ALCANTARA, S.

 Ecthyma gangrenosum is characterized as a

burn/deep wound infections; forms blisters that
leads to tissue necrosis
Chapter Outline: II. Virulence Factor
Chapter 21: Nonfermenting Gram-Negative Bacilli
I. General Characteristics of Nonfermenters
 Clinical Infections 1. Protease, hemolysins, lecithinase, elastase and
 Biochemical Characteristics and Identification DNase - Enzymes that degrades certain molecules
2. Exotoxins:
II. Clinically significant Nonfermentative, Gram-Negative  Endotoxin, motility, pili and capsule
Bacilli  Exotoxin A  (inhibits protein synthesis)
 Pseudomonas aeruginosa 3. Alginate (Polysaccharide polymer in mucoid
 Acinetobacter strains)
 Stenotrophomonas 4. Inherently resistance to a number of microbial
 Burkholderia
I. General Characteristics agents

III. Identifying Characteristics

Gram-negative obligate aerobe -that uses oxygen for

metabolism through cellular respiration; it is encapsulated
and does not ferment lactose. It makes spores. It is catalase
(+), citrate and oxidase (+)

Abundant in the environment and has flagellum at one pole

 Grow in MacConkey as colorless coloniesFail to surrounded by pili.
acidify O-F Media, overlaid with mineral oil
 Fail to acidify TSI (Triple Sugar Iron) 1. Strict aerobic
 Most isolates in Oxidase Positive 2. Pigmented
3. Fruity, Grape-like Odor (2-aminoacetophenone)
 Resistance to a variety of classes of antimicrobial
4. Growth at 42C
II. Clinical Significant Species 5. Grows in Cetrimide Agar
6. Acetamide Positive
agents
 Pigments:
 Fluirescien (Pyoverdin) - Yellow
A. Pseudomonas aeruginosa  Pyocyanin - Blue
B. Acinetobacter  Pyorubin - Red
C. Stenotrophomonas  Pyomelanin - Brown/Black

Pseudomonas aeruginosa Pyocyanin pigment - Sweet grapelike fruit odor with

oxidative damage to the cell
I. Clinical Significance

1. Gram-negative obligate aerobe - It uses oxygen for

metabolism
2. Most commonly isolated species 
 75% of nonfermenters in nosocomial
bacteremias and
 5% to 15% of nosocomial infections Pyoverdin pigment - yellow-green color; iron chelator
3. Causes bacteremia, wound infection, pulmonary that binds iron and transport it to P. aeruginosa
disease (CF patients), UTI, endocarditis, and
meningitis
4. Otitis media and Jacuzzi/Hot tub syndrome 
 Jacuzzi/hot tub syndrome
 Shanghai Fever
 Hot tub folliculitis - infection of the hair follicles
 Bacterimia leading to ecthyma gangrenosum
 ALCANTARA, S.Y

IV. Clinical Infections

 A. baumanni may exhibit purplish hue (due to
1. Infect damage tissues and infect those with lactose oxidation)
weakened immune system  A. buamanni colony appears with a blue-grey
2. An opportunistic nosocomial pathogen of (cornflower blue) center in EMB
immunocompromised indivuduals, it can infect:
 Pulmunary tract
 Urinary tract Stenotrophomonas spp.
 Burns/Wounds
 Causes other blood infections Stenotrophomonas maltophilia
3. If colonization occurs in critical body organ (lungs,
urinary tract or kidneys) the results can be fatal I. Clinical Significance
4. Cystic fibrosis patients appear to be predisposed to
the infection of the lungs to P. aeroginosa 1. Contaminants in blood drawing equipment
5. Can be found in medical equipments (catheters)  (collection tubes, disinfectants, transducers, venous
infections in hospital/clinics and post surgical catheters, etc.)
infections 2. Opportunistic pathogen (3rd most commonly
6. Can cause community-acquired pneumonias and isolated among nonfermenters)
associated with Hot tub rash 3. Intrinsic resistance commonly used antibacterial
agent (aminoglycosides and beta lactams)
Acinetobacter spp. II. Identifying Characteristics

1. Lavender green pigment in BAP

I. Clinical Significance 2. Ammonia-like smell
3. Oxidizes glucose: (+)
1. Found commonly in soil and water 4. Oxidizes maltose: (+)
2. Associated with ventilators, humidifiers, catheters,
etc.
3. Opportunistic pathogen (1-3% of all nosocomial
infection, 2nd most commonly isolated
nonfermenter gram-negative bacilli
4. Outbreaks in Acinetobacter infection typically
accord in ICU of healthcare facility that causes
variety of diseases:
 Pneumonia
 Serious Blood and wound Infections

II. Identifying Characteristics

1. A. baumanii
 Saccharolytic (glucose oxidizing), nonhemolytic
strains
2. A. iwoffi
 Asaccharolytic (cannot oxidice glucose),
nonhemolytic strains
3. A. haemolyticus Burkholderia
 -hemolytic strains (complete hemolysis of RBC)
 B. cepacia
 Plump, paired gram negative coccobacilli  B. pseudomallei
 Nonfermenters & commonly found in soil and water  B. gladioli
 B. mallei

