ALCANTARA, S.

▪ Jacuzzi/hot tub syndrome

→ Shanghai Fever
→ Hot tub folliculitis - infection of the hair
follicles
▪ Bacterimia leading to ecthyma gangrenosum
Chapter 21: Nonfermenting Gram-Negative → Ecthyma gangrenosum is characterized as
Bacilli a burn/deep wound infections; forms
blisters that leads to tissue necrosis
Chapter Outline:
II. Virulence Factor
I. General Characteristics of Nonfermenters
→ Clinical Infections 1. Protease, hemolysins, lecithinase,
→ Biochemical Characteristics and elastase and DNase - Enzymes that
Identification degrades certain molecules
2. Exotoxins:
II. Clinically significant Nonfermentative, Gram-
• Endotoxin, motility, pili and capsule
Negative Bacilli
→ Pseudomonas aeruginosa
• Exotoxin A (inhibits protein
→ Acinetobacter synthesis)
→ Stenotrophomonas 3. Alginate (Polysaccharide polymer in
→ Burkholderia mucoid strains)
4. Inherently resistance to a number of
microbial agents
I. General Characteristics
III. Identifying Characteristics
→ Grow in MacConkey as colorless colonies
Gram-negative obligate aerobe -that uses
Fail to acidify O-F Media, overlaid with
oxygen for metabolism through cellular
mineral oil
respiration; it is encapsulated and does not
→ Fail to acidify TSI (Triple Sugar Iron)
ferment lactose. It makes spores. It is catalase (+),
→ Most isolates in Oxidase Positive citrate and oxidase (+)
→ Resistance to a variety of classes
of antimicrobial agents
Abundant in the environment and has flagellum
at one pole surrounded by pili.

II. Clinical Significant Species 1. Strict aerobic

2. Pigmented
A. Pseudomonas aeruginosa 3. Fruity, Grape-like Odor (2-
B. Acinetobacter aminoacetophenone)
C. Stenotrophomonas 4. Growth at 42C
5. Grows in Cetrimide Agar
6. Acetamide Positive
Pseudomonas aeruginosa
▪ Pigments:
I. Clinical Significance → Fluirescien (Pyoverdin) - Yellow
→ Pyocyanin - Blue
1. Gram-negative obligate aerobe - It uses → Pyorubin - Red
oxygen for metabolism → Pyomelanin - Brown/Black
2. Most commonly isolated species
▪ 75% of nonfermenters in nosocomial Pyocyanin pigment - Sweet grapelike fruit
bacteremias and odor with oxidative damage to the cell
▪ 5% to 15% of nosocomial infections
3. Causes bacteremia, wound infection,
pulmonary disease (CF patients), UTI,
endocarditis, and meningitis
4. Otitis media and Jacuzzi/Hot tub
syndrome
 ALCANTARA, S.Y

Pyoverdin pigment - yellow-green color; iron II. Identifying Characteristics

chelator that binds iron and transport it to P. 1. A. baumanii
aeruginosa ▪ Saccharolytic (glucose oxidizing),
nonhemolytic strains
2. A. iwoffi
▪ Asaccharolytic (cannot oxidice glucose),
nonhemolytic strains
3. A. haemolyticus
▪ -hemolytic strains (complete hemolysis of
RBC)

 Plump, paired gram negative coccobacilli

IV. Clinical Infections  Nonfermenters & commonly found in soil
and water
1. Infect damage tissues and infect those
with weakened immune system
2. An opportunistic nosocomial pathogen of
immunocompromised indivuduals, it can
infect:
▪ Pulmunary tract
▪ Urinary tract
▪ Burns/Wounds
▪ Causes other blood infections
3. If colonization occurs in critical body organ  A. baumanni may exhibit purplish hue
(lungs, urinary tract or kidneys) the results (due to lactose oxidation)
can be fatal  A. buamanni colony appears with a blue-
4. Cystic fibrosis patients appear to be grey (cornflower blue) center in EMB
predisposed to the infection of the lungs to
P. aeroginosa
5. Can be found in medical equipments Stenotrophomonas spp.
(catheters) → infections in hospital/clinics
and post surgical infections Stenotrophomonas maltophilia
6. Can cause community-acquired
pneumonias and associated with Hot tub I. Clinical Significance
rash
1. Contaminants in blood drawing
equipment (collection tubes, disinfectants,
Acinetobacter spp. transducers, venous catheters, etc.)
2. Opportunistic pathogen (3rd most
commonly isolated among nonfermenters)
I. Clinical Significance 3. Intrinsic resistance commonly used
antibacterial agent (aminoglycosides and
1. Found commonly in soil and water beta lactams)
2. Associated with ventilators, humidifiers,
catheters, etc. II. Identifying Characteristics
3. Opportunistic pathogen (1-3% of all
nosocomial infection, 2nd most commonly 1. Lavender green pigment in BAP
isolated nonfermenter gram-negative 2. Ammonia-like smell
bacilli 3. Oxidizes glucose: (+)
4. Outbreaks in Acinetobacter infection 4. Oxidizes maltose: (+)
typically accord in ICU of healthcare
facility that causes variety of diseases:
▪ Pneumonia
▪ Serious Blood and wound Infections
 ALCANTARA, S.Y

