound infections
● Pulmonary disease, especially among individuals with CF
● Nosocomial urinary tract infections (UTIs)
● Endocarditis
● Bone infections
● Eye infections (keratitis, ulcers, and endophthalmitis)
● Infections following burns or trauma
● Central nervous system infections (meningitis in rare cases)
 Pseudomonas aeruginosa - Culture
● Strictly aerobic bacteria, grows over a wide range of temperatures (5–32°C), the optimum
temperature being 37°C.
● Sheep blood agar (SBA): β-hemolytic and will produce flat spreading colonies with a
characteristic metallic sheen.
● Nutrient agar (incubation for 24 hours at 37°C): large (2–3 mm in diameter), opaque,
translucent, and irregularly round colonies, characteristic musty to fruity (production of
aminoacetophenone from the amino acid tryptophan), hemolytic colonies on blood agar.
● MacConkey agar: colorless non–lactose-fermenting colonies on MacConkey media.
● Cetrimide agar: a selective medium for culture of P. aeruginosa.
● Nutrient broth: it produces a dense turbidity with surface pellicle.

Mahon C, Lehman D. Textbook of Diagnostic Microbiology. 6th ed. St. Louis, Missouri: Elsevier Saunders; 2019.
Mahon C, Lehman D. Textbook of Diagnostic Microbiology. 6th ed. St. Louis, Missouri: Elsevier Saunders; 2019.
 P. aeruginosa - Different types of pigments

● Pyocyanin: blue water-soluble

● Pyoverdin, or fluorescein: a yellow-green or yellow-brown pigment, water soluble and fluoresces under
short-wavelength ultraviolet light.
● Pyorubin is a red pigment, which is soluble in water but insoluble in alcohol.
● Pyomelanin is a brown pigment.

Mahon C, Lehman D. Textbook of Diagnostic Microbiology. 6th ed. St. Louis, Missouri: Elsevier Saunders; 2019.
Mahon C, Lehman D. Textbook of Diagnostic Microbiology. 6th ed. St. Louis, Missouri: Elsevier Saunders; 2019.
P. aeruginosa - Laboratory Diagnosis
 Treatment
● The metabolic versatility of Pseudomonas makes controlling the bacterium difficult.
● Traditionally, significant infections have not been treated with single-drug therapy because the
success rate is low and the bacteria can rapidly develop resistance when single drugs are used.

○ An extended spectrum penicillin such as piperacillin is used in combination with an

aminoglycoside, usually tobramycin.

○ Aztreonam; carbapenems such as imipenem or meropenem and fluoroquinolones, including

ciprofloxacin.

○ Effective drugs include third- and fourth-generation cephalosporins (antipseudomonas)

■ Ceftazidime, cefoperazone, and cefepime are active against P aeruginosa;

ceftazidime is often used with an aminoglycoside in primary therapy of P aeruginosa
infections, especially in patients with neutropenia.
● Multidrug resistance has become a major issue in the management of HAIs due to

○ Acquisition of chromosomal β-lactamases, extended-spectrum β-lactamases, porin channel

mutations, and efflux pumps.
Talaro K.P., Chess B. 2012. Foundation in Microbiology. Ed 8. Pasadena. Mc Graw Hill.
Brooks G.L et al. Jawetz, Melnick, & Adelberg’s Medical Microbiology. Ed 27. New York. 2016. Lange.
 P. aeruginosa - Prevention and Control

● Prevention of cross-infection of patients by attending medical and paramedical personnel.

● Prevention of contamination of equipment (dialysis machine and respiratory instruments)
● Restricting the use of broad-spectrum antibiotics.
● Pseudomonas vaccines in specialized situation (cystic fibrosis that are highly susceptible to
Pseudomonas infections)
 P. fluorescens, P. putida, or P. stutzeri

● No known virulence factors have been associated with P.

fluorescens, P. putida, or P. stutzeri.
● When infections caused by these organisms occur, they usually
involve a compromised patient exposed to contaminated medical
materials.
● Such exposure has been known to result in infections of the
respiratory and urinary tracts and wounds and bacteremia
● However, because of their low virulence, whenever these species are
encountered in clinical specimens, their significance should be
highly suspect.