Burkholderia cepacia

I. Clinical Significance
 ALCANTARA, S.Y

1. Most clinically significant Burkholderia specie

2. Gram-negative, nonfermenting bacilli  Associated with patients with CF and CDG (Cystic
3. Found in various aquatic environments fibrosis and Chronic Granulomatous Disease)
4. Low virulence and usually colonizers of fluid found
in hospital
Burkholderia glandoili
5. Isolated from irrigation, anesthetics, nebulizers,
detergents and disinfectants
6. Pneumonia in patients with cystic fibrosis or
chronic granulomatous disease
7. Onion bulb rot in plants and foot rot in humans
8. Intrinsic resistance to antibiotics
9. Rarely causes infection in healthy host

Cystic Fibrosis - hereditary disorder characterized by lung  GlandersBurkholderia

disease - disseminated
glandoili or FARCY
congestion and infection and malabsorption of nutrients by zoonotic infection (Donkey)
the pancreas.  Nonmotile

II. Identifying Characteristics

III. Laboratory Diagnosis
1. Smooth and slightly raised; dirtlike odor in BAP
2. NLF; become dark pink to red after 4-7 days on
MAC 1. Nitrate reduction
3. Most strains are ONPG positive 2. Triple Sugar Iron Agar (TSI)
4. Produces a nonfluorescing yellow/green pigment 3. Acetamide Utilization
4. G r o w t h a
Burkholderia pseudomallei

I. Clinical Significance

1. Causative agent of "Melioidosis" (pulmonary

infection)
2. Meliodosis - White morse disease or "Vietnam
Time Bomb (present in body for years)
3. Potential for bioterrorism weapon C
5.

II. Identifying Characteristics 1. Nitrate Reduction

1. Bipolar staining (safety pin) in gram stain  Test that determines the production of enzyme called
2. Gram negative rods; motile; swell dwelling nitrate nitrate reductase
bacterium  It result in reduction of nitrate
3. Smooth to wrinkled colonies in BAP  Bacterial species may be differentiated on the basis
4. Wrinkled and deep pink in Ashdown media
5. "Earthy odor"

of their ability to reduce nitrate to nitrate or

nitrogenous gases
 Base on the detection of nitrate and its ability to form
red coumpound when it reacts with the Sulfanilic
Culture: Acid to form a complex. which then reacts with the
Alpha naphythylamine gives red presipitate (water-
soluble azo dye).
 ALCANTARA, S.Y

 Positive result: Cherry red coloration on addition of  Reaction: A/A H2S+ w/ gas
reagent A and B; Absence of red color development  Cause blackening and gas or cracks underneath
upon addition of zinc powder the tube
 Negative result: Development of a red color on
addition of the zinc powder

2.

3. Acetamide Utilization

 Determine the ability of an organism to use

acetamide as the sole source of carbon
 Indicator: Bromthymol blue (acetamide 
ammonia)
 Growth is indicative of a positive test for acetamide
utilization, so when bacteria metabolizes acetamide
by the enzymatic action upon acetylamidase , the
ammonium salts are broken down into ammonia
(increases alkalinity).
Triple Sugar Iron Agar (TSI)  The shift in pH turns the bromothymol blue (pH
indicator) and the medium rom the green  blue =
Microbiological test named for its ability to test POSITIVE TEST
microorganisms ability to ferment sugars and
produce hydrogen sulfide Positive Results: Determination of acetamide
 Agar slant of special medium with a multiple sugar resulting in a blue color
constituting a pH sensitive dye (Phenol red w/ 1%  Pseudomonas aeroginosa
lactose, 1% sucrose and 0.1% glucose, sodium
thiosulfate, ferrous sulfate) Negative results: No color change
 TSI test used for the differentiation of the members  Stenotrophomonas maltophia
of Enterobacteriacaea family from the other gram-  Pseudomonas fluorescens
negative rods  Pseudomonas putida
 Used on the differentiation among
Enteribacteriaciaea on the basis of their sugar 4. Growth at 42C
fermentation pattern
Expected Results:
 Test the ability of an organism to grow at 42C
1. Tube 1
Positive: Good grow that both 35C and
 Reaction: BK/K H2S- (Alkaline/Alkaline)
 It means an abscense of carbohydrate 42C
fermentation results  Pseudomonas aeroginosa
 Organism: Pseudomonas  Stenotrophomonas maltophilia

2. Tube 2 Negative: No growth at 42 but good growth

 Reaction: K/A H2S+ (Alkaline/Acid) at 35C
 Indicative of dextrose fermentation only  Pseudomonas fluorescens
 Organism: Serratia Providencia Shigella  Pseudomonas putida

3. Tube 3
 Reaction: K/A H2s+ w/gas Biochemical Physiologic Characteristics
 Gas production, bubble/cracks agar is an
indicative of the production of gas formation or
Carbon dioxide and Hydrogen gases
 Organism: Citrobacter, Salmonella Proteus

4. Tube 4
 Reaction: A/A H2S- w/ gas (Acid/Acid)

5. Tube 5
 ALCANTARA, S.Y

PAG UMABOT KA HANGGANG DITO,

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