Chapter 20

The Gram-Negative
Bacilli of Medical
Importance

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Aerobic Gram-Negative Nonenteric Bacilli
• Large, diverse group of
non-spore-forming
bacteria
• Wide range of habitats –
large intestines (enteric),
zoonotic, respiratory, soil,
water
• Most are not medically
important; some are true
pathogens, some are
opportunists
• All have outer membrane
lipopolysaccharide of the
cell wall – endotoxin *
 Aerobic Gram-Negative
Nonenteric Bacilli
• Pseudomonas and Burkholderia –
opportunistic pathogens

• Brucella and Francisella – zoonotic

pathogens

• Bordetella and Legionella – mainly human

pathogens

*
Pseudomonas: The Pseudomonads
• Small gram-negative rods with a single polar flagellum

• Free living
– Primarily in soil, sea water, and fresh water; also
colonize plants and animals

• Important decomposers and bioremediators

• Frequent contaminants in homes and clinical settings

• Use aerobic respiration; do not ferment carbohydrates

• Produce oxidase and catalase

• Many produce water soluble pigments *

 Pseudomonas aeruginosa
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• Common inhabitant of soil

and water
• Intestinal resident in 10%
normal people
• Resistant to soaps, dyes,
quaternary ammonium © Science VU/Visuals Unlimited

disinfectants, drugs, drying

• Frequent contaminant of
ventilators, IV solutions,
anesthesia equipment
• Opportunistic pathogen
© Bottone E.J. and Perez A.A., II, The Mount Sinai Hospital New York

*
 Pseudomonas aeruginosa
• Common cause of nosocomial infections in hosts with
burns, neoplastic disease, cystic fibrosis
• Complications include pneumonia, UTI, abscesses,
otitis, and corneal disease
• Endocarditis, meningitis, bronchopneumonia
• Grapelike odor and greenish-blue pigment (pyocyanin)
• Multidrug resistant
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*
© Kathy Park Talaro © Kathy Park Talaro
 Related Gram-Negative Aerobic Rods
• Genera Burkholderia, Acinetobacter,
Stenotrophomonas

• Similar to pseudomonads

• Wide variety of habitats in soil, water, and related

environments

• Obligate aerobes; do not ferment sugars

• Motile, oxidase positive

• Opportunistic
*
 Burkholderia
• Burkholderia cepacia – active in biodegradation of a
variety of substances; opportunistic agent in respiratory
tract, urinary tract, and occasionally skin infections; drug
resistant
• B. pseudomallei – generally acquired through penetrating
injury or inhalation from environmental reservoir; wound
infections, bronchitis and pneumonia, septicemia
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Dense zone
with infection

From Kongsaengdao S, et al, “Treatment of Survivors after the Tsunami,” New England Journal of Medicine Vol.
*
352:2654-2655, © 2005 Massachusetts Medical Society. All rights reserve.
From Kongsaengdao S, et al, “Treatment of Survivors after the Tsunami,” New England Journal of
Medicine, Vol. 352:2654-2655, © 2005 Massachusetts Medical Society. All rights reserved
(a (b
 Acinetobacter and
Stenotrophomonas
• Acinetobacter baumanii – nosocomial and
community acquired infections; wounds,
lungs, urinary tract, burns, blood; extremely
resistant – treatment with combination
antimicrobials

• Stenotrophomonas maltophilia – forms

biofilms; contaminant of disinfectants dialysis
equipment, respiratory equipment, water
dispensers, and catheters; clinical isolate in
respiratory soft tissue, blood, CSF; high
resistance to multidrugs
*
 Concept Check:
The main reservoir for Pseudomonas
aeruginosa is

A. Soil and Water

B. Infected Individuals
C. Asymptomatic Carriers
D. Wild Animals
 Brucella and Brucellosis
• Tiny gram-negative coccobacilli
• 2 species:
– Brucella abortus (cattle)
– Brucella suis (pigs)

• Brucellosis, malta fever, undulant fever, and Bang

disease – a zoonosis transmitted to humans from
infected animals
• Fluctuating pattern of fever – weeks to a year

• Combination of tetracycline and rifampin or

streptomycin
• Animal vaccine available
• Potential bioweapon *
Agglutination Titer Test for Brucellosis
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Antibody( Antibody( 1 2 3 4 5 6
+) –)
for for
1:10
Brucella Brucella
Antige
n 1:20
1:40
1:80
1:160
Brucella
POSITIVE NEGATIVE
cells 1:320
REACTION REACTION
1:640
1:1280

Picture courtesy of Vircell

(a) Serum (b) Serum (c) The numbered lanes are

containing lacking individual serum samples.
antibodies specific specific Brucella The serum titer can be
to antibodies does determined
Brucella captures not by diluting it through wells A
to H, *
the react, and
test cells (antigen) antigens then looking for the last well
Francisella tularensis and Tularemia
• Causes tularemia, a zoonotic disease of mammals
endemic to the northern hemisphere, particularly
rabbits
• Transmitted by contact with infected animals, water
and dust or bites by vectors
• Headache, backache, fever, chills, malaise, and
weakness
• 10% death rate in systemic and pulmonic forms
• Intracellular persistence can lead to relapse
• Gentamicin or tetracycline
• Attenuated vaccine
• Potential bioterrorism agent
*
 Bordetella pertussis
• Minute, encapsulated coccobacillus
• Causes pertussis or whooping cough, a communicable
childhood affliction
• Acute respiratory syndrome
• Often severe, life-threatening complications in babies
• Reservoir – apparently healthy carriers
• Transmission by direct contact or inhalation of aerosols
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*
CDC
 Bordetella pertussis
• Virulence factors Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