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

Burkholderia II. Identifying Characteristics

▪ B. cepacia 1. Bipolar staining (safety pin) in gram stain

▪ B. pseudomallei 2. Gram negative rods; motile; swell dwelling
▪ B. gladioli bacterium
▪ B. mallei 3. Smooth to wrinkled colonies in BAP
4. Wrinkled and deep pink in Ashdown
media
5. "Earthy odor"
Burkholderia cepacia

I. Clinical Significance

1. Most clinically significant Burkholderia

specie
2. Gram-negative, nonfermenting bacilli
3. Found in various aquatic environments
4. Low virulence and usually colonizers of
fluid found in hospital Culture:
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 Burkholderia glandoili

by lung congestion and infection and
malabsorption of nutrients by the pancreas.
▪ Associated with patients with CF and CDG
II. Identifying Characteristics (Cystic fibrosis and Chronic Granulomatous
Disease)
1. Smooth and slightly raised; dirtlike odor in
BAP
Burkholderia glandoili
2. NLF; become dark pink to red after 4-7
days on MAC
3. Most strains are ONPG positive ▪ Glanders disease - disseminated or FARCY
4. Produces a nonfluorescing yellow/green zoonotic infection (Donkey)
pigment ▪ Nonmotile
 ALCANTARA, S.Y

III. Laboratory Diagnosis ▪ Used on the differentiation among

Enteribacteriaciaea on the basis of their
1. Nitrate reduction sugar fermentation pattern
2. Triple Sugar Iron Agar (TSI)
3. Acetamide Utilization
4. Growth at 42C
5.

1. Nitrate Reduction

▪ Test that determines the production of

enzyme called nitrate nitrate reductase
▪ It result in reduction of nitrate
▪ Bacterial species may be differentiated on
the basis 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 Acid to form a
complex. which then reacts with the Alpha
naphythylamine gives red presipitate
(water-soluble azo dye). Expected Results:
▪ Positive result: Cherry red coloration on
addition of reagent A and B; Absence of 1. Tube 1
red color development upon addition of ▪ Reaction: BK/K H2S- (Alkaline/Alkaline)
zinc powder ▪ It means an abscense of carbohydrate
▪ Negative result: Development of a red color fermentation results
on addition of the zinc powder ▪ Organism: Pseudomonas

2. Tube 2
▪ Reaction: K/A H2S+ (Alkaline/Acid)
▪ Indicative of dextrose fermentation only
▪ Organism: Serratia Providencia Shigella

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

2. Triple Sugar Iron Agar (TSI) 4. Tube 4

▪ Reaction: A/A H2S- w/ gas (Acid/Acid)
▪ Microbiological test named for its ability to
test microorganisms ability to ferment sugars 5. Tube 5
and produce hydrogen sulfide ▪ Reaction: A/A H2S+ w/ gas
▪ Agar slant of special medium with a multiple ▪ Cause blackening and gas or cracks
sugar constituting a pH sensitive dye underneath the tube
(Phenol red w/ 1% lactose, 1% sucrose and
0.1% glucose, sodium thiosulfate, ferrous
sulfate)
▪ TSI test used for the differentiation of the
members of Enterobacteriacaea family
from the other gram-negative rods
 ALCANTARA, S.Y

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).
▪ The shift in pH turns the bromothymol blue
(pH indicator) and the medium rom the
green → blue = POSITIVE TEST

Positive Results: Determination of

acetamide resulting in a blue color
→ Pseudomonas aeroginosa

Negative results: No color change

→ Stenotrophomonas maltophia
→ Pseudomonas fluorescens
→ Pseudomonas putida

4. Growth at 42C

▪ Test the ability of an organism to grow at

42C

Positive: Good grow that both 35C

and 42C
→ Pseudomonas aeroginosa
→ Stenotrophomonas maltophilia

Negative: No growth at 42 but good

growth at 35C
→ Pseudomonas fluorescens
→ Pseudomonas putida

Biochemical Physiologic Characteristics

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