Brooks G.L et al. Jawetz, Melnick, & Adelberg’s Medical Microbiology. Ed 27. New York. 2016. Lange.
Burkholderia

● Burkholderia species are also oxidase

positive non fermenters; however they
differ from Pseudomonas in being:
• Bipolar stained (safety pin
appearance)
• Resistant to polymyxin B
 Burkholderia pseudomallei

● Small, motile, oxidase-positive, aerobic Gram-negative

bacillus.
● It grows on standard bacteriologic media, forming colonies
that vary from mucoid and smooth to rough and wrinkled
and in color from cream to orange.
● It grows at 42°C and oxidizes glucose, lactose, and a
variety of other carbohydrates.
● Epizootic B pseudomallei infection occurs in sheep, goats,
swine, horses, and other animals, although animals do not
appear to be a primary reservoir for the organism.
● B pseudomallei is considered a Category B agent of
bioterrorism

Talaro K.P., Chess B. 2012. Foundation in Microbiology. Ed 8. Pasadena. Mc Graw Hill.

Brooks G.L et al. Jawetz, Melnick, & Adelberg’s Medical Microbiology. Ed 27. New York. 2016. Lange.
 Clinical menifestation
● B pseudomallei causes melioidosis (or Whitmore’s disease), which occurs
predominantly in Southeast Asia and northern Australia.
● Human infection probably originates from these sources by contamination of skin
abrasions and possibly by ingestion or inhalation.

○ Patient with diabetes mellitus is known to have higher risk of this infection.
● Melioidosis may manifest as acute, subacute, or chronic infection. The incubation
period can be as short as 2–3 days, but latent periods of months to years also
occur.
● The signs and symptoms depend upon the major sites of involvement.
● The most common form of melioidosis is pulmonary infection (atypical
pneumonia)

○ Primary pneumonitis (B pseudomallei transmitted through the upper airway

or nasopharynx)

○ Subsequent to a localized suppurative infection and bacteremia.

Talaro K.P., Chess B. 2012. Foundation in Microbiology. Ed 8. Pasadena. Mc Graw Hill.

Brooks G.L et al. Jawetz, Melnick, & Adelberg’s Medical Microbiology. Ed 27. New York. 2016. Lange.
 Treatment
• Melioidosis has a high mortality rate if untreated.
• Generally susceptible to ceftazidime, imipenem, meropenem, and
amoxicillin–clavulanic acid (also ceftriaxone and cefotaxime).
• B pseudomallei are commonly resistant to penicillin, ampicillin, first-
generation and second-generation cephalosporins, and gentamicin
and tobramycin.
 The initial intensive therapy should be for a minimum of 10–14
days with ceftazidime, imipenem, or meropenem.
 Eradication therapy with sulfamethoxazole– trimethoprim or
doxycycline should follow the intensive initial therapy and be
continued for a minimum of 3 months.

Brooks G.L et al. Jawetz, Melnick, & Adelberg’s Medical Microbiology. Ed 27. New York. 2016. Lange.
 Burkholderia cepacia complex

● Burkholderia cepacia and 17 other genomospecies comprise the B

cepacia complex.
● Grows on most media used in culturing specimens for Gram-negative
bacteria.

○ Selective media containing colistin (eg, B cepacia selective agar) is

recommended when culturing the sputum of CF patients.

○ It grows more slowly than enteric Gram-negative rods, and it may take 3
days before colonies are visible.
● Oxidase positive and lysine decarboxylase positive and produce acid
from glucose, but differentiating B cepacia from other pseudomonads,
including Stenotrophomonas maltophilia, it requires a battery of
biochemical tests.

Brooks G.L et al. Jawetz, Melnick, & Adelberg’s Medical Microbiology. Ed

27. New York. 2016. Lange.
 ● Environmental organisms able to grow in water, soil, plants, animals, and decaying vegetable
materials.
● In hospitals, members of the B cepacia complex have been isolated from a variety of
water and environmental sources from which they can be transmitted to patients HAIs
● May have asymptomatic carriage, progressive deterioration over a period of months, or rapidly
progressive deterioration with necrotizing pneumonia and bacteremia.
● People with CF and those patients with chronic granulomatous disease are particularly
vulnerable.
● B cepacia complex isolates recovered from CF patients often are multidrug resistant.
● Trimethoprim– sulfamethoxazole, meropenem, and ciprofloxacin, or alternatively
minocycline, are effective treatments.

Brooks G.L et al. Jawetz, Melnick, & Adelberg’s Medical Microbiology. Ed 27. New York. 2016. Lange.
 Antimicrobial
therapy and
susceptibility
testing of
Pseudomonad

Tille P.M. 2017. Bailey & Scott’s Diagnostic Microbiology: Bacterial

Genetics, Metabolism, and Structure. Ed 14. Missouri. Elsevier.
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