– Receptors that 5.
0
4.
recognize and bind to 5
4.
ciliated respiratory

Reported Cases per 100,000

0
3.
epithelial cells 5
3.
0
– Toxins that destroy and 2.
5
dislodge ciliated cells 2.
0

Population
1.
• Loss of ciliary mechanism 5
1.
leads to buildup of mucus 0
0.

and blockage of the 5

0.
0197 198 198 199 199 200 200 201
airways 6 1 6 1Yea
r
6 1 6 0

• Vaccine – DTaP –
acellular vaccine contains
toxoid and other Ags
*
 Alcaligenes
• Live primarily in soil and water

• May become normal flora

• A. faecalis – most common clinical species

– Isolated from feces, sputum, and urine
– Occasionally associated with opportunistic
infections – pneumonia, septicemia, and
meningitis

*
 Legionella pneumophila and
Legionellosis
• Widely distributed in water
• Live in close association with amoebas
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Legionella
bacteria

CDC CDC
*
(a (b Amoeba
) ) cell
 Legionella pneumophila and
Legionellosis
• 1976 epidemic of pneumonia afflicted 200
American Legion members attending a
convention in Philadelphia and killed 29
• Legionnaires disease and Pontiac fever
• Prevalent in males over 50
• Nosocomial disease in elderly patients
• Fever, cough, diarrhea, abdominal pain,
pneumonia fatality rate of 3-30%
• Azithromycin is main treatment *
 Enterobacteriaceae Family
• Enterics

• Large family of small, non-spore-forming gram-

negative rods

• Many members inhabit soil, water, decaying matter,

and are common occupants of large bowel of animals
including humans

• Most frequent cause of diarrhea through enterotoxins

• Enterics, along with Pseudomonas sp., account for

almost 50% of nosocomial infections
*
 Enterobacteriaceae Family
• Facultative anaerobes, Copyright © The McGraw-Hill Companies, Inc. Permission required for reproduction or display.

grow best in air Lactose

(+)
Lactose
(–)

• All ferment glucose, Lactose

(–)
reduce nitrates to nitrites, Lactose
(+)
H2S (+) Lactose
(–)
oxidase negative, and
catalase positive
• Divided into coliforms
(lactose fermenters) and
non-coliforms (non-
lactose fermenters)
• Enrichment, selective and
differential media utilized
for screening samples for
© Kathy Park Talaro © Kathy Park Talaro
(a (b
) )
pathogens
*
 The Enteric Genera
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Colonies from selective media inoculated into triple-sugar

iron (TSI)
Rapid lactose fermentation on
+ Lactos (TSI) Lactos –
e
+ Glucos e
Glucos +
e e
Motilit Phenylalanine
+ y – + (PA) –

Indol Voges- H2S Citrat

+ e – +Proskauer
(VP – + – + e –
)
Klebsiell Moellerell Proteu
H2S a a s
+ – Citrat Motilit
+ e – + y –

Providenci Morganell ONPG

Erwinia – VP+Enterobact a a + –
VP-
Citrobacter er
– Hafni Edwardsiell
Citrat Citrobacter a a Ureas
+ e – + e –

Escherichi See table 20.2 for a brief Yersini Shigell

LD ONPG
discussion
+ C –a gelatinas a a
of each differential test + e –

Kluyver Citrobact Serrati Salmonell

a er a a *
 Colors
Reacted
d Colors
G

Gas
Uninoculate
G lu
lu G co
c pr as se

production
Ly ose oLdy
desin desuicn
e ca e e cateio
O rb O rb n
e d rni ox e d rni ox
e ecathin yla e ecathin yla
H rb s s
2S ox H rbo
In 2S xy

Indole
yl do / la
as s
Ad Adle
l on l on
i to i to
La La
ct ct
os os
Ar e Ar e
e ab e ab
in in
So os So os
l rb l rb
Vo ito Vo ito
ge ge
Pr s- Pr s-
os os
A*

ka kDa
Du
B*
Voges- PA
Proskauer
PA uelcr PA ulecr
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i to i to
U U
re l/ re l/
a a
C C
BBL Enterotube II, Rapid

B*A
*

e itra e itra
t t
Biochemical Testing of Enterics

*
 Antigenic Structures
and Virulence Factors
Complex surface antigens contribute to
pathogenicity and trigger immune response:
• H – flagellar Ag
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• K – capsule
and/or fimbrial Capsule (K antigen, or

Ag Vi in
Salmonella

• O – somatic or
cell wall Ag – all Somatic (O antigen, or
cell wall
have antigen)

• Endotoxin Flagellar (H
antigen)
• Exotoxins
*
 Concept Check:
Which of the following is true of the enterics?

A. Some are normal flora

B. Some release endotoxin
C. Some release enterotoxins
D. All of the above are true
 Escherichia coli: The Most
Prevalent Enteric Bacillus
• Most common aerobic and non-fastidious
bacterium in gut

• 150 strains

• Some have developed virulence through plasmid

transfer, others are opportunists